} I have a theory question about Critical Point Drying that has been } bothering me. I know that the specimen is placed in a "bomb", and then } transition fluid replaces the dehydrating fluid in the specimen, and the } temperature is raised to the critical point, which in turn raises the } critical pressure in the bomb, so that the specimen is in a sense } immersed in a dense vapor phase devoid of liquid air/interface, and the } vapor is slowly released until the vessel is at atmospheric pressure. } But with the drop in pressure, even though it is slow, below the } critical pressure for that transition fluid, why doesn't this } precipitate a condensation of the vapor back to a liquid???? } } Is the temperature slowly increased beyond the critical temperature, to } correspond to a new critical temp. for the lower pressure?
} For the veterans, I'm sorry to bug them with elementary questions like } this, but I can't find the answer in any book. } } Garry
Yes - the temperature of the whole system is kept sufficiently high so that as the pressure drops, it doesn't pass through the vapour/liquid transition again.
So the system is taken in a loop around the critical point, as illustrated:) This also highlights a minor problem that you may run into in some labs - if the ambient lab temperature is too high, the CO2 is always a vapour, so the bomb may need cooling to start with, just to get liquid CO2. This caused me some difficulties when installing a system in Jakarta at the beginning of the year!
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
The Dec 1997 archives are now on-line at the MSA WWW Site http://www.msa.microscopy.com.
Since I just finished writing the annual report to MSA Council on the ListServer operations here are some year-end statistics for those ListServer Subscribers that might be curious.
The Listserver has received 5403 postings this year for an average of 14.8 messages/day.
The Email archives on the MSA WWW Site now contain 29.7 Mbytes of text and have been accessed 2126 times.
The Microscopy ListServer has delivered just over 11,000,000 Email messages to 2000+ subscribers in 49 Countries while the MSA WWW Site has had 16,510 HomePage logins and delivered just over 9,800,000,000 bytes of information to WebSurfers from 95 different countries.
For you trivia buffs (or those of you that are just bored with what ever your supposed to be doing) that averages to an Email message being sent out of the server about every 350 milliseconds and a WWW byte about every 3 milliseconds during 1997.
I am interested in hearing from someone who has done cryo-electron microscopy on hydrated samples using a cold stage. I have never done this myself, and am unable to find much mention of it in the books, but was wondering what kinds of skills I would have to develop in order to do this sort of microscopy.
To all the people who answered my question about CPD, thankyou very much, it was helpful.
In order to clarify the process conditions of critical point dryer, thermodynamics analysis should be introduced. The critical point of substance needs to be descripted by three properties, e.g. temperature (T), pressure (P) and specific volume (v). The later is defined as the ratio of v=V/m, where V is the volume of chamber and m is the mass of substance in the chamber. It is easy to understand by common sense that there is no way to achieve the critical point if the amount of CO2 in the chamber is less than certain level, as most of us know, say 10% of CO2 in the chamber.
T-v diagram can be used to show the process of liquid-gas phase inter-changes as following.
Case 1: Put above certain amount of CO2 into chamber and heat it: the T and P increase, finally c.p can be achieved, the v of gas equals to the v of liquid, no interface between gas and liquid exists(point c.p). Case 2: After c.p, the gas is slowly vented, resulting m decreasing, V (chamber) is constant, thus v=V/m increases (important point here!). 1. As v increases and T is constant, the system will shift along with points of A, B and C,and goes through isobar curves meaning the pressure is decreasing. It is clear that as v increasing, the system has no way to back to liquid condition, instead, it goes into gas condition deeply. 2.As v increases and T decreases slowly(turn off heater): the system will shift along with the points of a, b, and c, which are still staying in the gas phase. 3.In an extremely condition, where the chamber is cooled very fast, say using ice-CO2 or LN2 to cool it, the T drops sharply, it is a chance for the system to back to liquid phase, but it is not in the normal operation condition. Conclusions: 1. In the normal operation condition, T does not drop sharply, the C.P.D has no chance to back to liquid phase, no matter how fast the gas is releasing. 2. The slowly ventilation of gas is required in order to prevent the sample from "pop-corn", which will damage the sample physically.
Hopeful it helps.
****************************************** Zhiyu Wang, senior technician Electron Microscope Lab Western Kentucky University(WKU) Bowling Green KY 42101
Phone: (502)745-5993(office) email: wangz-at-pulsar.cs.wku.edu ****************************************** Looking for new position +++++++++++++++++++++++++++++++++++++++++
Do you have access to a conventional ion mill??? I had a lot of success = preparing surfaces or EBSD by giving them a low angle ion mill treatment = for 15-30 minutes. This seemed to knock off any surface oxide that may = have formed and seemed to remove any small damage layer near the surface = induced by mechanical polishing. For many materials (metals included), = I skipped a chemical polish altogether and simply mechanically polished = with a final step using Syton, followed by the ion milling step.
Hope this helps,
Tom
Thomas C. Isabell, Ph.D. Research Scientist E.A. Fischione Instruments, Inc. tci-at-fischione.com webpage: www.fischione.com
-----Original Message-----
Does anyone have experience with preparation of CoPt(20%Pt) for SEM and = EBSD. Brand new material for me and aqua regia so far doesnt seem to be the = right approach.Any suggestions would be appreciated. Thanks Allen Miller
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% %%%%%%%%%%%%%%%%%%%%%%%= %%% %%%%%%%%% Patty Miller Stained Panes momiller-at-ccia.com
There is a book entitled 'Low Temperature Methods in Biological Electron Microscopy", written by A. W. Robards and U. B. Sleytr that might be of some interest to you. This was published in 1985 by Elsevier as Vol. 10 (ISBN 0-444-80684-9 pbk & ISBN 0-444-80685-7 hbk) in the series 'Practical Methods in Electron Microscopy' that is edited by Dr. Audrey M. Glauert.
The two most recent volumes in this series: No. 15, Vacuum Methods in Electron Microscopy, and No. 16, X-ray Microanalysis for Biologists, have been published by Portland Press (sales-at-portlandpress.co.uk) and are marketed in the USA by Ashgate Publishing (info-at-ashgate.com).
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Hello everyone First of all - HAPPY NEW YEAR! I am planning to stain the tubulin part the cytoskeleton in live protozoa. Microinjection with labled tubulin is not possible - the cells are rather delicate. Any suggestions of fluorochromes and technioques are welcome.
I have some cover slides made by a company named ASSISTENT. According to the box, the company is(was) located in Germany. The cover slides are very small (5 mm to 8 mm diameter) and are ideal for one of my uses. I'd like to locate this company, a replacement, or anyone else making such a product. Can you please help me?
Reply directly to me if you do not want to reply to everyone. spruance-at-infinet.com
Thank you to the many people who responded with helpful suggestions about dehydration. The basis for my query is isolation: it is hard to get anything shipped here. We are at the western edge of the Pacific Ocean (in fact, our West Coast overlooks the Philippine Sea. The humid tropical climate is an added factor.
The following suggestions were received:
W. Muss suggested Acetonitrile: 96% EtOH=1:1 to 3 x 15-20 minutes pure Acetonitrile; the medium is 100% miscible with water as well with resins, like EPON or its substitute epoxy resins. I use the Acetonitrile schedule (from EtOH 100% to AN 3x 10 min) in routine diagnostic tissue specimen processing to EPON/Epoxy resin embedding since now 12 years without problems.
A. Greene suggested Everclear: When having difficulties obtaining Absolute ETOH, I have found that "Everclear" (from a liquor store) usually works. It is only 95% but then again, as soon as you open a bottle of absolute, it ceases to be absolute, due to its hygroscopic nature.
H. Ortega: isopropanol
R. Olley suggested also Isopropanol (Propan-2-ol), and disapproved acetone: Isopropanol (or Propan-2-ol, as we are told to call it these days) in many ways behaves similarly to ethanol. It is somewhat more viscous and less volatile, but not overwhelmingly so. Moreover, it is miscible with water and should be usable for dehydrating in stages of increasing alcohol concentration.
Acteone is somewhat fiercer, might go for lipids, and when it evaporates tends to chill the specimen and pull condensation out of the air.
M. Peterson suggests acetone: I use acetone to dehydrate samples which are to be embedded in Spurr's, or Epon-Araldite. I still take care to use dry acetone, although Epon-Araldite reportedly tolerates some water. This has worked successfully for plant tissues, for years. I've seen Spurr's become very brittle, possibly as the result of incomplete dehydration.
D. Feely suggested the use of molecular sieve with with absolute EtOH to keep it dry: To insure that you use absolute ETOH for the final step, keep your 100% in a jar with dry "molecular seive" and use only what you need. MAke sure the bottle is tightly stoppered and let it sit for a few days to be sure it is dry and to let the small particals settle out.
M. Kroez says that acetone works ok for delicate tissues. Propylene-glycol is suggested as the final step.: for dehydration we use either a graded series of ethanol OR acetone. This works very well, even on delicate tissues like e.g. bone marrow. You can also use propylene-glycol as the last step of dehydration, after absolute ethanol or acetone.
P. Oshel recommends care in selecting the appropriate dehydrating agent for the medium: Acetone can be an excellent dehydrating agent. *But*, which is superior, acetone or EtOH, depends on the embedding medium and your specimen. If you're embedding in paraffin, use EtOH, as that's better for transferring to xylene/Histoclear etc., if in resin, some resins do better with acetone, some with EtOH, some don't care.
Your specimens may care a great deal. I've used EtOH for the specimens you mention with great success.
Thanks to everyone who kindly responded. I think I am encouraged enough to try acetone for exploratory work, and in a pinch (embedding in Paraffin), though I will try to stay with Absolute EtOH for the critical specimens. On these islands, essentially field conditions prevail, and anything I might try will involve making do. This is the spirit in which I had made my query.
(I will take up one kind person's remarkably generous offer to send a bit of Absolute EtOH).
Happy 1998.
Alan Davis
-- "Our loyalties are to the species and the Alan E. Davis planet. We speak for Earth. Our adavis-at-netpci.com obligation to survive is owed not just to Marianas High School ourselves but also to that Cosmos, ancient AAA196, Box 10001 and vast, from which we spring." Saipan, MP 96950 Northern Mariana Islands ---Carl Sagan GMT+10
There is a Unit on our street with the sign "Oxoid" which we always wondered what they did. I took the liberty to ask the employees if they are still in business and could they help you. Their address is Oxoid Inc. 217 Colonnade Road Nepean, Ontario K2E 7K3 Telephone 613 226-1318 or Fax 613-226-3728 or E-mail: Gnyman-at-oxoid.ca A good and prosperous New Year to you all Al Bingham Semoptics Ltd Nepean, Ont
Garry, There is a relatively recent book specifically on microscopy of frozen-hydrated specimens, entitled "Low-Temperature Microscopy and Analysis", by Patrick Echlin, 1992, Plenum Press, New York. It gives a good introduction to the subject. Other than that, my experience was that I had to be very fast on my feet with the one or two samples that could be done in one day, and the long waiting periods while the system pumped down (or not) to ultrahigh vacuum. Good luck with your study.
Does anybody know where I can buy the jewel bearing used in TEM sample rods? I want to make a special sample rod and just require the jewel for the rod.
Many thanks,
Keith.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
If you are after the synthetic sapphire balls used, for example, in JEOL side entry holders we get ours (2mm dia) from: Dejay Distribution Ltd, 9, The Business Centre, Molly Millars Lane, Wokingham, Berkshire RG11 2GS. UK.
However, I am sure that you could find a local supplier for these.
We also use other diameter balls from the same supplier in our specimen holder designs.
Ron ========================================================================== = Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ========================================================================== ==
} } Greetings, } } Does anybody know where I can buy the jewel bearing used in TEM sample rods? } I want to make a special sample rod and just require the jewel for the rod. } } Many thanks, } } Keith. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
There is a company in Manchester UK which manufactures the jewel end bearings. They can aslo make Philips type jewels which are not spherical like the JEOL type. Contact me if you want to know more.
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
I have an InSb [001] sample, and need to know precisely which direction is (100) and which is (010). Does anyone know how to determine this, perhaps using CBED?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Leah L. Dobbs of Intel asked the following:
} Does anyone have experience using the Polaroid Sprint Scan 45 for TEM } negatives? I would appreciate any opinions on this scanner.
About the question of special glass to avoid Newton rings,
We have simply removed the glass on the support for the negatives on our Agfa Duoscan. Our workshop has designed a metal plate drilled with rectangular apertures and frames with the same opening to clamp the negative inbetween. The size of the aperture is large enough to hide only 3mm all around the edge of the negative. We used an aluminum alloy and treated to get it black (to avoid spurious reflections. We do not observe any bending and the negative flatness is sufficient even for 3 1/4" x 4" TEM negatives. Of course don't longer have to worry about fingerprints on the glass window. More, we mount 4 negatives at a time, exactely parralel to each others, it saves prescan time. We are very pleased by this system.
If it helps=8A
Best regards and Happy New Year
Philippe Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
PPG Industries, Inc. Glass Technology Center has a job opening for a Surface
Scientist/Analyst.
PPG Glass Technology Center is located 10 miles north of Pittsburgh, Pennsylvania. The Surface Analysis Laboratory at this site provides surface
chemical and microanalysis to all four divisions of PPG: Glass, Fiberglass, Chemicals, and Coatings and Resins. Currently two major instruments (VG ESCALAB MkII and VG SIMSLAB) operate in five modes for surface analysis: XPS, AES, SNMS, SIMS and FAB-SIMS.
Education: Advanced degree (or equivalent in experience) in chemistry or other related physical science.
Job Requirements: Minimum 3 years of industrial experience collecting and interpreting surface analysis data, preferably XPS and/or SIMS. Data and interpretation of data to be presented in written reports. Good written and
verbal communication skills are a must. Job requires ability to handle multiple tasks simultaneously. Demonstrated practical problem-solving ability is essential.
Detailed information about PPG Industries, Inc. may be found on our Web page
at: http://www.ppg.com.
Send resume (no email, please) and salary requirements to:
Glass Technology Center PPG Industries, Inc. Box 11472 Pittsburgh, PA 15238
We had a machinist at Univ. Florida (Go Gators!) that used to make them from glass (or quartz?). He would take a rod, heat it, and pull to get a thin strand. After breaking the strand, he applied heat and the strand would ball up at the end. When he got the size he wanted, he stopped. The little tail would go into the hole in the end of the specimen rod.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Does anybody know where I can buy the jewel bearing used in TEM sample rods? I want to make a special sample rod and just require the jewel for the rod.
Many thanks,
Keith.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Gary, We have product literature applications notes and a video tape which we can make available to you. Please come back with your mailing address so I can send them off to you. Also there will be a cryo SEM hands on workshop and seminar at Scanning this year. Scanning will be in Baltimore May 9th through 12th. The cryo workshop will be held on Saturday the 9th. Our Cryo product manager will be at the seminar and people involved will have acess to equipment at the USDA in Beltsville Maryland. There will be a limited number of people for the hands on portion (probably around 6 to 8) so if you're interested please get back to us soon.
Feel free to call or me at 978-456-9736. Best regards, Glenn Kinnear
Group, A friend of mine, not on this listserver, would like to contact a company in Germany(?) named ICT. Apparently they are involved in designing, etc. electron optics. Any help, direct to me, would be much appreciated. Don Grimes, Microscopy Today
Crystallographically, [100] and [010] directions are equivalent. You may = arbitrarily choose the coordinate s
} -----Original Message----- } From: L.D.Marks [mailto:ldm2-at-apollo.numis.nwu.edu] } Sent: Monday, January 05, 1998 2:56 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: InSb [001] orientation } =20 } =20 } = ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of = America=20 } To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com } = -----------------------------------------------------------------------. } =20 } I have an InSb [001] sample, and need to know precisely } which direction is (100) and which is (010). Does anyone } know how to determine this, perhaps using CBED? } =20 } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Evanston, IL 60208-3108 } tel: (847) 491-3996 } fax: (847) 491-7820 } email: l-marks-at-nwu.edu } http: //www.numis.nwu.edu } ++++++++++++++++++++++++++++++++++++++++++++++++ }
Crystallographically, [100] and [010] directions are equivalent. You may = arbitrarily choose an appropriate coordinate system to name one = direction [100] and the other [010], etc.
Hartmut
Dr. Hartmut S. Leipner Fachbereich Physik Friedemann-Bach-Platz 6 Martin-Luther-Universitaet D-06108 Halle Germany
Tel. +49-345-55 25 453 Fax +49-345-55 27 563 Web http://www.physik.uni-halle.de/Fachgruppen/Kristall/index.html
} -----Original Message----- } From: L.D.Marks [mailto:ldm2-at-apollo.numis.nwu.edu] } Sent: Monday, January 05, 1998 2:56 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: InSb [001] orientation } =20 } =20 } = ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of = America=20 } To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com } = -----------------------------------------------------------------------. } =20 } I have an InSb [001] sample, and need to know precisely } which direction is (100) and which is (010). Does anyone } know how to determine this, perhaps using CBED? } =20 } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Evanston, IL 60208-3108 } tel: (847) 491-3996 } fax: (847) 491-7820 } email: l-marks-at-nwu.edu } http: //www.numis.nwu.edu } ++++++++++++++++++++++++++++++++++++++++++++++++ }
Dear Garry: A Happy New Year from a dedicated cryo-microscopist to one who intends entering the field. I have been doing cryo-SEM and cryo-x-ray microanalysis for about 25 years. It is a lot of fun. You might care to dip into my book "Low Temperature Microscopy and Analysis"Plenum Press 1992 which will give you some idea of what it is all about.
Patrick Echlin Director, Multi-Imaging Centre School of Biological Sciences University of Cambridge UK
On Fri, 2 Jan 1998, Garry Burgess wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am interested in hearing from someone who has done cryo-electron } microscopy on hydrated samples using a cold stage. I have never done } this myself, and am unable to find much mention of it in the books, but } was wondering what kinds of skills I would have to develop in order to } do this sort of microscopy. } } To all the people who answered my question about CPD, thankyou very } much, it was helpful. } } Garry }
Hello All, I am interested in getting digital images from my rather aged JEOL 120CX TEM/STEM. JEOL advertise a system to do this, which they call SemAfore. I would be very interested in the opinions of users of this system - and those of people who decided to do it some other way, particularly in regard to cost and ease of installation and use. I'll summarise the replies to the list and anyone else who is interested.
Many thanks in advance,
Richard Beanland, GMMT Caswell, Towcester, Northants NN12 8EQ UK e-mail richard.beanland-at-gecm.com
This message is being posted to the: 1. Microscopy 2. Histonet 3. Safety and 4. Sorehand Listservers. It contains two sections. The first is essentially a workstation assessment form for microscope users, the second is an earlier list of instructions for using microscopes safely. These items are based on selections from the many replies to my initial query. Thanks to all those who replied on the Listservers listed above. If you think there is a mistake/problem, please let me know!!
I am also attaching the two sections as Microsoft Word 6 files.
Best wishes for 1998
Keith Ryan Plymouth Marine Laboratory, UK _____________________________________________
Workstation assessment for Safe Use of Microscopes
Introduction. Looking through a microscope for extended periods is not what we were designed for. It requires holding our bodies in an unnaturally rigid position. It is important to adopt a correct, ergonomic working posture. This means fitting the workstation to the worker, not vice versa. It is also important to take regular breaks.
Ideally, the microscope should be on a bench which is adjustable for height: first, the seating position is adjusted (steps 2-8 below) followed by the bench height and subsequent steps (11-22 below). The following is based on a fixed work bench.
Before entering *No' to the following questions, attempt to rectify the problem.
(the following section is a table in the attached file, it looks much better!)
Yes No 1. Have you been show how to use the microscope, including how to align the optical path to optimise performance? If no, seek this training before continuing. Other-wise, poor results and eye strain may ensue.
2. Are you sitting back in the chair, rather than perching on it? If no, sit back into the chair.
3. Is the height of the chair adjusted so that your feet are resting comfortably, flat on the floor? If no, adjust the chair height appropriately.
4. Is there an even pressure along the backs of the thighs? If no, check if the seat platform can be tilted appropriately for comfort. If not, consider readjusting the height slightly.
5. Does the chair support your back in an upright position? Ideally, it should support to beyond the level of the shoulder blades. If no, adjust chair back appropriately.
6. Does the chair back give support in the lumbar region? If no, check the chair's back adjustment again, possibly adjusting the tilt control. Or, obtain a separate lumbar cushion.
7. Are you now sitting with your back upright? Ask a colleague to comment. If no, repeat steps 2-6 above.
8. Are the microscope eye-pieces in line with, or extending over, the front edge of the bench? If no, move the microscope towards you appropriately (caution - you may need skilled assistance)
9. Is the vertical position of the eye-pieces a little high for comfort, so that your head is upright? Initially, this will feel unnatural. If no, raise the microscope vertically to a suitable height at which you are are forced to sit upright. This can be done with layers of plywood etc. If you are using the microscope long-term, get the workshop to make a suitable stand.
10. Can you see at all into the eye-pieces of the microscope? If no, raise the chair height appropriately and obtain a suitable footrest.
11. Are you gazing slightly downwards into the eye-pieces, as opposed to tilting your head and *looking straight-ahead' into them? If no, you are not sitting upright enough. The back should be *vertical' and the neck and head upright. Holding the head tilted for long periods will induce neck and shoulder-ache. Repeat steps 2-10.
12. Is the leg-well clear of clutter so that your legs and feet are not impeded when sitting at the bench? If no, clear the clutter.
13. Are your thighs clear of the under-surface of the bench? If no, the bench is unsuitable for microscopy work. Have it modified or seek another site for the microscope.
14. When operating the focus and stage controls, are your forearms resting on something, either the bench or microscope arm rests? If no, obtain arm rests, perhaps sloping. Holding the arms off the bench for long periods will induce static loading problems. The most comfortable position for the hands is as for when shaking hands.
15. Are the eyepieces set correctly for your inter-pupillary distance? If no, set this distance properly - the oculars should move towards or away from each other. This reduces eye-strain.
16. Are the eyepieces parfocal? If no, adjust them individually so that the image is sharp in each. This reduces eye-strain.
17. Before you begin microscope work for the first time, are you free from pre-existing visual problems? If no, you should see an optician in case you have astigmatism, fusion insufficiency (poor eye co-ordination) or simple long/short sight. Microscopy work may make these problems more obvious.
18. Are your surroundings free from glare and reflections? If no, try to remove light sources from the visual field by re-positioning the workstation, removing highly reflective surfaces, using blinds, curtains or other screens.
19. Is the image in microscope free from glare and reflection? If no, adjust the internal lighting so that there is not an uncomfortably high level of light or contrast. This may be done by regulating the transformer or by using appropriate filters.
20. Are you satisfied with other environmental factors, such as temperature, humidity, draughts, ventilation, ambient lighting? If no, try to sort problems locally yourself or discuss them with line management. Beyond that, see your union safety representative or local safety advisor. Humidity (and dry eyes) are aided by watering plants, where appropriate.
21. Are you/will you be taking regular breaks from the microscope, e.g. two or three minutes every half-hour? If no, this needs urgent consideration. Discuss it with your manager, union safety representative or local safety advisor. Computer users are recommended to take five minutes every hour, microscopy work is probably more physically demanding.
22. When taking a break from the microscope, are/will you be doing stretching exercises? If no, refer to the article *Applying ergonomics to improve microscopy work* in Microscopy and Analysis, Issue 36 (July 1993), 15-17 (available from your local safety advisor). You should do these exercises to relieve the static loading stress on the body. This applies equally to computer users.
_________________________________________________
Instructions for Safe Use of Microscopes
Introduction. Looking through a microscope for extended periods is not what we were designed for. It requires holding our bodies in an unnaturally rigid position. This can cause cramped muscles and strained tendons and ligaments in the head, neck, back, shoulders, arms and wrists. Also, repetitive movements associated with microscope work can cause strain injuries. There are specific health and safety regulations for computer use. Yet, you are tied much more to a microscope than to a computer, because of the eye-piece requirements.
Main problem - microscopists know how to align their microscope but few align themselves! Many users are *slumpers' and need training to avoid problems later. _____________________________________________
Main requirements: The bench height is important and ideally it should be adjustable so that eye-piece height is adjustable - but this is often not practicable due to cost. What follows will be based on a fixed bench height. Microscopy requires a good seating position in an adjustable, ergonomically-designed chair. The back should be high enough to support the shoulder blades and be adjustable for height and angle, with the most prominent part being the lumbar support. The seat should be low enough to have the back *perfectly straight' to see through the oculars. Then move the chair towards the bench so that the chair fully supports the back - this feels unnatural at first. Most people have the seat too high, which results in *hunching'. Sitting for long periods places strain on the lower back.
An alternative chair is the Swedish or Scandinavian chair. First, get the posture right in the chair, then adjust the table/microscope height until the oculars meet the eyes. _____________________________________________
1. Have you been shown how to use the microscope, including the alignment of the optical path so as to optimise performance? If not, stop at this point and seek this training.
2. Adjust chair height so that your feet sit comfortably on the floor. a) get an even pressure along the back of the thighs. b) it may be advantageous to tilt the seat slightly down at the front (to eliminate *pinch' behind the knees). Sit in the chair. Do not tuck the feet underneath the chair or on its base, this tilts the hips away from the backrest. The position of the feet should be varied from time to time, to spread the load on the back and leg muscles. NB - adjust the chair for comfort without regard to the height of the microscope.
Adjust the microscope so that you can see into the oculars without leaning significantly forward. This requires two adjustments: a) set the horizontal position of the microscope so that it is close to the front edge of the bench with the eye-pieces no further away from you than the front edge of the bench b) set the vertical position of the microscope so that it is a little high for comfort. This normally means elevating the microscope on some type of stand on the bench.
The purpose of this is to force yourself to straighten your back as you draw up to the microscope, so that your head is in an upright position. Look down the eyepieces by letting your eyes view at a downward angle. Do not bend your neck so that you are, in effect, looking *straight ahead* into the microscope. Sometimes it is necessary to adjust chair height so that you can just see into the eye-pieces because the bench has a fixed height. Then, you may need a footrest.
The leg-well should be clear. Bench thickness should be minimal to give clearance for thighs - refer to Microscopy and Analysis article.
Obtain forearm-rests so that you do not constantly keep lifting your arms off the bench to adjust the microscope, these may be best if they are sloping. Consider getting a permanent support made for the microscope. It won't cost much and will make an enormous difference to working comfort and productivity. The most comfortable position for the hands is in the mid-range position (as when shaking hands).
Practice continuously focusing - essential to minimise eye strain Proper setting of the inter-pupillary distance and eye-piece parfocality also minimise eye strain.
Take breaks away from the microscope, perhaps ideally every half-hour for a couple of minutes. Get up, walk around, stretch arms, neck, back and legs. _____________________________________________
Appendix - selected information from: Applying ergonomics to improve microscopy work, H. Haines & L. McAtamney, Microscopy and Analysis, July 1993, pages 15-17.
Visual demands Eyestrain is due variously to microscope design parameters, length of work period, pre-existing eye problems and inappropriate lighting conditions.
1. Design of microscopes: beyond the scope of these instructions. See references cited in article.
2. Length of work period: work/rest schedules should be appropriate. It is important to take breaks away from the microscope to counteract the build-up of postural and visual fatigue. A well-designed job will incorporate both microscope and non- microscope tasks. Frequent short breaks are preferable to occasional long breaks. This enables relief from the static, maintained working posture and a chance to look around and vary the accommodation of the eyes. Computer users are recommended to take 5 minutes every hour away from the computer; it may be preferable to take half-hourly breaks away from the microscope. Bending and stretching exercises are recommended (see Microscopy and Analysis article).
3. Pre-existing eye problems: visual strain is pronounced for operators with uncorrected astigmatism and fusion insufficiency (poor eye co-ordination). It is important to treat these problems, see an optician.
4. Inappropriate lighting conditions: eye fatigue is minimised in a well-designed visual environment. Where possible, avoid high light levels and high contrast in the microscope in comparison to the room surroundings. Minimise glare and reflections in the work area. Glare can be reduced by removing light sources from the visual field. This can be done by re-positioning workstations, using blinds or curtains, removing highly reflective surfaces or by using shielding screens. The Health & Safety (Display Screen Equipment) Regulations 1992 give further information regarding work with VDU screens.
Environmental factors Sedentary workers are particularly susceptible to the effects of their work environment. Draughts, temperature extremes, poor air quality, inadequate lighting and noise affect comfort and performance. It is important to consider the needs and preferences of the individual.
Temperature, humidity and air movement: the recommended range is 19-23 *C. Low humidity dries the eyes and produces discomfort, relative humidity should be maintained between 40 and 60% (watering plants fulfils this need in offices). Draughts should be minimised.
END OF MESSAGE
++++++++++++++++++++++++++++++++++++++++++++++++++ Keith Ryan B.Sc., Ph.D., C.Biol., M.I.Biol. Plymouth Marine Laboratory, Citadel Hill, Plymouth, Devon PL1 2PB, England
Thanks for people who have sent responses -- but to date no solutions. Let me expand a little with a diagram to clarify. Looking down [001] the structure is:
IN SB IN SB --} [110]
Sb In Sb In
IN SB IN SB
| V [1,-1,0]
Where, along z or [001] the order is IN, SB, In, Sb (Note: I may not have picked a proper right-handed set but this does not matter.)
Indium and Antimony have similar structure factors (seperated by 2 in the periodic tables) although the Debye Waller factor for In is about 80% higher than for Sb (Saravana, Mohanlal and Chandrasekaran, J Phys Chem Solids 52, 7, 879, 1991). The problem is to align the above with a diffraction pattern. Almost certainly optical methods will work, but the probability of correctly aligning this with a diffraction pattern is small (particularly since we have to do other things to the sample).
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
I came across this reference today in a lit. search. Perhaps it might be interesting to you. Best wishes, Janine Sherrier
TI- NEW IMMUNOGOLD PROBES FOR STUDYING THE DISTRIBUTION OF THE DIFFERENT LIGNIN TYPES DURING PLANT-CELL WALL BIOGENESIS AU- RUEL, K;FAIX, O;JOSELEAU, JP JN- JOURNAL OF TRACE AND MICROPROBE TECHNIQUES PY- 1994 VO- 12 NO- 4 PG- 247-265
A couple of years ago I ran into the problem of finding a 1 mm diameter ball for use in the tip of a side-entry specimen rod I was designing. I wasn't able to find a sapphire ball to fill the bill, but did find that a local company, Industrial Tectonics, Ball Division, 7222 West Huron River Drive, Dexter, MI 48130, tel: 734-426-4681, makes precision balls of various sizes out of a range of other wear-resistant materials. In the end I was able to obtain 1mm tungsten carbide balls that served the purpose quite well (these are used in ball point pens, and so you might be able to just dissect a pen). (Incidentally, I did not find their sales people too helpful in being willing to supply just a few balls, they wanted to sell a minimum of several hundred dollars worth; therefore, I suggest you try to get in touch with someone in engineering, who might send you a few as a scientific sample). I still have a few of these balls left - if you want, I'll be happy to send you a couple. They also make larger balls out of similar materials, but not sapphire.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Does anyone know of an atlas of mouse histology? With the ultrastructural work we are doing lately with transgenic mice, I must admit that I am not always sure that the interesting structures we see are normal or result from genetic manipulation. Obviously, we observe control tissue, but an atlas would be invaluable as a baseline for comparison.
Thank you in advance,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
Please forward to me directly the current phone or email address for Alle= n R. Sampson. Allen used to service Etec SEMs and other electronics equipment.
TIA, Steve Miller RMC Tucson, AZ Steve.Miller-at-RMC-Scientific.com
I have been looking for a used phototube that will fit a Wild M7A or M3Z stereomicroscope. Any phototubes bearing one of the following Wild/Leica part numbers will do: #10 446 174, #10 445 924 or #10 445 925. Although I would prefer a good quality, used Wild/Leica product, I am willing to consider a Chinese manufactured tube so long as it is compatible with my Wild/Leica instruments.
I am also in the market for an inclined binocular tube that will fit either the M7A or M3Z. Again, these can bear any of the following part numbers: #10 445 619, #10 445 822, #10 429 781 or #10 429 782
Please note, parts that fit the Wild M5A instrument are not interchangable with the rest of the Wild M series stereomicroscopes.
Semafore is less powerful than others. We can do very few application from this solfware.
Member
On Tue, 6 Jan 1998, Richard Beanland +44 1327 356363 wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello All, } I am interested in getting digital images from my rather aged JEOL } 120CX TEM/STEM. JEOL advertise a system to do this, which they call SemAfore. } I would be very interested in the opinions of users of this system - and } those of people who decided to do it some other way, particularly in regard to } cost and ease of installation and use. I'll summarise the replies to the list } and anyone else who is interested. } } Many thanks in advance, } } Richard Beanland, } GMMT Caswell, } Towcester, } Northants NN12 8EQ } UK } e-mail richard.beanland-at-gecm.com } }
I am not sure that what I am going to say to you will be adapted to your situation. If I got it rigth, you want to distinguish the B=[011] direction from a A=[1-10] direction in a bulk InSb material.
There should be a solution to this problem because some people know how to prepare controlled vicinal surfaces in one of these two directions. Myself, I do not know how they do, but once multilayers have been grown on these vicinal or nominal (001) surfaces we are able to determine which direction is which by observing {110} cross-sections of the multilayers by High Resolution Electron Microscopy. This work has been part of Marion Charleux's PhD work (submitted the 1/4/97 at the Joseph Fourier University, Grenoble France) and has been submitted to JAP. It has not been accepted for publication yet.
If you are interested by these methods, answer me.
PS What is the difference between Sb and SB in your scheme ? -- ++++++++++++++++++++++++++++++++++++++++++ Jean-Luc ROUVIERE CEA-GRENOBLE 17 rue des Martyrs 38054 Grenoble Cedex 9 - France Tel. (33) (0)4 76 88 50 86 Fax. (33) (0)4 76 88 50 97 jrouviere-at-cea.fr ++++++++++++++++++++++++++++++++++++++++++
The visual info I am sometimes looking for (good EM pictures of this or that virus) being scattered among a lot of books I have access to, I was wondering whether there is/are recent and well illustrated atlas(es) on virus ultrastructure. I have Virus Morphology but it seems a bit outdated in light of the recent advances in the field, particularly with the data obtained with cryo-EM.
Any recommendations?
Thanks a bunch in advance. Michel
**************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
On behalf of a colleague (please reply to fkoch-at-csir.co.za)
We are trying to obtain electron transparent ( 120 kV) single crystalline (or large grain) Co3O4 films to be used as substrates for subsequent epitaxial growth. Our facilities to thin bulk specimens are limited and we are forced to used a vacuum evaporation/oxidation route or something similar like the MoO3 sublimation route. So far we tried growing epitaxial Co on Ag on mica, a la Grunbaum, and do get single crystals but do experience other problems. The Co film removal for TEM is not simple and oxidation, by heating in air, of the crystals results in the formation of a finely divided oxide which is also not desirable. Any suggestions will be much appreciated.
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Please forward to me directly the current phone or email address for Allen R. Sampson. Allen used to service Etec SEMs and other electronics equipment.
TIA, Steve Miller RMC Tucson, AZ Steve.Miller-at-RMC-Scientific.com
by rks3 with SMTP (PP); Wed, 7 Jan 1998 14:02:47 +0100 Received: from rcs1.urz.tu-dresden.de by rmail with SMTP (PP); Wed, 7 Jan 1998 14:00:23 +0100 Received: from [141.30.35.16] by rcs1.urz.tu-dresden.de (8.6.12/SDL-5.8) with SMTP id OAA27901; Wed, 7 Jan 1998 14:02:42 +0100
Dear Dough,
there is an excellent atlas about mouse development (mostly histology, some scanning EM, few transmission EM):
Matthew H. Kaufman "The atlas of mouse development", Academic Press, London/ San Diego (1st edition in 1992, revised ed. in 1995).
Best wishes
Heinz
*********************************************************************** Dr. Heinz Fehrenbach Institute of Pathology University Clinics "Carl Gustav Carus" Technical University of Dresden
SALZBURG, Austria, 7th of Jan, 1998, local time: 3.20 p.m.
First of all: Happy, HEALTHY, SUCCESSFUL NEW YEAR to YOU and YOURS!
Secondly: have seen your posting and Query: if the request hasn't been answered in the meanwhile, here you go:
- The Company "ASSISTENT" doesn't exist: it is a "Brand". - The fabricating company is:=20 Karl HECHT GesmbH & CoKG Stettener Strasse 22-24 D-97 647 SONDHEIM/Rhoen
for a contact outside USA, if you can't get what you want from:
CAROLINA BIOLOGICAL 2700, York Road BURLINGTON, NC 27 215-3398 USA
(only) FaxNo (available): USA: (919) -584-33-99
Hope this helps, Best regards and a nice day
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: Eric or Pat Metzler[SMTP:spruance-at-infinet.com] Gesendet: Samstag, 03. J=E4nner 1998 02:30 An: Microscopy-at-sparc5.microscopy.com Betreff: Q: "Assistent" company in Germany /MSA: Search for Adresses, = LM: Sectioning:cover glass
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I have some cover slides made by a company named ASSISTENT. According = to the box, the company is (was) located in Germany. The cover slides are = very small (5 mm to 8 mm diameter) and are ideal for one of my uses. I'd = like to locate this company, a replacement, or anyone else making such a = product. Can you please help me?
Reply directly to me if you do not want to reply to everyone. spruance-at-infinet.com
They have small balls made of Aluminum Oxide, Synthetic Ruby and Synthetic Sapphire (as well SS, Brass, Tungsten Carbide, Plastics, Teflon, etc.,, etc.). Qunatities of (1) on up in sizes: of 1/64", 1/32", 1/16", 3/32", 1/8" [1/4", 1/2" in AlO], sphericity of 0.000025" on Synthetics and 0.00001" on ALO. At ~$4.00 each for the jewels and $3.00 for most of the ALO's (in single quantities).
Small Parts has changed policy and there no longer is a min order $.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
email ars-at-mcs.net
ph# 708-513-7093
fax 708-513-7092
http://www.mcs.net/~ars/home.html
Maya Moody Cell Imaging Facility Cell and Molecular Biology Northwestern University Medical School Ward-7-143 303 E. Chicago Ave. Chicago, IL 60611 T: 312-503-4445 F: 312-503-7912 E: m-moody-at-nwu.edu
} } -----------------------------------------------------------------------. } } } } Please forward to me directly the current phone or email address for Allen } } R. Sampson. Allen used to service Etec SEMs and other electronics } } equipment. } } } } TIA, } } Steve Miller } } RMC } } Tucson, AZ } } Steve.Miller-at-RMC-Scientific.com } } } } }
I must justify the need for FE-SEM or TEM capabilities to a customer wanting to characterize particles smaller than 0.1 microns. (We presently have a conventional SEM). I would like to include a photomicrograph or two in my report of ultrafine particles imaged by FE-SEM demonstrating the resolution capabilities of FE-SEM. If you are willing to share one of your images, please correspond to the email address below (not to the Listserver please!). A TIFF file accompanied by a brief description would be great. Thank you very much.
Bob Willis ManTech Environmental Technology, Inc e-mail:willis.robert-at-epamail.epa.gov
Happy New Year, Doug - My brother-in-law was looking over my shoulder this morning when your request for a mouse histiolgy atlas arrived. He teaches histology & has done a bit of rodent research & he suggests a book from the 60s on mouse embryology by Roberts Rugh. Happy hunting...
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Greetings this is my first post to this list, I am looking for coverslips or slides that will enable me to record the exact location of an object (calcite crystal) when viewed with a light microscope. I then would like to image the same crystal with an SEM, to view the morphology of the crystal. I have heard of slides or coverslips with grids that are labeled for just this purpose. I have not been able to find slides or coverslips for sale in any catalogs or on the web. I have also searched the archives with no luck. Perhaps someone has had similar experiences or knows just just what I am looking for. If so please let me know. TIA thom
________________________________________________________________________ Thomas Geer Marine Biological Laboratory Woods Hole, MA 02543 Tel:508-289-7629 fax:508-540-6902 E-mail:tgeer-at-mbl.edu web http://www.mbl.edu/oldenbourg
Dear Richard, I don't believe the JEOL SemAfore system will digitize a TEM image, I think it is just for SEMs. Any time you want to digitize a TEM image, which is a real image as opposed to a raster, you must use some sort of camera. It is the interfacing of the camera to the TEM that is the expensive part, and the camera, if you want better than video resolution, which is quite poor. The STEM can go in just like an SEM. The output of the camera can, perhaps, go into the SemAfore, or any other system, such as Quartz PCI. I think the expert on putting cameras on TEMs is Jim Mancusso, of AMT. The phone number is +1 508 774 5550. (USA)
You wrote: } Hello All, } I am interested in getting digital images from my rather aged JEOL } 120CX TEM/STEM. JEOL advertise a system to do this, which they call SemAfore. } I would be very interested in the opinions of users of this system - and } those of people who decided to do it some other way, particularly in regard to } cost and ease of installation and use. I'll summarise the replies to the list } and anyone else who is interested. } } Many thanks in advance, } } Richard Beanland, } GMMT Caswell, } Towcester, } Northants NN12 8EQ } UK } e-mail richard.beanland-at-gecm.com } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Concerning a book by Rugh Roberts, a search using the internet turned = up this book.
Book Details for ISBN no. 0198542771
Mouse, The: Its Reproduction and Development by Rugh, Roberts, Price: =A367.95 Hardback, 434 pages, Publisher: Oxf. University Press, Date Published: Apr 1990
Etched grid coverslips or slides are not an option if both LM and SEM are to be used.
One must mark the sample directly. Inked circle markers are available from Jaynes-Elkro of Columbus, Ohio. Phone 614 276 6196 (in 1992).
OR.. I supply the Microscope Marker / Drill which mounts in place of an objective lens in a light microscope and turns the function of the microscope racking system into a little drill press. 60 um holes can be drilled or 500 to 1500 um circles can be scribed.
Bart Cannon Cannon Microprobe 1041 NE 100th St. Seattle, WA 98125 206 522 9233 cannonmp-at-accessone.com
I like to announce that FEI Company's software development group for Electron Microscopes in Eindhoven, The Netherlands, has several jobopportunities. Please look at our Website http://www.peo.philips.com/jobmain.htm for more details.
Lawrence, the previous replies you had were right, if your InSb has the usual sphalerite (zinc blende) cubic structure (and if it doesn't - wow!). The symmetry of the crystal is Fbar43m; although there is no four fold axis, there is a bar4 axis (4 fold plus inversion). So [100] is equivalent to [010]. Any textbook on crystallography will show this - my favourite is 'Crystallography and Crystal Defects', by Kelly and Groves.
Regards,
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK e-mail richard.beanland-at-gecm.com
} Thanks for people who have sent responses -- but to } date no solutions. Let me expand a little with a } diagram to clarify. Looking down [001] the structure } is: } } } IN SB IN SB --} [110] } } Sb In Sb In } } IN SB IN SB } } | } V } [1,-1,0] } } Where, along z or [001] the order is IN, SB, In, Sb } (Note: I may not have picked a proper right-handed set } but this does not matter.) } } Indium and Antimony have similar structure factors (seperated } by 2 in the periodic tables) although the Debye Waller factor } for In is about 80% higher than for Sb (Saravana, Mohanlal and } Chandrasekaran, J Phys Chem Solids 52, 7, 879, 1991). The } problem is to align the above with a diffraction pattern. } Almost certainly optical methods will work, but the probability } of correctly aligning this with a diffraction pattern is } small (particularly since we have to do other things to the } sample). } } } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Evanston, IL 60208-3108 } tel: (847) 491-3996 } fax: (847) 491-7820 } email: l-marks-at-nwu.edu } http: //www.numis.nwu.edu } ++++++++++++++++++++++++++++++++++++++++++++++++
I have the following atlas, although I do no know whether it still is in print:
William D. Gude, Gerald E. Cosgrove, Gerald P. Hirsch. 1982. Histological Atlas of the Laboratory Mouse. Plenum Press, New York.
ISBN 0-306-40686-1
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
I hope all you "listees" have had happy holidays...
Now to my question:
We've been using fast drying conductive silver paint to contact the top of metallized plastic film samples to the stub for imaging with our SEM (W filament). Usually this works all right, but recently we've been having problems.
The conductivity is fine at first, but after a while (about a day or so) it degrades and we get charging. We know its not the sample or the metallized coating that's degrading or aging because when we put a fresh dab of paint on, things are OK again. This aging seems to take place both in the vacuum chamber and outside.
Is this normal? Is there some paint out there that doesn't age? Or are there better ways of contacting upper conductive layer on our samples to the stub?
I think the answer is that you are both correct. It doesn't matter if I call my surface normal [001] or [100] as the two are equivalent. But once I choose to call my surface [001], it DOES matter which direction I call [100] and [010]. This is because I want (001) + (010) + (100) = (111)A (i.e. a group III terminated (111) plane or a group V plane if you defined your basis the other way). The point is is that you need to keep track of the [110] and [1-10] directions on a III-V(001) cut wafer (or the [011] and [01-1] on a (100) wafer ...) as the two direction are not equivalent.
I hope this helps, -- Ray D. Twesten Center For Microanalysis of Materials (217) 244-6177 University of Illinois (217) 244-2278 (fax) 104 S. Goodwin Ave. (217) 359-4035 (home) Urbana, IL 61801 http://www.mrl.uiuc.edu/~cmm/index.html
I would like to 'talk' to anyone who has experience with using NIH Image to quantify the amount of antibody staining in LM sections (by optical densitometry). I am particularly interested in methods of calibration. I have a number of constraints, so I think this will be a tough one.
Please e-mail me at tina-at-pbrc.hawaii.edu
Thanks! Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Many thanks to those who responded to my query about=20 Histology books relating to the mouse. For those also=20 interested, here are all three suggested references:
Matthew H. Kaufman "The atlas of mouse development",=20 Academic Press, London/ San Diego (1st edition in 1992,=20 revised ed. in 1995).
Robert Rugh "The Mouse: Its Reproduction and Development"=20 Oxford University Press, Date Published: Apr 1990. =20 Hardback, 434 pages, =A367.95 ISBN no. 0198542771
William D. Gude, Gerald E. Cosgrove, Gerald P. Hirsch. =20 1982. "Histological Atlas of the Laboratory Mouse." Plenum=20 Press, New York. ISBN 0-306-40686-1 ----------------------=20 Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
Does anyone out there have a protocol for IHC on frozen rat brain sections ? We're planning on looking at cytokine expression. Robin E. Neft, Ph.D. Lovelace Respiratory Research Institute P.O. Box 5890 Albuquerque, NM 87185 RNEFT-at-lrri.org FAX: 505-845-1198 505-845-1142
Since silver oxide is sufficiently conductive, oxidation should not be the problem. Micro-cracking of the adhesive or debonding of the interface would cause such a problem. Not knowing materials, can't be much direct help. I am using carbon paint rather than silver. The conductive particles are MUCH smaller, and carbon does a good grounding job for EM. Also, it is usually cheaper.
Woody White McDermott Technology, Inc. ______________________________ Reply Separator _________________________________
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Hello there microscopy world!
I hope all you "listees" have had happy holidays...
Now to my question:
We've been using fast drying conductive silver paint to contact the top of metallized plastic film samples to the stub for imaging with our SEM (W filament). Usually this works all right, but recently we've been having problems.
The conductivity is fine at first, but after a while (about a day or so) it degrades and we get charging. We know its not the sample or the metallized coating that's degrading or aging because when we put a fresh dab of paint on, things are OK again. This aging seems to take place both in the vacuum chamber and outside.
Is this normal? Is there some paint out there that doesn't age? Or are there better ways of contacting upper conductive layer on our samples to the stub?
id AA27244 for Microscopy-at-Sparc5.Microscopy.Com; Thu, 8 Jan 98 15:27:33 MST Received: from email56.sps.mot.com by mogate.sps.mot.com (4.1/SMI-4.1/Email-2.0) id AA04417 for Microscopy-at-Sparc5.Microscopy.Com; Thu, 8 Jan 98 15:26:46 MST Message-Id: {n1327849404.28889-at-mesaqm.sps.mot.com}
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Reply to: RE} } how to tell orientations in InSb [001]
Well, It seems that everyboday is correct here. (001) III-V materials usually have a cutoff angle several degrees toward one of the {110 } directions. There is a differece between two crystalllogrrphically equivalent planes (110) and (1 -1 0) because one plane is parallel to the wafer normal but the other one is several degree off the wafer normal. (Important!! wafer normal is several degree off the (001) plane)
If you have a whole wafer, the major flat should parallel to the (110) planes but the minor flat will be several degree off the crystallography (1-10) plane.
If you only have a small piece of a wafer and it is cleaved, you can still tell. Becasue III-V material usually have (110) type cleavage planes, the cleavage plane should follow the crystallography. Therefore, one of the cleavage plane will be perpendicular to the wafer normal and the other one will not. This can be check under SEM or may be a high mag optical microscope.
Tan-Chen Lee Motorola, Inc. -------------------------------------------------------------------- I think the answer is that you are both correct. It doesn't matter if I call my surface normal [001] or [100] as the two are equivalent. But once I choose to call my surface [001], it DOES matter which direction I call [100] and [010]. This is because I want (001) + (010) + (100) = (111)A (i.e. a group III terminated (111) plane or a group V plane if you defined your basis the other way). The point is is that you need to keep track of the [110] and [1-10] directions on a III-V(001) cut wafer (or the [011] and [01-1] on a (100) wafer ...) as the two direction are not equivalent.
I hope this helps, -- Ray D. Twesten Center For Microanalysis of Materials (217) 244-6177 University of Illinois (217) 244-2278 (fax) 104 S. Goodwin Ave. (217) 359-4035 (home) Urbana, IL 61801 http://www.mrl.uiuc.edu/~cmm/index.html
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I am an undergraduate student doing bacteriophage research. I was wondering if anyone knew of a good, inexpensive EM server in the Philadelphia area, or of any servers where I could send my sample to be done. If one comes to mind, please let me know. Thank you, Emily
I am forwarding the following query for a colleague. Please reply to her at mcfallng-at-hawaii.edu
Mahalo! Tina ****************************************************************************
Dear Microscopists-
In purchasing a confocal microscope, we'd like to get some input from current users of Zeiss and Leica confocal scopes to get an idea of user satisfaction.
Many thanks for any and all help in making our selection.
Margaret McFall-Ngai Pacific Biomedical Research Center University of Hawaii
Dear Cindy, I have the same problem sometimes with the carbon dag I use. I believe it is a small crack appearing in the paint, too small to see, as the paint becomes brittle upon further drying. Using copper tape (in most EM catalogues) may solve this, or, as you say, another small dab of paint. You wrote: } Hello there microscopy world! } } I hope all you "listees" have had happy holidays... } } Now to my question: } } We've been using fast drying conductive silver paint to contact the } top of metallized plastic film samples to the stub for imaging with our } SEM (W filament). Usually this works all right, but recently we've been } having problems. } } The conductivity is fine at first, but after a while (about a day or so) } it } degrades and we get charging. We know its not the sample or the } metallized coating that's degrading or aging because when we put } a fresh dab of paint on, things are OK again. This aging seems to } take place both in the vacuum chamber and outside. } } Is this normal? Is there some paint out there that doesn't age? Or } are there better ways of contacting upper conductive layer on our } samples to the stub? } } TIA! } } Cindy Bennett Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
We have got a recurrent problem in the vacuum system of our CM30. When P1 (pirani head, reading pressure in the vacuum buffer) reaches 38, the P3 (penning gauge, reading pressure in the lower chamber) pressure increases and the vacuum system goes into "startup" status until the pump empty again the buffer down to 30. Then the P3 pressure goes back to reasonable values, about 55. Actually it goes back to normal values already while pumping when P1 reads 37. On the other hand, while in the past the P3 could get down to 34 (or even "0", as no reading is available below "34"), it does not any more. We have changed the oil of the ODP and noticed some improvements, but the problem is not completely fixed.
Any hint is welcome. I suspect problems with the reading of P1, giving a pressure lower than the real one, which might prevent the OPD from working in suitable conditions. However, I have no experience in that matter.
thanks
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Lluis Sole i Sabaris, 1-3 E-08028 BARCELONA ESPANYA
If the paint is really fast drying, maybe it is too thin (i.e. too 'watery' or 'runny')?
The only time I have had this type of charding problem (over too many years to mention), it has been because of movement of the specimen by accidentally touching it or by hitting it on the side/roof of the airlock going in or out of the microscope. Is it that your specimens might be heating a little and that expansion is breaking the conductive path? I can't believe that silver itself can actually 'age' to give the efect you describe! Unless it is very thinly applied and can oxideise.
I would suggest a slightly 'thicker' solution of paint, or try two or three applications in sequence. That is, one coat, dried followed by a second and maybe a third after the second is dry? I am curious.
Finally, if you have any old silver, or gold, or diamonds, that have aged and are no longer any good, you can send them to my address at ...... !
Cynthia- We have switched from silver paint to collodial graphite paint for making contact to SEM stubs. It works well and is much easier to apply. Mike mcalabr-at-rsc.rockwell.com
test message to check subscription -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Hello to all!!! I'm in my last year of the electron microscopy program at S.J. Delta College. I would like some information or ideas on who to contact for summer internships that deal with electron microscopy (FIB,SEM,TEM,OLM, etc......) as well as any sample prep. work primarily in the Materials field (Semiconductors). Thank You.
This is a common protocol to be found in IHC texts and materials and methods sections of numerous papers. An easy start would be the protocols offered in the rear of the Immunochemicals section of your Sigma catalog.
Steve Samuelsson ______________________________ Reply Separator _________________________________
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Does anyone out there have a protocol for IHC on frozen rat brain sections ? We're planning on looking at cytokine expression. Robin E. Neft, Ph.D. Lovelace Respiratory Research Institute P.O. Box 5890 Albuquerque, NM 87185 RNEFT-at-lrri.org FAX: 505-845-1198 505-845-1142
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Fellow microscopists,
I am aware of a 3mm titanium grid for TEM specimens which allows you to put a specimen in a little slot inside it, and then to put a little screw- driver in a parallel slot on either side of the specimen and twist a little to hold the specimen in place, but I am having difficulty locating a manufacturer. Any help finding a supplier would be greatly appreciated. Thanks very much for your time and help.
Sincerely,
Mick Thomas Cornell Materials Science Center Cornell University Ithaca, NY 14853 607-255-0650 mgt3-at-msc.cornell.edu
At 11:15 AM 1/9/98 -0400, David Wark wrote: } I'm planning to purchase a one-inch mount containing a variety of metals - } the more the merrier. I'd be interested in learning the names of vendors } supplying such mounts...
We (Energy Beam Sciences) have, for many years, sold microprobe standards made by Micro Analysis Consultants in the U.K. The list of available standards can be accessed from our web site (http://www.ebsciences.com). Multi-element standards can be mounted in 25mm or 32mm brass blocks.
Best regards, Steven E. Slap Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
We have a Kevex 7000 EDS system (purchased in 1978) which we'd like to donate to a school. As far as I know, it still works (no guarantees). We had it connected to an Hitachi S-530 scanning electron microscope so it would fit a similar instrument. Please contact me by email or phone if you're interested. The system is located in Albany, CA (Berkeley) so we'd prefer someone who can just come and pick up the system.
Thank you.
*****************************************
Delilah F. Wood United States Department of Agriculture Western Regional Research Center 800 Buchanan Street Albany, CA 94710
Hello Hank, I routinely use formvar coated slot grids(1mm/2mm oval slot) for mounting ultra thin sections. I do not use carbon coating, however. I hold the grid with forceps, immerse the grid in the water in the diamond boat, approach the section at a 45 degree angle. Allow the edge of the section to adhere to the formvar, then gently withdraw the grid from the water at a 45 degree angle. That should do it. I set the grids on a copper plate with 3mm holes . The grids perch on top of the holes till they are dry.
Good luck.
Sally Shrom
On Fri, 9 Jan 1998 hadams-at-NMSU.Edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anyone have any suggestions for mounting ultrathin sections on } to carbon-coated, formar slotted grids? } TIA } } Hank Adams } EML } New Mexico State University }
Dear Sir, Here at Electron Microscopy Sciences we sell MicroProbe Standards-All types and varieties. You may look at our list of standards on our website at emsdiasum.com or in our catalog. Please let us know if you have our catalog and if not we would be more then happy to send you one. Sincerely,
Does anyone have some suggestions for mounting ultrathins on to carbon coated, formar covered slotted grids and staining them? Hank Adams EML New Mexico State University
I am relaying this message for a student who has a problem with specimen prep. Please reply to the following address since he is not on this server. skaza-at-siu.edu Thanks.
Hi all,
I'm looking at submicron ceramic powders (SiC, Si3N4, TiC etc) under the TEM. But I cannot get morphological information from the specimens I've prepared due to particle agglomeration. The specimen preparation technique I've used is:
Making a suspension of the ceramic powders in Iso-Butanol. Sonicating them in a water-bath type sonicator for 90 minutes to 3 hrs. And then dropping the suspension on the grids by means of a pippet. I used about 3-4 drops each time. But the sonication does not seem to break up the agglomeration present. I've also tried microprobe sonicator (sonicating for 30 seconds). That doesn't seem to help either.
My questions are:
1) Does sonicating for a longer time in water-bath type sonicator or a microprobe sonicator help? If so, How long for each?
2) Does milling of the powders in a ball mill type milling container help? Doesn't this end up breaking the particles along with the agglomerates?
3) Does the use of a different suspension medium like methanol or ethanol alleviate the problem, if not solve it?
Any suggestions or references would be appreciated.
We seem to have lost our Stahl motorized stage documentation. Would anyone be able to share the communication codes with us? (Stahl Model 517 MF X Y Micro Position Controller).
Thankyou
Cynthia J. Zeissler Physical Scientist National Institute of Standards and Technology cynthia.zeissler-at-nist.gov 301-975-3910
Announcing the Monthly Meeting of the San Francisco Microscopical Society 16 January 1998 Rockridge Branch, Oakland Public Library 7:00 PM
The January meeting of the SFMS is the annual business meeting where officers will be elected and a revision of the Constitution and By-Laws will be discussed. After the business meeting will be our regular technical meeting. The topic this month is the New PARTICLE ATLAS Electronic Edition. The speaker will be Stephen A. Shaffer.
(Hey! That's me!)
For further particulars, please see the SFMS web page at:
Thanks and I hope to see many of you there. -- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
Dr. Cynthia Bennett wrote: =============================================== We've been using fast drying conductive silver paint to contact the top of metallized plastic film samples to the stub for imaging with our SEM (W filament). Usually this works all right, but recently we've been having problems.
The conductivity is fine at first, but after a while (about a day or so) it degrades and we get charging. We know its not the sample or the metallized coating that's degrading or aging because when we put a fresh dab of paint on, things are OK again. This aging seems to take place both in the vacuum chamber and outside.
Is this normal? Is there some paint out there that doesn't age? Or are there better ways of contacting upper conductive layer on our samples to the stub? ==================================================== In the general case, what you are experiencing is not "normal".
I have personally been involved with the testing and formulation of both silver and then carbon paints (and pastes) for SEM applications for many years. And while silver (or carbon) paints might have the "appearance" like they were all the same, I can state with certainty that they are not all the same. They differ not only only in terms of the particle size and the particle degree of dispersion, and the percent of silver (or carbon) solids, but also in terms of the carrier (check the MSDS sheets) and also the presence of polymer binders and if they are present, then even down to the composition of the polymer. Some paints might indeed have no polymer present.
When there is strange behaviour of the type described, one should keep in mind that one of the two most important properties or characteristics that a formulation should possess has to do with the uniformity of the drying of the suspension, not only into a uniformly thick and homogeneous layer, but it should dry in a way in which there is a minimum of "skin" effect, that is , when the paint forms an undesirable "dry" outer layer ("skin") that acts as a diffusion barrier for the vapor still being evolved from the still "wet" paint suspension underneath. These are also exactly the conditions where a "mud cracking" phenomenon can be observed,something occurring in the skin layer, as it tries to contract over the still "wet" core.
While any paint from any source can be made to not form a skin (and crack) by diluting it down with (its own) diluent, it also loses its adhesive if not also its electrical characteristics. And by the same token, evaporate away quickly enough of the carrier from any of these paint products, you will eventually cause any product to form a skin if drying proceeds too quickly and one will eventually end up with a mud-cracking effect (with the release of vapor into the vacuum of the SEM) if the drying is done quickly enough (such as premature insertion into a vacuum).
So other than the obvious (try some other brand of paint), try diluting down your paint and/or switching from the rapidly drying type to the more conventional drying paint product. See if that does not make your problem go away.
Disclaimer: SPI Supplies formulates and offers for sale silver and carbon paints, pastes, tapes and sheets, all of which are described on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We have the titanium "grids" you describe. They were actually developed=
by Dr. Arpad Barna at the Research Institute of Technical Physics in Hungary. These grids are used in conjunction with his TEM sample preparation methods that he developed for use with the IV3 Ion Mill. We have these sample holders in the following sizes:
Part# Description TL-TiS-06 Titanium Sample Holder, 3mm OD, Center Slot 0.6mm= x 1.8mm TL-TiS-08 Titanium Sample Holder, 3mm OD, Center Slot 0.8mm= x 1.8mm TL-TiS-10 Titanium Sample Holder, 3mm OD, Center Slot 1.0mm= x 1.8mm
Keep in mind that these are not "grids" as such, but rather a 0.3mm thick=
"clamp" for holdng together cross section samples without using epoxy. F= or additional information on the titanium sample holders or the IV3 Ion Milling System, please feel free to contact me.
1. I have done a lot of immunolabelling of TISSUE sections both for EM and light microscopy. However, now in my new job I am faced with a problem of doing immunohistochemistry on cell cultures. Could somebody suggest a good book on cell preparation; such as fixation, suitable substrates for growing cells that allow subsequent immunowork, treatment of slides for cytospins etc? Frankly, I dislike the look of trypsinized and cytospinned cells with very little identifiable morphology and the fact that there is no inherent control in a homogeneous cell population as in tissue sections, where some types of cells are expected to label while others are not.
2. Could somebody give me directions how to subscribe to HistoNet or any other histochemical or immunohistochemical list?
Thank you for your answers,
Sarka Lhotak Hamilton Cancer Center Hamilton, Ontario, Canada lhotaks-at-mcmaster.ca
While this isn't a very cost effective solution for you in this case, it may be interesting to know that RJ Lee Instruments makes an SEM that has = a facility to view the sample under an optical microscope and then transfer=
the sample to their SEM while imaging the same location. I don't know mu= ch about the details, but it may be interesting to talk to them if this is a=
situation you encounter on a regular basis. You can contact them directl= y at:
R.J. Lee Instruments 515 Pleasant Valley Road Trafford, PA 15085 =
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy. Message text written by INTERNET:tgeer-at-mbl.edu } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Greetings this is my first post to this list, I am looking for coverslips or slides that will enable me to record the exact location of an object (calcite crystal) when viewed with a light microscope. I then would like to image the same crystal with an SEM, to view the morphology of the crystal. I have heard of slides or coverslips with grids that are labeled for just this purpose. I have not been able t= o find slides or coverslips for sale in any catalogs or on the web. I have also searched the archives with no luck. Perhaps someone has had similar experiences or knows just just what I am looking for. If so please let me=
know. TIA thom
________________________________________________________________________ Thomas Geer Marine Biological Laboratory Woods Hole, MA 02543 Tel:508-289-7629 fax:508-540-6902 E-mail:tgeer-at-mbl.edu web http://www.mbl.edu/oldenbourg {
Sarka: Go on our site and find the "Links". User the browsers Control F function and search for histonet. Most microscopy pertinent listservers are listed with instructions on how to subscribe. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: Sarka Lhotak {lhotaks-at-fhs.csu.mcmaster.ca} } To: Microscopy-at-sparc5.microscopy.com } Subject: immuno of cell cultures+HistoNet } Date: Sunday, 11 January 1998 10:51 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello Listers, } I would like to ask two questions: } } 1. I have done a lot of immunolabelling of TISSUE sections both for EM and } light microscopy. However, now in my new job I am faced with a problem of } doing immunohistochemistry on cell cultures. Could somebody suggest a good } book on cell preparation; such as fixation, suitable substrates for growing } cells that allow subsequent immunowork, treatment of slides for cytospins } etc? Frankly, I dislike the look of trypsinized and cytospinned cells with } very little identifiable morphology and the fact that there is no inherent } control in a homogeneous cell population as in tissue sections, where some } types of cells are expected to label while others are not. } } 2. Could somebody give me directions how to subscribe to HistoNet or any } other histochemical or immunohistochemical list? } } Thank you for your answers, } } Sarka Lhotak } Hamilton Cancer Center } Hamilton, Ontario, Canada } lhotaks-at-mcmaster.ca }
Thomas Geer, Marine Biological Laboratory, Woods Hole, MA, wrote: ================================================== ........ I am looking for coverslips or slides that will enable me to record the exact location of an object (calcite crystal) when viewed with a light microscope. I then would like to image the same crystal with an SEM, to view the morphology of the crystal. I have heard of slides or coverslips with grids that are labeled for just this purpose. I have not been able to find slides or coverslips for sale in any catalogs or on the web. I have also searched the archives with no luck. Perhaps someone has had similar experiences or knows just just what I am looking for. If so please let me know. ================================================= There are two "quick and dirty" (e.g. cheap and inexpensive) solutions to this kind of need, depending on particle size of the calcite crystals, as follows:
If less than about 5 um in size: ======================
a) Take a TEM index grid, however, our preference is for what we call our SPI Asbestos Index Grid because each square is precisely the same open area and with a very small standard deviation and with a few tiny (and I mean tiny) drops of a five minute epoxy, it is "glued" down to a glass cover slip . The generous "rim" area makes this quite easy. Another coverslip is placed over the whole set up in order to make the grid as flat as possible on the substrate cover slip. A "Teflon" coated part of a slide is what we have used in the event too much epoxy was used which could cause sticking if any other interface was used. The weights are placed on top of that Teflon coated part of a slide.
b) After the epoxy has cured, then disperse the calcite crystals preferably from some dilute non-interacting liquid suspension, preferably from an aerosol dispersion method (such as what is offered by Ernest F. Fullam, Inc. ) in order to minimize surface tension forces that could tend to drag the particles to the grid bars.
You now have exactly what you wanted, the particles mounted on a substrate that is clearly indexed, and something that can be put into either an LM or SEM and the same identical particle easily re-identified. And you have done this for almost no money at all!
If larger than 5 um: =============
Then the easiest way to do with is with the use of our own Tacky Dot Slides (TM). It is very easy to index each "dot" in the orthogonal matrix of "dots" and you can readily relocate the same dot in either the LM or SEM. And while the slides do cost "something" we are still talking about very low cost.
Information about the products mentioned in this posting can be found on our website given below. Both approaches easily lend themselves to quantitative microscopy. As a bonus benefit, if you weigh the Tacky Dot Slide before and after particles are attached, you can also do particle density calculations, something not otherwise easily done.
Disclaimer: SPI Supplies offers both the asbestos TEM grid product and the Tacky Dot Slide products and would have a vested interest in seeing more people adopting their use for this kind of sample preparation requirement. We have no commercial interest in the promotion of the Fullam product, it is just that our own in-house experience has shown us that it is the best available.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I recently obtained a Letiz Orthoplan scope from a biomedical colleague. I intend to use it for micropaleontological research, specifically, the identification of the siliceous skeletons of diatoms.
The scope is currently configured with a dark-field condenser and arc lamp for flourescence microscopy. I plan to reconfigure the scope for bright-field DIC and/or phase contrast work, as well as simple bright-field illumination.
The scope came with no documentation. As I prepare to reconfigure the scope with Leitz parts, I need to learn what lenses, condensers, and other components were once available. I wish to inquire if any listserv subscribers know where such a 20-year-old catalog might be found.
Once I have a better idea of what components I need, I will be interested in effecting an exchange of the fluoresence parts that I don't need (e.g., arc lamp power supply, lamp casing, two fluorescence objectives) for parts that will be useful for my intended use of the microscope.
I'll be looking for local advice (Seattle-Tacoma area) on the use of the scope and the Orthomat photographic system that came with it, but I'd also be grateful for any leads on documentation for this Orthoplan scope. I'd be willing to pay reasonable photocopy costs for manuals and catalogs.
For the next couple of days, please respond off the listserv, since I have very recently subscribed and may not yet be receiving all the listserv messages.
Hello appreciated microscopist. I am intested in entering in contac with some group that makes electronic Microscopy of cerebral tumors with purposes of diagnostic. I work in a service of Microscopy and I=A8m would want to put to disposition of hospitals this service.
} Does anyone have any suggestions for mounting ultrathin sections on } to carbon-coated, formar slotted grids? } TIA } } Hank Adams } EML } New Mexico State University }
First use a regular slotted grid(without formvar) and gently place it over the sections you want. By water tension the sections will cling to the grid. Then place slot of the regular grid over the slot of the formvar coated grid, and using a piece of filter paper, wick the water away. The sections will now be on your formvar grid. It takes two sets of forceps, preferably the self-closing kind. As for the regular grid(withour formvar) I also like to use the thicker Snaptek(sp?) kind. Good Luck!
Paula Webster Moore Bowman Gray School of Medicine Winston-Salem, N.C. pmoore-at-bgsm.edu
We have looked at a variety of powders ( including ceramic powders) using similar techniques to the one you described. Keep in mind that a lot of the agglomeration that you see is probably occurring while the drop of the dispersion is drying on the grid and therefore long times in the ultrasonic will not help. We often found that if the particles are not sintered together, then a few minutes of sonication will do. Other points to keep in mind are:
Dilute your dispersion as much as possible( you should barely notice a change in the turbidity of the liquid). This will minimize agglomeration during drying. Generally, I do not wait for the liquid to completely dry on the grid. Instead, after a few seconds ( 10-20 sec) , I remove the excess liquid using the corner of a filter paper or tissue.
If this does not work you could try spraying the dispersion onto the grids. Kits for doing this are available from various EM suppliers and they will also help minimize agglomeration during drying.
Another way is to mount your carbon coated grids onto a rotating plate ( I have improvised with a Dremel tool) so that when you place a drop on the grid it will spread out and minimize agglomeration.
Dear Colleagues: This is an old question but in an ever changing market: with a budget of $8,000 to $15,000, which make or model of dye sublimation printer gives the highest printing resolution? We will primarily use it for TEM B/W photos. Any information is greatly appreciated.
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Reply to: RE} } TEM: slotted grids
Dear Hank, If your grids are already formvar and carbon coated you can pick up the = sections two ways. One way is to just come down on top of the floating = sections gently and then bring them to the side of the boat removing = excess water using the surface tension to pull water into the boat while = moving the grid over the edge of the boat. The other is to go underneath = the sections (immerse the whole grid into the water) and bring it up = under the sections while keeping them in place using an eyelash. Then = lift the grid with sections out of the boat and carefully remove excess = water with filter paper. Linda Chicoine Center for Cell Imaging Yale University New Haven, CT 06520
--------------------------------------
} Does anyone have any suggestions for mounting ultrathin sections on } to carbon-coated, formar slotted grids? } TIA } } Hank Adams } EML } New Mexico State University }
First use a regular slotted grid(without formvar) and gently place it over the sections you want. By water tension the sections will cling to the grid. Then place slot of the regular grid over the slot of the formvar coated grid, and using a piece of filter paper, wick the water away. The sections will now be on your formvar grid. It takes two sets of forceps, preferably the self-closing kind. As for the regular grid(withour formvar) I also like to use the thicker Snaptek(sp?) kind. Good Luck!
Paula Webster Moore Bowman Gray School of Medicine Winston-Salem, N.C. pmoore-at-bgsm.edu
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I have been trying to use an apoptag staining kit to label cells in positive control tissue embedded in EM resins. I have attempted this procedure on both Epon and LR white with and without osmium, in semithin and ultrathin sections. Instead of the anti-digoxigenin-peroxidase-DAB reaction that we have successfully used on the histological level in parafin, I am substituting an anti-digoxigenin gold-cojugated label.
**The problem is that my staining labels ALL of the nuclei in the section instead of just the apoptotic ones. Has anyone else out there tried staining for apoptosis in resin embedded tissue or heard of successful attempts? ANY possible explanations for my results?
I also recently tried using a biotinylated apoptag in situ end-labeling kit from another company and had similar results.
Thanks for any help that you can offer.
Gregg Sobocinski Parke-Davis Pharmaceutical Research Division Ann Arbor, Michigan USA Sobocig-at-aa.wl.com
*** And I'm sure the lawyers would be happy if I include: The views expressed above are my individual opinions and do not represent the views or policies of Parke-Davis.
Today I am contacting the list as a representative for our group the Food Structure and Functionality Forum. After some discussion among ourselves, we have agreed to reply to this group about our experience.
As you may remember, the very well respected and excellent quality journal Food Microstructure (which later became Food Structure) grew out of the Johari meetings and the associated Food Microstructure Sessions. It was very unfortunate that, after a long and prosperous life, the Food Microstructure Sessions were not well attended, and the Food Microstructure journal was published sporadically, then, not at all. The Food Structure Researchers dearly missed having the Meeting and the Journal as places where they could present their work. Certain hard-working individuals tried to keep the food structure folks together, but it was difficult without a place to meet. Food journals were started in Europe and in North America, but none were completely dedicated to Structural studies.
The people at FAMS (Foundation for Advances in Medicine and Science) kindly provided us with a place to meet by agreeing to include a Food Structure and Functionality Session at their SCANNING 94 meeting in Charleston. The "Food session" has been a regular session at the SCANNING meetings ever since. The next year, the Food people met again at our session at SCANNING95 in Monterey, and FAMS helped and supported us to organize the Food Structure and Functionality Forum, and we developed a mandate. Since then, the Forum has been trying to find other food structure colleagues worldwide, compiling a mailing list. The Forum is still under development right now.
The latest development has been recently. We have just found out that this year's special issue of the Journal Scanning is being dedicated to Food Structure and Functionality papers, especially the ones presented in Baltimore at SCANNING 98.
Also, this year is the first year for another Food related session to be presented - Applications of Microscopy in Agriculture.
Anyone wanting further information about the sessions or the Forum can contact me or the FAMS website (http://www.scanning-fams.org).
Thanks for your interest and the opportunity to tell you about our experience.
Paula Allan-Wojtas Agricutlure and Agri-Food Canada Kentville, Nova Scotia Canada B4N 1J5
I apolgize to the list but this is in answer to Yuhui Xu's request for Dye-sub info. After 4 failed appemts to find a valid address from him I've resorted to replying back to the list.
------- Forwarded Message Follows -------
To: yuhui xu {-at-EYE.DFCI.HARVARD.EDU:Yuhui=Xu%RES-at-DFCI}
This is very similar to the question I asked a couple of weeks ago. The "Best resolution" is not really a good question. I think what you really want is "the best image quality for publication", since the printers in this range all seem to be 300DPI (The Epson Ink Jets range from 720 - 1440 DPI at $250 - $420, and the $420 Photo Stylus at 720DPI gives the best images of the group).
The simple answer is the Fuji Pictography at ~$13,500. It is NOT a dye-sub printer it is a 'dry' silver photography printer (but it needs to be fed water once a week). The next step down is the Codonics NP1660 (fully networked) at ~$12,500 (Or the Kodak 86xx? at a little less). The next step down is the Tektronics 650 at ~$9,000 (networked) but majority of the respondants thought the Condonics/Kodak had better images.
If you would like specific information comparing these printers I'm in the process of compiling the replies to my questions comparing the printers and I can forward a copy along to you.
Good luck.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
I would direct you to many publications by M Thiry who has extensively examined the use of transferase-immunogold techniques in several embedding resins. A recent article is found in Micros Res and Techniques 31:4-21, 1995, and he states that these techniques " are compatible with various fixation and embedding conditions". (Epon, Lowicryl K4M or LR White) Take this for what it's worth Marge
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
To All Our Friends, Because of severe weather conditions in northern New England, Ladd Research Industries has temporarily lost it's power and telephone service. We apologize for any inconvenience this may have caused. Hopefully we will reopen for business tomorrow, January 13. In an emergency, we may be reached at (802) 878-8074. Sincerely, Charles R. Duvic Chief Chemist Ladd Research Industries
Kovex Corporation, a rapidly growing high-tech manufacturer of surface inspection equipment specializing in the development of state of the art microscopy techniques, is looking for an engineering manager. See our web page for more information: www.kovexcorp.com
E.A. Fischione Instruments, Inc. is seeking a highly motivated individual for the position of Sales and Marketing Manager. Duties will include the maintenance of current and potential customer databases; trade show coordination; maintenance of World Wide Web site; coordination of pricing strategy; monitoring product performance with existing customers; generation of press releases for both product and company news; establishment of advertisements in trade publications; continuous updating of advertising literature; coordination of activities with foreign distributors and the establishment of new distributors; obtaining field input for the development of future products; coordination of direct mail campaigns; generation of sales projections, reports, and budgetary information.
The candidate should possess a B.S., M.S., or Ph.D. in materials science/engineering or physics and a Masters in Business Administration (MBA) with a focus on sales and marketing activities. As the Sales and Marketing Manager, a great deal of activity will focus on interactions with both existing and potential customers. The position requires excellent verbal and written communication skills and a willingness to do extensive travel.
E.A. Fischione Instruments, Inc. specializes in TEM specimen preparation instrumentation and TEM specimen holders. The corporate headquarters is in Export, Pennsylvania, approximately 23 miles east of Pittsburgh. For more information, contact our web site at www.fischione.com.
E.A. Fischione Instruments, Inc. is an equal opportunity employer.
Resumes should be sent to:
E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 USA
Hi, I was wondering if someone could help me with small definitions of what Scanning Transmission X-ray Microscopy and X-ray Imaging Microscopy are. Thanks. Scott
I would like to personally invite you to Arizona State University and Molecular Imaging's "HANDS ON" workshop on High Resolution In-Situ Scanning Probe Microscopy to be held in Phoenix, AZ; Feb. 4-11, 1998. http://green.la.asu.edu/workshop
Where: Arizona State University, Tempe, AZ; and Molecular Imaging Corporation, Phoenix, AZ
Two focused sessions: Extended time to run your samples can be arranged. (If you plan on bringing samples please contact session head to assure adequacy of sample preparation) - Electrochemical SPM (Feb 4-6), Poster/cocktails Faculty club Feb 5 Session head: Dr. Judy Zhu mailto:Judy-at-molec.com - Biological SPM (Feb9-11), Poster/cocktails Faculty club Feb 10 Session head: Dr. Jun Li mailto:junli-at-molec.com
A general introduction to in situ SPM will start each session. Included will be an overview of SPM theory, and an introduction to sample preparation techniques for imaging under controlled atmosphere, in fluids, under temperature or electrochemical control, and or high resolution imaging and force measurement techniques (MAC Mode) ----------------------------------------------- Workshop Goals: The goal of this workshop is to provide a forum for presentations, hands on training (on attendee samples), and discussion of recent developments in research, instrumentation, current applications and future directions of high-resolution in-situ STM and AFM. Some areas of particular interest are: - Observing molecular conformational change in situ - Magnetic AC imaging (MAC Mode) - MAC Force measurements - Corrosion studies - Battery studies - Biomolecular imaging and force/friction/electrical data analysis (functionalized tips) - In situ biomolecular research (gene therapy research, drug discovery, drug delivery, diagnostic sensors) - Micro manipulation (protein folding force, prolymer characterization) - Environmental control - Temperature control - In-situ imaging, Sample preparation, SPM techniques (Tips and Tricks) - New developments in instrumentation. ----------------------------------------------- Session I: ELECTROCHEMISTRY AND IN-SITU IMAGING This session focuses on SPM issues important to corrosion, catalysis, and advanced battery research. - Investigating potential dependant effects of surfaces and surface adsorbates - Electrochemical theory and techniques in corrosion, Catalysis, and battery technology - Real time monitoring of surface morphology } due to potential or } electrolyte induced corrosion or } during potential cycling and in presents of reactive species - Investigating effects of corrosion inhibitors - Preparing and working with air and moisture sensitive samples - Monitoring the evolution of surface
Techniques Demonstrated: Controlled environment, controlled temperature, in-situ, and electrochemical STM and AFM
Research areas: General electrochemistry, corrosion, catalysis, and battery research -----------------------------------------------
Session II: BIOLOGY AND HIGH RESOLUTION IN SITU IMAGING AND CHARACTERIZATION This session focuses on applications of high resolution SPM for studying biological samples both in situ and ex situ. - Applications of SPM in biological research - Preparing and working with biological samples - Low force biological imaging using magnetic AC - In situ imaging of bio samples under environmental and temperature control - Force measurement and sample characterization
Techniques Demonstrated: Contact AFM, controlled temperature, in-situ MAC mode, MAC mode with temperature control
Research areas: Molecular and cell biology, structural biology, drug discovery and drug delivery. -----------------------------------------------
Speakers and Tutors: - Prof. Stuart Lindsay, ASU - Dr. Daphna Yaniv, MI - Dr. Judy Zhu, ASU/MI - Dr. James Hudson, ASU/MI - Dr. Jun Li, ASU/MI - Dr. Tainwei Jing, Cofounder of MI
Who should attend: Researchers and microscopists from beginner to expert. Attendees will be grouped both by interest and ability. Groups will be 3-5 and each group will have a dedicated system for hands on experience.
Cost: $575 for one workshop session, $975 for both workshops sessions. (Includes lunches, and evening program, and workshop materials)
TO SIGN UP or more information please go to http://green.la.asu.edu/workshop
For more information, e-mail: - Elena Odenheim mailto:elena-at-molec.com or - Prof. "Stuart Lindsay" mailto:stuart.lindsay-at-asu.edu
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
To all SEM microscopists who have been waiting for scanning probe microscopy to deliver unique capabilities. "High resolution in situ imaging and sample characterization under 37 C controlled temperature".
I would like to personally invite you to Arizona State University and Molecular Imaging's "HANDS ON" workshop on High Resolution In-Situ Scanning Probe Microscopy to be held in Phoenix, AZ; Feb. 4-11, 1998. http://green.la.asu.edu/workshop
Where: Arizona State University, Tempe, AZ; and Molecular Imaging Corporation, Phoenix, AZ
Two focused sessions: Extended time to run your samples can be arranged. (If you plan on bringing samples please contact session head to assure adequacy of sample preparation) - Electrochemical SPM (Feb 4-6), Poster/cocktails Faculty club Feb 5 Session head: Dr. Judy Zhu mailto:Judy-at-molec.com - Biological SPM (Feb9-11), Poster/cocktails Faculty club Feb 10 Session head: Dr. Jun Li mailto:junli-at-molec.com
A general introduction to in situ SPM will start each session. Included will be an overview of SPM theory, and an introduction to sample preparation techniques for imaging under controlled atmosphere, in fluids, under temperature or electrochemical control, and or high resolution imaging and force measurement techniques (MAC Mode) ----------------------------------------------- Workshop Goals: The goal of this workshop is to provide a forum for presentations, hands on training (on attendee samples), and discussion of recent developments in research, instrumentation, current applications and future directions of high-resolution in-situ STM and AFM. Some areas of particular interest are: - Observing molecular conformational change in situ - Magnetic AC imaging (MAC Mode) - MAC Force measurements - Corrosion studies - Battery studies - Biomolecular imaging and force/friction/electrical data analysis (functionalized tips) - In situ biomolecular research (gene therapy research, drug discovery, drug delivery, diagnostic sensors) - Micro manipulation (protein folding force, polymer characterization) - Environmental control - Temperature control - In-situ imaging, Sample preparation, SPM techniques (Tips and Tricks) - New developments in instrumentation. ----------------------------------------------- Session I: ELECTROCHEMISTRY AND IN-SITU IMAGING This session focuses on SPM issues important to corrosion, catalysis, and advanced battery research. - Investigating potential dependant effects of surfaces and surface adsorbates - Electrochemical theory and techniques in corrosion, Catalysis, and battery technology - Real time monitoring of surface morphology } due to potential or } electrolyte induced corrosion or } during potential cycling and in presents of reactive species - Investigating effects of corrosion inhibitors - Preparing and working with air and moisture sensitive samples - Monitoring the evolution of surface
Techniques Demonstrated: Controlled environment, controlled temperature, in-situ, and electrochemical STM and AFM
Research areas: General electrochemistry, corrosion, catalysis, and battery research -----------------------------------------------
Session II: BIOLOGY AND HIGH RESOLUTION IN SITU IMAGING AND CHARACTERIZATION This session focuses on applications of high resolution SPM for studying biological samples both in situ and ex situ. - Applications of SPM in biological research - Preparing and working with biological samples - Low force biological imaging using magnetic AC - In situ imaging of bio samples under environmental and temperature control - Force measurement and sample characterization
Techniques Demonstrated: Contact AFM, controlled temperature, in-situ MAC mode, MAC mode with temperature control
Research areas: Molecular and cell biology, structural biology, drug discovery and drug delivery. -----------------------------------------------
Speakers and Tutors: - Prof. Stuart Lindsay, ASU - Dr. Daphna Yaniv, MI - Dr. Judy Zhu, ASU/MI - Dr. James Hudson, ASU/MI - Dr. Jun Li, ASU/MI - Dr. Tainwei Jing, Cofounder of MI
Who should attend: Researchers and microscopists from beginner to expert. Attendees will be grouped both by interest and ability. Groups will be 3-5 and each group will have a dedicated system for hands on experience.
Cost: $575 for one workshop session, $975 for both workshops sessions. (Includes lunches, and evening program, and workshop materials)
TO SIGN UP or more information please go to http://green.la.asu.edu/workshop
For more information, e-mail: - Elena Odenheim mailto:elena-at-molec.com or - Prof. "Stuart Lindsay" mailto:stuart.lindsay-at-asu.edu
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Meeting announcement - Focus on Microscopy 1998 Full details follow - best viewed in a monospaced font.
For a classier view see our web page created by Pal Fekete http://www.physics.usyd.edu.au/physopt/fm98
Online registration will be available on the web page soon but you can also get a form (and any further details) by email from focus98-at-emu.usyd.edu.au
Focus on Microscopy 1998 11th International Conference on 3D Image Processing in Microscopy 10th International Conference on Confocal Microscopy
April 14th-17th, 1998
University of Sydney, New South Wales, Australia
Australian Key Centre for Microscopy and Microanalysis Royal Microscopical Society (UK) Image Analysis Society of Australia
International Committee Prof. Colin Sheppard, University of Sydney Prof. G.J. Brakenhoff, University of Amsterdam Prof. Tony Wilson, University of Oxford Dr. Vyvyan Howard, University of Liverpool Dr. Andres Kriete, Liebig University, Giessen Prof. P-C. Cheng, SUNY at Buffalo Prof. Alan Boyde, University of London Dr. Guy Cox, University of Sydney Prof. S. Kawata, Osaka University
Organising Committee Prof. Colin Sheppard, University of Sydney Dr. Guy Cox, University of Sydney Ms. Carol Cogswell, University of Sydney Dr. Pal Fekete, University of Sydney Dr. Min Gu, Victoria University Dr. Allan Jones, University of Sydney Ms. Eleanor Kable, University of Sydney
Introducing Sydney
Sydney, Australia's largest city, is also one of the world's most beautiful cities, built around the spectacular natural harbour which provided the site for the first European settlement of the Australian continent. It prides itself on its cultural diversity, offering a rich mix of European, Asian and indigenous Australian experiences alongside the uniquely Australian culture which has developed in the 200 years since the First Fleet landed in Sydney Cove.
The University of Sydney is the oldest in the country, established as part of the great English 19th century tradition of liberal enlightenment but unashamedly modelled architecturally on the Oxbridge pattern. It has retained both its prestige and its central location although its site has expanded greatly over the years, now accommodating more than 20,000 students. The Australian Key Centre for Microscopy and Microanalysis (AKCMM) at the University of Sydney, which is hosting the Conference this year, is the largest centre for microscopy in the Southern Hemisphere, offering a wide range of optical and electron microscope facilities both to the University and the wider community.
April is autumn (fall) in Sydney. The weather will be mild average temperature for the month is 19 C (68 F). Sydney has both a lot of sun and a high rainfall - rain can fall in any month so bring a waterproof. The ocean will be warm and very pleasant for swimming and surfing.
Scientific Programme - 15-17 April
The scientific programme will consist of poster and spoken sessions. Posters (1m x 1m) and contributed talks (15min) are invited on any of the Conference topics:
* Advances in confocal microscopy * Applications of confocal microscopy * 3-dimensional optical imaging * 3-D techniques in electron microscopy * Other 3D imaging techniques * Novel techniques in microscopy * Near-field microscopy * Multiple-photon microscopy * Multiple-dimensional image processing * Applications of image analysis
Short Courses & Workshops - 14th April
The following half-day short courses and workshops will be offered, subject to both maximum and minimum numbers of participants. All are led by internationally recognized experts in their respective fields. The cost is $75 (Aust) per half-day course.
Morning * Multiphoton microscopy * Introductory confocal microscopy * Deconvolution of 3D images * Stereology
Afternoon * Introduction to image processing * Advanced confocal microscopy * Introduction to digital imaging * 3D image processing & visualization
Social Programme
These events are included in the cost of full and accompanying member registration. A limited number of additional tickets will be available.
* Tuesday 14th April, 6pm. Welcome reception, exhibition area. Drinks and simple snacks - an opportunity to meet old friends and to get to know fellow delegates before the start of the formal business of the conference. * Thursday 16th April, 7pm. Conference Barbecue Dinner. An informal evening on an island in one of the most beautiful parts of Sydney Harbour.
Morning coffee, a light lunch, and afternoon tea are provided each day. (For workshop registrants only on the 14th).
Abstracts
Extended abstracts, up to one A4 page in length, will be published in a conference volume issued free to all delegates. Additional copies will be available for sale. Micrographs and other illustrations (monochrome only) are welcomed but must fit within the one page.
Manuscripts will be edited for format only. To simplify the editors' task please follow these simple guidelines: * Title in upper and lower case. * Authors' names with initials first, presenting author in upper case. * Full address and affiliation of all authors. * Text in a 12pt font. * References cited by name and date, not number. * References at end - do not include titles. All authors (initials first), then year, then journal citation.
All text must be submitted electronically, either as plain text, RTF or in the format of a PC or Macintosh word processor. MS Word, Word Perfect, Wordstar, MS Works, Claris Works, Write are all on site and most other formats can be converted. Postscript files and TeX are not acceptable. Illustrations should not be included within the word processor file; they should be submitted either as separate files in any common format (not postscript or eps) or as hard copy.
Send files : * by email to focus98-at-emu.usyd.edu.au * by anonymous ftp to ftp.emu.usyd.edu.au (directory \focus98) ftp://ftp.emu.usyd.edu.au/focus98) * on a floppy disk to the conference address, below.
Abstracts must be received by 31st January 1998 if they are to appear in the published volume.
Conference Details
Conference sessions and a comprehensive manufacturers' exhibition will take place in the Wentworth Building, levels 4 & 5. Some workshops will be held in the AKCMM, Madsen Building. A footbridge across City Road provides quick access between these buildings.
A discounted early registration fee applies to all registrations received and paid by 31st January 1997. All prices given are in Australian dollars - one Australian dollar is approximately 70c US.
Early Regular Full registration $ 425 $ 475 Student registration $ 250 $ 300 Day registration $ 175 $ 175 Accompanying person $ 175 $ 175
Full registration covers conference volume, admission to all scientific sessions, welcome reception, conference dinner, morning and afternoon tea, and lunch.
Student registration includes all the above except the conference dinner.
Day registration covers conference volume, admission to scientific sessions, morning and afternoon tea, and lunch on any one day.
Accompanying person's registration includes welcome reception, two half-day tours and the conference dinner
Accommodation is available at St John's College, on the University campus, for $60 per night including breakfast (single room, shared bathroom). For those who prefer a hotel, Camperdown Travelodge is $125 per night (single occupancy) or $135 (dual occupancy) including breakfast. These are specially discounted rates - to obtain them you must book through the conference. The taxi fare from the airport to Wentworth Building, St. John's College or the Travelodge is about $10.
Focus on Microscopy '98, Australian Key Centre for Microscopy and Microanalysis, F09, University of Sydney, NSW 2006, Australia.
Discounted registration for Focus on Microscopy 1998 -11th International Conference on 3D Image Processing in Microscopy -10th International Conference on Confocal Microscopy finishes on 31st January !
You have three more weeks to avoid paying full price.
Check out our web page: http://www.physics.usyd.edu.au/physopt/fm98
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear colleagues, I'm trying to stain the mesodermal cells of sea urchin embryo with = Ruthenium Red, in block. Does somebody now the way to destroy the tight = junctions of epitelial cells to to make it easier for RuRed to enter the = blastocoel. I'll appriciate any help or suggestions, thank you in = advance, Elia
Elia Beniash Structural Biology Weizmann Institute of Science Rehovot, Israel
I need a gold standard to calibrate an EDS analytical system attatched to a TEM.
I've contacted several suppliers but none of them have anything with gold (pure or in a compound).
Does anyone know where or how I can get a thin film standard of pure gold or of a compound including gold ?
Thanks in advance.
Isabel Nogueira Instituto Superior T=E9cnico Departamento de Materiais Avenida Rovisco Pais, 1096 Lisboa Codex PORTUGAL telef: 351-1-8418124/0 fax: 351-1-8418120 e-mail: isabeln-at-alfa.ist.utl.pt
As many of you know, northern New England was hit with the "ice storm of the century" last week. Ladd Research and a good chunk of the rest of this area lost power, heat and telephone systems on Thursday. As of now (Tues, 8:30 AM EST) we just got our electricity back, but our phone system is going in and out. Hopefully by the time this message is posted they will have finished the repairs and we will be fully operational. 1-800-451-3406 should get you through, if not we have forwarded everything through the fax number that doesn't go through the switchboard so you can call 802-878-8074. Once everything is back that will once again be the fax number. If you can't get thru by phone please either keep trying or e-mail us. I have over 150 messages to go through but we will reply to everyone as soon as possible. To those who have been trying to fax please try again , that should be operational now. For those who have orders placed with us, we will be shipping today.
Thank you for being understanding during this disruption.
I am gathering information on Microscopy Facilities which rent time to industry. I am interested in facilities which have the following capabilities - e.g. High resolution TEM and STEM, EELS and Image Filtering, High Angle Annular Dark Field. I would appreciate any help gathering this info.
The Analytical and Materials Characterization Laboratory at Polaroid Corp. has a need for a microscopist with a background in Material Science to join our Microscopy and Surface Analysis Group.
Strong problem solving, leadership, interpersonal, documentation and communication skills are a prerequisite.
Skills in Atomic Force Microscopy and Light Microscopy as well as a variety of sample preparation techniques i.e. microtomy are essential.
Strong computer skills with application to digital imaging, remote imaging and image analysis are desirable.
Experience in scanning electron microscopy, transmission electron microscopy, confocal microscopy, x-ray photoelectron spectroscopy (XPS) and x-ray diffraction is desirable.
A background in fracture mechanics and/or process microscopy is also desirable.
This position requires a Ph.D. in materials science, physics, chemistry, or engineering or a M.S. with equivalent training. Industrial experience is a plus.
Supervisor : Lynne Garone, e. mail: GaroneL-at-Polaroid.com
Location: W4-1D 1265 Main St. Waltham, Ma. 02254 (781) 386-1446 Fax: (781) 396-0378
Many thanks to those who responded to my layman's question on Dye Sub printer. From what I understood from these responses, with a budget of under $15,000, one has the choices of the following dye sub printers: Kodak 8650, Mitrubishi S3600-40U, Tektronix 560, or Codonics NP1600 ( I was told by a local salesman that this line of products have been discotinued. But apparently they are still available from some dealers). The Fuji Pictography is said to work by a different mechanism. Here I have two more questions:
1) I learned from those responses that all dye sub printers print at 300 dpi resolution. But the print out pictures look much smoother than those produced by laser or Inkjet printers printed at much higher resolution. Is this due to the difference in printing mechanisms? My understanding is that the space between the 300 dots must has been "filled" by "continuous tone" with the same information from the dots next by.
2) If the print out is only at 300 dpi, does that mean that we only need a negative scanner that reads and inputs images at equal or slightly higher than 300dpi?
Sorry for asking again these very basic quuestions. Regards,
I have seen your message over the server. Here at Electron Microscopy Sciences we can offer you a Thin film standard with pure gold. This is a standard item of ours. Please see page 90 of our catalog or at our website at www.emsdiasum.com. Please let us know if we may be of further assistance to you.
} ... } Many thanks to those who responded to my layman's question on Dye Sub } printer. ... } Here I have two more questions: } } 1) I learned from those responses that all dye sub printers print at 300 dpi } resolution. But the print out pictures look much smoother than those produced } by laser or Inkjet printers printed at much higher resolution. Is this due to } the difference in printing mechanisms? My understanding is that the space } between the 300 dots must has been "filled" by "continuous tone" with the } same information from the dots next by.
The different "mechanism" is actually a different method of mixing cyan, magenta, yellow ( and possibly black) to create the printable colors. Lasers and inkjets use the "dithering" method and need to put C,M,Y & K dots very close together in order to produce an illusion of the resultant color ("dithering" is the newspaper method). Dye-subs on the other hand can actually "mix" C,M,Y & K to produce the color ... ie, no dithering is needed. The result is at 300dpi dye-subs produce much smoother colors than dithering, yet the new high-resolution Epsons (and HPs) do a *very* good job, and the dithering is almost un-noticeable. Newer ink jets have recently introduces 6color printing which add light cyan and magenta to the dithering process ... this improves printing considerably ... and now you have a high quality printer for less than $1000. For the sake of $$ per page, you might want to consider this an addition printer to buy.
} 2) If the print out is only at 300 dpi, does that mean that we only need a } negative scanner that reads and inputs images at equal or slightly higher } than 300dpi?.
This would depend on the final size of your printed page. For example, if you wanted an 8by10 printed from a 4by5 negative, then you'd want to scan it at 600dpi.
What is nice about dye-sub printing is that you don't have to be a mathematician to calculate "lines-per-inch" which accounts for the size of the dithering matrix. Yet, dithering printer manufacturers will suggest the "ideal" dpi for photographic color printing ... for example, Epson suggests 240dpi for its 720dpi printers. Since a dye-sub doesn't use a dithering matrix the printing resolution of 300dpi is straight-forward.
... hope this helps :o) cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility at the University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
I draw your attention to the fact that the Swiss Society for Optics and Microscopy has changed his Web site's address. It was previously www.sgoem.ch - it is now: www.ssom.ch
Thanks for you attention, Best regards,
Robin Schaublin
-- ------------------------------------------------------------- Robin E. Schaeublin Centre de Recherches en Physique des Plasmas - EPFL Fusion Technology - Materials Group CH - 5232 Villigen - PSI, SWITZERLAND Tel : + 41 56 310 40 82 and + 41 21 693 65 69 Fax : + 41 56 310 45 29 and + 41 21 693 37 51 -------------------------------------------------------------
Dear Tom, The Leica rep. for our area is Bartels and Stout, in Issaquah, (425)453-1705. They may be able to help you with manuals and technical assistance. You might even be able to bring it in for a tutorial. The Orthomat camera system is very reliable and easy to use. If all else fails, I could give you a lesson. If you have extra parts, our lab might be interested in them.
My mail server had problems with your email address yesterday, so I'm sending a cc: to the list today.
Regards, Glen MacDonald Virginia Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu *---------------------------------------------------------------------* The box said "Requires Windows 95 or better.", so I bought a Macintosh. *---------------------------------------------------------------------*
On Sun, 11 Jan 1998 16:09:06 EST TomDeVrie-at-aol.com (Tom DeVrie) wrote:
} } The scope came with no documentation. As I prepare to reconfigure the } scope } with Leitz parts, I need to learn what lenses, condensers, and other } components were once available. I wish to inquire if any listserv } subscribers } know where such a 20-year-old catalog might be found.
I'm having a strange problem with our Philips XL30. And, I was wondering if anyone has had any similar difficulties, or if anyone can offer any suggestions.
The problem is with either saving to .tif file or restoring from it. The databar is restored unreadably. The image is restored fine, and you can tell that the letters of the graphic are restored, but the background is then distorted so that the databar section is unreadable. The databar is fine when the image is sent to the screen, videoprinter, or camera. I have also tried to pull up the .tif file using other programs with the same results.
We are nearly back all the way. 1-800-451-3406 will get you through, as will 1-802-878-6711 for those not in the United States. Our fax, 1-802-878-8074 is also working again.
When they put the telephone poles back up outside our plant and turned the power back on our phone system cpu was fried. We have rigged up a quick fix, but only have half of our usual lines for now so you may get a busy signal or no answer if the hunting lines are not set the way we think they are, but rest assured we are operating.
Thank you again for your understanding in this matter.
There is some sort of minor software problem with our Edax computer. A few times a day, during heavy use, the computer will declare a general protection error. The only way out of this error is to close all programs and exit windows and then restart. After restart nothing appears to be strange, until several hours later another general protection error occurs. I have been looking for a pattern, but have been unsuccessful to date.
Recently there have been some very good suggestions posted regarding posture and positioning of the body during microscopy to minimize fatigue and injury. However, no one has mentioned the design of microscopes. We seem to be resigned to adjusting our bodies to fit this awkward instrument. I would like to encourage microscope manufacturers to modify microscope designs to fit our bodies. With our backs and necks straight we should be able to look directly, or down a few degrees, into the eyepieces. The principle operating controls, focus and stage, should be at a level near our waist so that our arms are bent at the elbows 90 degrees or slightly more. In the case of dissecting microscopes the stage level should be at this waist height. There are many diagrams circulating showing how to best position the body while using a computer. These also pertain to using other equipment such as microscopes. I have already discussed these issues with some Nikon Product Managers and encourage you to gently nudge manufacturers in this direction before that soreness in the shoulders becomes a chronic pain. Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
} ... } Is there a difference between histogram equalization and histogram stretching? } ...
The two terms are most commonly used interchangably, but there is a difference. "Stretching" would simply expand the distribution of gray levels such that the brightest is 0 and the darkest is 255 ... whereas, "equalization" is a bit more difficult to describe. John Russ ("Computer Assisted Microscopy", Plenum, 1991) puts it this way ...
"With this method, an equal number of pixels in the final image have each available shade of gray. The shape of the transfer function can be straightforwardly determined from the brightness histogram of the original image. The method expands the contrast in regions of high gradients, so the details are much easier to see"
His figures go a long way to make this clearer ... suffice to say it is not a linear transfer function and dependent on the histogram gray level distribution with regard to how many pixels are in dark areas versus how many are in bright areas.
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility at the University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
I would suggest using an aluminum thin film on a copper grid for your energy calibration sample for the EDS sample. Bring your beam close to a grid bar and you will get sizable Cu Ka and Kb peaks. You will be able to adjust the energy scale quite easily and you will have a lower energy peak in the Al K (and a Cu -L peak for light element detectors.) that you can also check.
By having two peaks that you can measure the FWHM on, you can determine what the resolution of any peak in your system should be, e.g. Mn-Ka.
You can get both gold and aluminum thin films on copper grids from any EM supplier.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I need a gold standard to calibrate an EDS analytical system attatched to a TEM.
I've contacted several suppliers but none of them have anything with gold (pure or in a compound).
Does anyone know where or how I can get a thin film standard of pure gold or of a compound including gold ?
Thanks in advance.
Isabel Nogueira Instituto Superior Ticnico Departamento de Materiais Avenida Rovisco Pais, 1096 Lisboa Codex PORTUGAL telef: 351-1-8418124/0 fax: 351-1-8418120 e-mail: isabeln-at-alfa.ist.utl.pt
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Dear colleagues, I'm trying to stain the mesodermal cells of sea urchin embryo with Ruthenium Red, in block. Does somebody now the way to destroy the tight junctions of epitelial cells to to make it easier for RuRed to enter the blastocoel. I'll appriciate any help or suggestions, thank you in advance, Elia
Elia Beniash Structural Biology Weizmann Institute of Science Rehovot, Israel
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Meeting announcement - Focus on Microscopy 1998 Full details follow - best viewed in a monospaced font.
For a classier view see our web page created by Pal Fekete http://www.physics.usyd.edu.au/physopt/fm98
Online registration will be available on the web page soon but you can also get a form (and any further details) by email from focus98-at-emu.usyd.edu.au
Focus on Microscopy 1998 11th International Conference on 3D Image Processing in Microscopy 10th International Conference on Confocal Microscopy
April 14th-17th, 1998
University of Sydney, New South Wales, Australia
Australian Key Centre for Microscopy and Microanalysis Royal Microscopical Society (UK) Image Analysis Society of Australia
International Committee Prof. Colin Sheppard, University of Sydney Prof. G.J. Brakenhoff, University of Amsterdam Prof. Tony Wilson, University of Oxford Dr. Vyvyan Howard, University of Liverpool Dr. Andres Kriete, Liebig University, Giessen Prof. P-C. Cheng, SUNY at Buffalo Prof. Alan Boyde, University of London Dr. Guy Cox, University of Sydney Prof. S. Kawata, Osaka University
Organising Committee Prof. Colin Sheppard, University of Sydney Dr. Guy Cox, University of Sydney Ms. Carol Cogswell, University of Sydney Dr. Pal Fekete, University of Sydney Dr. Min Gu, Victoria University Dr. Allan Jones, University of Sydney Ms. Eleanor Kable, University of Sydney
Introducing Sydney
Sydney, Australia's largest city, is also one of the world's most beautiful cities, built around the spectacular natural harbour which provided the site for the first European settlement of the Australian continent. It prides itself on its cultural diversity, offering a rich mix of European, Asian and indigenous Australian experiences alongside the uniquely Australian culture which has developed in the 200 years since the First Fleet landed in Sydney Cove.
The University of Sydney is the oldest in the country, established as part of the great English 19th century tradition of liberal enlightenment but unashamedly modelled architecturally on the Oxbridge pattern. It has retained both its prestige and its central location although its site has expanded greatly over the years, now accommodating more than 20,000 students. The Australian Key Centre for Microscopy and Microanalysis (AKCMM) at the University of Sydney, which is hosting the Conference this year, is the largest centre for microscopy in the Southern Hemisphere, offering a wide range of optical and electron microscope facilities both to the University and the wider community.
April is autumn (fall) in Sydney. The weather will be mild average temperature for the month is 19 C (68 F). Sydney has both a lot of sun and a high rainfall - rain can fall in any month so bring a waterproof. The ocean will be warm and very pleasant for swimming and surfing.
Scientific Programme - 15-17 April
The scientific programme will consist of poster and spoken sessions. Posters (1m x 1m) and contributed talks (15min) are invited on any of the Conference topics:
* Advances in confocal microscopy * Applications of confocal microscopy * 3-dimensional optical imaging * 3-D techniques in electron microscopy * Other 3D imaging techniques * Novel techniques in microscopy * Near-field microscopy * Multiple-photon microscopy * Multiple-dimensional image processing * Applications of image analysis
Short Courses & Workshops - 14th April
The following half-day short courses and workshops will be offered, subject to both maximum and minimum numbers of participants. All are led by internationally recognized experts in their respective fields. The cost is $75 (Aust) per half-day course.
Morning * Multiphoton microscopy * Introductory confocal microscopy * Deconvolution of 3D images * Stereology
Afternoon * Introduction to image processing * Advanced confocal microscopy * Introduction to digital imaging * 3D image processing & visualization
Social Programme
These events are included in the cost of full and accompanying member registration. A limited number of additional tickets will be available.
* Tuesday 14th April, 6pm. Welcome reception, exhibition area. Drinks and simple snacks - an opportunity to meet old friends and to get to know fellow delegates before the start of the formal business of the conference. * Thursday 16th April, 7pm. Conference Barbecue Dinner. An informal evening on an island in one of the most beautiful parts of Sydney Harbour.
Morning coffee, a light lunch, and afternoon tea are provided each day. (For workshop registrants only on the 14th).
Abstracts
Extended abstracts, up to one A4 page in length, will be published in a conference volume issued free to all delegates. Additional copies will be available for sale. Micrographs and other illustrations (monochrome only) are welcomed but must fit within the one page.
Manuscripts will be edited for format only. To simplify the editors' task please follow these simple guidelines: * Title in upper and lower case. * Authors' names with initials first, presenting author in upper case. * Full address and affiliation of all authors. * Text in a 12pt font. * References cited by name and date, not number. * References at end - do not include titles. All authors (initials first), then year, then journal citation.
All text must be submitted electronically, either as plain text, RTF or in the format of a PC or Macintosh word processor. MS Word, Word Perfect, Wordstar, MS Works, Claris Works, Write are all on site and most other formats can be converted. Postscript files and TeX are not acceptable. Illustrations should not be included within the word processor file; they should be submitted either as separate files in any common format (not postscript or eps) or as hard copy.
Send files : * by email to focus98-at-emu.usyd.edu.au * by anonymous ftp to ftp.emu.usyd.edu.au (directory \focus98) ftp://ftp.emu.usyd.edu.au/focus98) * on a floppy disk to the conference address, below.
Abstracts must be received by 31st January 1998 if they are to appear in the published volume.
Conference Details
Conference sessions and a comprehensive manufacturers' exhibition will take place in the Wentworth Building, levels 4 & 5. Some workshops will be held in the AKCMM, Madsen Building. A footbridge across City Road provides quick access between these buildings.
A discounted early registration fee applies to all registrations received and paid by 31st January 1997. All prices given are in Australian dollars - one Australian dollar is approximately 70c US.
Early Regular Full registration $ 425 $ 475 Student registration $ 250 $ 300 Day registration $ 175 $ 175 Accompanying person $ 175 $ 175
Full registration covers conference volume, admission to all scientific sessions, welcome reception, conference dinner, morning and afternoon tea, and lunch.
Student registration includes all the above except the conference dinner.
Day registration covers conference volume, admission to scientific sessions, morning and afternoon tea, and lunch on any one day.
Accompanying person's registration includes welcome reception, two half-day tours and the conference dinner
Accommodation is available at St John's College, on the University campus, for $60 per night including breakfast (single room, shared bathroom). For those who prefer a hotel, Camperdown Travelodge is $125 per night (single occupancy) or $135 (dual occupancy) including breakfast. These are specially discounted rates - to obtain them you must book through the conference. The taxi fare from the airport to Wentworth Building, St. John's College or the Travelodge is about $10.
Focus on Microscopy '98, Australian Key Centre for Microscopy and Microanalysis, F09, University of Sydney, NSW 2006, Australia.
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REMINDER
Discounted registration for Focus on Microscopy 1998 -11th International Conference on 3D Image Processing in Microscopy -10th International Conference on Confocal Microscopy finishes on 31st January !
You have three more weeks to avoid paying full price.
Check out our web page: http://www.physics.usyd.edu.au/physopt/fm98
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E.A. Fischione Instruments, Inc. is seeking a highly motivated individual for the position of Sales and Marketing Manager. Duties will include the maintenance of current and potential customer databases; trade show coordination; maintenance of World Wide Web site; coordination of pricing strategy; monitoring product performance with existing customers; generation of press releases for both product and company news; establishment of advertisements in trade publications; continuous updating of advertising literature; coordination of activities with foreign distributors and the establishment of new distributors; obtaining field input for the development of future products; coordination of direct mail campaigns; generation of sales projections, reports, and budgetary information.
The candidate should possess a B.S., M.S., or Ph.D. in materials science/engineering or physics and a Masters in Business Administration (MBA) with a focus on sales and marketing activities. As the Sales and Marketing Manager, a great deal of activity will focus on interactions with both existing and potential customers. The position requires excellent verbal and written communication skills and a willingness to do extensive travel.
E.A. Fischione Instruments, Inc. specializes in TEM specimen preparation instrumentation and TEM specimen holders. The corporate headquarters is in Export, Pennsylvania, approximately 23 miles east of Pittsburgh. For more information, contact our web site at www.fischione.com.
E.A. Fischione Instruments, Inc. is an equal opportunity employer.
Resumes should be sent to:
E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 USA
I have run into similar problems on at least three occasions where the memory was (or was getting) flaky. Even though I ran several different memory diagnostics, the problem resisted a solution. Finally, trying some other memory SIMMs cleared the problem up. Memory is cheap enough now ($40 for 16 MB) that it is an inexpensive experiment. And you can always use more memory someplace these days.
On my second experience with this, I had people from Microsoft and elsewhere telling a colleague to reformat the hard drive and load a clean version of the software (and other drastic advice). I felt vindicated when it turned out to be simply the memory after having found that to be the solution the first time. Then I ran into it a third time, and that was on a Mac.
There may be other explanations as well. I have seen cooling fans fail and cause overheating in the processor. But that normally leads to a more severe and abrupt failure in the software.
Hope this helps.
At 09:47 AM 1/13/98 -0800, you wrote: } Hi, } } There is some sort of minor software problem with our Edax } computer. A few times a day, during heavy use, the computer } will declare a general protection error. The only way out of } this error is to close all programs and exit windows and then } restart. After restart nothing appears to be strange, until } several hours later another general protection error occurs. } I have been looking for a pattern, but have been unsuccessful } to date. } } Any and all suggestions are much appreciated. } } Thanks, } Jill Craig } UNBC ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello: } } Is there a difference between histogram equalization and histogram stretching? } } Owen } }
Yes. In histogram stretching, you are simply stretching the outliers of the pixel values to fit the dynamic range of the representation, and scaling everything inbetween accordingly. The *relative* position of the pixels intensity values will not change. With histogram equalization, you change the relative positions in order to distribute the values more evenly within the representation. Histogram equalization does not *necessarily* imply histogram stretch as well, though most implementations go ahead and do it.
Here's an example.
Consider a histogram that looks like this:
Many | | | # of pixels | * | *** | B****** | ******B****** Few | ***********B******* ------------------------------------ Dark Light
Brightness
Note that the column denoted with "B"s instead of asterisks is roughly in the middle of the distribution.
A histogram stretch will scale everything to go from the darkest possible representation to the brightest possible representation:
Many | | | # of pixels | * | * * * | B * * * * * * | * * * * * * B * * * * * * Few |* * * * * * * * * B * * * * * * * * ------------------------------------ Dark Light
Brightness
Note that the column denoted by "B"s is still roughly in the middle.
Histogram equalization recognizes that, by some statistical measures the greatest overall contrast enhancement (in terms, for instance, of information content) is acheived if all the intensity values had the same number of pixels -- a flat distribution. This assumes, by the way, that all intensity values are of equal importance -- that light stuff is just as important as dark stuff.
So, to get that, you basically squeegee the values with low numbers of pixels together and separate the ones with large numbers of pixels. If you take the histogram-stretched distribution above and do a histogram equalization on it, you might get something like this (this is done by eye, so *really* doing it would likely be a little different)
Many | | | # of pixels | * | * * * | B * * * * * * | ****** B * * * * * * Few |********** B * * * * * * * ------------------------------------ Dark Light
Brightness
Note that the column with the "B"s is now moved to the left because of the greater space given to the columns with the most pixels. Thus, the ordering of the pixel intensity values is retained, but the relative positions are not.
Finally, there is a method called "local" or "adaptive" histogram equalization in which equalization is not done for the entire image, but only within small local regions. Some algorithms divide the image up into lots of little subimages, and some just look in the region around each pixel. In either case, an equalization is done using only a part of the image. Thus, a dark area in one part of the image will not "use up" all the dark pixels in a bright area. This is useful in the processing of medical images such as CT images. In these cases, the global ordering of the intensity values is lost, but is retain locally.
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Hi, I was wondering if someone could help me with small definitions of what Scanning Transmission X-ray Microscopy and X-ray Imaging Microscopy are. Thanks. Scott
You should microfuge your antibody solutions (especially if they have been frozen or stored for a while). This should get rid of the floaters that fluoresce and clutter up your field.
On Fri, 14 Nov 1997, Arthur Schuessler wrote:
} Date: Fri, 14 Nov 1997 18:18:08 +0100 } From: Arthur Schuessler {schueslr-at-sun0.urz.uni-heidelberg.de} } To: Microscopy-at-sparc5.microscopy.com } Subject: immunofluorescence - UNICRYL-problems } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists, } } since we have a serious problem with immunofluorescence, I hope somebody } can help us. } We used 0.5 and 1 micron sections of Unicryl embedded plant tissues for } immunofluorescence. Sections were cut with an diamond knife / ultracut on } water. } We did labelings with second Antibodies (anti-rabbit) conjugated to FITC or } Cy3. Once we had nice pictures, but 10 times we had } --} } } } horrible spots ("stars") as background all over the regions the } incubation drop was located, and very weak labeling. Also there are } fluorescing droplets when using Cy3, I wonder if it is not ok. However, the } FITC conjugated also shows the "stars". It really looks very bad - all is } full of "dirt". } } What could be the reason? In controls without first antibody the sections } are always clean. We use TBS with 0.1 or 1% Casein, polylysin coated or } uncoated coverslips, heat dried or not heated etc. With no first antibody } always no stars. Is the antigen floating around? } } Thanks a lot if there are any ideas ... } Arthur } Dr. Arthur Schuessler } University of Heidelberg } Zellenlehre } D-69120 Heidelberg } Germany } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
In the process of a move to our new facilities, our phones (which were due to ring in both offices) have developed a mind of their own. We apologize for any inconvenience that this might have caused. If you need to get in touch with Barbara Foster or Ken Piel, please send us an email message with your phone number and we will respond promptly. for Barbara Foster: mme-at-map.com for Ken Piel: kenpiel-at-map.com Emergency phone: (413)736-4828
Many thanks for your patience.
Barbara Foster MME Moving from 53 Eton Street, Springfield, MA 01108 to 125 Paridon Street, Springfield, MA 01118
Phone: still (413)746-6931 Fax: still (413)746-9311 (we think!)
Hi everyone I am looking the buy a variable pressure SEM in the near future. My question to you all is this - A few companies use the chilled differential pumps and state that this type of pump is best when the SEM is in the Variable Pressure mode while others use the Turbomolecular pumps which require only air as it's cooling mechanism. I am asking What is the difference between the pumps, which pump should be use when in VP mode and why?
The kind of problem Jill Craig describes can appear when Windows (3.1.x) runs into the limit of system resources. This can, for example, be caused=
by some application programs that don't free system resources after being=
terminated, or, as Warren Straszheim points out, by faulty hardware (memo= ry etc.). To check out the systems available resources, close all applications and from the Program Manager select the Help menu and About Program Manager. The window that appears will show the currently available system resource= s. If this value is below 30%, even with no application open, some program grabbed a lot of memory and didn't free it after closing. If you restart=
Windows, resources will be available again and everything works normally,=
until you use again the program in question and afterwards any other application could generate the General Protection Error.
There is one easy way to determine if it is hardware-memory related (chip=
failure): when Windows reports the General Protection Error, one can quit=
Windows and immediately afterwards run Defrag (form root directory). Defr= ag will test the system memory on start-up and generate a warning when it finds faulty memory. If it finds no problems in memory, just exit Defrag or, even better, run it in =
full-optimization mode (can only do good to your system). If it does find a problem, it would indeed be best to buy new memory.
However, there is another, much simpler possibility: a bad contact on the=
memory modules themselves. Now, before you go out and buy new SIMM's, you=
may try this: with the computer disconnected form the mains, take the SIMM's out of the= ir socket (avoid any source =
of static electricity when attempting this, wear cotton clothes etc.) and=
clean the module's contacts with the eraser of a pencil (a few passes wil= l do). Wipe off the eraser residues with a tissue and insert the SIMMs agai= n into their sockets, making sure that they are properly seated. This may well solve the problem.
Sorry for the large explanation. I hope it's useful.
*Received: from SpoolDir by IM-PW (Mercury 1.13); Tue, 13 Jan 98 22:26:13 -180 *0 *Return-path: {Microscopy-request-at-sparc5.microscopy.com} *Received: from Sparc5.Microscopy.Com by mailer.inmat.pw.edu.pl (Mercury 1.13) *; * Tue, 13 Jan 98 22:26:10 -1800 *Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id * NAA08918 for dist-Microscopy; Tue, 13 Jan 1998 13:35:54 -0600 *Received: from mail.ucsf.edu (mail.ucsf.EDU [128.218.95.23]) by Sparc5.Micros *copy.Com (8.6.11/8.6.11) with ESMTP id NAA08915 for {microscopy-at-Sparc5.micros *copy.com} ; Tue, 13 Jan 1998 13:35:52 -0600 *Received: from JanLab (u226-mac-01.ucsf.EDU [128.218.106.131]) * by mail.ucsf.edu (8.8.7/CDR8.8.7) with SMTP id LAA25512 * for {microscopy-at-Sparc5.microscopy.com} ; Tue, 13 Jan 1998 11:35:45 -0800 ( *PST) *Message-Id: {199801131935.LAA25512-at-mail.ucsf.edu} *X-Sender: mishot-at-itsa.ucsf.edu *X-Mailer: Windows Eudora Light Version 1.5.2 *Mime-Version: 1.0 *Content-Type: text/plain; charset="us-ascii" *Date: Tue, 13 Jan 1998 11:18:20 -0800 *To: {microscopy-at-Sparc5.Microscopy.Com} *From: Larry Ackerman {mishot-at-itsa.ucsf.edu} *Subject: Ergonomic Microscopy
Well, this is a good point. It seems to me that the ergonomy of the products depends on the manufacturer and the products itself. Working over twenty years with two TEMs, one made by Philips and other one by Jeol, I may say that the Philips manufactured microscope is much more ergonomic. One should take into account that these two are very old and now might be different. I don't expect a huge change, for the "tradition" and, so called, good name play an important role in such case. On the other hand, some time ago, I've been working with Philips CM30 and Jeol 3010 and my feeling stays the same.
I should maybe add that I don't have any financial interest in posting this msg, just personal feeling.
Witold Zielinski Warsaw University of Technology Narbutta 85, 02-524 Warsaw POLAND
I am interesting in the negative staining technique on whole-mount preparation and I have some questions:
1) I would like to know the mechanism by which the contrasters (PTA, Uranyl acetate etc. ) interact with the biomembranes 2) What kind of artifacts could the contraster induce and how to avoid them ? 3) and the same question about the putative artifacts induced by the air drying
Dr. Roberto Weigert Consorzio Mario Negri Sud Department of Cell Biology and Oncology Molecular Neurobiology Laboratory Via Nazionale S.M. Imbaro, 66030 Chieti Italy Phone: 0039-872-570-354 Fax: 0039-872-578-240
At 02:08 AM 1/14/98 -0500, Hermann Reese wrote: } There is one easy way to determine if it is hardware-memory related (chip } failure): when Windows reports the General Protection Error, one can quit } Windows and immediately afterwards run Defrag (form root directory). Defrag } will test the system memory on start-up and generate a warning when it } finds faulty memory. If it finds no problems in memory, just exit Defrag } or, even better, run it in } full-optimization mode (can only do good to your system). } If it does find a problem, it would indeed be best to buy new memory.
I will have to try this test at my next opportunity, although I am somewhat doubtful that it will catch the error. I mentioned that I had used a variety of utilites to check memory besides the initial memory test at boot up and the himem memory test. I used a memory check within WinProbe and a couple of DOS utility sets. They all reported the memory to be fine. However, certain Windows applications (like MS Word) exercised the memory in a more stringent manner and caused failures. Perhaps the newer crop of utilities does exercise memory in the manner that 32-bit apps do and would catch such problems.
And like Hermann said, running DEFRAG is a good thing anyway. Probably one of the overlooked reasons for system slowdown. And his comments about resource leaks is a good one. I know older versions of Microsoft Excel and Trumpet Winsock for Win 3.x had such problems. I can pass along a shareware app that can help you track the resources by type if it would be helpful to anyone. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
Have you tried sputter coating a carbon filmed TEM grid? A light coating should be thin enough for TEM and the grid should be easy to prepare if you have a sputter coater in the lab.
Joe Neilly Microscopy and Microanalysis D-45M Abbott Laboratories AP31 Abbott Park, IL 60064
In the DATABAR dialog box (under the In/Out pull down menu) there is a check box that allows you to remove the black background. Try removing the background and saving a file and see if the the problem is eliminated. This would give you a temporary fix until the problem is solved. It may also indicate that the problem is in the microscope control software.
Dear Microscopists I have been asked to examine a sample of Co/Cu sputter deposited onto a Ga/As wafer substrate. The film is apparantly 50-100nm thick so should be electron transparant, but what is the best method of removing the film from the substrate and getting it onto a grid? I am a bit of a novice with this type of sample, having trained as an em operator in biological sciences and recently transferred to a materials science dept, and thus far have only dealt with alloys which can be electropolished. I thought it might be possible to remove the film by plunging into LN2 but I'm not sure if this will work. All advice will be much welcomed.
Cheers Nikki
**************************** Nikki Bock EM technician Dept. of Materials Engineering & Materials Design University of Nottingham (0115) 9513759 emznjb-at-emn1.nott.ac.uk
There is some sort of minor software problem with our Edax computer. A few times a day, during heavy use, the computer will declare a general protection error. The only way out of this error is to close all programs and exit windows and then restart. After restart nothing appears to be strange, until several hours later another general protection error occurs. I have been looking for a pattern, but have been unsuccessful
I experience "general protection error" after I have an upgrade of my computer. The pc-support guy tried all the tricks he had and could not solve the problem. I finally seek second opinion and the problem was solved.
The problem was that the cd drive was not set up as a slave drive. I hope this helps.
Ann Fook Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Agriculture Agri-Food Canada, Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
I am looking for suppliers of low-temperature modules for doing CRYO-SEM on a JEOL or Hitachi field emission SEM. I am very interested in getting feedback (positive or negative) on cost, reliability and ease of use from those who have experience with such equipment. Manufacturers are welcome to contact me offline. Thank you very much in advance for your help.
Regards,
Kirk J. Czymmek Biological Electron Microscopy Facility University of Delaware email: kirk-at-udel.edu phone: (302) 831-1158 FAX: (302) 831-2281
I wanted to thank everyone with their suggestions for placing sections on formvar slotted grids. We were trying to mount rather large sectins onto the film. The suggestions and a little practice produced excellent results. Hank Adams EML New Mexico State University
Dear Microscopists, Hi - I am new to this area but hopefully have a simple question for you. How do you separate Epon resin from glass? Thank you, James Mulroy
The Microscopy Society of America (MSA) and the MSA Technologists' Forum are the sponsors of the Professional Technical Staff Awards (PTSA) to provide assistance on a competitive basis to full-time professional staff who submit papers for presentation at Microscopy and Microanalysis '98. The following information also appears in the Registration Bulletin and Call for Papers.
Awardees will be selected based on the quality of a first-authored paper submitted for presentation at Microscopy and Microanalysis '98. It is the intent of this award to stimulate attendance for professional technical staff who ordinarily might not participate in the meeting, and to encourage employers to support their staff in professional activities. The awards consist of free full registration for the meeting, a copy of the Proceedings and the Sunday evening social event. MSA will reimburse awardees up to $600 for travel, lodging and other expenses. Applicants must be full paid-up members of MSA at the time of application. Abstracts will be judged by the MSA Technologists' Forum. The applicant must be the first author of the submitted paper. There will be four awards, two in the biological sciences and two in the physical sciences. Successful applicants must present their papers personally at Microscopy and Microanalysis '98 in order to receive the award. They are expected to attend and participate in the entire meeting. Former winners will not be eligible for another award. Applications shall consist of: (1) A copy of the Abstract and Data Form to be sent to the Technologists' Forum committee by FEBRUARY 1, 1998 for judging. Judgment will be made and awardees notified by February 15. Those not receiving awards will be notified in time to retract the original Abstract and Data Form, if desired; [NOTE: An original Abstract and Data Form must be submitted to The Meeting Managers by FEBRUARY 1, 1998 as well.] (2) A supporting letter from the applicant's employer, manager or supervisor, attesting to the applicant's status as a full-time, professional staff member. Send a copy of the Abstract, a copy of the completed Data Form, along with the supporting letter from your employer, manager or supervisor, to arrive by FEBRUARY 1, 1998 to the Chair of the Technologists' Forum: Ms. Beverly Maleeff, SmithKline Beecham Pharmaceuticals, Toxicology-US, UE0462, 709 Swedeland Road, King of Prussia, PA 19406; Phone: 610/270-7987, Fax: 610/270-7202, E-mail: Beverly_E_Maleeff-at-sbphrd.com. The award will be presented at the Presidential event.
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Reply to: RE} Separating Epon resin from glass
Dear James, You may want to try running some liquid nitrogen over the glass to cause = it to break away from the epon. This works for glass coverslips on flat = embedded epon samples. And remember, wear safety glasses and appropriate = gloves for cold temps. while doing this. Good Luck. Linda Chicoine Yale Univ. Center for Cell Imaging New Haven, CT USA
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Dear Microscopists, Hi - I am new to this area but hopefully have a simple question for you. How do you separate Epon resin from glass? Thank you, James Mulroy
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Without a doubt the best and most reliable cold stage for an SEM including a FEG SEM is the CM1500 made by Oxford Instrument. All other manufacturers pale into insignificance. I speak from experience and you are welocme to come to Cambridge and have a go with our system on a Phlips FEG-SEM
Best wishes
Patrick Echlin Director, Multi-Imaging Centre School of Biological Sciences University of Cambridge UK
On Wed, 14 Jan 1998, Kirk J C Czymmek wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear Cryo-microscopists, } } I am looking for suppliers of low-temperature modules for doing CRYO-SEM } on a JEOL or Hitachi field emission SEM. I am very interested in getting } feedback (positive or negative) on cost, reliability and ease of use from } those who have experience with such equipment. Manufacturers are welcome } to contact me offline. Thank you very much in advance for your help. } } Regards, } } Kirk J. Czymmek } Biological Electron Microscopy Facility } University of Delaware } email: kirk-at-udel.edu } phone: (302) 831-1158 } FAX: (302) 831-2281 } } }
Roberto, The interaction of the negative stains which you have in question is as follows: 1. Uranyl Acetate acts as a mild fixative stabilizing the lipids within the membrane, It will bind to carboxyl and phosphate groups to some extent. The specific protein interactions are detailed in several handbooks covering staining sectioned materials. Artifacts are usually a needle shaped precipitate= indicating the stain is too old. We make ours when we are going to use it and never save it more than half a day. 2. PTA has little defined interaction with the membrane. The stain occurs much as a snowfall over the features of the sample. Artifacts include clumps of stain which can be removed with a light wash with distilled water. Our experience has taught us to pH the stain with ammonium hydroxide rather than the customary potassium. A 2% solution will last for as long as 12 months without any problem. The disruptive nature of the NHPTA is far less significant than the KPTA and the granular dimensions are finer give higher resolution. We also see better penetration into the fine details of surfaces similiar to what one sees with ammonium molybdate.
To address the air drying one would need to know the composition of the specific sample. Lability to air drying has been documented in handbooks detailing working with whole cells for SEM. Viruses and bacteria are not as subject to adverse effects from air drying. Some damage can be reduced with a light fixation of the sample after it has been attached to the grids and before drying begins.
Hi James, I had a lot of trouble removing glass coverslips from epon resin. I tried immersing the epon-glass in liquid nitrogen without any luck. Finally I made sure that I "undercooked" the epon. Then, I was able to remove the glass from the "soft" epon after immersing the combo in liquid nitrogen. In fact I nearly injured myself- the glass really flew off at me. From now on I will wear a face shield. After I removed the glass, I had to put the epon back in the oven to finish the polymerization process. You also can heat up the epon-glass combo before dumping the combo into liquid nitrogen.
Be careful, Sally
On Wed, 14 Jan 1998, James Shannon Mulroy wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists, } Hi - I am new to this area but hopefully have a simple question for you. } How do you separate Epon resin from glass? } Thank you, } James Mulroy } } }
Linda's suggestion to use liquid nitrogen is a good one. I've had very good success with this method. Separation is alot easier if the glass is pre-coated with either a siliconizing solution or spray or better yet a very thin coating of evaporated carbon. Also, try to keep the epon from spilling over the edge of your glass surface and wicking underneath. This makes separation more difficult. Good luck. Vickie Frohlich ******************************************************************************** Victoria Centonze Frohlich Deputy Director, IMR University of Wisconsin, Madison P(608) 263-6288 F(608) 265-4076 ********************************************************************************
} Reply to: RE} Separating Epon resin from glass } } Dear James, } You may want to try running some liquid nitrogen over the glass to cause } it to break away from the epon. This works for glass coverslips on flat } embedded epon samples. And remember, wear safety glasses and appropriate } gloves for cold temps. while doing this. Good Luck. } Linda Chicoine } Yale Univ. } Center for Cell Imaging } New Haven, CT USA } ----------------------------------------------. } } Dear Microscopists, } Hi - I am new to this area but hopefully have a simple question for you. } How do you separate Epon resin from glass? } Thank you, } James Mulroy } } }
******************************************************************************** Victoria Centonze Frohlich Deputy Director, IMR University of Wisconsin, Madison P(608) 263-6288 F(608) 265-4076 ********************************************************************************
Dear Microscopist; R.J. Lee Group, Inc is now offering a course on computer controlled scanning electron microscopy. This is an intensive hands-on course (no more than three student per microscope) to introduce you to or improve your skills on the Personal SEM. Much of the instruction is performed on sample material that the students are requestedto provide in advance of the course. Topics covered include: image acquisition, variable-pressure options, file saving and printing options, image functions, optical pre-review, semi-quantitative analysis, computer-controlled analysis (Automated Feature Analysis), routine maintenance, and sample preparation. With the small class size, the course can be matched to meet the participants' specific analytical needs. Over 100 pages of lecture notes (including laboratory exercises), a computer program illustrating various Graphical User Interface (GUI) functions, and a book (BASIC SEM - An Introduction to the PERSONAL SEM by Fred Schamber0 are provided. R.J. Lee Group will be conducting the course quarterly (Feb 16-20, 1998; May 18-22, 1998; Aug 17-21, 1998) at the RJ Lee Group facility in Monroeville, Pa, and additional courses can be scheduled if there is a need. To enroll please contact Barbara Smith at RJ Lee Group (412-325-1776, bsmith-at-rjlg.com) or Doris Allison at RJ Lee Instruments (412-744-0100, allison-at-sgi.net) or reply to this email message and I will pass the information on to them. Also, if you have any course-specific questions feel free to contact Dr. Steven Kennedy at RJ Lee Group (412-325-1776, skennedy-at-rjlg.com).
-- Michael Mizell 412-744-0100 x224 RJ Lee Instruments, Ltd. 412-744-0506 Fax 515 Pleasant Valley Rd. mizell-at-sgi.net Trafford, PA 15085
Dunk it into liquid nitrogen for a half a minute, let it warm and it will just peel off. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} } Dear Microscopists, } Hi - I am new to this area but hopefully have a simple question for you. } How do you separate Epon resin from glass? } Thank you, } James Mulroy } }
Echoing the bad RAM possibility, a friend who works in the computer business is convinced that 90% of computer problems can be blamed on bad or intermittently flaky RAM. However, I have also read recently that the Microsoft IE4.0 cache can (with the default setup) consume vast quantities of hard drive real estate. As the hard drive runs out of room, the Windows system cache gets squeezed. Those problems described in the original post can also be caused when the Windows cache is insufficient. This problem might be eliminated by simply clearing the IE4.0 cache or by changing the settings to control the cache. Kevin Brent Smith University of Louisville Biology Dept.
Dear Nicola, When I am asked to analyse something like this, I always ask the researcher to stick a carbon-film-coated TEM grid onto his Ga/As substrate, so it intercepts the sputtered film. I have had good success using this method to study sputtered films. In your case, where you want to remove a film already in place, the classic Materials Microscopy method is the etch the substrate with a suitable chemical etchant, which will release the film. I am not sure if the Co/Cu film will hold together on its own, the usual is to sputter onto a carbon film, which helps hold it together. But you can try, amd pick up any film that floats up off the substrate. You wrote: } Dear Microscopists } I have been asked to examine a sample of Co/Cu sputter deposited onto } a Ga/As wafer substrate. The film is apparantly 50-100nm thick so } should be electron transparant, but what is the best method of } removing the film from the substrate and getting it onto a grid? } I am a bit of a novice with this type of sample, having trained as an } em operator in biological sciences and recently transferred to a } materials science dept, and thus far have only dealt with alloys } which can be electropolished. } I thought it might be possible to remove the film by plunging into LN2 } but I'm not sure if this will work. } All advice will be much welcomed. } } Cheers } Nikki Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
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The Oxford system is of excellent quality, but you should request for information on the new CRESSINGTON system. I consider it as an instrument of a new technical generation. In particular when you are interested in real high resolution work it is a must to inform yourself about the Cressington.
Contact Dr. P. Walley, Cressington Scientific Instruments Ltd, Tel. 1923 220499 in the UK, or A. Berginc, Tel (412) 772-0220 in the USA
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Without a doubt the best and most reliable cold stage for an SEM including a FEG SEM is the CM1500 made by Oxford Instrument. All other manufacturers pale into insignificance. I speak from experience and you are welocme to come to Cambridge and have a go with our system on a Phlips FEG-SEM
Best wishes
Patrick Echlin Director, Multi-Imaging Centre School of Biological Sciences University of Cambridge UK
On Wed, 14 Jan 1998, Kirk J C Czymmek wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear Cryo-microscopists, } } I am looking for suppliers of low-temperature modules for doing CRYO-SEM } on a JEOL or Hitachi field emission SEM. I am very interested in getting } feedback (positive or negative) on cost, reliability and ease of use from } those who have experience with such equipment. Manufacturers are welcome } to contact me offline. Thank you very much in advance for your help. } } Regards, } } Kirk J. Czymmek } Biological Electron Microscopy Facility } University of Delaware } email: kirk-at-udel.edu } phone: (302) 831-1158 } FAX: (302) 831-2281 } } }
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On Wed, 14 Jan 1998, NICOLA BOCK wrote: Hi Nikki,
I assume that you don't have a carbon underlayer on your substrate, in which case you are unable to float the film off. Try sticking a small piece of sellotape firmly onto the film and quickly ripping it off. This should remove some film and the sellotape can be dissolved on chloroform and the film pieces picked up from the solvent onto a grid.
Good luck. Ron
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists } I have been asked to examine a sample of Co/Cu sputter deposited onto } a Ga/As wafer substrate. The film is apparantly 50-100nm thick so } should be electron transparant, but what is the best method of } removing the film from the substrate and getting it onto a grid? } I am a bit of a novice with this type of sample, having trained as an } em operator in biological sciences and recently transferred to a } materials science dept, and thus far have only dealt with alloys } which can be electropolished. } I thought it might be possible to remove the film by plunging into LN2 } but I'm not sure if this will work. } All advice will be much welcomed. } } Cheers } Nikki } } **************************** } Nikki Bock } EM technician } Dept. of Materials Engineering & Materials Design } University of Nottingham } (0115) 9513759 } emznjb-at-emn1.nott.ac.uk }
========================================================================== = Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ========================================================================== ==
Where the microscope is in the Variable pressure mode, the pressure inside the specimen chamber is comprised between 1 to 270 Pa. The low vacuum pumping line is only composed of rotary pump. I wrote a paper in microscopy and analysis intitled "priciples and applications of the vpsem". reference C mathieu european microscopy and analysis, september 1996, 43,13 If you want I can send you this publication.=20
Yours sincerely,
Dr C mathieu Universit=E9 d'artois Facult=E9 jean perrin SP 18 62307 lens cedex France phone 33 3 21791710 fax 33 3 21791755 email mathieu-at-univ-artois.fr
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Sorry to perhaps raise again an old chestnut, but ....
* You may want to try running some liquid nitrogen over the glass to cause * it to break away from the epon. This works for glass coverslips on flat * embedded epon samples. And remember, wear safety glasses and appropriate * gloves for cold temps. while doing this. Good Luck.
Gloves with liquid nitrogen are NOT a good idea. A BRIEF contact with LN2 on skin of hands does little harm, but if it freezes the surface of the glove this may stick to you and give frostbite, and even worse if it gets down inside the glove.
I learned this from a lecture on "The Physics of Ice Cream" by Peter J.Barham of Bristol University. Rapid freezing works !!!
I have now completed the microscopy workstation assessment form for our Intranet system. It was called for as a preventative against the development of postural problems. Once you delve into this, it appears that these are not uncommon in our line of work. They are best avoided. Basically, it is a safety check-list where a "No" response can be detected automatically to flag a problem that someone needs to address.
The form for use is a four-column table with "Yes" coming before the "If, no" and "No" columns. The idea is to prompt people to sort out their problems themselves where possible, before answering "No". The version below is simply more readable in e-mail. Maybe you can copy it and format it further in a WP package.
Alternatively: it is available as a MS Word 6 file and there is also a wonderful new drawing showing how to sit at a microscope (by me!). I hope it is correct. This is available in Windows Draw 3 .drw and TIF format, from {k.ryan-at-pml.ac.uk} .
I will send these files automatically to those acknowledged in the Acknowledgemnts section below. Many thanks to those who responded via the "Microscopy", "Histonet", Safety, and "Sorehand" Lists.
My background is 29 years in microscopy (mostly EM) and 5 years as a safety advisor (part-time). I knew something "official" about setting up pc workstations, but reviewing microscopy set-ups was interesting.
"Thank you" to those who responded, I picked up a new point or two! (old dogs and new tricks?) ______________________________________________________ e-mail version Workstation assessment for Safe Use of Microscopes
Introduction. Looking through a microscope for extended periods is not what we were designed for. It requires holding our bodies in an unnaturally rigid position. An ergonomically designed microscope would have eyepieces at about 90* from vertical. It is important to adopt a correct, ergonomic working posture. This means fitting the workstation to the worker, not vice versa. It is also important to take regular breaks.
Ideally, the microscope should be on a bench which is adjustable for height: first, the seating position is adjusted (steps 2-8 below) followed by the bench height and subsequent steps (11-22 below). The following is based on a fixed work bench. See Fig. 1.
Before entering *No' to the following questions, attempt to rectify the problem.
1. Have you been show how to use the microscope, including how to align the optical path to optimise performance? If no, seek this training before continuing. Otherwise, poor results and eye strain may ensue.
2. Are you sitting back in the chair, rather than perching on it? If no, sit back into the chair.
3. Is the height of the chair adjusted so that your feet are resting comfortably, flat on the floor? If no, adjust the chair height appropriately.
4. Is there an even pressure along the backs of the thighs? If no, check if the seat platform can be tilted appropriately for comfort. If not, consider readjusting the height slightly.
5. Does the chair support your back in an upright position? Ideally, it should support to beyond the level of the shoulder blades. If no, adjust chair back appropriately.
6. Does the chair back give support in the lumbar region? If no, check the chair's back adjustment again, possibly adjusting the tilt control. Or, obtain a separate lumbar cushion.
7. Are you now sitting with your back upright? Ask a colleague to comment. If no, repeat steps 2-6 above.
8. Are the microscope eye-pieces in line with, or extending over, the front edge of the bench? If no, move the microscope towards you appropriately (caution - you may need skilled assistance)
9. Is the vertical position of the eye-pieces a little high for comfort, so that your head is upright? Initially, this will feel unnatural. If no, raise the microscope vertically to a suitable height at which you are are forced to sit upright. This can be done with layers of plywood etc. If you are using the microscope long-term, get the workshop to make a suitable stand.
10. Can you see at all into the eye-pieces of the microscope? If no, raise the chair height appropriately and obtain a suitable footrest.
11. Are you gazing slightly downwards into the eye-pieces, as opposed to tilting your head and *looking straight-ahead' into them? If no, you are not sitting upright enough. The back should be *vertical' and the neck and head upright. Holding the head tilted for long periods will induce neck and shoulder-ache. Repeat steps 2-10.
12. Is the leg-well clear of clutter so that your legs and feet are not impeded when sitting at the bench? If no, clear the clutter.
13. Are your thighs clear of the under-surface of the bench? If no, the bench is unsuitable for microscopy work. Have it modified or seek another site for the microscope.
14. When operating the focus and stage controls, are your forearms resting on something, either the bench or microscope arm rests? If no, obtain arm rests, perhaps sloping. Holding the arms off the bench for long periods will induce static loading problems. The most comfortable position for the hands is as for when shaking hands. A commercial source for arm rests is: http://members. aol.com/rdergo2/msm.htm.
15. Are the eyepieces set correctly for your inter-pupillary distance? If no, set this distance properly - the oculars should move towards or away from each other. This reduces eye-strain.
16. Are the eyepieces parfocal? If no, adjust them individually so that the image is sharp in each. This reduces eye-strain.
17. Are the eyepieces clean, for both optical and hygeine reasons? If no, ensure that they are cleaned. Be aware that communicable eye diseases such as conjunctivitis can be transmitted by contact.
18. Before you begin microscope work for the first time, are you free from pre-existing visual problems? If no, you should see an optician in case you have astigmatism, fusion insufficiency (poor eye co-ordination) or simple long/short sight. Microscopy work may make these problems more obvious.
19. Are your surroundings free from glare and reflections? If no, try to remove light sources from the visual field by re-positioning the workstation, removing highly reflective surfaces, using blinds, curtains or other screens.
20. Is the image in the microscope free from glare and reflection? If no, adjust the internal lighting so that there is not an uncomfortably high level of light or contrast. This may be done by regulating the transformer or by using appropriate filters.
21. Are you satisfied with other environmental factors, such as temperature, humidity, draughts, ventilation, ambient lighting? If no, try to sort problems locally yourself or discuss them with line management. Beyond that, see your union safety representative or local safety advisor. Humidity (and dry eyes) are aided by watering plants, where appropriate.
22. Will you be focusing your eyes to distant vision periodically, e.g. through a window? If no, you should take regular breaks from the work and look at something distant. This is prevent headaches/eye strain
23. Are you/will you be taking regular breaks from the microscope, e.g. two or three minutes every half-hour, or rotating jobs? If no, this needs urgent consideration. Breaks are necessary to prevent RSI (Repetitive Strain Injury), WRULD (Work-Related Upper Limb Disorders) or CTD (Cumulative Trauma Diseases). Discuss it with your manager, union safety representative or local safety advisor. Computer users are recommended to take five minutes every hour, microscopy work is probably more posturally demanding.
24. When taking breaks from the microscope, will you be doing stretching exercises? If no, refer to the article *Applying ergonomics to improve microscopy work* by Helen Haines and Lyn McAtamney in Microscopy and Analysis, Issue 36 (July 1993), 15-17 (available from your local safety advisor). You should do these exercises to relieve the static loading fatigue/stress on the body. This applies equally to computer users.
Acknowledgements: this assessment document was compiled with reference to the UK Health & Safety (Display Screen Equipment) Regulations 1992 and the article *Applying ergonomics to improve microscopy work* by Helen Haines and Lyn McAtamney (University of Nottingham, UK). Members of the *Safety*, *Sorehand*, *Microscopy* and *Histonet* Internet Lists also made helpful comments, particularly Bob Chiovetti (E. Licht Co., USA), Barbara Foster (Pres., Microscopy/ Microscopy Education, Springfield, MA, USA), Dan Kallin (Bose Corporation, Framingham, MA, USA), Bob Morency (R&D Ergonomics Inc, Freeport, ME, USA), Philip Oshel (Champaign, IL, USA), Sue Reilly (James Cook Univ. NQ, Australia), John Shane (McCrone Research Institute, Chicago, USA), and especially Stephen Shaffer (MicroDataware, Berkeley, CA, USA).
Dear James, the problem you describe seems to have a solution named = "pop-off"-technique. Depending on glass quality of your object slide (not all brands seem to = be the same quality and therefore display variable physical properties) you can separate postpolymerized (e.g. EPON)-specimen blocks of varying area = size from object-slides, including a selected area of your former semithin = section or the whole area (as used in reembedding techniques, which I assume is the purpose you have asked the question).
If your concern is: =20 Re-embedding of (immuno-) histochemical reacted LM-sections from e.g. = thick paraffin section (10-40 =B5m):
after several steps of processing like: incubations, washing, = osmication, dehydration,----intermedium------intermedium/resin-----resin pure: =20
if you place a polymerized resin block over the selected section area ( = e.g. also reembeddings of bigger LM-/Histological sections), be sure that you have flushed/ infiltrated the area repeatedly with fresh resin (complete with = hardener/accelerator) after an intermedium/solvent (e.g. = propyleneoxide-PO, PO:resin, etc...) step to get rid of any trace of solvent. This repeated = exchange of resin should be done by only dropping fresh resin into the middle of = the section area to be re-embedded: the former resin would be transferred to = the outer margins of the area to be re-embedded.
Polymerize the resin block (with a clean! and plane/smooth block-face) = on your section to be re-embedded for correlative LM/TEM at least for 1.5 hours = at only 37 degr. C., then you are up to polymerize at higher temperatures, say 1 = h at 80-90 degr. C.
Let cool down the "combo" of resin block-intercalated section-glass = surface to room temperature.
Prescore by means of a scalpel blade (be careful) the four sides of the = resin block (try to cut through resin remnant "walls" down to the glass slide = surface).
If you were lucky with that without hurting you (depends on the = scalpel-knife- handle and the eye protection you should use), two choices:
first: =20 -(faster) flat container made from styrene foam is filled about 1 cm = high with LN2 (liquid nitrogen) -dip ONLY the object slide in the LN2, sufficiently long (usually until = evaporation and bubbling of N2 Gas has stopped). If the surface of the object slide = gets a little bit of LN2 it wouldn't matter.=20 -put out of the LN2 the "combo" and either let warm up (resin block in = between your fingers !): at a time you will hear the "popping click" or try = then to apply "shearing force" to the block to separate the resin block from = the object slide.
second: -(needs another time for warming up your "combo" in an oven): warm up = the "combo", say up to 80 degr. C.for about 10-15 minutes, then, proceed as above.
NOTE: depending on the quality of glass slides (physical properties) and (individual) technique used the separation process is or may not be not uniform, i.e. there = sometimes is left some glas particles on the block surface after separation or no = separation at all. Elegant separation normally takes a few tests or trials. So = don't use important or valuable tissue/section preparations for your first or = second attempt. If nothing changes and separation is not performed think of a spraying = or a "finely dispersed pasting" of your object slides using separation media = (e.g. Teflone-spray, silicone pasting) before mounting your sections to be = re- embedded. Another *important* point is that if you're dipping the whole "combo" = (resin block- at least at its end- connected to the glass slide) into the LN2 = to cool it down you might have big problems due to *cracking* of the resin block = which might interfere with your considerations on cutting the whole areas you selected to be sectioned (I have done "reembeddings for TEM" of 2 =B5m = to 40=B5m thick immunohistochemically reacted tissue cryosections = =3D=3D=3D"pre-embedding labeling techniques"=3D=3D=3D=3D=3D up to section areas of 5x5 mm, and = it worked most of my attempts, but: it is very, very tricky and good results seem to = depend on the day-time, or at least on how you stood up in the morning (you know: = left foot first!).
The effect of separation ("pop-off-technique") is explained by the = difference of the coefficient of expansion of glass versus resin. Since glass has another coefficient compared to resin formulations, glass contracts on low = temperature and expands on warming up in another fashion like the resin used and = vice verse.
Hope this helps anyway and doesn't come too late If any questions, please contact me by e-mail.
Best regards Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
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Dear Microscopists, Hi - I am new to this area but hopefully have a simple question for = you. How do you separate Epon resin from glass? =20 Thank you, James Mulroy
For those who are DEEPLY involved in SEM there is a course which will interest, inform, and from past experience excite!
The course called "INTENSIVE SEM" takes one week (20 to 24 April), is held at the University of Surrey, England, and comprises of morning lectures and afternoon practicals.
Unlike any other course that we know of only four students may take part, it is a matter of one lecturer and four students for one week investigating almost every avenue of SEM performance,
The course covers starts from a basic level, just to make sure we all really know about the instrument structure, and then moves on through to advanced levels, covering in the practicals - what really makes the instrument tick, how does kV effect resolution and information, how can we improve low kV performance, just what does manipulating the gun geometry do for helping make the impossible possible, how specimen-detector geometry (WD, tilt, detector style) effects performance and information, high resolution SEM, calibration of drift rate, of contamination rate and general performance monitoring, looking at ANY specimen uncoated, and preparation techniques that make life easier.
Not a course for the faint hearted, this is hard work, but an information gathering exercise that SEM devotes will love. People talk of hands on courses this is one when you hardly have time to rest those hands!
Details from protrain-at-compuserve.com Telephone or Fax 44 + (0)1844 353161 More data on http://ourworld.compuserve.com/homepages/protrain
If you have access to dry ice set the coverslip or slide on the dry ice for 2-3 minutes, the resin block will pop off with no damage to specimen. If you avoid excess resin on the glass, this method will work with several resins. If the glass has been warmed on a hot plate before exposure to dry ice, this procedure is extremely effective.
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
we have a Zeiss EM902A. When using Ni-grids I observe that they stick to the specimenholder. Cu-grids do not. Both are formvar-coated. Has someboby regarded the same phenomenon? Does this mean that the specimenholder is magnetic? Or do you have any other ideas.
Thanks in advance
Birgit
Dr. Birgit Neubohn Institute of Plant Genetics and Crop Plant Research (IPK) Corrensstr. 3 D-06466 Gatersleben-Deutschland
Greetings, Anyone out there know of a dealer who handles Aus Jena stock? As you may know, Aus Jena what was the East German branch of Zeiss was called after the second world war and until the recent reunification of the companies (a few years after the country). I have a stand made by Jena that I'd like a part for, but Zeiss is no longer selling them. Thanks in advance for any leads, Tobias Baskin
Help! My company needs to visually assay highly-swelled particles of cross- linked carboxymethylcellulose in various mixes of water and ethylene glycol (up to 80% EG). The swelled particles are large enough to be easily seen with 10 to 100 power magnification - except that they are essentially invisible due to having virtually the same index of refraction as the liquid. I know this is not an unusual problem, but I do not know how to make the particles visible (except by draining off enough liquid that the particles emerge, which is something we can not routinely do). I assume (hope) that there is some stain that will make these particles visible. If not, perhaps there is some polarizing technique that would make them visible. Any suggestions would be appreciated.
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The 1997 MSA Traveling Exhibit is sponsored by the Education Committee of The Microscopy Society of America. It consists of the top ten posters (five Biological Science and five Physical Science) chosen by a committee of judges at the annual meeting of Microscopy & Microanalysis '97 in Cleveland Ohio. This is an excellent opportunity for our Local Affiliate Societies to receive this exhibit and display it at their local meeting for members to see some "state of the art research" that our colleagues are conducting in facilities throughout the world. The only cost to a local society is to ship the exhibit on to the next destination. I am accepting requests and will try to accomodate as many as possible, while allowing time for shipping. Please contact me as soon as you have a meeting date with your request so that I can place your event on the calendar. This years posters are listed on the MSA web page.
I can be reached via telephone, fax, e-mail or postal mail at the following new address:
Pamela F. Lloyd Research Associate Monsanto Co. 800 N. Lindbergh Blvd. U1E St. Louis, MO 63167 Phone: (314)694-6527 Fax: (314)694-8065 e-mail: Pamela.F.Lloyd-at-Monsanto.com
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James- it kinda depends on the resin & the glass, but if it's epon and a slide, like re-embedding, we use a low heat flame (alcohol burner), hold the slide (with pliers) over the flame 5-10 sec. then use another pair of pliers to snap the capsule off from the glass slide. if you situation is something other than this, you may find some solvents to remove resin from glass. -MR
On Wed, 14 Jan 1998, James Shannon Mulroy wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists, } Hi - I am new to this area but hopefully have a simple question for you. } How do you separate Epon resin from glass? } Thank you, } James Mulroy } } }
26th Annual Symposium of the Florida Chapter of the American Vacuum Society and 16th Annual Meeting of the Florida Society for Microscopy
February 23 - 26, 1998 at University of Central Florida Student Union Building Orlando, Florida
Co-Sponsored by
FL AVS, FSM, UCF, Cirent Semiconductor
Technical Sessions (No Registration Fee-Please Pre-register)
The technical program will be held on Monday and Tuesday February 23 and 24. There will be 6 oral sessions and a joint poster session. A keynote address will start the conference on Monday morning. The rest of the morning session will be on Vacuum Technology in parallel with the FSM Biological Division session. The afternoon will have the AVS session on Thin Films followed by a joint poster session. The Tuesday morning program will be a joint session with the FSM Physical Division and the AVS session on Surface Science and Analysis. The technical program will concludes in the afternoon with the AVS session on Electronic Materials.
For further information contact the session chairpersons:
A short course program will be presented from Monday February 23 through Thursday February 26, 1998. The program consists of 1 or 2 day courses taught by leading experts in their fields. The courses Monday and Tuesday will be at the UCF Student Union and the courses Wednesday and Thursday will be at the UCF Holiday Inn. Here is a list of when the courses will be offered along with the instructors:
An Overview of Applied Vacuum Technology, Woody Weed, Monday and Tuesday Sputter Deposition, William Westwood, Monday and Tuesday CVD for Microelectronics, J. William Rogers, Monday Operation and Maintenance of Vacuum Pumping Systems, Paul Holloway, Wednesday and Thursday Plasma Etching and RIE, John Coburn, Wednesday and Thursday Thin Film Vapor Deposition and Patterning Techniques, Robert Waits, Wednesday and Thursday Vacuum Leak Detection, Rudold (Rudy) Schubert, Thursday Introduction to Contamination Control in Semiconductor Manufacturing Equipment, Gordon Johnson, Tuesday Optical Diagnostic Techniques for Plasma Processing, Gary Selwyn, Monday Surface Preparation for Thin Film Deposition, Gary McGuire, Tuesday An Introduction to Surface Analysis Techniques, John Grant, Wednesday
For further information or to register for short courses, contact Margaret Stringer of the American Vacuum Society at 212-248-0326, 212-248-0245 (FAX), or margaret-at-vacuum.org. Course descriptions, additional information, and electronic registration will be available through the AVS home page, http://www.vacuum.org.
Hotel Reservations
A block of rooms have been reserved at the Holiday Inn UCF, 12125 High Tech Ave., Orlando, FL 32817, 407-275-9000. Please contact the hotel directly to reserve your room prior to the CUTOFF DATE of January 30, 1998. Ask for the special rates for the Florida Chapter of the American Vacuum Society. The room rate is $79 per night plus tax. This rate is available for extended stays from Saturday February 21 through Friday February 27, 1998.
Invited Speakers
Keynote (Monday Morning): Dr. John Hitt, President, University of Central Florida
Monday Morning
Vacuum Technology: Dr. H.F. Dylla, Jefferson Lab Dr. R.E. Ellefson, Leybold-Inficon
FSM-Biological: Dr. Ralph Albrecht, U. of Wisconsin Dr. Clinton Dawes, U. of South Florida Dr. Dale Johnson, U. of South Florida Dr. Johannes Rhodin, U. of South Florida
Monday Afternoon
Thin Films: Dr. Patricia Athey, PPG Industries Dr. James Harper, IBM Dr. Gary McGuire, MCNC Dr. Steve Pearton, University of Florida
Tuesday Morning
Surface Science & FSM-Physical: Dr. Ronald Anderson, IBM Dr. Terry Barr, U. of Wisconsin Dr. Vimal Desai, U. of Central Florida Dr. Robert Hull, University of Virginia Dr. Joseph Michael, Sandia National Labs Dr. Robert Wallace, Texas Instruments Dr. Anthony Warwick, U. of California
Tuesday Afternoon Electronic Materials: Dr. Carlye Case, Lucent Technologies Dr. Joe Greene, University of Illinois Dr. Gerald Lucovsky, NC State University Dr. Bill Rogers, University of Washington
Equipment Exhibit (No Admission Fee)
On Monday and Tuesday, there will be over 40 equipment vendors displaying and describing their latest product lines. The current list of vendors who have reserved booths is:
Spectra International Surface Interface Televac The Sales Advantage Group TOSOH SMD UC Components UCF Varian VAT VWR
Demonstrations of the Hitachi Variable Pressure SEM will be held February 23-25, from 8 am to 5 pm. Contact Dr. Lucille Giannuzzi for details.
Meeting Participant Registration Form
Name:
Company:
Address:
Phone:
Fax:
E-mail:
Days Attending: Monday ( ) Tuesday ( ) Symposium Registration - FREE Monday Night Banquet ($20.): Yes ( ) No ( )
There is no charge for the symposium, equipment exhibit, or lunch (Monday and Tuesday). However, pre-registration is required for getting meal counts, making ID badges, and information packets. The symposium banquet will be held Monday evening at UCF; tickets covering the cost of the meal ($20.00) will be sold at the registration desk. Send the completed form to Rich Irwin, Cirent Semiconductor, 9333 S. John Young Parkway, Orlando, FL 32819, 407-345-6999 (FAX), or irwin-at-lucent.com.
Word 'Aus' is German word and mean 'From'. In the Jena is now headquarters of company Carl Zeiss.
Henrik Kaker SEM-EDS Laboratory Slovenia
Tobias Baskin wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings, } Anyone out there know of a dealer who handles Aus Jena stock? As } you may know, Aus Jena what was the East German branch of Zeiss was called } after the second world war and until the recent reunification of the } companies (a few years after the country). I have a stand made by Jena that } I'd like a part for, but Zeiss is no longer selling them. } Thanks in advance for any leads,
-- Henrik Kaker SEM-EDS Laboratory Metal Ravne d.o.o. Koroska c. 14 2390 Ravne Slovenia Tel: +386-602-21-131 Fax: +386-602-20-436 SEM-EDS Lab http://www2.arnes.si/guest/sgszmera1/index.html MVD Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
I'm posting this as a favor to the individual named below. She asked me for info on possible SEM courses and I was not able to help. Excerpts from her notes are below. She has already contacted Lehigh regarding their courses.
Marisa Ahmed wrote:
I work in the lab of an engineering company in Kanata, ON and want information/training on sample preparation and delayering of IC's (for SEM mostly) for failure analysis-type work. If, as Prof Lyman suggested, you could help me locate such a possible course or resource I would appreciate it.
------------- And then a day later after I told her my course was TEM prep... -------------
You are correct, I am not specifically looking for a course for TEM prep (especially since we don't have a TEM here), although I would be interested in knowing what was included in your course and where it is being taught for future reference. I am actually hunting for something to brush up my SEM sample prep (for topo views and delayering of IC's mostly, not so much cross-sectioning). I also want to learn something about the replica-stripping technique that I believe is used for STEM and some SEMwork for radioactive samples.
If you could post my query I would much appreciate it. Respondants can reach me at mahmad-at-semiconductor.com
Thank-you again,
Marisa
((Sounds interesting. Please also copy me on responses. Ron A.))
Isabel Nogueira wrote: ======================================= I need a gold standard to calibrate an EDS analytical system attatched to a TEM.
I've contacted several suppliers but none of them have anything with gold (pure or in a compound).
Does anyone know where or how I can get a thin film standard of pure gold or of a compound including gold ? ======================================== SPI Supplies has up on our website (for address, see below) a number of different gold "standards" and "aids" for calibration purposes, including Prickley Gold on Copper and our Combined Test Specimen of small gold islands on a carbon film. If the data is taken near a grid bar you will be seeing data from both gold and copper. Micrographs accompany each product listing so you can see exactly what you are getting. We also offer thin microtomed sections of foils spanning the atomic number range for checking with other elements (e.g. besides gold).
Disclaimer: SPI Supplies is a manufacturer and distributor of these standards and calibration items as well as one-off custom made products for others.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
James, Although I can't comment on the use of liquid nitrogen to resolve your present problem I can suggest a possible alternative. In our lab we have been embedding various specimens in Araldite epoxy resin. Rather than glass we deposit the epoxy resin onto another such featureless surface, that being a material made by DuPont having the trade name "TEDLAR", which I am told is a titanized mylar. Once the epoxy has cured it is easily removed from this material. I didn't purchase this material myself and the small roll of this film that we have been using seems to have been around for a number of years. I don't know if your basic mylar sheet would do the trick. Perhaps there is someone out there who can provide further information. If not, try contacting DuPont.
Good luck Paul ----------
Dear Microscopists, Hi - I am new to this area but hopefully have a simple question for you. How do you separate Epon resin from glass? Thank you, James Mulroy
We are fully up and running and I believe we have responded to all the people we know were trying to reach us. If you haven't heard from us and thought you had left a message, please try again at the usual places.
Thanks,
JD Arnott Ladd Research 13 Dorset Lane Williston, VT 05495 USA
TEL 1-800-451-3406 (US) TEL 1-802-878-6711 FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net
I find that by heating the glass, the epon block can be gently pried off. Heat quickly on a warm to hot heat plate for 15 s to 3 m. If you pry too hard, you risk breaking the glass, or pulling a bit of the block off. When warm, the glass will usually pop off readily. It is best not to use the super-adhesive glass slides often employed in immunohistochemistry, - if you have an option.
And of course, the converse, i.e., liquid nitrogen, also works well but infrequently there is a risk of cracking the block.
Peter O. Steele, Ph.D. PMIAC Pathology, All Children's Hospital St. Petersburg, Fl.
Dear Birgit, } } we have a Zeiss EM902A. When using Ni-grids I observe that they stick to } the specimenholder. Cu-grids do not. Both are formvar-coated. } Has someboby regarded the same phenomenon? } Does this mean that the specimenholder is magnetic?
That would be my guess. I have often seen "non-magnetic" stain- less steel become magnetic after machining. You may be able to demagnet- ise the holder, so you might try putting a Ni grid in to see it stick, then de-gauss and see if it comes free. Yours, Bill Tivol
Dear Ray, } } Help! My company needs to visually assay highly-swelled particles of cross- } linked carboxymethylcellulose in various mixes of water and ethylene glycol } (up to 80% EG). The swelled particles are large enough to be easily seen with } 10 to 100 power magnification - except that they are essentially invisible due } to having virtually the same index of refraction as the liquid. I know this } is not an unusual problem, but I do not know how to make the particles visible } (except by draining off enough liquid that the particles emerge, which is } something we can not routinely do). I assume (hope) that there is some stain } that will make these particles visible. If not, perhaps there is some } polarizing technique that would make them visible. Any suggestions would be } appreciated. } If you "stain" the water, i.e., disolve something colored which will not enter the CMC particles, like blue dextran, will that allow them to be observed? Enough solute to make a large change in the refractive index of the water could also work; again, you may have to arrange things so that the solute does not enter the CMC. Good luck. Yours, Bill Tivol
I would like to know your recommendations for a TEM and SEM Film Scanner. The film sizes are 3 1/4 x4" and 4x5" black and white. Any suggestions would be most appreciated. The output will be to a personal computer.
Thank you.
Annette Andrews, EM Lab. PAI -NCTR 3900 NCTR Road Jefferson, AR 72079
I'm looking for a reference ( or an explanation) as to why graphite sheets, in partially graphitised carbon black ( or turbostratic carbon,) have a tendency to curve. Is there a fundamental reason for this ? Is a closed loop structure (energetically) more favorable than a flat sheet configuration with open ends ? I would appreciate any references or comments dealing with this.
Dear Dr. Neubohn, isn't it a great pleasure to have sticky grids *on the specimen holders* = of the EM? You won't loose any grid in the EM itself when viewing. Also you have to grip for them individually for removal from the holder = with tweezers.
Why a pleasure ?(IMO=3D in my opinion): due to static load (of my own, = lab-room, etc.) I saw a lot of grids (even Cu-grids) flying around when = trying to get them into the holder, in and out of the gridbox.=20
More of a "sticking" problem for me were the tweezers I used: Nickel-grids are somewhat curious: if you use "isolated tweezers" = (isolated handles, say for electronic purposes) you will see static = loading and therefore sticking of grids to the tip of tweezers. You = won't be able to detach them readily to the holders grid receptaculum. = Sometimes this also happens if you use "normal" (i.e. made from ordinary = steel) tweezers (mine were originally supplied from ZEISS 17 years ago). = Maybe you will transfer static loading via the tweezer tips to your grid = box, made of plastic: have you seen grids, jumping out of the grid = hole?? To overcome that problem I bought (due to a friends suggestion) curved = tweezers made from "titan". You can purchase such tweezers (especially = if you have to work predominantly with Nickel-grids) (e.g. tweezers, = "Curved tips, Biological, Ti") from the major EM-suppliers (in Germany: = see for instance: PLANO-PLANNET, MARBURG FRG; BALTEC maybe....) If you need addresses of supplier(s), please require by e-mail.
Best regards good luck und "gute Nacht"
Dr. Wolfgang MUSS=20 ("driving license" for a ZEISS (T)EM 109 with "top entry")=20 Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
DISCLAIMER
Disclaimer: The views expressed in this E-mail message do not necessarily represent = the official views of the Dept. Pathology LKA, from which this message = was conveyed. No commercial interest in products/product lines, company/-ies, if such = names are mentioned or such are refered to.
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Hello,
we have a Zeiss EM902A. When using Ni-grids I observe that they stick to the specimenholder. Cu-grids do not. Both are formvar-coated. Has someboby regarded the same phenomenon? Does this mean that the specimenholder is magnetic? Or do you have any other ideas.
Thanks in advance
Birgit
Dr. Birgit Neubohn Institute of Plant Genetics and Crop Plant Research (IPK) Corrensstr. 3 D-06466 Gatersleben-Deutschland
{html} Hi Annette: {br} {br} I use here in the lab "Leafscan 45" which is relatively good. It works {br} with Adobe Photoshop. {br} {br} Regards, {br} {br} -Osman Gurdal {br} {br} -- {br} University of Illinois at Urbana-Champaign {br} 1101 W. Springfield Ave., ESB #1-133 {br} Urbana, IL 61801 U.S.A. {br} Tel: (217) 333-7080 (o) {br} Tel: (217) 355-2933 (h) {br} Fax: (217) 244-1631 {br} E-mail: o-gurdal-at-uiuc.edu {br} URL : {a href=3D"http://www.students.uiuc.edu/~o-gurdal" eudora=3D"autourl"} {font= color=3D"#0000FF"} {u} http://www.students.uiuc.edu/~o-gurdal {br} {br} {/a} {/font} {/u} {font color=3D"#000000"} > {br} >Hello Fellow Microscopists, {br} > {br} >I would like to know your recommendations for a TEM and SEM Film {br} >Scanner.=A0 The film sizes are 3 1/4 x4" and 4x5" black and white.=A0 Any {br} >suggestions would be most appreciated.=A0 The output will be to a=20 {br} >personal computer. {br} > {br} >Thank you. {br} > {br} > {br} >Annette Andrews, EM Lab. {br} >PAI -NCTR {br} >3900 NCTR=A0 Road {br} >Jefferson, AR 72079 {br} > {/font} {BR} {/html}
Hi, all. As you know, graphite is a good sample for AFM and STM while testing their resolution. Recently we want to buy some of graphite samples. But after I contacted with a few graphite companies, none of them would like to sell me a little amount of single crystal graphite sample. (or, they dont have single crystal graphite at all).
Does anyone know where can I buy a little amount of this stuff? Thanks so much and my best regards!
We are using Polaroid SprintScan 45 with very good results. The only drawback in our case, although I read in this list that some people solved this with Polaroid, is that the negative holders supplied with the equipment are not exactly the size of our negatives. We us a larger holder that helds the negatives only between 2 sides of the holder and the negatives almost do not bend. We decided to buy this scanner because, after testing some high resolution TEM negatives with some scanners, we came to the conclusion that the minimum resolution we required was above 1.200 dpi. We too use Adobe Photoshop to control the scanner and to process the pictures. I hope this helps.
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Dear Isabel,
Just make your own standard. Adsorb colloidal gold particles to a poly-l-lysine coated formvar stabilised carbon support film on a TEM grid. Depending on what you want you can select Cu or an other material (carbon, nylon....) for the grid. Colloidal gold is available in ranges of sizes. Your calibration purpose needs large particles, which are up to about 40 nm commercially available (Aurion, Sigma, Amersham, Nanoprobe....).
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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Isabel Nogueira wrote: ======================================= I need a gold standard to calibrate an EDS analytical system attatched to a TEM.
I've contacted several suppliers but none of them have anything with gold (pure or in a compound).
Does anyone know where or how I can get a thin film standard of pure gold or of a compound including gold ? ======================================== SPI Supplies has up on our website (for address, see below) a number of different gold "standards" and "aids" for calibration purposes, including Prickley Gold on Copper and our Combined Test Specimen of small gold islands on a carbon film. If the data is taken near a grid bar you will be seeing data from both gold and copper. Micrographs accompany each product listing so you can see exactly what you are getting. We also offer thin microtomed sections of foils spanning the atomic number range for checking with other elements (e.g. besides gold).
Disclaimer: SPI Supplies is a manufacturer and distributor of these standards and calibration items as well as one-off custom made products for others.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
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The solution to your problem is DIC (differential interference contrast).
Regards, Michel
**************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
Anaspec South Africa, has an opening for an E.M. service engineer.
The position requires a person with tertiary electronics training or / = and E.M. experience on a technical basis. The job involves the maintenance and support of our customers E.M. = systems. This maintenance covers electronic, mechanical and P.C. repairs. We maintain a number of systems from various manufactures, Hitachi, Leo, = Topcon, Jeol, Philips EDAX, Oxford etc.
The support of our customers involves technical advice on E.M. matters = including operation, maintenance and special operating techniques. We also regularly present courses our offer in-house training.
Our customer base covers Southern Africa and a number of countries = internationally. Thus travel locally and internationally is part of the = job description. The applicant must therefor have a valid passport, a = reliable motor vehicle and be willing to travel when the need arises.
The remuneration package will depend on the experience of the applicant. = We offer commission on work done, Medical aid, life policies and car = insurance.
Please contact us on the numbers below if you are interested.
Luc Harmsen Anaspec, South Africa Technical support for E.M. operators, world wide. Anaspec-at-icon.co.za TEL: ++ 27 (0) 11 476 3455 FAX: ++ 27 (0) 11 476 7290=20
I am interested in acquiring a high resolution scanner. Is there one (e.g. Agfa DuoScan) that will scan both positive and negative originals AND provide crisp, high resolution scans? The DuoScan specs state that it will interface with UNIX workstations as well as PC and Mac.
Thanks for any and all feedback, Steve.
Steve Samuelsson, Ph.D. Procter & Gamble Pharmaceuticals, Inc. PO Box 8006 Mason, OH. 45040-8006 (513) 622-1753 office (513) 622-1752 lab (513) 622-1196 fax samuelsson.sj-at-pg.com
Dear Folks: I am also interested in negative scanners. If possible, could the respondees to Annette's question include approximate prices (in US$) and system requirements, and their experience with output quality from inkjet printers, such as the Epson stylus. Also, if you could give your opinion of the system for materials science applications, it sure would be appreciated.
Thank You in Advance,
Michael Coviello UT Arlington Materials Science Program Arlington, TX USA
I have been VERY pleased with the results from my UMAX 2000 scanner that does everything from prints to 35mm negatives. I have seen it recently advertised for as low as $2650. It has true 1000 X 2000 DPI resolution and will run from PC or Mac.
I have no financial interest in UMAX.
Mark A. Farmer Director, Ctr. Ultrastructural Research University of Georgia, Athens, GA 30602 (706)542-4080 Voice (706)542-4271 FAX farmer-at-emlab.cb.uga.edu
-- Constance Linville Dept. Neurobiol. & Anat. Wake Forest University School of Medicine Winston-Salem, NC 27157 email: clinvill-at-bgsm.edu
I am presetly involved in a study where I need to pick up serial pairs and they must stay connected! Sometimes they do great, sometimes they don't. I am very careful to have neat, smooth, and parallel block edges. I have also tried adjusting the water level. I need to find a technique that will increase the chances that when I cut, and pick up, the sections will stay together. I am collecting on formvar coated slot grids.
Hi Microscopists. This may interest histologists or others involved in biological samples. I want to ask for your input. The Massachusetts Office of Technical Assistance for Toxics Use Reduction is sponsoring a conference on Pollution Prevention Strategies for the Health Care Industry. We are issuing a call for papers (below). Some papers will be accepted for presentation and others for publication in a booklet to be distributed at the conference. We are looking for people who are using methods to reduce the use of formaldehyde, re-distilling their solvents, using dyes with fewer heavy metals or less toxicity, and other practices which reduce the toxicity of waste products or reduce the production of waste in general. Feel free to call or email me with questions, or if you have ideas for presentation or know of people from whom we might wish to solicit papers. Lara Sutherland, 617-727-3260. Email: Lara.Sutherland-at-state.ma.us.
CALL FOR PAPERS POLLUTION PREVENTION STRATEGIES FOR THE HEALTH CARE INDUSTRY
The Massachusetts Office of Technical Assistance for Toxics Use Reduction, inconjunction with the Massachusetts Water Resources Authority, the Massachusetts Department of Environmental Protection, the Medical, Academic and Scientific Community Organization, and the U.S. Environmental Protection Agency, will be hosting a conference on pollution prevention strategies for medical and dental facilities October 7, 1998, in Tyngsborough, MA. A booklet of short papers will be published, and a number of longer papers will be presented. Papers should be of a technical nature, focusing on true pollution prevention opportunities available to the health care industry. Conference focus will be the reduction of mercury, dioxin-forming incinerables, formaldehyde, solvents, and other materials of concern. Also covered will be recycling, reforms in purchasing and inventory, institutionalizing or improving environmental management systems, and overcoming barriers to organizational change. Vendor presentations of a strictly technical nature will be considered for conference booklet inclusion or session presentation. Abstracts (maximum length two pages) are due April 1, 1998, to Rick Reibstein, Office of Technical Assistance, Rm 2109, 100 Cambridge St, Boston, MA 02202. Inquiries may be directed to Mr. Reibstein, Lara Sutherland, or Scott Fortier, at 617-727-3260 x 688, 696 and 695 respectively. Vendors of products and/or services relating to pollution prevention in these facilities are invited to submit offers by June 1, 1998 to exhibit at a trade show associated with the conference. Offers should include sufficient information to characterize the technical and preventative nature of the product or service and should be directed to Joseph Paluzzi at the Office of Technical Assistance (address above, x693). This information is also available at OTA's website www.magnet.state.ma.us/ota/.
Is anyone out there recording images through the oculars on the microtome? If so, I would be interested in knowing your setup. I would like to record surface and polished defects after microtoming, through the oculars, without having to remove the sample thereby maintaining alignment/orientation. Can anyone recommend a digital camera which could fit onto the ocular?
Fred Hayes The Dow Chemical Co Analytical Sciences Midland, MI 517-839-4373
I have a question about flatbed scanners that maybe some of our high resolution friends might be able to answer if they have scanned Hi res images on a flatbed.
If you put a negative directly on the glass of a flatbed with a transparency adapter, you can get interference patterns in the scanned images. If you put them in a negative carrier to get them off of the glass you eliminate them. The thickness of my carrier is about 1 mm. The question that I have is does raising the negative off of the glass affect the focus of the scanned image? Is it blurred? Is there a depth of field associated with he optics of the scanner?
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
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Tobias Baskin wrote:
"Greetings, =09 Anyone out there know of a dealer who handles Aus Jena stock? As you may know, Aus Jena what was the East German branch of Zeiss was calle= d after the second world war and until the recent reunification of the companies (a few years after the country). I have a stand made by Jena th= at I'd like a part for, but Zeiss is no longer selling them. =09Thanks in advance for any leads," =09=09=09=09=09Tobias Baskin +++++++++++++++++++++++++
Tobias,
I know of a person who handled quite a bit of Jena scopes. His name is Ar= t Waldeck, Micro-Tech Instraments, 8031 North Academy Blvd., Colorado Spr= ings, CO 80902. His phone number is 719-495-8011 (v) and fax is 719-495-8= 008.
I think Art would be a very good place to start.
Also, the Seiler Instrument Co. in St. Louis, MO handled most? of the dis= tributorships in the country. They can be contacted at 314-968-2282.
by darkwing.uoregon.edu (8.8.8/8.8.8) with ESMTP id IAA05661; Fri, 16 Jan 1998 08:59:05 -0800 (PST) Message-ID: {34BF91D7.60ABAB0-at-darkwing.uoregon.edu}
Walck. Scott D. wrote:
} ... } I have a question about flatbed scanners that maybe some of our high } resolution friends might be able to answer if they have scanned Hi res } images on a flatbed. } } If you put a negative directly on the glass of a flatbed with a transparency } adapter, you can get interference patterns in the scanned images. If you } put them in a negative carrier to get them off of the glass you eliminate } them. The thickness of my carrier is about 1 mm. The question that I have } is does raising the negative off of the glass affect the focus of the } scanned image? Is it blurred? Is there a depth of field associated with he } optics of the scanner? } } ...
You ^should^ see moire patterns ... afterall, you are superimposing one orthogonal scan on another. I suspect your 1mm thick carrier is affecting focus and reducing the effect ... though its is an interesting remedy for the problem. If your resulting images are not as sharp as you'd like, you might try scanning your negatives at (eg) a 15degree angle, and then rotating them back to horizontal with your favorite image editor ... though this may also introduce an "un-sharpening" effect.
... hope this helps :o) cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility at the University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
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The Leafscan 45. Is in beta format and is about to be sold by Bremson Data Systems for about $28,000. You can reach them at 913-492-8900. I have no connection to them in any way; I am just ineterested in their system also. I believe they have upgraded the scanner quite a bit in terms of speed and color capability from the old Leafscan system.
Brad Storey Materials Scientist Argonne National Lab - West P.O. Box 2528 Idaho Falls, ID 83403 Ph. 208-533-7685 (office) Ph. 208-533-7439 (lab) Fax 208-533-7683 brad.storey-at-anl.gov
by welchlink.welch.jhu.edu (8.8.7/8.8.5) with SMTP id NAA03927; Fri, 16 Jan 1998 13:13:17 -0500 (EST)
Brandon, Try coating the top and bottom edges of the trimmed block with grid glue (double stick tape dissolved in choroform), the right concentration should make your serial sections stick.
} } we have a Zeiss EM902A. When using Ni-grids I observe that they stick to } } the specimenholder. Cu-grids do not. Both are formvar-coated. } } Has someboby regarded the same phenomenon? } } Does this mean that the specimenholder is magnetic? } Birgit-
I believe you have the Zeiss top-loading specimen holder with the screw-on cap, correct? Grids occasionally stick in the caps for several (non-magnetic) reasons. Formvar grids, especially, may have a tiny sticky tail that sticks in the cap. If you are seeing this only with the Ni grids, my guess is that the grids themselves are slightly different than your copper grids, perhaps ever so slightly larger, and that this, coupled with the formvar, makes them stick. Also, when this begins to happen, try carefully cleaning the tiny grooved seat in the cap under a microscope. Tiny bits of formvar may be wedged in the cap, enhancing the stickiness.
Aloha, Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Hello All, A couple of weeks ago I asked for opinions on the JEOL 'SemAfore' system they sell to take digital images off their SEM/STEMs. I received 14 replies:
2 were interested in the other replies; 1 saw the system at a demo, thought it was good but out of range of his budget; 1 said it wasn't as good as some other systems, 1 was about to install; he said it looked like it was good on image capture but pretty basic on image processing, 2 talked about digitising TEM images (I should have been clearer in my post - I'm already setting up a negative scanner for TEM); 2 users recommended other systems, i.e. semicaps and ImageSlave; and 6 tried to sell me some other system. Only one quoted a price.
I was a bit surprised not to find _anybody_ who has a working system of SemAfore. (Or perhaps it's not surprising, given the great diversity of systems out there, unless JEOL is pushing hard for this they won't sell many.) Or perhaps all the users are so pleased with their SemAfore system they spend all their time on the microscope and don't look at their e-mail. However, I think the main reason is that you get image capture automatically with a modern EDS system, so unless you're really strapped for cash you don't need to worry about it. That being the case, it seems to me that one should either go for the cheapest image capture system available that will do the job or get a decent EDX.
Looking forward to some more debate,
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ
The response was overwhelming! I'm trying to sort through the responses and try the suggestions until the problem is solved. Sorry I didn't reply personally to each one.
Does oxygen quench rhodamine fluorescence? I have read a few papers that used an oxygen scavenging system in their fluorescence experiments but I can't tell if that was to inhibit quenching or for some other reason particular to that system.
Bob
Dr. Robert R. Wise, Director UW-Oshkosh EM Facility Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
A colleague in Plant and Soil Science needs a method to detect dead plant cells in soybean root tissues. The tissues would be sectioned and then examined by light microscopy looking for numbers of dead versus live cells. Any stains available for this?
His address is: yluo-at-siu.edu
Many thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I am looking for an inexpensive alternative fixative for gross anatomical specimen storage. Presently, these specimens are stored in heat-sealed plastic bags containing 10% NBF. Some of these specimens are periodically removed from bag storage for teaching purposes.
I would appreciate any useful comments sent to the listserver, or telephoned / e-mailed to me directly.
Best regards,
Jerome Jasso Children's Hospital Medical Center of Akron, Ohio (330) 379-8279
Fred: I have not done this particular "stunt", but I know about ultramicrotomy and photomicrography and see no particular difficulty. You will require/find an adaptor tube with a thread that is often used for small movie cameras. They called C- mount and these are used with most digital cameras and any digital camera with a removable lens should do the job. It is important to use a very low power ocular in the C-mount tube (one or two times). Why such a low power photo ocular and several other matters relevant to digital microscopy are explained on our Pixera page in our online catalogue.
ProSciTech is an agent for digital cameras.
Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} Is anyone out there recording images through the oculars on the } microtome? If so, I would be interested in knowing your setup. I would } like to record surface and polished defects after microtoming, through } the oculars, without having to remove the sample thereby maintaining } alignment/orientation. Can anyone recommend a digital camera which } could fit onto the ocular? } } Fred Hayes } The Dow Chemical Co } Analytical Sciences } Midland, MI } 517-839-4373 } }
I know that this has been discussed in the past. But since I wasn't in the market at the time I didn't pay much attention. I would like to get some opinions on the Scion Corp and Data Translation frame grabbers. NIH image is setup for the Scion and Image Tool recommends the Data Translation board. I have been in contact with Scion and they are recommending their CG-7 board since it has oversampling capabilities its resolution may be greater. The Data translation board is the DT3155. Any one have anything to say about either system.
I would also like to get opinions on the Cohu 4910 series of monochrome CCD. Both of the above companies suggest that camera.
My main usage would be monochrome densitometry but the sections are small so I need good resolution without spending big bucks. A few thousand is all that I can come up with at the present time. This equipment would be on a Pentium PC. Any suggestions would be appreciated.
Zhengyu Wang wrote: ================================================ As you know, graphite is a good sample for AFM and STM while testing their resolution. Recently we want to buy some of graphite samples. But after I contacted with a few graphite companies, none of them would like to sell me a little amount of single crystal graphite sample. (or, they dont have single crystal graphite at all).
Does anyone know where can I buy a little amount of this stuff? Thanks so much and my best regards! ================================================ Actually what you really should be using is not what one might call (ordinary) graphite but "highly ordered pyrolytic graphite" or HOPG. This is a specialty item made in only a few manufacturing plants in the world. It is not something that is made by the traditional "graphite suppliers". It has very interesting and unique properties, one of the best known being its ability to undergo a micaceous type of cleavage, with new layers being exposed with a "Scotch tape stripping" process.
There seem to be subtle differences between the HOPG materials made from the two main manufacturing sources. We are convinced these are not "quality" differences, but material from the two different sources is indeed not exactly the same. However, each of the two manufacturing sources offers HOPG in several different grades as measured by what is called the "mosaic spread". It is an over simplification to say this is just a measure of disorientation, but to the first approximation, you could think of it in that way.
This is all explained on the SPI website in some detail. HOPG from both of the two major "sources" of HOPG is available, and for calibration purposes, we strongly recommend the highest quality (a.k.a. most expensive) grade be used, or putting it another way, the product grades with the very tightest "mosaic spread". For general research purposes, we recommend the next most highly ordered grade (some researchers prefer the more expensive grade even for general research work) and for classroom demonstrations and routine student use, the lowest priced quality should be considered. One important note here: As the mosaic spread becomes comes tighter and therefore more expensive, one also gets more "strippings" per "block". Therefore the price differential is less than at first it might appear.
Disclaimer: SPI Supplies offers HOPG to those doing SPM and thin film coatings research seeking to try new and innovative substrate materials.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I do not know the answer to this question. Would a few of you like to try and help a budding microscopist.
Nestor
Email: dbac-at-orbitworld.net Name: David Bacque
Question: Dear Sirs:
I am new to this, both the internet and microscopy so if my questions are inappropriate for this forum, please excuse me and discard.
My son Paul is 11 and received a good student microscopy for Christmas a year ago. He is an avid science student and is currently making slides of every feather he can find and is interested in his slides being more permanent. I have been trying to help him, but have not had much success. We started with the gum spirit in the Edmund Sci. microscope slide kit, but it takes forever to dry and seems to shrink as it dries causing terrible bubbles. I finally got some Cytoseal 60 locally and have had much more success however we still have problems with air bubbles creeping in from the edges days or weeks later. Are we using too much or not enough cytoseal? Do we need to seal the edges with something else? Is heating suggested to fully cure? How long should it take to dry? I thought this stuff was supposed to be fast, but it seems like days or weeks later the interior of the slide is still liquid. (or is it supposed to stay that way?) After drying should the specimen be soaked in solvent and if so which one?
Any other tips for making simple slides would be appreciated. I've been hunting the net and found lots of good information, but not the answers to these.
Thank you in advance and Paul also thanks you.
David Bacque Proud father and mentor of an avid young scientist
I received the following EMail from someone and thought it might be of interest to this List.
Ted Dunn Maui, Hawaii
{Tell your friends, tell everyone who uses E-Mail.. This is to inform you of a very important matter currently under review by the FCC. Your local telephone company has filed a proposal with the FCC to impose per minute charges for your internet service. They contend that your usage has or will hinder the operation of the telephone network.} E-Mail, in my opinion, will diminish if users were required to pay } additional per minute charges. The FCC has created an email box for your comments, responses must be received by February 13, 98. Send you comments to ""isp-at-fcc.gov"" tell them what you think. Every phone } company is in on this one, and they are trying to sneak it in just under the wire for litigation. Let everyone you know hear about this one. Get e-mail address to everyone you can think of. FCC E Mail address -- isp-at-fcc.gov
Guys, this is really important. If we have to pay for e-mail , the cost is going to skyrocket. It's about the only thing now that is cost-effective. Please make your opinions known to the FCC.} _____
I am looking for images of air pollution (dust particles, etc). I am an architecture student researching air contamination. I am not a scientist so I don't know under what type of microscopy discipline this would fall under.
Any leads or sources would be greatly appreciated!
} Email: dbac-at-orbitworld.net } Name: David Bacque } } My son Paul is 11 and received a good student microscopy for Christmas a } year ago. He is an avid science student and is currently making slides of } every feather he can find and is interested in his slides being more } permanent. } I have been trying to help him, but have not had much success. We started } with the gum spirit in the Edmund Sci. microscope slide kit, but it takes } forever to dry and seems to shrink as it dries causing terrible bubbles.
} Any other tips for making simple slides would be appreciated. I've been } hunting the net and found lots of good information, but not the answers to } these.
Paul -
Have you tried clear nail polish? Feathers are dry enough that it might work. You might need to thin it a bit with polish remover. Be careful when adding the corslip to lower it gently at an angle, rather than just dropping it flat; that is a sure way to get air bubbles. Please look at the Project MICRO bibliography (web address below). You'll find a lot of neat books listed, that have many ideas for things to do. Don't miss Devora Molitor's book, and the manual published by Usborne. Don't bother to print the booklist out if your dad is a member of MSA; members will be getting a copy in the mail later this month. [Listserver readers please note - the MICRO bibliography has been revised recently and is worth a return visit!]
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
This is a story that's been circulating for some time, but it's not accurate.
Here's what the FCC says:
"Q: Is the FCC considering allowing local phone companies to impose access charges on ISPs?
A: The FCC requested public comment in December 1996 on whether ISPs should pay current access charges, and more generally on how Internet and interstate information services that use local telephone networks should be treated. The Commission concluded on May 7, 1997 that ISPs should not be subject to interstate access charges. There is currently no open comment period on this issue.
Q: Does the FCC currently have an ongoing proceeding on Internet and interstate information services?
A: The FCC issued a Notice of Inquiry (NOI) in December 1996, at the same time as it asked for comment on whether ISPs should be subject to access charges. The NOI asked generally about how to create incentives for companies to make the most efficient use of the telephone network for Internet and other information services. The comment period for the NOI is closed, but the FCC has stated that it plans to issue a Notice of Proposed Rulemaking (NPRM) asking for comment on more specific proposals based on the responses to the NOI. {bold} The NPRM will consider actions other than imposition of per-minute access charges on ISPs {/bold} ."
Quoted from: Fact Sheet on The FCC, Internet Service Providers, and Access Charges
See the complete text at: http://www.fcc.gov/Bureaus/Common_Carrier/Factsheets/ispfact.html
Dave Harrison
Site Manager
JEOL USA, INC
At 04:36 PM 1/17/98 EST, you wrote:
} I received the following EMail from someone and thought it might be of
} interest to this List.
}
} Ted Dunn
} Maui, Hawaii
}
}
} { {Tell your friends, tell everyone who uses E-Mail.. This is to inform
} you of a very important matter currently under review by the FCC.
} Your local telephone company has filed a proposal with the FCC to
} impose per minute charges for your internet service. They contend that
} your usage has or will hinder the operation of the telephone
} network.} E-Mail, in my opinion, will diminish if users were required
} to pay } additional per minute charges. The FCC has created an email
} box for your comments, responses must be received by February 13, 98.
} Send you comments to ""isp-at-fcc.gov"" tell them what you think. Every
} phone } company is in on this one, and they are trying to sneak it
} in just under the wire for litigation. Let everyone you know hear
} about this one. Get e-mail address to everyone you can think of.
} FCC E Mail address -- isp-at-fcc.gov
}
} Guys, this is really important. If we have to pay for e-mail , the
} cost is going to skyrocket. It's about the only thing now that is
} cost-effective. Please make your opinions known to the FCC.}
} {Tell your friends, tell everyone who uses E-Mail.. This is to inform } you of a very important matter currently under review by the FCC. [snip] } Please make your opinions known to the FCC.}
I received this same e-mail on another mailing list which I am a member of, and it was revealed there that this is an urban myth. Here's the relevant portion of the reply offered on that list...
---------------------------Start Reply Here----------------------------------- As for this particular case, a quick check of the FCC web site reveals nothing of the sort, with that address being the comment address for something that is not all that related (in fact, it looks like the FCC is considering dropping interstate rates for ISPs...).
A quick check of my memory reveals that this particular urban legend of a data usage fee actually pre-dates the internet, and goes back to the days of BBS systems on 300 baud modems.
A check through the web turns up a small blurb on it at
} My son Paul is 11 and received a good student microscopy for Christmas a } year ago. He is an avid science student and is currently making slides of } every feather he can find and is interested in his slides being more } permanent. } I have been trying to help him, but have not had much success. We started } with the gum spirit in the Edmund Sci. microscope slide kit, but it takes } forever to dry and seems to shrink as it dries causing terrible bubbles. I } finally got some Cytoseal 60 locally and have had much more success however } we still have problems with air bubbles creeping in from the edges days or } weeks later. } Are we using too much or not enough cytoseal?
To try to describe an elephant while blindfolded, it sounds like your specimens are too thick. As the mountant dries the solvent dries out and thus the amount of mountant shrinks. This is accompanied by air spaces forming from the side. Depending on your specimens this may be very hard to overcome except by adding more mountant to the side of the cover slip at a corner of a "bubble" and letting it "wick" into the bubble.
} Do we need to seal the edges } with something else? Is heating suggested to fully cure? How long should } it take to dry? I thought this stuff was supposed to be fast, but it seems } like days or weeks later the interior of the slide is still liquid. (or is } it supposed to stay that way?)
Remember that the mountant dries for the outside in, and thus the center will take some time to dry. That, and many mounting media are made to stay soft in the center for various reasons.
After drying should the specimen be soaked in solvent and if so which one?
Depends upon the mounting medium and the specimen. "Most" mounting media have a Xylene base and xylene is a commonly used as a pre-mount solvent.
} Any other tips for making simple slides would be appreciated. I've been } hunting the net and found lots of good information, but not the answers to } these.
There are several tricks to making good slides, like the one Caroline Schooley mentioned of starting at an angle and slowly bringing the cover slip down.
I personally am a big user of Micro-miniature disposable wooden spatulas (commonly called flat toothpicks) as an aid in microscopy. They are great for slowly lowering the cover slip and other uses. Then you throw them away, no cleaning, very available and relatively inexpensive. And for those trying to quit smoking great to chomp on while thinking.
Pre-cut strips of filter paper (approx. 2 X 4 cm) are great to have on hand when you want to move, remove or clean liquid mounting medium from a slide.
Mounting medium cleans up a lot faster and easier when dry than when wet. Let it dry and then "chip off" with a No 11 scalpel.
Remember that Xylene, like all chemicals, should be treated with respect. Among other things, you don't want to breath too much of it, it is flammable, and can be problematic by causing discoloration to clothing and some work surfaces.
There is a mounting medium on the market that work on heat. (Commercial information provided by direct email if interested.) Heat it and it softens, let it cool to room temperature and it is set. The problem is that it is a PCB. While I do not worry about using it ,due to the extremely small quantities and shortness of contact, you may not desire to have your son use it.
A good safety feature is a plastic jar with a mouth wide enough to throw the to be disposed of slides and other glassware in. Keep it right where you work. This will save you a lot of broken glass to clean up and hopefully some cuts.
On making "good slides" there is nothing that beats, practice, practice, and more practice. That and cleaning up a lot of mountant drops and smears and throwing away a lot of your first experiments.
Good luck and Shalom from Jerusalem Azriel
+++++++++++++++++++++++++++++++++++++++++++++++++++++ Azriel Gorski, Head Fibers and Polymers Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem, ISRAEL
azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739
What lies behind us and what lies before us are tiny matters compared to what lies within us. Ralph Waldo Emerson ++++++++++++++++++++++++++++++++++++++++++++++++++++
} There are several tricks to making good slides, like the one Caroline } Schooley mentioned of starting at an angle and slowly bringing the cover } slip down.
{snip of balance} Another tip that Azriel uses, I'm sure, and just forgot to mention is weighting the cover slip. A bit of lead, like a fishing sinker or a bullet (if you happen to have any handy!) works well. When I worked in a crime lab I kept a dozen or so .45 ACP cartridges at my sample prep bench and weighted all cover slips put over solvent-based mounts. These seemed to be a good weight for 18-22 mm square cover slips. I'd use two if using 22X40 cover slips. (I didn't use them on my other then-current practice of mounting in Aroclor.) You may need to leave the weights in place for several days, depending on how long it takes for the prep to become relatively solid.
Best of luck! -- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
1- Linkam of UK {A HREF="http://www.linkam.co.uk/"} Linkam Scientific Instruments Ltd Home Page {/A} makes an observation cell that can be used with SAXS (or light analysis) ; that is temperature-controlled to do cooling- heating. Would any one know of a US-based company that carries the similar equipment?
2- Would any one know of companies that use flow cells ,(fabricate/customize to fit client' s needs) either as a cell for UV-Vis &/or for Light microscopy (dimensions ~ 1 sq.in.) ?
THANKS for the reply given to previous inquiry by : Subj: Re: Videomicroscopy and In-situ crystal growth
id sma018090; Mon, 19 Jan 98 15:31:47 +0100 Received: from [193.74.131.15] by rxu01 with SMTP (1.37.109.4/16.2) id AA14924; Mon, 19 Jan 98 15:18:39 +0100
Thanks all for the replies to my query regarding the existence of a good atlas of virus morphology. For those interested, here is the summary (some lines deleted).
Interestingly, there does not seem to exist a recent book (editors please take notice!) but there is obviously a lot of data online. Hence, happy browsing!
Best regards, Michel
************************************************************ I tried some WWW searches, might try some of the following site, it looks= good:
Lou Ann ************************************************************ Perhaps this is not what you are specifically looking for, but our group has imaged virus in the cryo stage of our Hitachi S-900 FESEM. I believe Ya Chen, our SEM coordinator, has included some of the virus work in our web page, found in my signature below.
Sincerely,
Colleen Lavin Integrated Microscopy Resource 1675 Observatory Dr. Madison, WI 53706 608-263-8481 voice 608-265-4076 fax lavin-at-calshp.cals.wisc.edu http://www.bocklabs.wisc.edu/imr/home2.htm ************************************************************ The Atlas of Virus Diagrams, Hans-W. Ackermann and Laurent Berthiaume, CRC press,Inc., 1995 seems to be up to date with the Pox family. There are no micrographs only diagrams. Andy
Andrea Weisberg NIH/NIAID/LVD Build. 4/ Rm. 210 4 Center Drive Bethesda, Md 20892-0445 (301) 435-1977 Office (301) 480 1147 Fax aweisberg-at-nih.gov ************************************************************ Yes, there is a problem in getting nice EM pictures using cryo-EM. Most EMs are from the old "garde", they are still pretty good and I have been trying during the last 2 years to "unearth" some of the originals to scan them in and to put them on the Web. I have started with a virus "gallery", but not all of them are good. My principle interest is to have a good image of each virus genus at least. Have a look at http://life.anu.edu.au/viruses/Images/ or at http://www.tulane.edu/~dmsander/Big_Virology/BVHomePage.html. The later one is using a lot of my images and my virus system, but that is ok.
If you find nice images, please let me know.
Warm greetings from down-under Cornelia
Dr Cornelia B=FCchen-Osmond Bioinformatics Group Research School of Biological Sciences Phone: 61 (02) 6249-4842 The Institute of Advanced Studies Fax: 61 (02) 6249-4437 The Australian National University E-mail buchen-at-rsbs.anu.edu.au GPO Box 475, Canberra, ACT 2601, Australia http://life.anu.edu.au/viruses/welcome.htm
=20
**************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be=20 Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals =20 Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
I am about to purchase some goat-anti-rabbit 5 and 10 nm colloidal gold = conjugates for immunolabelling, and was wondering if anyone had any = comments about specific vendors re: quality, uniformity, labeling = intensity, stability, etc.
Please send comments directly to me, and I will compile a list and post = it to the listserver.
Thanks in advance!
Craig R. Lending Department of Biological Sciences SUNY Brockport Brockport, NY 14420
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I have received several questions about the Leafscan 45. I don't have one and I have no financial interest in the company. Call them, not me. I have seen the scanner in action at ANL and LBL. Both places are very picky about image quality and they are happy with the scanner (last time I checked). It is not a drum scanner and yes it is about 28k it used to be about 17k. It is a lot of $, but it isn't a cheap multipurpose flatbed desk scanner.
Call the company not me (i don't have time to respond to everyone). I guess the number of responses I got is a testament to the size of this listserver.
Brad Storey Materials Scientist Argonne National Lab - West P.O. Box 2528 Idaho Falls, ID 83403 Ph. 208-533-7685 (office) Ph. 208-533-7439 (lab) Fax 208-533-7863 brad.storey-at-anl.gov
id MAA06809 for {microscopy-at-msa.microscopy.com} (from jcraig-at-unbc.ca); Mon, 19 Jan 1998 12:25:58 -0800 (PST) Received: by gpu.unbc.ca (8.6.12/UNBC-2.6C) id UAA18351 (from jcraig-at-unbc.ca); Mon, 19 Jan 1998 20:25:58 GMT
Hi all,
I'm resending this so I appologize if you've already received it. Thank you to everyone who replied to my Edax/Windows problem and SEM software problem. The number of replies was astonishing and very helpful.
We have a benchtop March Instruments parallel plate etcher. We are currently using it to etch off layers of dielectric between metal layers on our semiconductors. We are interested to find out if anyone has an etcher for a similar purpose and what instrument they are using. We have not been able to find any small etchers other than the March Inst. model. Thanks for your input. Mark Windland Analytical Services Solid State Electronics Center Honeywell Minneapolis, Minnesota 612-954-2845 fax 612-954-2040 e-mail: windland-at-ssec.honeywell.com
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1/19/98 2:27 PM Frontiers Meeting venders info
To all electron microscopy venders:
As a reminder The Seventh Conference of Electron Microscopy in Materials Science is being held at Kloster Irsee Irsee, Germany. The dates for the meeting are April 19-24th 1998. The vender session will once again be a single day table top exhibit only. The tentative date for the vender session will be Monday the 20th and will be an evening session following dinner. We will most likely be limiting the number of exhibitors to #197# 15. The cost is $600 US. Please contact Joe Mayer at JMAYER-at-vaxww1.mpi-stuttgart.mpg.de for reservations and details. There will also be advertising space in the preprinted abstracts. If you cannot attend the meeting but would like your product information to be available please contact me at the below address. You can also get more details about the meeting at http://multiscale.llnl.gov/femms98/default.html.
Thanks,
Mr. Mark A. Wall L-350 Chemistry & Materials Science Dept. Lawrence Livermore National Laboratory 7000 East Ave Livermore, CA 94550 USA ph. 510 423-7162 fx. 510 422-6892 e-mail. mark.wall-at-quickmail.llnl.gov
by ereapp.erenj.com (8.8.5/8.8.5) id TAA05006; Mon, 19 Jan 1998 19:09:14 -0400 Received: from eredns.erenj.com(159.70.1.252) by ereapp.erenj.com via smap (V2.0) id xma005002; Mon, 19 Jan 98 18:08:54 -0500 Received: from clmail.erenj.com (clmail.erenj.com [159.70.1.248]) by eredns.erenj.com (8.8.5/8.8.5) with ESMTP id TAA03342; Mon, 19 Jan 1998 19:08:53 -0400 Received: from clnmjluton1.erenj.com (clnmjluton.erenj.com [159.70.16.212]) by clmail.erenj.com (8.8.5/8.8.5) with SMTP id SAA18712; Mon, 19 Jan 1998 18:08:52 -0500 (EST)
Post Doctoral Position TEM in Physical Metallurgy Corporate Research Laboratory, Exxon Research & Engineering Co. Route 22E Clinton Township, Annandale, NJ 08801
A post doctoral position is available immediately in the Advanced Materials Section of the Corporate Research Laboratory, Exxon Research and Engineering Company.
The position requires demonstrated expertise and experience in the analytical and high resolution transmission electron microscopy characterization of microstructures and fine precipitates in ferrous and other alloy systems. A broad physical metallurgy background, including structure-property relations, phase transformations, and phase stability is also required. The employee will be expected to contribute to the development of new high strength steels tailored for applications in oil and gas industries. In a multidisciplinary and cooperative research environment, the employee should be able to work effectively in a team and should have good interpersonal and communication skills.
Exxon's Corporate Research Laboratory is located in scenic, rural western New Jersey, about an hour west of New York City and 45 minutes northwest of Princeton. The laboratory performs basic and applied research in support of Exxon Corporation's worldwide scientific, technological, and business needs. We offer an excellent working environment.
Qualified applicants should send their resume to the above address or to fax 908-730-355 before February 1, 1998. Enquiries can be made by calling Dr. J.K.Chen at Tel: 908-730-2756 or E-mail: jkchen-at-erenj.com
SALZBURG, 20th of Jan, 1998, local time: 01.10 a.m.
Dear Michel, dear Dr. Deschuyteneer, unfortunately I'm a little bit too late with my reply. But maybe it would be interesting to know of the following books: 1) Erskine L. PALMER & Mary Lane MARTIN (Eds): An Atlas of Mammalian Viruses CRC PRESS, BOCA RATON, FL, C: 1982, 2nd ed.: 1985 ISBN 0-8493-6628-3 (154 pages incl. subject index, and a lot of B/W-micrographs, good quality, "the elder bestseller") 2) David O. WHITE & Frank J. FENNER (Eds): Medical Virology, 4th Ed. ACADEMIC PRESS, San Diego, N.Y., etc. 1994 ISBN: 0-12-746642-8 (603 pages incl. subject index, but some less B/W micrographs than above compared to the overall pages; good quality) 3) F. FENNER & A. GIBBS (Eds): Portraits of Viruses: A History of Virology KARGER, Basel (Switzerland), Munich, etc., 1988 ISBN: 3-8055-4819-2 (344 pages, incl. subj. index; only 45 figures B/W, not all correspond to virus figures; and 9 tables)
Hope to add some new to the list Best regards and "Good night, America"
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
At 03:41 PM 1/19/98 -0600, Mark Windland asked: } We have a benchtop March Instruments parallel plate etcher. We are currently } using it to etch off layers of dielectric between metal layers on our } semiconductors. We are interested to find out if anyone has an etcher for a } similar purpose... {snip}
VG Microtech, in the UK, manufactures a plasma etcher for this application. We (Energy Beam Sciences, Inc.) distribute this instrument in the United States. I would be happy to send product or applications information to anyone who contacts me directly. Information is also available, on-line, at our WWW site: http://www.ebsciences.com
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
May 21-23 and May 25-27, 1998 North Carolina State University Raleigh, North Carolina, USA
and
June 15-18, 1998 Danish Technological Institute Taastrup, Denmark
This highly regarded hands-on course taught by expert faculty has been presented annually for more than 15 years. It deals with all phases of quantitative and computer-assisted imaging from acquisition and processing through measurement and stereological interpretation. Attendees receive The Image Processing Handbook plus a CD-ROM containing images, algorithms (Photoshop-compatible for Mac and Windows) and an extensive on-line tutorial and course notes on stereology and statistical analysis. The course is appropriate for scientists, technicians and administrators using or intending to use these techniques. Attendees typically come from materials science, geology, biological and medical sciences, pharmaceuticals, food science, industrial quality control, remote sensing, and other disciplines. You are encouraged to bring your own images for the hands-on lab sessions.
For detailed information and registration contact Alice Warren, Dept. of Continuing and Professional Education, N. C. State University, Raleigh, NC 27695-7401, 919-515-8171, fax 919-515-7614, email: kelly_mcdowell-at-ncsu.edu
Information is available on-line at the following sites:
I am working with isolated Golgi membranes deposited on 300 mesh nickel grids covered with 1% formvar (+/- carbon film), then fixed and stained with a negative contraster. The problem is that the membranes are not strongly attached to the film and due to the air drying they glide and cluster togheter creating a big problem in the evaluation of the results.
Dr. Roberto Weigert Consorzio Mario Negri Sud Department of Cell Biology and Oncology Molecular Neurobiology Laboratory Via Nazionale S.M. Imbaro, 66030 Chieti Italy Phone: 0039-872-570-354 Fax: 0039-872-578-240
Post Doctoral Position TEM in Physical Metallurgy Corporate Research Laboratory, Exxon Research & Engineering Co. Route 22E Clinton Township, Annandale, NJ 08801
A post doctoral position is available immediately in the Advanced Materials Section of the Corporate Research Laboratory, Exxon Research and Engineering Company.
The position requires demonstrated expertise and experience in the analytical and high resolution transmission electron microscopy characterization of microstructures and fine precipitates in ferrous and other alloy systems. A broad physical metallurgy background, including structure-property relations, phase transformations, and phase stability is also required. The employee will be expected to contribute to the development of new high strength steels tailored for applications in oil and gas industries. In a multidisciplinary and cooperative research environment, the employee should be able to work effectively in a team and should have good interpersonal and communication skills.
Exxon's Corporate Research Laboratory is located in scenic, rural western New Jersey, about an hour west of New York City and 45 minutes northwest of Princeton. The laboratory performs basic and applied research in support of Exxon Corporation's worldwide scientific, technological, and business needs. We offer an excellent working environment.
Qualified applicants should send their resume to the above address or to fax 908-730-3355 before February 1, 1998. Enquiries can be made by calling Dr. J.K.Chen at Tel: 908-730-2756 or E-mail: jkchen-at-erenj.com
Exxon is an Equal Opportunity Employer M/F/H/V
-------------------------------------------------------------- J.K.Chen, PhD | Postdoctoral Fellow Exxon Research & Engineering Co. | jkchen-at-erenj.com Route 22 E, Clinton Township | Tel: +1-908-730-2756 Annandale, NJ 08801 | Fax: +1-908-730-3355
Dear Community-- We have a Kinney High Vacuum Evaporator (Model SC-2), and I'm trying to locate an OFEC Hi Vacuum Globe Valve, size 1" (sweat type, straight through). This is for the Backing circuit--the seat is OK, but the brass bellows on the valve has given up. I contacted Kinney (now part of Tuthill Corp) and they know nothing.... jptmvl-at-mailbox.syr.edu thanx
Responding to the message of {v03007807b0ea63f8760a-at-[206.69.208.21]} from David L Johnson {jptmvl-at-mailbox.syr.edu} :
} Dear Community-- } We have a Kinney High Vacuum Evaporator (Model SC-2), and I'm trying to } locate an OFEC Hi Vacuum Globe Valve, size 1" (sweat type, straight } through). This is for the Backing circuit--the seat is OK, but the } brass bellows on the valve has given up. I contacted Kinney (now part of } Tuthill Corp) and they know nothing.... } jptmvl-at-mailbox.syr.edu } thanx
David,
I have a Kinney evaporator model KSE-2A-M. A few decades back the Kinney line was taken over by Sharon Vacuum, Sharon Machine Co., Inc, 69 Falmouth Ave., Brockton, Mass. 02401. (617)588-2323.
I last ordered parts from them in 1980 (!), and my machine has been working like a charm ever since. I have no idea if they are still in business, but give it a try.
Good luck,
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Fellow histoneters, I am patiently waiting for the Leica rep to come and remove the thermal unit from my EM auto processor! I was informed, no loaners available! If you have a service contract, a loaner is shipped out immediately. Which is great when they have a loaner. Except for the fact that the processors weigh alot, or it seems like it especially when lifting from floor. I also have to use the same box to ship "sick processor" to them. That means, lifting, packing, etc. Twice (within span of 2 months) I have walked into the em lab, with the processing vial melted inside the thermal unit, solution evaporated, of course (temps hi enough to melt vial) thank God it was not Acetone, but 50% ethanol. The screen is blank. I, nor any one else in immediate surrounding area, heard any warning sound or beeping sound. I am not sure if anyone else is having this very serious problem. If so, did it help to remove the thermal unit? Teresa
I am in the market for a flatbed scanner and would like to here your opinion regarding choices of flatbed scanners (such as specific brand, model, cost,...etc). The scanner will be used for digitizing the EM micrographs or images which are to be either archived or used for quantitative image analysis.
I know that this is an old subject for this group, can someone help me find a summary list of the scanners that were discussed previously by this group? Lastly, Is there any new and better model available since the subject was last discussed? Your information will be greatly appreciated.
Anyone out there using UNICRYL embedding resin as an alternative to LR-WHITE for immunogold labelling?
We need to thin-section through an embedded cell monolayer grown on glass coverslip and have been unsuccessful in
1) getting the resin to polymerize properly at 4C with UV and
2) Removing the embedded layer from glass (HF destroys (softens) the resin rendering it useless for sectioning). These cells will only adhere to either glass or polystyrene.
Any feedback with useful tips or your opinions on the use of these 2 resins would be appreciated.
If you are digitizing prints, the requirements are probably not that great. To get a 2000x1600 pixel image from a 4x5 inch print only requires 400 dpi of resolution. And depending on your application, that may be more resolution than you need. If you are working from negatives or smaller formats, then you would have more stringent requirements.
We have used the HP series of scanners with good success. We also purchased a cheaper model and had some regrets. Even our old HP IIcx was quite a bit faster than the new one, and the HP user interface was more intuitive and well behaved. I would strongly look at those two factors and insist on a demo before making a choice.
At 02:01 PM 1/20/98 -0500, you wrote: } I am in the market for a flatbed scanner and would like to here your } opinion regarding choices of } flatbed scanners (such as specific brand, model, cost,...etc). The scanner } will be used for digitizing the EM micrographs or images which are to be } either archived or used for quantitative image analysis. } } I know that this is an old subject for this group, can someone help } me find a summary list of the scanners that were discussed previously by } this group? Lastly, Is there any new and better model available since the } subject was last discussed? Your information will be greatly appreciated. } } Tsuey-Chen Long } WL Gore and Associates ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
I would like to address your comments regarding the most reliable cold stage for an SEM. I appreciate your loyalty to your Oxford equipment but I wonder if you have had the opportunity to operate the Emitech cold stage. I'm afraid that we cannot help but take offense at your comment that we "pale into insignificance". The Emitech cold stage has been on the market for serveral years with a great deal of success and our customers seem very pleased with its ease of operation and reliability.
We understand that it is not appropriate for vendors to use the ListServer to "toot our own horn" but felt that we could not take this slam against our equipment, as well as other manufacturer's equipment, without some response.
Regards,
John Fitzpatrick President and CEO Emitech U.S.A., Inc. (281)893-2067
TO: microscopy-at-MSA.Microscopy.com SUBJECT: Seeking MT-2 Vise-type specimen holders
We are looking for new or used vise-type / flat specimen holders/chucks for the Sorval MT-2 series of ultramicrotomes. If anyone has any stuck away in a drawer that they are no longer using we would be very interested in working out some kind of a deal. If anyone knows of a comercial source for buying new/used vise-type holders we also be very grateful.
Please respond directly to me at: carney-at-marshall.edu
Michelle D. Carney Zill Lab Marshall University - School of Medicine Department of Anatomy, Cell and Neurobiology 1542 Spring Valley Drive Huntington, WV 25755 Phone: (304) 696-7384 fax (304) 696-7290 E-Mail: carney-at-marshall.edu OR carney-at-zoomnet.net
Ronald M. Anderson (1-914-892-2225) wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I'm posting this as a favor to the individual named below. } She asked me for info on possible SEM courses and I was not able } to help. Excerpts from her notes are below. She has already contacted } Lehigh regarding their courses. } } Marisa Ahmed wrote: } } I work in the lab of an engineering company in Kanata, ON and want } information/training on sample preparation and delayering } of IC's (for SEM mostly) for failure analysis-type work. } If, as Prof Lyman suggested, you could help me locate such a possible } course or resource I would appreciate it. } } ------------- } And then a day later after I told her my course was TEM prep... } ------------- } } You are correct, I am not specifically looking for a course for TEM } prep (especially since we don't have a TEM here), although I would be } interested in knowing what was included in your course and where it is } being taught for future reference. I am actually hunting for something } to brush up my SEM sample prep (for topo views and delayering of IC's } mostly, not so much cross-sectioning). I also want to learn } something about the replica-stripping technique that I believe is used } for STEM and some SEMwork for radioactive samples. } } If you could post my query I would much appreciate it. Respondants can } reach me at mahmad-at-semiconductor.com } } Thank-you again, } } Marisa } } ((Sounds interesting. Please also copy me on responses. Ron A.)) Dear Marisa,
In response to Ron Anderson's posting on the listserver, I believe we, Allied High Tech Products, Inc., have what you are looking for. We offer courses for SEM sample preparation and delayering of IC's. In addition, we offer courses, not specifically for, but include: PRE-FIB Thinning for TEM, Backside Sample Prep for Package Devices that includes cross sectioning C4 devices, TEM Sample Preparation, and basic metallography.
We also manufacture equipment that includes our NEWEST Polishing Machine, "TechPrep8" and "MetPrep8", our Variable Speed Sectioning Saw, "TechCut", and a Positioning Device designed for precision lapping and polishing of microelectronic devices and die, "MultiPrep". The positioning device features a universal mount for using several different types of sample fixtures.
If you would like more information, please contact us at 1-800-675-1118, and I will be glad to discuss your specific interest. I can also mail you some literature on our products and equipment if you give me your address and phone number. Our WebSite contains photo's of our new equipment with the exception of the TechCut, which will be posted soon.
I look forward to hearing from you.
Sincerely,
Gary Liechty Product Application Specialist Allied High Tech Products, Inc. 2376 E. Pacifica Pl. Rancho Dominguez, Ca. 90220 1-800-675-1118 1-310-762-6808 Fax www.AlliedHighTech.com
Products for Materiallographic, SEM and TEM Sample Preparation
hi all, I am seeking information, the current image capture card has died and I am looking for a replacement.
What is the colective wisdom on image capture cards. The pro/cons of different makes.
Stuart --------------------------------------------------------------------------- Stuart G. McClure, || Post small : P.Bag #2, Glen Osmond, CSIRO Land and Water, || S.A. , AUSTRALIA, 5064. Adelaide Laboratories, || Post large : Waite Rd, Urrbrae, S.A., Australia || S.A., AUSTRALIA, 5064. --------------------------------------------------------------------------- Phone: (08) 8303-8484 International replace 08 with 61-8 Fax: (08) 8303-8550 Email: Stuart.McClure-at-adl.clw.csiro.au --------------------------------------------------------------------------- "There is a special providence in the fall of a sparrow; If it be now, it is not to come; If it be not to come, it will be now; If it be not now, yet it will come: The readiness is all." Hamlet ---------------------------------------------------------------------------
Dear list, I was very fast to delete summary of (or detailed replay to) request for ion millers producers some time ago. Would someone not so quick forward me that message (or any additional oppinions) off-list.
Hi. I am very interested in doing a thorough quantitative study on the objective lens of the TEM. I am just getting started on my study and am putting together a proposal on what I want to cover in my study. I would like as many pointers and suggestions from anyone familiar in this field. I am also interested in the pos- siblity of getting my hand on a spare/retired piece or parts of the objective lens. I would appreciate any suggestions on this subject as well. As I have already mentioned, I am just getting started. Any suggestions would be very very helpful. Thank you.
William Chung willyc-at-jhu.edu ______________________________William Chung____________________________
Johns Hopkins University, Biomedical Engineering
3003 North Charles Street Apt #316C Baltimore, MD, 21218 USA
I am planning to give a graduate course on Applications of scanning electron microscopy in materials science and asking if there is any literature, textbook that can be recommended.
Thank you for your message about Low Temperature Microscopy and the equipment that is available. I have indeed used the Emitech Equipment and the equipment made by other vendors.I have over the past thirty years built cold stages for a number of different microscopes. I stick by what I said but if you would like me to give you a frank assessment of the Emitech equipment, please write to me.
Patrick Echlin Director, Multi-Imaging Centre School of Biological Sciences University of Cambridge CAmbridge CB2 3EA United Kingdom
equipment stagesOn Tue, 20 Jan 1998 Emitech-at-ix.netcom.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Dr. Echlin: } } I would like to address your comments regarding the most reliable cold stage for an SEM. I } appreciate your loyalty to your Oxford equipment but I wonder if you have had the opportunity } to operate the Emitech cold stage. I'm afraid that we cannot help but take offense at your } comment that we "pale into insignificance". The Emitech cold stage has been on the market for } serveral years with a great deal of success and our customers seem very pleased with its ease } of operation and reliability. } } We understand that it is not appropriate for vendors to use the ListServer to "toot our own } horn" but felt that we could not take this slam against our equipment, as well as other } manufacturer's equipment, without some response. } } Regards, } } John Fitzpatrick } President and CEO } Emitech U.S.A., Inc. } (281)893-2067 } }
I suggest that you coat your formvar grids with an antibody directed against the Golgi membranes for half an hour, wash and afterwards deposit your membranes. You can also glow discharge before the deposit. Hope this helps, Daniele -- =
************************************************************** * Dani=E8le SPEHNER, CJF 94-03 INSERM * * Etablissement de Transfusion Sanguine de STRASBOURG * * 10 rue Spielmann, B.P. 36- 67065 Strasbourg-Cedex, FRANCE. * * Tel : (33) 03 88 21 25 25 - Fax : (33) 03 88 21 25 21 * **************************************************************
With reference to a suitable text book for the SEM of Material I would suggest "Scanning Electron Microscopy and X-ray Microanalysis" written by Goldstein et al Plenum Press New York 1992 (2nd Edition)ISBN 0-306-441175-6
Patrick Echlin Director, Multi-Imaging Centre School of Biological Science University of Cambridge Cambridge CB2 3EA UK
On Wed, 21 Jan 1998, svepet wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am planning to give a graduate course on Applications of scanning electron } microscopy in materials science and asking if there is any literature, } textbook that can be recommended. } } Sten Johansson } }
} Anyone out there using UNICRYL embedding resin as an alternative to } LR-WHITE for immunogold labelling? } } We need to thin-section through an embedded cell monolayer grown on } glass coverslip and have been unsuccessful in } } 1) getting the resin to polymerize properly at 4C with UV and } } 2) Removing the embedded layer from glass (HF destroys (softens) the } resin rendering it useless for sectioning). These cells will only } adhere to either glass or polystyrene. } } Any feedback with useful tips or your opinions on the use of these 2 } resins would be appreciated. } } Thanks, Karen Rethoret } York University } }
Dear Karen,
We have been usigng Unicryl for post-embedding reactions on animal tissues.
- If the resin doesn't cure, you may try to place the UV-lamp closer to your object.
- We found that some specimen holders (e.g. microtiter-plates) absorb UV-radiation. Therefore it might help to place the lamp in a way that puts nothing between your specimen and the UV-source (irradiation from above or below...)
- Your UV-lamp might be old! Our experience, based on information from a friend with a fish-basin, is that the UV-part of the lamp decreases with longer usage.
- Oxygen also keeps the resin from curing properly. You could put a lid on or try a nitrogen-atmosphere.
Hope this helps, Monika kroez-at-patho.vetmed.uni-muenchen.de
I started working with an AFM recently. In connection with my topic I need to be able to produce small droplets (approximately 100 nm in height and a few micrometers in diameter) on a ceramic surface. These droplets have to stick well, since they have to undergo a variety of abrasive procedures. Moreover am I seeking for a substance which - does not shrink - does not swell in water and - does not react with acids. Any advice on a suitable substance and how to apply it is highly appreciated! Thanks Manuela
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Memo : microtomy of microcrystals 21-01-1998 15:01
Dear all,
we are looking for recent papers, review articles, books or proceedings on microtomy of solid state crystals. The type of crystals are of micron scale and dispersed in a plastic block. As we want to do high resolution electron microscopy we need to have very thin sections of the order of 10 to 50 nm, if possible.
Places in Europe where we could get some training could also be interesting.
Thanks for your time,
Nick Schryvers
Dr. Dominique Schryvers University of Antwerp, RUCA - EMAT Groenenborgerlaan 171, B-2020 Antwerpen (Belgium) Tel: 32-3-2180247 Fax: 32-3-2180257 e-mail: schryver-at-ruca.ua.ac.be homepage: http://www.ruca.ua.ac.be/~EMAT/Schryvers.html
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Hello list members
I wonder if anyone could help me get in contact with Ultraclone Limited. They are the named producer of a particular antibody in an experiment I am trying to duplicate.
Ultraclone Limited Rossiters Farm House Wellow, Isle of Wight PO 401-OTE, UK
I have contacted them by snail-mail and had no response. Can anyone provide an e-mail address or telephone or fax number?
Any help or information would be greatly appreciated.
I have used both Umax and Microtek and have been happy with each. For evaluation reports you might try: http:\\www.zdnet.com This is the entry point. Couldn't get the exact (further into site) address while at work... Think java was crashing the NS 2.0 we run at work :(
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
Home woody.white-at-worldnet.att.net http://www.geocities.com/capecanaveral/3722
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I am in the market for a flatbed scanner and would like to here your opinion regarding choices of flatbed scanners (such as specific brand, model, cost,...etc). The scanner will be used for digitizing the EM micrographs or images which are to be either archived or used for quantitative image analysis.
I know that this is an old subject for this group, can someone help me find a summary list of the scanners that were discussed previously by this group? Lastly, Is there any new and better model available since the subject was last discussed? Your information will be greatly appreciated.
release (PO205-101c) ID# 0-42511U8000L8000S0) with SMTP id AAA180; Wed, 21 Jan 1998 10:29:01 -0600 X-Sender: tflore-at-pop3.lsumc.edu Message-Id: {v01510102b0eb8f3dee0d-at-[155.58.72.72]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: histonet-at-pathology.swmed.edu
A histotechnologist phoned me with this question and does not have e-mail available so I thought I would place this for any help. A surgeon asked pathologist in her group if a control "nocardia" was being used when doing Fite, Lepro or Ziel stain. The tech has phoned surrounding hospitals and no one seems to know or have any such control. Is there anyone who has used or worked with this type of control? Thanxs, in advance, Teresa
We finally (thank god!) replaced our 10MByte Bernoulli disk drives on our Kevex 8000 system with SyQuest drives. Kevex supplied us with a program that converts spectra and images to .tiff files. Has anyone had experience with sending these files to a Windows-based pc? How is it done? Do I need Kermit or is there a simpler way?
Use a polypropylene or polyetylene substrate. I work with the growth of pollen tubes in culture medium and I had excellent results using these plastic materials as a subtrate for the medium with Spurr's resin. I am working with Unicryl and I haven't any problems during the polymerization under 4 C with UV.
Rinaldo Pires dos Santos Dept. of Botany Universidade federal do Rio Grande do Sul Brazil e-mail: rinaldop-at-botanica.ufrgs.br
You will need some sort of communication program. Kermit works as well as anything for serial lines to another platform. I seem to recall that Kevex had some utility of their own, but we never ordered it for our Kevex Delta. We put an Ethernet card in ours and used FTP software to ship files back and forth. Whereas Kermit approaches 1 kbyte per second at 9600 baud, FTP was a couple of orders of magnitude faster. But then there was the extra cost of the card and software. And how many files do you have to ship anyway? Shipping a batch overnight works pretty well.
As far as the utility to convert things to TIFF, we wrote our own. When doing so, I found that not all applications see all TIFF files the same way. Some features we built in were not recognized by some programs, or those programs insisted on a different tag for the purpose. So try the files with another program if it doesn't work with the first program you try.
On the side, how big a Syquest cartridge does your system handle? I thought the DEC architecture was limited to 32 MBytes per disk chunk unless you got some special drivers. But even 32 MBytes was a lot of space in those days.
At 11:41 AM 1/21/98 -0600, you wrote: } We finally (thank god!) replaced our 10MByte Bernoulli disk drives on } our Kevex 8000 system with SyQuest drives. Kevex supplied us with a } program that converts spectra and images to .tiff files. Has anyone had } experience with sending these files to a Windows-based pc? How is it } done? Do I need Kermit or is there a simpler way? ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
we are currently trying to establish a specific stain for type II pneumocytes in the mammalian lung. Has anybody experience with alkaline phosphatase staining of these cells and could give some practical details? Or are there any other suggestions for stains/markers for type II pneumocytes? Thanks in advance,
Matthias Matthias Ochs, M.D. Dept. of Anatomy Div. of Electron Microscopy University of Goettingen Kreuzbergring 36 D-37075 Goettingen Germany Phone: +49 551 397036 Fax: +49 551 397004 Email: mochs-at-gwdg.de
R.Rahbari, the last contact we had with Ultraclone was at the address you listed and their FAX is 44.1983.760263. This and 4700 other biotech companies contact info may be found at 'Company Index Search' at the website www.antibodies-probes.com. There are NO passwords necessary to use the address database. If you find any errors, please bring them to my attention via email. Thanks, Bob Weimer
RAHBARI, RAMIN wrote: } } Hello list members } } I wonder if anyone could help me get in contact with Ultraclone Limited. } They are the named producer of a particular antibody in an experiment I } am trying to duplicate. } } Ultraclone Limited } Rossiters Farm House } Wellow, Isle of Wight } PO 401-OTE, UK } } I have contacted them by snail-mail and had no response. Can anyone } provide an e-mail address or telephone or fax number? } } Any help or information would be greatly appreciated. } } Ramin Rahbari } Tel. 313-998-3383
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I have used both Umax and Microtek and have been happy with each. For evaluation reports you might try: http:\\www.zdnet.com This is the entry point. Couldn't get the exact (further into site) address while at work... Think java was crashing the NS 2.0 we run at work :(
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
Home woody.white-at-worldnet.att.net http://www.geocities.com/capecanaveral/3722
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I am in the market for a flatbed scanner and would like to here your opinion regarding choices of flatbed scanners (such as specific brand, model, cost,...etc). The scanner will be used for digitizing the EM micrographs or images which are to be either archived or used for quantitative image analysis.
I know that this is an old subject for this group, can someone help me find a summary list of the scanners that were discussed previously by this group? Lastly, Is there any new and better model available since the subject was last discussed? Your information will be greatly appreciated.
Hi, I work at Brigham & Women's Hospital, E.M. Lab in Pathology and Steve Mello from Histology gave me your e-mail about your problem with the thermal unit on your Lynx. I had the same problem several months ago and called for service because I have a service contract, but before the srevice tech. came we discovered the problem. There is a contact on the back of the unit for the thermal unit that became loose. It did not fall off so the alarm did not sound but it gave an incorrect reading to the unit and I cooked a few biopsies because of it. My telephone # is 1-617-732-7527 if you want to ask me anything about what happened. Christine Ridolfi
Serial transfer using Kermit is one way but there is another. The alternate is costlier, but faster - the "sneaker net". A similar (or same if you want to swap a lot) Syquest drive can be installed on a PC platform and special Kevex software used to cross copy files between the DEC format disk and the PC world. Such an installation requires a SCSI card in your PC and the Kevex programs RTCOPY and RTDIR (ectory). I am using the Adaptec 1542 SCSI adapter. Others may work, but have no experience or data. The programs run in DOS (or in W95 DOS window) and work much like the DOS commands "copy" and "dir". Syntex example from C:} ....
rtcopy dl2:name.typ c:\whatever\name.typ
A batch file to handle multiple files wouyld be handy, but I never conjured up one... Wild cards (*) are implemented in a limihed fasion... The pgm will interrogate you on each file to copy, like it or not.
Let me know if you have any other questions.... Woody
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
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We finally (thank god!) replaced our 10MByte Bernoulli disk drives on our Kevex 8000 system with SyQuest drives. Kevex supplied us with a program that converts spectra and images to .tiff files. Has anyone had experience with sending these files to a Windows-based pc? How is it done? Do I need Kermit or is there a simpler way?
South Bay Technology manufactures a research level parallel plate oxide etcher and complete data on the system will be mailed this afternoon for your review. Our unit is very comparable to the system you mentioned.
The South Bay Technology system, model number PE-150, is designed with a 150 watt, 13.56 MHz RF power supply and a six inch diameter electrode. U= p to a full six inch diameter sample can be used in this instrument. Two independent interlocked gas controls are included as well as digital displays for vacuum, forward power, reflected power, DC bias and time. Termination of the plasma discharge is by time and the unit includes a front panel set timer. All stainless steel construction is used and comm= on gases such as CF4, C3F6, Ar, and other chlorine or flourine based chemistries can be utilized. The system is table top mounted and include= s a two stage direct drive corrosive series rotary vane pump. Base pressur= e is typically 15 millitorr and operating pressure ranges from 60 millitorr=
to over 200 millitorr depending on the application.
Please feel free to call Steve Collins in our East Coast offices at 703-486-7999 or email at scollins-at-southbaytech.com . You can also reach = me in California at the numbers below. Information will be mailed including=
etch rates, uniformity of etch and general product design data. =
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by INTERNET:WINDLAND-at-odin.ssec.honeywell.com } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
We have a benchtop March Instruments parallel plate etcher. We are currently using it to etch off layers of dielectric between metal layers on our semiconductors. We are interested to find out if anyone has an etcher fo= r a similar purpose and what instrument they are using. We have not been abl= e to find any small etchers other than the March Inst. model. Thanks for you= r input. Mark Windland Analytical Services Solid State Electronics Center Honeywell Minneapolis, Minnesota 612-954-2845 fax 612-954-2040 e-mail: windland-at-ssec.honeywell.com {
I have references for cryo EM using the Tokuyasu method, but my question is at what step of fixation / cryoprotectant can you store tissues, and for how long? I realize there will be variations with tissue and antigen, and whether you are using glut., but what about straight Para-form? The other question is cryo EM on cell culture. I typically pellet into agarose rather than trying to get the cells to hold themselves together. Has anyone done cryo with agarose? Thanks.
Question: I am planning to perform EBSP on mullite fibers mounted in cold-set. What is a suitable etchant? I have been recommended HF but what concentration and etching time is required? Are there any alternatives to HF? Thanks
Question: Hi- I came across this and am wondering if you could reccomend 1) a good way to stain fungal tissue for visualization with confocal laser scanning microscopy 2) a good manual which deals with various fixation/ staining techniques for CSLM & epifluorescence. I'll bet this is interesting work for y'all.
Some 5 years ago I read a review article on high resolution phase contrast microscopy I think down towards 10nm resolution, unbelievable as that may seem. I think that the article was from Eastern Europe.
Is there any one out there currently working in high resolution phase contrast microscopy? We may have some funding in this area looking for a researcher. We are in New Zealand if that is any incentive! I look forward to hearing from you.
Dear Sir/Madam, This mail is to inform you that I wish to unsubscribe to the Microscopy Society. My need was temporary and I have obtained all the information I need. My mail account is also running out of space as I do not have access to a lot of space. Kindly ensure that I do not get any mails from now on. Thank you very much for all the info provided so far. Please terminate my account now. Thank you for your time Sincerely, Kiranam.C {kiranam-at-biosys.net; kiranam-at-mailexcite.com}
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Reply to: RE} Cryo EM on agarose pellets
Dear Ricky, It is safe to store the tissues in formaldehyde for extended periods of = time. If they are fixed in glutaraldehyde, you can safely store them in = buffer. We have stored the pieces of tissue in sucrose cryoprotectant = for about a week without problems, but be sure to mix the sucrose = solution while infiltrating to avoid sucrose gradients from forming in a = stationary tube. For the cell pellets, we generally use 10% gelatin to = hold them together and then treat the pellet the same as a tissue sample = (cut into pieces, infiltrate and freeze). Linda Chicoine Center for Cell Imaging Yale Univ. Dept. of Cell Biology New Haven, CT 06520-8002 203-785-3646 phone 203-785-7226 fax --------------------------------------
I have references for cryo EM using the Tokuyasu method, but my question is at what step of fixation / cryoprotectant can you store tissues, and for how long? I realize there will be variations with = tissue and antigen, and whether you are using glut., but what about straight Para-form? The other question is cryo EM on cell culture. I typically pellet into agarose rather than trying to get the cells to hold = themselves together. Has anyone done cryo with agarose? Thanks.
Rick L. Vaughn
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Greetings, When I switch from observing a secondary image to a backscattered one, in what direction to I go to get the best image? Is the accelerating voltage, working distance, tilt, spot size, final aperture and so on increased or decreased? If this depends on the sample, still is there some general directions to take when switching from a secondary analysis to a backscattered one?
I need advice on how to determine contents of 1-10 micron sized pores formed in steel.
The pores appear only after a particular sequence of heat treatments and are always associated with corners of cuboidal titanium nitride precipitates. We believe that for certain duration of time during the heat treatment the steel is under tension in the regions of pore formation.
We have speculated that (1) the pores may contain vacuum and nucleate and grow by coalescence of vacancies in regions of tensile stress concentration, (2) are filled with hydrogen coming out of solution in steel, (3) are filled with nitrogen from decomposition of the titanium nitrides (my sense is that thermodynamically this is unlikely). Any ideas on how to differentiate among these scenarios and/or can you suggest other possibilities?
The only method that I could come up with is by Auger spectroscopy on gas molecules adsorbed on surfaces of pores in samples broken in situ under vacuum, but I have no experience with high vacuum SEMs or with Auger. Would it work? Is any one aware of equipment capable of this in NE or Mid Atlantic USA?
I will be grateful for any thoughts, suggestions and leads.
Valdemar-at-fast.net
Valdemar Furdanowicz Homer Research Labs G165 Bethlehem Steel Co Bethlehem PA 18016.
Hmmm, best image? That would depend on your definition of best. I will define "best" as good signal-to-noise at lower mags.
Signal is MUCH improved with reduced working distance (~1/r**2). This assumes that your detector is mounted under the pole piece (as many are) and that you are not simply running your E-T detector in BSE mode. There should be no resolution sacrifice.
Signal will go up with beam current (~I) (spot size, final aperture) with some loss in spatial resolution.
The signal generally goes up with increased accelerating voltage but spatial resolution may decrease due to a larger scattering volume.
Generally, we have to open the beam up quite a bit to get enough BSE signal. We measure a current of ~0.3 nA at 20 kV and 25 or 39 mm WD. I would like to reduce working distance, but we are often doing EDS simultaneously and thus it is fixed by our detector.
Hope this helps.
At 08:45 AM 1/22/98 -0500, you wrote: } Greetings, } When I switch from observing a secondary image to a backscattered one, } in what direction to I go to get the best image? Is the accelerating } voltage, working distance, tilt, spot size, final aperture and so on } increased or decreased? If this depends on the sample, still is there some } general directions to take when switching from a secondary analysis to a } backscattered one? } } Thanks, Mark Darus ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
Does anyone have experience sectioning tissue on Aclar film? I getting ready to embed some pollen that has been stuck to Aclar film. I want to know if Aclar is easy to embed and section. I'm use to sectioning tissue grown on dialysis tubing. Your comments are greatly appreciated.
best regards, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I'm going to be involved in TEM characterisation of coatings for working on the ultraviolet range. The materials extensively used as UV-Coatings are fluoride compounds as Mg2F, AlF3, LaF3 and related...I know that these materials should be hygroscopic and instable.
Any help on material preparation for TEM examination will be extremely appreciated.
Thank you very much in advance
F. Peir=F3 *******************************+ Francesca Peiro
EME, Enginyeria i Materials Electronics Dpt. Fisica Aplicada i Electronica Universitat de Barcelona Avda. Diagonal 645-647 08028 Barcelona, Spain
In response to request for fungal stain. One should be able to use a Gomori Methenamine Silver stain(called GMS stain). The silver helps visual fungal hyphae and should provide differential contrast both photons (light micros.) and electrons (E-M). Worst case scenario you might have to remove the plastic if embedded in epon-araldite (for example) prior to staining. Also look under methenamine silver staining for EM as this has been used for glycosaminoglycans and should stain fungi nicely too.
A complete answer would make a book.... Depends on the examination goals (penetration, Z-range etc.), the specimen, detector type, etc. Generally BSE will require more beam current than SE. An untilted (normal to beam) specimen surface will typically maximize collection of BSEs. I could go on, but....
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
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Greetings, When I switch from observing a secondary image to a backscattered one, in what direction to I go to get the best image? Is the accelerating voltage, working distance, tilt, spot size, final aperture and so on increased or decreased? If this depends on the sample, still is there some general directions to take when switching from a secondary analysis to a backscattered one?
} -----Original Message----- } From: bent-at-unixg.ubc.ca [SMTP:bent-at-unixg.ubc.ca] } Sent: Wednesday, January 21, 1998 5:44 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: staining fungal tissue? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Email: bent-at-unixg.ubc.ca } Name: Elizabeth Bent } } Question: Hi- I came across this and am wondering if you could } reccomend 1) a good way to stain fungal tissue for } visualization with confocal laser scanning microscopy } 2) a good manual which deals with various fixation/ } staining techniques for CSLM & epifluorescence. I'll } bet this is interesting work for y'all. } } } ---------------------------------------------------------------------- } ----- }
---------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of=20 America To Subscribe/Unsubscribe -- Send Email to=20 ListServer-at-MSA=2EMicroscopy=2ECom=20 ---------------------------------------------------------------------- =20 Mark Darus wrote: =20 Greetings, When I switch from observing a secondary image to a backscattered one, in what direction to I go to get the best image? Is the accelerating=20 voltage, working distance, tilt, spot size, final aperture and so on=20 increased or decreased? If this depends on the sample, still is there some= =20 general directions to take when switching from a secondary analysis to a=20 backscattered one? =20 Thanks, Mark Darus =20 =20 BSE imaging is usually employed to observe differences in atomic=20 number contrast=2E Therefore the conditions to excite more BSE's can=20= be=20 used=2E Working distance and probe current are probably the most=20 important=2E The shorter working distance is important because BSE's=20 can be considered a line of sight signal and the closer you get the=20 source of BSE's to the detector the stronger is the signal=2E Unlike=20 the E-T detector which has a voltage applied to attract SE's, the BSE=20 detector, if your using a solid state type does not attract BSE's by=20 using a bias voltage, but by proximity to the sample=2E Increasing th= e=20 probe current by changing the spot size or increasing the primary beam= =20 electrons getting to the sample by using a larger aperture will also=20 improve the BSE signal=2E If you also need good resolution with BSE=20 then some compromises will need to be made=2E =20 =20 Since the BSE's come from an area of the sample where the beam is=20 beginning to spread the BSE resolution will not be as good as SE=20 imaging in most cases but if you are using lo kv where spreading is=20 less the BSE and SE resolution will be similar=2E =20 If you are doing EDS along with BSE and SE imaging you will most=20 likely be using 15-25 keV to excite x-rays=2E In this situation, the=20 easiest way to get a good BSE image to correlate with the SE image is=20 just to use a larger apreture without changing anything else=2E =20 =20 Hope this helps =20 John Humenansky Braun Intertec 6875 Washington Ave=2E So=2E Minneapolis, MN 55439
I want to thank everyone who responded to my request for techniques to make swollen gel particles visible. Very helpful information - I will make use of it.
A couple of responders volunteered to do a few tests on samples. I will contact you about this soon.
Cut some small notched samples, have a cooling device arrangement for them to get them under the ductile-brittle transition temperature in a vacuum, and break them with an impact hammer in a vacuum chamber that has an residual gas analyzer. You should be able to see what gas is trapped in the pores by the increase in the peak pressures. However, the diffusion of hydrogen in steels is very high and you probably will not see an increase in the hydrogen from this.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I need advice on how to determine contents of 1-10 micron sized pores formed in steel.
The pores appear only after a particular sequence of heat treatments and are always associated with corners of cuboidal titanium nitride precipitates. We believe that for certain duration of time during the heat treatment the steel is under tension in the regions of pore formation.
We have speculated that (1) the pores may contain vacuum and nucleate and grow by coalescence of vacancies in regions of tensile stress concentration, (2) are filled with hydrogen coming out of solution in steel, (3) are filled with nitrogen from decomposition of the titanium nitrides
(my sense is that thermodynamically this is unlikely). Any ideas on how to differentiate among these scenarios and/or can you suggest other possibilities?
The only method that I could come up with is by Auger spectroscopy on gas molecules adsorbed on surfaces of pores in samples broken in situ under vacuum, but I have no experience with high vacuum SEMs or with Auger. Would it work? Is any one aware of equipment capable of this in NE or Mid Atlantic USA?
I will be grateful for any thoughts, suggestions and leads.
Valdemar-at-fast.net
Valdemar Furdanowicz Homer Research Labs G165 Bethlehem Steel Co Bethlehem PA 18016.
A student in our EM lab would like to try out a protocol which involves use of hydrofluoric acid. The protocol calls for single cells, attached to a glass cover slip, to be frozen, freeze dried, and rotary shadowed. The replica is then released by immersion of the cover slip in hydrofluoric acid. I hear that HF is nasty stuff, and would like to hear from anyone who has experience with 1) this particular application (concentrations, times, rinsing) and 2) safety issues relating to the use of HF in general. What precautions should we take, how worried should we be, etc.
Thanks for your help.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
There are many reasons why one would want to switch from SE to BSE observation. As it was already mentioned, this topic can easily fill a book.
I just wanted to point out one more aspect: on some SEM's there is the possibility to mix SE and BSE signals with variable ratio, thus increasin= g the perceived image quality. The advantages are (to name just a few): lower impact of those high-contrast areas that frequently appear in the SE-only images; lower signal from the sample's background (lines and cracks on the adhesive (carbon) tape; the "flat" BSE image gets some topographic contrast from t= he SE portion; etc.
This works especially good with things like diatomes, radiolarians etc., which are difficult to coat completely with carbon or metal, and therefor= e can produce ugly highlights when observed with SE alone.
Quidado. You are correct, Mg2F, AlF3, and LaF3 are unstable, susceptible to beam damage, and difficult to work with. They easily dissociate = into metal rich phases and evolve fluorine. I've found a number of = published images of MgF2 that are primarily artifacts of sample preparation and beam damage. =20
Due to the chemical nature of the instability, I've chosen to prepare TEM cross-sections of UV coatings (Mg2F, AlF3, and LaF3) using ultramicrotomy with a diamond knife. There are two things that are essential for sectioning hard materials (including thin films, small particles and crystals): 1) Minimize the amount of material to be cut. Prepare samples with a minimum amount of substrate, and orient them in the embedding media for a cross-sectional view. Cut the blockface on a microtome taking small cuts to reduce stress to the interface between epoxy and sample. End up with a blockface of less than 100 um across (50 um preferred!). 2) Improve the adhesion of the embedding media to the sample. Many embedded hard materials fail by pulling out when sectioned due to poor adhesion. First, I have found that Spurrs works best but there = are other compounds that may work. Second, it's essential to improve the adhesion of the epoxy using an adhesion promoter such as Z-6040 from = Dow Corning (a silane copolymer).
To minimize artifacts these unstable fluorides require careful TEM techniques working under minimum dose conditions. As a reference, you should always first take very low magnification images of your sections and work up to higher magnifications. Don't focus at higher magnifications to improve lower mag images.
good luck, Phil Swab Advanced Coatings Division, ART inc.
} ---------- } From: paqui-at-iris1.fae.ub.es[SMTP:paqui-at-iris1.fae.ub.es] } Sent: Wednesday, January 21, 1998 11:14 PM } To: Microscopy-at-Sparc5.Microscopy.Com; } Microscopy.com-at-Sparc5.Microscopy.Com } Subject: TEM of fluorides } =20 } = ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America=20 } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } = ---------------------------------------------------------------------- } -. } =20 } Dear Colleagues, } =20 } I'm going to be involved in TEM characterisation of=20 } coatings for working on the ultraviolet range. The materials=20 } extensively used as UV-Coatings are fluoride compounds as Mg2F,=20 } AlF3, LaF3 and related...I know that these materials should be=20 } hygroscopic and instable. } =20 } Any help on material preparation for TEM examination will be=20 } extremely appreciated. } =20 } Thank you very much in advance } =20 } F. Peir=F3 =20 } *******************************+ } Francesca Peiro } =20 } EME, Enginyeria i Materials Electronics } Dpt. Fisica Aplicada i Electronica=20 } Universitat de Barcelona } Avda. Diagonal 645-647 } 08028 Barcelona, Spain } =20 } Tel. (34-3) 402 11 39 } Fax. (34 3) 402 11 48 } e-mail: paqui-at-iris1.fae.ub.es } **************************** } =20
Dear Marie, } } A student in our EM lab would like to try out a protocol which involves use } of hydrofluoric acid. [snip] } and 2) safety issues relating to the use of HF in general. } What precautions should we take, how worried should we be, etc. } HF is, indeed, nasty. Obviously do everything in a hood. HF does not eat polyethylene, so use poly gloves. That said, HF is just a mineral acid, so there is no need to panic. The fumes are bad, but so are those from HCl, HNO3, etc. If the protocol calls for dilute HF (~1N), there is less concern than if it calls for concentrated HF. Once the HF has been diluted from stock, the gloves and a well-ventil- lated hood should be sufficient precautions. Good luck. Yours, Bill Tivol
Yes, Aclar film is relatively easy to section. I've cut lots of cells and tissues which have been on the stuff. What I find helpful is as follows:
1. Flat-embed the specimen and polymerize as usual (assuming you want to cut the Aclar in cross-section.
2. Orient and trim, preferably using precision trimming on the microtome with glass knives or an old diamond knife, so that all sides are clean and "polished".
3. Cut sections the long way. Trim the block so the first and last parts to contact the knife are the ends of the Aclar, then make sloping cuts at the top and the bottom of the block so the face gradually makes a transition from resin+specimen+Aclar to just Aclar by itself. This means the block face will be rather tall and narrow...just the opposite of the way you trim most blocks!
3. Then, come in from the underside of the Aclar, trimming away about half of its thickness. The idea here is to make the top, bottom and one side of the block face pure Aclar, and the reason to do so is to minimize curling of the sections and splitting of the Aclar away from the main part of the section. Below is my feeble attempt to "draw" the block face:
_ /| | / | |------Aclar, with specimens surrounded by resin / | | (this "bottom" side is trimmed to half-thickness) | *| | | *| | | *| | | *| | | | | \ | | \ | | \|_| | | Sectioning direction V ________________________ Knife
If your e-mail reader rearranges all of my fancy artwork, please contact me off-list. I can fax you the drawing.
Best of luck! Bob ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Cryostats / Microtomes / Tissue Processors / Embedding Centers / Slide Stainers / Glass Coverslippers / Microscopes (Representing Leica since 1967) / Fiber Optic Systems / Linear Measuring / Micromanipulation (Linear Encoded, Video) / Image Analysis, Archiving, Capture / Video & Video Printers (Cooled CCD, Digital, RGB, Super VHS, 3-chip) / Vibration Isolation Systems / Programmable Stages / Heating & Cooling Stages
I am new to the field and am wondering if there is any software for X-ray deconvolutions for EDXA-SEM system? IF it has to be done manually, could you give any refences on how to do it best?
I also would like to get in touch with people who study particles on vegetative surfaces to learn and share some thoughts.
FEI Company in Hillsboro, Oregon has the following position available:
FIB / SEM Applications Engineer
Job Summary: Develop and demonstrate to various customers the full capabilities of FEI FIB and SEM systems and related equipment
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* BS/equivalent in Physics, Material Science or related discipline * 2 years experience operating SEM, TEM, EDS * Exceptional interpersonal communication skills * Computer literate, experience with MS Windows programs including but not limited to MS Word, Excel, and Visual Basic * Willing to work in clean rooms * Eligible to work in the United States * Eligible for passport and willing to travel up to 25%, domestic and overseas * FEI Company is a world leader in providing superior field emission products and applications to our customers throughout the world. We build world-class electron microscopes and related products using ion and electron beam technologies. Our products are used by the semiconductor and material science industries as well as various medical and scientific institutions. We're located in the midst of the Silicon Forest, near the intersection of Highway 26 and Cornelius Pass. FEI is headquartered in Hillsboro, OR with nearly 1,000 worldwide employees.
Employees at FEI work in an open, team oriented environment and have the opportunity to apply their skills to a wide variety of challenging problems in manufacturing, engineering and business. Work alongside some of the world's foremost experts in the area of field emission technology!
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Please submit resume to Lisa Olivia either by FAX: 503.640.7509 or email: lolivia-at-feico.com.
Lisa Olivia Recruiter FEI Company PH) 503.844.2601 FAX) 503.640.7509 email: lolivia-at-feico.com
} Greetings, } When I switch from observing a secondary image to a } backscattered one, } in what direction to I go to get the best image? Is the } accelerating voltage, working distance, tilt, spot size, final } aperture and so on increased or decreased? If this depends on the } sample, still is there some general directions to take when } switching from a secondary analysis to a backscattered one? } } Thanks, Mark Darus }
You'll need to be more specific regarding the materials and goals of your work if you want more specific answers. Basically, by it's nature, BSE is a three dimensional process with interactions taking place beneath the surface of your sample. Like x-ray production in a sample, the BSE electrons can come from a volume within the sample much larger than the exciting electron diameter. This effect will be enhanced by higher accelerating voltages.
For best spatial resolution you might want to experiment with higher sample tilt. The simplest explaination here is that at high tilt angles, BSE electrons scattered at greater depths in the sample will have to come through increasingly larger amounts of sample material to reach the detector, leaving the majority of detected electrons those emitted from near the surface.
This brings us to a second concern - the detection efficiency. Backscatter detectors vary greatly in their collection efficiencies. Older, or less expensive, detectors are often much less efficient at collecting BSE and produce a much grainier image than secondary detectors at a given set of operating conditions. Decreasing the condensor currents can produce a higher beam current, and thus higher electron emission and 'smoother' looking images, but at the expense of spatial resolution. Ditto increasing final apeture size. BSE collection will be adversely affected by higher tilt angles.
Remember that the slow scan speeds used taking a micrograph create a much smoother photographic image than the visual rate image you use when setting up for taking the photo. Generally, when looking for the best resolution in any micrograph you'll want to work with a visual rate image that is very grainy, yet just good enough to accurately focus and stigmate on. Using slower visual rates can help here.
Finally, remember the signal contrast mechanisms of BSE. Average atomic number spatial differences provide the primary contrast, while surface topology is a secondary contributor. BSE yield is higher as the average atomic number increases. Contrast in a BSE image increases as localized differences in average atomic number increase. Don't expect to be able to produce much of an image if you are looking at purely biological materials. While antigen-labeled gold microspheres will show up very well against tissue, you won't see much detail in the tissue itself. Metallic grains can show very well, but you will have to use a high electronic amplification (gain) to bring out the contrast in many structures.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
RMC manufactures new Universal Specimen Holders that will fit your MT-2B (also all subsequent models by DuPont and RMC as well as Ultracuts with = a $28 adapter). This holder will accept flat blocks and both sizes of Beem blocks.
The part number is 70930, cost is $215, they are in stock. =
There is a second version that has a conical mirror behind the block and = a port for a light pipe to provide transillumination. You obviously need a light source to make this work.
Steve Miller RMC Phone: 520-903-9366 Email: Steve.Miller-at-RMC-Scientific.com Website: http://www.RMC-Scientific.com/microtomes/
TO: David Johnson FROM: Steve Miller RE: Bellows repair
If you are not successful in locating a complete assembly it is possible = to repair the bellows yourself. It requires purchasing the bellows material and using a hot plate and holding jig but nothing exotic. =
See if you can remove the defective bellows from the machined end pieces = by heating to melt the solder.
I am planning to order a high-enegy mill to prepare powders with size down to at least 0.05 microns. If you happen to know this kind of machinces and the producers, please tell me a little detailed imformation on the model, price, company and company phone number.
Microscopists, I am being asked to fix and section fish otoliths for TEM. Does anyone have a protocol? Or opinions? For instance do I have to decalcify? If so is formic acid better than EDTA? How do I determine the end point of decalcification. Can I use Epon or would another embedding media work better? How much damage can I expect this to do to my diamond knife? I did get a few tips from the University of Florida's web site but would value input from anyone experienced in working with bone. Thanks very much! Kim
-------------------
Kim DeRuyter Histology and Electron Microscopy PO Box 755780 University of Alaska Fairbanks, Alaska 99775-5780 fnksd1-at-aurora.alaska.edu (907)-474-5452
I am afraid that the hazards of HF were understated. A decent spill on your body can be lethal. The "appropriate" labwear apparently is a full body apron, gloves and a protective visor -- remember the potential legal issues. You also need a neutralizer (which costs a bit).
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
I am looking for an older Philips SEM in any condition. The vintage I am interested in includes the models 515, 525 and 535. Please contact me by e-mail or phone if you have any leads. Thanks, Al Sicignano, mks-at-cyburban.com, phone/fax 914 241 0864.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hey, y'all - } } Does anyone have experience sectioning tissue on Aclar film? I getting } ready to embed some pollen that has been stuck to Aclar film. I want to } know if Aclar is easy to embed and section. I'm use to sectioning tissue } grown on dialysis tubing.
Hi Beth, I have used Aclar many times. It is perfect! It embeds beautifully. It reacts with nothing. It sections beautifully. It easily peels off the polymerized epon if you want. I love it. Sally Shrom
} Your comments are greatly appreciated. } } best regards, } beth } } ************************************** } Beth Richardson } EM Lab Coordinator } Botany Department } University of Georgia } Athens, GA 30602 } } Phone - (706) 542-1790 } FAX - (706) 542-1805 } Email - beth-at-dogwood.botany.uga.edu } ************************************** } } }
Do any of you have a contact telephone number for distributors of Cargille's Immersion oil. A number in the UK/Europe would be ideal. I have tried Nikon U.K., but they only stock type DF oil and I am looking for the non-autofluorescing type.
Many thanks
Dr. Giles Sanders Laboratory of Biophysics and Surface Analysis School of Pharmacy University of Nottingham
After reading Laurie Marks' post I went to the safety office and looked up HF in the CRC Handbook of Laboratory Safety. Laurie was correct about the hazards of HF, and I understated them. The Handbook says, "It is an extremely dangerous material and all forms, including vapors and solu- tions, can cause severe, slow-healing, burns to tissue. At concentrations of less than 50%, the burns may not be felt immediately, and at 20% the effects may not be noticed for several hours." Also, "Fluoride ions readily penetrate the skin and tissue and, in extreme cases, may result in necrosis of the subcutaneous tissue which eventually may become gangrenous. If the penetration is sufficiently deep, decalcification of the bones may result." It goes on to say, "Dilute solutions and vapors may be absorbed by clothing and held in contact with the skin, which will probably not result in an immediate sensation of pain as a warning but eventually may lead to skin ulcers which, again, may take some time to heal. A generalization might be made here about absorbant clothing. In many instances, as in this case, absorbant clothing which can retain toxic materials and maintain them in close contact with the skin may be worse than no protection at all, changing the exposure from a transient phenomena (sic) to a persistant one." I'm sorry about my earlier post. Yours, Bill Tivol
I have been working on ultrastructure and micro-chemistry of fish otolith for many tears in Hawaii. I have developed a protocol especially for ultramicrotoming of undecalcified otolith. The following is my method:
1) Double embedding: Pick up a small piece of otolith ( {0.5mm), embede it with Epon resin. Grind (#600 through #1200 sand paper). As soon as the grinding plane reaches the otolith, stop. Put a drop of Spurr resin on top and cure it. Then go grind-embed back and forth 3-5 times. By doing this, the resin around otolith is compressed and stronger support could be achieved.
2) Sectioning: I used Ultracut E for sectioning and it worked very well. Use very low cutting speed, low level of water surface, and diamond knife. Due to the high hardness of otolith, the otolith is acturally not sectioned, but micro-broken. The enhanced embedding procedure can prevent otolith from movement and turning, and hold the material of otolith on wherever it was in otolith. You can see crystal-composed image under TEM. The daily increment rings also can be seen in details under x10K. The sections are suitable for EDS and EELS analysis as well as element filter mapping.
3) Decalcification: Fish otolith is high calcified material with about 90% of inorganic. After decalcification, without support from inorganic crystalls, the organic residure will be in form of pieces or dissolved. It can not form a solid structure represent the structure of otolith. Therefore, young otolith from few-day's larvae may have more organic materials in otolith. After decalcification, you can see something left, but it is in the form of gel. It is impossible to go through fixation-dehydration and embedding procedure. I have never been succesfull for decalcified otolith. You can decalcify otolith on surface of sectioned block, then go through the fixation-dehydration and embedding procedure on top of block. But for otolith research, the information of micro-chemical components, location and distribution is much more important than ultrastructure. The number of grow rings and its structure can be better determined by light microscope with polarized ligh source and SEM with SEI or BEI image models.
Hope it helps. ****************************************** Zhiyu Wang Electron Microscope Lab and Imaging Center Western Kentucky University(WKU) Bowling Green KY 42101
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Microscopists, } I am being asked to fix and section fish otoliths for TEM. Does anyone } have a protocol? Or opinions? For instance do I have to decalcify? If so } is formic acid better than EDTA? How do I determine the end point of } decalcification. Can I use Epon or would another embedding media work } better? How much damage can I expect this to do to my diamond knife? I } did get a few tips from the University of Florida's web site but would } value input from anyone experienced in working with bone. Thanks very } much! Kim } } ------------------- } } Kim DeRuyter } Histology and Electron Microscopy } PO Box 755780 } University of Alaska } Fairbanks, Alaska 99775-5780 } fnksd1-at-aurora.alaska.edu } (907)-474-5452 } } }
Thank you for your correspondence regarding the Emitech cryo system. We do not have an Emitech cryo system at the University of South Africa in Durban. I believe you were operating a competitive system that is in excess of 11 years old that was interfaced to a JSM35. So the problems you encountered were not that of the Emitech system.
If you would like to have the opportunity to evaluate our system, we would be more than happy to accomodate you.
Regards,
John Fitzpatrick President & CEO Emitech U.S.A., Inc.
Dr. Sanders, try the following for Cargille RI Liquids:
Sara Mark McCrone Scientific, Ltd. McCrone House 155A Leighton Road London NW5 2RD UNITED KINGDOM
011 441 71 267-7199 voice 011 441 71 267-3383 fax
Dr. Giles Sanders wrote:
} Dear Microscopists, } } Do any of you have a contact telephone number for distributors of } Cargille's Immersion oil. A number in the UK/Europe would be ideal.
{snip}
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
The US office of Cargille's can probably help you. Their number is 201-239-6633. Jean Behlen is a good contact.
Get back to us if we can be of further assistance.
Regards,
Elinor Solit The Microscope Book 800-440-0311
On Fri, 23 Jan 1998, Giles Sanders wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear Microscopists, } } Do any of you have a contact telephone number for distributors of } Cargille's Immersion oil. A number in the UK/Europe would be ideal. } I have tried Nikon U.K., but they only stock type DF oil and I am } looking for the non-autofluorescing type. } } Many thanks } } Dr. Giles Sanders } Laboratory of Biophysics and Surface Analysis } School of Pharmacy } University of Nottingham }
See article by Bohne and Harding in Hearing Research 109(1997) 34 - 45. On page 39 she describes taking thin, Durcupan embedded segments of mouse cochlear duct and decalcifying them in EDTA(0.2M) THROUGH the polymerized plastic. These are were viewed with LM and TEM. Julie Gross Dept. of Anatomy University of CT Health Center Farmington, CT 06030 860-679-2463 jgross-at-neuron.uchc.edu
The EM Core lab at the Interdiciplinary Center For Biotechnology Research, University of Florida will be holding a 4 day Short course on Scanning Electron Microscopy for Biologists Mar 9-12 1998. For more information please check the following web site. Have a good day.
www.biotech.ufl.edu/~emcl/news/course.html
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
All- Remember that abstracts are due on FEBRUARY 1st (in nine days!) for the Microscopy and Microanalysis '98 meeting to be held in Atlanta GA from July 12-16th, 1998. Check out the website at :
for more details on planned symposia as well as abstract submission. Kathi
Kathleen B. Alexander Metals and Ceramics Division Oak Ridge National Laboratory P.O. Box 2008 MS-6376 Oak Ridge, TN 37831-6376 PH (423) 574-0631 FAX (423) 574-0641
To: Jerome Jasso Microscopy ListServer Mike Dauzvardis
Mike forwarded Jerome Jasso's note to me for comment regarding Formalin (HCOH) fixed gross anatomical specimens. The Anatomy Board administers the Body Donor program and prepares cadavers and specimens for medical education programs and research studies. Here at the Univ. of MD, Anatomical Division, I also perform special preparations of gross specimens including plastination.
In many research and tissue preparation labs there is still a need and use for the aldehydes, whether it be Formaldehyde in path, Gluteraldehyde for EM, or Paraformaldehyde for histology, etc. In "gross anatomy" study labs, there is the need to prevent the dessication of the already anatomically fixed or embalmed specimen while it is out for extended time for study use. We use a "wetting fluid" to keep the specimens moist, inhibiting dessication /dehydration and to alos prevent airborne or cross contamination.
The Maryland Anatomical Embalming Fluid is composed of the histoic chemical ingredients: Phenol (5.3%), Formaldehyde (1.8), Alcohol (10%) and Glycerine (10%) by volume. The Fluid is shipped as a concentrate (1 part MAEF/ 2 parts H2O. I don't pay to ship water! We order in 50 gallon drums but it can also be ordered in smaller quanites, 20 liter carboys, for example. The Board has not compounded its formula for many years. Instead we have a commercial chemical company (Hydro Chemical Co. -at- 800-345-8200 in Pennsylvania compound it for us. This is for both reasons of staff safety and it is more cost-effective. You can call them for information.
The "Maryland Wetting Solution" is the same formula as the Embalming Fluid, only there is "NO" Formaldehyde in the solution at all. Phenol is a far better agent that is cidal to all virus, bacteria, fungi, and molds at the percent used in the formula. Formaldhyde can not make the same claim...molds can still grow and the Creutzfeldt-Jakob (slow-virus) has been shown to be Formalin resistent. Phenol (liquid carbolic acid) as the Chlorosptic Throat Wash commercial states.....there's nothing better to kill the bugs! Check the ingredients in it .. I was surprized.
The wetting fluid is not used for BRAIN. We fix whole brains in a 20% HCOH Solution (Formalin) and after a thorough fixation (which can be speeded up with glacial acetic acid, i.e. Dietrich's Solution)). The "wet" brains can be stored in a 1-2% Formalin solution, or can be plastinated whoe, in part, or sliced to render dry, fumeless, durable, NON-TOXIC specimens that can be handled.
There are other non-formalin solutions available, many adveritse as "Formaldehyde free" , such as NO-TOX, many are alcohol based. I have found that some may work for histology (depending on the tissue) but most don't work for gross specimens. There are solutions now avialable that will "wash out" the HCOH in the tissue to reduce extended toxic exposure. INFUTRACE (tm) BY S&S Co. of Georgia, distributed by Action Products, Att: Mr. Gaylan Hayes -at- 800 862-5326.
I could go on but I hope this will answer some of the concerns.
Ronn Wade
Ronald S. Wade, Director (I.S.P. Treasurer) Anatomical Services Division / State Anatomy Board School of Medicine, University of Maryland 655 West Baltimore St. BRB B-024 Baltimore, Maryland 21201-1559 Phone: Voice: (410) 706-3313 (410) 547-1222 (Anatomy Board) Fax: (410) 706-8107 Home: (410) 987-1869
I'd like to extend my "THANK YOU" to all that replied to my request for film scanner recommendations. I received some very helpful information and have requested more information from specific vendors.
Again, THANK YOU very much.
Annette Andrews, EM Lab. PAI - NCTR, MC923 3900 NCTR Road Jefferson, AR 72079 e-mail: aandrews-at-nctr.fda.gov Phone: 870-543-7039
I am looking for a protocol for processing sea urchin sperm for TEM/SEM and immunogold label. I'm interested in actin/ microtubules in the tail region.
******************************************************** * Raj Patel * * Dept. of Pathology * * Robert Wood Johnson Medical School * * 675 Hoes Lane, Piscataway, NJ 08854 * * * * voice (732) 235-4648; Fax -4825 * * E-Mail rpatel-at-umdnj.du * ********************************************************
We are trying to accomplish In situ hybridization on LRWhite thick sections on gelatin coated slides (slides are immersed in 1% gelatin sol'n and baked at 60C overnight) but they always float off. We cannot heat the sections; any suggestions?
David O'Neil tel: (902) 426-8258 National Research Council of Canada fax: (902) 426-9413 Institute for Marine Biosciences 1411 Oxford St. Halifax, Nova Scotia B3H 3Z1 Canada email: david.o'neil-at-nrc.ca
Is any one putting to pasture their Bernoulli 44 or 90 or 150Mb removable media hard drives? Is there a chance you are willing to part with any of the removable cartridges? Or do you know anyone who is?
Thanks in advance for any direct responses. Ron Oleka roleka-at-octonline.com Tel 905-297-2222 ext2304
Kim - For the uninitiated, otolith is a bone in fish's ear, now used to estimate the age of fish. Otolith have growth rings somewhat like trees). I doubt that there will be anything left after decalcification. Try it. I suggest that you use say 10% ascorbic acid in buffer, it's milder but EDTA certainly will do. A diamond knife will cut undecalcified otolith better than bone, but use a "hard" mixture of whatever your favourite embedding medium. Block size should be very small, but it would be anyway unless you are working on whales.
Microprobe analysis suggests that the rings are visible because of differing crystalline conditions as no elemental variations is detectable across the otolith. Since spatial resolution of X-rays is much better in TEM, you may be able find elemental variation and certainly you should obtain greater morphological details then is possible in SEM. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
-------------------------- } Microscopists, } I am being asked to fix and section fish otoliths for TEM. Does anyone } have a protocol? Or opinions? For instance do I have to decalcify? If so } is formic acid better than EDTA? How do I determine the end point of } decalcification. Can I use Epon or would another embedding media work } better? How much damage can I expect this to do to my diamond knife? I } did get a few tips from the University of Florida's web site but would } value input from anyone experienced in working with bone. Thanks very } much! Kim
} Kim DeRuyter } Histology and Electron Microscopy } PO Box 755780 } University of Alaska } Fairbanks, Alaska 99775-5780 } fnksd1-at-aurora.alaska.edu } (907)-474-5452 } }
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Mark Krass wrote: ===================================================== I have a question regarding the sectioning of brittle polymers. I am interested in observing the interior sectioned structure of a particular polymer and examine by SEM. I don't believe this material can be embedded in epoxy due to its porous and brittle nature. Any suggestions would help along with a technique to observe the interior morphology. ===================================================== Whether you can or can not embed the polymer depends mainly on the nature of the porosity, that is, is it open celled or closed cell. It if it open celled, you do have a good chance of success, if closed, then less of a chance to zero chance. Second concern is to make sure your embedding system is not in any way swelling or other wise interacting with the polymer system under study. If that is happening, at least it has been our experience, it is difficult to detect by SEM. That is why you should save the sections that come off for possible examination by TEM at some later time. Such interactions, if occurring, are much more readily seen by TEM than SEM. Lastly, if you do the embedding under vacuum, you should in most cases be able to get good infiltration and a good "block". We ourselves use our own SPI-Pon 812 brand of resin but most of the so-called "Epon substitute" resins would probably work in an equivalent way. We like this system for this particular application because there seems to be less interaction with the polymer being embedded.
Finally, you will increase your chances still further by using a diamond knife as opposed to glass knives.
Disclaimer: Structure Probe, Inc. performs these kinds of studies for clients as a laboratory service.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} After reading Laurie Marks' post I went to the safety office and } looked up HF in the CRC Handbook of Laboratory Safety. Laurie was correct } about the hazards of HF, and I understated them. The Handbook says, "It is } an extremely dangerous material and all forms, including vapors and solu-
snips ...
} made here about absorbant clothing. In many instances, as in this case, } absorbant clothing which can retain toxic materials and maintain them in } close contact with the skin may be worse than no protection at all, changing } the exposure from a transient phenomena (sic) to a persistant one." I'm } sorry about my earlier post. } Yours, } Bill Tivol
I read up a safety book on HF years ago. The details were bad enough but what really turned my stomach was the recommended treatment - immediate injection into the centre of burns or suspected burns of calcium gluconate (??).
I've never gone anywhere near the stuff since then - I sooner work with high activity radiated material, or cyanide.
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
We are seeking a Senior Circuit Design Engineer for the design of coating measurement instruments. The technologies involved are x-ray fluorescence, eddy current, Beta backscatter, and ultrasound.
Some experience with x-ray detector electronics, preamplifiers, and MCA electronics is desired. Experience with eddy current and ultrasonic circuitry is a plus.
Our company, CMI International, is one of the largest manufacturers of coating measurement equipment. Our growth rate has been double digits for over 10 years. Our location is Elk Grove Village, IL. See our Web site at cmiinternational.com
Please send resume to rklein-at-cmiinternational.com or fax to Human Resources Manager at 847-439-4425.
I'm doing my thesis at the Aachen University of Applied Sciences. My research involves the actin-network in human blood cells. I've been desperately seeking for a method to stain the actin network of lymphocytes with a fluorescence in LIVING cells, but i can't find a method.
Does anyone know about a method of staining the actin-network of living cells for fluorescence mikroskopy ?
Dear Microscopists, Is there a method which would allow to measure, or preferably, image in situ sodium concentrations in ACIDIC compartments of a cell? The expected concentrations are in the millimolar range.
Meeting title: "Scanning Probe Microscopy in Biomaterials Science, Dentistry and Medicine"
Aim: The aim of this meeting is to give an overview of the use of scanning probe microscopy (SPM) in biomaterials science, biotechnology, dentistry and medicine. Beginners in the field of SPM and experts will be brought together. The meeting will be rounded off with practical demonstrations.
Meeting Format: A one day meeting with invited presentations, posters and practical demonstrations.
Date 2 April 1998
Venue Redwood Lodge Hotel and Country Club, Bristol, UK.
Fees There will be a registration fee of 10 GBP per delegate to be paid on arrival at the meeting. Digital Instruments (UK) Ltd. have kindly agreed to sponsor the rest of the costs of the meeting.
Expected Audience: The meeting is aimed at a distinguished academic and industrial audience with an interest in biomaterials science, biotechnology, dentistry and medicine.
Speakers: High calibre international and national speakers. A list of speakers is available under http://www.dent.bris.ac.uk/research/materials/Speakers.html
Organisers Dr. rer. nat. Klaus D. Jandt, Mr. Richard W. Vowles (University of Bristol); Digital Instruments (UK) Ltd.
More Information/ Registration/ Programme: Can be found under http://www.dent.bris.ac.uk/spm.htm
I have a catalog of "proteins and reagents for investigating the cytoskeleton". In France the products are sold by TEBU but you can find all the information you need : by phone : cytoskeleton , Denver USA : (303)322 2254 by e-mail : technical service : tservice-at-cytoskeleton.com customer service : cservice-at-cytoskeleton.com
I never tested any of these products up to now so let me know if some products work very well Have a nice day, -- =
************************************************************** * Dani=E8le SPEHNER, CJF 94-03 INSERM * * Etablissement de Transfusion Sanguine de STRASBOURG * * 10 rue Spielmann, B.P. 36- 67065 Strasbourg-Cedex, FRANCE. * * Tel : (33) 03 88 21 25 25 - Fax : (33) 03 88 21 25 21 * **************************************************************
Regarding my previous note...There was a typo on the phone number for Mr. Gaylan Hayes of Action Products in Virginia Beach, VA. It should be 800-865-5326 or 804-498-2623 or FAX 804-496-3894.
Ronald S. Wade, Director (I.S.P. Treasurer) Anatomical Services Division / State Anatomy Board School of Medicine, University of Maryland 655 West Baltimore St. BRB B-024 Baltimore, Maryland 21201-1559 Phone: Voice: (410) 706-3313 (410) 547-1222 (Anatomy Board) Fax: (410) 706-8107 Home: (410) 987-1869
Our annual service contract on our JEOL TEM 1210 is coming up for renewal. = I am investigating third-party service contracts and third-party demand = service. I am eager to hear other transmission microscopists=27 experience= s with service contracts, particularly third-party service.
I also invite vendors providing service in West Central Florida to contact = me via E-mail at this address (off the list).
Hello, I have a project in the plans that will require me to visualize the cuticle of a plant leaf at the TEM. I intended to cut sections via cryoultramicrotomy, and was hoping someone could suggest a reagent to stain the cuticular layer. Anyone done this? -- Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
Meeting title: "Scanning Probe Microscopy in Biomaterials Science, Dentistry and Medicine"
Aim: The aim of this meeting is to give an overview of the use of scanning probe microscopy (SPM) in biomaterials science, biotechnology, dentistry and medicine. Beginners in the field of SPM and experts will be brought together. The meeting will be rounded off with practical demonstrations.
Meeting Format: A one day meeting with invited presentations, posters and practical demonstrations.
Date 2 April 1998
Venue Redwood Lodge Hotel and Country Club, Bristol, UK.
Fees There will be a registration fee of 10 GBP per delegate to be paid on arrival at the meeting. Digital Instruments (UK) Ltd. have kindly agreed to sponsor the rest of the costs of the meeting.
Expected Audience: The meeting is aimed at a distinguished academic and industrial audience with an interest in biomaterials science, biotechnology, dentistry and medicine.
Speakers: High calibre international and national speakers. A list of speakers is available under http://www.dent.bris.ac.uk/research/materials/Speakers.html
Organisers Dr. rer. nat. Klaus D. Jandt, Mr. Richard W. Vowles (University of Bristol); Digital Instruments (UK) Ltd.
More Information/ Registration/ Programme: Can be found under http://www.dent.bris.ac.uk/spm.htm
Does anyone know of a source for light boxes comparable to the one offered by Imaging Research: Northern Light desktop illuminator? The price that they want is very high around $2000.
Peter: Just over a year ago, we switched some of our instruments to a service provider called CIC that works as an HMO. CIC has a much larger contract with the university covering all sorts of items that may need service. The motivation for the switch is that while it saves us much needed money, this arrangement is supposed to be transparent to us in the sense that we just call the usual service provider when the intrument is down, and the service provider bills CIC directly. In this way, the quality of service should not be affected. However, I already experience two shortcomings: One due to the fact that if the estimated cost of repair is above $5,000.- prior approval from CIC is required to conduct the service. If the cost is substantially higher, CIC may decide to get a "second opinion" from a repair service of their choice (imagine the delays and consequences of having many "experts" try their hands at your fine instrument!). Recently, I requested approval on a repair that was estimated at $8,600 for parts only, and got approval within a couple of days over the phone, calling CIC's toll free number. That initial repair only helped pinpoint the problem, and the instrument is still down. We are waiting for a new estimate, which in turn may lead to a request for a "second opinion" ... The other is due to the fact that instrument manufacturers will give priority to their contract customers. In our case, we put two JEOL instruments, a microprobe and a HRTEM, under CIC coverage, and continued JEOL contracts on two other JEOL instruments (wisely, our director did not put all the eggs in one basket!). Generally, we get reasonably fast service from JEOL on the last two (the instruments, not the eggs), but a little slower response on the microprobe. It was a lot harder, however, to get service for the more complex HRTEM. We called for service on the HRTEM on November 17, work started on January 8, and after some interruptions (it is still down) we are waiting for a new estimate to report to CIC. When all intruments were on JEOL contracts we were getting equally fas service.
I hope you get more information on this issue regarding other manufacturers as well as JEOL, and make an informed decision. Good luck!
Augusto
At 11:15 AM 1/26/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
} } To:histonet } From:tflore-at-lsumc.edu (Teresa Flores) } Subject:Portable/Tabletop Cryostat } } Goodmorning Histoneters. } Are any portable cryostats (tabletop) being manufactured? This portable } cryostat should be able to go from hospital to hospital. } If any have been invented, How much does it weigh? How much does it cost? } Can disposable knives be used? Any special adaptors needed? } A pathologist from Santiago, Chile has asked my boss to help him. } The Pathologist from Chile is now using a CO2 tank that attaches to a } sliding microtome which he takes from hospital to hospital. } ( I am assuming its like the one the pathologist used to frozen section a } piece of tissue, remove the section from the knife with his finger, place } the tissue on a section on a glass bowl and with a glass rod pick it up } and roll the tissue onto a paragon stain, pick tissue up and roll it onto } water, pick up the section on a slide, place a drop of glycerine, } coverslip it and view it w while my supervisor frozen sectioned another } piece of tissue (in one of the first tissue tex cryostats (in 1969)) I was } a med tech student. } Anyway, any help would be appreciate it. I have a call into Leica also. } Thanx for all you help and imput. } Teresa Flores } FAX:504-568-6037 }
My colleague is looking for software to model the crystal structure obtained after microtoming her sample. She would like to know that she has indeed found the center of a f.c.c. crystal. She wishes to identify the section relative to the entire crystal morphology.
We frequently have problems preparing materials which include soft phases such as sulphate-complexes (sulfates for the Americans). The polished sections are prone to ripping when prepared in the usual manner. We have tried various different solvents as opposed to water with varying degrees of success.
I am interested in finding out whether anybody uses special equipment or has a special technique that they use for the preparation of soft material samples.
Can anybody recommend a good image analysis course - preferably a hands on workshop type rather than purely lectures. Tropical climate and close proximity to beautiful beaches preferable.
We have recently purchased OPTIMAS and intend to utilise the system for general modal abundance phase characterisation and quality control of converter matte.
Thanks in advance
Jurgen Paetz Senior Mineralogist ARC Johannesburg sunny South Africa.
You don't mention in your posting what the .05 um powders are composed of= =2E If you have not considered microtomy as an alternative please contact us off line for a discussion of how they might be prepared for thin sectioning.
Don't be discouraged if they are hard materials, Phil Swab has published = a paper showing BN on CVD diamond on Si thin sectioned for TEM.
Steve Miller RMC Email: Steve.Miller-at-RMC-Scientific.com Website:http://www.RMC-Scientific.com/microtomes/
Hello all, Having little, (meaning 'virtually none') experience in preparing and cultivating tissue samples I was hoping someone could suggest basic reference materials/texts that might help me out here. I would be especially interested in any related work on the subject of perennial plants so that I might be of assistance to some young and aspiring botanists/microscopists.
MSA Microscopy and Microanalysis '98 Atlanta July 12 to 16
There are now only three working days to the February 1 deadline for submission of abstracts.
=85=85=85=85=85.
ICEM 14th International Congress on Electron Microscopy Cancun August 31 to September 4
Deadline for submission of abstracts is February 15 - but allow time for mailing to Mexico. Details are to be found at http://icem.inin.edu I have copies of the Second Circular and Call for Papers which I can send if you need one.
=85=85=85=85=85.
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I am looking to obtain a used TEM of late 80's - early 90's vintage. I would like EDX capabilities included, but not a necessity. If you know of such an instrument, please Email me at jfb-at-uidaho.edu.
Any smart computer folks out there who know if it is possible to run two different (ethernet and token ring) =
network cards at the same time in a Wintel platform? I'm trying to get an SEM on to two networks at the same time, or more specifically, I'm trying to get the SEM pc to act as an intermediary link=
btween an EDX (ethernet) and the lab (token ring). Thanks Bill Neill
It is possible to operate two network environments on the same machine. = What confinguration do you have, are you running, OS2, NT, 95 or 3.1?
Evex Ananlytical
----------
Any smart computer folks out there who know if it is possible to run two different (ethernet and token ring)=20 network cards at the same time in a Wintel platform? I'm trying to get an SEM on to two networks at the same time, or more specifically, I'm trying to get the SEM pc to act as an intermediary = link btween an EDX (ethernet) and the lab (token ring). Thanks Bill Neill
Could anyone share their processing cycle for human and animal nerve, and also a reliable and reproducible staining technique for thick sections? Thanks in advance for any help. Ronnie Houston Texas Scottish Rite Hospital for Children Dallas
I would think it should be possible with Windows 95 and maybe with 3.x machines. My Windows 95 network setup would allow me to allow another adapter (e.g., an IBM Token Ring card). Then the appropriate protocol would have to be loaded and assigned to that card. I could imagine Windows would get confused when it was called to do the same protocol on two different adapters. But then I know that I can have NetBeui or TCP/IP set up on both my ethernet card and my dial up adapter (i.e., modem). Normally, I would suppose you would use one protocol on your token ring network and a different one on your ethernet network.
That should allow the PC to be an intermediate stop on the way from one net to the other. You should be able to mount network drives (one from each side), and copy files back and forth while sitting at that PC. If you wanted to make the machines on one side directly visible to machines on the other, then you would need something to act as a bridge or gateway. That might require a different kind of box or more special purpose software, maybe something under Windows NT. But I will leave that to those who know more than I do.
At 12:23 PM 1/27/98 -0500, you wrote: } Any smart computer folks out there who know if it is possible to run two } different (ethernet and token ring) } network cards at the same time in a Wintel platform? } I'm trying to get an SEM on to two networks at the same time, or more } specifically, I'm trying to get the SEM pc to act as an intermediary link } btween an EDX (ethernet) and the lab (token ring). } Thanks } Bill Neill
BTW, what do you mean "Wintel" machine? Do I understand you have an ethernet link between the EDS PC and the SEM PC and that the SEM PC is also hooked up to the lab token ring? I might have hooked the EDS PC to both sides, but then our SEMs do not have full-blown PCs inside them yet anyway.
Hope this helps some. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
Dear Oliver I think the only possible way of staning the cytoskeleton of live cells is to use microinjection, but the small size of your cells makes this very difficult. I do not know much about the technique but even if you succeed in loading some cells, the injection can cause "stress" and I think small cells are more likely to react in this way. Check with experts. The alternative is labelling fixed cells. Check out "Molecular Probes" on www and search the net in general on this issue. I have just been through it myself searching for tubulin specefic dyes. I wanted to stain my cells live too, but have to live with immunolabelling of fixed cells (flagellates). You are welcome to e-mail me if you have any questions.
Best regards Jette
Note: I have no financial interest in any firms mentioned.
On Sun, 25 Jan 1998, O. Stache wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Madams and Sirs, } } I'm doing my thesis at the Aachen University of Applied Sciences. } My research involves the actin-network in human blood cells. } I've been desperately seeking for a method to stain the actin network of } lymphocytes with a fluorescence in LIVING cells, but i can't } find a method. } } Does anyone know about a method of staining the actin-network of living } cells for fluorescence mikroskopy ? } } Oliver Stache }
If you have soft samples you should be aware of this microscopy breakthrough that will give Bio-SEM labs new capabilities.
Magnetic AC Mode finally makes AFM a need tool for microscopy labs.
First Application http://green.la.asu.edu/pubs/nature/nature.html Technique http://green.la.asu.edu/pubs/APL122396/magneticprobes.html Review Article http://molec.com/educ/review1/review.html New Data acquisition http://green.la.asu.edu/pubs/MACforce/index.html http://molec.com/apps/csafm/CSAFM-app1.html
Highest resolution in situ atomic force microscopy.
Molecular imaging: - Under biological buffers - At 37C temperature control (or hot or cold) - Surface characteristics . force measurements in reaction to functionalized probes . viscoelasticity . thermal conductivity . capacitance (dielectric) . impedance . electrostatic force and surface potential . electrical conductivity . magnetic
At 03:58 PM 1/27/98 +0200, Jurgen Paetz wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Peter: } Just over a year ago, we switched some of our instruments to a service } provider called CIC that works as an HMO. CIC has a much larger contract } with the university covering all sorts of items that may need service.
*** some deleted ***
} request for a "second opinion" ... The other is due to the fact that } instrument manufacturers will give priority to their contract customers.
*** more deleted ***
This is why we have not gone with such an insurance company for our electron microscopes. The Veterinary Teaching Hospital at Colorado State University uses one for almost all their equipment, I'm told, and it works for them. We didn't want to risk it, though. When you go with an insurance company, you will not be on service contract with the manufacturer. Just my $0.02's worth.
Bonjour messieurs dames histologistes ou patologistes=0A=0AJe travaille = =E0 Lyon, France, avec mat=E9riel biologique (tissu animal) inclus en=0Ag= licol metacrilate, mais je n'ai pas microtome pour couper mes bloques de= =0Ar=E9sine. Je voudrais savoir si vous avez un microtome avec l'accessoi= re pour=0Autiliser le couteau de verre ou le couteau de diamant pour coup= es de 1-2=0Amicrons et si vous pouvez me laisser couper le mat=E9riel en = votre microtome.=0AJ'ai les couteaux. J'ai besoin de ces coupes pour ma r= echerche post-doctoral. =0AJe vous remercie.=0A=0AFrancisco Javier Hernan= dez Blazquez=0AUniversit=E9 Claude Bernard I - Lyon=0AFacult=E9 de Medici= ne=0A=0ATelefone/Fax (33) 4 78490449=0Afjhblazq-at-aol.com=0A=0A
Michael J. Lyon, Ph.D. wrote: ========================================= Does anyone know of a source for light boxes comparable to the one offered by Imaging Research: Northern Light desktop illuminator? The price that they want is very high around $2000. ========================================= Most of the suppliers of consumables and accessories for microscopy laboratories offer light boxes of varying types, sizes and costs. The light boxes offered by SPI can be found on our website.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I am sure this has come up before, but could subscribers asking for = equipment or technical assistance give a bit more information about = themselves. Someone asking for a second-hand TEM, for instance, but does not mention = where in the world they are, makes it difficult to know if some of us = out side of the USA should assist. I realise that this is the USA listserver, but there is a whole other = part of the world out side of the USA who get involved with the USA.
Thanks=20
Luc Harmsen Anaspec, South Africa Technical support for E.M. operators, world wide. anaspec-at-icon.co.za TEL: ++ 27 (0) 11 476 3455 FAX: ++ 27 (0) 11 476 7290=20
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Constance Linville expressed problems with getting thin sections to stick } together during sectioning. } } One of our applications staff suggested trying a very thin layer of a } dilute solution of rubber cement on the top and bottom faces of the block } after trimming. } } Let us all know how you do. } } (Your only problem might be getting sections apart after that!) } } Steve Miller
I am using mounting media in toluene (1:1) only in the bottom face of the block. It gives very good results!
At 07:33 PM 1/27/98 EST, Francisco Javier Hernandez Blazquez asked about microtomes for plastic (glycol methacrylate) sections.
Energy Beam Sciences manufactures the JB-4 and JB-4A microtomes for this purpose. Interested parties can find information about these instruments at our web site (http://www.ebsciences.com) or may contact me directly back-channel.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
We're preparing to do a protocol for viewing of mycorrhizal hyphae in SEM and in going over some literature we were puzzled by an instruction that called for 2 g samples of soil to be "dispersed in 200 mils of 0.1% Decon". I assume that we're not talking about rat poison here, and we've been unable to locate anything under the name Decon in the standard chemical supply catalogs.
Has anybody heard of this? Can other standard dispersal/deflocculation agents be substituted?
Thanks for any advice.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Hello All, 1. Does anyone have a solution for static electric discharge between film sheet? Using Kodak SO163, I transfer it from the film plates to a box, then off to the dark room where on occasion while separating the sheets there is a visible discharge between sheets. Sometimes quite a show but at the expense of data.
2. Not so long ago we replaced our LaB6. The image of the filament was a very nice cross with a well defined point in the center. Hi-Res work was better than ever before, in fact GREAT. The filament image held for several 2-300 hours. Then it changed overnight, blooming in the center. Ok, I thought this is awful soon & abrupt for such a failure but onward through the fog. A few days later the image became a sharp pretty well defined line. I do not believe this was an astigmatism in the optical system as I could obtain symmetrical expansion of the spot (accepting it's geometry). Last night I began to see changes to the line. I should point out that the imaging is in general still pretty good, off a bit perhaps & I have had to increase the bias voltage to get beam current up to the desired level (10uA). The instrument is a Jeol 2010 & the filament Kimball Phy. 60-06. Can anyone tell me that I have other than a filament problem? Is there a source on modes of filament failures about?
3. I thought that I understood the origin of the cross in the filament image with our Denka but the Kimball Phy. geometry is that of a sharpened cylinder. Is this due to crystal orientation rather than physical geometry?
My colleague, Xiu-lin Gao asked me to qualify the recent request made by her to the list server regarding software for modeling crystal structure as follows: I am looking for a software, which performs 3D-simulation of crystal shape and identify crystal structure of cross-section after microtoming the individual crystal from crystal surface with different cutting angles. Thanks, Lynne Garone Polaroid Corp. GaroneL-at-Polaroid.com GaoX-at-Polaroid.com
Dear Paul, There are lots of good references about preparing plant material for microscopy. Do you want to do TEM or SEM or LM? Immunolabelling? What kind of samples do you want to use? Tissue cultured cells, pieces of whole plants? If you can be a bit more specific with your request I can send you loads of references. If you're not sure right now, let me know, and I'll recommend some more general material.
Best wishes, Janine Sherrier djs56-at-mole.bio.cam.ac.uk University of Cambridge England } Hello all, } Having little, (meaning 'virtually none') experience in preparing and } cultivating tissue samples I was hoping someone could suggest basic } reference materials/texts that might help me out here. I would be } especially interested in any related work on the subject of perennial } plants so that I might be of assistance to some young and aspiring } botanists/microscopists. } } Thanks, } Paul Gerroir } }
I'm dredging up an old question, but here goes. We're looking to add a CD-Recordable unit to an existing image analysis workstation. I'd like to hear comments, good or bad, about any CD-Rs with which you've had some experience. TIA.
Regards, Bev Maleeff
SmithKline Beecham Pharmaceuticals Toxicology-US, UE 0462 709 Swedeland Road King of Prussia, PA 19406 610/270-7987 610/270-7202 fax Beverly_E_Maleeff-at-sbphrd.com
id LAA04704; Wed, 28 Jan 1998 11:44:46 -0800 Message-ID: {n1326134652.16997-at-macmail.lbl.gov}
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
All-
We have a LARGE sectioning problem. To create a display showing the internals of a TEM, we need to section the whole microscope column.
Ideally, we would like to remove a 120deg wedge, but 90deg would be acceptable (we have been told that 180deg would cause things to fall apart).
Does anyone know who might be able to carry out such a sectioning job? We would prefer someone close to Berkeley rather than have to ship the TEM too far.
Thanks for your (anticipated) help. -Mike O'Keefe (now to get back to the MSA abstract. . .).
+++++++++++++++++++++++++++ Michael A. O'Keefe, Deputy Head National Center for Electron Microscopy Lawrence Berkeley National Laboratory MS72 University of California 1 Cyclotron Road Berkeley, California 94720 tel: (510) 486-4610 fax: (510) 486-5888 email: maok-at-lbl.gov http://ncem.lbl.gov/ +++++++++++++++++++++++++++
by cosmail1.ctd.ornl.gov (PMDF V5.1-10 #23475) with ESMTP id {0ENI00M1MEXOO2-at-cosmail1.ctd.ornl.gov} for microscopy-at-Sparc5.Microscopy.com; Wed, 28 Jan 1998 14:46:38 -0500 (EST)
All-
******* LAST REMINDER !!! ********
Abstracts are due on FEBRUARY 1st (in five days!) for the Microscopy and Microanalysis '98 meeting to be held in Atlanta GA from July 12-16th, 1998. Check out the website at :
for more details on planned symposia as well as abstract submission.
Kathi
Kathleen B. Alexander Metals and Ceramics Division Oak Ridge National Laboratory P.O. Box 2008 MS-6376 Oak Ridge, TN 37831-6376 PH (423) 574-0631 FAX (423) 574-0641
} 1. Does anyone have a solution for static electric discharge between } film sheet? Using Kodak SO163, I transfer it from the film plates to a } box, then off to the dark room where on occasion while separating the } sheets there is a visible discharge between sheets. Sometimes quite a } show but at the expense of data. } Ah, yes, Kurlian microscopy. If you think it's bad using SO163, wait till you try Lo Dose! One possibility is to leave the film in the (metal?) plates and transport in a bigger box. I have often had discharge problems while putting film into the plates--it's worst in winter when the ambient air is dryest. I've found that wearing a synthetic-fiber lab coat is detrimental. The best way I've found to separate films is to hold my left thumb firmly on the stack of sheets--emulsion down, of course--and lift a corner of the top sheet with my right hand. I can then peel the sheet off the stack and release it from my thumb without causing a discharge. The secret is not to slide the top sheet over the rest of the stack. Good luck. Yours, Bill Tivol
I am seeking a student enrolled in a TEM course, interested in = collaborating on a study of spermatogenesis in a fly (Diptera) species. = I would provide the material, which is soft tissue, easily fixed, = stained, and sectioned. This would be an excellent study for someone = new to TEM, and would lead to a co-authored publication, and some very = interesting micrographs.
If you are interested, or know of someone who might be, please contact = me at:
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#c8e0d8} {DIV} {FONT color=3D#000000 size=3D2} I am seeking a student enrolled in a = TEM course,=20 interested in collaborating on a study of spermatogenesis in a fly = (Diptera)=20 species. I would provide the material, which is soft tissue, = easily fixed,=20 stained, and sectioned. This would be an excellent study for = someone new=20 to TEM, and would lead to a co-authored publication, and some very = interesting=20 micrographs. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} If you are interested, or know of = someone who=20 might be, please contact me at: {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {A=20 href=3D"mailto:srjones-at-ktc.com"} srjones-at-ktc.com {/A} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Stan Jones, = Ph.D. {/FONT} {/DIV} {/BODY} {/HTML}
Dear fellow E-Microscopists: I'm having some difficulties cutting sections with my newly achieved Nanoplast FB101 and I am wondering if there is anyone who has experience with that particular resin. My problem is, that the cured resin seems to be extremely hard and very brittle. It shatters when I attempt to cut sections of 100 nm with a diamond knife. This problem occurred after I proceeded with the embedding and hardening as follows: First I prepared MME/ catalyst mixtures using three different concentrations of the catalyst: 0.15, 0.2 and 0.25 g B52 per 10 g MME 7002. Then I pipetted my samples--unicellular algae--into the tip of a size '0' BEEM capsule, and I added just enough resin-mix to fill the cone on the tip of the capsule. Then I dehydrated the samples for two days at 40 oC and cured them for another two days at 60 oC. However, the oven went up to 80oC during the last 48hrs and maybe that caused the problem. In order to support my Nanoplast blocks, I then filled the BEEMs up with Spurr's resin that I cured overnight at 70 oC. As I already mentioned, the resin blocks turned out to be superhard and, in our experience, too brittle for cutting sections. Supposedly the resin should be 'as hard as glass', according to its developers. Is it maybe too hard by nature for cutting with a conventional diamond knive or did something go wrong? If that's the case, what went wrong? In order to find out, I'm trying to control the temperature more rigidly and to use smaller catalyst concentrations. Does anyone have different suggestions for emendations? similar experiences? ideas how to emulate the resin performance? I'd be more than happy if you could share your thoughts and ideas with me. Thanks very much, Thomas Jabusch
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Thanks to everyone who has so generously sent protocols and refrences in regaurd to my fish otolith questions. Kim
-------------------
Kim DeRuyter Histology and Electron Microscopy PO Box 755780 University of Alaska Fairbanks, Alaska 99775-5780 fnksd1-at-aurora.alaska.edu (907)-474-5452
I would like to upgrade my edx system by replacing the original Tracor Northern TN2000 processing hardware with a Pentium based system (not yet purchased). I am using Kevex model 3201-C-VS detector/2003 preamp (yes, a Kevex detector on a TN system OEM). Can someone please guide me to a reliable vendor of the best PC edx interfacing boards and software for this kind of modification? The detector will have to stay for now due to cost restrictions. This seems like the least expensive way to upgrade. What is your experience? I intend to upgrade the detector eventually.
This system is mounted on a Zeiss/LEO DSM982 Gemini FESEM.
Thank you. Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
Just had our 2 year old HP go bad. Decided to get a Dynatek, but it's not in yet. Our olympus is running strong though.
At 02:45 PM 1/28/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
} I would like to upgrade my edx system by replacing the original Tracor } Northern TN2000 processing hardware with a Pentium based system (not yet } purchased). I am using Kevex model 3201-C-VS detector/2003 preamp (yes, a } Kevex detector on a TN system OEM). Can someone please guide me to a } reliable vendor of the best PC edx interfacing boards and software for this } kind of modification?
Dear Jim and List, I too have a TN2000 with a Kevex detector, and I'd be interested in the replies. TIA. Yours, Bill Tivol
} I'm dredging up an old question, but here goes. We're looking to add a } CD-Recordable unit to an existing image analysis workstation. I'd like to } hear comments, good or bad, about any CD-Rs with which you've had some } experience. TIA.
We are happy with our 1.5 yr old Pinnacle Micro CDR burner but if I were buying another one today I would probably buy the APS brand. They make a nice compact combo unit consisting of a Jaz drive and the CD Burner for less than we paid for the Pinnacle alone.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Just a thought on soft materials and their sectioning: Several investigators have had success in the materials sciences by using cryoultramicrotomy. I think this would be worth investigating for your project as well.
Frozen ultrathin sectioning is the way to go for many samples including rubbers, plastics, polymers and other materials which may be flexible or pliable at room temperature and that do not embed well in plastic.
An additional benefit is that you can alter the sectioning properties of the material by simply changing the temperature of the cryochamber, the knife, or both. Most synthetics have fairly well-defined transition temperatures and seem to section best at or near their transition temperatures. I don't know if this would hold true for your specimens, but it is something to think about.
If you need additional information, feel free to contact me off-list.
Best regards, Bob Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Leica (Wild, Leitz, Bausch&Lomb, Cambridge, AO, Reichert-Jung) / Technical Instrument Company / American Volpi / Fostek / Stocker and Yale / AEI North America / OptiQuip / Dolan-Jenner / Osram / G.E. / Philips / Ushio / Boeckler Instruments / Heidenhain / Narishigi / Colorado Video / Visual Environments of California, Inc. / Kinetic Systems / Pacific Precision Laboratories, Inc. / Pryor Scientific / Compumotor / Sutter Instrument Co. / Advanced Database Systems / Cohu / Javeline Electronics / Optronics / Diagnostic Instruments, Inc. / Dage MTI / Hitachi / Panasonic / Polaroid / Kodak / Mitsubishi / Sony
------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. When it absolutly has to be static discharge free I'll leave the film in my dessicator overnight after removing it from the camera. It seems to me that the film experiences this problem most often right out of the microscope where it has been exposed to the most dry, dessicated conditions that film will ever see. Overnight in the film dessicator (rotary pumped chamber) and the film is less apt to have static discharges. Hope this helps. Greg Strout
} } Dear Bruce, } } } 1. Does anyone have a solution for static electric discharge between } } film sheet? Using Kodak SO163, I transfer it from the film plates to a } } box, then off to the dark room where on occasion while separating the } } sheets there is a visible discharge between sheets. Sometimes quite a } } show but at the expense of data. } } } -- ======================================================== Greg Strout Electron Microscopist, University of Oklahoma e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not neccessarily those of the University of Oklahoma ========================================================
} Hello All, } 1. Does anyone have a solution for static electric discharge between } film sheet? Using Kodak SO163, I transfer it from the film plates to a } box, then off to the dark room where on occasion while separating the } sheets there is a visible discharge between sheets. Sometimes quite a } show but at the expense of data.
This generally seems to happen more low-humidity environments. You might try adjusting the humidity of your work space, if possible. You might also try working with your films on a grounded anti-static mat, like those made for working with sensitive electronic components and computer parts. Don't know what they cost, but they seem to help. Also, m-o-v-e s-l-o-w when working with films. Shuffling them around fast is guaranteed to generate lightning storms. } } 2. Not so long ago we replaced our LaB6. The image of the filament was a } very nice cross with a well defined point in the center. Hi-Res work } was better than ever before, in fact GREAT. The filament image held for } several 2-300 hours. Then it changed overnight, blooming in the } center. Ok, I thought this is awful soon & abrupt for such a failure } but onward through the fog. A few days later the image became a sharp } pretty well defined line. I do not believe this was an astigmatism in } the optical system as I could obtain symmetrical expansion of the spot } (accepting it's geometry). Last night I began to see changes to the } line. I should point out that the imaging is in general still pretty } good, off a bit perhaps & I have had to increase the bias voltage to get } beam current up to the desired level (10uA). The instrument is a Jeol } 2010 & the filament Kimball Phy. 60-06. } Can anyone tell me that I have other than a filament problem? Is } there a source on modes of filament failures about? }
Just curious---is your filament voltage stable? (I don't mean accelerating voltage here.) As I recall, LaB6 filaments come with a maximum voltage written on the box, determined by the manufacturer. On some Hitachi scopes, voltage can be set on the configuration menu and has a big effect on how saturation and bias affect the filament image. You can adjust this if, for example, you're getting a shadow at what should be full saturation. Take great care not to set it higher than the manufacturer's recommendation, however, or you can damage the crystal. } } Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
1) A new season of MSA Bulletins is about to begin. As you have all been going through your negatives looking for the perfect images to include in your abstracts - see 3 - this is a good opportunity to remind you that the Bulletin is always looking for fun, interesting, humerous, witty, etc. images to liven up the content. If you have any such material in almost any form, they can be sent to me at the address at the end of this message.
2) Those of you who are MSA members should have recieved a Project Micro special issue with your recent copy oc Microscopy and Analysis.
The Minnesota Microscopy Society web page has recently been updated for those of you interested in showing kids (of any age) the sort of thing that microscopes can do for you can check out our mi=onthly exploration at:
3) Obligatory reminder (as program vice-chair elect):
__________________________________________________________________________ Abstracts are due on FEBRUARY 1st (this weekend!) for the Microscopy and Microanalysis '98 meeting to be held in Atlanta GA from July 12-16th, 1998. Check out the website at :
for more details on planned symposia as well as abstract submission.
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
} } 1. Does anyone have a solution for static electric discharge between } } film sheet? Using Kodak SO163, I transfer it from the film plates to a } } box, then off to the dark room where on occasion while separating the } } sheets there is a visible discharge between sheets. Sometimes quite a } } show but at the expense of data. } } } Ah, yes, Kurlian microscopy. If you think it's bad using SO163, } wait till you try Lo Dose! One possibility is to leave the film in the } (metal?) plates and transport in a bigger box. } I have often had discharge problems while putting film into the } plates--it's worst in winter when the ambient air is dryest. I've found } that wearing a synthetic-fiber lab coat is detrimental. The best way I've } found to separate films is to hold my left thumb firmly on the stack of } sheets--emulsion down, of course--and lift a corner of the top sheet with } my right hand. I can then peel the sheet off the stack and release it } from my thumb without causing a discharge. The secret is not to slide } the top sheet over the rest of the stack. Good luck.
A shocking state of affairs. I got a charge out of this message.
I hearily second W. Tivol's recommendations - especially about NOT sliding one sheet of film over another. In addition, we do the following:
1. Work on top of a grounded surface. We use carbon-impregnated 3 x 3 ft mats attached to a grounding strip. Computer people use them in order to work on electronic components that are sensitive to static. It is also a good idea (in high static situations) to wear a grounding strap on your wrist -- again just like the computer types. All negs are transported to and from the strip in metal containers so that when one places the container down on the strip, the static is discharged from the exterior of the container onto the mat.
2. The area around the mat is wet wiped with a lintless or cotton towel in order to generate a humid environment nearby. This really helps. You should take care, however, not to place any of your films or containers onto the moist zones. Work on the mat. If you suspect static buildup, you can dissipate the charge by breathing onto the film prior to handling - -} like you were trying to fog your glasses in order to clean them.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Over the years, and today too, I have learned that when taking polarized light microscopy micrographs on Ektachrome 64T there is a color shift depending on how much the polarizer and analyzer are crossed; i.e. if the polarizer and analyzer are parallel to each other, then there is little color shift in the color slide. However, as the polarizer and analyzer are brought to the crossd (perpendicular) position, the background takes on a very definite green hue, almost as if the filters are selectively removing the blue and red portions of the spectrum.
Can anyone explain to me why these two supposedly neutral filters when brought nearly to the crossed position, say within 6-12 degrees of being crossed, cause the micrograph to take on a green cast that requires a 15M to make the background a more neutral gray?
Thanks in advanace for explaining either a very complex or very simple question. {As long as the answer is simple, I'll be OK :-)}
Damian Neuberger neuberd-at-baxter.com or dneuberger-at-mindspring.com
At 10:33 PM 1/28/98 -0600, you wrote: } Randy, } The only feed back I would have on the filament heater current would be in the } beam current readout. Beam current is very steady. Thanks for the suggestion, } I'll prod the field engineer when he arrives. With respect to saturation, the } image is reasonable, just constantly evolving geometry. } } Peter,
I'm not talking about the filament heating current, but rather the voltage applied to the filament. I'm not an electronics guru, nor am I familiar with JEOL scopes, but with regard to the filament, you have filament current, filament voltage, gun bias, and accelerating voltage, and they are all different. I don't know much more than that, unfortunately. The only LaB6 I've worked with was on a Hitachi H7100FE, and as long as we didn't exceed the recommended voltage supplied by Hitachi, we could often get rid of unwanted filament shadows and weird shapes seen at cross-over. These things definitely changed as the filament aged. Wish I was more informed. Please let me know what you find out, because I'm curious, too.
One final thought. When we have seen really strange sights at crossover, like straight lines, etc., very often the condenser stigmators were way off. You might check this. If you can't get decent stigmation, look into some kind of contamination in the gun area or on one of the column apertures. A chunk of something nasty in an aperture can really El Nino your day, if you catch my drift.
Best of luck and let me know what happens.
Randy
Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
I'm sorry I don't recall the name of the gentleman responsible, but at least a few EMs were sectioned here in the midwest a couple of decades ago. I have seen the results both at the Museum of Science and Industry in Chicago and at the former Center for Electron Microscopy at the University of Illinois. I have no contacts for the Museum, but either Dr. Richard Crang or Dr. Birute Jakstys, who are both I believe still with the University of Illinois, could possibly give further information.
As I recall, the first step was to stabilize the internal electron optic column parts by vacuum impregnation of a plastic or epoxy. After all of the internal spaces were filled, a simple series of longitudinal saw cuts were made to section the instrument followed by a polishing of the exposed surfaces. With attention to detail, you could probably manage to have all of the work done locally. It may be easier to do the impregnation in-house and use a good local contract machine shop for the actual sectioning and surface finishing. With the impregnation, a 180 degree cut should not be a problem, and the complementary section could perhaps be donated to a local museum.
If you have any problems, or find the project too daunting, let me know. I could certainly complete the project here, at a price ;)
} All- } } We have a LARGE sectioning problem. To create a } display showing the internals of a TEM, we need } to section the whole microscope column. } } Ideally, we would like to remove a 120deg wedge, } but 90deg would be acceptable (we have been told } that 180deg would cause things to fall apart). } } Does anyone know who might be able to carry out } such a sectioning job? We would prefer someone } close to Berkeley rather than have to ship the } TEM too far. } } Thanks for your (anticipated) help. } -Mike O'Keefe } (now to get back to the MSA abstract. . .). } } +++++++++++++++++++++++++++ } Michael A. O'Keefe, Deputy Head } National Center for Electron Microscopy } Lawrence Berkeley National Laboratory MS72 } University of California } 1 Cyclotron Road } Berkeley, California 94720 } tel: (510) 486-4610 } fax: (510) 486-5888 } email: maok-at-lbl.gov } http://ncem.lbl.gov/ } +++++++++++++++++++++++++++ } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
} Date: Thu, 29 Jan 1998 19:33:17 +1000 } To: Beverly_E_Maleeff-at-sbphrd.com } From: Wolf Schweitzer {wschweitzer-at-access.ch} } Subject: Re: Looking for opinions on CD-R } Cc: } Bcc: } X-Attachments: } } The Philips CDD 2660 seems to work alright as a writer/reader device. I } use Apple Macintosh System 7.6.1 and Astarte Toast Software which came } with the lot, which seems to be sold by Adaptec www.adaptec.com, nowadays. } } However, the Philips CD-recordables I purchased at a regular price (not a } cheap offer), and used them for Data Backup, have now 6-9 months after } purchase/burning, unfortunately developed several (about 2 to 25 per } CD-recordable) small, unrecoverable (hardware)-holes (degradation ? } contamination ?) of up to 0,5-1,0 mm diameter in the GREEN SURFACE which } as of my knowledge, holds the data. NOT affected under completely (sic!) } SIMILAR storage/use conditions were CD-recordables manufactured by } Traxdata, Kodak, Ritek and 3M. } } All of my PHILIPS-CD-recordables have been affected, in different degree } according to their age. They also have stopped working and lost data in } different amounts, which gave me the clue that the green surface holes } might have something to do with it. There existed a correlation between } the degree of dysfunction and the number of hole present. } } A request to the manufacturer, Philips, by E-Mail, has been unanswered up } to now, so I assume they consider themselves, or me, dead in this sector } anyway. Perhaps it is their attitude, which prevents them of at least } sending an excuse. } } It has cost me considerable time to recover most/some of the data off } these Philips CD-recordables, where the Philips CD-writer/reader CDD 2660 } proved to be more error tolerant than the internal Power Mac 8600 CD } reader. So I was quite happy to have this company's hardware, though. } Unfortunately, the Macintosh was crashed so many times while trying to } recover the data, it was f**** up totally, I had to completely reinstall } the complete system /applications / data, which were fortunately on Kodak } CD-recordable by then. } } It appears to be necessary to regularly (2-3 months) check the } CD-recordables, and make double/triple burns on CD-rec'ables of different } manufacturers in order to avoid unpleasant surprises. Also, I will be } going for a TAPE backup as soon as I get the money to get a good one, as } DAT tapes still seem to be more reliable and cheaper on the long run. } } Perhaps you search for CD FAQ on Internet, using some sophisticated Web } Crawler. There are some pretty desillusionizing ( {---?grammar) views out } there. } } So long, } Wolf } } }
------------------------------------------------------------------------ Wolf Schweitzer, MD, Swiss Research Fellow, mailto:wschweitzer-at-access.ch Victorian Institute of Forensic Medicine, Southbank/Melbourne, Australia
A new creature on the market, these organizations seek to justify themselves by providing a reduction in service costs by reducing the 'insurance' inherent in a service contract and by trying to artificially increase competition. They basically work by providing service on a billable, rather than contract, basis. They will use whatever service provider they deem reasonable and cheap. When you sign up with them, you are turning over to them any and all decisions over who will actually provide service on your instrument and what time frame the service will be performed in.
While this approach might be acceptable for health related expenses (certainly open to conjecture) - for a field as esoteric as analytical instrumentation, it really doesn't hold up. There simply aren't that many service providers available to foster the competative pricing that this approach requires. In the health field, there is sufficient quantity and profit margin to warrant third party competition in areas such as MRI, CAT and PET service. That is not true in analytical instrumentation.
The analytical service organizations that you will find are generally small businesses that are in business because they enjoy the instruments and are disenfranchised by the manufacturers. While there is considerable headroom in the profit margins of analytical instrument manufacturers, there is a lack of third party service providers. That being said, you should still be able in most cases to reduce your maintenance costs by 20% by considering third party sources.
The 'HMOs' will, however, protect themselves against any potential large scale problems, more so than the small providers. I am, of course, biased by the nature of my business, but I feel that if you aggressively seek out a third party source of service, and intelligently decide for yourself who can provide the best service at the most reasonable price, you will be able to reduce your costs considerably while ensuring a reasonable level of service.
In support of this, be aware that most small service providers are much more flexible in their contract services, and can usually provide services tailored to your maintenance needs - i.e. can adjust their contracts to your requirements for preventative maintenance, emergency service and replacement parts needs. Like most service providers, I have standard service packages. However, like most independent service providers, I am always willing to provide customized contracts designed to reduce your maintenance costs.
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } -------------------------------------------------------------------- } ---. } } Peter: } Just over a year ago, we switched some of our instruments to a } service provider called CIC that works as an HMO. CIC has a much } larger contract with the university covering all sorts of items that } may need service. The motivation for the switch is that while it } saves us much needed money, this arrangement is supposed to be } transparent to us in the sense that we just call the usual service } provider when the intrument is down, and the service provider bills } CIC directly. In this way, the quality of service should not be } affected. However, I already experience two shortcomings: One due } to the fact that if the estimated cost of repair is above $5,000.- } prior approval from CIC is required to conduct the service. If the } cost is substantially higher, CIC may decide to get a "second } opinion" from a repair service of their choice (imagine the delays } and consequences of having many "experts" try their hands at your } fine instrument!). Recently, I requested approval on a repair that } was estimated at $8,600 for parts only, and got approval within a } couple of days over the phone, calling CIC's toll free number. That } initial repair only helped pinpoint the problem, and the instrument } is still down. We are waiting for a new estimate, which in turn may } lead to a request for a "second opinion" ... The other is due to } the fact that instrument manufacturers will give priority to their } contract customers. In our case, we put two JEOL instruments, a } microprobe and a HRTEM, under CIC coverage, and continued JEOL } contracts on two other JEOL instruments (wisely, our director did } not put all the eggs in one basket!). Generally, we get reasonably } fast service from JEOL on the last two (the instruments, not the } eggs), but a little slower response on the microprobe. It was a lot } harder, however, to get service for the more complex HRTEM. We } called for service on the HRTEM on November 17, work started on } January 8, and after some interruptions (it is still down) we are } waiting for a new estimate to report to CIC. When all intruments } were on JEOL contracts we were getting equally fas service. } } I hope you get more information on this issue regarding other } manufacturers as well as JEOL, and make an informed decision. Good } luck! } } Augusto } } } At 11:15 AM 1/26/98 -0500, you wrote: } } ------------------------------------------------------------------- } } ----- The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } ------------------------------------------------------------------- } } ----. } } } } Our annual service contract on our JEOL TEM 1210 is coming up for } } renewal. } I am investigating third-party service contracts and third-party } demand service. I am eager to hear other transmission } microscopists' experiences with service contracts, particularly } third-party service. } } I also invite vendors providing service in } West Central Florida to contact me via E-mail at this address (off } the list). } } Thanks, } } Peter O. Steele, Ph.D., PMIAC } } E-mail: } steelep-at-allkids.org } } } ____________________________________________________________________ } _____ Augusto A. Morrone 107D-MEL, P.O. Box 116400 } MAIC Materials Science and Engineering } University of Florida } Gainesville, FL 32611 } (352) 392-1497 or 6985 } Fax: (352) 392-0390 } amorr-at-mse.ufl.edu } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Decon is the registered trade mark of Decon Laboratories Ltd and in this case probably refers to Decon liquid which is a general purpose surfactant used for cleaning glassware in biological and radioactive work. It can be ordered through Fisher Scientific. Hope this helps.
Dan Hill Department of Biochemistry Cambridge University UK
On Wed, 28 Jan 1998, Randy Tindall wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings, } } We're preparing to do a protocol for viewing of mycorrhizal hyphae in SEM } and in going over some literature we were puzzled by an instruction that } called for 2 g samples of soil to be "dispersed in 200 mils of 0.1% Decon". } I assume that we're not talking about rat poison here, and we've been } unable to locate anything under the name Decon in the standard chemical } supply catalogs. } } Has anybody heard of this? Can other standard dispersal/deflocculation } agents be substituted? } } Thanks for any advice. } } Randy } } } Randy Tindall } Electron Microscope Laboratory } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } } rtindell-at-nmsu (work) } nrtindall-at-zianet.com (home) }
On Wed, 28 Jan 1998 Beverly_E_Maleeff-at-sbphrd.com wrote: } } I'm dredging up an old question, but here goes. We're looking to add a } CD-Recordable unit to an existing image analysis workstation. I'd like to } hear comments, good or bad, about any CD-Rs with which you've had some } experience. TIA. } } } Regards, } Bev Maleeff
We are using a sony 2X with the Easy CD-Pro 1.5 software. It is OK.
We too have purchased the Philips CDD 2600. With alittle more than a = years worth of operations it continue to function as expected. But on = the other hand, we have upgraded to the Adaptec Easy Creator version = 3.0. We find this software to be easyier to use and compatible with = Windows NT 4.0.
Peter Tarquinio Evex Ananlytical www.evex.com
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} Date: Thu, 29 Jan 1998 19:33:17 +1000 } To: Beverly_E_Maleeff-at-sbphrd.com } From: Wolf Schweitzer {wschweitzer-at-access.ch} } Subject: Re: Looking for opinions on CD-R } Cc: } Bcc: } X-Attachments: } } The Philips CDD 2660 seems to work alright as a writer/reader device. I } use Apple Macintosh System 7.6.1 and Astarte Toast Software which came } with the lot, which seems to be sold by Adaptec www.adaptec.com, = nowadays. } } However, the Philips CD-recordables I purchased at a regular price (not = a } cheap offer), and used them for Data Backup, have now 6-9 months after } purchase/burning, unfortunately developed several (about 2 to 25 per } CD-recordable) small, unrecoverable (hardware)-holes (degradation ? } contamination ?) of up to 0,5-1,0 mm diameter in the GREEN SURFACE = which } as of my knowledge, holds the data. NOT affected under completely = (sic!) } SIMILAR storage/use conditions were CD-recordables manufactured by } Traxdata, Kodak, Ritek and 3M. } } All of my PHILIPS-CD-recordables have been affected, in different = degree } according to their age. They also have stopped working and lost data in } different amounts, which gave me the clue that the green surface holes } might have something to do with it. There existed a correlation between } the degree of dysfunction and the number of hole present. } } A request to the manufacturer, Philips, by E-Mail, has been unanswered = up } to now, so I assume they consider themselves, or me, dead in this = sector } anyway. Perhaps it is their attitude, which prevents them of at least } sending an excuse. } } It has cost me considerable time to recover most/some of the data off } these Philips CD-recordables, where the Philips CD-writer/reader CDD = 2660 } proved to be more error tolerant than the internal Power Mac 8600 CD } reader. So I was quite happy to have this company's hardware, though. } Unfortunately, the Macintosh was crashed so many times while trying to } recover the data, it was f**** up totally, I had to completely = reinstall } the complete system /applications / data, which were fortunately on = Kodak } CD-recordable by then. } } It appears to be necessary to regularly (2-3 months) check the } CD-recordables, and make double/triple burns on CD-rec'ables of = different } manufacturers in order to avoid unpleasant surprises. Also, I will be } going for a TAPE backup as soon as I get the money to get a good one, = as } DAT tapes still seem to be more reliable and cheaper on the long run. } } Perhaps you search for CD FAQ on Internet, using some sophisticated Web } Crawler. There are some pretty desillusionizing ( {---?grammar) views = out } there. } } So long, } Wolf } } }
------------------------------------------------------------------------ Wolf Schweitzer, MD, Swiss Research Fellow, mailto:wschweitzer-at-access.ch Victorian Institute of Forensic Medicine, Southbank/Melbourne, Australia
Assign TCP/IP address to the client machines, assign TCP/IP address the = primary server machine that will coordinate the client machines. =20 Install a the token ring card in the server machine, and assign it the = TCP/IP address for the token ring network. This is most efficiently = accomplished with Windows NT.
Peter Tarquinio Evex Analytical www.evex.com
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Any smart computer folks out there who know if it is possible to run two different (ethernet and token ring)=20 network cards at the same time in a Wintel platform? I'm trying to get an SEM on to two networks at the same time, or more specifically, I'm trying to get the SEM pc to act as an intermediary = link btween an EDX (ethernet) and the lab (token ring). Thanks Bill Neill
FWIW, My HP has been working well. Thought about purchasing a Phillips, since it offered additional record mode, but price won. A couple of notes:
Compare software bundles and the various write modes allowed by the hard/software combination.
Beware of system performance requirements. For example, to copy another CD (without first moving contents to a HDD) a fast SCSI CD rom is usually required.
As the CD-R write speed increases, the demand on the source HDD can be significant. Slow IDE systems may cause buffer underuns (in such a case, you just made a frisbe). Safer ( and costly) is a fast SCSI HDD. Along the same line, the more buffer in the CD-R, the better. Most have 1 MB, some more.
Woody White, Electron Microscopist
McDermott Technology, Inc. http://www.mtiresearch.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To: microscopy-at-msa.microscopy.com-at-inet cc:
Colleagues:
I'm dredging up an old question, but here goes. We're looking to add a CD-Recordable unit to an existing image analysis workstation. I'd like to hear comments, good or bad, about any CD-Rs with which you've had some experience. TIA.
Regards, Bev Maleeff
SmithKline Beecham Pharmaceuticals Toxicology-US, UE 0462 709 Swedeland Road King of Prussia, PA 19406 610/270-7987 610/270-7202 fax Beverly_E_Maleeff-at-sbphrd.com
by smtp.uky.edu (8.8.8/8.8.5) with ESMTP id IAA01805; Thu, 29 Jan 1998 08:25:16 -0500 (EST) Received: from naresh (naresh.cffls.uky.edu [128.163.22.30]) by pop.uky.edu (8.8.4/(UKY.POP.1.4)) with SMTP id IAA04847; Thu, 29 Jan 1998 08:20:07 -0500 (EST) Received: by localhost with Microsoft MAPI; Thu, 29 Jan 1998 08:18:51 -0500 Message-ID: {01BD2C8E.87C27C10.naresh-at-pop.uky.edu} {microscopy-at-Sparc5.Microscopy.Com}
Just yesterday, I noticed an ad in Microscopy Today (issue #98-1) by Noran (formerly Tracor Northern) announcing digital microanalysis system for windows NT. May be they can give you a reasonable trade-in value for your TN2000.
-----Original Message-----
I would like to upgrade my edx system by replacing the original Tracor Northern TN2000 processing hardware with a Pentium based system (not yet purchased). I am using Kevex model 3201-C-VS detector/2003 preamp (yes, a Kevex detector on a TN system OEM). Can someone please guide me to a reliable vendor of the best PC edx interfacing boards and software for this kind of modification? The detector will have to stay for now due to cost restrictions. This seems like the least expensive way to upgrade. What is your experience? I intend to upgrade the detector eventually.
This system is mounted on a Zeiss/LEO DSM982 Gemini FESEM.
Thank you. Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
The color filtering with nearly crossed "plastic" polarizers (where the birefringent material that is separating the light into two polarized rays and losing one) will depend on the type and quality of the material and its history. Edmund Scientific offers both brown and gray polarizers. I don't know if crossed Nicols (calcite) give colors.
Leonard Corwin Fort Dodge Animal Health, Animal Health Research Analytical Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
There are three RMC TEMs cross sectioned now: - AFIP Medical Museum, Wash. D.C. - Museum of Science and Industry, Chicago - University of Illinois, Urbana, Center for Electron Microscopy (at leas= t it was a year ago)
These were all done Raymond J. Miller (my father), I helped only occasionally so I don't know all the details. Here are some.
It took up to FIVE columns to get a single good one. The lenses were drilled and tapped, top and bottom, high pressure fittings were threaded in. A combination of pressure and vacum was used to infiltrate the thousands of turns of very fine wire. =
Ray. ended up going to the president of DoAll saw company to get help wit= h the cross sectioning. The problem is that the very soft copper wire pulls=
out easily and leaves voids. It is my recollection that Ray ended up usin= g extremely slow speeds with the finest tooth blades available. =
If you wish more information please contact me off line, Steve Miller, =
There is a discussion of this phenomenon, called "Green Disease" on p. 85-87 in "Polarized Light Microscopy", by W. C. McCrone, L. B. McCrone, and J. G. Delly, Ann Arbor Science Michigan, 1978. The problem is attributed to a reciprocity failure of the film even in the range of normal exposure times. This in turn, is attributed to cases where the light intensity ratio of the brightest to darkest objects exceeds 3:1. This means that the film may work well for brightfield situations, but will work poorly in crossed polarized light situations. I am no expert on this, so I refer you to the reference. Several brands of film are discussed. Hope this helps.
} To All, } } Over the years, and today too, I have learned that when taking polarized } light microscopy micrographs on Ektachrome 64T there is a color shift } depending on how much the polarizer and analyzer are crossed; i.e. if the } polarizer and analyzer are parallel to each other, then there is little } color shift in the color slide. However, as the polarizer and analyzer are } brought to the crossd (perpendicular) position, the background takes on a } very definite green hue, almost as if the filters are selectively removing } the blue and red portions of the spectrum. } } Can anyone explain to me why these two supposedly neutral filters when } brought nearly to the crossed position, say within 6-12 degrees of being } crossed, cause the micrograph to take on a green cast that requires a 15M } to make the background a more neutral gray? } } Thanks in advanace for explaining either a very complex or very simple } question. {As long as the answer is simple, I'll be OK :-)} } } Damian Neuberger } neuberd-at-baxter.com } or } dneuberger-at-mindspring.com
Cynthia J. Zeissler Physical Scientist National Institute of Standards and Technology cynthia.zeissler-at-nist.gov 301-975-3910
I used to have similar problem with Eastman positive release film 5302 (35 mm) when the bulk film was stored in a desiccator. The problem was due to dryness. We just leave the bulk film in atmospheric condition these days.
We also use Kodak SO163 but never experience this problem. Perhaps you can leave the films in the dark room for a while so that they can pick up some moisture before doing processing.
Hope this helps
Ann Fook ========================================== } } 1. Does anyone have a solution for static electric discharge between } } film sheet? Using Kodak SO163, I transfer it from the film plates to a } } box, then off to the dark room where on occasion while separating the } } sheets there is a visible discharge between sheets. Sometimes quite a } } show but at the expense of data. } } } -- ========================================================
} } } } The Philips CDD 2660 seems to work alright as a writer/reader device. I } } use Apple Macintosh System 7.6.1 and Astarte Toast Software which came } } with the lot, which seems to be sold by Adaptec www.adaptec.com, nowadays. } } } } However, the Philips CD-recordables I purchased at a regular price (not a } } cheap offer), and used them for Data Backup, have now 6-9 months after } } purchase/burning, unfortunately developed several (about 2 to 25 per } } CD-recordable) small, unrecoverable (hardware)-holes (degradation ? } } contamination ?) of up to 0,5-1,0 mm diameter in the GREEN SURFACE which } } as of my knowledge, holds the data. NOT affected under completely (sic!) } } SIMILAR storage/use conditions were CD-recordables manufactured by } } Traxdata, Kodak, Ritek and 3M. } }
} } manufacturers in order to avoid unpleasant surprises. Also, I will be } } going for a TAPE backup as soon as I get the money to get a good one, as } } DAT tapes still seem to be more reliable and cheaper on the long run.
I am using the same CD-Writer with my Power Macintosh 7500 and System 8.1 at home for burning CR-recordables using Astarte Toast. Now after about half a year after purchase, the only problems I had from time to time were errors reported by the drive and subsequent loss of that CD's which were due to the brand of CD used. Thats my explanation for the moment, because after I switched from Imation CD's (cheap, green) to Kodak (more expensive, golden) and also altered my SCSI chain, I had no more problems.
You say you want to go for tape (DAT I assume): That's were I came from and it was hopeless, with a drive at home connected to the macintosh and also with a drive of a SGI workstation in the institute !
Christian
Dipl Biol Christian Zuppinger ETH Hoenggerberg Institut fuer Zellbiologie, HPM F27 CH-8093 Zurich Switzerland
} Can anyone explain to me why these two supposedly neutral filters when } brought nearly to the crossed position, say within 6-12 degrees of being } crossed, cause the micrograph to take on a green cast that requires a 15M } to make the background a more neutral gray?
Yep. John Delly can. Although he wrote the following paragraphs some years ago, "the more things change, the more they stay the same." Here's what he had
to say about reciprocity failure and "green disease" in the Particle Atlas:
"Reciprocity failure
"The reciprocity law states that the exposure density is proportional to the intensity times the duration. Therefore, equivalent exposure densities should be obtained by proportionately increasing the exposure time with a lower light intensity. Reciprocity failure means that equivalent exposure densities are not
obtained for a given intensity and exposure time. This failure generally occurs with exposures less than one one-thousandth of a second or greater than one second. Such short exposures are not usually encountered in photomicrography, but long exposures are quite common. These require not only
greater than calculated exposure times but also additional color-correcting filters. Most films require color-correction filters with exposure times greater than one second. When anticipated exposure times exceed 10 seconds, it
is best to choose a more intense light source to avoid sample vibration and the
lengthy procedure of selecting the proper color-correcting filters.
" 'Green disease'
"Photomicrographers face an uncommon problem when using slightly uncrossed polars. They get a bluish-green to green background on the transparency, instead of the gray observed in the microscope field. The problem is related to reciprocity failure even though it occurs with normal exposures of 1/10 to 1/124 sec.
"Most color films cannot accurately reproduce all colors if the light intensity
ratio of the brightest to the darkest objects exceeds 3:1. If this ratio is exceeded, exposures correct for the bright objects will result in underexposed,
"off-color" dark objects. With slightly uncrossed polars, the intensity ratio of interference colors to background is often as great as 15:1. Since our exposures are chosen to be correct for the interference colors of anisotropic particles, the background is underexposed and off-color green instead of gray. Transparencies of colorless isotropic particles that appear green cannot be used as reference standards or as records of visual microscopy. The appearance
of colored isotropic particles is also masked or altered. Anisotropic particles, however, show correct colors within the limits of film fidelity. These green backgrounds have been called the 'green disease.' "
Thanks John. And now you know, Damian.
(For more information about the Particle Atlas Electeonic Edition, email me at sshaffer-at-microdataware.com .)
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
Can anybody tell us, what is the best way to fix and embedd tabacco seeds for transmission electron microscopy? We tried 2-times without success. One of these was by using seeds which were pierced with a needle to get better penetration. We used relative long times for dehydration/embedding. Probably seeds were not fixed and/or lipid not penetrated by the embedding medium. The lipid came out during sectioning. Do we have to cut them completely to get penetration? If yes, how to avoid the swelling and thus disturbing of parts of the seeds? Special procedures? Is Epon better because of the lipids? I have no experience with seed embeddings - unfortunately.
Arthur Dr. Arthur Schuessler University of Heidelberg Zellenlehre Im Neuenheimer Feld 230 D-69120 Heidelberg Germany
Stuart - Wow! I've just looked at the "Gallery" that you described on the listserver earlier today. I guess you Minnesotans are so quiet because you're busy DOING things. It is by far the best outreach image collection I've seen on the web, and I've looked at a lot of them. So many are just "gee whiz" collections of images, without a progression of magnifications and clear explanation. I hope that you'll consider publishing the whole collection as a MSA Bulletin issue; it would be great for use in microscopy outreach. We aren't yet in a world with a computer in every classroom, or a color printer down the hall...
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Ah, yes, the TN2000 with a Kevex detector! We used to have one of those on a JEOL JSM-U3. That was before Tracor did their own detectors.
I can't say for sure, but I would suppose that the system is a good candidate for an upgrade. Ours used a Kevex pulse processor in the Tracor chassis.
We recently upgraded a Kevex Delta to an IXRF Systems unit. We kept the detector, pre-amp, and pulse processor from the old system. The box from IXRF included the MCA electronics and the necessary interface to the PC. The price for our fairly full-blown system upgrade was under $40,000 including the cost of the Pentium PC.
Disclaimer: In addition to IXRF, there are other good systems that retain your old detector and electronics. I happen to be pleasantly biased since we went with IXRF.
We bought At 03:53 PM 1/28/98 -0400, you wrote: } } I would like to upgrade my edx system by replacing the original Tracor } Northern TN2000 processing hardware with a Pentium based system (not yet } purchased). I am using Kevex model 3201-C-VS detector/2003 preamp (yes, a } Kevex detector on a TN system OEM). Can someone please guide me to a } reliable vendor of the best PC edx interfacing boards and software for this } kind of modification? The detector will have to stay for now due to cost } restrictions. This seems like the least expensive way to upgrade. What is } your experience? I intend to upgrade the detector eventually. } } This system is mounted on a Zeiss/LEO DSM982 Gemini FESEM. } } Thank you. } Jim } } James S. Romanow ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
ANS offers a complete line of upgrade solutions for all manufacturers systems. For those interested please contact sales-at-ansxray.com or visit our web site at www.ansxray.com. ************************************************* * Bill Hardy, President * * American Nuclear Systems, Inc. * * 1010 Commerce Park Dr., Suite G * * Oak Ridge, TN 37830-8026 * * (800) 980-9284 FAX: (423) 482-6253 * * www.qtmsys.com Email: bhardy-at-qtmsys.com * *************************************************
} ... Now after about } half a year after purchase, the only problems I had from time to time } were errors reported by the drive and subsequent loss of that CD's which } were due to the brand of CD used. Thats my explanation for the moment, } because after I switched from Imation CD's (cheap, green) to Kodak (more } expensive, golden) and also altered my SCSI chain, I had no more problems. } ...
Your point about altering your SCSI chain reminds me of a period of trouble-shooting various problems on several computers ... In the end I realized high quality cables should be considered as part of any initial SCSI investment ... you won't be sorry. cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility at the University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
} } Date: Thu, 29 Jan 1998 19:33:17 +1000 } } To: Beverly_E_Maleeff-at-sbphrd.com } } From: Wolf Schweitzer {wschweitzer-at-access.ch} } } Subject: Re: Looking for opinions on CD-R } } Cc: } } Bcc: } } X-Attachments: } } } } The Philips CDD 2660 seems to work alright as a writer/reader device. I } } use Apple Macintosh System 7.6.1 and Astarte Toast Software which came } } with the lot, which seems to be sold by Adaptec www.adaptec.com, nowadays. } } } } However, the Philips CD-recordables I purchased at a regular price (not a } } cheap offer), and used them for Data Backup, have now 6-9 months after } } purchase/burning, unfortunately developed several (about 2 to 25 per } } CD-recordable) small, unrecoverable (hardware)-holes (degradation ? } } contamination ?) of up to 0,5-1,0 mm diameter in the GREEN SURFACE which } } as of my knowledge, holds the data. NOT affected under completely (sic!) } } SIMILAR storage/use conditions were CD-recordables manufactured by } } Traxdata, Kodak, Ritek and 3M. } }
To all CD-R users
Please read a series of articles on CD-R by Stephen St. Croix in Mix magazine (a pro audio publication) October, February, March and July 1994 issues (July is the most informative). He has researched the different reactive-dyes used in making CD-Rs. The green cyanine can be very unreliable (1 year shelf life, 5 year data storage life). Do not leave them in the sunlight ever. Use the yellow-gold phthalocyanine disks instead (100 year claimed shelf and data life).
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
The newer 4pi SE-II boards, although developed for the Mac, are PCI bus boards.
I've only used the Mac versions, but their FAQ page at {http://www.4pi.com/information/faqs/generalfaq.shtml#eds} says:
Does 4pi provide an EDS system for IBM-compatible PCs?(updated 10/26/97) Not as a complete solution; however, the SEII PCI card will work unmodified in a PC-compatible computer. 4pi supplies the necessary DLLs and drivers for using the SEII in a Wintel box, which will allow anyone to write their own PC-based EDS application for use with the SEII. 4pi can make the SEII PCI card available on an OEM basis for incorporation into PC-based systems. Please contact sales for more information.
I don't know about the status of higher level software: I've been using NIH-Image and XlispStat on the Mac. XlispStat is portable to many systems but would require Windows specific DLL's instead of the Mac ones to interface to the board. There is a PC version of NIH-Image, but I'm sure it also needs a different set of PC Plug-in's. ( I'm considering using Java and Image/J for the next iteration of this system, but even this would probably require some specialized native code. )
( But if you just get a Mac, you can use that and DTSA and FLAME and Photoshop and ... ;-)
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |--- All power corrupts and obsolete power corrupts obsoletely." - Ted Nelson
Dear Bruce Brinson: In regard to variability in partially saturated crystal images, I've received excellent info and recommendations from the people at Kimball Physics, Inc. (603-878-1616). We are running a style 90-15 crystal in our Philips CM-12 TEM. The first crystal lasted 3000hrs, the second about 1000hrs, and so far current one about 1000hrs. Although usually initial tip image was a classic maltese cross, it rarely remained so. After several hundred hours we usually see some tip "walking" which requires pulling the cathode and mechanically recentering the crystal. We always keep beam current low, 3 to 4 microamps at 120kv and bias 1 to enhance crystal life. We find that small black lines in the tip image can be removed by adding beam current but oversaturation may result. We always saturate using the light meter after making sure the image is centered electronically and the meter is of course a good way to monitor intensity loss over long periods of time. If we need a lower KV to enhance contrast we still try to keep filament current under 10 microamps although I think oversaturation is more likely to reduce crystal life. As to the static electricity problem, we work in a 10% RH environment because of cryo sample transfers. We leave the negs in metal holders until ready for transfer into plastic racks for processing. We used to obtain discharges as the films were dropped into the plastic racks until we started to prewet the racks. Sometimes the fireworks were a welcome distraction from the constant scratching! Don Gantz Boston Univ School of Medicine gantz-at-med-biophd.bu.edu
Jo, there is a company called Lamp Technologies Inc with a web page and on-line catalog at http://www.lamptech.com/info.html . The company supplies all kinds of lamps and has a special section on Microscope Lamps.
Joe Fu wrote:
} Does anyone still have the supplier list for the light bulb that used for } the optical microscope? TIA.
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
Over 10 years ago, in another lab, I ordered and received rubber rings designed to stretch-fit onto the outer rim of the lid of the round Pyrex glass vacuum desiccators. A lid could then be placed onto its base with the rubber ring establishing the seal, allowing one to forgo the use of vacuum grease and better protecting both components from damage. Because I used few of my desiccators to actually pull a vacuum, the seal was perfectly adequate. Are these still available? (Was it Denton Vacuum? Tousimis?) Surely someone will know and perhaps other labs besides mine will benefit.
Thank You
Scott Robinson EM Lab., UMKC Dental School, 650 E. 25th St. Kansas City, MO 64108 tel. 816-235-2072 robinsos-at-smtpgate.ssb.umkc.edu
A friend asked if I might know of a method to detect the identity of small contaminants (about one tenth to three tenths of a micron) on the surface of a photoresist coating on silica wafers. They have no good clues as to what it is, though it would help to know if it is organic. I have no experience with materials microscopy, though with a background in biological EM I wondered if OsO4 might be useful, since bound OsO4 could be detected via EM microanalysis. Perhaps there is a fluorescent dye which binds generic organics? If anyone has a suggestion, I would be very happy to take notes.
Many thanks,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
Dear Jim, William and list, I operate a TN5500 with a TN micro ZII detector. Could I ask you to keep me copied with the optioned offered to you? Thanks Ron Oleka -----Original Message-----
If the seed can be inbibed for a day or so fixation should easy, but of course this will cause physiological and structural changes. For dry seeds I would forget about glutaraldehyde fixation and use OsO4 at 30 degrees C. Try 15, 30 and 60 minutes. Be careful with the OsO4 fumes. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au } -----------------------------------------------------------------------. } } Can anybody tell us, what is the best way to fix and embedd tabacco seeds } for transmission electron microscopy? } We tried 2-times without success. One of these was by using seeds which } were pierced with a needle to get better penetration. We used relative long } times for dehydration/embedding. } Probably seeds were not fixed and/or lipid not penetrated by the embedding } medium. The lipid came out during sectioning. } Do we have to cut them completely to get penetration? If yes, how to avoid } the swelling and thus disturbing of parts of the seeds? Special procedures? } Is Epon better because of the lipids? } I have no experience with seed embeddings - unfortunately. } } Arthur } Dr. Arthur Schuessler } University of Heidelberg } Zellenlehre } Im Neuenheimer Feld 230 } D-69120 Heidelberg } Germany } } Fax: 06221 544913 }
Scoot: Cannot help with your rubber ring problem specifically, however, I note that you and most users of desiccators do not require a vacuum but just a dry atmosphere. Desiccating cabinets, especially the automatic ones which regenerate the desiccant used are the answer and a much better, maintenance free solution to dry storage than the quaint desiccators.
Disclaimer: ProSciTech supplies a large range of desiccating cabinets. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} -----------------------------------------------------------------------. } } Over 10 years ago, in another lab, I ordered and received rubber rings } designed to stretch-fit onto the outer rim of the lid of the round } Pyrex glass vacuum desiccators. A lid could then be placed onto its } base with the rubber ring establishing the seal, allowing one to forgo } the use of vacuum grease and better protecting both components from } damage. Because I used few of my desiccators to actually pull a } vacuum, the seal was perfectly adequate. Are these still available? } (Was it Denton Vacuum? Tousimis?) Surely someone will know and } perhaps other labs besides mine will benefit. } } Thank You } } Scott Robinson } EM Lab., UMKC Dental School, 650 E. 25th St. } Kansas City, MO 64108 tel. 816-235-2072 } robinsos-at-smtpgate.ssb.umkc.edu } }
Having just read the most recent "Net Notes" in Microscopy and = Microanalysis...could someone explain what a printer "enhancer" is?? Can = I use it in conjunction with a pre-existing network card??
We have an HP LaserJet IV series and we use it as a network printer. It = would be great if I could "beef" it up to reduce our Polaroid budget.
Nan H. Laudenslager Research Testing Group Specialty Minerals, Inc. Easton PA 18042
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Hello my name is Brandon Hernandez, I am currently a student at SJ Delta college's electron microscopy program. I was wondering if anyone has any information about FIB techniques, and theory. Is there a book? Any free information? Please contact me at brandon-at-gotnet.net Thank You!
Hello my name is Brandon Hernandez, I am currently a student at SJ Delta college's electron microscopy program. I was wondering if anyone has any information about FIB techniques, and theory. Is there a book? Any free information? Please contact me at brandon-at-gotnet.net Thank You!
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Hi Everyone, DTSA is Desktop Spectra Analyzer which is a Mac program developed by = Chuck Fiori that is good for teaching EDS purposes but does not have = several examples of multielement spectra in it for teaching purposes. It = does, of course, have pure element standards.
I need to have sample spectra so the students can practice the various = routines. We have a Kevex 8000 and Noran 5500 both of which are not = transferable to Mac systems (without adding an expensive board into the = system).
If there is someone out there in cyberspace that could send me spectra = that can go into DTSA, we would appreciate it. Thank You in advance for any help.
Judy Murphy Microscopy Technology Center San Joaquin Delta College Stockton, CA 209/954-5284
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This message is in MIME format. Since your mail reader does not understand this format, some or all of this message may not be legible.
I am looking to obtain a used SEM preferrably with EDS. The instrument will mainly used for metallurgical failure analysis. If you know of any instrument available, please contact me.
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{/HEAD} {BODY} {P} {FONT SIZE=3D2 FACE=3D"Arial"} I am looking to obtain a used SEM = preferrably with EDS. The instrument will mainly used for metallurgical = failure analysis. If you know of any instrument available, please = contact me. {/FONT} {/P} {BR} {P} {FONT SIZE=3D2 FACE=3D"Arial"} Ki Ho Lee {/FONT} {BR} {FONT SIZE=3D2 FACE=3D"Arial"} Lab Manager {/FONT} {BR} {FONT SIZE=3D2 FACE=3D"Arial"} Dexter Axle Division {/FONT} {/P}
} Date: Fri, 30 Jan 1998 09:53:55 } To: "Brandon j Hernandez" {brandon-at-gotnet.net} (by way of Nestor J. Zaluzec) } From: "Augusto A. Morrone" {amorr-at-mse.ufl.edu} } Subject: Re: FIB Info.? } } Brandon: } } You may start looking up several papers on FIB sample preparation in "Specimen Preparation for Transmission Electron Microscopy of Materials", MRS Symposium Proceedings,Vol 480, Edited by Ron Anderson and Scott Walck (1997). This proceedings has several papers on the technique (Gianuzzi et al, Phaneuf et al, Su et al, Shaapur et al, Tsujimoto et al) describing its application to various materials/site specific/XTEM/etc. } } Augusto } } At 02:24 AM 1/30/98 -0600, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
I apologize for scrambling my previous note to the list server. I actually sent ascii text, no pictures, etc. and have no idea what went wrong. If this message is scrambled, I will troubleshoot with our network police (who have absolute control and change things whimsically with no notice or accountability).
I only hope optical retardation is not inducing in me the mental kind.
My original question is below--please let me know if this is scrambled.
I want to observe interference colors using polarized light microscope and capture the image using a video camera. Using ImageTool, I can obtain RGB values of the colors at various points of the image. Using the RGB values only, I want to estimate the retardation. At the least, from the RGB values I should be able to "calculate" a color on a Michel-Levy chart to get a retardation. I realize I may not know which order I am looking at, but I may know other details about the sample to allow me to predict which order. For instance, based on RGB values only, I may be able to know the color is red and it would have associated retardation possibilities of 550nm, 1100nm, ... I would then "pick" the correct value based on my knowlege of thickness, typical birefringence values for the material, etc.
Question: can anyone image a conversion from RGB values to retardation values?
A useful program for generating crystal forms is Shape. Input the lattice parameters, space group, forms wanted and distance from the center and the program generates a crystal for you. Even in vrml. You can get a demo at
http://www.tricon.net/comm/shape/
There are versions for dos, windows 3.1, windows 95 and Mac. BTW the same old mac version that I ran on System 6.0 and a SE30 still runs on a PowerMac and MacOS 8.1.
You can get a demo for Atoms as well at the same site. This generates the crystal structure.
Cheers, Gordon
Gordon L. Nord Jr. Scientist Emeritus, Eastern Energy Team 956 National Center Office: 703-648-6745 U. S. Geological Survey FAX: 703-648-6419 Reston, VA 20192 gnord-at-mactem.er.usgs.gov USA http://mactem.er.usgs.gov/
I would like to thank all who answered my question concerning the separation of resin from glass. Thanks for the tips! However, I have a new question concerning the same matter. Is there any method for dissolving the glass from the resin? For instance, are there any known chemicals/solvents for this? The situation at hand is that resin stubs are stuck to microscope slides. Thanks for any assistance!!!!
James S. Mulroy Dept. of Neurology Emory University
A colleague of mine is looking for a means to fluorescently label bacteria prior to seeding onto a surface for testing. The dye must not interfere with normal functioning of the bacteria including adherence and growth. The result would be viewed under confocal or regular fluor. LM. Does such a label exist?
Acridine Orange was tried a while ago for a slightly different experiment and found to show too much bleeding under confocal. Also, UV excitation is out. On the Confocal Listserve there was a discussion some time ago mentioning dyes from Molecular Probes such as RH414, the Syto dyes esp. #10, Live/Dead, TMRE, as well as others. I have no personal experience with any of these. Does one stand out for this purpose?
Thanks as always, Karen
-- Karen Zaruba, kszaruba-at-mmm.com Life Sciences Sector Laboratory, 3M Center Bldg. 270-1S-01 3M Company, St. Paul, MN 55144
"If you can spend a perfectly useless afternoon in a perfectly useless manner, you have learned how to live." - Lin Yu Tang [Of course, a perfectly useless afternoon is a rare thing indeed!] *The opinions above are my own, not necessarily my employer's*
Dear all Zeiss LSM510 Users, Florence Niedergang asked about saving files and database, and copying them to MO disks. The easiest way to do this without losing any information or images when moving or coping files and databases is to Set Options of Saving. To do this go to Options menu in the LSM510 program. Click on Save page, and select the 3rd choice. This is "At Create Database" automatically create a subdirectory with the same name and write image files and database to that directory. This will create a directory in which all ones LSM files and the database.mdb files will be written. Then when one wants to move that to any other drive or media, or file server, you simple copy or move the entire directory. One can directly write to the MO drive as well, as it is not necessary to write directly to the hard disk. Simple choose that drive and a directory on it when you first create the new database. ( It allows you to designate where you want to do this). For multiple user instrument, users are able to have their own MO disks. I would not recommend using 230Mo's, as they are too small. I even find that the 640's are not large enough for some of the work we do. In regard to exporting images from these, users have a variety of file formats to choose from, tiff, Photoshop, JPEG, EPS, and GIF, etc. (Oh, by the way Zeiss, the GIF export currently does not function). I and my users export these to the users main directory, and after downloading to another location, delete them. All original databases and their LSM images are saved for archival, which can be returned to at any time in the future. To get the image or session information, about how one collected the images, one would down load a copy of the database file (.mdb) alone.
Joe Goodhouse Confocal Core Facility Molecular Biology Princeton University jgoodhouse-at-molecular.princeton.edu
To all MIK/MAS Members , CSMS members and Interested persons:
======================================= THE MIK / MAS --CSMS Spring 1998 meeting College of Veterinary Medicine University of Illinois Urbana , IL May 28-29, 1998 =======================================
NEW DATES:
NOTE CHANGE TO: May 28th & 29th 1998
These 2 societies will be having a joint meeting.
* Thursday May 28th will be a biological topics day. This will include our guest speaker Dr. Neville Cheville (Book: Cell Pathology) from Iowa State University.
**Friday May 29th will be a material Science topic day, including a tour of the Material Research Lab at the University of Illinois
**Thursday Evening will include a private show at our local planetarium.
Both Days will have mini workshops on :
-- Basics of setting up a Web Page on the Internet -- HANDS ON -- Digital Equipment usage. ___________________________________________
PRESENTATIONS:
We accept a large span of microscopy topics. Feel free to tell us what you would like to present.
If you have a presentation you would like to do, please contact me ( see info at bottom)
STUDENT COMPETITION:
Students are encouraged to present papers of work they have done in some microscopic area. Each day will have a $50 1st place winner. All student presenters also get a free lunch.
*********************************************************** Lou Ann Miller Center for Microscopy & Imaging College of Veterinary Medicine Dept. of Veterinary Biosciences University of Illinois Rm 1108 Basic Sciences Bld 2001 S Lincoln Ave. Urbana, Illinois 61802
Phone: 217-244-1566 email: lamiller-at-uiuc.edu
Center for Microscopy & Imaging Home page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society: http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/CSMS/csms.html
Personal Home Pages: http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/LAM.html
I passed your question on to Wesley Nieven at Surface Science Labs and he=
offered the following response. I hope it helps!
Best regards-
David Henriks South Bay Technology, Inc.
=46rom Wesley Nieven:
Contaminants on photoresist can be difficult especially if they are very thin, e.g. less than 0.5 micron. It is very doubtful at the 0.2 micron realm that histological or optical microscopy methods will work. There a= re several methods available, each giving different degrees/content of information about contaminant. =
1. The 'simplest' method is FTIR. However, 'simple' is not perhaps the best choice of words. Depending on contaminant film thickness, FTIR with=
ATR multi-reflection, you may be able to "see" the film. The photoresist=
background will need to be dealt with (a non-trivial matter). FTIR would=
require a substantial lateral size of the contaminant, say 100 micron or more for this technique to work. The FTIR would not identify a specific organic per se and would not identify a biological.
2. ESCA or XPS are very good at analyzing very thin films. Depth of information for XPS is about 100 Angstroms (0.01 microns). Lateral information area is about 10 microns. It can detect all elements greater=
than He at concentrations about 0.1-1.0% and give quantitative results (accuracy depends on standards, etc.) ESCA/XPS can also give some chemic= al state information, e.g. nitrogen as azide vs nitride, carbon as CFx vs carbide, etc. This can be extremely useful but chemical state info is no= t completely unambiguous. ESCA/XPS requires ultra-high vacuum ( {10e-9 torr= ) and samples must be compatible. Photoresist should not be a problem, but=
if contaminant has high volatility or is very hydrated (bio-mass) then th= is method may not work as is.
3. TOF SIMS (Time-of-Flight Secondary Ion Mass Spectroscopy) is a mass sp= ec method with extreme surface sensitivity. TOF information comes from top 2-3 monolayers of sample and can easily see films of one monolayer/monoat= om thickness. Mass resolution is good (typ. M/deltaM =3D ~10,000) and spati= al (lateral) resolution is reasonable (about 0.2 microns) but not simultaneously. TOF also requires ultra-high vacuum but a cold stage (offered by one or two of the TOF manufacturers) can work with volatile a= nd somewhat hydrated samples. Specific identification 'MAY' be possible wit= h TOF. Info from TOF is molecular/elemental mass fragments from surface. Complex organics can often be identified (with standard) and "reverse assembly" of molecular mass fragments can sometimes be done to yield exac= t parent molecule. TOF has excellent elemental/molecular sensitivity with some elements detected at ppb range or lower.
Caveats; the FTIR method, although more commonly available, is the least likely to work especially if the film is very thin and in small spots. FTIR for this application requires a very skilled analyst (your routine l= ab guy is not likely to be successful even if there is enough material to do=
the job) and a high quality machine (FTIR w IR microscope, multi-pass ATR=
cell, etc.)
ESCA/XPS is an expensive technique (instruments usually cost about $0.4 million) and requires experienced operator. Commercial analytical laboratories are probably the best bet. Cost for this analysis would probably run from $450-$1500 depending on what you need from the analysis= =2E TOF instruments are even more expensive (typ. $0.7million) and require ve= ry skilled, experienced analysts. There are not very many of these machines=
around the country. Commercial analytical labs are your only real choice=
here. Analysis would probably run around $750-$1000. =
Contact me directly and I can supply you with additional information. (a= nd also a commercial analytical lab as I am the Technical Advisor for one of=
them! {grin} We do this sort of analyses all the time.)
This note is a request for information on what you charge your users for SEM and TEM time and your opinion in determining if what we charge in reasonable.
On Wed., Feb. 11, 1998, I will be meeting with the folks who help me set rates for instrument use in my lab. They have asked me to come with some ideas about what other similarly equipped labs charge, part of an effort to establish a reasonableness factor in their calculations.
Here is our situation. This is a campus of abut 10K students with maybe 2K graduate students. We have no professional schools and I run the only EM lab on campus. We have a 1975 JEOL 100B TEM and a 1987 ISI WB-6 SEM. Both of these instruments function reasonably well and for the most part meet the needs of our users. We maintain service contracts on both instruments using an independent service provider at about $5K for each microscope.
For now we are charging everyone for supplies marked up (25%) to cover overhead like my time ordering. We charge funded researchers $10/hour (filament timer on TEM, best estimate for SEM) to use the microscopes. We do not charge students who are enrolled in a course that uses EM as part of the course. We don't make much money and so far that has been OK.
Most of our users are students from labs that do not have much grant support for EM. Some are undergraduates who may receive up to $75/quarter to support their independent studies courses, including EM if they use it. I would guess that our annual TEM use is between 250 and 500 hours and SEM use maybe twice that, some years a lot less.
As I go into this meeting I want to maintain low rates so the lab gets used, but I don't want to make it look like we give away everything unreasonably. If I were to calculate rates based on full cost recovery, the price would be out of reach for most of our users, or maybe not, I'm not sure. If we set the price too high, I think we would see use of the lab drop off.
What do you charge for your instruments and lab use? What sort of equipment do you have? What sort of campus are you on? Do you have different rates for different users? Are some users subsidized, and if so how? Given the vintage of our equipment and funding for our students, what would be a reasonable rate for us?
I know there are facility directories that have some of this information, but I was interested in some first hand data, even some of the subjective remarks that don't come through in a simple list.
Thanks, if you can help it will be great.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Scanners Corporation offers EDS upgrades for any manufactured system and {bold} NEW {/bold} EDS Analyzers, with either a premium 130ev detector or standard 140ev detector(both thin window). We also offer PC based solutions for SEM Digital Imaging, and of course SEM service contracts. For further information, please visit our website at www.scannerscorp.com or send me an email at gary.easton-at-scannerscorp.com (the period between my first and last name is important). Gary M. Easton, Pres. SCANNERS CORPORATION 5320 Enterprise Street Suite C Eldersburg, MD 21784 Phone 800-466-SCAN(7226) Fax 410-549-7583
First let me sincerely thank all (20-30) who responded to my query & now a summation of the responses: Most responses centered around the use the precautions one would use for C-MOS electronics. All indicated improvement but I am not sure any indicated complete success. The reason I did not pursue this approach is that unlike semiconductor devices which have nice leads for surface charges to traverse the device by, the charge on the film by my observation is trapped in the interior area between the sheets. Until an air gap develops, there is no way out. Another common denominator was the relative humidity, winter being worse. Some suggested wetting the area around anti-stat pads & wetting film trays. Though I've not had the experience, some told of discharging from film to film tray. It was also pointed out that peeling sheets apart rather than sliding the was advantageous & I agree but again not a complete solution (my experience). In general frictional movement, synthetic clothing & grooming your hair are on the not good list.. There was one interpretation, perhaps 2 noting that the film is driest just after coming out of the good TEM vacuum. I think this is a very valid point. The source of evil. Their experience was that if they returned the exposed film to the dissector over night (lower grade vacuum) that an improvement in ESD was noted. One step further might be to store it in the darkroom (ATM) a while. Humm may give that a go when anxiety ceases. Maintaining separation between film sheets with sheets of paper was also mentioned. Also opposite opinions, that film should be processed asap. Before the responses began arriving (really it's true), JD Wise & I chatted a bit. We came up with a solution which coincidentally is an application of several suggestions Bare in mind I have the constraints that I need to process asap, typically 50+ frames, in a 2nd dark room some distance from the TEM & working late at night I would prefer to reload the film magazine & leave it with the TEM.
My conclusion is that the charging originates when the film is removed from the film plates & stacked. From this point if the capacitor will charge, it probably has.
OUR CURRENT SOLUTION is base on this presumption, is a book out of the many anti electrostatic bags that lay about. Not envelopes, just sheets. Each sheet will be separated by an conductive spacer on the bound side. (one sheet of film thick to prevent wedging. Another option would be to spiral bind the sheets at the copy shop. The book will go into a box to keep it in order during transport. May add an antistatic mat to open the book on or just ground the binding. Once in the dark room I'll allow the sheets to slide part way out of the book while loading the film trays. I think the book approach will be more convenient that loose separators or many envelopes. It oughta be cheap, easy to make & prevent the build up of charge while maintaining a tidy package & be relatively easy to handle in the dark.. Enough FAB ......feed back?
disclaimer: I claim all copywriter, prior art, etc. & license anyone with a pulse to use this idea free of charge, well ok, the the dead & undead can use it too. Anyone selling it ought to go home & slap their daddy (after they send me 10%.)
The jury is still out on my filament question, in it's continuing evolution the cross is back with a sharp but off axis center bit. I've instructed our user group to record their best filament image at each session. Several responses indicated similar experiences but that the filament kept on trucking. Fickle behavior in the nominal 300 hour range was common & some were replaced others saw 1000-3000 hours of service. This problem may not be uncommon to the sharpened rod design. Most respondents did not have the Hi-res. applications & thus other than a different picture, their performance was not hindered while they had sufficient beam current. This may turn out to be a lot of filament travel, XZY. Eecchhhhh!
FYI the general opinion is that my Kimball filament is in the throughs of death at an early age. But when it was good, it was real good. Humm wonder if Kimball will stand behind it? (Kimball if ya want to contact me...713 285 5485.) Frankly the Denka before it was also short lived (~200hr) but then the 2000hr Denka before that didn't fail we just figured it would & swapped it. I went with a the KP not on price but based on what I considered a very solid recommendation from a JEOL user with years of KP exp. so I'm going to get another.
only a couple addressed the LaB6 cross. There was agreement that the cross originates from the crystal orientation (001), not the bulk geometry. I'll go with that.
I would like to submit the following abstract for an oral presentation. I believe it represents the introduction of an entirely new form of analytical microscopy that will soon be available as a commercial product.
Ive pasted the text and also attached a word7 document.
Sincerely
Dr. M Reading
Thermal Analysis For The 21st Century - mTA M. Reading, D. J. Hourston and M. Song, IPTME, Loughborough University, Loughborough LE11 3TU, UK. H. M. Pollock and A Hammiche, School of Physics and Chemistry, Lancaster University, Lancaster LA1 4YB, UK. T Lever, TAI Instruments, Leatherhead, Surrey UK, J Lecenby, 18 Hill Street, Essex CB10 1JD, UK. There are three major problems with our current thermal methods: the first is a purely practical one, experiments often take too long, especially thermomechanical measurements, the second is related to sampling. Frequently the sample is either too small, or too thin or buried within a larger component from which it is difficult to extract. The third is more fundamental, the information they provide is not spatially resolved. Atomic Force Microscopy (AFM) is a technique in which the tip of a probe is rastered over a surface to build up an image of the surface topography. Our apparatus is based on a conventional AFM where the tip of the probe has been replaced by an ultra-miniature resistive heater. The resistance also serves as means of measuring temperature, thus the tip, when used in conjunction with a reference probe, serves as a micro Modulated Temperature DSC cell. The tip is held at a constant average temperature and rastered over the sample surface in contact mode to build up an image. The data collected are the topography, as in traditional AFM, plus thermal conductivity, measured from the average (DC) signal plus thermal diffusivity, as measured from the response to the modulation (AC signal). Having imaged the sample, any point in the image can be selected and the probe tip is placed on it. The temperature of the tip can then be scanned in exactly the same way as conventional thermal analysis to obtain calorimetric measurements of transitions. In addition when the tip is placed on a selected point a carefully controlled force is applied to it. As the temperature increases the sample often softens and the probe indents further into the sample. This measurement is closely analogous to a ThermoMechanical Analysis (TMA) measurement. Both the calorimetric and mechanical property measurements are made simultaneously at heating rates in excess of 500 Celsius/minute. These micro-thermal analysis measurements solve all of the problems outlined in the opening paragraph. It also opens up a new range of applications for thermal methods in polymer science, catalysis, pharmaceuticals and composites by providing a powerful new form of analytical microscopy.
Douglas R. Keene asked the following: ============================================= A friend asked if I might know of a method to detect the identity of small contaminants (about one tenth to three tenths of a micron) on the surface of a photoresist coating on silica wafers. They have no good clues as to what it is, though it would help to know if it is organic. I have no experience with materials microscopy, though with a background in biological EM I wondered if OsO4 might be useful, since bound OsO4 could be detected via EM microanalysis. Perhaps there is a fluorescent dye which binds generic organics? If anyone has a suggestion, I would be very happy to take notes. =============================================== This is never a trivial kind of investigation. We have had this kind of problem, some times on polymer films or molded plastic parts and a few times on photoresist over the years. I am talking about features that are well enough into the submicron range in size that most of the other kinds of approaches that might come to mind just would not apply.
We have found that plain ordinary thin section TEM has often times provided either all the information needed to answer the question or else has made major strides in getting to the point where the question could be answered. And another advantage relative to the alternatives is that the TEM work, if managed properly, can be done for a fraction of the cost of the alternative approaches.
In the case of a polymer film, we first lightly coat with gold (sputtering) the film surface, then embed. The gold layer acts as a passivation layer to keep the embedding resin from dissolving or swelling or other wise interacting with the unknown particles. Usually, on the basis of electron contrast alone, one can make an educated guess as to whether the observed features are organic or something else and if the latter, EDS with SAED can provide even more information.
If on the surface of photoresist, the top surface has to be stripped off. After gold coating, we use as a stripping agent polyacrylic acid (PAA), and this is where we must keep our fingers crossed: We need at least some amount of the photoresist to strip off which sometimes happens and sometimes does not. Once stripped off, we lightly coat with aluminum that stripped surface (so the two surfaces are not confused once in section form and in the TEM), embed only that side, then in water dissolve away the PAA and embed that side as a secondary step. Now it is in a form to diamond knife thin section and the discussion would be the same as for the polymer film.
If the nature of the system was such that we could not get the photoresist to strip off with the PAA, we would then have to thin down in some way, the silicon wafer, probably first by mechanical milling and then a final step of plasma etching in a plasma etcher using CF4 gas. Then what is left can be embedded and thin sectioned.
Hope this information will in some way be of value.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We have purchased the VIDX X-ray microanalysis system. It is easy to use, inexpensive, and has overall great value. Latter this year, as money gets freed we will be adding active imaging and elemental mapping to the system.
Medjet Inc., a publicly held research and development firm specializing in the design and development of ophthalmic surgical devices, has a position available immediately within its Department of Clinical Studies. Preferred candidates will have completed a bachelors degree in the biological sciences with prior experience in all phases of light, scanning and transmission electron microscopy, photography, and animal handling. The individual will assist in the coordination, execution, and documentation of all research activities including clinical studies involving animal and human subjects. Job functions will include preparing specimens for histological analysis, operating and maintaining equipment within the histology facilities, and documenting the results of all analysis.
Headquarters are located in Edison, NJ. Interested applicants should forward a resume with cover letter to the attention of Daniel Caruso. Candidates will be considered until the position is filled.
Medjet Inc. Suite 301 1090 King Georges Post Road Edison, NJ 08837 Phone: (732) 738-3990 Fax: (732) 738-3984
I am responsible for organising an ESEM symposium in London, UK in July 1998 as part of Micro 98 under the banner of the RMS.I am soliciting contributed papers and/or posters for this meeting. There are a number of invited speakers presenting applications talks and I need the widest possible representation of users. If you wish to submit an abstract for consideration please contact either me or the RMS. Many thanks
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
In the past, the Epson Stylus Photo has received support as an inexpensive, near photo-quality printer. Has anyone had experience with the HP Photo Printer (~$100 more) and been able to compare the black and white and the color output of these two printers under meaningful conditions? In particular, is anyone using the HP printer with a Mac and, if so, how was the printer interfaced? In addition, any experiences with the negative/slide scanner (~$500) available from HP are also of interest. Thanks for any comments.
john
John Sutko Dept. Pharmacology/318 Univ. Nevada, Reno Reno, NV 89557
I am a student at NMSU and am studying developmental myelination on the TEM. Recently I've noticed that the myelin rings around the axons don not appear to show clearly defined intraperiod lines (only part of a line is visible and does not extend completely around...). Could this be due to fixation or dehydration/embedding? Are there any specialized protocols for myelin or protocols that limit the loss of lipids? I am using a standard biological protocol (dialdehydic fix, cacodylate buffers, post-fix osmium, dehydration-30,50,70,80,90,95,100-pproplyene oxide, embedding:araldite:EMbed (50/50), section, stain:uranyl acetae, lead citrate)
Do any of you have experience with analyzing residual packaging materials on small parts (i.e. electronic components, precision mechanical parts, samples submitted for F/A, etc.)? I'm primarily interested in success stories in which people have analyzed the components, found contaminants relating to packaging such as low-density polyethylene plastic, and then made a change to a non-contaminating packaging.
Thanks,
Harold J. Crossman Senior Scientist OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
Among other things, I run an EM service facility; I have 4 TEMs, 5 microtomes-3 with cryo attachments, and ancillary stuff. We don't do analytical analyses, but I can put you in touch with someone who does. We do biological TEM, thin sectioning, ultrathin cryosectioning for immunolabeling, negative staining, etc. Feel free to contact me if I can help you. See below.
On Tue, 13 Jan 1998, Anthony Domenicucci wrote:
} Date: Tue, 13 Jan 1998 09:12:41 -0500 } From: Anthony Domenicucci {domenicu-at-US.ibm.com} } To: Microscopy-at-sparc5.microscopy.com } Subject: Shared Facilities for STEM/TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am gathering information on Microscopy Facilities which rent time to } industry. I am interested in facilities which have the following capabilities } - e.g. High resolution TEM and STEM, EELS and Image Filtering, High Angle } Annular Dark Field. I would appreciate any help gathering this info. } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
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I am looking for a non-volatile, slightly viscous liquid with a refractiv= e index greater than 1.80. Are there are some new materials available w= ith these chracteristics?
by ursa.cus.cam.ac.uk with smtp (Exim 1.853 #1) id 0xzUtX-00046L-00; Mon, 2 Feb 1998 23:03:15 +0000
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-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.- Glynis de Silveira University of Cambridge Department of Materials Science and Metallurgy E-m:gds1002-at-cam.ac.uk Pembroke Street, UK Tel:+44(0)1223 334434 CB2 3QZ Fax:+44(0)1223 334567
I have an old (30++ yrs) Kinney that's going to salvage soon. The only number I can find on it is SC3CT. I do not have time to take it apart for you, but if you can use it and want to arrange to have it shipped, it's yours. Where are you?
On Tue, 20 Jan 1998, David L Johnson wrote:
} Date: Tue, 20 Jan 1998 08:23:59 -0600 } From: David L Johnson {jptmvl-at-mailbox.syr.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: high vacuum evaporator parts needed } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Community-- } We have a Kinney High Vacuum Evaporator (Model SC-2), and I'm trying to } locate an OFEC Hi Vacuum Globe Valve, size 1" (sweat type, straight } through). This is for the Backing circuit--the seat is OK, but the } brass bellows on the valve has given up. I contacted Kinney (now part of } Tuthill Corp) and they know nothing.... } jptmvl-at-mailbox.syr.edu } thanx } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} A colleague of mine is looking for a means to fluorescently label } bacteria prior to seeding onto a surface for testing. The dye must } not interfere with normal functioning of the bacteria including } adherence and growth. The result would be viewed under confocal or } regular fluor. LM. Does such a label exist?
I would suggest that you try the PkH2 dye from Sigma. We use this in mammalian cells, and are able to keep the cells alive for several weeks after labeling. The dye really screams. Of course, one would expect some decrease of intensity as cells proliferate, but this will be dependent on the number of doublings. One can always re-label cells as the proliferate. The dye is fixable as long as you do not use detergents or any extracting reagents, such as acetone and alcohols. It intercalates into lipid bilayers and fluoresces strongly with 488 excitation.
Joe Goodhouse Confocal Core Facility Molecular Biology Princeton University jgoodhouse-at-molecular.princeton.edu
We are looking for a high resolution color digital camera system as follows:
- hi res color digital camera (like the Leaf camera or better) - computer interfaced to the camera (prefer Macintosh but might consider Silicon Graphics) - lots of RAM in computer - CD writer (like APS Jaz/CDR combo) - large monitor, graphics tablet - image analysis package - ability to use camera on variety of light microscopes (Olympus, Leica, Nikon) - ability to use camera on copy stand
I invite vendors or satisfied users to send or phone me with comments & quotes. Please include $$ amounts. Of course, we need the info by Thursday of this week at the latest.
Many, many thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Dear Samantha, maybe my first reply was not sent correctly because of a electrical = circuit/net crash down in our hospital. Therefore once more again: maybe your problem is one of "sectioning plane". What about the = thickness of your ultrathins?; Do you have, and if yes, did you use and = prove ultrastructural appearance by means of a goniometer stage?
Hopefully it helps a bit, have a joyful day, best regards
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: S. CICERO[SMTP:scicero-at-NMSU.Edu] Gesendet: Montag, 02. Februar 1998 12:34 An: microscopy-at-sparc5.microscopy.com Betreff: TEM of myelin
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I am a student at NMSU and am studying developmental myelination on the TEM. Recently I've noticed that the myelin rings around the = axons don not appear to show clearly defined intraperiod lines (only part of = a=20 line is visible and does not extend completely around...). Could this be due to fixation or dehydration/embedding? Are there any specialized protocols for myelin or protocols that limit the loss of lipids? I am using a standard biological protocol (dialdehydic fix, cacodylate = buffers, post-fix osmium, dehydration-30,50,70,80,90,95,100-pproplyene oxide, embedding:araldite:EMbed (50/50), section, stain:uranyl acetae, lead citrate)
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I apologize for scrambling my previous note to the list server. I actually sent ascii text, no pictures, etc. and have no idea what went wrong. If this message is scrambled, I will troubleshoot with our network police (who have absolute control and change things whimsically with no notice or accountability).
I only hope optical retardation is not inducing in me the mental kind.
My original question is below--please let me know if this is scrambled.
I want to observe interference colors using polarized light microscope and capture the image using a video camera. Using ImageTool, I can obtain RGB values of the colors at various points of the image. Using the RGB values only, I want to estimate the retardation. At the least, from the RGB values I should be able to "calculate" a color on a Michel-Levy chart to get a retardation. I realize I may not know which order I am looking at, but I may know other details about the sample to allow me to predict which order. For instance, based on RGB values only, I may be able to know the color is red and it would have associated retardation possibilities of 550nm, 1100nm, ... I would then "pick" the correct value based on my knowlege of thickness, typical birefringence values for the material, etc.
Question: can anyone image a conversion from RGB values to retardation values?
My impression is that this is an artifact which is usually due to geometry rather than to fixation; if the myelin layers are not oriented precisely perpendicular to the plane of section, they will appear to be smeared in transmission. Since it is highly unlikely that this requirement will be met everywhere around an entire axon, you will see some areas which appear sharp and others which appear blurry. If you have a tilting stage, you may be able to test whether this is the case by tilting the sample and looking to see whether some fuzzy parts become sharp (this will depend on the angle of tilt of the plane of the myelin relative to the tilt axis of the sample holder).
Marie
} } I am a student at NMSU and am studying developmental myelination } on the TEM. Recently I've noticed that the myelin rings around the axons } don not appear to show clearly defined intraperiod lines (only part of a } line is visible and does not extend completely around...). Could this be } due to fixation or dehydration/embedding? Are there any specialized } protocols for myelin or protocols that limit the loss of lipids? I am } using a standard biological protocol (dialdehydic fix, cacodylate buffers, } post-fix osmium, dehydration-30,50,70,80,90,95,100-pproplyene oxide, } embedding:araldite:EMbed (50/50), section, stain:uranyl acetae, lead } citrate) } } Does anyone have any suggestions? } } Thank You! } } Samantha Cicero
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
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In the past, I have worked on getting a good fixation where I could visualize major and minor dense lines in the myelin. One post-fixation protocol that we came up with worked pretty well. It is at the Osmium post fix stage.
1% Osmium 1.5% Potassium Ferricyanide 0.1M Buffer (in your case, cacodylate)
Fix the same amount of time you normally would with standard Osmium.
Good luck,
Cheri Owen Neuroscience Imaging Core Emergency Medicine Wayne State University Detroit, MI 48201
We have set up a new light microscopy digital imaging station. Now we need to learn what to do with it!
Does anyone have feed back on good image analysis workshops that give a solid foundation in image analysis for the money? Or bad experiences? Or does anyone think it is better to just read the manual and muddle through?
Robert Underwood Morphology Core Univ. of Washington
Samantha, If you haven't gotten this bit of advice yet, go to potassium ferrocyanide reduced osmium, there is a refernce where it is used specifically for myelin preservation, (if you need that reference get back to me) it should do the trick.
Mike D JHMI Microscopy Facility
On Mon, 2 Feb 1998, S. CICERO wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } I am a student at NMSU and am studying developmental myelination } on the TEM. Recently I've noticed that the myelin rings around the axons } don not appear to show clearly defined intraperiod lines (only part of a } line is visible and does not extend completely around...). Could this be } due to fixation or dehydration/embedding? Are there any specialized } protocols for myelin or protocols that limit the loss of lipids? I am } using a standard biological protocol (dialdehydic fix, cacodylate buffers, } post-fix osmium, dehydration-30,50,70,80,90,95,100-pproplyene oxide, } embedding:araldite:EMbed (50/50), section, stain:uranyl acetae, lead } citrate) } } Does anyone have any suggestions? } } Thank You! } } Samantha Cicero } }
} ... In the TEM. Recently I've noticed that the myelin rings around the axons } do not appear to show clearly defined intraperiod lines (only part of a } line is visible and does not extend completely around...). Could this be } due to fixation or dehydration/embedding? Are there any specialized
Samantha:
If the line doesn't appear to extend completely around the myelin sheath, the most likely explanation is a slight tilt from perpendicular orientation in the section. Only textbook photos are always perpendicular or parallel to the plane of section. Try a tilt stage, and you may find that you can see the intraperiod line appear at one tilt angle and disappear at others. Because an axon and accompanying myelin are not perfect cylindars, one side may not be exactly parallel to the other side of the sheath. and thus both are not perpendicular to any one plane of section and only a partial intraperiod line will be seen.
If you still never see the expected layers, then I would start to look for other causes, such as fixation or chemical exposures of the animals before fixation.
-Dennis
Dr. M. Dennis Goode Phone (301) 405-6917 Department of Zoology Fax (301) 314-9358 University of Maryland e-mail goode-at-zool.umd.edu College Park MD 20742 ************************************************************* "If the Lord Almighty had consulted me before embarking upon the creation, I should have recommended something simpler." -Alphonso X of Castile, 15th Century
I am looking for suggestions on features we should consider for an EM lab renovation. We are currently in the design process for renovating a building which will house two TEMS, 2 SEMs, an AES, and a darkroom. With this opportunity to design the EM labs, we want to take all reasonable precautions and make the necessary improvements to optimize these areas. This includes necessities for EM operation as well as conveniences.
While this currently unoccupied building readily passes vibrational and magnetic field tests, I am trying to minimize the impact of the labs, offices, and electrical/ventilation systems which will surround the EM labs. Alderson's book (suggested on the list a while back) was a great help for the initial design stages. What suggestions do you have either for the design of the laboratories or for the related equipment? While my primary interest is for the TEM labs, I would welcome any suggestions for the other EM rooms or dark room as well.
One major concern I have is with mechanical vibration isolation. We would like to limit a priori the effects of the surrounding labs and services. Any suggestions which would limit the effects of mechanical vibrations at the TEMs, or reduce the level of ambient vibrations, would be particularly helpful.
Richard Fonda
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
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Topic: Microwaving for SEM We have been experimenting with the microwave for TEM, and want to use it = also for SEM samples, which of course are larger. We do both Glut, Os, = and conductive staining techniques.
Questions Does anyone have experience with microwaving for SEM samples? How large of samples do you use? What protocols do you use? and times in the microwave? Wattage and type of microwave? Do you use iced samples or any special conditions for SEM samples? Any thoughts on results as compared to standard preps?
Thanks for any info. We are setting up experiments and need some = starting point for SEM samples.
Thanks in advance for any input. Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
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Listers,
I know a lot of you are real smart when it comes to computer images & stuff. I have an optical disc drive on my SEM and would like to go to something that most people have (we have the only optical disc drive on campus). Folks here have suggested Zip Drives. Now, I'm an idiot when it comes to computer stuff, is a Zip drive good enough for storing images and will people get decent images back when they put them on their lab computers? I've heard that Zip drives and the discs are fairly inexpensive, is that true?
Thanks in advance for all your fine help.
Still preferring to make photographs,
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
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Dear Richard, } } One major concern I have is with mechanical vibration isolation. We would } like to limit a priori the effects of the surrounding labs and services. } Any suggestions which would limit the effects of mechanical vibrations at } the TEMs, or reduce the level of ambient vibrations, would be particularly } helpful. } You are right to be concerned. One difficulty is that there is no one prescription for minimizing mechanical vibrations. If the building and ground is very rigid, it will transmit vibrations due to traffic and wind, so you must isolate the equipment with, e.g., a pneumatic platform, but if the building and ground are not rigid, and do not transmit vibrations read- ily, you might need to anchor the equipment so that, e.g., air conditioning vibrations do not affect the EMs. Furthermore, the frequency spectrum of the vibrations is important. Our HVEM responds greatly to 20 Hz vibrations, but not to ~22 Hz vibrations. Sometimes a spring-mass-spring type of moun- ting--as we use for our vacuum pumps--will lower transmission of vibrations, and it can be tuned to damp specific frequencies. Good luck. Yours, Bill Tivol
} Folks here have suggested Zip Drives. Now, I'm an idiot when it } comes to computer stuff, is a Zip drive good enough for storing images
If the capacity is large enough to hold the image, then, yes. The important thing is the information--the string of 0's and 1's--not the form it's written in (assuming your equipment can read that form). Thus, the only necessary criterion is the capacity. There are also considerations of convenience, i.e., can one read in the info in a reasonably short time or is the info compatable with other interested parties' equipment. The Zip drive does well on both counts.
} and will people get decent images back when they put them on their lab } computers?
If and only if the images were decent in the first place. The digital info will be read without added noise [of course all computers are perfect, aren't they? Oh, right, the early pentiums. ;-)]. Zip disks should certainly hold the info without significant loss, so they will pass this test.
} I've heard that Zip drives and the discs are fairly inexpensive, is } that true? } Yes. } } Still preferring to make photographs, } They still record much more info than any available digital system. Yours, Bill Tivol
We are considering Image Pro Plus (IPP) from Media Cybernetics for automating image acquisition and analysis. Does anyone know if Image Pro commands can be driven from Visual Basic, such as through an Active X or similar control? The promotional literature I have seen so far for IPP only mentions VB in terms of within-program macro scripting. (No luck contacting Media Cybernetics or its local vendor yet). We are thinking of using Visual Basic to be the core of a comprehensive microscope automation, image acquisition, and image analysis program. Any other ideas or tips would be very appreciated. Thank you.
Cynthia J. Zeissler Physical Scientist National Institute of Standards and Technology cynthia.zeissler-at-nist.gov 301-975-3910
We've just had our EDS detector crystal replaced and I am now getting complaints that the copper peak that is present whenever we collect spectra (including hole counts) is too large. We've placed the repaired detector at the same position as before and are using a Pt top-hat aperture. My questions: 1) What is the source of the Cu peak when we collect a hole count? 2) How can I minimize the presence of the Cu peak? 3) How big of a Cu peak, is too big of a Cu peak?
} ..., } } I know a lot of you are real smart when it comes to computer images } & stuff. I have an optical disc drive on my SEM and would like to go to } something that most people have (we have the only optical disc drive on } campus). Folks here have suggested Zip Drives. Now, I'm an idiot when it } comes to computer stuff, is a Zip drive good enough for storing images and } will people get decent images back when they put them on their lab } computers? } I've heard that Zip drives and the discs are fairly inexpensive, is } that true? } } ...
One virtue of optical drives (... over zip or jaz drives ...) is file integrity and longevity ... that is, if you want to consider "archival" abilities, then stay with optical. In this regard, you probably want to only consider one possibility, that being a CD writer ... the prices are way down and the media is less than $10/600Mb ... you get archival quality optical and a CD compatible across all platforms. The only disadvantage to access times for writing and reading files ... CDs will not compete with magnetic media in this regard.
... hope this helps :o) cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility at the University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
We are using an Agfa Duoscan (1000 dpi) for digitizing HRTEM images.
At the moment we are saving the images to disk, transfering the tif files via Apple File Exchange to the Mac, then using Digital Micrograph to give us an FFT, for printing/exporting etc.
Question:
Is there software available, (freeware or...) that would allow us to perform the "FFT" on the "PC" that we are using to digitize the HRTEM?
Thanks in advance
Fred Pearson Electron Optics Coordinator
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
Salzburg, 3rd of Febr., 1998, local time 10.55 p.m.
Dear listmembers, I am confronted with a type of resin, called "LX 112", which obviousely = is not available via a European EM supplier or at least very unusual to = use in Europe=20 (if this is false, please correct me). Concerning questions for the POP-OFF-technique (previous postings) I = learned of the use of that resin "LX 112". It seems to me either to be a = substitute for EPON or a Spurr's like resin.
I know from Abstracts in the MSA-Proceedings that MASCORRO J.A. and = KIRBY G.S. described Novel EPOXY/Anhydride Alternatives.....(EMBED 812 & = LX112), MSA Proc. 47th Ann.Meeting, 1989, p.1000/1001, & MSA Proc. 49th = Ann Meeting, 1991, 292/293, &.....EM-VIEWS (Texas A&M Univ., Issue #8, = 1993, p. 23-24)
Unfortunately, no supplier address, physical properties, etc. were = mentioned (maybe it is an invention of J. A. MASCORRO himself?).
If anyone out there could provide me with the address data of a = supplying company, even in the USA, and some comments on working with it = (for instance, experience with hardener, catalyst and or accelerator, = embedding quality, physical properties like the type of polymerization, = etc) it would be a great help for me (and Eva KELLER: enjoy the day, = greetings!).
Thanking you in advance very best regards
Wolfgang
__________________________________
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L") _______________________________________________________________
The Microscopy and Microanalysis Center at the University of Maryland at = College Park is searching for an assistant to help maintain two TEMs, = one electron microprobe and an environmental SEM. The MMC is a campus = facility that provides service to faculty, students, and outside users. = The facility is also used for teaching and research. The qualified = candidate should have experience in the maintenance of electron = microscopes and their use. Background on electronics and vacuum = technology is required. The starting salary is $30,000 to $35,000 = depending on experience. =20
Interested candidates should send resume and list of three references = to:=20 Lourdes Salamanca-Riba at either riba-at-eng.umd.edu,=20 Fax No. (301) 314-9467, or=20 Materials and Nuclear Engineering Department University of Maryland College Park, MD 20742-2115
The University of Maryland is an equal opportunity affirmative action = employer.
} } Does anyone have feed back on good image analysis workshops that give a } solid foundation in image analysis for the money? Or bad experiences? } Or does anyone think it is better to just read the manual and muddle } through?
Obviously my reply is suspect since I teach the course, but over the last 15 years we've had more than 1000 students attend the three-day workshops on quantitative image analysis that we teach at N. C. State University every May, and most of them tell us they feel the course has been worthwhile, and prove it by sending their colleagues. Info is available on-line at http://members.aol.com/IPCourse
First, what kind of optical drive do you have? We have an HP magneto-optical which will store about 650 MB on each side of a disk (1300 MB total). But it too is the only one we have found in the area, and it is going bad. It will not reliably accept cartridges. Once it takes a cartridge, it appears to be fine. It would be great if you might be able to help us out in a pinch.
Now as to your question, ZIP drives cost $15 or less per 100 MB cartridge. That it is a little expensive and small. The good point is that they are becoming fairly common. Thus it may be a good media for passing around. We have one, but don't use it much yet. The internal IDE or SCSI variety will give much better performance than the external parallel variety. They also now have a combination model to work with SCSI or parallel. It auto-senses.
I might suggest a CD writer. The platters are fairly cheap - less that $3 per 600 MB disk. The writers are coming down in price and are available for under $500. The main benefit is just about everybody and their brother (or sister) have one readily available. We are using one from HP and it seems to be working fine for us.
Hope this helps some.
At 12:12 PM 2/3/98 -0800, you wrote: } Listers, } } I know a lot of you are real smart when it comes to computer images } & stuff. I have an optical disc drive on my SEM and would like to go to } something that most people have (we have the only optical disc drive on } campus). Folks here have suggested Zip Drives. Now, I'm an idiot when it } comes to computer stuff, is a Zip drive good enough for storing images and } will people get decent images back when they put them on their lab } computers? } I've heard that Zip drives and the discs are fairly inexpensive, is } that true? } } Thanks in advance for all your fine help. } } } Still preferring to make photographs, } } } Paula = ) ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
Zips are a good choice, the drives and disks are relatively inexpensive. You can install them on both Mac and PC platforms easily. You can also get some cheap software that allows you to read the Mac formatted drives on PC's. I use Conversions Plus, I buy PC disks, and quick format them in Mac format (takes 8 seconds). This has solved my long filename compatibility problems across the two platforms.
With respect to storage, garbage in garbage out. If you have good digital images, they will retain their quality, it is not a function of the storage media that you use. Copying a digital image many times does not degrade the quality of the image.
The number of images on a disk is dependent on the file size of the images and the bit resolution. I good rule of thumb for a grey level image with 8 bit level is that an image in the 1 Mbyte size range will give you a reasonably good image in the 4 x 5 inch size at a resolution of 300 dpi. you can put about 90 such images on the ZIP disk.
However, things get bigger with high resolution and number of color channels. If you collect a 10 or 12 bit image, it will still have to be stored with a16 bit format; the file size will be twice the size of the 8 bit image (8 bits of depth gives you the 256 gray levels). If your image is color and you choose RGB mode (i.e. 3 color channels) then you will need to multiply the size by 3. If you double the image resolution, e.g. change 300 to 600, you will need to multiply the file size by a factor of 4.
Example: a 1Mb file at 8 bit, grey scale, and 300 dpi would be 24Mb (1 x 2 x 3 x 4) for a color(x3), 12 bit (x2), 600 dpi (x4) image.
I hope this helps.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I know a lot of you are real smart when it comes to computer images & stuff. I have an optical disc drive on my SEM and would like to go to something that most people have (we have the only optical disc drive on campus). Folks here have suggested Zip Drives. Now, I'm an idiot when it comes to computer stuff, is a Zip drive good enough for storing images and will people get decent images back when they put them on their lab computers? I've heard that Zip drives and the discs are fairly inexpensive, is that true?
Thanks in advance for all your fine help.
Still preferring to make photographs,
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
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Reply to: RE} FFT programs for PC
Hi Fred,
we recently bought an Image Processing toolkit with plug-ins for Photoshop. It runs under Mac as well as Windows and has, among a lot of other things, FFT. The package is from Reindeer Games. I haven't tested it very severely but it did give me good power spectra. Unfortunately the documentation is rather limited and I can't find their full address.
Hope this helps anyway.
Nick Schryvers
--------------------------------------
We are using an Agfa Duoscan (1000 dpi) for digitizing HRTEM images.
At the moment we are saving the images to disk, transfering the tif files via Apple File Exchange to the Mac, then using Digital Micrograph to give us an FFT, for printing/exporting etc.
Question:
Is there software available, (freeware or...) that would allow us to perform the "FFT" on the "PC" that we are using to digitize the HRTEM?
Thanks in advance
Fred Pearson Electron Optics Coordinator
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
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Dear microscopist,
some weeks ago I asked for adresses for purchasing collodium. Thanks to all who answered me! Now I got pyroxilin (= collodium) 2% in amylacetate. When I put one drop onto water, it spreads incredibly. So the resulting film is much to thin and tears immediately when I get the grid into the EM. Is the 2% solution to weak? Is there another method to produce a thicker film?
I am trying the collodium support for I get enormous binding of the *primary* antibodies to the formvar sometimes. I find it often, that primary antibodies bind to formvar, but not to that extend that I saw with two antibodies from eggyolk and rabbit. Any suggestions how to overcome this?
Thanks a lot in advance
Birgit
Dr. Birgit Neubohn Institute of Plant Genetics and Crop Plant Research (IPK) Corrensstr. 3 D-06466 Gatersleben-Deutschland
Several years ago, we were in a similar situation. The company renovated a vacant building so all us R&D folk could play in the same place. Of course it passed inspection - there was nothing in it. After moving, we found that we ran into numerous problems with EM fields for a variety of reasons. We eventually found the sources and had the problems corrected, but with quite a bit of unnecessary expense and down time.
If I had to do it again, I'd bring in the vibration and field experts during the design stage.
If you'd like to, contact me at the address below and I can give you a reference.
Harold J. Crossman Senior Scientist OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com }
} Is there software available, (freeware or...) that would allow us to } perform the "FFT" on the "PC" that we are using to digitize the HRTEM?
Try simple and fast program (I belive it is freeware) ProFFT (pro stands for Project and not Professional). Pics should be in *.bmp format. The program can be found at:
ftp://ftp.cdrom.com/.5/asme/WIN_ENG/PROFFT.ZIP or ftp.wustl.edu/systems/ibmpc/umich.edu/windows/graphics/bmp/profft.zip or ftp.iij.ad.jp/win3/desktop/profft.zip or ...
Limitations: (from readme.txt): The pictures to be transformed has to have the following characteristics: They must be square and their width and height has to be in a power of 2 (16, 32, 64, ..., 2^n). They must be in the DIB (Device Independent Bitmap) format specified for Windows 3.X and OS/2. In addition they must have 8 bitplanes (that is a maximum of 256 colours/grayscales) and must have a grayscale palette. The palette is presumed to have colour 0 as black all the way up to 255 as white (so that 127 equals 50% gray).
To all, A client of ours is urgently in need of a condenser aperture holder for the Philips EM 400, early model. We only need the insert, not the whole assembly with the bellow.We would appreciate any help. Please call George at KAM CONSULTING at 718-729-1997or E-Mail him at: dimitri-at-interport.net or E-Mail me directly or call 215-699-6160 Thank you, Peter A. Stolzenberg, PESTO INC.
For doing immunoelectron microscopy I would suggest that if at all possible you completely eliminate the collodion and formvar films.
Also, the kind of metal that is used in the grid seems to somehow affect non- specific binding as well.
If it is a question of supporting your sections during the immunolabeling, I would suggest you use a fairly high mesh (300 or 400 mesh) gold grid, preferably with a hexagonal mesh pattern, with no supporting film. Just put the sections on the grids, dry them, and begin the immunolabeling procedure.
If this is not possible, and you absolutely must use a supporting film, you can try increasing the salt concentration in your buffer washing steps. If you are using phosphate buffered saline or any other recipe with NaCl in it, it is probably around 150 mM (0.9%). Try boosting the NaCl to 5X normal. This would make it 750 mM, or 4.5% by weight.
If you use the high salt buffer, remember to incubate the grids in a couple of changes of regular strength (150 mM) saline before you go to the next step, to get the salt back into the range of physiologic strength! High Salt concentrations will often get rid of non-specific binding.
I hope this is of some assistance. Let us know how it works.
Best regards, Bob ********************************* Robert (Bob) Chiovetti E. Licht Company / USA / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Leica (Wild, Leitz, Bausch&Lomb, Cambridge, AO, Reichert-Jung) / Technical Instrument Company / American Volpi / Fostek / Stocker and Yale / AEI North America / OptiQuip / Dolan-Jenner / Osram / G.E. / Philips / Ushio / Boeckler Instruments / Heidenhain / Narishige / Colorado Video / Visual Environments of California, Inc. / Kinetic Systems / Pacific Precision Laboratories, Inc. / Pryor Scientific / Compumotor / Sutter Instrument Co. / Advanced Database Systems / Cohu / Javeline Electronics / Optronics / Diagnostic Instruments, Inc. / Dage MTI / Hitachi / Panasonic / Polaroid / Kodak / Mitsubishi / Sony
Regarding John Russ' reply: I am a graduate of the NCSU course and readily recommend it to anyone who is looking for a solid foundation in digital imaging. I hope my response is not as "suspect" as John's -the only "kickback" I get from the course is the knowledge!
There is a software called Scion Image. It's a software based on NIH Image ( a Mac based freeware from NIH). You can download it from the web site( http://www.scioncorp.com) or call 301-695-7870 for help.
Best regards,
On Tue, 3 Feb 1998, Fred Pearson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } We are using an Agfa Duoscan (1000 dpi) for digitizing HRTEM images. } } At the moment we are saving the images to disk, transfering the tif files } via Apple File Exchange to the Mac, then using Digital Micrograph to give } us an FFT, for printing/exporting etc. } } Question: } } Is there software available, (freeware or...) that would allow us to } perform the "FFT" on the "PC" that we are using to digitize the HRTEM? } } Thanks in advance } } Fred Pearson } Electron Optics Coordinator } } ******************************************************** } Fred Pearson } Brockhouse Institute for Materials Research } McMaster University } 1280 Main St. West } Hamilton, Ontario } Canada L8S 4M1 } } ******************************************************** } }
Tseng-Ming Chou (Alex) Dept. of Materials Science and Engineering Stevens Institute of Technology Castle Point on Hudson, Hoboken, NJ 07030 e-mail: tchou-at-attila.stevens-tech.edu tchou-at-menger.eecs.stevens-tech.edu The Microstructure Group of Stevens
One idea is to get both a Zip Drive and a CD read/writer. Store your images on the Zip disks and when you get six to eight filled up, transfer the files to a CD. Then recycle the Zips. The Zip drive is more agile when working with individual files. Writing to the CD is best done in a few (6 to 8 if you are copying from 6 to 8 Zip disks) sessions. Using the Zip disks allows you to make changes before archiving the final file(s). Often a lab or department with several computers used in image handling will have Zip Drives on each computer, and a single CD writer that is external & portable. This arrangement will give you the write/read/change/write again flexibility of the Zips and the efficient & permanence of the CDs. And you won't have to buy truck loads of Zip disks......just a car load of CDs.
Joiner Cartwright, Jr., Ph.D. Assistant Professor of Pathology Baylor College of Medicine Houston, Texas U.S.A.
2% should be o.k., but collodine solutions have this wonderful property in that you can stack the layers. I usually use 4%, I gently place one drop on the watre surface, as you've noted as the drop spreads you note that it reaches maximum spread and then 'bounces" back a little, right when it bounces back I add one more drop to the middle. The second drop spreads on top of the first giving a thicker film. You can use the color index (just like sectioning) to tell you approximately how thick the film is and just keep adding drops of the 2% until you get a color/thickness you like.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
A couple of related notes on recent postings . . .
The first posting: } } Does anyone have feed back on good image analysis workshops that give a } } solid foundation in image analysis for the money? Or bad experiences? } } Or does anyone think it is better to just read the manual and muddle } } through? } } Obviously my reply is suspect since I teach the course, but over the last 15 } years we've had more than 1000 students attend the three-day workshops on } quantitative image analysis that we teach at N. C. State University every May, } and most of them tell us they feel the course has been worthwhile, and prove } it by sending their colleagues. Info is available on-line at } http://members.aol.com/IPCourse } } John Russ
And the second posting: } we recently bought an Image Processing toolkit with plug-ins for Photoshop. It } runs under Mac as well as Windows and has, among a lot of other things, FFT. } The package is from Reindeer Games. I haven't tested it very severely but it } did give me good power spectra. Unfortunately the documentation is rather } limited and I can't find their full address.
It turns out the the Image Processing Toolkit is a companion to John Russ's book "The Image Processing Handbook." They can be purchased, or received when attending the image analysis course at NC State. I've attended the course and feel it was well worth the money. Definitely better than "muddling through"!
The toolkit actually has pretty decent documentation in the form of "tutorials" in pdf (Adobe Acrobat, a reader for which can be downloaded free--from www.adobe.com I believe) format on the CD. I'm sure info on the toolkit can be found at the web address Dr. Russ gave above.
Disclaimer--I have no affiliation with Russ, NC State, or Reindeer Games Software. I'm just a satisifed customer!
Jim Passmore Analytical Chemist Cryovac North America
There is a lot of overhead (ie wasted capacity) involved in storing files=
to CDROM. For one at a time storage of images, the overhead will use a lo= t more disk than the image. Best thing is to batch the images on to Zip or hard drive, wait till you got a lot, hopefully at a logical break in the stream of images, then archive them to CDROM. Some labs I know buffer images on the SEM hard drive, push the images in batches up a network to a departmental server, then recycle the SEM stora= ge space. The deparmental server has a CDROM writer on it, when a suitable block of images (generally a full CDROMs worth), is collected, a CD is written, and the server space recycled.
Thank you to those who responded to my question about methods to detect contaminants on the surface of photoresist. As I mentioned in my message, I am a biological microscopist. I gained alot of respect for those of you doing materials microscopy...you even know a language that I have never heard before! The responses are listed below, for anyone interested.
My original message to the microscopy list server (microscopy-at-Sparc5.Microscopy.Com):
Hello to the Microscopy group!
A friend asked if I might know of a method to detect the identity of small contaminants (about one tenth to three tenths of a micron) on the surface of a photoresist coating on silica wafers. They have no good clues as to what it is, though it would help to know if it is organic. I have no experience with materials microscopy, though with a background in biological EM I wondered if OsO4 might be useful, since bound OsO4 could be detected via EM microanalysis. Perhaps there is a fluorescent dye which binds generic organics? If anyone has a suggestion, I would be very happy to take notes.
Many thanks,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
Dear Doug:
I passed your question on to Wesley Nieven at Surface Science Labs and he offered the following response. I hope it helps!
Best regards-
David Henriks South Bay Technology, Inc.
} From Wesley Nieven:
There are several methods one could use. They are highly dependent on analyst skill and experience. There is some degree of dependence on equipment capability, e.g. a cheap, 'routine' FTIR instrument would not do the job. Other methods that can "see" films/contaminants this small will not give info you can use, e.g. AFM would likely see the contaminant but would not provide any chemical or ID information (only topography).
Contaminants on photoresist can be difficult especially if they are very thin, e.g. less than 0.5 micron. It is very doubtful at the 0.2 micron realm that histological or optical microscopy methods will work. There are several methods available, each giving different degrees/content of information about contaminant.
1. The 'simplest' method is FTIR. However, 'simple' is not perhaps the best choice of words. Depending on contaminant film thickness, FTIR with ATR multi-reflection, you may be able to "see" the film. The photoresist background will need to be dealt with (a non-trivial matter). FTIR would require a substantial lateral size of the contaminant, say 100 micron or more for this technique to work. The FTIR would not identify a specific organic per se and would not identify a biological.
2. ESCA or XPS are very good at analyzing very thin films. Depth of information for XPS is about 100 Angstroms (0.01 microns). Lateral information area is about 10 microns. It can detect all elements greater than He at concentrations about 0.1-1.0% and give quantitative results (accuracy depends on standards, etc.) ESCA/XPS can also give some chemical state information, e.g. nitrogen as azide vs nitride, carbon as CFx vs carbide, etc. This can be extremely useful but chemical state info is not completely unambiguous. ESCA/XPS requires ultra-high vacuum ( {10e-9 torr) and samples must be compatible. Photoresist should not be a problem, but if contaminant has high volatility or is very hydrated (bio-mass) then this method may not work as is.
3. TOF SIMS (Time-of-Flight Secondary Ion Mass Spectroscopy) is a mass spec method with extreme surface sensitivity. TOF information comes from top 2-3 monolayers of sample and can easily see films of one monolayer/monoatom thickness. Mass resolution is good (typ. M/deltaM = ~10,000) and spatial (lateral) resolution is reasonable (about 0.2 microns) but not simultaneously. TOF also requires ultra-high vacuum but a cold stage (offered by one or two of the TOF manufacturers) can work with volatile and somewhat hydrated samples. Specific identification 'MAY' be possible with TOF. Info from TOF is molecular/elemental mass fragments from surface. Complex organics can often be identified (with standard) and "reverse assembly" of molecular mass fragments can sometimes be done to yield exact parent molecule. TOF has excellent elemental/molecular sensitivity with some elements detected at ppb range or lower.
Caveats; the FTIR method, although more commonly available, is the least likely to work especially if the film is very thin and in small spots. FTIR for this application requires a very skilled analyst (your routine lab guy is not likely to be successful even if there is enough material to do the job) and a high quality machine (FTIR w IR microscope, multi-pass ATR cell, etc.)
ESCA/XPS is an expensive technique (instruments usually cost about $0.4 million) and requires experienced operator. Commercial analytical laboratories are probably the best bet. Cost for this analysis would probably run from $450-$1500 depending on what you need from the analysis. TOF instruments are even more expensive (typ. $0.7million) and require very skilled, experienced analysts. There are not very many of these machines around the country. Commercial analytical labs are your only real choice here.
Analysis would probably run around $750-$1000.
Contact me directly and I can supply you with additional information. (and also a commercial analytical lab as I am the Technical Advisor for one of them! {grin} We do this sort of analyses all the time.)
Phone (650) 962 8767, 800 321 4775 Fax (650) 962 0923 e-mail: wnieveen-at-surface-science.com ------------------------- You can try FTIR using diamond anvil cell microscopy/FTIR technique. -------------------------- Perhaps you should find someone with some surface analytical equipment - x-ray photoelectron spectroscopy (will give elemental and chemical state information, analysis depth ~5 nm, area 50 um - 1 cm across depending on instrumentation), secondary ion mass spectrometry (especially time-of-flight SIMS) (will give fingerprint mass spectra, especially useful if you have your unknown and some candidates under suspicion and good for differentiating between different organics of similar structure, analysis diameter say .1 um upwards, analysing outer monolayer of contaminant, could etch through to guestimate thickness, could create maps to see if coverage uniform. Probably better in this case as carbon and oxygen spectra in XPS can be a pain to analyse. Another possibility, though the size/thickness of your contamination might be a problem here, could be Raman spectroscopy (very akin to infrared spectroscopy) if you can find one locally with microanalysis possibilities. Again (as with anything, really) if you have any candidate contaminants to compare the unknown to, it makes life easier.
Of course, none of this is useful if you either don't have access to the equipment or money to pay for time.
Silicon wafer samples are great for surface analytical machines (much the same as for microscopes I guess) - nice and flat, can easily be cut to size
Best of luck,
Keith
-- Dr. Keith R. Hallam University of Bristol, Interface Analysis Centre, Oldbury House, 121, St. Michael's Hill, Bristol, BS2 8BS, England Telephone: + 44 (0)117 925 5666 | E-mail: k.r.hallam-at-bristol.ac.uk Facsimile: + 44 (0)117 925 5646
| URL: http://zeus.bris.ac.uk/~phkrh/
Hi Doug,
We routinely use EDS either in a Hitachi S-4700 FE-SEM or an ADEM Lab6 SEM to detect trace contaminates on Photo Resist. Typically, the eV energy of the element that may be present times 1.5 will give you the desired beam energy to use. I would suggest starting at lower KeV and move upward. We also use a backscatter detector to image instead of the secondary detector due to charging.
Deposits on photoresist are a big problem in the micro circuit industry, finding out what they are may be equal in status!
Lets talk you through a SEM protocol.
1. Photoresist is made up of very light elements which will help in the SEM as other materials will almost certainly be heavier and be better imaged by the system, this will enable you to see detail down to tenths of a micron with no problems.
2. Your analytical problems are much greater. To display a reasonably good range of light elements in an EDX system we need at least 10kV. The problem is that for carbon we are punching the beam in at least 1.3 microns (by the Monte Carlo calculation) so to be certain that the only information comes from the deposit is really impossible, its too small. Do not let your contact be fooled into believing that if we place a small probe onto a solid sample we only obtain information from the point the probe strikes - not true!
3. If you do manage to stain the material with TEM type stains all you will get is a higher signal due to the metal being more emissive than an organic deposit, will this prove anything as the analytical problems remain the same?
Lets see what the MSA wizards come up with?
Good luck
Steve Chapman Senior Consultant E.M. Protrain, Oxford, UK Tel & Fax 44 (0)1844 353161
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
It is a little hard to imagine a scenario where the new crystal itself could be causing the copper counts (either directly or indirectly). Mind you, in the field of EDS systems, there are many wierd and wonderful things that can happen!
You say that you have put the detector back in original position. Do you mean in the axial direction? Is it possible, for example, that the snout has been bent slightly, so it is now at a different height? If so, then the collimator may be looking at a different view of the sample holder. It is also possible that the collimator has been installed incorrectly (or was perhaps incorrectly installed before and is now correct!) Perhaps even a different, less effective collimator was installed during the repair.
The hole count arises because of unfocussed radiation (both electrons and x-rays) which reaches the sample area, within view of the detector collimator. Anything which is hit by this radiation will generate characteristic x-rays. This would occur, for example, if your samples are mounted on copper grids or support rings. If parts of your sample holder are made of bronze (which is very common) then you may see copper originating from here. Another possible source might be an objective aperture blade which, though retracted, can still be hit by stray radiation. Do you still get the copper x-rays with the sample holder removed from the microscope? If so, then something else is being irradiated in the 'scope. Usually the objective polepieces are coated with carbon dag, or perhaps have a beryllium liner, so you dont see any characteristic x-rays from them, but even if they are bare, you would expect to see not copper but iron, perhaps with come cobalt (depending upon the alloy used in the polepieces).
Are you using a low-background sample holder for your x-ray work? This would have a beryllium insert to hold the sample, keeping the bronze parts further from the sample itself.
The purpose of the top hat aperture is to trap as many as possible of the x-rays and scattered electrons that arise from the edge of the beam-defining part of the aperture.
Different microscopes have widely different hole count characteristics, and as you didn't mention which model you are using it is not possible to give a hard number for what the hole count should be. In our instruments (which are VG's, known for their extremely low hole counts) the hole count would be around 0.1% of the count obtained from a reasonably thin area of the sample.
Good Luck,
Tony Garratt-Reed
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
Dear Robin, } } We've just had our EDS detector crystal replaced and I am now getting } complaints that the copper peak that is present whenever we collect } spectra (including hole counts) is too large. We've placed the repaired } detector at the same position as before and are using a Pt top-hat } aperture. My questions: } 1) What is the source of the Cu peak when we collect a hole count?
My guess is that either electrons or brehmsstrahlung x-rays from above the specimen are illuminating the (presumably copper) grid. When you replaced the detector did you also change the aperture? If not, then there should be no changes in illumination from before, so I can't account for any change in Cu peak.
} 2) How can I minimize the presence of the Cu peak?
Shielding both above the specimen to reduce stray electrons and x-rays and below the specimen to eliminate electrons backscattered from the objective pole piece will help. See my short communication in J. Elect. Microsc. Tech. 13:274-276 (1989) for how we did it.
} 3) How big of a Cu peak, is too big of a Cu peak? } If Cu is an element of interest, or if the Cu peaks overlap an element of interest, e.g. Os, Zn, any size is too big. If not, then if the Cu peaks cause significant dead time in the detector or significant continuum background, they're too big. You can measure the latter with "hole" counts where the "hole" is 1) a true hole and 2) a part of the specimen which contains the matrix (plastic, ice, glucose, etc. for bio- logical specimens, whatever surrounds the area of interest for materials specimens) but not the element of interest. 1) gives a measure of the stray radiation in the column, and 2) gives a measure of the stray radi- ation produced by the specimen. You can minimize 1), but you have to change the nature of the specimen to minimize 2). Good luck. Yours, Bill Tivol
We just received our new microtome unit. (Leica RM 2165) We wanted to prepare thin slice for optical and SEM observation. Suggestion on embedding media and staining procedure will be very appreciate. We have tried the disposable "WC" blade from Leica on section of 1" and we were able to get a good slice but I never relly look at this type of samples before. What is the best way to look at it: glue for glass slide and objectives for optical microscope. Thank's in advance. Line Mongeon ( Line is the french ways to right Lynn) Technologist premier. e-mail address: Mongeon-at-NTC.Noranda.com
Brigit, Try a smaller petri for spreading or two drops, interfernce colors will help with judging thickness. Personally I make my own from the parlodion sticks. Good luck.
Mike D JHMI Microscopy Facility
On Wed, 4 Feb 1998, Birgit Neubohn wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear microscopist, } } some weeks ago I asked for adresses for purchasing collodium. Thanks to all } who answered me! } Now I got pyroxilin (= collodium) 2% in amylacetate. } When I put one drop onto water, it spreads incredibly. So the resulting } film is much to thin and tears immediately when I get the grid into the EM. } Is the 2% solution to weak? } Is there another method to produce a thicker film? } } I am trying the collodium support for I get enormous binding of the } *primary* antibodies to the formvar sometimes. I find it often, that } primary antibodies bind to formvar, but not to that extend that I saw with } two antibodies from eggyolk and rabbit. Any suggestions how to overcome } this? } } Thanks a lot in advance } } Birgit } } } Dr. Birgit Neubohn } Institute of Plant Genetics and Crop Plant Research (IPK) } Corrensstr. 3 } D-06466 Gatersleben-Deutschland } } Tel.: (+49) 039482 5447 } Fax: (+49) 039482 5139 } e-mail: neubohn-at-ipk-gatersleben.de } } } }
If you are considering ZIP discs / drives (which are great) also consider JAZZ discs / drives, which are essentially the same, but hold much more data, and images take up data storage space. Also you may wish to look into a CD recorder unit, which is also a great way to store digital images. Although no digital image can compare to the resolution of film, not yet anyway. best of luck -Mike
On Tue, 3 Feb 1998, Paula Sicurello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Listers, } } I know a lot of you are real smart when it comes to computer images } & stuff. I have an optical disc drive on my SEM and would like to go to } something that most people have (we have the only optical disc drive on } campus). Folks here have suggested Zip Drives. Now, I'm an idiot when it } comes to computer stuff, is a Zip drive good enough for storing images and } will people get decent images back when they put them on their lab } computers? } I've heard that Zip drives and the discs are fairly inexpensive, is } that true? } } Thanks in advance for all your fine help. } } } Still preferring to make photographs, } } } Paula = ) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } } }
Richard- one way of isolating equipment (in AFM and other scanning probe microscopies) which may or may not work for your situation is to suspend the equipment from the ceiling, designing the damping system into the suspension system...? -Mike
On Tue, 3 Feb 1998, Richard Fonda wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear list members, } } I am looking for suggestions on features we should consider for an EM lab } renovation. We are currently in the design process for renovating a } building which will house two TEMS, 2 SEMs, an AES, and a darkroom. With } this opportunity to design the EM labs, we want to take all reasonable } precautions and make the necessary improvements to optimize these areas. } This includes necessities for EM operation as well as conveniences. } } While this currently unoccupied building readily passes vibrational and } magnetic field tests, I am trying to minimize the impact of the labs, } offices, and electrical/ventilation systems which will surround the EM } labs. Alderson's book (suggested on the list a while back) was a great } help for the initial design stages. What suggestions do you have either } for the design of the laboratories or for the related equipment? While my } primary interest is for the TEM labs, I would welcome any suggestions for } the other EM rooms or dark room as well. } } One major concern I have is with mechanical vibration isolation. We would } like to limit a priori the effects of the surrounding labs and services. } Any suggestions which would limit the effects of mechanical vibrations at } the TEMs, or reduce the level of ambient vibrations, would be particularly } helpful. } } Richard Fonda } } _____________________________________________________________ } Richard W. Fonda Naval Research Laboratory } (202) 767-2622 Code 6324 } (202) 767-2623 fax Washington DC 20375 } _____________________________________________________________ } } }
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Regarding Image Analysis Course
There are actually two image analysis courses offered at NC State, one by = John Russ and one by John Mackenzie. I have taken both the courses and both are good but for different = purposes.
John Russ' courses deals mostly with image measurement and what one needs = to do to make that possible. John MacKenzie's course is an excellent basic course in what digital = imaging is all about, how to make and print good looking photographs, = scanning images, how one needs to set up to do digital imaging, etc.
For a basic digital imaging course, John Mackenzie's course is more tuned = in to making good looking images and the digital basics. John Russ' is really more advanced dealing with doing image measurement, = manually and by computer.
Just my thoughts for what they are worth. Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
Regarding John Russ' reply: I am a graduate of the NCSU course and readily recommend it to anyone who is looking for a solid foundation in digital imaging. I hope my response is not as "suspect" as John's -the only "kickback" I get from the course is the knowledge!
Cheers
Bill Heeschen The Dow Chemical Company
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Birgit, we routinely use 2% collodion films cast on distilled water. You didn't mention carbon coating your films after they were placed on grids. This is necessary to dissipate the energy from the beam. I try not to use films if it can be avoided. However, with troublesome plant or bacterial samples it is often needed. Hank Adams Electron Microscopy Lab New Mexico State University Las Cruces,NM 88003 phone: 505-6463600 fax: 505-6465665
I know this may be "species specific" but we use a magneto-optical (MO) drive system with our JEOL 5800LV SEM for image storage. The disks are 260MB-acres of space for bitmap or tif image storage. I don't know if MO drives are not as readily available as rewritable CD's but thought I'd mention them.
Tracey Pepper Bessey Microscopy Facility Iowa state University
Look in the tutorial section of the disk and you will find a good instruction and tutorial on the plug-ins. There are also the same images that they use in the manual so that you can try things out yourself. They are Adobe ".pdf" files and there is a reader on the CD. You should also consider getting John Russ' companion book, "The Image Processing Handbook". The URL for the website is http://members.aol.com/ImagProcTK/index.htm
John has info on a short course at http://vims.ncsu.edu/matsci/IPCFacul.html
and his email address is john_russ-at-ncsu.edu
He has answered questions for me in the past and he has addressed questions on the listserver as well.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
we recently bought an Image Processing toolkit with plug-ins for Photoshop. It runs under Mac as well as Windows and has, among a lot of other things, FFT. The package is from Reindeer Games. I haven't tested it very severely but it did give me good power spectra. Unfortunately the documentation is rather limited and I can't find their full address.
Hope this helps anyway.
Nick Schryvers
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We are using an Agfa Duoscan (1000 dpi) for digitizing HRTEM images.
At the moment we are saving the images to disk, transfering the tif files via Apple File Exchange to the Mac, then using Digital Micrograph to give us an FFT, for printing/exporting etc.
Question:
Is there software available, (freeware or...) that would allow us to perform the "FFT" on the "PC" that we are using to digitize the HRTEM?
Thanks in advance
Fred Pearson Electron Optics Coordinator
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
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There will be a demo of the Hitachi variable pressure SEM in conjunction with the joint FL AVS and Florida Society for Microscopy meeting at the University of Central Florida in Orlando February 23-25, 1998.
For more information or to set up a demo appointment please contact me directly. You my bring you own samples to the demo.
************************************************************************* Lucille A. Giannuzzi, Ph.D.
Assistant Professor Dept. of Mechanical, Materials, and Aerospace Eng.
Director, Cirent/UCF Materials Characterization Facility President, Florida Society for Microscopy
University of Central Florida phone (407) 823-5770 PO Box 162450 fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA ----------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain *************************************************************************
most e.m. units are at ground floor or basement level so consider the risk of flooding. This may be natural flooding (when the drains block with leaves in autumn/fall), the lab above may often flood or someone leaves the tap on in the darkroom. Precautions may range from a pair of wellies and sandbags to sills and sealable doors (I've seen something like that in a London University). But at the very least it should be possible to isolate the electrics to microscope and lab in an emergency.
Flooding is less likely than vibration and magnetic fields but it does happen and can be disastrous.
Malcolm Haswell e.m. unit University of Sunderland UK ----------
Dear list members,
I am looking for suggestions on features we should consider for an EM lab renovation. We are currently in the design process for renovating a building which will house two TEMS, 2 SEMs, an AES, and a darkroom. With this opportunity to design the EM labs, we want to take all reasonable precautions and make the necessary improvements to optimize these areas. This includes necessities for EM operation as well as conveniences.
While this currently unoccupied building readily passes vibrational and magnetic field tests, I am trying to minimize the impact of the labs, offices, and electrical/ventilation systems which will surround the EM labs. Alderson's book (suggested on the list a while back) was a great help for the initial design stages. What suggestions do you have either for the design of the laboratories or for the related equipment? While my primary interest is for the TEM labs, I would welcome any suggestions for the other EM rooms or dark room as well.
One major concern I have is with mechanical vibration isolation. We would like to limit a priori the effects of the surrounding labs and services. Any suggestions which would limit the effects of mechanical vibrations at the TEMs, or reduce the level of ambient vibrations, would be particularly helpful.
Richard Fonda
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375
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The idea to copy a "batch" of zip disks to CD-R is very good. Do beware that some sources cannot feed data fast enough to a CD-R (especially =} 2x) and will cause a "buffer under run" at the CD-R. This will make a frisbe rather than a data disk. A SCSI Zip may be fast enough, but it should not be assumed that it will work (I have not tied). A solution is to assemble the intended DC-R contents on a HDD 650 Meg partition. A fast SCSI HDD is best, but I have had only one buffer under run failure using my P166 with a EIDE/WD3.2 HDD.
Woody White McDermott Technology, Inc. http://www.mtiresearch.com
Home: woody.white-at-worldnet.att.net http://www.geocities.com/capecanaveral/3722 (Sorry about the $%#* cookies and browser openings that randomally hit - but it is free!)
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One idea is to get both a Zip Drive and a CD read/writer. Store your images on the Zip disks and when you get six to eight filled up, transfer the files to a CD. Then recycle the Zips. The Zip drive is more agile when working with individual files. Writing to the CD is best done in a few (6 to 8 if you are copying from 6 to 8 Zip disks) sessions. Using the Zip disks allows you to make changes before archiving the final file(s). Often a lab or department with several computers used in image handling will have Zip Drives on each computer, and a single CD writer that is external & portable. This arrangement will give you the write/read/change/write again flexibility of the Zips and the efficient & permanence of the CDs. And you won't have to buy truck loads of Zip disks......just a car load of CDs.
Joiner Cartwright, Jr., Ph.D. Assistant Professor of Pathology Baylor College of Medicine Houston, Texas U.S.A.
id AA40230; Thu, 5 Feb 1998 09:07:58 -0500 Received: from CASSERVER1/SpoolDir by casmail.muohio.edu (Mercury 1.32); 5 Feb 98 09:07:58 -5 Received: from SpoolDir by CASSERVER1 (Mercury 1.32); 5 Feb 98 09:07:31 -5 To: microscopy-at-Sparc5.Microscopy.Com
I should have mentioned the following in my post yesterday:
When casting Collodine on water it is important that you use a large enough container so that the solution can spread to its full extent and not be limited by 'running into' the sides of the container. When it hits the side(s) of the contianer the film will pileup in that area resulting in an uneven film thickness.
Any circular dish seems to work, but I prefer using 10 -12" pyrex baking dishes purchased from local home stores (i.e. K-mart, Wallmart, etc.). They are very cost effective ($3-9 US), and have nice thick walls which stand up nicely to general lab abuse.
Good casting again!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"CONGRESS.SYS Corrupted: Re-boot Washington D.C. (Y/N)?"
BUEHLER also offers an Image Analysis course two times a year. The first of these has already concluded, but space is available for our summer course. BUEHLER is a manufacturer of Image Analysis Systems, but, although we do use BUEHLER products to conduct the class, this course is not focused on sales. It is a technical description of the methods and mechanisms of Image Analysis for Materials Science.
The following is a description of our class as printed in our course schedule:
PRINCIPLES AND PRACTICE OF IMAGE ANALYSIS Instructor: M. Hoffmann Quantitative analysis of specimens is invaluable for the development and maintenance of a quality product. Principles of Image Analysis is an introductory course teaching the basic theory and practice of image analysis. The course provides hands-on experience and will cover many common measurements such as nodularity, area percentage of constituents, coating thickness, and ASTM E112 grain size. No experience in image analysis is required, but an understanding of microstructural evaluation, specimen preparation, and computer literacy is assumed. 2.2 CEUs Jan. 20-22 Irvine, CA Aug. 17-19 Lake Bluff, IL
A wide selection of other materials related courses are available. Please see this list on our Web Site (http://www.buehlerltd.com) or call for a course schedule.
For more details on course availability and prices, please contact Sandy Kaucic at (800)323-9330, Ext. 4679.
Regards, Scott D. Holt BUEHLER, LTD PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 USA (847)295-6500, Ext. 4546 http://www.buehlerltd.com
The abstract deadline has been extended to Friday February 13th for the Microscopy and Microanalysis '98 meeting to be held in Atlanta GA from July 12-16th, 1998. Please do not delay. Get your abstracts in as soon as possible. Check out the website at :
for more details on planned symposia as well as abstract submission.
Kathi
Kathleen B. Alexander Metals and Ceramics Division Oak Ridge National Laboratory P.O. Box 2008 MS-6376 Oak Ridge, TN 37831-6376 PH (423) 574-0631 FAX (423) 574-0641
I have a student that is trying to deterimine the grain size of a metal that has twins. He is using the line intercept method and is wondering how he should treat the twin bondaries. It isn't always clear what is a twin and what may be a grain. Does anyone know how to correct for this or how twins are treated in this type of analysis. Thanks.
Roberto Garcia EMF Manager Wright State University
If you become interested in CDs: Rewritable (CD-RW) is the latest form of CD storage. Look for the newer CD-RW drives with "packet writing" capability. You will still need a compatable disk drive and software. For extensive information check out the web sights for some of the 'brand' name CD-RW devices. You can find these names in recent computer products sales flyers etc.
Original message was:
} I know a lot of you are real smart when it comes to computer images } & stuff. I have an optical disc drive on my SEM and would like to go to } something that most people have (we have the only optical disc drive on } campus). Folks here have suggested Zip Drives. Now, I'm an idiot when it } comes to computer stuff, is a Zip drive good enough for storing images and } will people get decent images back when they put them on their lab } computers? } I've heard that Zip drives and the discs are fairly inexpensive, is } that true?
} Thanks in advance for all your fine help.
} } Still preferring to make photographs,
} Paula = )
} Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
Just to remind those of you still in shock after the SuperBowl, the deadline for applications to the UBC "Live Cell" course is at the end of this month. Enrollment is limited but there is still room.
Basic information about the course is to be found below. Many more details including the complete course brochure, this year's tentative Program Outline, some 3D results from last year's course and AN APPLICATION FORM (!) can be found at:
10-Day Short Course on 3D Microscopy of Living Cells
June 17 - 28, 1998
and
Second, Post-course Workshop on
3D Image Processing June 30 - July 2
in association with the UBC BioSciences Microscopy Facility and the Department of Computer Science
University of British Columbia Vancouver, BC, Canada
Organized by Prof. James Pawley University of Wisconsin-Madison
=46aculty
* Jon Art University of Illinois * Pin Ching Cheng State U. of New York, Buffalo * Rachel Errington University of Nijmegen * Elaine Humphrey University of British Columbia * Jim Pawley University of Wisconsin-Madison * Ernst Stelzer EMBL, Heidelberg * Michael Weis Agriculture Canada * Nick White Oxford University * Dan Focht Bioptechs, PA * Ted Inou=E9 Universal Imaging, PA * Larry Keenan Cell Robotics, NM * Paul Millard Molecular Probes, OR * Sigrid Myrdal Multidimensional Imaging, WA * Paul Negulescu Aurora Biosciences, CA * Hans Van der Voort Scientific Volume Imaging, NL
TUITION
Course tuition is $1,950 US and includes lunches. On receipt of 50% deposit, all students will receive preliminary group assignments and a copy of the textbook, Handbook of Biological Confocal Microscopy, (Plenum, 1995). The tuition fee includes single tickets for the Opening Reception, the Manufacturer's Reception and the Beach Party, the textbook and all handouts. Accommodations and meals other than lunch are not included in the tuition fee.
APPLICATIONS
Applicants will complete a questionnaire to assess knowledge level and field of interest. Enrollment will be limited to about 24 participants. Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with basic texts on request to read before the course begins. Application forms can be down-loaded from the WWW site at
Prof. James Pawley, Rm. 1235, 1500 Johnson Dr., Madison, WI, USA 53706. Phone: 1-608-263-3147, Fax 1-608-265-5315, Email: jbpawley-at-facstaff.wisc.edu
Application deadlines:
Application forms must be received by March 1, 1998!
Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 1998 to reserve your position. In general, refunds of the deposit will only be possible if your position can be filled from the Waiting List. The remainder of the fees are due before registration.
DATES:
Applications must be received by Mar. 1/98 deposit due Apr. 15/98 Registration 8:00 - 7:00 pm Wednesday, June 17/98 Last class will end with lunch Sun., June 28/98
*******************************
3D Image Processing Workshop
June 30 - July 2, 1998
The course is designed for biologists who need to make measurements on 3D microscopical data sets and then display the results in an effective manner. The course will be taught in a computer laboratory belonging to the Computer Sciences Department at the University of British Columbia which contains 27 SGI Indy workstaions and much of the other equipment needed for the measurement and display of 3D digital image data. Software from a variety of vendors serving the 3D microscopy market will be described, demonstrated and available for use.
Course Organizers
* Nick White Oxford University * Hans Van der Voort Scientific Volume Imaging, NL
=46aculty * Pin Ching Cheng State U. of New York, Buffalo * Rachel Errington University of Nijmegen * Alain Fournier Computer Science, UBC * Sigrid Myrdal Seattle, WA
Tuition (including lunch) $700 (US)
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Engineering Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr.,
I have a CCD attatched to the bottom of my Philips EM430 that, over the years, has acquired a bit of dust, lost samples, Be rings, etc... I want to be able to clean the surface off before I start my in-situ work, but am a little tentative about doing this. Is there anything I should / shouldn't do if I want to keep my job, i.e. what are the best ways to destroy a CCD from the inside? What would be the best method of going about this? Should I remove the whole camera or is it best to do this inside the column? Thanks in advance for any help.
Brian Gorman bgorman-at-umr.edu Graduate Research Assistant (573) 341-4405 Electronic Materials Applied Research Center 303 Materials Research Center University of Missouri - Rolla Rolla, MO 65409 http://www.umr.edu/~bgorman
According to my ancient copy of DeHoff and Rhines, "Quantitative Microscopy," grain size is determined per phase. As twins are crystallographic phenomena within the grains of any given phase they should not be counted. You can use a linear intercept method to measure mean twin intercept distance if you like, but don't confuse the twins with the grain boundaries.
This explanation makes intuitive sense as well. Dislocations are a crystallographic phenom within the grains of any given phase but you wouldn't count dislocations along with grain (or twin) boundaries when measuring *grain* size.
The different diffraction contrast shades of black and white, as a function of tilt, for individual grains, the b/w contrast of twins, and the odd dislocation seen in a TEM of a polycrystalline material are the main reason that video based grain size measuring machines *don't* work! (It's been a while since anyone kicked that sleeping dog)
Hello all, An associate is interested in purchasing a basic second hand SEM with EDS for his private consulting (glass technology) company. If you know of available instruments, I welcome you to contact me privately. Dealers welcome. Thank you in advance for the information.
Regards, Mark S. Angelone Penn State University Materials Characterization Lab 814-865-0344 angelone-at-geosc.psu.edu
I need a protocol for the immunogold labelling of Golgi membranes deposited on grids covered with 1% formvar and stained with Uranyl acetate. So far I have tried several conditions by using as primary antibody the monoclonal one directed against the cytoplasmic tail of Giantin: but unsuccesfully !!
Dr. Roberto Weigert Consorzio Mario Negri Sud Department of Cell Biology and Oncology Molecular Neurobiology Laboratory Via Nazionale S.M. Imbaro, 66030 Chieti Italy Phone: 0039-872-570-354 Fax: 0039-872-578-240
If you consider a JAZZ drive it is cheaper to buy a newly introduced Syquest SparQ which has a capacity of Jazz drive at about the price of a Zip drive.
Ann Fook
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Agriculture Agri-Food Canada, Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
} } } MIKE ROCK {merock-at-du.edu} 02/04/98 04:55pm } } }
If you are considering ZIP discs / drives (which are great) also consider JAZZ discs / drives, which are essentially the same, but hold much more data, and images take up data storage space. Also you may wish to look into a CD recorder unit, which is also a great way to store digital images. Although no digital image can compare to the resolution of film, not yet anyway. best of luck -Mike
On Tue, 3 Feb 1998, Paula Sicurello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Listers, } } I know a lot of you are real smart when it comes to computer images } & stuff. I have an optical disc drive on my SEM and would like to go to } something that most people have (we have the only optical disc drive on } campus). Folks here have suggested Zip Drives. Now, I'm an idiot when it } comes to computer stuff, is a Zip drive good enough for storing images and } will people get decent images back when they put them on their lab } computers? } I've heard that Zip drives and the discs are fairly inexpensive, is } that true? } } Thanks in advance for all your fine help. } } } Still preferring to make photographs, } } } Paula = ) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } } }
Grain size measurement is the subject of two ASTM standards. Standard E 112 describes manual methods, and even has an adjunct comparison chart showing twinned grains. Grain size by image analysis is the subject of ASTM standard E 1382. The latest version was written in 1997, and both standards can be found in ASTM Annual Book of Standards, Vol. 03.01. Some version of the Book of Standards can usually be found in the library. In E 1382, Section 6.3 discusses twin boundaries, and as Ron Anderson said, they should be ignored. A thorough discussion of this topic can be found in an article by George Vander Voort (1984), in which he states, "When measuring the grain size of austenitic metals, annealing twins are ignored." If one is using a completely automatic method, it is necessary to use software to remove the twins. E 1382 calls it "image amendment techniques." An article on one such an approach is in ASTM STP 1165.
G. F. Vander Voort, "Grain Size Measurement," Practical Applications of Quantitative Metallography, ASTM STP 839, McCall and Steel, eds., ASTM, Philadelphia, 1984, 85-131.
J. J. Friel and E. B. Prestridge, "Artificial Intelligence for Twin Identification," Metallography: Past, Present, and Future, ASTM STP 1165, G. F. Vander Voort, et al eds., ASTM, Philadelphia, 1993, 243-253.
At 10:03 AM 2/5/98 -0400, Kathi Alexander wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Kathi,
It was my understanding that the MSA dates had been changed to the week of July 26. Please clarify!
Barbara Foster Microscopy/Marketing & Education 125 Paridon Street, Suite B Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
REDUCED FEE REGISTRATION DEADLINE January 31, 1998 Workshop on Tripod Polishing
Workshop Objective This course will cover all aspects of pre-thinning and focus on final thinning for TEM via Tripod Polishing. Due to the limited class size an= d the extensive hands-on opportuinities, this course is well suited to novices as well as advanced Tripodders. Attendees will also learn the latest techniques available in ion milling and in plasma cleaning for TEM=
samples. The course will include sections on:
How to do it and why should I? What's really going on and what am I really seeing? How to prepare small, specific area cross-sections. The problem of wildly differing materials (eg tungsten). Rapid preparation of TEM cross-sections. Preparation of a wide range of materials: semiconductors, ceramics, metals,...
Hands-on Opportunity This course will be unique in that it will provide a hands-on opportunity=
for every class participant. Tripod Polishers, Polishing Wheels, and pre-thinning equipment will be made available to all participants and actual samples will be prepared - by the students - as part of the course= =2E =
This is a great opportunity to get your hands dirty and actually learn by=
doing. The instructors will walk you through each step of the process an= d then let you loose on the equipment. This course is designed to teach th= e Tripod Polishing technique. Silicon samples will be provided to the students and used as the basis for the course teaching.
Workshop Location and Dates South Bay Technology - San Clemente, CA Dates: Friday & Saturday - March 13 & 14
Previous Participants (partial list) INTEL, AMD, Motorola, LSI Logic, Conner Peripherals, Univ of Maryland, Un= iv of New Mexico, UNAM (Mexico), LG Electronics (Korea), Battelle, MEMC, MVA=
Inc., Univ of Michigan, U.S. Bureau of Mines, IBM, Naval Research Lab, Purdue Univ, Univ of Alabama, Univ of Arizona, Univ of Colorado, Univ of Wisconsin.
Class Size Due to the intensive hands-on aspects of this course, class size will be strictly limited to 10 participants.
Registration Fee: $795 (includes lunches and Friday night Dinner) $695 if registration fee paid by January 31, 1998=
Registration Deadline: 30 days prior to workshop
For additional Information: Monica Pflaster South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 TEL: 800-728-2233 FAX: 714-492-1499 e-mail: sbt-at-southbaytech.com
ON-LINE Registration available at: http://www.southbaytech.com
Registration Form
To register for the workshop, please fill out this form and send it, with=
registration fee to:
South Bay Technology, Inc. =
Workshop on Tripod Polishing 1120 Via Callejon San Clemente, CA 92673 USA
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} } I know a lot of you are real smart when it comes to computer images } } & stuff. I have an optical disc drive on my SEM and would like to go to } } something that most people have (we have the only optical disc drive on } } campus). Folks here have suggested Zip Drives. Now, I'm an idiot when it } } comes to computer stuff, is a Zip drive good enough for storing images and } } will people get decent images back when they put them on their lab } } computers? } } I've heard that Zip drives and the discs are fairly inexpensive, is } } that true? } } } Thanks in advance for all your fine help. } } } } } Still preferring to make photographs, } } } } Paula = ) } } } Paula Sicurello } } UC Berkeley } } Electron Microscope Lab } } psic-at-uclink4.berkeley.edu } Sorry I am a bit slow to respond to this one, but it is of particular interest to me.
I just got back from visiting a friend on the west coast that deals with commercial imaging, and I have some insightes you may all be interested in. If these things have been said already I apologize in advance.
In Regaurds to storage of images:
There are two (and many more) important things to consider, storage space and access.
CD-Rom Pro's
A CD-ROM offers alot of storage space (up to 640 MB). Most PC's and Mac's these days have a CD rom reading device (or one can be cheaply added). Cd-ROMs are not mag-media and thus are safer in magnetic fields. The life time of a CD ROM is ~50 years in atmosphere and estimated over 100 in an inert atmosphere (vacuums are actually bad for CD-R's, they can cause "crazing" (sp?) of the read surface). Blank CD's are down to about $2 now.
CD-R con's
A single write CD-R presents the problem that, unless one has a large number of images, alot of the CD is wasted in writing. Further, Single write CD's do not alow a user to modify a file after it is written to the CD. Packet writing can help. It allows the user to sequencially write packets to the CD. The problems faced here are:
The computer used to read the packeted cd must have software that allows it to do so....UNFORTUNATLY, companies that put out CD-R writers have not come up with a standard - This is a major "CON"
Multiple write CD-R's are better. They allow a limited number of rewrites to the CD. This allows for some modification of a CD. There are still compatability issues with these CD devices as well as a question about CD-R life time.
One major CON to cd-r's is the cost of a writing drive. They start at ~$300 and go as high as ~$1000.
ZIP DRive Pro's
Welcome to the floppy of the future!! A ZIP disks holds about 100 MB and has a life time of 5-10 years (estimated). I currently purchase them for $12 each (I have seen them as high as $20). Zips are fully rewritable (they are basically just like floppies). Many new computers come with ZIP drives, new ones can be added for $100-300 (depending on the configuration of your computer system). My friend's imaging company no longer even uses the old 3.5" floppies..."They are usless," in his words. ZIP drives are portable. At the Institute I work at we have 3 ZIP drives that can be checked out and put on nearly any of our computers. We don't need a zip for each one, just a port (though I would very much like my own). ZIP disks have far fewer compatibility issues than CD-Rs. To deal with compatability here, all of our ZIP's are PC formatted. This is because our Macs can all read PC disk (as well as our Pc's) but not vice versa.
ZIP Drive CON's
Zip dizks are magnetic media and are thus damaged by X-rays and EM fields. They have shorter lifspans than CD-R's. ZIP disks hold 1/6 what a CD-R holds. They cost more than a CD-R.
Advice from an Imaging Company:
My Friend's Company (and I am triing to move in this direction myself) does the following, ZIP disks are used for file transfer (the images they deal with are huge and keeping them on disk is easier than tranferring them over the network). ZIPs are NOT used for archiving, only as working copies. Then when a project is finished, it is burned onto a CD-R. To avoid compatibility problems they have devised an archiving system.
There's my 2cents...for what they are worth.
Image analysis and transfer on ZIP Long term storage on CD-R
Christopher (P.S. Forgive the grammer and spelling errors....I'm in a hurry)
I wholly agree with Ron Anderson. I have only seen reliable automatic identification of twins in cases where the grain size is large relative to the distance between grain boundaries. Using manual counting techniques allows one to evaluate whether a particular boundary is a grain or twin boundary. Unfortunately, that distinction can also be difficult. A couple distinguishing features of twins:
1) twins tend to be linear or have linear segments corresponding to the coherent twin boundary orientation 2) the low interfacial energy of twins does not cause a large deflection in the intersected grain boundary, which is typically caused when a grain boundary (usually high energy) intersects another grain boundary.
If the equipment is available, I believe that grain sizes can also be determined while excluding twins with EBSP in an SEM to map the grain orientations across a region.
Good luck, Richard Fonda
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Wouldn't it be easier for everyone to read if messages announcing workshops, conferences, meetings, etc., contained a reference to place (country, state) in a subject line?
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by caesar.wits.ac.za (8.8.7/8.8.5) with ESMTP id IAA05339 for {MICROSCOPY-at-MSA.MICROSCOPY.COM} ; Fri, 6 Feb 1998 08:00:29 +0200 (GMT) Received: from GECKO/SpoolDir by gecko.biol.wits.ac.za (Mercury 1.21); 6 Feb 98 07:59:01 GM+2 Received: from SpoolDir by GECKO (Mercury 1.21); 6 Feb 98 07:58:33 GM+2
I am analysing bulk alloys of high atomic number elements eg. Re, Ir (Z=75, 77) in a light element matrix eg Al (Z=13) at 20 kV with an EDS system in an SEM using ZAF corrections and pure element standards. Typically, the high Z elements have L alpha peaks at around 9kV, M alpha peak at around 2kV. The high Z content ranges from 0.1 to 50 at%.
As I understand it, correction factors for L lines are better than for M lines. However, the M peak is much higher than the L peak at 20kV, so statistics are better. At the low concentrations of high Z element, I only see an M peak, not L peak. There is about up to 2 at% difference analysing using both peaks over the alloy composition ranges and such a difference is a problem for our work. Unfortunately I do not have any alloy standard to check results against! So which peak to use? Yes, someone analysed on a microprobe (using WDS) utilising same area on a sample and same standards and got a third answer which gave no direction to my question!
Any advise would be gratefully received. Thanks in advance. Mike
Michael J Witcomb PhD Electron Microscope Unit University of the Witwatersrand Private Bag 3 WITS 2050 South Africa
I suppose that would make it easier for people to delete messages in whic= h they have no interest. I apologize for any inconvenience. I never real= ly thought about there being geographical boundaries for the workshop as we have had people attend the workshop from throughout the USA, Asia, Europe= , Africa etc. =
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Wouldn't it be easier for everyone to read if messages announcing workshops, conferences, meetings, etc., contained a reference to place (country, state) in a subject line?
} } I have a CCD attatched to the bottom of my Philips EM430 that, over the } years, has acquired a bit of dust, lost samples, Be rings, etc... I want } to be able to clean the surface off before I start my in-situ work, but am } a little tentative about doing this. Is there anything I should / } shouldn't do if I want to keep my job, i.e. what are the best ways to } destroy a CCD from the inside? What would be the best method of going } about this? Should I remove the whole camera or is it best to do this } inside the column? Thanks in advance for any help. }
I've heard You shouldn't use a "Dust Off" or similar because such a strong stream of air could damage the scintillator. I've seen an expert use a little pump that looks and functions very much like a syringe, but I guess a clean plastic syringe with a biggish hole should do (without needle). In any case use it gently. (Never touch the scintillator with anything hard or with Your fingers).
You might also have oil dropletts on the scintillator. You can use certain solvents and materials to wipe the surface, but You should definitely ask the manufacturer about that.
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
From recent experience, I advise that anyone buying a new CD-RW system consider the following:
You should be aware that CD-RWs have low reflectance compared to CD-R media. The consequence of this is that CD-RWs CANNOT be read using many 'older' CD-ROM drives which will happily read CD-Rs. If you are using the media for archiving, then CD-Rs are safer anyway (also cheaper).
"Packet writing" is another good way or writing discs that others may not be able to read! I know this may sound a bit quaint to some, but we are still using Windows 3.11 and any disc written with packets is unusable in this environment. It's fine for a Windows95 or NT world but if your data is to be really portable, avoid packets.
---------------------------------------------------------------------- Simon Dumbill, AEA Technology plc, 220, Harwell Didcot Oxfordshire OX11 0AB UK
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If you become interested in CDs: Rewritable (CD-RW) is the latest form of CD storage. Look for the newer CD-RW drives with "packet writing" capability. You will still need a compatable disk drive and software. For extensive information check out the web sights for some of the 'brand' name CD-RW devices. You can find these names in recent computer products sales flyers etc.
Original message was:
} I know a lot of you are real smart when it comes to computer images } & stuff. I have an optical disc drive on my SEM and would like to go to } something that most people have (we have the only optical disc drive on } campus). Folks here have suggested Zip Drives. Now, I'm an idiot when it } comes to computer stuff, is a Zip drive good enough for storing images and } will people get decent images back when they put them on their lab } computers? } I've heard that Zip drives and the discs are fairly inexpensive, is } that true?
} Thanks in advance for all your fine help.
} } Still preferring to make photographs,
} Paula = )
} Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
Fri, 6 Feb 1998 14:09:49 +0000 Received: from localhost (rdoole-at-localhost) by ermine.ox.ac.uk (1.1/8.8.3) with SMTP id OAA29239 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 6 Feb 1998 14:09:42 GMT
Hi,
Are there any users out there with experience using the Gatan GIF with a 2Kx2K MSC camera? If so please contact me (off list) as we are interested in finding out practical information about its use, data handling etc.
Thanks, Ron
========================================================================== = Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ========================================================================== ==
I seem to recall a company named "Discryptic Designs" located in Seattle, WA that was marketing two products called "Pattern Isolator" which is a simple pc based FFT plug-in for Adobe Photoshop. The company also had another plug-in product called "Color Isolator".
I no longer have an address for Discriptic Designs but perhaps the phone company can provide a number where you can contact them to obtain further information.
Dan Purdy Ottawa, Canada tel: (613) 741-8939 fax: (613) 741-0511
At 05:00 PM 2/3/98 -0500, you wrote: } ------------------------------------------------------------------------} Qu estion: } } Is there software available, (freeware or...) that would allow us to } perform the "FFT" on the "PC" that we are using to digitize the HRTEM? } } Thanks in advance } } Fred Pearson } Electron Optics Coordinator } } ******************************************************** } Fred Pearson } Brockhouse Institute for Materials Research } McMaster University } 1280 Main St. West } Hamilton, Ontario } Canada L8S 4M1 } } ********************************************************
Zip Drives/Discs win in the temporary storage category.
CD-ROM(the writeable CD's) win in the archiving category. Someone said make sure and use the gold CD's.
Honorable mention: SyQuest SparQ (capacity of a Jazz, price of a Zip)
Dis-honorable mention: Jazz Drives (apparently have a lot of problems)
Sad, but true addendum- After all this great advice from you guys, it turns out my imaging computer is too old & slow to even work a Zip drive. I was going to Zip for temporary storage, and optical for archiving. And, as always, there's no money to upgrade. SIGH!
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Dear colleagues! I need analytical formula (or formulas) for dependense intensity of unelastic scattered electrons from propagation vector (wave angle). It must take into account temperature factor (Debye-Waller), plasmon scattering etc. My case is 300-600 A (angstrom) amorphous films. Thank you very much for attention to my question. With best wishes to all Alexsandr Domantovski RRC "Kurchatov Institute" Moscow, Russia.
by dogwood.botany.uga.edu (8.8.8/8.8.8) with SMTP id OAA16707 for {Microscopy-at-Sparc5.Microscopy.com} ; Fri, 6 Feb 1998 14:51:02 -0500 (EST) Message-Id: {199802061951.OAA16707-at-dogwood.botany.uga.edu} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Brian,
We had a CCD camera set up like you described and when it needed cleaning the Gatan man did the job. He removed the camera and gently flowed acetone (or some solvent) over the scintillator to rinse the oil, dust, whatever, off. It worked. I wasn't brave enough to do the cleaning (a CCD is a mega expensive item if ya mess up). I recommend that you contact the manufacturer and get their advice. good luck, beth
} Brian Gorman wrote: } } } } } I have a CCD attatched to the bottom of my Philips EM430 that, over the } } years, has acquired a bit of dust, lost samples, Be rings, etc... I want } } to be able to clean the surface off before I start my in-situ work, but am } } a little tentative about doing this. Is there anything I should / } } shouldn't do if I want to keep my job, i.e. what are the best ways to } } destroy a CCD from the inside? What would be the best method of going } } about this? Should I remove the whole camera or is it best to do this } } inside the column? Thanks in advance for any help.
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Mike Rock's suggestion to suspend an instrument from the cieling using springs is indeed a feasible approach. I visited a laboratory in China this summer where the vibration problems in an old building were handled in exactly that way. Their instruments were all sitting on platforms which were suspended from the ceiling by groups of eight or ten springs (each spring about one inch in diameter and 5 feet long) attached at the corners, and at other strategic locations around the platforms as needed to compnesate for the weight distribution, and anchored in the ceiling. Small adjustments needed to keep the platforms level were achieved by varying the number of springs in the groups, and by varying the tension on some of the individual springs by means of turnbuckles. I may have a picture of the setup, if anyone is seriously interested.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
It would seem to me that the best way to avoid the possibility of having an instrument lab flooded would be to have an ample drain in the floor of each lab room. If you don't trust the local sewer or drain system, then a sump pump could also be installed. A word of caution, be sure that the cement-workers who do the floors are sober during the process; otherwise, you're likely to end up with the situation I have in the basement of my house - there's a lovely drain, but the floor around it has the contour of a volcano, and so water can't spontaneously reach the drain.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
I need analytical formula (or formulas) for dependense intensity of unelastic scattered electrons from propagation vector (wave angle). It must take into account temperature factor (Debye-Waller), plasmon scattering etc. My case is 300-600 A (angstrom) amorphous films. Thank you very much for attention to my question. With best wishes to all Alexsandr Domantovski RRC "Kurchatov Institute" Moscow, Russia. ------------------------------------------------------------------------ Lenz (Z Naturforsch. 91, 1954, 185-204) provided an atomic formula for the inelastic angular distribution which seems to work quite well for amorphous films. It does not include phonon scattering. An angle-integrated version of this formula is given in Egerton, "EELS in the Electron Microscope", 2nd edition (Plenum, 1996), and also a computer program which includes plural scattering.
Ray Egerton, Physics Dept, University of Alberta, Edmonton, Canada T6G 2J1 Phone: 403-492-5095, FAX: 403-492-0714, e-mail: egerton-at-phys.ualberta.ca ------------------------------------------------------------------------
I would think that whether or not you count twin boundaries in image inalysis procedures would depend somewhat on just what it is you are making the measurements for. If you are primarily concerned with determining kthe phase composition of the material, then I would agree that twins should be ignored; however, if you are trying to correlate your measurements with hardness or strength, then It would seem to me that they should be counted, because they would tend to impede dislocation movement and hence have a strengthening effect. And, Ron is right, trying to do such measurements with a computerized program can be frustrating, largely because it is extremely difficult to get uniform etching. Grain boundaries tend to end up being discontinously etched, and while the eye can make allowance for this, computers have a difficult time doing so.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Does anyone have a great protocol for showing dynein arms in axonemes? Even when the orientation is right, the dynein doesn't seem to show up as well as we would like.
Thanks
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
First, I am assuming the CCD you refer to is likely a GATAN camera. If so, I offer this advice(and it comes with the many voices of experience): Unless the objectives "fall: off the scintillating surface via either CAREFULLY turning the camera on it's side to pour out the trash, OR by CAREFULLY "encouraging" the trash out by applying a slight metered air stream (e.g. try a spray/vacuum bulb such as used on new born babies to relieve nasal blockage) after laying the camera on its side. Once off the surface of the polished scintillator and on the side wall, CAREFULLY using a long-stick swab can remove the trash the rest of the way.
I stressed the word "CAREFULLY" because you don't want to rub the shiny scintillator surface. It scratches as easy as the phosphor on a viewing screen. If the above does not work, send the camera back to GATAN for a scintillator recoating. The cost is about $400 or so and their turn around time is very good. Good luck!
CLay Jordan Customer Service Engineer FEI/Philips Electron Optics
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Hi everybody!
I have a CCD attatched to the bottom of my Philips EM430 that, over the years, has acquired a bit of dust, lost samples, Be rings, etc... I want to be able to clean the surface off before I start my in-situ work, but am a little tentative about doing this. Is there anything I should / shouldn't do if I want to keep my job, i.e. what are the best ways to destroy a CCD from the inside? What would be the best method of going about this? Should I remove the whole camera or is it best to do this inside the column? Thanks in advance for any help.
Brian Gorman bgorman-at-umr.edu Graduate Research Assistant (573) 341-4405 Electronic Materials Applied Research Center 303 Materials Research Center University of Missouri - Rolla Rolla, MO 65409 http://www.umr.edu/~bgorman
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This message is in MIME format. Since your mail reader does not understand this format, some or all of this message may not be legible.
I'm trying to revive an old (mid 50's I'm told) Zeiss UltraPhot2 and am running into the obvious lack-of-parts-problem. I've contacted Zeiss with minimal luck - a few photocopies of old docs and that's it.
Any ideas or leads to parts?
I'm told that the old Universals shared some parts with the UltraPhots.
This is for personal use so there's limited budget (of course!).
I've been following the discussion on various media for archiving and thought I might add another thought. Iomega ZIP drives are gaining a reputation (deserved or not) for a fault that's become known as the "Click of Death". Soon after I read about this, my own 100mb drive (factory installed by Dell) began exhibiting some of the symptoms: repeated rhythmic clicking when trying to access the disk, computer system lock-up for extended periods when trying to use the drive, messages on-screen that the disk is "write-protected" or full, when no write-protection is in force, inability to write to or read from the disk, and general weirdness (drive designations changing from one startup to another, for one thing).
Iomega has admitted this fault, but denies that it's common. Check their info at www.iomega.com. For another perspective, check the site at http://www.thirdeyesp.com/jatin/iomega/#lmformation.
Caution with valuable data is definitely in order. I can no longer reliably access my own ZIP drive.
For what it's worth.
Randy
Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
} A word of caution, be sure that the } cement-workers who do the floors are sober during the process; otherwise, } you're likely to end up with the situation I have in the basement of my } house - there's a lovely drain, but the floor around it has the contour of } a volcano, and so water can't spontaneously reach the drain.
This volcano floor syndrome is not unusual. Rather than lack of sobriety, it is more likely due the difference in density of the floor drain plumbing and the poured concrete. The concrete being much heavier than water will float up the drain (and adjacent cement) unless the drain/plumbing are anchored in place somehow (from below or above).
Bill
} {)))'} {'(((} { Bill Trevarrow, PhD. Zebrafish Facility Director Institute of Neuroscience University of Oregon 1254 Eugene, OR 97403-1254 } {)))'} {'(((} { Off.Tel: (541) 346-4598 Fac. Tel: (541) 346-4512 Fax: (541) 346-4548 e-mail: trevarro-at-uoneuro.uoregon.edu } {)))'} {'(((} {
} I'm trying to revive an old (mid 50's I'm told) Zeiss UltraPhot2 and am } running into the obvious lack-of-parts-problem. } I've contacted Zeiss with minimal luck - a few photocopies of old docs } and that's it. } } Any ideas or leads to parts?
---------- } From: Lincoln, Ian {IAN.LINCOLN-at-kla-tencor.com} } To: 'MSA List' {Microscopy-at-Sparc5.Microscopy.Com} } Subject: LM - Reviving old Zeiss UltraPhot2 } Date: Friday, February 06, 1998 8:16 PM } } Hello all, } } I'm trying to revive an old (mid 50's I'm told) Zeiss UltraPhot2 and am } running into the obvious lack-of-parts-problem. } I've contacted Zeiss with minimal luck - a few photocopies of old docs } and that's it. } } Any ideas or leads to parts? } } I'm told that the old Universals shared some parts with the UltraPhots. } } This is for personal use so there's limited budget (of course!). } } Thanks sincerely for all replies in advance, } } Ian Lincoln } We still carry parts for the older Zeiss equipment. Please contact us at sylviapns-at-worldnet.att.net or call P & S Products at 732-671-5759. Pete Dondl
I adress everybody, who has experience with an XL30 ESEM FEG. I am interested in any kind of user feedback on this instrument, positive or negative. I would be very happy to get information about weak points and shortcomings of that machine in the daily work.
I typically use my Ultracut E microtome with a diamond knife. However, I now have an application that will require me to use glass knives.
I have realized that I do not know how to adjust the height of a glass knife in the knife holder. I am aware of the gauge on the left-hand side, but how does one adjust knife height. The owner's manual merely says that "the height of the cutting edge has to be aligned with the top of the height gauge before fixing the knife with clamping screw".
It would seem to me that merely raising the knife above the surface of the wedge insert would create a less stable knife.
Thank you.
Don ______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
Just to be fair, here is the other side of the story:
update on clicking death Feb 5 98
"Click of death" a matter of math By Paul Festa February 5, 1998, 1:15 p.m. PT news analysis
Problems with Iomega's storage products are emblematic of this category of inexpensive, portable products, according to analysts.
Complaints about both the 100MB Zip drive and the 1GB Jaz drive have spawned Web sites and bulletin boards devoted to anti-Iomega griping. These include the Click Death Home Page, special areas in online publications such as MacInTouch, and voluminous postings in newsgroups including "alt.iomega.zip.jazz" and "comp.sys.ibm.pc.hardware.misc."
But analysts say that the ire directed against Iomega is merely the product of the company's enormous success in selling millions of drives, and also a function of the product's inexpensive construction.
"When you've sold 12 million drives, even a 1 percent failure rate is going to mean a lot of complaints," said International Data Corporation analyst Bob Amatruda. "Ship 12 million and you're a victim of your own success."
Iomega announced last quarter that it had shipped more than 12 million Zip drives since it launched the product in March 1995. Company spokesperson Tyler Thatcher today said that the company sells about 1 million Zip drives every month, so the 12 million figure is already out of date.
Iomega refused to disclose its rate of failure or product return, referring only to the statement it made last week that pegged its customer complaints at below industry norms.
Amatruda said one basic problem with removeable storage was that it was removeable. "People think nothing of sticking them into your shirt pocket or throwing them in the car," he said. "There's a certain level of environmental risk, by the sheer fact of its being removeable."
Disk Trend analyst Jim Porter said the removeable storage technology used by Iomega and its competitors was particularly susceptible to dust contamination because of the microscopic distance between the tape and the drive head.
"The distance from the surface to the disk head is between two and four millionths of an inch," he said. "That's a lot less than any microscopic dust particle. Anything could cause a problem."
Porter stressed that not only were Iomega's products within industry norms for failure, but that the risk for failure was statistically insignificant.
"The risk with removeable media is higher, but it's not even approaching the level that the average user needs to be worried about it," he said. "When you drive the car out of the garage, statistically you're not going to get killed going down the block. But you might."
*****
And here are more details directly from Iomega (issued a couple of days ago):
Iomega today issued a statement saying that "of the more than 12 million Zip drives shipped, Iomega is aware of a small percentage of customer complaints, a number lower than industry norms."
Iomega did not acknowledge the "click of death" problem. The company's Web site, however, has a page that describes it.
Iomega later amended its statement to acknowledge the problem.
"A number of Iomega's customers call from time to time describing a 'clicking' sound emanating from their Zip drive, which can be a symptom of a variety of problems in Zip drives, as well as in other kinds of drive products in general," the statement read. "Iomega continually works with its customers to resolve the particular problems they are experiencing. Iomega also continually evaluates its own product testing data to ensure the highest quality standards."
A source who identified himself as a former Iomega technician said the problem was well-known within the company when he started working there more than two years ago. He said the problem was not common, but noted that it accounted for about half of the malfunctioning drives on which he worked.
The source said the clicking sound is caused by the read/write head bumping against its movement stops--bumpers that keep the head within its intended range--while searching for and not finding track 0 on the Zip disk. When the "click of death" problem happens, the read/write head fails to find that track, which contains vital directory information, because the head has become misaligned.
The cause for that misalignment?
"The drive and disk are not extremely sturdy," the source said. "They're not flimsy, but people like to carry them around, and depending on how your car rides, after six to eight months, you might get the problem."
The source also said that dropping the drive, exposing it to the electromagnetism of a computer monitor, and other external factors could cause the misalignment. He noted that internal drives were less susceptible to the problem, and stressed that, apart from the sturdiness of the casing, the products themselves were not defective.
"I don't think the drives are faulty in any way," he said, noting that he had just authorized the purchase of 50 Zip drives for the company where he is currently employed.
The source said he also had encountered a related problem reported in newsgroups: a domino effect in which misaligned heads damage disks, which in turn misalign heads of other drives, which then damage more disks.
"It's fairly rare," he said. "But it does happen." ********
We have more than 20 Zips in my group, which have been in use for more than 2 years, and as far as I am aware, we have had no failures. I wonder if the drives died quietly, like many of the hard drives or floppy drives we have had fail in the same period of time, would this be a big issue? :-)
Larry
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Dr. Allard's response to my recent posting about ZIP drives gives a valuable perspective on this problem, if indeed it is a problem at all.
To clarify a couple of points, the drive on my computer is internal, factory-installed in a mini-tower and located about four feet away from the monitor. It has seen very little use in the three months I've owned this system and has never been subjected to shock. On the other hand, the disk which was in use when the problems originally showed up had been, I discovered, carried in a briefcase without its own protective case, on at least one occasion. Now the drive acts up with any disk. This may be relevant to Dr. Allard's information.
I have also read Iomega's web page and other material, and I found a couple things a little odd. The repeated statement or implication that the drives are "not extremely sturdy" juxtaposes strangely with the statement that "I don't think the drives are faulty in any way". If they are indeed so delicate and prone to dust contamination, I can only repeat that trusting valuable or irreplaceable data to them should be a matter for caution and backups. (Of course, backups should always be made regardless of the medium used.) Apparently the days of rugged 3 1/2" and 5 1/4" (remember them?) drives and disks are past. Mine still function flawlessly from a decade ago, but....
I agree with Dr. Allard that many users of these drives have had ZERO problems with them. Check out the options and buy what suits your needs, but be aware of what others have experienced, good and bad.
All the best,
Randy
Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
In the accessories-box of your Ultracut E there should be (or have been when it was new) a small metal plate that fits in the knifeholder's botto= m and acts as a raiser, so that you can use standard glass knives. If this plate has been lost, you might take one from another ultramicrotome, measure its thickness and make (or have one made) that fi= ts your particular knife holder. On the other hand, there is of course the option to get one from Leica.
Second, I too was a bit surprised at the implications that Zips should be handled a bit gently. In my experience, I have chucked drives into suitcases or shipping boxes or briefcases etc and carted them all over the world (well, at least once to Japan), and I routinely carry a half dozen disks, unprotected by their jewel cases, in my computer briefcase. I have never had a problem with drives or disks. In fact, on one occasion shortly after I got my first external Zip drive a couple of years ago (still in use in my home office), I was transferring data when my son knocked the drive off the top of my stack of peripherals and I found it hanging by the cable, still merrily dumping data without a hitch. So my impression has been from the beginning that the drives and disks are basically bulletproof. But now I may just be a bit less cavalier in handling the disks, given that they contain a lot of data, and are certainly susceptible to dust contamination, as are any similar floppy-type devices. (It would be nice to have some plastic envelopes for the disks, rather than those bulky jewel cases, don't you think?)
But I still think that Zips are the best thing going for the purposes they satisfy, and I am not planning to discard all of the drives and disks we presently use at my lab if I ever have one fail on me... I'm sure our users would revolt, since they universally depend heavily on Zips to take their images home with them.
Just MHO.
Larry PS you can always try the famous "throw test"...take a Zip disk and sail it down the hallway 40-50 feet, then stick it in your drive and see if it works. I'll bet it does, and I also bet you won't be as lucky with any other type of high density removable storage...
PPS As is well-known, I have held shares in Iomega for several years, as well as in several other drive manufacturers. But all of the above details are true anyway.... :-).
} Dr. Allard's response to my recent posting about ZIP drives gives a valuable } perspective on this problem, if indeed it is a problem at all. } } To clarify a couple of points, the drive on my computer is internal, } factory-installed in a mini-tower and located about four feet away from the } monitor. It has seen very little use in the three months I've owned this } system and has never been subjected to shock. On the other hand, the disk } which was in use when the problems originally showed up had been, I } discovered, carried in a briefcase without its own protective case, on at } least one occasion. Now the drive acts up with any disk. This may be } relevant to Dr. Allard's information. } } I have also read Iomega's web page and other material, and I found a couple } things a little odd. The repeated statement or implication that the drives } are "not extremely sturdy" juxtaposes strangely with the statement that "I } don't think the drives are faulty in any way". If they are indeed so } delicate and prone to dust contamination, I can only repeat that trusting } valuable or irreplaceable data to them should be a matter for caution and } backups. (Of course, backups should always be made regardless of the medium } used.) Apparently the days of rugged 3 1/2" and 5 1/4" (remember them?) } drives and disks are past. Mine still function flawlessly from a decade } ago, but.... } } I agree with Dr. Allard that many users of these drives have had ZERO } problems with them. Check out the options and buy what suits your needs, } but be aware of what others have experienced, good and bad. } } All the best, } } Randy } } } Randy Tindall } 2017 Princess Jeanne } Las Cruces, New Mexico 88001-4157 } } rtindell-at-nmsu.edu (work) } nrtindall-at-zianet.com (home)
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
It is interesting to hear about sudden "out of the sky" problems with ZIP removables. You may want to check my odyssey notes of the SyQuest drives below, to rule out similarities to your ZIP situation.
As I had heaps of trouble with SyQuest 44 MB Drives, which were quite occasional in nature, but devastating in effect, I chased down some common denominators for SyQuest related problems.
I did not find any SyQuest 44 MB drive, which ever broke, although they SEEMED broke at times. It is the **circumstances** where they are hooked up to, which make them work or not.
(1) If the external case, where the SyQuest sits in, has had it, you would not get an alert. Instead, your disks re-re-mount, keep scratching, or just stop in the middle of a disk access. They even might freeze the computer for some time. Only remedy is, to get a new case with new power supply. You also can screw the device directly into a minitower, such as I did into a Macintosh Quadra 800 (regular SCSI bus) - to immediately stop the problem, or into another minitower such as Macintosh Quadra 840AV (one of the fastest SCSI buses ever) - to continue screwing up, I caused also an internal SCSI cable fire (but was able to extinguish within minutes).
The same holds true for CD-recorders. They operate perfectly on one SCSI chain, but they just do not work on others.
We thought also, Martin's SyQuest has gone broke. But then I ripped it out of the external case and shoved it directly into a Macintosh II something (yes, we had to saw something off the case), and it keeps working more than perfect upt to date as it never did before.
SCSI chain problems can be aggravated or alleviated by anything such as different cables, different harddisks etc. Some new harddisks such as certain Quantum Fireballs at one stage were thought by some people to have a bug, only until FWB released the support file for their RAID toolkit for this particular Harddisk, and now everything is perfect.
(2) If the controller gets slightly too hot, it stops working. Cooling seems to be very important. I know someone who lost a whole harddisk simply because he failed to regularly check the FAN of the external harddisk case and give it some oil. Internal disk slots may be properly ventilated (remember the additional fan you could get in case you wanted to put two drives in the lowest slot in a Quadra 800 ?) - but they may not be (cheap mini towers).
(3) There is such a thing as bad media. Once dropped, once next to a power cable, bye bye data. That is why I reformat them when they come back by mail and I managed to retrieve all data.
CD - burning and having multiple external (and internal) harddisks for on-the-fly and hot-start-system-restore setup has taken these time consuming problems away from me up to some extent. I would never put anything on a removable medium, which is not backupped on at least two other removable media.
My thoughts on
--------------------------------------------------------------------------- Wolf Schweitzer MD mailto:wschweitzer-at-access.ch
The biggest problem with the Zip-like drives (and here I am including *all* these sons and daughters of the Winchester) is that they are all transitional technologies and that there is little real knowledge about their obsolescence cycle. They are simply not archival devices.
That means that they may be great for temporary storage of data -- for transferring data from one machine to another, etc. -- but they are probably not (in my opinion) the best thing for archving or for keeping your *only* copy of data.
The thing that many folk getting into digital imaging forget is that sticking data on a disk (or tape) is not a fire-and-forget strategy for keeping data.
There are three things to worry about:
1) Media degradation 2) Media obsolescence 3) Format obsolescence
Different media have different half-lives. Older 9-track tapes, for instance, often had errors as early as 5 years into their life cycles, and typical obsolescence was about 7 years. Newer tapes are better, and Exabyte estimates a 30-year lifetime for its 8mm tapes ***if*** you don't use them much (e.g. a tape used for a daily backup will not last anywhere near that).
Disks, including Winchester variants, tend to have a much shorter life span. That means that if you are saving stuff to these disks as archival, you will need to recopy the data in a couple of years.
As a case in point, I used to archive data on 88MByte Syquest disks. I have one disk written in 1993 in which only about 40% of the images are readable without error, another 30% are partially readable with errors in the image, and the rest are beyond useful salvaging.
But more important, remember that in 10 years nobody will be using Zip (or Syquest or whatever) drives, and you may not be able to find a machine that will even be able to read your data. Eight months ago, for instance, we got rid of our last 9-track tape drive in my lab. We had approximately 300 9-track tapes full of data, and we can no longer get to it without going somewhere else.
How many new computers are sold today with 5 1/4" floppy drives? When was the last time you saw an 8-inch floppy drive? An 11-inch drive? A paper-tape machine? A card-reader? Or, more to the point, 10MByte Bernoullis (which were also made by Iomega), 20MByte Bernoullis, 22 MByte Syquests, etc.?
In 20 years, CD-ROM players will be as common as 8-track players in new cars.
Finally, in 20 years, many of the formats you are storing images on will no longer be supported. How many of your applications can read ILBM and other Amiga format files? /usr/image files? Over the next few years, common formats, including .gif and others will become either vanishingly rare or astonishingly mutated. Remember that neither GIF nore JPEG formats are "standard" for the net -- that has gone to PNG. There are early TIFF files around that many new applications cannot read because of the nonstandardization of compression in "standard" TIFF. Those of you who know UNIX freeware know that the famous libtiff library supports only a subset of TIFF variants.
All of this is to say that the occasional error in a Zip disk is pretty much to be expected, not only because of the magic of low error rates in large sales volumes, but also because it is not *meant* to be an archival or permanent media. It is a transitional media for temporary storage of data. As such, they are great.
If you are moving data from one machine to another, if you are looking for a place to park data with moderate turnover, etc. they are tremendous. However, hey are not archival devices, and I'm not sure it's a good idea to have your only copy of important data on one of these things if you are looking for a sole or long-term storage solution.
billo
On Sat, 7 Feb 1998, Randy Tindall wrote:
} } Dr. Allard's response to my recent posting about ZIP drives gives a valuable } perspective on this problem, if indeed it is a problem at all....
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Thanks to all who responded to my request to DTSA spectra. I = unfortunately had a 40 ft tree fall at my home, tearing out gas and water = lines and several other things, SO I will try the spectra sent and = respond to other questions, but it will take me a couple of weeks to get = to it because of this unscheduled emergency.
Also, for those who asked what format to send, the MSA format is OK, as = it can be read by DTSA. Thanks Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
(SMTPD32-4.03) id A0912BAD0032; Sun, 08 Feb 1998 18:32:33 EST Message-ID: {34DE40A0.4C4C-at-map.com}
Reminder: The deadline for early registration is rapidly approaching!
"Optimizing Light Microscopy" A lively lecture-demonstration on Light Microscopy offering a wide range of tips and techniques from set-up to trouble shooting, contrast enhancement to video microscopy. March 16 - New York City - The Beacon Hotel March 18 - Springfield, MA - The Radisson/ West Springfield March 20 - Boston, MA - Tufts Medical School/Multi-Media Resource Center
Tuition includes a copy of the newly released book Optimizing Light Microscopy. Discounts are available for early registration or multiple registration from the same lab.
For further information, contact Barbara Foster or Ken Piel at Microscopy/Microscopy Education: (413)746-6931 or respond by email: mme-at-map.com.
Does anyone know of any sources for having basic SEM analysis done without EDX capabilities? Are the images available in a digital format for distribution? I would also like to know the hourly rates and turn around time if possible.
} Date: Sun, 8 Feb 1998 18:43:40 +1000 } To: microscopy-at-Sparc5.Microscopy.Com } From: Wolf Schweitzer {wschweitzer-at-access.ch} } Subject: Re: Possible cautions about ZIP drives } Cc: } Bcc: } X-Attachments: } } It is interesting to hear about sudden "out of the sky" problems with ZIP } removables. } You may want to check my odyssey notes of the SyQuest drives below, to } rule out } similarities to your ZIP situation. } } } As I had heaps of trouble with SyQuest 44 MB Drives, which were quite } occasional in } nature, but devastating in effect, I chased down some common denominators } for SyQuest } related problems. } } I did not find any SyQuest 44 MB drive, which ever broke, although they } SEEMED broke } at times. It is the **circumstances** where they are hooked up to, which } make them work or not. } } } (1) If the external case, where the SyQuest sits in, has had it, you would } not get an alert. } Instead, your disks re-re-mount, keep scratching, or just stop in the } middle of a disk access. } They even might freeze the computer for some time. Only remedy is, to get } a new case with } new power supply. You also can screw the device directly into a minitower, } such as I did into } a Macintosh Quadra 800 (regular SCSI bus) - to immediately stop the } problem, or into another } minitower such as Macintosh Quadra 840AV (one of the fastest SCSI buses } ever) - to continue } screwing up, I caused also an internal SCSI cable fire (but was able to } extinguish within minutes). } } The same holds true for CD-recorders. They operate perfectly on one SCSI } chain, but they } just do not work on others. } } We thought also, Martin's SyQuest has gone broke. But then I ripped it out } of the external case } and shoved it directly into a Macintosh II something (yes, we had to saw } something off the case), } and it keeps working more than perfect upt to date as it never did before. } } SCSI chain problems can be aggravated or alleviated by anything such as } different cables, } different harddisks etc. Some new harddisks such as certain Quantum } Fireballs at one stage were } thought by some people to have a bug, only until FWB released the support } file for their RAID } toolkit for this particular Harddisk, and now everything is perfect. } } } (2) If the controller gets slightly too hot, it stops working. Cooling } seems to be very important. I know } someone who lost a whole harddisk simply because he failed to regularly } check the FAN of the } external harddisk case and give it some oil. Internal disk slots may be } properly ventilated } (remember the additional fan you could get in case you wanted to put two } drives in the lowest slot } in a Quadra 800 ?) - but they may not be (cheap mini towers). } } (3) There is such a thing as bad media. Once dropped, once next to a power } cable, } bye bye data. That is why I reformat them when they come back by mail and } I managed to retrieve all data. } } } CD - burning and having multiple external (and internal) harddisks for } on-the-fly and hot-start-system-restore setup has taken these time } consuming problems away from me up to some } extent. I would never put anything on a removable medium, which is not } backupped on at least two } other removable media. } } My thoughts on } }
--------------------------------------------------------------------------- Wolf Schweitzer MD mailto:wschweitzer-at-access.ch
I am sure that you can delete this message right now, if you have never ever used Link ISIS Suite Revision 3.2.
I got a few "?" about the basic functions of the software as following:
When I try to download a spectrum as a TIFF file into a floppy disk (select File/Export as TIFF (Solid Spectrum)), the machine does behave by herself, but the file saved is as big as over 300k. It would be only about 10k if the same spectrum would be captured by using a software such as Paint Shop Pro (PSP). Why?
If you think that this is not a problem at all, and the advantage to use the built-in function is to save your time, please be careful!!! You may loss your important data as there is no warning notes on the screen when the floppy disk is over-flow! --- 10's file names would be on the list but only a couples of them has been saved.
The similar problem existed when you try to save Linescan results. You can do it by selecting Print/File/Save Results as TIFF. The file saved in this way could be as big as over 200k, compared to only about 10k captured with PSP. The another problem with this function is that you must pull the print preview window away from the sight of "camera" --- do not cover the linescan group windows which you want to save. Why? Why not?
{P} I am sure that you can delete this message right now, if you have never ever used {U} Link ISIS Suite Revision 3.2 {/U} .
{P} I got a few "?" about the basic functions of the software as following:
{P} When I try to download a spectrum as a TIFF file into a floppy disk (select File/Export as TIFF (Solid Spectrum)), the machine does behave by herself, but the file saved is as big as over 300k. It would be only about 10k if the same spectrum would be captured by using a software such as Paint Shop Pro (PSP). Why?
{P} If you think that this is not a problem at all, and the advantage to use the built-in function is to save your time, please be careful!!! You may loss your important data as there is no warning notes on the screen when the floppy disk is over-flow! --- 10's file names would be on the list but only a couples of them has been saved.
{P} The similar problem existed when you try to save Linescan results. You can do it by selecting Print/File/Save Results as TIFF. The file saved in this way could be as big as over 200k, compared to only about 10k captured with PSP. The another problem with this function is that you must pull the print preview window away from the sight of "camera" --- do not cover the linescan group windows which you want to save. Why? Why not?
I am looking for an image analysis for a research department. It should be based on a Windows PC, easy to handle and programmable by a macro language. It should be easely addapt to different problems. It should have different algorithem for detection/segmentation as well as the separation of particles. The image analysis will be used for light microscopy as well as for SEM and TEM.
Are there any papers, where image anaysis programs are compared?
Which is a good listserver for image analysis?
Are there any suggestions for a good image analysis program?
I am actually working with semithin sections of SPURR embedded microarthropods after Formaldehyde-Cetyl Pyridinium Chloride fixation. These sections shall be stained with a mixture of Toluidine-Blue/Methylene-Blue/Sodiumtetraborate to give sharp results. The problem we have is that this staining is rapidly fading if the slides are mounted with media like Entellan or Eukitt, probably due to the Xylene content. This might be also a problem of SPURR because with other resins like LR-White or GMA this doesn't happen. My attemps to use SPURR as a mounting media, followed by heating at 70 degrees overnight, were not very sufficient because the resin remained sticky for unknown reasons.
Any comments, eg. about alternative mounting media, are welcome.
Jens
--------------------------------------------------------------- Dr. Jens Buecking Tel. +49-(0)421-218 3745 University of Bremen Fax. +49-(0)421-218 4620 Dep. of Biology Email jbueck-at-biologie.uni-bremen.de Leobener Str. - NW2 28359 Bremen ---------------------------------------------------------------
Ian, I have an Ultraphot III, still the best optical microscope we have on site (Incredibly versatile - you can do anything! And beautifully engineered). I would also be interested to hear of any parts / accesories which may be around. But no, you can't have any of my bits. I like the machine too much.
Regards,
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK
} Hello all, } } I'm trying to revive an old (mid 50's I'm told) Zeiss UltraPhot2 and am } running into the obvious lack-of-parts-problem. } I've contacted Zeiss with minimal luck - a few photocopies of old docs } and that's it. } } Any ideas or leads to parts? } } I'm told that the old Universals shared some parts with the UltraPhots. } } This is for personal use so there's limited budget (of course!). } } Thanks sincerely for all replies in advance, } } Ian Lincoln
Ron, Try Anatech, Ltd. in Springfield, VA (1-800-752-7629) or (703-941-8860). I don't know who else supports Hummer products and I'm obviously not endorsing any one vendor.
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124
} ---------- } From: Hybertson, Ron[SMTP:ron_hybertson-at-ms1.mankato.msus.edu] } Sent: Friday, February 06, 1998 8:39 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Hummer Info } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } I am looking for a vendor of the HUMMER V sputter coater electrode. } } } } }
I need the post adress Mrs E.Weck or Mrs E.Leistner (from Munchen ?), They are author's; Metallographische Anleitung zum Farbatzen nach dem Tauchverfahren Teil I, II and III edited in Dusseldorf - DVS 1983
best regards for all
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
Small World introduces a new one day EDS course. The course gives you the practical knowledge you need to run better samples. If you don't have time to attend the week long courses, but would like to become a better analyst, this is the course for you. Check out our web site for more information:
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Drosophila/Resin 2/9/98 10:32 = AM
Dear Microscopists: I am looking for a protocol for the embedding of drosophila embryos in = either epoxy resin or spurr resin. Would anyone be able to help? Thanks = in advance. Linda Chicoine Center for Cell Imaging Yale University New Haven, CT USA 203-785-3646 phone 203-785-7226 fax
I am starting a study in which the size and distribution (may be shape) of fat globules in milk samples can be of relevance. I wonder if I can visualize these fat globules by LM (and if so, how, directly after some staining) or if other techniques (I might think of SEM with cryofracture) are appropriate.
I would be very grateful to any advice I receive.
Yours,
Antonio Molina-Garcia, Inst. del Frio, CSIC Madrid, SPAIN Dr. P.D. Sanz Engineering Department Instituto del Fr=EDo (C.S.I.C.) Ciudad Universitaria Calle Ramiro de Maeztu, s/n 28040-Madrid E-Spain
You have hit the nail directly on the head...I could not have said it better. As the technology develops, there will be a necessity to constantly transfer onto up-to-date storage media important data that you have archived. That is probably one advantage of film...it lasts a long time, and would be able to be re-printed, or simply scanned and digitally output, for the forseeable future. But I certainly do not expect that in the year 2010 it is a guarantee that all my lab's CD-ROMs filled with TEM and SEM images will be able to be read out on anything but some clunker CD player we have kept around for that purpose. Of course, by that time I will be retired, and probably won't give a hoot... ;-).
Larry
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
} I have realized that I do not know how to adjust the height of a glass } knife in the knife holder.
Don: Our standard procedure is to focus the binocs in the middle of the gauge, then bring the edge of the knife into focus by lowering or raising. Hope this helps, and this height adjustment is probably more related to the cutting arc than to knife stability.
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
I'm trying to put together a protocol for a user that wants to embed (for TEM) cultured cells without removing them from the surface that they are attached to. I understand that certain kinds of plastic dishes and coverslips tolerate being processed for thin sectioning and can even be sectioned. Could someone enlighten me on the types of plastic involved (PERMANOX ?) and where I might find these items? I'm more interested in coverslips than petri dishes, but I'm open to being influenced otherwise.
I've checked the MSA listserv archives (Thanks Nestor) and several vendor sites, but I still don't have a clear answer. Vendors are certainly welcome to respond.
Perhaps if we keep this off-list, I can just post a summary of the responses.
Thanks, Doug ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
Hi, Are there any users out there using 800 - 1000 degrees heating stages in a SEM?
We would like to look at phase transformations in metal alloys. We also project to buy a new SEM. One of the questions is: Is the secondary electron contrast not too low in such conditions (will it be possible to detect something at high temperature?)
Any experience in this domain would be greatly appreciated. Many thanks Monique repoux-at-cemef.cma.fr
----------------------------------------------------------------------- Monique Repoux Tel: 33 (0)4 93 95 74 13 CEMEF 33 (0)4 93 95 75 91 Ecole des Mines de Paris B.P. 207 Fax: 33 (0)4 93 65 43 04 06904 SOPHIA ANTIPOLIS CEDEX e-mail: repoux-at-cemef.cma.fr FRANCE -----------------------------------------------------------------------
I work in an EM Lab in Minneapolis, MN. Our work is primarily clinical--EM done on biosies of renal and neuromuscular tissue as well as some tumor cases. My question is this: Does anyone have an idea of approximately how many EM Labs are out there (primarily in the US) that function primarily as a clinical laboratory like ours?? I know there are a great deal of research labs tied in with universities, but I am specifically looking for clinical labs. If any one has any information, they can contact me by email at: hcmcemlab-at-sprintmail.com
Donald- there is (or was) an insert (a wedge shaped one on those machines) which set the appropriate knife height assuming you're making "standard" knives -Mike
On Sat, 7 Feb 1998, Donald Lovett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } } I typically use my Ultracut E microtome with a diamond knife. However, I } now have an application that will require me to use glass knives. } } I have realized that I do not know how to adjust the height of a glass } knife in the knife holder. I am aware of the gauge on the left-hand side, } but how does one adjust knife height. The owner's manual merely says that } "the height of the cutting edge has to be aligned with the top of the } height gauge before fixing the knife with clamping screw". } } It would seem to me that merely raising the knife above the surface of the } wedge insert would create a less stable knife. } } Thank you. } } Don } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } P.O. Box 7718 fax: (609) 637-5118 } The College of New Jersey } Ewing, NJ 08628-0718 } } } }
This is a nice question and very often not understood. Putting it basically, with frame averaging the SEM scans the same frame = ( whole picture ) a number of times and stores these images in the same = location in memory. So as the information of you sample is present in = each picture this will become very prominent. All the noise that occurs = in the image is very erratic and so with successive frames will = disappear with reference to the actual image. Frame averaging is like taking a number of course print transparencies = of the same image and lying them one on top of each other. So when = operating in this mode you can select how many images you would like to = store one on top of another. If you select say three images as the third = image is completed the fourth will then replace the first, the fifth = will replace the second and so on. The more images you have piled on top = of each other the clearer the image will appear. However if you now = change the image by moving the stage or the mage, then it will take that = number of frames selected before the change is fully visible. With line scan the SEM will scan the same line a number of times each = time averaging out the noise from the true info to ensure a clear image. If you have a delicate sample then frame averaging is the only choice. = Here the energy of the beam is scanned over the whole area of the = sample. In line scan mode, scanning the beam repeatedly over the same = line can damage the sample. If you are using small spot sizes or low kV operations on a SEM you will = find that the image is very noisy. To then use a bit of frame averaging = makes it easier to see what is happening. Typically on a LEO S 360 we = would use Frame ave of 2 and then a scan rate of TV/8. In this way you have a fairly quick scan rate to reduce sample damage = and still enough response on the change of the image by selecting only a = 2 frame ave, rather than say a 8 or 10 frame ave. If sample damage is not a problem, line average will give the clearest = picture as this replicates what the photo crt does to give you a good = micrograph.
I hope that explains it a bit for you.=20 I am sure there are going to be much more detailed explanations from far = more experiences operators than myself. Best advice I can give you is try it and see what happens. =20 Luc Harmsen Anaspec, South Africa Technical support for E.M. operators, world wide. anaspec-at-icon.co.za TEL: ++ 27 (0) 11 476 3455 FAX: ++ 27 (0) 11 476 7290=20
----------
Hi =20 COuld anyone explain the difference and advantages of these two=20 scanning mode in the SEM? =20 Thanks F
At 03:38 PM 2/9/98 +1100, you wrote: } Hi, Netmates, } } I am sure that you can delete this message right now, if you have } never ever used Link ISIS Suite Revision 3.2. } } I got a few "?" about the basic functions of the software as } following: } } When I try to download a spectrum as a TIFF file into a floppy disk } (select File/Export as TIFF (Solid Spectrum)), the machine does behave } by herself, but the file saved is as big as over 300k. It would be only } about 10k if the same spectrum would be captured by using a software } such as Paint Shop Pro (PSP). Why? } } If you think that this is not a problem at all, and the advantage to } use the built-in function is to save your time, please be careful!!! You } may loss your important data as there is no warning notes on the screen } when the floppy disk is over-flow! --- 10's file names would be on the } list but only a couples of them has been saved. } } The similar problem existed when you try to save Linescan results. } You can do it by selecting Print/File/Save Results as TIFF. The file } saved in this way could be as big as over 200k, compared to only about } 10k captured with PSP. The another problem with this function is that } you must pull the print preview window away from the sight of "camera" } --- do not cover the linescan group windows which you want to save. Why? } Why not?
I only have version 3.1 so I cannot comment on saving spectra, but I can comment some on the Linescan issue.
ISIS saves the results as DIB (device indepent bitmap) files which are effectively the same as Windows BMP files. When I saved a 6-field linescan group, it was stored as a 24-bit color image 784x280 pixels and required 684 Kb. That is 784x280 pixels times 3 bytes per pixel = 660 KB plus some extra header info for good measure. If I resave the image to a different format, I got the size down to 76 Kb as a PCX file or 44 Kb as a JPG file (not recommended).
In other words, it is pretty much as you should expect. I don't know if ISIS always saves in 24-bit true color mode (my monitor was set up as 16-bit color at the time). But DIB does not do image compression of any kind. TIF, GIF, and PCX do compression and can really knock the size down when the images have large areas of the same shade. JPG does compression too, but at the expense of fine details. I would not suggest it for line are or spectra. It is good for pictures. Your PSP probably uses image compression as a matter of course.
As far as making sure the print preview window must be moved, I think it is a software bug. I am not sure if it is on Microsoft's or Oxford's side, but I guess it is a feature of MS. As far as getting no error when the floppy fills up, I don't know about that one. Might be another MS "feature". I would rather save to hard disk for sake of speed, and then use a file manager to move the files to a floppy.
Hope this helps. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
At 12:42 PM 2/9/98 +0000, you wrote: } Hi } } COuld anyone explain the difference and advantages of these two } scanning mode in the SEM? } } Thanks } F
I am much more familiar with the distinction between point averaging and frame averaging.
Point "averaging" has to do with multiple measurements (or lengthened measurements) per point during a single frame scan. For example, instead of dwelling for 25 us per raster point, the dwell might be increased to 100 us or 4 successive readings might be averaged.
Frame averaging has to do with making multiple raster scans over the same area and summing and averaging the readings at each point. For the previous example, the whole frame would be scanned 4 times and averaged point-by-point.
Line averaging might be a similar thing dealing with multiple passes on a line before moving on to the next line.
The problem with frame averaging can be drift between successive frames for long frame times. It would result in blurry images. The drift would still be there for point or line averages but might not be noticeable. The drawback with point averaging would be the increased electron dose per unit of time. There would not be time to recover before the next measurement. That may not be a problem for many samples, but might be important to others. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
For most applications standard embedding protocols can be used with drosophila tissues. I typically use EmBed resin. Dehydrate with ethanol using 20--30 minute soaks, infiltrate with resin:propylene oxide 1:1 for 2 hours-overnight, fresh epoxy resin for 2 hours-overnight, fresh resin, polymerize at 60C for 48 hours. I find that 1.5 ml microcentrifuge tubes work well as processing containers. If orientation is important (usually) then I spread the embryos out on a flexible mold and use minimal resin. It should be about the thickness of a coverslip ~.1-.1mm. I cut out the desired embryo after only 24 hours of polymerization and mount it on a block (previously made with Beem capsules) with a small drop of epoxy and polymerize for another 24 hours. Wooden applicator sticks sharpened to a small flat tip work well to manipulate the embryos in the unpolymerized epoxy. Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
How fresh should the working dilution of 2% buffered glutaraldehyde be for diagnostic TEM? Our biopsy material is infrequent, and we don't want to make up quantities that would end up being discarded before viable use. Thanks Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children Dallas
Dear Jens, this problem arises very often and I have it seen with Epon, Spurr's as = well as also Lowicryl. What the exact mechanisms is, I can't tell you ( = I think/thought about the mounting medium itself, the xylene/toluene, = also the resin(s) itself, weak binding of the -normally- highly alkaline = staining solutions like your toluidineblue/methylene blue variant in = sodium-borate buffer: I guess, about pH 9-10, or??) I wrote "thought about it" because I don't have any problem with this = sort of "leaching out of staining" in mounted semithin sections (which I = noted sometimes starting immediately after having had sections mounted, = i.e. within 2-3 days or so, as well as after 1-2 years: no staining of = the sections at all, but veils of blue and red over the whole cover slip = area).
My suggestion: If you don't need to mount your sections permanently, try = the following: DON'T mount your sections at all: save EUKITT/ENTELLAN, SAVE TIME and = MONEY: Stain your sections as usual, be happy, if you get brilliant = staining. If you want to *look only* to your stained sections (e.g. for = correlative TEM: searching for locations/areas for subsequent = ultrathins) your sections readily show the details wanted without a = cover slip and mounting. If you want to PHOTOGRAPH your SECTIONS: place small drops of immersion oil onto your sections, cover with a = cover slip as usual (taking care of air bubble formation) for viewing = with normal objectives (e.g. x4, x10, x25, etc., provided correct = Koehler's illumination). Best results with immersion oil objectives (e.g. x50, x60, x100) with = drops of immersion oil onto the sections, WITHOUT using a cover slip = (directly).
How to store those sections then: get rid of the cover slip (slipping = away the cover glass, immerse object slide into xylene or toluene bath = (2 x 2-5 min), preferably in a coplin jar with lid; then wipe gently = with soft, clean cloth and or dry the slide by means of compressed air = (pressured air, or something like "DUST OFF".....).
You can store your slides then without any problem: you will have = brilliant staining after 5 years too, if stored in (a) light-tight and = dustfree box(es). If it happens after long time storage (what sometimes appears to happen) = that your staining is not any more that brilliant as you expected, then = only repeat your staining once more again........(no need of de-mounting = your sections etc.....)
This technique/possibility (unfortunately *not patented*) I am dealing = since now 15 years of diagnostic correlative LM/TEM-practice (all of the = semithins will be stored for later comparison if needed....) without any = problem .
Hope this helps, best regards
Dr. Wolfgang MUSS , just finishing a long working day, Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: Jens Buecking[SMTP:jbueck-at-biologie.uni-bremen.de] Gesendet: Montag, 09. Februar 1998 13:44 An: Microscopy-at-sparc5.microscopy.com Betreff: LM - fading of stained SPURR sections
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Dear all,
I am actually working with semithin sections of SPURR embedded microarthropods after Formaldehyde-Cetyl Pyridinium Chloride fixation. These sections shall be stained with a mixture of Toluidine-Blue/Methylene-Blue/Sodiumtetraborate to give sharp results. The problem we have is that this staining is rapidly fading if the = slides are mounted with media like Entellan or Eukitt, probably due to the = Xylene content. This might be also a problem of SPURR because with other resins like LR-White or GMA this doesn't happen. My attemps to use SPURR as a mounting media, followed by heating at 70 degrees overnight, were not = very sufficient because the resin remained sticky for unknown reasons.
Any comments, eg. about alternative mounting media, are welcome.
Jens
--------------------------------------------------------------- Dr. Jens Buecking Tel. +49-(0)421-218 3745 University of Bremen Fax. +49-(0)421-218 4620 Dep. of Biology Email jbueck-at-biologie.uni-bremen.de Leobener Str. - NW2 28359 Bremen ---------------------------------------------------------------
Assuming you have some standards for the line and it is at a decent energy, should it really make any difference whether a K, L, or M line was used? The excitation is already done, and isn't a 3 keV x-ray behave the same regardless of how it was generated?
I know that we get in trouble if we try to do "standardless" analysis on one of our systems with an L-series line. Somehow the database seems to account for K and M lines well enough, but seems to have a systematic problem with L lines. Therefore, we standardize.
At 07:58 AM 2/6/98 GMT+2, you wrote: } I am analysing bulk alloys of high atomic number elements eg. Re, Ir } (Z=75, 77) in a light element matrix eg Al (Z=13) at 20 kV with an } EDS system in an SEM using ZAF corrections and pure element } standards. Typically, the high Z elements have L alpha peaks at } around 9kV, M alpha peak at around 2kV. The high Z content ranges } from 0.1 to 50 at%. } } As I understand it, correction factors for L lines are better than } for M lines. However, the M peak is much higher than the L peak at } 20kV, so statistics are better. At the low concentrations of high Z } element, I only see an M peak, not L peak. There is about up to 2 } at% difference analysing using both peaks over the alloy composition } ranges and such a difference is a problem for our work. } Unfortunately I do not have any alloy standard to check results } against! So which peak to use? Yes, someone analysed on } a microprobe (using WDS) utilising same area on a sample and same } standards and got a third answer which gave no direction to my } question! } } Any advise would be gratefully received. Thanks in advance. } Mike ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
At 12:42 9/02/98 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America We have a Leica/Cambridge S-360 on which you can choose either line or frame averaging to achieve the same exposure or amount of integration (noise reduction).
I usually use line averaging - around 40-50 scans per line depending onthe signal strength. Line averaging conceals any image drift if the specimen is somehow moving.
BUT frame averaging is very good for reducing charging aretefact. The dwell time per point can be made fast enough that the chage deposited is small and leaks away between frames.
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Ok, I know it is a little early for most of you to be considering the Microscopy and Microanalysis Meeting in 1999 (in Portland Oregon, by the way), but I want to try and get some feedback on the subject of physical science tutorials.
I am in charge of organizing the physical science tutorials and I am seeking input from the microscopy and microprobe community.
What would you like to see in the way of tutorials at the 1999 meeting?
Please make suggestions directly to me and I will summarize to the net if the need arises.
In 1997 we had tutorials on environmental scanning electron microscopy (Stuart McKernan, Brendon Griffin and Chris Gilpin) and electron back scattering pattern acquistion and analysis (Joe Michael).
In 1998 we will have specimen preparation (tripod polishing, microtoming of inorganic materials will be highlighted). This will be "The Ron and Tom Show" starring Ron Anderson and Tom Malis.
Further in the past there have been tutorials on a wide variety of subjects. Some of the most popular were those related to computer acquistion and processing of images. In fact anything computer related seems to be popular. Is there still interest here or is everyone now comfortable with their computer systems?
Please let me know by replying to this email or by calling the number below.
Thanks.
Cheers
Jfm.
________________________ Note new Area Code (734) ________________________ John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Hi Jens, I have a suggestion for increasing the intensity of staining in hydrophobic resins (eg Spurr's). Your stain solution should be made up at high pH so that protons in solution don't compete with stain molecules for binding sites (recipe below). The metachromicity of toluidine blue depends on water molecules being present and so mounting media like Entellen (Xylene base?) ruin this effect. My trick is to exhale fairly forcefully onto the sections just before adding a solvent-based mounting medium. This adds just enough water so that colours appear as desired. If you do enough slides at once the hyperventilation combined with the effect of breathing mountant fumes gives one a special feeling all over. I have not noticed fading in slides prepared this way.
0.5% toluidine blue in 0.1% sodium carbonate (pH 11.1).
You should be able to make up other stain solutions to high pH in a similar fashion. Cheers, John
} Dear all, } } I am actually working with semithin sections of SPURR embedded } microarthropods after Formaldehyde-Cetyl Pyridinium Chloride fixation. } These sections shall be stained with a mixture of } Toluidine-Blue/Methylene-Blue/Sodiumtetraborate to give sharp results. } The problem we have is that this staining is rapidly fading if the slides } are mounted with media like Entellan or Eukitt, probably due to the Xylene } content. This might be also a problem of SPURR because with other resins } like LR-White or GMA this doesn't happen. My attemps to use SPURR as a } mounting media, followed by heating at 70 degrees overnight, were not very } sufficient because the resin remained sticky for unknown reasons. } } Any comments, eg. about alternative mounting media, are welcome. } } Jens } } } } --------------------------------------------------------------- } Dr. Jens Buecking Tel. +49-(0)421-218 3745 } University of Bremen Fax. +49-(0)421-218 4620 } Dep. of Biology Email jbueck-at-biologie.uni-bremen.de } Leobener Str. - NW2 } 28359 Bremen } ---------------------------------------------------------------
================= C. John Runions Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
Publication of the Proceedings of the 15th Pfefferkorn Conference, Silver Bay 1996
We are pleased to report that publication of these proceedings is now well advanced. It is expected that the final stages of preparing the material for the printer will be rapid and that printing will begin shortly. We are informed that the Proceedings of the 13th and 14th Pfefferkorn Conferences are ready for printing. Peter Hawkes, Joachim Frank and Owen Saxton, editors of PF15.
I wish to establish the dates and site of the FIRST MEXICAN CONGRESS ON ELECTRON MICROSCOPY. It may well have been held in 1992 but this is not sure. If anyone has firm information, please let me know, any photocopies of abstract booklets etc will be very welcome. Peter Hawkes hawkes-at-cict.fr Fax: (+33) 562 25 79 99
Hi folks - The MSA/Lawrence Hall of Sciencs middle school microscopy manual will be published in a few months. The LHS editor has asked me for microscopy-related quotations to use in publicity and in the manual itself. Do you have any favorites, from the great or the not-so-great? I'm looking for sentences, not paragraphs...
Thanks! Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
{fontfamily} {param} Geneva {/param} {smaller} Dear Electron Microscopists:
This is an announcement and call-for-paper for a book entitled " {bold} Progress In Transmission Electron Microscopy {/bold} ". This book belongs to a "Frontiers of Science and Technology for the 21st Century" book series planned and implemented by The Association of Chinese Scientists and Engineers-USA (ACSE), and will be published by Tsinghua University Press (China). Here I would like to invite your valuable contributions.
The book will be in English and ISBN code will be obtained for international distribution. The purpose of this TEM book is to promote international exchange and communication in TEM field. Being a chapter author of this book, your ideas, scientific findings and achievements, as well as your admirable accomplishments in TEM field will be best known by electron microscopists in the world.
Two volumes are planned for the book: {bold} I. Concepts and Techniques {/bold} ; and {bold} II. Applications in Materials Science {/bold} . The contributed papers/chapters may include review articles, scientific findings and achievements, new concepts, designs, methods, technologies, or summaries of research and development experience. One or more chapters on certain subjects can be contributed by each author (and co-authors). The prospective authors will be from international TEM communities. Please contact me as soon as possible if you would like to have your direct contribution, or you want to recommend chapter authors. Should you have any questions or you want detailed information, please feel free to contact me, or soon you can visit our wibe site http://www.acse.org.
Salzburg, 10th Febr, 1998, local time 03.05 a.m. Yes, I am still working
Dear Ronnie, still your posting came in:
IMO, it (time of "freshness") depends on=20 -the quality of Glutaraldehyde used (original supply bottle with or w/o = dimers or even polymers, check this with the data sheet provided by the = supplier) -the alkalinicity of your solution (the higher pH, -say pH 7.0------7.6 = would be the range of GA-solutions used, the faster you'll get a = polymerization of the aldehyde/s) -how you store the solution (RT, chilled -at- 4 degr.C), tightly = closed/sealed or perhaps if/wether you provided sometime a glass vial = with a snap lid or stopper that doesn't/didn't seal tightly (I have seen = this: after 1 month in the refrigerator, half of the provided solution = was evaporated from the vial due to water condensation in the fridge)
In our lab GA-containing fixatives are prepared freshly (-at-100 ml), which = will last for about 1 week, sometimes 1 and 1/2.=20 If sent out to outdoor facilities I would suggest to either give the = vials (containing approx. 5 ml) an expiration date (say 2-3 weeks if = properly stored), if greater amounts (say 50-100 ml) are provided, = expiration would be 3-4 weeks, if stored properly. BUT: you don't know (do you know??) what "folks out there" are able to = do with your fixatives and vials you provided! (sometimes I got aware of = this fact by "seeing it"). Another question with respect to this is = sampling behaviour in an outdoor facility (e.g. operating theatre) if = you don't have a clue how and when your specimen was handled/put into = the fixative.......
My experience using "some or any " supply(ier) of glutaraldehyde = sometimes was "bad fixation, poor infiltration and polymerization of = resin, poor sectioning performance, and last but not least, bad = ultrastructural morphology (which could be seen before EM-viewing by the = staining properties of semithin sections too). Therefore, check the = source as well as the quality of your GA very carefully....
Just only my 0.2 cents hope this helps very best regards and "Good morning all"
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: Ronnie Houston[SMTP:rhh1-at-airmail.net] Gesendet: Montag, 09. Februar 1998 22:58 An: microscopy-at-sparc5.microscopy.com Betreff: Glutaraldehyde fixn - how fresh?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
How fresh should the working dilution of 2% buffered glutaraldehyde be for diagnostic TEM? Our biopsy material is infrequent, and we don't want to make up quantities that would end up being discarded before viable use. Thanks Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children Dallas
Jennifer Willmott wrote: ============================================== Does anyone know of any sources for having basic SEM analysis done without EDX capabilities? Are the images available in a digital format for distribution? I would also like to know the hourly rates and turn around time if possible. =============================================== There are a number of independent analytical and testing laboratories in the USA ( I assume that is where you are located) offering these kinds of services, both with and without EDS capabilities.
When I want to find something in the EM world, one of the first resources I reach to is that of Microworld Resources at {http://www.mwrn.com/} and in this case, you will find a nice listing of laboratories offering EM related services.
Another excellent resource, also listing services firms is the Microscopy Vendor Data Base at {http://www.kaker.com/} .
You can also find out the names of ACIL member laboratories offering SEM services via the ACIL website at {http://www.acil.org/} . ACIL is the professional, scientific, and trade association for independent testing, analytical, and research laboratories.
And for those laboratories, at least those in the USA, offering such services that are accredited to the standard of ISO Guide 25 by A2LA (American Association for Laboratory Accreditation), if accreditation is important to you, you might want to look at {http://www.a2la.org/} to help you narrow down your search.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
{fontfamily} {param} Geneva {/param} {smaller} Dear Electron Microscopists:
This is an announcement and call-for-paper for a book entitled " {bold} Progress In Transmission Electron Microscopy {/bold} ". This book belongs to a "Frontiers of Science and Technology for the 21st Century" book series planned and implemented by The Association of Chinese Scientists and Engineers-USA (ACSE), and will be published by Tsinghua University Press (China). Here I would like to invite your valuable contributions.
The book will be in English and ISBN code will be obtained for international distribution. The purpose of this TEM book is to promote international exchange and communication in TEM field. Being a chapter author of this book, your ideas, scientific findings and achievements, as well as your admirable accomplishments in TEM field will be best known by electron microscopists in the world.
Two volumes are planned for the book: {bold} I. Concepts and Techniques {/bold} ; and {bold} II. Applications in Materials Science {/bold} . The contributed papers/chapters may include review articles, scientific findings and achievements, new concepts, designs, methods, technologies, or summaries of research and development experience. One or more chapters on certain subjects can be contributed by each author (and co-authors). The prospective authors will be from international TEM communities. Please contact me as soon as possible if you would like to have your direct contribution, or you want to recommend chapter authors. Should you have any questions or you want detailed information, please feel free to contact me, or soon you can visit our wibe site http://www.acse.org.
I'm in the process of buying a new ion mill and would appreciate any opinions on VCR's new XLA/2000, the Technoorg-Linda IV3 and the Bal-tec RES010. I have most of the technical data, but information about maintanace, how user friendly the system is and your general level of satisfaction with your ion mill will be appreciated. Anything you don't like about the system you have?
Please reply to me directly, I will compile a summary for the list server if anybody is intersted.
Many thanx
Sara Prins Surface and Structure Analytical Services Division for Materials Research CSIR PO Box 305 Pretoria South Africa TEL +27 12 8413974 FAX +27 12 8414395
Ziel Rainer (Tel 49(0)6022-812645) wrote: } } Dear all, } } I am looking for an image analysis for a research department. It should be } based on a Windows PC, easy to handle and programmable by a macro language. It } should be easely addapt to different problems. It should have different } algorithem for detection/segmentation as well as the separation of particles. } The image analysis will be used for light microscopy as well as for SEM and } TEM. } } Are there any papers, where image anaysis programs are compared? } } Which is a good listserver for image analysis? } } Are there any suggestions for a good image analysis program? } } Kind regards } } Rainer Ziel } } R.Ziel-at-Akzo.NL
If You have a fast PC with at least 64 MB RAM and Unix running on it You should consider what is probably the most advanced programmable image processing system available for free:
http://www.khoral.com/ ftp://ftp.khoral.com/
Hope that helps
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi folks - The MSA/Lawrence Hall of Sciencs middle school microscopy manual will be published in a few months. The LHS editor has asked me for microscopy-related quotations to use in publicity and in the manual itself. Do you have any favorites, from the great or the not-so-great? I'm looking for sentences, not paragraphs...
Thanks! Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
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Hello all, What is the best way to visualize the HRP stained tissue at EM level? Tissue is run up in EPON and the thick section shows well defined areas at light level. Thanks...
Neelima Shah Electron Microscoppy Core Facility University Of Pennsylvania
Caroline Schooley of MSA recently asked for favorite quotations regarding the microscope and microscopy. I sent her the following quote which I read years ago I think in the frontispiece of a microscopy book: "Where the telescope ends, the microscope begins. Which of the two has the grander view?" I have lost the origin of this quote. I think it might have been Lord Kelvin but I'm not sure. Does anyone out there know the origin of this quote? Thanks.
Roy Christoffersen Texas Center for Superconductivity 3201 Cullen Houston, TX 77204-5932 roy-at-bayou.uh.edu (713) 743-8273 FAX: (713) 743-2787
Doug, I use Thermanox circular coverslips. They fit perfectly into 24 well tissue culture plates. They have a coating on one side for cell adhesion and they're sterile. I have used them for several cell types. Futhermore, they tolerate solvents and dehydrants used for TEM. I have cut them without too much difficulty in situ in plastic and also popped them off in liquid nitrogen. Several vendors handle them. Hank Adams Electron Microscopy Lab New Mexico State University Las Cruces,NM 88003 phone: 505-6463600 fax: 505-6465665
Can anyone on the listserver help with this one? I'd love to use it! CS } } I am happy to oblige you with one of my favorite quotations } regarding the microscope as follows: "Where the telescope ends, the } microscope begins. Which of the two has the grander view?". Unfortunately } years ago I lost the origin of this quote and I would love to determine its } origin. I believe it may have been Lord Kelvin, but I am not sure. If you } could uncover its origin please let me know. Thanks. } } Roy Christoffersen } Texas Center for Superconductivity } 3201 Cullen } Houston, TX 77204-5932 } roy-at-bayou.uh.edu } (713) 743-8273 } FAX: (713) 743-2787
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Hello all, What is the best way to visualize the HRP stained tissue at EM level? Tissue is run up in EPON and the thick section shows well defined areas at light level. Thanks...
Neelima Shah Electron Microscoppy Core Facility University Of Pennsylvania
Ronnie- considering you are working with biopsy material, you should use FRESH fixative. fixatives go bad quite rapidly once in a buffer. concentrated glutaraldehyde (50-70%) is much more stable, and can be stored for probably over a month (check with your supplier) and still work well as a fixative -Mike
On Mon, 9 Feb 1998, Ronnie Houston wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } How fresh should the working dilution of 2% buffered glutaraldehyde be } for diagnostic TEM? } Our biopsy material is infrequent, and we don't want to make up } quantities that would end up being discarded before viable use. } Thanks } Ronnie Houston } Cytochemistry & Molecular Pathology } Texas Scottish Rite Hospital for Children } Dallas }
I recall there were some papers published by Marc Horisberger back in the late 70's about milk fat globules viewed in SEM (he was then working for Nestle - of course!). I believe one of them was in J. Histochem. Cytochem. He also used gold particles to label the globules, BTW. Sorry no specifics. Using this in a citation index might help you get started on the right thread. I hope this helps. Regards, Michel
**************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
} Hi folks - } The MSA/Lawrence Hall of Sciencs middle school microscopy manual } will be published in a few months. The LHS editor has asked me for } microscopy-related quotations to use in publicity and in the manual itself. } Do you have any favorites, from the great or the not-so-great? I'm looking } for sentences, not paragraphs... } } Thanks! Caroline }
Hi Caroline,
A couple of my favourites:
********************************************* Faith is a fine invention For gentlemen to see; But microscopes are prudent In an emergency.
Emily Dickinson (1830-1886) Poems, Second Series ca 1880 XXX ************************************************
In the age of One World, the power of the microscope will be one doesn't know how many times greater that that of [the instrument of] today. [Viewed through the instrument of today] an ant looks like an elephant. [Viewed through the instrument of] the future, the size of a microbe will be like that of the great, skyborne p'eng bird.
K'ang Yu-wei (1858-1927) Ta T'ung Shu: The One-world Philosophy of K'ang Yu-wei transl. L.G. Thompson (1958) London: Allen & Unwin
A microscope is the same as a telescope - you just point a microscope down.
Peter Sewell, LAB-6 Inc., (1940? - ) (1984), on the event of my being hired for a position in Peter's electron microscopy lab. The only marginally relevant experience I had at the time was at an astrophysical observatory.
-------------------------------------------- Michael Cammer email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 http://leper1.ca.aecom.yu.edu/aif/ --------------------------------------------
On Mon, 2 Feb 1998, Goodhouse, Joseph wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } On Fri, 30 Jan 1998 kszaruba-at-MMM.COM wrote: } } } A colleague of mine is looking for a means to fluorescently label } } bacteria prior to seeding onto a surface for testing. The dye must } } not interfere with normal functioning of the bacteria including } } adherence and growth. The result would be viewed under confocal or } } regular fluor. LM. Does such a label exist? }
We're looking for an independant service person to help us maintain our Hitachi H-7000 TEM. Scope is located in Denver, Colorado. please contact Mike Rock {merock-at-du.edu} TIA
Has anyone used an Alps printer to make EM hardcopy? I have been gathering information about the Alps MD-1000 and 2300 models but have some questions; longevity of the prints, image quality (mostly black and white) and reliability? The price seems attractive when compared to a dye sub unit.
Any feedback would be appreciated. Thank you, Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
I received a variety of replies to the post concerning service contracts. = Most of these replies were from laboratories that are located in continenta= l USA. Some replies echoed that they thought that service contracts were = not cost-effective but a *thinly disguised rip-off - especially when one = finds in dealing with a wide range of EM and non EM equipment* that the = non-EM contracts subsidize the cost of EM contracts. For example, = *manufacturers simply calculate the value of the service contract NOT on = the complexity and service requirements of the particular item of = equipment, but simply on a percentage of the value of the equipment. So I = am quoted the same figure for a purely electronic item where 'maintenance' = consists of dusting circuit boards as for an equal value EM where the = maintenance needs are far greater due to the greater mechanical -vacuum = -pneumatic -electronic complexity of the device.* Others valued the = expertise and speed guaranteed by original vendor contracts (*. . . = instrument manufacturers will give priority to their contract customer*). = There was at least one example of long delays (approximately two months) = in obtaining service from the original vendor when no longer under = contract ( * . . . is due to the fact that instrument manufacturers will = give priority to their contract customers*). Even under contract, = original vendors are getting =A1tough=A2 (i.e., charging for expenses) = when the problems stem from other than their equipment (e.g., power = supply). That is understandable.
At least several replies warned that some third parties will bid on = equipment that their staff was not familiar with or were not able to work = on. Either way, third part vendors are unable to carry a satisfactory = inventory of parts and must purchase parts from the original vendor. For = example, *It is critical for prompt service that the engineer can arrive = with replacement boards in hand, swap the boards to revive the scope, and = take the defective boards back home to fix at their leisure. If the 3rd = party agency can't maintain a complete inventory of the needed computer = parts, you are off-line for as long as it takes to fix each board.* This = can cause delays in service that are crippling to a small laboratory.
Service providers did agree *there is a lack of third party service = providers [but] that being said, you should still be able in most cases to = reduce your maintenance costs by 20% by considering third party sources.* = One advantage is that *most small service providers are much more flexible = in their contract services, and can usually provide services tailored to = your maintenance needs - i.e., can adjust their contracts to your = requirements for preventive maintenance, emergency service and replacement = parts needs . . . [And are] always willing to provide customized = contracts designed to reduce your maintenance costs.*
Many replies discussed the role of =A1insurance companies=A2 in providing = service. For example, *A new creature on the market, these organizations = seek to justify themselves by providing a reduction in service costs by = reducing the 'insurance' inherent in a service contract and by trying to = artificially increase competition. They basically work by providing = service on a billable, rather than contract, basis. They will use = whatever service provider they deem reasonable and cheap. When you sign = up with them, you are turning over to them any and all decisions over who = will actually provide service on your instrument and what time frame the = service will be performed in. . . . There simply aren't that many service = providers available to foster the competitive pricing that this approach = requires. When you go with an insurance company, you will not be on [a] = service contract with the manufacturer. * A similar beast is the =A1insurance=A2 that works on billable not = contractual basis. Is this not the same as demand service with a third = party in between? Anyway, this arrangement, is supposed to be transparent = to the user, *in the sense that we just call the usual service provider = when the instrument is down, and the service provider bills XXX directly. = In this way, the quality of service should not be affected. However, I = already experience two shortcomings: One due to the fact that if the = estimated cost of repair is above $5,000 - prior approval from XXX is = required to conduct the service. If the cost is substantially higher, XXX = may decide to get a "second opinion" from a repair service of their choice = (imagine the delays and consequences of having many "experts" try their = hands at your fine instrument!).* There were stories of intolerable = delays, quotes, and more quotations, second opinions and equipment that = remains unrepaired and awaiting more paper shuffling with this type of = service. Not something many clinical or commercial laboratories are able = to tolerate. =20
In summary, original manufacturer service contracts, (not demand service) = although expensive, are favored for computerized electron microscopes, = third-party service may be cost effective for older or less-sophisticated = instruments, and there is general dissatisfaction with insurance-type = service arrangements due to the slowness of repair.
To all that responded, thank you.
Peter O. Steele, Ph.D., PMIAC, Special Anatomic Pathology, All Children's Hospital St. Petersburg, FL, USA
Disclaimer: The opinions expressed are my own and not necessarily that of = my employer.
Why has not Man a microscopic eye? For this plain reason, Man is not a Fly. Say what the use, were finer optics giv'n, T' inspect a mite, not comprehend the heav'n.
Alexander Pope 1733.
"One can be fooled by appearances, which happens only too frequently, whether one uses a microscope or not." Voltaire in Micromegas
"We'll try it," the professor said to me, grimly, ' with every adjustment of the microscope known to man. As God is my witness, I'll arrange this glass so that you see cells through it or I'll give up teaching. In twenty-two years of botany, I -' He cut off abruptly for he was beginning to quiver all over, like Lionel Barrymore, and he genuinely wished to hold onto his temper; his scenes with me had taken a great deal out of him.
So we tried it with every adjustment of the microscope known to man. With only one of them did I see anything but blackness or the familiar lacteal opacity, and that time I saw, to my pleasure and amazement, a variegated constellation of flecks, specks, and dots. These I hastily drew. The instructor, noting my activity, came back from an adjoining desk, a smile on his lips and his eyebrows high in hope. He looked at my cell drawing. "What's that?" he demanded, with a hint of a squeal in his voice. "That's what I saw, " I said. "You didn't, you didn't, you didn't!, he screamed, losing control of his temper instantly, and he bent over and squinted into the microscope. His head snapped up. "That's your eye!" he shouted. "You've fixed the lens so that it reflects! You've drawn your eye!" James Thurber in University Days in My Life and Hard Times.
The texture of Cells of Cork and of some other frothy Bodies could not be so curious, but that possible, if I could use some further diligence, I might find it to be discernable with a Microscope. ... me thinks, it seems very probable, that Nature has in these passages, as well as in those of Animal bodies, very many appropriated Instruments and contrivances, whereby to bring her designs and end to pass, which not improbable, but that some diligent Observer, if help Microscopes, may in time detect.
Robert Hooke in Micrographia: or Some Physiological Descriptions of Minute Bodies made by Magnifying Glasses, 1665.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
I agree for the most part with these comments, most particularly about long-term storage on magnetic media (even Macworld had a warning about that).
The problem of file format standards is perhaps the more important issue, however. Perhaps this is a something that the microscopy community in particular, and the scientific community in general can accomplish some good? Instead of just talking, get together with the instrument manufacturers, and agree on industry standards (like was supposedly done with the jpeg and mpeg standards), and use some marketing muscle to get the software industry to follow them.
Researchers may not be big players in industry decisions, but perhaps we only need to get the ball rolling, and the rest of the mountainside will follow?
I am more sanguine about the life expectency of CD-ROMs. There is a *big* consumer market for audio CDs, and this will keep CD-ROM readers available for a long time (turntables, phono cartridges, and LPs are still available in stores due to analog HiFi nuts). So, archiving images on CD-ROMs isn't a big worry.
Phil
} You have hit the nail directly on the head...I could not have said it } better. As the technology develops, there will be a necessity to } constantly transfer onto up-to-date storage media important data that you } have archived. That is probably one advantage of film...it lasts a long } time, and would be able to be re-printed, or simply scanned and digitally } output, for the forseeable future. But I certainly do not expect that in } the year 2010 it is a guarantee that all my lab's CD-ROMs filled with TEM } and SEM images will be able to be read out on anything but some clunker CD } player we have kept around for that purpose. Of course, by that time I will } be retired, and probably won't give a hoot... ;-). } } Larry } Dr. Lawrence F. Allard } Senior Research Staff Member } High Temperature Materials Laboratory } Oak Ridge National Laboratory } 1 Bethel Valley Road } Bldg. 4515, MS 6064 } PO Box 2008 } Oak Ridge, TN 37831-6064
[...] } } In 20 years, CD-ROM players will be as common as 8-track players } } in new cars. [...] } } Finally, in 20 years, many of the formats you are storing images } } on will no longer be supported. How many of your applications } } can read ILBM and other Amiga format files? /usr/image files? Over } } the next few years, common formats, including .gif and others } } will become either vanishingly rare or astonishingly mutated. } } Remember that neither GIF nore JPEG formats are "standard" for the } } net -- that has gone to PNG. There are early TIFF files around } } that many new applications cannot read because of the } } nonstandardization of compression in "standard" TIFF. } } Those of you who know UNIX freeware know that the famous } } libtiff library supports only a subset of TIFF variants. } } } } billo
Dear List, We have been using Sanyo Cadnica NL5100 flashlights, and they are so reliable that the last person who knew where to order them from has left before we needed to reorder. However, it is now time to replace the few which have died or evaporated, so does anyone on this list know where to order them (or where to order even better ones)? TIA. Yours, Bill Tivol
Hi folks - The MSA/Lawrence Hall of Sciencs middle school microscopy manual will be published in a few months. The LHS editor has asked me for microscopy-related quotations to use in publicity and in the manual itself. Do you have any favorites, from the great or the not-so-great? I'm looking for sentences, not paragraphs...
Here is my favorite.
...by the help of Microscopes, there is nothing so small, as to escape our inquiry; hence there is a new visable World discovered to the understanding ....By this the Earth it self, which lyes so near us, under our feet, shews quite a new thing to us, and in every little particle of its matter, we now behold almost as great a variety of Creatures, as we were able before to reckon up in the Whole Universe it self....
Robert Hooke (Micrographia, 1664)
Bill Monroe EM Center Mississippi State University
} How fresh should the working dilution of 2% buffered glutaraldehyde be } for diagnostic TEM? } Our biopsy material is infrequent, and we don't want to make up } quantities that would end up being discarded before viable use. } Thanks } Ronnie Houston } Cytochemistry & Molecular Pathology } Texas Scottish Rite Hospital for Children } Dallas
Storage life of Glutarldehyde depends upon a number of factors including:
1) type of buffer 2) ph of buffer 3) temperature of storage enviroment
Our suggestion, based on your infrequency of use is to make your solution fresh each time. As little as 50 mL of 2% buffered Glut can be prepared from one 2 mL ampule of 50% Glut. Of course we at Ladd would be happy to supply Glutarldehyde in any concentrations you would like. Please feel free to call me (1-800-451-3406) if you wish to discuss your problem in greater detail.
Dr. Charles Duvic
Ladd Research 13 Dorset Lane Williston, VT 05495 tel 1-800-451-3406 (US) tel 1-802-878-6711 (outside the US) fax 1-802-878-8074
My Initial Question: -------------------------------------
Here is a positive microscopical quotation and a negative one. Note the dates. The quotations and their journal citations are from the preface to Harold F. Schaeffer, Microscopy for Chemists, Van Nostrand, New York, 1953.
[The microscope is] "man's noblest, supreme, and most far-reaching tool."
Adrianus Pijper, South African Journal of Science 26, 58-72 (1939), cited in J. Royal Microscopical Society 62, 36-50 (1942).
"It is rather remarkable how slow American chemists have been in realizing the importance of the microscope as an adjunct to every chemical laboratory. . . . [The microscope is] as much a necessity in every analytical laboratory as is the balance."
When dealing with fading of epoxy sections you must consider at least, at least 3 things: 1) The pH of your mounting medium 2) the chemical activity of your stain 3) the little known, but extremely important, continued chemical reactivity of the epoxy in your sections.
Mounting media come in many differing versions. Try one with a pH near 7.0 such as Cytoseal. The "blues" engage in redox shifts. Basically you cannot cancel this property out. At least, 10% of the epoxy in your sections is not polymerized, and these monomers are free to engage in chemical activity. Therefore, it is never a good idea to use epoxides as a mounting medium as they may encourage color changes or fading in the dye. Study your mounting media at hand. Also try Cytoseal. Add a little bit of the dye to a few mls of your liquid mounting medium in a test tube. Check for fading. Some may taake a week, some just hours, depending on the dye-medium interaction. Make sure, sure, sure, that your blocks are very well polymerized (and of course, infiltrated). Looking at all this may give you an idea as to what is going on. You may even find that really good infiltration with extended polymerization coupled with a mounting medium that is neutral will help to a great degree solve your problem. If not, please contact me again, because I know of some other, but more complicated attacks on the problem. So long, Hildy
"...can the human soul be glimpsed through a microscope? Maybe, but you'd definitely need one of those very good ones with two eyepieces."
Woody Allen
quoted at the beginning of B.A. Palevitz et al. (1981) Protoplasma 109: 23-55
"You can observe a lot by watching."
Lawrence Berra, as quoted in Sports Illustrated, vol. 60 (no. 14), p. 94, 2 April 1984
quoted at the beginning of B.A. Palevitz and P.K. Hepler (1985) Planta 164: 473-479
cheers, Rosemary
Rosemary White Department of Biological Sciences Monash University, Melbourne, Victoria 3168, Australia phone 61-3-9905 5670 fax 61-3-9905 5613 email r.g.white-at-sci.monash.edu.au
I am interested in measuring density gradients within a 100 um granule....more specifically, i need a method of determining the density profile of this granule. Methods that have been proposed include: stereology methods, conductivity measurements, and light intensity measurements through a microtomed section. However, i don't have any details on these methods. Any suggestions or possible references would be greatly appreciated!!!
Thank you in advance!!
Michael Mandanas Particulate Materials Center Pennsylvania State University 218 IMRL Bldg. University Park, PA 16802
We have a problem with embedding whole lenses in Spurrs because the embedding medium is remaining sticky close to the lens, leaving the block virtually impossible to section. Any suggestions would be welcome?
We make gold-conjugated lipids which might be useful for this - they work for labeling liposomes (see Adler-Moore, J. 1994. AmBisome targeting to fungal infections. Bone Marrow Transplantation, 14, S3-S7; you can also see the 1996 MSA abstract about gold-labeled liposomes as a pdf file on the web at
Dear all, We have a periodic table in our TEM lab with X-ray energies and other data given for each element. This is very useful for quick reference and I would like to get hold of one for my office. I have no idea where it came from, however, and don't know if any of the manufacturers of EMs or EDX systems produces such a thing these days. Please contact me if you know where I could get such an item.
Thanks
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Looking for any references / recommendations for stage heaters (i.e. 37C - 50C) for Light Microscope systems. (I realize this is a repeat of previous discussions but I didn't keep the info and now I have a user who wants one.)
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"CONGRESS.SYS Corrupted: Re-boot Washington D.C. (Y/N)?"
I could be doing great things, if I weren't so busy looking at* little things.
I got this from a post card I bought on Fisherman's Warf in San Francisco many years ago. Author Unknown
*original was "so busy doing little things"
} -----Original Message----- } From: Microscopy-request } [SMTP:Microscopy-request-at-sparc5.microscopy.com] } Sent: Monday, February 09, 1998 4:01 PM } To: microscopy } Subject: Quotations? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Hi folks - } The MSA/Lawrence Hall of Sciencs middle school microscopy } manual } will be published in a few months. The LHS editor has asked me for } microscopy-related quotations to use in publicity and in the manual } itself. } Do you have any favorites, from the great or the not-so-great? I'm } looking } for sentences, not paragraphs... } } Thanks! Caroline } } Caroline Schooley } Educational Outreach Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/ } }
Dear all, We have a periodic table in our TEM lab with X-ray energies and other data given for each element. This is very useful for quick reference and I would like to get hold of one for my office. I have no idea where it came from, however, and don't know if any of the manufacturers of EMs or EDX systems produces such a thing these days. Please contact me if you know where I could get such an item.
Thanks
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
A neighboring pathology/histology was CAP inspected yesterday and were informed by CAP inspectors that no pre-written, prelabeled, slides or cassessts are allowed? I had not heard of this. The Pathologist is ever so happy as she does not approve of the prewritten slides or cassetts. She said that histotechs make too many mistakes and she has "caught" them. (not here, but in her previous, another state, institution employed). She said that even with another tech, sometimes two different techs, compareing blocks against sectioned/stained H&E, techs turn out slide with mistakes, mistakes! She prefers sectioning one block, and labeling that slide when sectioning. Gosh, is it just me or are we regressing instead of progressing? Any imput would be appreciated. But what I want to know, is it a CAP ruleing not to use these prelabeling devises anymore? Or a recommendation or what? I know that CAP strongly recommends/suggest that a time clock be placed to date and time when frozen section arrives and date & time when reported. Teresa
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Can anyone provide the source of these nice quotes? You can reply to me at schooley-at-mcn.org.
} Two of my favorite quotations are given below. Since they were picked } up at workshops or conferences many years ago, I have unfortunately } forgotten the attributions. Both "quotations" are paraphrases of the } original statements taken from memory. } } 1) "One who sees more, knows more." (Quotation in reference to the } basic rationale for increasing resolution in microscopes.) } } 2) "Taking a photograph of a specimen using a scanning electron } microscope is like taking a picture of a rose.... using an atomic bomb as } the flashbulb." (Speaker was pointing out that examination of some } specimens in a SEM is not exactly a gentle procedure, hence the need } for good specimen prep. I definitely remember this statement from the } early 1980's by someone conducting the Lehigh summer basic SEM } course.) } } If you find these useful, please let me know. } } Michael M. Craig } Department of Biomedical Sciences } Southwest Missouri State University } Springfield, MO 65804-0094 } mmc519f-at-wpgate.smsu.edu
I am point counting with the electron microprobe on polished thin sections of sandstone, and am using an image analysis program to determine porosity (grey levels) and mineral count (X-ray). I would appreciate hearing from anyone with information or references on this method.
Regards,
Bob MacKay Robert MacKay Department of Earth Sciences Dalhousie University Halifax, Nova Scotia, Canada B3H 3J5 Tel: 902 494-7087 Fax: 902 494-6889 e-mail rmackay-at-ac.dal.ca
Dear All, I am looking for some simple (possibly public domain) software for energy dispersive x-ray analysis which is capable of full standardless quantification of EDX spectra obtained from a TEM. This sounds trivial but I do have a special requirement. I wish to enter the spectrum data through the keyboard of the computer on which the software is installed. i.e. I wish to enter detector take-off angle, accelerating voltage, elements detected, line type and intensity, sample thickness and density etc through the keyboard and then allow the software to perform a fully ZAF corrected standardless quantification. Does anybody know of any software that can do this. Your answers are appreciated.
FEI Company in Hillsboro, Oregon has the following position available:
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FEI Company Hillsboro, Oregon (Portland metro)
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Each employee at FEI works in an open, team-oriented environment and is given the opportunity to fully apply their skills and intellect to a wide variety of challenging problems in manufacturing, engineering or business. Working alongside some of the world's foremost experts in the area of field emission technology, both new and experienced employees will lean and gain by the opportunities of working at FEI Company.
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Please submit resume to Lisa Olivia either by FAX: 503.640.7509 or email: lolivia-at-feico.com.
Lisa Olivia Recruiter FEI Company PH) 503.844.2601 FAX) 503.640.7509 email: lolivia-at-feico.com
I don't think that is a CAP deficency, however an inspector can make a comment or recommendation. I've done several Anatomic CAP inspections myself, I think the issue of pre-labeling slides comes up more so than with cassettes in the gross room. I think if you get in several specimens at a time and are preparing them with cassettes for the pathologist, it would be difficult to do one at a time. In fact I don't know many pathologists who would be patient enough for waiting for you to make up cassettes one case at a time. The person doing the gross, be it a pathologist or PA, should also want to check the number on the cassettes against the number on the requisition to ensure accuracy. Like anything else, if your medical director insists you should do this practice, well then you don't have a choice.
There are a number of fundamental bits of infornation involved in calculating the correction factors involved in microbeam analyses (e.g. ionization cross sections, electron transition probabilities, mass absorption coefficients, etc., etc.). In general, these factors are much more accurately known for the K-series lines than for L and M lines, and so analyses with K lines are usually likely to be more accurate, whether standards or standardless methods are employed.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Can Anyone recommend a Canadian supplier for nylon lint free gloves. The type I am interested in are the tailored fit style (they look like butlers gloves) not the baggy type. I used to get them through JBEM services but can no longer.
From my SEM, I'm sending images to people, .tiff images inserted into Microsoft's Power Point. A problem is that a single .tiff image is over 1 megabyte and inserting 20 or 30 into a Power Point presentation makes for a 20 or 30 megabyte e-mail message and I get all sorts of grief from the people that I send it to. Now, I can convert these images into .jpeg files and the amount of memory that they take is substantially reduced, 1027 kb becomes 126 kb and there is a loss in quality but, depending upon the sample, I can accept the loss in absolute image quality at times. I can view the .jpegs in Thumbs and LViewPro, but the big problem is that Power Point, and I believe Word also, does not let you insert a .jpeg image into a document. So, if I send a .jpeg image to a person I can tell them to get thumbs to view it, by itself, but I'd like to put the .jpegs into a report much like Power Point, where the spaces for graphics and text are ready to go, graphics with explanation. Is there software like Power Point that I can set up a .jpeg presentation? Or, is there another image file that I can convert the .tiff to, instead of a .jpeg, compressing it to a smaller size than a .tiff? Is there someone out there going through this also?
Applied Optical Microscopy (An American Chemical Society Short Course, held in conjunction with PITTCON) Three days and two evenings of total immersion in microscopy; the last day is dedicated to polarized light. A full lab course, with exercises on alignment, contrast techniques, basic measurement, polarized light, and videomicroscopy. Great for biologists, too. An ideal opportunity to discuss your favorite problem with an expert microscopist.
Comments from some of last year's students: "It's so nice to have someone discuss the "why"!" "The course opened=85my eyes to many interesting techniques which I can't=
wait to get back and try!" "An excellent lab class!" "It doesn't matter what your current level of microscopy knowledge is, you will benefit from this course =85. [and] I'll come with you --- there's always more to learn!"
Registration information: February 27, 28, March 1 - ITT Sheraton, New Orleans, LA Course #AOPT9802: $895 for ACS members; $995 for non-members =
For further details: 1. Contact Barbara Foster, course coordinator, at Microscopy/Microscopy Education Ph: (413)746-6931 Fx: (413)746-9311 email: mme-at-map.com 2. See the ACS web site: www.ACS.org/education click Professional Development (side bar) then: short courses then: analytical
Thanks to the many people who responded to my question! For those who might be intersted, the original question is given below followed by a summary of responses.
Karen
} A colleague of mine is looking for a means to fluorescently label } bacteria prior to seeding onto a surface for testing. The dye must } not interfere with normal functioning of the bacteria including } adherence and growth. The result would be viewed under confocal or } regular fluor. LM. Does such a label exist? } } Acridine Orange was tried a while ago for a slightly different } experiment and found to show too much bleeding under confocal. } Also, UV excitation is out. On the Confocal Listserve there was a } discussion some time ago mentioning dyes from Molecular Probes such as } RH414, the Syto dyes esp. #10, Live/Dead, TMRE, as well as others. I } have no personal experience with any of these. Does one stand out for } this purpose? } } Thanks as always, } Karen } } -- } Karen Zaruba, kszaruba-at-mmm.com } Life Sciences Sector Laboratory, } 3M Center Bldg. 270-1S-01 } 3M Company, St. Paul, MN 55144
==============================
Answers Received from Microscopy Listserve and Confocal Listserve:
1. The most popular suggestion by far was GFP! One respondent had experience with an E.coli strain that had been transfected with GFP. After many cell divisions other dyes such as Live/Dead would not carry over but GFP would.
2. Fluorescein diacetate: "The diester is not fluorescent and, being nonionic, can get into the cell. Once in, intracellular enzymes hydrolyze off the acetates, giving fluorescein which is ionic and does not get out very easily. There was a publication in Proceedings of the (USA) National Academy of Science around 1967-8. I tried it just enough to know that it does work. Since then other fluorogenic substrates have been developed, and some of them might work better."
3. Proteins labeled with fluorescein could be fed to the bacteria.
4. Nile Red
5. PkH2 from Sigma. Noted the dye is high intensity, and mammalian cells could be kept alive for several weeks after labeling. There was some loss in intensity with proliferation. "The dye is fixable as long as you do not use detergents or any extracting reagents, such as acetone and alcohols. It intercalates into lipid bilayers and fluoresces strongly with 488 excitation."
6. Syto 16 from Molecular Probes Found this most promising out of Syto's 11-16, nontoxic until exposed to laser when cells often became "PI positive (leaky cellular membrane)."
7. Reference to Dr. Graham Darling's work (for nonspecific fluorochrome): "Canadian Journal of Microbiology (1996): A novel fluorochrome for the microscopic observation of microbial morphology in wet mounts. They (Chan and Darling) synthesized the following compound trans-4-p(p-N,N-dimethylaminostyryl)-N-butoxycarbonylmethylpyridinium and used it to follow bacterial and spore growth for up to seven days. I know that it is commercially available now."
8. Finally a caution that each cell type may be more or less susceptible to the dye toxicity, and one should grow out the strain in the dye, looking for normal morphology, before using the dye. Comments on specific dyes: "The RH dyes and TMRE are membrane stains and tend to make the cells look like little halos, and they work fairly universally (stain all cells - but dead ones don't stain well with TMRE). The nucleic acid stains (green and red Sytos) are nice but these don't all work with all cell types (strange how cells still grow with the stain stuck all over the DNA....)."
Our approach, which our customers believe is a lot more user friendly, is to send 2 or 3 images (usually jpeg) in an e-mail and then overnight mail the 20 or 30 prints and/or files on CD-r or whatever for next day delivery.
I can understand why you would be getting grief sending out a 30 MB mail message. I think the powers-that-be have limited our mail messages to 10 MB which is still a heck of a message. My personal opinion is that mail is probably not the right way to disseminate images unless you just have no other way. I would suggest an FTP server of some kind for distributing the images. If you set up an anonymous FTP service, outsiders should be able to retrieve their own images. I have not tried it, but I think they should even be able to access the files via the Web by pointing to your directory. You should not have to be running a Web server. You can take a look at our site for an example.
There are a number of options for setting up such a service. Personal Web Server for Windows 95 also allows for an FTP server. There are other programs like QVT-Net and War-FTP which will serve up FTP as well. But back to your immediate problem...
You should be able to import JPEG images into Word and PowerPoint - I can in my PowerPoint Version 7. You might have to explicitly load a JPEG graphics filter first. I don't think the older Office products loaded many graphics filters by default. You had to customize the setup to get them. The results should be a file with a name similar to JPEGIM*.FLT in GRPHFLT in the shared Microsoft Apps area. That was the MSAPPS subdirectory under Windows 3.x and is the "Common Files/Microsoft Shared" directory under my 95 version.
Microsoft distributed a product called MS Imager on its Office CD up through version 6 (and mayber later). It now distributes Wang Image with either 95 or Office (or both). They both can process JPEG images. You should be able find those around somewhere or another, or else contact me for further details.
Some TIFF writers support compression. However, it is often not very effective on EM images. Same goes with GIF and PCX.
JPEG gets such good results because it is a lossy image compression algorthm, but it is quite good for most images, especially if you opt for a more faithful rendition. Because some very fine detail is sacrificed they are able to achieve very high compression ratios. I am not aware of any other standard formats that provide near as much compression. You probably will want to get yourself setup to work with JPEG.
BTW, could this be what they meant when they said "the devil is in the details"?
At 02:57 PM 2/11/98 EST, you wrote: } From my SEM, I'm sending images to people, .tiff images inserted into } Microsoft's Power Point. A problem is that a single .tiff image is over } 1 megabyte and inserting 20 or 30 into a Power Point presentation makes for a } 20 or 30 megabyte e-mail message and I get all sorts of grief from the people } that I send it to. } Now, I can convert these images into .jpeg files and the amount of } memory that they take is substantially reduced, 1027 kb becomes 126 kb and } there is a loss in quality but, depending upon the sample, I can accept } the loss in absolute image quality at times. } I can view the .jpegs in Thumbs and LViewPro, but the big problem is } that Power Point, and I believe Word also, does not let you insert a .jpeg } image into a document. So, if I send a .jpeg image to a person I can tell } them to get thumbs to view it, by itself, but I'd like to put the .jpegs into } a report much like Power Point, where the spaces for graphics and text are } ready to go, graphics with explanation. } Is there software like Power Point that I can set up a .jpeg } presentation? Or, is there another image file that I can convert the .tiff } to, instead of a .jpeg, compressing it to a smaller size than a .tiff? } Is there someone out there going through this also? } ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
I am an amateur entomologist/microscopist/naturalist . I am looking for a trinocular microscope with excellent resolution and definition for microphotography purposes with koeler illumination ,magnification is not critical , I would be satisfyed up to 600X or 800X. Clarity and definition being first and formost I am looking for a real bargain , like a university or instituition that is getting rid of equipement at exceptionally low prices . THANK YOU ,RICHARD CLARKE : rclarke-at-total.net.
Kalvin Electron Microscope Lab Rm 1257 W Lenox Hill Hospital 100 E 77th St NY NY 10021
Friends:
YIKES! Anyone who trusts ZIP, Jaz, tape drives, and CD's for archived data is risking disaster!
The only way to go is magneto-optical. PERIOD! ^^^^^^^^^^^^^^^^^
later
-gene
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
Could not help but pass these paragraphs along to you, Caroline. The author is "anonymous".
The Toad
In days of old, those far off times of high romaance and magic, A toad was an enchanted prince, A transformation tragic.
Today the toad is studied as A scientific topic No prince is found, although we look With vision microscopic.
And yet, the prince is there - he's there As clearly as can be. Forget your microscope, my friend, And use your eyes to see!
Nancy R. Smith Director of Operations Microscope And Graphic Imaging Center California State University, Hayward Hayward, CA 94542 http://www.csuhayward.edu/SCI/sem
The Center for Solid State Science at Arizona State University is looking for a unique Academic Associate to manage its new Materials Visualization & Analysis Facility and assist in coordination of education/outreach activities. The Facility will be networked to the advanced imaging and analysis capabilities of the Goldwater Materials Science Laboratories via a Windows NT based operating system providing on-site and off-site network access for education, research and research training. The appplicant must have a B.S./M.S. in Chemistry, Physics, Engineering or a related discipline. At least two-years experience with NT system management, C++, Java, and Visual Basic is strongly desirable. Materials Science, molecular modeling, education/outreach, and higher end computer (e.g., UNIX, SGI) experience are also desirable. The application deadline is April 1st, 1998 or the first of each month thereafter until the position is filled. Send a resume and names of three references to: Dr. Michael J. McKelvy, Chair, Search Committee, Center for Solid State Science, Arizona State University, Tempe, AZ 85287-1704. Arizona State University is an AA/EEO employer.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
New versions of Power Point and Word in Office 97 (and now Office 98) = accept .jpeg images as easily as any other format. I just did this for = a presentation myself.
Good luck,
Tom Thomas C. Isabell, Ph.D. Research Scientist E.A. Fischione Instruments, Inc. tci-at-fischione.com webpage: www.fischione.com
-----Original Message-----
From my SEM, I'm sending images to people, .tiff images inserted = into=20 Microsoft's Power Point. A problem is that a single .tiff image is over =
1 megabyte and inserting 20 or 30 into a Power Point presentation makes = for a=20 20 or 30 megabyte e-mail message and I get all sorts of grief from the = people=20 that I send it to. =20 Now, I can convert these images into .jpeg files and the amount of=20 memory that they take is substantially reduced, 1027 kb becomes 126 kb = and=20 there is a loss in quality but, depending upon the sample, I can accept=20 the loss in absolute image quality at times. I can view the .jpegs in Thumbs and LViewPro, but the big problem = is=20 that Power Point, and I believe Word also, does not let you insert a = .jpeg=20 image into a document. So, if I send a .jpeg image to a person I can = tell them to get thumbs to view it, by itself, but I'd like to put the = .jpegs into=20 a report much like Power Point, where the spaces for graphics and text = are=20 ready to go, graphics with explanation. Is there software like Power Point that I can set up a .jpeg=20 presentation? Or, is there another image file that I can convert the = .tiff=20 to, instead of a .jpeg, compressing it to a smaller size than a .tiff? Is there someone out there going through this also?=20
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.2106.6"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I've been asked to examine the grain = boundaries=20 of calcium metal with an SEM. Can anyone suggest a sample prep=20 technique?? {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Nan H. Laudenslager {/FONT} {/DIV} {DIV} {FONT size=3D2} Specialty Minerals. Inc. {/FONT} {/DIV} {/BODY} {/HTML}
In response to: =============================================== Can Anyone recommend a Canadian supplier for nylon lint free gloves. The type I am interested in are the tailored fit style (they look like butlers gloves) not the baggy type. I used to get them through JBEM services but can no longer. ================================================ These gloves are available from most of the major suppliers of EM consumable products, in addition to SPI, also Pella, EMS, Fullam, and Ladd. You can find them in our electronic catalog on our website given below.
While on the topic of gloves, one question we are asked frerquently is what are the relative advantages/disadvantage of cotton vs. nylon for handling the filament cap, housing, etc. Some seem to believe passionately in one and are anti the other. Even service engineers from the same microscope manufacturer give opposite recommendations!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
A broad summation, but basically true. Here are some different angles:
Manufacturer service: Generally considers providing service a necessary evil. Service personnel are not given an upward migration path, thus leading to high turnover and a generally low rate of service engineer competency. Part charges are generally 1000 - 1500 percent of the market value (99% of instrument problems require the replacement of electronic industry standard parts).
In a system as complex as an EM, manufacturer service engineers seldom carry with them replacement modules. While they can order such parts for rapid delivery, anyone can order those parts for similar delivery - including you or a third party service source.
Third party sources: You take your chances. There are competent third party sources out there, but there are also the incompetents. Experience with the particular instrument you are using is not necessarily a large hurdle, but it could be if the individuals don't have a broad experience. There is, more than likely, nothing special about the design of your particular instrument. The field of EM has not made any revolutionary changes for a long time. Make sure that you have an 'out', a reasonable cancellation clause for contract services.
I am, admittedly, biased towards third party providers, being one. At the same time, I can honestly state that I have saved my customers millions of dollars over the 16+ years I have been in business, and managed to provide a level of service at least equal to the manufacturers.
Third party service providers have a great deal more flexibility in catering to the customer needs than a manufacturer's service organization. Aside from the contractual flexibility, third party service providers bring a competetive ability in labor and replacement part pricing.
'HMO' style providers: These services seek to pit third party providers against each other in order to reduce pricing through competition. All service will be provided through billable service.
This is not necessarily bad. Many of the instruments out there do not require the low latency response time that service contracts provide. These organizations essentially take your active involvement out of the provision of service. Consider the time and expense of your involvement in seeking and maintaining ongoing service for your instruments, you may find that your involvement in the process is more costly than you realize.
However, if you require a high instrument up-time, you must plan on a contractual relationship that can offer you some guarantees.
Instrument service contracts essentially are comprised of a few, differing, components. Expected labor time and expenses, part replacement costs and catastrophic expenses. This latter category is the 'insurance' that bloats service contracts. However, it also amortizes those infrequent, but possibly expensive, problems that contribute to the long term maintenance of an instrument.
If you are fortunate enough to be able to justify the replacement of large capital equipment in a short period of time, you may be best served by the 'HMO' services out there, or perhaps the third party service sources. If that expensive system that you bought will have to last a long time, the manufacturer or a third party service provider will probably serve you best.
The common, middle of the road, answer may well be a carefully chosen third party service source that has a proven track record and a broad experience. That, however, can be hard to find. You will have to take an active role in the provision of service if you choose this route.
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } -------------------------------------------------------------------- } ---. } } I received a variety of replies to the post concerning service } contracts. Most of these replies were from laboratories that are } located in continental USA. Some replies echoed that they thought } that service contracts were not cost-effective but a *thinly } disguised rip-off - especially when one finds in dealing with a wide } range of EM and non EM equipment* that the non-EM contracts } subsidize the cost of EM contracts. For example, *manufacturers } simply calculate the value of the service contract NOT on the At } least several replies warned that some third parties will bid on } equipment that their staff was not familiar with or were not able to } work on. Either way, third part vendors are unable to carry a } satisfactory inventory of parts and must purchase parts from the } original vendor. For example, *It is critical for prompt service } that the engineer can arrive with replacement boards in hand, swap } the boards to revive the scope, and take the defective boards back } home to fix at their leisure. If the 3rd partyService providers did } agree *there is a lack of third party service providers [but] that } being said, you should still be able in most cases to reduce your } maintenance costs by 20% by considering third party sources.* One } advantage is that *most small service providers are much more } flexible in their contract services, and can usually provide } services tailored to your maintenance needs - i.e., can adjust their } contracts to your requirements for preventive maintenance, emergency } service and replacement partMany replies discussed the role of } insurance companies in providing service. For example, *A new } creature on the market, these organizations seek to justify } themselves by providing a reduction in service costs by reducing the } 'insurance' inherent in a service contract and by trying to } artificially increase competition. They basically work by providing } service on a billable, rather than contract, basis. They will use } whatever service provider they deem reasonable and cheap. When you } sign up with them, In summary, original manufacturer service } contracts, (not demand service) although expensive, are favored for } computerized electron microscopes, third-party service may be cost } effective for older or less-sophisticated instruments, and there is } general dissatisfaction with insurance-type service arrangements due } to the slowness of repair. } } To all that responded, thank you. } } Peter O. Steele, Ph.D., PMIAC, } Special Anatomic Pathology, } All Children's Hospital } St. Petersburg, FL, USA } } } } } Disclaimer: The opinions expressed are my own and not necessarily } that of my employer. } } } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
We are using a PC machine for the following discussion.
What version of Powerpoint are you using? We have a similar problem here at PPG. We also have a 1Mbyte limit in our firewall for our Email, -Bummer.
Our standard Office version is two versions old (V4.0). We have one machine that we have Office97 loaded on. If we save a Word document that has a few Tif images in both Word97 and Word version 6 from Word97, then the Word97 document can sometimes be 10-15% of the file size that the Word 6 version takes up. I think that the Powerpoint97 does the same thing. I'm pretty sure what they are doing is saving the files in a compressed mode.
You have two options: 1) update to Office 97 (Word97 Powerpoint97, etc.) or 2) use a compression utility such as PKZip to send the document and have your colleague at the other end decompress it. I've recently zipped a 11Mb Word 6 document that had several images and graphs to about 3Mb doing this.
As to your other point, you can insert a jpeg file into a document if you have the graphics filter for it. The following lines are from my Win.ini file for the jpeg filter:
You can go to the Microsoft site and get this and other graphic and text filters.
However, this does not buy you anything. If you insert a tif or a jpeg image into Word or Powerpoint, then the filter will also uncompress the image to all its glory when it translates it into the document. Sorry.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
From my SEM, I'm sending images to people, .tiff images inserted into Microsoft's Power Point. A problem is that a single .tiff image is over 1 megabyte and inserting 20 or 30 into a Power Point presentation makes for a 20 or 30 megabyte e-mail message and I get all sorts of grief from the people that I send it to. Now, I can convert these images into .jpeg files and the amount of memory that they take is substantially reduced, 1027 kb becomes 126 kb and there is a loss in quality but, depending upon the sample, I can accept the loss in absolute image quality at times. I can view the .jpegs in Thumbs and LViewPro, but the big problem is that Power Point, and I believe Word also, does not let you insert a .jpeg image into a document. So, if I send a .jpeg image to a person I can tell them to get thumbs to view it, by itself, but I'd like to put the .jpegs into a report much like Power Point, where the spaces for graphics and text are ready to go, graphics with explanation. Is there software like Power Point that I can set up a .jpeg presentation? Or, is there another image file that I can convert the .tiff to, instead of a .jpeg, compressing it to a smaller size than a .tiff? Is there someone out there going through this also?
Has anybody had any experience in cutting cryo sections of fixed, decalcified bone (mouse femurs) using a cryostat, for light microscopy? If so I would appreciate some helpful hints. Thank-you, Susie Nilsson C/o Sarah Ellis Research Division Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne, Victoria 3000 Australia
I would like to know how much someone must pay for the 'GATAN DigitalMicrograph' software version 3.x. for education or business. Any information is highly appreciated.
The FTP server has worked very well for us these past few years. I post a client's images (usually as .tiffs) to our firewall-protected anonymous-access site, usually the day after they were produced, and give them a little handout with accessing instructions clearly described. the client usually has a week or two to get them from the site before our system administrator routinely removes them. And yes, once or twice I've had to re-post them because the client was a little late getting them off. As for image archiving, since there's been a fair bit of traffic about this lately, I use CD's. We have a NORAN Voyager system, which produces images as ".greys", a format only Voyager understands, but can also produce copies as .tiffs. My archive CDs (one for each client) contain each image as both a .tiff and the original .grey. The client gets a similar CD. He/she probably can't do much with the .grey, but it serves as as an off-site archive, should something happen to ours. As far as longevity of CDs goes, I think I can reasonably expect them to last at least a decade or two in optimum storage conditions. By that time surely the scientific project the images were acquired for has run its course, to publication or whatever. Besides, I'll be retired or otherwise gone and probably won't care a whole hell of a lot.
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
I am looking for a dry detector for EDAX attached to Philips TEM CM-10. Could you please let me know where I can find one. E-mail addresseas or telephone numbers of the dealers or the manufacturerers is what I am looking for.
} YIKES! Anyone who trusts ZIP, Jaz, tape drives, and CD's for archived } data is risking disaster! } } The only way to go is magneto-optical. PERIOD! } ^^^^^^^^^^^^^^^^^
On the other hand, however, we have had significant problems with a MO drives and disks with disks becoming several cases of disks becoming unreadable after a while and one MO drive which totally died.
Whilst, they are not archiving media, Zips are handy, cheap and, so far, I have had more problems with 1.4Mb floppies than with Zips. The only Zip disk that I have had that died was replaced free by Iomega.
Seems like no media is perfect and multiple copies of data are always a good idea.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
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Why are you sending images in PowerPoint? That is a presentation software package for making slides, handouts, etc. Why not use PhotoShop for archiving and manipulating images. It allows you to do so much more with the images, is cross-plateform, permits saving in all sorts of file formates including JPEG and compressed TIFF. If you don't already have Adobe PhotoShop, I strongly recommend you getting it. Although it is an enormously powerful image processing program, the average microscopist only needs a relatively few features so the learning curve is not too bad. The ability and control you have to adjust gamma, contrast and brightness, image size, cripping, layout using levels, etc. is invaluable in addition to having all kinds of filtering available if desired. Color correction and balance is also at your fingertips if needed. You then are free to export the image to any object oriented program such as a drawing program or PowerPoint for making your final presentation figure.
Debby Sherman, manager Microscopy Center in Agriculture Purdue University West lafayette, IN 47907-1057 765-494-6666 E-mail: sherman-at-aux.btny.purdue.edu
--------------------------------------
From my SEM, I'm sending images to people, .tiff images inserted into Microsoft's Power Point. A problem is that a single .tiff image is over 1 megabyte and inserting 20 or 30 into a Power Point presentation makes for a 20 or 30 megabyte e-mail message and I get all sorts of grief from the people that I send it to. Now, I can convert these images into .jpeg files and the amount of memory that they take is substantially reduced, 1027 kb becomes 126 kb and there is a loss in quality but, depending upon the sample, I can accept the loss in absolute image quality at times. I can view the .jpegs in Thumbs and LViewPro, but the big problem is that Power Point, and I believe Word also, does not let you insert a .jpeg image into a document. So, if I send a .jpeg image to a person I can tell them to get thumbs to view it, by itself, but I'd like to put the .jpegs into
a report much like Power Point, where the spaces for graphics and text are ready to go, graphics with explanation. Is there software like Power Point that I can set up a .jpeg presentation? Or, is there another image file that I can convert the .tiff to, instead of a .jpeg, compressing it to a smaller size than a .tiff? Is there someone out there going through this also?
by gater3.sematech.org (8.7.6/F-1.12) with ESMTP id IAA10792; Thu, 12 Feb 1998 08:27:20 -0600 Received: from franklin.sematech.org by SEMATECH.Org (PMDF V5.1-10 #26085) with ESMTP id {01ITHI4BLJA08WYU2S-at-SEMATECH.Org} for microscopy-at-Sparc5.Microscopy.Com; Thu, 12 Feb 1998 08:27:17 CST Received: from localhost (root-at-localhost) by franklin.sematech.org with SMTP (8.7.6/8.7.3) id IAA05270 for {microscopy-at-Sparc5.Microscopy.Com} ; Thu, 12 Feb 1998 08:27:05 -0600 (CST)
Mark (and everyone else),
If you really feel that jpeg compression is the way to go, you can have your customers view them with a program that they likely already have... called Netscape (or Microsoft Internet Explorer for that matter). You can set up an html file that calls the images up, or let people call them up the images individually from whatever folder they might rest in.
I've been thinking about these things because I am also trying out various ways of sending digital images, at the moment I am still using large tiff format files, and I am putting these on a hard drive that is accessible to everyone on our in-house network. there are still "issues" with this solution, I am still working out file protection issues, but this hasn't been a problem (just a concern).
One reason I haven't gone to jpeg images is that, as you had mentioned, they don't go easily into Microsoft powerpoint... which is where most of my customers want to put them.
Brendan Foran SEMATECH Austin, TX
Standard disclaimer: these are my opinions, and should not be assumed to reflect those of my employer.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From my SEM, I'm sending images to people, .tiff images inserted into Microsoft's Power Point. A problem is that a single .tiff image is over 1 megabyte and inserting 20 or 30 into a Power Point presentation makes for a 20 or 30 megabyte e-mail message and I get all sorts of grief from the people that I send it to. Now, I can convert these images into .jpeg files and the amount of memory that they take is substantially reduced, 1027 kb becomes 126 kb and there is a loss in quality but, depending upon the sample, I can accept the loss in absolute image quality at times. I can view the .jpegs in Thumbs and LViewPro, but the big problem is that Power Point, and I believe Word also, does not let you insert a .jpeg image into a document. So, if I send a .jpeg image to a person I can tell them to get thumbs to view it, by itself, but I'd like to put the .jpegs into a report much like Power Point, where the spaces for graphics and text are ready to go, graphics with explanation. Is there software like Power Point that I can set up a .jpeg presentation? Or, is there another image file that I can convert the .tiff to, instead of a .jpeg, compressing it to a smaller size than a .tiff? Is there someone out there going through this also?
} } YIKES! Anyone who trusts ZIP, Jaz, tape drives, and CD's for archived } data is risking disaster! } } The only way to go is magneto-optical. PERIOD!
I am curious why magneto-optical is preferred over CD (write once)? What size MO?
In our situation, we use all of the above (including MO's of 128 and 230 MB) but prefer CD's for archiving since they may be read by all platforms (using ISO9660).
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Is anyone else interested in lobbying to have Caroline's microscopy quotes list posted to the group? If it isn't too much trouble, of course - but some that were sent to the list have been wonderful (Woody Allen's, especially)...Would anyone be offended at the "wasted' bandwidth?
Just a warning about Microsoft Powerpoint 97. We've had a problem importing tiff files with this software. Hopefully the newer version has corrected the problems
Yeah, but our 2-year-old HP M-O drive will no longer reliably accept its cartridges. These were the 1300 MB cartridges with 650 MB on a side. I hoped that we could find another in the area to back us up, but we seem to have the only one in town. You wouldn't happen to have an HP 1300T drive would you? {G}
I guess we all have to beware.
At 05:12 PM 2/11/98 -0600, you wrote: } } Kalvin Electron Microscope Lab } Rm 1257 W } Lenox Hill Hospital } 100 E 77th St } NY NY 10021 } } Friends: } } YIKES! Anyone who trusts ZIP, Jaz, tape drives, and CD's for archived } data is risking disaster! } } The only way to go is magneto-optical. PERIOD! } } later } } -gene ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
Here is a brief summary of the replies I received about the M&M1999 physical science tutorials:
1. "How about GIF and perhaps PEELS?"
2. Defect recognition and analysis in crystalline materials.
3. Accessing and using on-line crystallogrphic databases
4. SEM Techniques
5. Optimizing the microscope for XEDS and WDS detection
6. XEDS imaging and interpretation, the right ways and the wrong ways.
7. Automation and Remote Control
8. More sample preparation (cross section of ALL methods)
9. Cross section samples with the FIB
10. Spectrum imaging
It looks like some of these clearly overlap and so I may have the basis of a couple of tutorials. If there are further suggestions and/or comments please feel free to send them. If you merely want to vote (for or against) any of the suggestions then that is OK too.
Regards
Jfm
________________________ Note new Area Code (734) ________________________ John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Somewhere I am sure this has been mentioned before, but considering this thread, I mention again....
JPG files are great for reducing the (file) size in order to send them out for "review". This method of image compression is, however, "lossy". That is, image information is lost forever during compression. Various degrees of compression are available for JPGs. At minimum loss it is very difficult to tell something has happened to the image using the naked eye. At maximum compression, the loss is significant and quite visible in most images. It is also my understanding that recalling and saving a JPG repeatedly will, to some degree add more data loss.
For archiving scientific images, especially when the image may later be subject to "image analysis", editing, or close realtime scrutiny, a lossless mode of saving is indicated. TIFF and PCX file types are such examples.
Woody White McDermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com
} } The only way to go is magneto-optical. PERIOD! } } I am curious why magneto-optical is preferred over CD (write once)? } What size MO? } } In our situation, we use all of the above (including MO's of 128 and 230 } MB) but prefer CD's for archiving since they may be read by all platforms } (using ISO9660).
I would NOT rely on MO for archiving because of the lack of a standard disk format. ( Well -- there is an ECMA "standard", but I don't know of any vendors implementing it! )
Not only is it not cross platform, but even changes in the DOS/Windows disk format will break it -- this happened to us going from DOS 3.x to 4 & 5 . MO Disks formatted under 3.x were unreadable under v4 or v5.
Disks are also not necessarily portable if you change disk controllers.
From my experience, neither the MO disk vendors, SCSI controller vendors, or Microsoft seem very concerned with questions of data archive integrity. Their solution to incompatible changes is always the same: "Just backup and reformat"
CDs at least have a standard that you can expect will be supported for many years.
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |--- All power corrupts and obsolete power corrupts obsoletely." - Ted Nelson
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I just received bulk mail from mailto:art_and_mikey-at-justamateurs.com. Has anyone else received the sexsite advertisement??? Even if I was single out somehow, the Microscopy list was responsible somehow because it had the Microscopy text header ...
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I just received bulk mail from } mailto:art_and_mikey-at-justamateurs.com. Has anyone else received the } sexsite advertisement??? Even if I was single out somehow, the } Microscopy list was responsible somehow because it had the Microscopy } text header ... } } } cheerios, shAf } -- } {\/} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {\/} } } Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ } Geological Science's Electron Probe Facility at the University of Oregon } } mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu } } }
We get these from time to time. Luckily most of us hit the "delete" = button from the header details.
Just a reminder NOT to hit reply and type "remove" as it's not you who's = on their list, it's the list server. You'll only succeed in = broadcasting your message to the whole list.
Cheers, Roger
Roger Wallis General Manager Optiscan P/L Confocal Microscopy PO Box 1066 Mt. Waverley MDC Victoria 3149 Australia Tel: (61) 3-9562-7741 Fax: (61) 3-9562-7742 e-mail: rogerw-at-optiscan.com.au URL: http://www.optiscan.com.au ______________________________
} I just received bulk mail from } mailto:art_and_mikey-at-justamateurs.com. Has anyone else received the } sexsite advertisement??? Even if I was single out somehow, the } Microscopy list was responsible somehow because it had the Microscopy } text header ...
We all received it. The list-manager should modify the software so that only identified registered subscribers can submit messages to the server. Other lists have done so.
No disrespect intended, but I can't let this go by unaddressed. My apologies in advance, Nestor.
You state: } "Manufacturer service: } Generally considers providing service a necessary evil. Service } personnel are not given an upward migration path,..."
It appears that to you there is no "upward migration path", which I assume is to mean some nebulous region of importance in one's mind, and that results in a better paycheck. Due to the complexity of EMs, service engineers cannot help but constantly increase their skills and expertise. Our "upward mobility" is always in motion. It doesn't stagnate and the permutation of service problems are infinite, challenging, and enlightening...and it, too, results in better paychecks.
} "...thus leading to high turnover and a generally low rate of } service engineer competency."
Of the 70+ service engineers in the company I work for, the average length of time the company is 10 to 15 years with several 25+ years service, the least 3 1/2 years. Turnover is very rare and to my knowledge has never been due to the lack of upward mobility. I will not take issue with your statment that there is a 'generally low rate of service engineer competency'. I assume you hire the best...so do we. AND manufacturer service engineers are constantly being trained and updated on any EM service modifications and new products.
} "Part charges are generally 1000 - 1500 percent of the market value } (99% of instrument problems require the replacement of electronic } industry standard parts)."
Though the manufacturer list price to non-contract customers is significantlly higher, your calculator either converts microns to furlongs-per-fortnight or your batteries need replacing. When occasion arises where industry standard parts can be used, we use them. Its less expensive and it saves our company money as well. In light of the life of a microscope and the service required over that course of that period, there are times when you eat the bear or the bear eats you. In most cases, the service records will reflect that at the time a scope is retired, even at manufacturer cost, the contract customer still comes out ahead.
} "In a system as complex as an EM, manufacturer service engineers } seldom carry with them replacement modules. While they can order } such parts for rapid delivery, anyone can order those parts for } similar delivery - including you or a third party service source."
I think here you will find that the price you pay for your part will be at list price while the contract customer will not feel that sting.
Admittedly, your statement put me on the defensive. EM service engineers, no matter what company they work for, take considerable pride in what they do even though we may work on different sides of the market share table.
Respectfully,
Clay Jordan District Customer Service Engineer FEI/Philips Electron Optics
Well, we all have to do our part...I had a few minutes to kill, so I did a little investigation on this particular "justamateurs.com" website, AKA "208.153.103.7".
A "traceroute" revealed the gateway and the route through the 'Net for these spammers. I e-mailed and complained to their upstream providers. Who knows, it might make a difference. But then again...
Do your part to fight Spam!
Bob ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Leica (Wild, Leitz, Bausch&Lomb, Cambridge, AO, Reichert-Jung) / Technical Instrument Company / American Volpi / Fostek / Stocker and Yale / AEI North America / OptiQuip / Dolan-Jenner / Osram / G.E. / Philips / Ushio / Boeckler Instruments / Heidenhain / Narishige / Colorado Video / Visual Environments of California, Inc. / Kinetic Systems / Pacific Precision Laboratories, Inc. / Pryor Scientific / Compumotor / Sutter Instrument Co. / Advanced Database Systems / Cohu / Javeline Electronics / Optronics / Diagnostic Instruments, Inc. / Dage MTI / Hitachi / Panasonic / Polaroid / Kodak / Mitsubishi / Sony
} Attachment converted: Macintosh HD:Compress (TEXT/MSWD) (0000912D)
I would just like to appeal to everybody in this mailserver not to use attachments! It is one thing to delete a mail that is not within the scope of own interest but quite another thing to fiddle out all unwanted attachments from attachment folders. For longer statements please use personal Email! Thanks to everybody Hiltrud Mueller-Sigmund
Dr. Hiltrud Mueller-Sigmund Institut fuer Mineralogie, Petrologie und Geochemie Albertstrasse 23b, 79104 Freiburg (Germany) Tel.: (+49)-203-6388/-6396 Fax: -6407
I'm from Germany, Zip-drives are not such widely distributed here compared to the states, so they are no "must" for us (except people from the states come here with their data ...). I have nothing against ZIP-drives, I only want to ask why people state that they are cheap and handy for image transfer? In Germany they cost } = 15 US-Dollars in a normal Computer shop. For my feeling this is not cheap for 100 MB. Most media are cheaper (and faster), including MOs, CD-RW, Hard Discs (!) etc. Or do you pay only 5$ in the states?
Dr. Arthur Schuessler University of Heidelberg Zellenlehre Im Neuenheimer Feld 230 D-69120 Heidelberg Germany
We have a JEOL 1200II STEM and are considering the purchase of a double = tilt holder for SAED.
Does anyone have experience with the JEOL double-tilt, or is there a = better choice?? It's a very expensive item for a very specific = technique, and I have no personal experience.
Thanks,
Nan Laudenslager Analytical Testing Group Specialty Minerals, Inc.
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.2106.6"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} We have a JEOL 1200II STEM and are = considering=20 the purchase of a double tilt holder for SAED. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Does anyone have experience with the = JEOL=20 double-tilt, or is there a better choice?? It's a very expensive item = for a very=20 specific technique, and I have no personal experience. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Thanks, {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Nan Laudenslager {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Analytical Testing = Group {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Specialty Minerals,=20 Inc. {/FONT} {/DIV} {/BODY} {/HTML}
14th International Congress on Electron Microscopy
Cancun, Mexico
August 31 to September 4 1998
IMPORTANT NOTICE
The deadline for receipt of abstracts has been extended to
February 28
If you need further information please feel free to contact me - or to consult the Congress web site which has full details: http://icem.inin.mx . Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Vin Berry vinod.berry-at-gep.ge.com } ---------- } From: shAf[SMTP:mshaf-at-darkwing.uoregon.edu] } Sent: Thursday, February 12, 1998 4:21 PM } To: Microscopy Listserver; Nestor } Subject: bulk ads on Microscopy list } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I just received bulk mail from } mailto:art_and_mikey-at-justamateurs.com. Has anyone else received the } sexsite advertisement??? Even if I was single out somehow, the } Microscopy list was responsible somehow because it had the Microscopy } text header ... } } } cheerios, shAf } -- } {\/} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {\/} } } Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ } Geological Science's Electron Probe Facility at the University of Oregon } } mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu } }
We have a PSEM 501 which has a couple of HT problems (needs new HT cascade generator and gun silicone potting needs replacing). However, we are replacing it shortly and do not wish to spend any more money on it. It is thus available for spares to those of you still running them. SED, BSED, some signal processing, alpha numeric generator etc. Any parts left will be disposed of at the end of February.
Ron
========================================================================== = Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ========================================================================== ==
1. Zip disks are typically available for $15 or less as singles, but if you purchase in 6-packs or 10-packs, they are in the $10-12 range, and sometimes discounted even more. Iomega-brand Zips command the highest prices, but Sony and Maxell and Fuji also are licensed suppliers whose co-branded disks typically are a bit lower priced.
2. I believe that Zip drive data transfer rates are a *lot* faster than any optical drive, but of course do not match hard drive speeds. Check direct comparison reviews such as in PC Magazine, June 1996 for numbers. However, they are "fast enough", and can run many commonly used programs directly from the disk with ease, giving hardly any feel of running slower than from a hard disk. In fact, one of my colleagues had a hard drive failure some time ago, and ran most of her programs such as Word and QuarkExpress directly from the Zip for a long time.
3. I don't know about Germany (but hope to find out personally when I visit in April :-) ) but in the US you can take a Zip disk into any Kinko's Copies, for example, and be assured that you will be able to use their Zip drives to read your disk. You could do that with a CD also, of course, but probably not with assurance using any other type of removable media.
4. We have a CD writer in our lab, and in two years as far as I am aware, not a single user has requested to take their data home on a CD...these days virtually everybody comes with Zips because they are so handy to be able to dump images on without the hassles of CD writers. However, we do all of our permanent image archiving on CDs. So everything has its price and its advantages and disadvantages....
Larry
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Fellow microscopists, Can anyone recommend a general plant cytochemistry book/monograph? TIA Hank Adams Electron Microscopy Lab New Mexico State University Las Cruces,NM 88003 phone: 505-6463600 fax: 505-6465665
Regarding the now ongoing debate I will side 100% with Allen. I have witnessed everything he describes first hand. My colleagues at this and at other institutions have had virtually identical experiences. I find it interesting that all of us have had these experiences with a company that shares the same name as Mr. Jordan's e-mail address. I have not deleted any of the remarks so others can compare with their own experiences.
Clay Jordan wrote: } } Allen, } } No disrespect intended, but I can't let this go by unaddressed. My } apologies in advance, Nestor. } } You state: } } "Manufacturer service: } } Generally considers providing service a necessary evil. Service } } personnel are not given an upward migration path,..." } } It appears that to you there is no "upward migration path", which I } assume is to mean some nebulous region of importance in one's mind, } and that results in a better paycheck. Due to the complexity of EMs, } service engineers cannot help but constantly increase their skills and } expertise. Our "upward mobility" is always in motion. It doesn't } stagnate and the permutation of service problems are infinite, } challenging, and enlightening...and it, too, results in better } paychecks. } } } "...thus leading to high turnover and a generally low rate of } } service engineer competency." } } Of the 70+ service engineers in the company I work for, the average } length of time the company is 10 to 15 years with several 25+ years } service, the least 3 1/2 years. Turnover is very rare and to my } knowledge has never been due to the lack of upward mobility. I will } not take issue with your statment that there is a 'generally low rate } of service engineer competency'. I assume you hire the best...so do } we. AND manufacturer service engineers are constantly being trained } and updated on any EM service modifications and new products. } } } "Part charges are generally 1000 - 1500 percent of the market value } } (99% of instrument problems require the replacement of electronic } } industry standard parts)." } } Though the manufacturer list price to non-contract customers is } significantlly higher, your calculator either converts microns to } furlongs-per-fortnight or your batteries need replacing. When occasion } arises where industry standard parts can be used, we use them. Its } less expensive and it saves our company money as well. In light of the } life of a microscope and the service required over that course of that } period, there are times when you eat the bear or the bear eats you. In } most cases, the service records will reflect that at the time a scope } is retired, even at manufacturer cost, the contract customer still } comes out ahead. } } } "In a system as complex as an EM, manufacturer service engineers } } seldom carry with them replacement modules. While they can order } } such parts for rapid delivery, anyone can order those parts for } } similar delivery - including you or a third party service source." } } } I think here you will find that the price you pay for your part will } be at list price while the contract customer will not feel that sting. } } Admittedly, your statement put me on the defensive. EM service } engineers, no matter what company they work for, take considerable } pride in what they do even though we may work on different sides of } the market share table. } } Respectfully, } Clay Jordan } District Customer Service Engineer } FEI/Philips Electron Optics } "OK...Who put Stop-Leak in the water chiller?"
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
I am using a fluorescence scope to examine whole mounts of unfixed leaf material, when using a triple excitation filter #83000 for DAPI/FITC/TR we have been finding that the leaf material will heat up to a point that charring of the specimen is observed. This is with the standard heat absorbing glass filters on an Olympus BX60 in place. Could this be some unblocked leakage of IR with this filter set? As it is not seen when using single band excitation for each fluorochome separately.
If this is the fact would it dangerous (from the point of Hg lamp brakage) to install a gold hot mirror filter to block to IR above 650 nm.
Thanks
Russ Spear Russell N. Spear Sr. Research Specialist Dept. of Plant Pathology Univ. of Wisconsin-Madison
Just to keep a little perspective here, I must say that my experience does not coincide with Geoff's. At least not in the case of Mr. Jordan's company. During my tenure as a facility administrator I have owned TEM's from 4 manufacturers. While I have had some problems with the service from 1 of the manufacturer's, I have never had any problems with Mr. Jordan's company and I have operated a 410LS since 1983. During that time I have dealt with the same two service engineers. My experience is shared with another facility on campus with three more of their devices. Indeed, it is the quality of their service that allows me to recommend their company over any of the others I have dealt with. Sure, my experience is somewhat anecdotal, but so is everyone's. I cannot agree with Allen or Geoff.
Respectfully,
Rick A. Harris, Director Microscopy and Image Analysis Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 752 3085 fax raharris-at-ucdavis.edu
Enroll us in your lobby. And, space permitting, I'd like to publish them in the next issue of The Microscope Book. Regards, Elinor Solit, The Cambrex Group
On Thu, 12 Feb 1998, Caroline Schooley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } Is anyone else interested in lobbying to have Caroline's microscopy quotes } } list posted to the group? If it isn't too much trouble, of course - } } but some that were sent to the list have been wonderful (Woody Allen's, } } especially)...Would anyone be offended at the "wasted' bandwidth? } } } } Tamara Howard } } CSHL } } Plan B: They're so good that perhaps they belong in the MICRO section of } the MSA web page; want to look at them all, Nestor? } } Caroline } } } Caroline Schooley } Educational Outreach Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/ } }
The cost of the drive itself is also an important consideration as is=20 the wide acceptance of the Zip=2E =20 =20 John Humenansky Braun Intertec Minneapolis, MN =20 fax: 612-942-4844
------------------------------------------------------------------------=20 The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20 =20 =20 I'm from Germany, Zip-drives are not such widely distributed here compared=20 to the states, so they are no "must" for us (except people from the states=20 come here with their data =2E=2E=2E)=2E I have nothing against ZIP-drives, I only want to ask why people state that= =20 they are cheap and handy for image transfer? In Germany they cost } =3D 15=20 US-Dollars in a normal Computer shop=2E For my feeling this is not cheap fo= r=20 100 MB=2E Most media are cheaper (and faster), including MOs, CD-RW, Hard=20 Discs (!) etc=2E Or do you pay only 5$ in the states? =20 =20 Dr=2E Arthur Schuessler University of Heidelberg Zellenlehre Im Neuenheimer Feld 230 D-69120 Heidelberg Germany =20 Fax: 06221 544913 =20
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I just had another thought about Zip drives that have not been covered in the previous conversations about portable media.
I don't want a large capacity portable disk to carry around my data and working files. I have the option here of doing this with 1Gb Jaz drives and still choose the Zip drives. Most times, I only have a few images in the 1-2 Mb range with Powerpoint files for presentations that are in the 10-30 Mb range and perhaps some spectra and a WordPerfect file. I can label this disk with the topic that it covers and carry it home and work with it there. Ok, sometimes I need two Zip disks, but two of them can still fit in my pocket easily. I consider the Zip disks my working disks. When the information is ready for archiving, it gets burned onto a CD.
If I have too much stuff on one disk, I forget what's on it and can't keep up with it. I also think that I run too much of a risk if something happens to it. My mode of operation is to use the Zip for the current file and backup the file on the hard drive of the last machine that I used. This way, if I loose the Zip disk, I can still go back to the last machine that I used.
Just my two cents.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Hank Adams wrote: ============================================ Fellow microscopists, Can anyone recommend a general plant cytochemistry book/monograph? ============================================= One book that is a very good seller, is the following:
You can get an over view of the book and the complete Table of Contents listing on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Hi! I received a sample of fungus (potato blight) grown on an agar, with the request to process it for SEM. Does anybody know the best way to aproach it? Thanks Dorota e-mail Wadowska-at-UPEI.ca
Dear Hank, an excellent but almost impossible book to procure (it's out of print) is:
O'Brien, T.P. and McCully, M.E. (1981) The study of plant structure: Principles and selected methods. Termarcarphi Printing Ltd., Melbourne. ISBN 0 9594174 0 0
Also good is:
Berlyn, G.P. and Miksche, J.P. (1976) Botanical microtechnique and cytochemistry. Iowa State University Press. ISBN 8138 0220 2
Both a little dated (is there a later edition of Berlyn and Miksche?) but very good for basic techniques. Cheers, John
================= C. John Runions Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
Note also Research post doc position at ASU. Contact Lindsay lab http://green.la.asu.edu/
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Nestor, my apologies in advance for continuing this digression, however, some out there may be interested in this behind-the-scenes look.
} Allen, } } No disrespect intended, but I can't let this go by unaddressed. } My apologies in advance, Nestor. } } You state: } } "Manufacturer service: } } Generally considers providing service a necessary evil. } Service } personnel are not given an upward migration } path,..." } } It appears that to you there is no "upward migration path", } which I assume is to mean some nebulous region of importance in } one's mind, and that results in a better paycheck. Due to the } complexity of EMs, service engineers cannot help but constantly } increase their skills and expertise. Our "upward mobility" is } always in motion. It doesn't stagnate and the permutation of } service problems are infinite, challenging, and } enlightening...and it, too, results in better paychecks.
Nothing nebulous about it. Young people entering a field like to find a horizon beyond their current position. Yes, a field engineer's salary will increase over the years, as will any profession. The problem generally relates to the first statement above, that manufacturers consider service a necessary evil.
Few manufacturers really recognize the benefits of a good service organization. To corporate management, it is a pesky drain on corporate profits. To engineering, it is something that will eventually be engineered out of the picture. While I put this in rather dire terms, there are a few very revealing questions I could pose to any manufacturer. How many corporate management positions have been filled by former field engineers? More to the point, how much training have your field engineers been given in cross selling your products?
} } "...thus leading to high turnover and a generally low rate of } } service engineer competency." } } Of the 70+ service engineers in the company I work for, the } average length of time the company is 10 to 15 years with } several 25+ years service, the least 3 1/2 years. Turnover is } very rare and to my knowledge has never been due to the lack of } upward mobility. I will not take issue with your statment that } there is a 'generally low rate of service engineer competency'. } I assume you hire the best...so do we. AND manufacturer service } engineers are constantly being trained and updated on any EM } service modifications and new products.
My statements were intentionally broad - there are some good companies out there. However, there are far more that can not maintain a competant service force and service presence. I can attest to Philips service as generally being found adequate by customers, since I have only infrequently been requested to provide service on your instruments (however, since I primarily work on SEMs, perhaps that relates more to your market share in that area).
} } "Part charges are generally 1000 - 1500 percent of the market } value } (99% of instrument problems require the replacement of } electronic } industry standard parts)." } } } Though the manufacturer list price to non-contract customers is } significantlly higher, your calculator either converts microns } to furlongs-per-fortnight or your batteries need replacing. } When occasion arises where industry standard parts can be used, } we use them. Its less expensive and it saves our company money } as well. In light of the life of a microscope and the service } required over that course of that period, there are times when } you eat the bear or the bear eats you. In most cases, the } service records will reflect that at the time a scope is } retired, even at manufacturer cost, the contract customer still } comes out ahead.
I could provide an endless list of remarkable part charges by nmanufacturers. Most companies serve parts from their own storerooms where part costs include a very healthy margin for the procurement and storage. I will also add that some manufacturers modify third party assemblies or software to lock customers into them as a sole source.
Here's a simple example from just a few years ago. The venerable DEC LSI-11 used in EDS was generally assembled from a variety of suppliers. Seldom was the customer supplied with the complete operating system, often the system utilities required for complete maintenance were intentionally left out. I had a customer who had a 40MByte hard drive go bad who was quoted $6000 for its replacement. The manufacturer felt safe in asking this price because they had made a slight modification to the driver board and had not supplied the hard drive formatting utility with the system.
In this case, a simple call to the manufacturer of the driver board got us a free copy of the formatting utility and the drive was found through a number of sources for $400.
Your reply does raise a question, though. Are you stating that Philips charges more for parts to non-contract customers than it does to contract customers? Is this legal?
Yes, Philips can easily claim that contract customers come out ahead. I have seen the labor rates you charge for your billable services.
} } "In a system as complex as an EM, manufacturer service } engineers } seldom carry with them replacement modules. } While they can order } such parts for rapid delivery, anyone } can order those parts for } similar delivery - including } you or a third party service source." } } } I think here you will find that the price you pay for your part } will be at list price while the contract customer will not feel } that sting.
Once again, is this practice legal? While there are certain games you can play to entice customers into contracts such as response time and labor rates, I don't think that you would be on good legal ground charging different part prices. In dealing with manufacturers for part orders, I have never even been asked if I own one of their instruments, much less whether I have one under contract. But then, I have obviously not had to order a Philips part.
} Admittedly, your statement put me on the defensive. EM service } engineers, no matter what company they work for, take } considerable pride in what they do even though we may work on } different sides of the market share table.
True enough, for those who find their niche here. But there are obviously still some very serious differences of opinion on customer service.
} Respectfully, } } Clay Jordan } District Customer Service Engineer } FEI/Philips Electron Optics
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
I have tried examining Ca metal that had been mounted and polished. The = tricky part is getting the mount into the SEM before the surface = oxidizes enough to interfere.
I thought about adding something to the last polishing felt to seal the = surface. Maybe a formvar solution?? Any other thoughts??
Thanks,
Nan Laudenslager Analytical Testing Group Specialty Minerals, Inc. nhl-at-early.com
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.2106.6"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I have tried examining Ca metal that = had been=20 mounted and polished. The tricky part is getting the mount into the SEM = before=20 the surface oxidizes enough to interfere. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} I thought about adding something to = the last=20 polishing felt to seal the surface. Maybe a formvar solution?? Any other =
Dorota, I would take plug-like samples of the fungi and the agar below it using a widened pipette tip place the samples into a small plastic petri dish along with a beem capsule lid containing 1 or 2% osmium tetroxide (in buffer if you wish). Seal and insert this into a larger petri dish. Seal and place the nested dishes in a dark box in a fume hood for 48 hours. Examine every 12-15 hours and replace osmium with fresh soln. The fungi should turn black and the agar only a pale grey. Remove fixative to a waste container with vegetable oil and allow the sample to air dry for 24 hours. Trim off the agar, glue the disk of fungi to an aluminum stub with double sticky tape, add colloidal silver to the edge of the sample and tape. Sputter-coat with 10 nm of gold or gold/palladium. If the fungi has loose threads or fruiting bodies, I would prepare another stub with double sticky tape and touch it to the surface of another plug to obtained structures fully adhering to the stub-- again add a drop of colloidal silver and sputter-coat. This is known as vapor fixation and is very effective for hydophobic organisms such as fungi. I do not have a reference handy but can look for it at the lab on Monday. Good luck! Rosemary
At 3:33 PM 2/13/98 -0400, Dorota Wadowska wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
At 01:36 AM 2/14/1998 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to toss in my experiences regarding EM service contracts. We had our Brand X SEM on a service contract for several years. The manufacturer promoted an excellent service engineer to another position and region. His replacement was not properly trained on our instrument and could not handle the volume of instruments entrusted to his care. Consequently, our instrument began to suffer and complaints were not dealt with constructively. Other local users of this brand had similar frustrations.
I spent a lot of time and energy persuading the manufacturer to correct their problems and compensate us for their transgressions. After an extension of the contract expired, I searched for a third party service provider and found....Allen Sampson. I can certainly vouch that, while I am not contracted for next-day service, the technical competence is at least as good as, if not better than the manufacturer's. Additionally, we purchase standard replacement parts for a fraction of the list price.
While I have to assume some of the maintenance decisions myself and be responsible for obtaining replacement parts, the instrument is running just fine. This situation may not be for everyone, but it is working out very well for me now. Perhaps if Brand X would have retained the original service engineer or not had so many organizational overhauls, I might still be on their service contract.
We had another situation some time where the service oragnaization was so atrocious, the local service engineer (good and competent) was so fed up with his company that he quit as our system was being installed. The system never performed as the manufacturer quoted, and they flat out said that they would not honor their quote. After eight months of fighting with them, their system was removed to our shipping dock for them to retrieve.
The point is that there are good and bad vendors. As consumers of high ticket items we need to protect ourselves by written contract and sharing information about errant vendors (within legal/ethical boundaries). I would like to believe that there would be an uncompromising level of professionalism by the vendors, given the relatively small niche market and risk of a bad reputation. Forums like this can help us all by exposing us to different opinions, options and experiences.
Regards,
Alan Stone
I am not affiliated with Allen Sampson in any manner other than being a service client. This is an unsolicited posting without compensation. Alan Stone ASTON Metallurgical Services Chicago
While I don't want to get embroiled in heated discussions about service contracts, I will comment briefly on my own experience. The value of a service contract depends largely on the manufacturer and their service for your particular geographical location. I have had a service contract with Amray which has been well worth the money - great service engineers who have been around for as long as the contract has (15 years) and a very quick response time. But I have also had contracts with other manufacturers (who shall remain nameless) which were not worth anything - incompetent service, poor phone support, constantly changing personnel, long wait times for service, etc... At the risk of stating the obvious, it makes sense to evaluate the manufacturer service during the first year of service and then make a decision as to whether to continue or find a third party. Good service can promote future sales of instrumentation and bad service can prevent future sales.
SEM/EMP I am examining geological thin sections with nucleic acid stains at high magnification with an oil immersion objective. Subsequently, I wish to examine these samples with the SEM/EMP. The question I prose is what is the best way to remove the immersion oil so that I can examine this sample under an electron beam?
Ken Tobin Guyot Hall Dept. of Geosciences Princeton University Princeton, NJ 08544 Ph: 609-258-1383 FAX: 609-258-1274 e-mail: tobin-at-geo.princeton.edu
Literary microscopists: There has been amazing response to my request for quotes for the MSA/LHS middle school microscopy manual; I've received a lot directly, in addition to the ones that have appeared on this listserver. Many of you have requested copies, but it's too large a file to post here (2100 words, 7 pages!). So here's the solution: They'll appear at intervals as "fillers" in the MSA Bulletin, and I'm asking Nestor to post the whole thing in the MICRO section of the MSA web page; I'll let the listserver know when it's there. Those of you who contributed did so for nonprofit educational purposes; this added use is in the same spirit, but if you don't want your contribution used, please let me know. And if you have "just one more", send it on!
Thank you for the lovely Valentine.... Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Although it is true that the initials cost of MO media is high (about US$10 for 230 MB of reusable space) the gain in stability and safety make it a superior media for archiving data.
It is categorically untrue that MO disks are subject to stray magnetic fields, as some of you have stated. While they ARE magnetic media, the particles are embedded in polymer that must be heated to 700 degrees by the laser before they can be realigned. You could place one of the disks in a car, then pick up the car with a giant electro-magnet at the junk yard--and your data would be safe! In fact, data stored in this way is guaranteed for a minimum of 30 years.
In our facility, flow cytometric data has been stored in this way for many years, on a variety of devices with no failure of machine or media. Just treat the machine right! Don't leave media in the machine all the time. ( This invites dust). Remove the media before moving the machina around.
In addition MO is faster than CD-ROM with sustained transfer rate of 2MB/second on 540 and 640 MB diskettes and faster yet on direct over write (LIMDOW) capable machines and media.
later -gene
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
The Boulder Laboratory for High Voltage Electron Microscopy seeks a person to maintain and improve its 1,000KV electron microscope and associated equipment. Job will include research on instrumentation for high resolution imaging of cells. Experience with electronics required. Experience with the following is desirable: repairing electron microscopes and high vacuum systems, UNIX systems administration, C programming, and implementing computer control of equipment. Salary $40,000 to $55,000, depending on experience. Send resume and references by Feb. 20 to J.R. McIntosh, Univ. Colorado, Boulder, CO 80309-0347.
The University of Colorado is committed to diversity and equality in education and employment.
Quite often, the MO disk formated from one drive could not be read from another drive (with another brand). Any ideal how to deal with such problem?
Thanks
gene a s wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } To All: } } Although it is true that the initials cost of MO media is high (about } US$10 for 230 MB of reusable space) the gain in stability and safety } make it a superior media for archiving data. } } It is categorically untrue that MO disks are subject to stray magnetic } fields, as some of you have stated. While they ARE magnetic media, the } particles are embedded in polymer that must be heated to 700 degrees by } the laser before they can be realigned. You could place one of the disks } in a car, then pick up the car with a giant electro-magnet at the junk } yard--and your data would be safe! In fact, data stored in this way is } guaranteed for a minimum of 30 years. } } In our facility, flow cytometric data has been stored in this way for } many years, on a variety of devices with no failure of machine or media. } Just treat the machine right! Don't leave media in the machine all the } time. ( This invites dust). Remove the media before moving the machina } around. } } In addition MO is faster than CD-ROM with sustained transfer rate of } 2MB/second on 540 and 640 MB diskettes and faster yet on direct over } write (LIMDOW) capable machines and media. } } later } -gene } } _____________________________________________________________________ } You don't need to buy Internet access to use free Internet e-mail. } Get completely free e-mail from Juno at http://www.juno.com } Or call Juno at (800) 654-JUNO [654-5866]
-- ******************************************************** Dr. Yifan Cheng Internatioanl Institute for Advanced Research Central Research Laboratories Matsushita Electric Industrial Co., Ltd. 3-4 Hikaridai, Seika, Kyoto 619-02, Japan
please take care when exchanging the TEM film magazines. Inserting the new magazines slightly misaligned could result in a blocking of the transport mechanism (like last Friday)!
This usually causes more or less intensive service work!
Thanks to all the people who suggested manufacturers that could provide me with a periodic table with X-ray data printed on it. Suggestions included (in no particular order): EDAX Noran Oxford Instruments Philips Kevex Princeton-Gamma-Tech In the end, both Noran and EDAX sent me copies of their periodic tables in the post so I now have what I need.
Thanks again everyone
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
To those using Dr. John Russ's Image Processing Toolkit, I have a question...
Has anyone tried using the FILTER / IP*MEASURE / GLOBAL selection? When I do, the Area Fraction is reported but not the total image area. I am trying to find a simple way to measure the total image area (calibrated) as a reference for other measurements (count and area fraction), but can't seem to do it.
I am using version 2.1.0 running under Windows 95 and Adobe Photoshop 4.0. I have dowloaded the updates up to #04.
BTW, I think these Plug-Ins are wonderful! I've been using them for about 6 months, off and on, and this is my first stumbling block. Thanks in advance for any suggestions, Karen
-- Karen Zaruba, Life Sciences Sector Laboratory, 3M Company, St. Paul, MN 55144 kszaruba-at-mmm.com
*The opinions above are my own, not necessarily my employer's*
Very soon, in fact on wednesday, I have to fix some bone marrow for EM epon and lowicryl embedding. I do not know if the sample comes from spine aspirates or from a hip puncture. Any help about the best way to fix this material would be really appreciated, Thank you Daniele -- =
************************************************************** * Dani=E8le SPEHNER, CJF 94-03 INSERM * * Etablissement de Transfusion Sanguine de STRASBOURG * * 10 rue Spielmann, B.P. 36- 67065 Strasbourg-Cedex, FRANCE. * * Tel : (33) 03 88 21 25 25 - Fax : (33) 03 88 21 25 21 * **************************************************************
{/bigger} Double-tilt holders are essential for Materials Science applications. Perhaps you can buy a used holder off of the list server. I would also look into a Gatan holder (if available). Replacement hex-type screws for the JEOL are very pricey about $800.00 and they are easy to lose. Good luck !!
I found some treasures in the lab for a microscope we don't have anymore. Phillips I think? We had the 300 in the past, but since it's long gone, these items are of no use to me. Maybe someone out there has a need?
Items: 6 film boxes for Phillips, about 3 7/8" X 4 1/4" X 2 3/4", complete with the film plates, 4 are supply, 2 are receivers
O-ring set for the same instrument: part # like R208, SOR104, R4118, R4137/MS28775-2201/PO2563, etc. you get the idea... unused in zip-lock plastic bags.
First come, first serve!
************************************************************ It's true- the inmates ARE running the asylum... ************************************************************ Laura Rhoads Electron Microscopy Facility Director Department of Biology Western Kentucky University 1 Big Red Way Bowling Green, KY 42101-3576
} Has anyone tried using the FILTER / IP*MEASURE / GLOBAL selection? When } I do, the Area Fraction is reported but not the total image area. I am } trying to find a simple way to measure the total image area (calibrated) } as a reference for other measurements (count and area fraction), but } can't seem to do it.
Good suggestion- it will be incorporated in the next release... (and thanks for the kind words) John Russ
Service should be a MAJOR consideration in the decision to purchase an instrument. The extra bells and whistles that swayed you towards a particular brand aren't worth much later when your scope is down for other reasons.
Also, this thread began with a survey of service contract options. One overlooked option is doing some of the service yourself and taking advantage of discount service contracts offered by most manufacturers. By doing some of your own repairs, you may 1) save some money, 2) learn more about your instrument, 3) develop some empathy for field service engineers (who aren't getting much respect in this discussion).
Owen
Disclaimer: My employer hired me because of the views expressed in this message.
============================= Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
The microbe is so very small You cannot make him out at all, But many sanguine people hope To see him through a microscope. His jointed tongue that lies beneath A hundred curious rows of teeth; His seven tufted tails with lots Of lovely pink and purple spots, On each of which a pattern stands, Composed of forty separate bands; His eyebrows of a tender green; All these have never yet been seen - But scientists,who ought to know, Assure us that they must be so ... Oh. Let us never, never doubt What nobody is sure about.
Hilaire Belloc (probably)
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney AUSTRALIA 2001
There's another thing that manufacturers are up against and so are we independents, but not usually to the same degree. Look at the job description:
You want someone who is technically competent, not just with theory, but with real-world horse-sense. And competent not only in both digital and analog electronics, but vacuum, precision mechanics, particle physics, x-ray chemistry, computer technology, software, and..."oh, it would be nice if you could talk intelligently to your customers about their field of interest and help them with their applications problems in materials, microelectronics, geology, biology, medicine, forensics, and failure analysis, just to name a few".
You want someone who can work alone day after day after week after year. You want someone who is willing to work out of a suitcase most of the time but you really want the maturity and stability of a married person with a family. You want someone who is stubborn (ornery?) enough to stick with any problem he comes across, including PEBCAK (problem exists between chair and keyboard). You want someone who can deal pleasantly with all customers, good and bad. You want someone with the self motivation to actually go to work each day and put their best into it without any supervision. You want someone to whom you can give a corporate AMEX card, company car, and $5-15,000 (or more) worth of tools and parts and not worry about getting ripped off.
Trying to find any 2 or maybe 3 of these in one person can sometimes be a job! Trying to find them all is nearly impossible because many of the requirements tend towards being mutually exclusive in most people.
The manufacturers are really up against it because they have a clearly defined and growing customer base. They HAVE TO have enough warm bodies out in the field even if they don't all meet spec. I remember as eastern service manager knowing who I needed to fire, but needing the warm body to be able to send even more.
Alan, you and I worked for a damned good company before it was Perkin-Elmered, but even ETEC had poor engineers and the occassional thief. You are also right that service personnel don't generally make it to upper management. It's a terrible loss to these companies because the focus of any customer service person worth his/her salt is solidly on the customer (not the potential customer, shareholder, etc.). They know what the business is as no MBA could ever begin to comprehend.
We have the advantage of sizing our companies to the number of properly skilled people we have, if we want to. We can also say "No, I don't care to do business with you." for what ever reason we deem sufficient. The manufacturers don't really have those luxuries.
The person that shows up on site is the most important part of the puzzle. This was shown to me when I started Quality Images. Many people were willing to give up the security of being serviced by the manufacturer, which was still in business at the time, to be certain who would show up on site.
Yes, there are excellent field engineers out there working for manufacturers, some of whom support their field staff better or worse than others. My hat is off to those who like this work and put their all into it. But then I can say that about virtually any person in any occupation who puts their all into it.
Alan and Clay, I don't have to tell you that this is a tough business to be in in some ways, but I don't think all of the users appreciate that it's tough. That said, you also know that it can be one of the most satisfying jobs there is and a fair portion of the users are just wonderful people to hang around with.
As for pricing.....This business is expensive. ETEC's mark-ups were huge, to my way of thinking, and at the time, their contracts were among the most expensive, but they had to be bought because they were broke (for reasons other than service, but broke just the same). We can restrain our service area, if we choose. The manufacturers can't, unless they restrain their sales area (decidedly unlikely for an international company). By virtue of their size, they have far more overhead, but they have access to all the parts, all the documentation, all the software, all the source code and the largest group of people qualified to work on their equipment.
Given all that, they still manage to alienate some people sufficiently to cause them to risk it all for a different person on site. There have also been independent service companies that have made confirmed manufacturer fans of some users.
There is room for all of us. Not every user's needs are the same, just as not every engineer's mode of operation is the same. Some of us, who are truly loved at some sites, can be truly disliked at others. Some people swear by their instrument, others swear at the same model. Who's right and who's wrong?
To the users, take this series of letters with a grain or two of salt because if you don't already know, I'll tell you a little secret...... field service engineers tend to be very opinionated, myself included.
As to whether you should have a contract and who you should have it with, the pros and cons have been pretty well covered by others. Everything is a trade-off. It's like running your SEM.....do you want high magnification or good depth of field with lots of signal? You have to make a decision. Hopefully your decision will suit your needs. That is the bottom line.
Ken Converse owner Quality Images 16 Creek Rd. Delta, PA 17314 717-456-5491 717-456-7996 fax qualityimages-at-netrax.net
Some of you may remember our old friend the HODJA (or Mullah) Nasr-ed-Din. Here are two more stories about him.
* * * * * * * * * * * * *
It was a cold winter night, clear and frosty with a full moon. At about 10:30, a friend came to visit the Hodja, and found him outside his house crawling around on hands and knees.
"What are you doing?" asked the friend.
"I've dropped a coin, and I'm looking for it" replied the Hodja.
"May I help you search?" offered the friend.
And so the two men crawled around together for over an hour, until it was around midnight. The friend was starting to feel frustrated.
"Don't you have any idea where you dropped it?" he asked.
"Oh yes," replied the Hodja, "in the bedroom."
"Then why aren't we looking there?"
"Because it's dark in the bedroom, and there's a full moon out here!"
At one phase in my life as a scientist (!?) I was involved in a succession of three-month contracts funded by Company C. Their materials were mainly in the form of thin films, and in order to do a cross-sectional examination, I had to sandwich them between thicker layers using a certain polymeric "butter". Of course, supplier S had it in stock, and delivery would be a week at most. Six weeks in fact, so in the meantime I had to direct my research somewhere else, like the Hodja searching for his coin.
* * * * * * * * * * * * *
The Hodja once came home at lunchtime, carrying 3 pounds of liver, which he handed to his wife, to prepare for his dinner. But during the afternoon, some of her friends came round, so she cooked it and they all ate it together. In the evening, the Hodja came home and found his expected dinner missing.
"Wife, where's the liver I brought home for dinner!" he roared.
Suddenly his wife spotted the cat, and had an idea.
"The cat ate it!" she cried.
"Oh really?" said the Hodja "we'll soon see about that".
He picked up a pair of scales, and put the cat on it. It weighed exactly 3 pounds.
"There you are!" cried his wife triumphantly.
The Hodja looked puzzled for an instant, then asked:
"3 pounds, yes. But if this is the cat, where is the meat? And if this is the meat, where is the cat?"
Sometimes the supplier of an end product comes along with two specimens, and asks, why has A failed, and B not? Alas, when one comes to study the situation, one finds that A is made from "Crummilon" and B from "Tackynar", i.e. two competing grades of the same material (or even worse, and old and new version the same material), but A was moulded by procedure X, and B was extruded by procedure Y, then A was used in environment 1 and B in environment 2. And so forth ...
And there is political pressure from various quarters to blame the original material, the production method, the user, whatever or whoever.
One subjects A and B to one's state-of-the-art morphological analysis, and does find some differences. But where does one put the blame? One can suggest answers, but one generally has not enough information to choose between meat and cat.
* * * * * * * * * * * * *
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
If you are running Win 3.1, this seems to be a normal "feature" {g} . Less likely with W95. In 3.1, sometimes two different programs/drivers can try to use the same memory locations - crash! Otherwise... The first other thing that comes to mind is flaky memory. This can be true even if boot-up checks fine. If available, try swapping for some different memory.
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Hi,
There is some sort of minor software problem with our Edax computer. A few times a day, during heavy use, the computer will declare a general protection error. The only way out of this error is to close all programs and exit windows and then restart. After restart nothing appears to be strange, until several hours later another general protection error occurs. I have been looking for a pattern, but have been unsuccessful to date.
by vgernet.net with SMTP; 17 Feb 1998 14:48:20 -0000 Received: from web.vgernet.net (root-at-web.vgernet.net [205.219.186.3]) by photon.vgernet.net (8.7.6/8.7.3) with ESMTP id JAA18977 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 17 Feb 1998 09:48:19 -0500 Received: from spm11.vgernet.net (spm11.vgernet.net [206.25.237.111]) by web.vgernet.net (8.7.6/8.7.3) with SMTP id JAA10352 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 17 Feb 1998 09:48:15 -0500
Dear fellow microscopists,
I was really hoping that someone else would send this message, but no such luck {grin} .
As part of the exchange regarding service contrasts, comments were made about the high prices charged by manufacturers for service spares. The situation looks like this from the perspective of this manufacturer of specimen preparation equipment:
Our customers know that we manufacture most of the equipment which we sell. They expect that, when they need to replace a part, we will have the part in stock for immediate shipment, reducing their down time to a day or two at the worst. This means, in effect, that we need to inventory all of the parts and subassemblies for all of the instruments we build. In the case of a relatively complex table-top instrument, like the JB-4 microtome, this can be a few hundred line items.
As a result, we need to charge customers a sufficiently high price for the spare parts to cover our cost of maintaining this inventory. Our markup on spare parts is larger than our markup on the instruments themselves, and certainly larger than our margin on items which we simply order as needed for resale. The reason for this is not greed, but our company policy to provide the most rapid possible turnaround to customers who have purchased our equipment.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
} From {http://ciac.llnl.gov/ciac/CIACHoaxes.html}
Among mostly serious information it also contains the following spoof of the Good Times virus hoax that is just too good not to repost here, although it of course has nothing whatsomuchever to do with translation. Same could be said about Join the crew, I believe...
READ THIS:
Goodtimes will re-write your hard drive. Not only that, but it will scramble any disks that are even close to your computer. It will recalibrate your refrigerator's coolness setting so all your ice cream goes melty. It will demagnetize the strips on all your credit cards, screw up the tracking on your television and use subspace field harmonics to scratch any CD's you try to play.
It will give your ex-girlfriend your new phone number. It will mix Kool-aid into your fishtank. It will drink all your beer and leave its socks out on the coffee table when there's company coming over. It will put a dead kitten in the back pocket of your good suit pants and hide your car keys when you are late for work.
Goodtimes will make you fall in love with a penguin. It will give you nightmares about circus midgets. It will pour sugar in your gas tank and shave off both your eyebrows while dating your girlfriend behind your back and billing the dinner and hotel room to your Discover card.
It will seduce your grandmother. It does not matter if she is dead, such is the power of Goodtimes, it reaches out beyond the grave to sully those things we hold most dear.
It moves your car randomly around parking lots so you can't find it. It will kick your dog. It will leave libidinous messages on your boss's voice mail in your voice! It is insidious and subtle. It is dangerous and terrifying to behold. It is also a rather interesting shade of mauve.
Goodtimes will give you Dutch Elm disease. It will leave the toilet seat up. It will make a batch of Methanphedime in your bathtub and then leave bacon cooking on the stove while it goes out to chase gradeschoolers with your new snowblower.
Listen to me. Goodtimes does not exist.
It cannot do anything to you. But I can. I am sending this message to everyone in the world. Tell your friends, tell your family. If anyone else sends me another E-mail about this fake Goodtimes Virus, I will turn hating them into a religion. I will do things to them that would make a horsehead in your bed look like Easter Sunday brunch.
We have started using ProLong Anti-Fade from Molecular Probes and have been very pleased. However, CY-3 is the one I haven't tried it with.
Bob
On Fri, 13 Feb 1998, Patricia A. Glazebrook wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are trying to find supplier of citiflour anti-fade solution. Any other } suggestions for mounting CY3 specimens? } } Thank-you, } Pat Glazebrook }
} I don't know if this would help or not but it's a bit of a twist on the idea suggested by Caroline Schooley. When we mount our immunofluorescent slides we coverslip with mounting media as usual, but then we seal the edges with nail polish to prevent it from drying out. Just an idea . . .
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca } } } } I am new to this, both the internet and microscopy so if my questions are } } inappropriate for this forum, please excuse me and discard. } } } } My son Paul is 11 and received a good student microscopy for Christmas a } } year ago. He is an avid science student and is currently making slides of } } every feather he can find and is interested in his slides being more } } permanent. } } I have been trying to help him, but have not had much success. We started } } with the gum spirit in the Edmund Sci. microscope slide kit, but it takes } } forever to dry and seems to shrink as it dries causing terrible bubbles. I } } finally got some Cytoseal 60 locally and have had much more success however } } we still have problems with air bubbles creeping in from the edges days or } } weeks later. } } Are we using too much or not enough cytoseal? Do we need to seal the edges } } with something else? Is heating suggested to fully cure? How long should } } it take to dry? I thought this stuff was supposed to be fast, but it seems } } like days or weeks later the interior of the slide is still liquid. (or is } } it supposed to stay that way?) After drying should the specimen be soaked } } in solvent and if so which one? } } } } Any other tips for making simple slides would be appreciated. I've been } } hunting the net and found lots of good information, but not the answers to } } these. } } } } Thank you in advance and Paul also thanks you. } } } } David Bacque } } Proud father and mentor of an avid young scientist } } } } } } --------------------------------------------------------------------------- } } } } } } } Pat Hales McGill University Department of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
Can anyone verify that this is by Hilaire Belloc? Caroline } } THE MICROBE } } The microbe is so very small } You cannot make him out at all, } But many sanguine people hope } To see him through a microscope. } His jointed tongue that lies beneath } A hundred curious rows of teeth; } His seven tufted tails with lots } Of lovely pink and purple spots, } On each of which a pattern stands, } Composed of forty separate bands; } His eyebrows of a tender green; } All these have never yet been seen - } But scientists,who ought to know, } Assure us that they must be so ... } Oh. Let us never, never doubt } What nobody is sure about.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
A few months ago I posted a message asking for suggestions for a grade of stainless steel to use in making parts for a specimen holder rod for a side entry TEM stage. After discussing the matter with several metallurgists, and searching the literature for as much relevant info as I could find, I finally settled on trying grade 310 stainless steel, and am pleased indeed to report that it has worked out quite well. I used it to make the part that holds the specimen grid in a special rod for a 200 kV HRTEM, and there has been no sign of any magnetic interference, even at magnifications of 700,000.
Type 310 Stainless steel has a nominal composition of 24-26% Cr, 19-22% Ni, (with 0.25% max for carbon and 2.0 max for Mn), with the balance Fe. This is a somewhat higher level of chromium content than other common grades of StSteel(e.g. 302 has 17-19%, 316 has 16-18%, 304 has 18 to 20%, etc.), and this, according to my metallurgist friends, is what gives it its lower magnetic permeability. In any event, it worked out satisfactorily in this rather critical application, and so might be worth keeping in mind for anyone else who might get involved in a similar design problem.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
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You think that's something. We've had people put the TEM camera & the film holder in backwards! We've had people put the lid to the holders on upside down & then put them into the TEM. I have a GREAT service man (third party) and he phoned me from Croatia to talk me through unjamming the camera (that means it was 3 in the morning for him).
Ah, students, you gotta love 'em.
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } At 02:08 AM 1/14/98 -0500, Hermann Reese wrote: } } There is one easy way to determine if it is hardware-memory related (chip } } failure): when Windows reports the General Protection Error, one can quit } } Windows and immediately afterwards run Defrag (form root directory). Defrag } } will test the system memory on start-up and generate a warning when it } } finds faulty memory. If it finds no problems in memory, just exit Defrag } } or, even better, run it in } } full-optimization mode (can only do good to your system). } } If it does find a problem, it would indeed be best to buy new memory. } } I will have to try this test at my next opportunity, although I am somewhat } doubtful that it will catch the error. I mentioned that I had used a variety } of utilites to check memory besides the initial memory test at boot up and } the himem memory test. I used a memory check within WinProbe and a couple of } DOS utility sets. They all reported the memory to be fine. However, certain } Windows applications (like MS Word) exercised the memory in a more stringent } manner and caused failures. Perhaps the newer crop of utilities does } exercise memory in the manner that 32-bit apps do and would catch such } problems. } } And like Hermann said, running DEFRAG is a good thing anyway. Probably one } of the overlooked reasons for system slowdown. And his comments about } resource leaks is a good one. I know older versions of Microsoft Excel and } Trumpet Winsock for Win 3.x had such problems. I can pass along a shareware } app that can help you track the resources by type if it would be helpful to } anyone. } ---------------------------------------------------- } Warren E. Straszheim } 23 Town Engineering } Iowa State University } Ames IA, 50011 } Phone: 515-294-8187 FAX: 515-294-8216 } } E-Mail: wesaia-at-iastate.edu (or: wes-at-ameslab.gov) } http://www.marl.iastate.edu/marl/ (re: SEM) } http://www.public.iastate.edu/~iprt_info/cfce/ (re: coal) } } electron microscopy, x-ray analysis, image analysis, computer applications
I have had similar problems with a workstation. In my case, it related to overheating on the Pentium chip. Is your fan blowing as hard as it could?
My family and I are considering a re-location from Michigan to the Racine/Chicago/Milwaukee area. I'm looking for a general overview of organizations and/or companies which employ the use of high end microscopy such as X-ray micro-analysis and so forth for potential employment. We are curious.........any responses are highly appreciated.......
Thanks in advance
Cary Black Dow Chemical phone: (517) 636-5760 e-mail: ckblack-at-dow.com
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It's a new year, and time for the annual invitation to serve as a Guest Editor for topical papers to be published in Microscopy Research and Technique. We have finally caught up, and this Spring, will be publishing all papers within 6 month of acceptance. If you have an idea for a topical set, contact the editor-in-chief, Dr. John E. Johnson, Jr., at sdinfo-at-worldnet.att.net
While we are tossing this around, I'd like to add some comments to Steve's posting.
I certainly recognize the cost of maintaining an inventory, as well as recouping some of your engineering, manufacturing and patent (if any) costs. Your comments are valid for proprietary components. However, I had a high quote for a part which was not in stock and nor produced by the microscope manufacturer.
The component, actually considered a consummable on the service contract, failed. I called the microscope manufacturer and was quoted around $300 with a three week delivery. An internet search coughed up the location of the primary manufacturer. They quoted $30 (one tenth) with next day shipment.
You could debate if it was worth the $240 difference (we bought 2) for my half hour search and phone call, but it did save me from being down for a few weeks.
Regards,
Alan Stone
At 09:48 AM 2/17/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am in need of information concerning a replacement for 3Ms Dry Silver Recording Paper 7774, which was being handled by Ted Pella. I was told by 3M that IMATION was going to handle the paper, but IMATION says it has been suspended as a product. Does anyone know of a stockpile somewhere? I don't want to go back to wet processing! Thanks in advance for your help. Dennis
Dennis Bellotto Pathology Microscopy Services K1.210 mail code:9073 UT Southwestern Medical Center 5323 Harry Hines Blvd. Dallas, TX 75235 (214)648-3597 bellotto.dennis-at-tumora.swmed.edu "I am EM"
I have long had this tacked to my bulletin board. These are the words of the judge in a court case in which expert microscopist testimony was called upon -
"....but one general remark may be made on the microscopic testimony, and it is, that there are those who see a thing, and also those who do not see it -- those who do see it, cannot see it unless it is there, and those who cannot see it do not see it at all. But very skillful persons looking for a thing and not seeing it, creates a strong presumption that it is not there. But when other persons do find it, it goes far to displace the notion it is not there."
-- The Lord President, Torbanehill Case, 1853.
The case (involving classification of coals) was delightfully written up in an article in McCrone's Publication "The Microscope" a couple of years ago.
Dave Calvert Eastman Chemical Co. P.O. box 1972 Lincoln Street Kingsport, TN 37664 voice: (423) 229-4943 fax: (423) 229-4558 calvert-at-eastman.com
I know this topic has been discussed many times in the past, but things change rapidly, so here it is, again!
I am soliciting opinions of the Fargo Premerapro Elite at $2,000 and the Tek 450 at $8,000, as well as from fans of any other full-sized color dye-sublimation up to about $8,000-$10,000.
We already know what we are getting for ink-jet printers, so I really only need to know about dye-subs.
Mahalo for all your expertise!
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
One (or possibly two) postdoctoral fellowships are now available for the development and application of scanning transmission electron microscopy (STEM), electron energy loss spectroscopy (EELS), energy-filtered electron microscopy, and x-ray microanalysis. This biomedical research will be centered around a VG HB501 field-emission STEM equipped with EELS spectrum-imaging and EDX, and a Philips CM120 TEM equipped with an imaging filter and EDX. Both instruments are also equipped for specimen cryotransfer.
Possible research topics include: developing EELS spectrum-imaging in both the STEM and EFTEM; measuring subcellular ion distributions by x-ray spectroscopy and EELS; determining structures of macromolecular assemblies; and cryo-electron microscopy. Successful candidates should be prepared to learn and apply appropriate specimen preparation techniques (such as protein purification, rapid freezing, cryosectioning, etc).
Positions require a recent Ph.D. in biophysics or related discipline, and experience in electron microscopy. A Ph.D in physics or materials science might also be acceptable along with a commitment to apply new physical techniques to biomedicine in collaboration with other NIH laboratories.
U.S. citizenship or permanent resident status is required.
For further information contact:
Dr. Richard Leapman Biomedical Engineering & Physical Sciences Program National Institutes of Health Bldg. 13, Rm. 3N17 Bethesda, MD 20892 Tel: (301) 496-2599 FAX: (301) 496-6608 e-mail: leapman-at-helix.nih.gov
Kenneth Converse summarizes this topic quite well.
MOST service engineers are very competent, hard working, intelligent people. However there are a few bad apples in every bushel, which can spoil the image of any EM manufacturer. There are at least four methods of dealing with the service (manufacturer service contract, 3rd party service {contract or time & materials} , the Insurance company method, or service it yourself). All can work effectively and usually depend on $ and personnel.
I have personally tried each of these methods (except for the Ins. Co. method... which I researched... considered, and eventually used to bring the manufacturers to the bargaining table for service agreements which were more reasonable). My bias on the subject goes to the manufacturer service engineers, (not to slight the independent service engineers out there, cuz they're probably the most knowledgeable & versatile ones out there) but I feel the cost of servicing this equipment should be included into the sale of the instrument. I have heard this is done in some countries (Taiwan? China?), why not here in the U.S.? This would ensure proper maintenance by people who should have a vested interest in the company, it's image, and the equipment.
Currently we are not under contract with anyone, I am servicing the TEM, and when and if the scope needs a service engineer to visit/repair, we have chosen to pay time and materials. Time will tell.
Paula- I've heard of someone leaving a flashlight down the camera chamber, under the camera box, then pulling a vacuum on the whole mess. Service was required. -Mike;-)
On Tue, 17 Feb 1998, Paula Sicurello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } Dear TEM users, } } } } please take care when exchanging the TEM film magazines. Inserting the } } new magazines slightly misaligned could result in a blocking of the } } transport mechanism (like last Friday)! } } } } This usually causes more or less intensive service work! } } } } thanks } } } } Johannes } } You think that's something. We've had people put the TEM camera & the film } holder in backwards! We've had people put the lid to the holders on upside } down & then put them into the TEM. I have a GREAT service man (third } party) and he phoned me from Croatia to talk me through unjamming the } camera (that means it was 3 in the morning for him). } } Ah, students, you gotta love 'em. } } } Paula = ) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } } }
If any of you are saving quotes as they are posted, here's the citation and slight text revision of today's contribution. (Supplied by the Lawrence Hall of Science editor, who is VERY happy with your efforts.) Caroline
Hilaire Belloc, in More Beasts for Worse Children: THE MICROBE
The Microbe is so very small You cannot make him out at all, But many sanguine people hope To see him down a microscope. His jointed tongue that lies beneath A hundred curious rows of teeth; His seven tufted tails with lots Of lovely pink and purple spots, On each of which a pattern stands, Composed of forty separate bands; His eyebrows of a tender green; All these have never yet been seen But Scientists,who ought to know, Assure us they must be so ... Oh! Let us never, never doubt What nobody is sure about!
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Nothing much on Belloc, but the 1955 Bartlett's Quotations in my company library yielded up the following:
Victor Hugo (Les Miserables):
Where the telescope ends, the microscope begins. Which of the two has the grander view?
Philosophy is the microscope of thought.
Jonathan Swift:
So, naturalists observe, a flea Hath smaller fleas that on him prey; And these have smaller still to bite 'em; And so proceed _ad infinitum_. Thus every poet in his kind, Is bit by him that comes behind.
I learned this many years ago as:
Great fleas have little fleas Upon their backs to bite 'em And little fleas have lesser fleas And so ad infinitum.
It is easy to imagine that Swift had probably seen Hooke's famous drawing of an enlarged flea.
Leonard Corwin Fort Dodge Animal Health (Analytical Research) Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
Caroline Schooley wrote } And if you have "just one more", send it on!
So I sent this on to Caroline who said I should put it on the listserver.
The background is that my brother, Iain Probert, sent it to me after the X-files, a TV show, used a number of my "bug pictures" in an episode just before Christmas.
A christmas ode for the scientifically minded :
Twas the night before christmas And deep in the lab Something came crawling } From under a slab
It crawled to the agar To take a peek At all the bacteria Lying asleep
It slithered through the jelly (as icky things do) Depositing slime And gobbets of goo
Moving on to the 'scope' With a single aim To spell out a message Addressed to Elaine
'Dear Human' it wrote With some ink from a gland 'Forgot your card, Please understand'
Having studied your kind And obtained my degree On a theses titled 'Humans and their relations with me'
I find that you pry Into all that we do Without recognising We need privacy too!
You took photos of each of my chums All of my aunties And Each of my sons
You sold these photos to some t.v. show Keeping the money Isn't that so ?
If you use our photos, to give others a fright You really don't know us For it is simply not right !
Now that we've informed you Please make amends Or we'll consult our lawyers Messrs S Bends
We'll put an end to your scary sights By wearing bright colours And thick woolly tights
Viruses and bacteria Will aim to be cute And all of your ventures Will go down the chute
But heck, as it's christmas We'll give you a break Here is one scary picture And it isn't a fake !
Out of pouch, it took with a sigh, It's favourite photo Of Elaine's right eye
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
Oops!! I fowarded this message to a colleague and didn't notice that the microscopy server was still on the CC: line. I hoped you enjoyed it anyway.
} From {http://ciac.llnl.gov/ciac/CIACHoaxes.html}
Among mostly serious information it also contains the following spoof of the Good Times virus hoax that is just too good not to repost here, although it of course has nothing whatsomuchever to do with translation. Same could be said about Join the crew, I believe...
READ THIS:
Goodtimes will re-write your hard drive. Not only that, but it will scramble any disks that are even close to your computer. It will recalibrate your refrigerator's coolness setting so all your ice cream goes melty. It will demagnetize the strips on all your credit cards, screw up the tracking on your television and use subspace field harmonics to scratch any CD's you try to play....
I don't have a Tek or PrimeraPro Elite, but we do have one of the earlier PrimeraPro-s that sold fro a little more than $1000.
The short answer is, "you will get what you pay for". The Primera gave fairly nice color, but it seemed rather slow and the prints were pricey in dye-sub mode. We were running it from 486-66 machines, so things have probably improved a lot. We did not opt for the Postscript option which might have helped some. It seems the expensive dye subs may crank out the images quicker. Its fine for occasional use, but I would probably consider another option if doing a lot of color printing.
My 2-cents worth.
. At 10:02 AM 2/17/98 -1000, you wrote: } Hello, all- } } I know this topic has been discussed many times in the past, but things } change rapidly, so here it is, again! } } I am soliciting opinions of the Fargo Premerapro Elite at $2,000 and the } Tek 450 at $8,000, as well as from fans of any other full-sized color } dye-sublimation up to about $8,000-$10,000. } } We already know what we are getting for ink-jet printers, so I really only } need to know about dye-subs. } } Mahalo for all your expertise! } } Tina ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
One of my facility users has some freeze-dried sperm which she now needs to embed and section for TEM. May I ask anyone with some experience in this for advice?! I don't know specifically what she needs to see, yet.
Mucho mahalo, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
The main problem as I see it will be some kind of fixation and contrasting without rehydrating the specimens. In my checkered past we had a freeze-dryer / molecular distillation dryer / whatever that had a side access port and a small heated chamber on it. We placed small amounts of OsO4 crystals and paraformaldehyde powder in the chamber (one at a time), gently heated the chamber, opened the valve and did vapor fixation on the dried specimens. After each fixation step we re-pumped the chamber to remove as much of the excess vapors as possible.
You could probably do something similar with a bell jar or desiccator, a few feet of Tygon tubing, a test tube and a vacuum stopcock. There are a few things to watch out for:
1. Do the osmium vapor fixation first! We found that if we hit the specimens with aldehyde first it interfered with the penetration of the osmium vapors. The aldehyde probably formed some kind of "crust" or barrier at the surface of the specimens.
2. Be careful (of course) pumping out the osmium vapors. We equipped the vacuum pump inlet with a liquid nitrogen cold trap for this purpose. We also changed the pump oil very frequently. It tended to turn black quickly, indicating that the pump oil was a very good secondary osmium trap!
3. extend the infiltration time with lots of changes of resin. The osmium tended to diffuse out of the tissue and we ended up with specimens embedded in a black cloud of osmium in the final blocks. Too much of this, and it interferes with the polymerization of the resin (we used Spurr's).
Hope this helps!
All the best,
Bob ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Leica (Wild, Leitz, Bausch&Lomb, Cambridge, AO, Reichert-Jung) / Technical Instrument Company / American Volpi / Fostek / Stocker and Yale / AEI North America / OptiQuip / Dolan-Jenner / Osram / G.E. / Philips / Ushio / Boeckler Instruments / Heidenhain / Narishige / Colorado Video / Visual Environments of California, Inc. / Kinetic Systems / Pacific Precision Laboratories, Inc. / Pryor Scientific / Compumotor / Sutter Instrument Co. / Advanced Database Systems / Cohu / Javeline Electronics / Optronics / Diagnostic Instruments, Inc. / Dage MTI / Hitachi / Panasonic / Polaroid / Kodak / Mitsubishi / Sony
See if you can find a copy of Negative Staining by MA Hayat and SE Miller, McGraw Hill.
On Wed, 14 Jan 1998, Roberto Weigert wrote:
} Date: Wed, 14 Jan 1998 14:01:51 +0200 } From: Roberto Weigert {weigert-at-cmns.mnegri.it} } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM: negative staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi, } } I am interesting in the negative staining technique on whole-mount } preparation and I have some questions: } } 1) I would like to know the mechanism by which the contrasters (PTA, } Uranyl acetate etc. ) interact with the biomembranes } 2) What kind of artifacts could the contraster induce and how to avoid them ? } 3) and the same question about the putative artifacts induced by the air drying } } Thank you } } Roberto } } ______________________________________________________________________________ } } Dr. Roberto Weigert } Consorzio Mario Negri Sud } Department of Cell Biology and Oncology } Molecular Neurobiology Laboratory } Via Nazionale } S.M. Imbaro, 66030 Chieti } Italy } Phone: 0039-872-570-354 } Fax: 0039-872-578-240 } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We are looking to purchase an Image Slave system for our SEM in the near future. We would appreciate in hearing from users of this system, particularly on a Cambridge S360, on good points, bad points, advantages and disadvantages, and any other comments you may have about this system.
Look forward to hearing from you,
Rich.
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
Hi dear all, Could anyone provide me some informations about Phosphine 3R (microme n=B0 1001) used as lipid staining under fluorescence microscopy. The only information I possess is Popper (1944). Thanks to all in advance. Pascal
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 =46ax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
Hi, Our students have problems with their carbon/collodion grids; collodion to thick and/or bubling, carbon not well stticking to the plastic, not enough uniform and hydrophobic.. We are working with 2D crystals grown on lipid layers, which are transfered onto the EM grids, therefore the quality of the carbon support is critical. I've remembered that some people used to bake the carbon/collodion grids, but I do not have the reference. Any information on this method and on carbon/collodion (see other plastics) is very wellcome. TIA svetla
*----------------------------------------------------------------------* * Svetla Stoilova-McPhie, PhD | E-mail: svetla-at-bmb.leeds.ac.uk * * School of Biochemistry & | c/o EM Unit, Astbury Building * * Molecular Biology | Tel: +44 (0)113 233 3034 * * The University of Leeds | Fax: +44 (0)113 233 3167 * * LEEDS LS2 9JT | * * United Kingdom | * *----------------------------------------------------------------------*
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you might want to check out the Codonics NP1660. It does Dye sub as well = as their new "Vista" media which is black and white photo quality for 50 = cents per sheet. I have to admit I have a Codoncis 1660 but the vista = media has not arrived. The vista media requires no ribbon so it is = actually 50 cents per sheet. Of course you want to also find out when = they are shipping the vista media. They showed it at MSA in August and = it looked great. Just my thoughts, but it gives you the best of both worlds.
Judy M.
Judy Murphy, PhD Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
I know this topic has been discussed many times in the past, but things change rapidly, so here it is, again!
I am soliciting opinions of the Fargo Premerapro Elite at $2,000 and the Tek 450 at $8,000, as well as from fans of any other full-sized color dye-sublimation up to about $8,000-$10,000.
We already know what we are getting for ink-jet printers, so I really = only need to know about dye-subs.
Mahalo for all your expertise!
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************= ** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu = * * Biological Electron Microscope Facility * (808) 956-6251 = * * University of Hawaii at Manoa * = http://www.pbrc.hawaii.edu/bemf* **************************************************************************= **
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Tina, I have taken tissue that I prepared for and viewed in the SEM several times and just placed them into absolute ethanol, then embedded them normally. I have done this with very delicate specimens. Of course they were fixed and postfixed previously for SEM. If these tissue have not and you need the contrast, you probably could run the samples down a descending series of alcohols to water and then fix, etc. Another option would be to expose the sample to osmium tetroxide vapors first before going into the 100%. That maybe the only fixation you need before embedding. I would try both and compare. Good luck Hank Adams Electron Microscopy Lab New Mexico State University Las Cruces,NM 88003 phone: 505-6463600 fax: 505-6465665
I need to be able to transfer spectra acquired by my TN-5500 system to a PC, but as yet I have not be able to figure out how to accomplish it. The spectra can be stored on floppy disks but I have no means to copy the floppy disks to the PC. The PC has Kermit and FTP as a means for data and image transfer. The TN-5500 system is not connected to an ethernet card, so FTP is not viable as the means for transfer. Any help on the matter would be greatly appreciated.
List I am trying to understand FFTs of images acquired with a CCD on a TEM (what a lot of acronyms!) Can anyone recommend a book which may help me. Things like - what does astigmatism look like in the transform, defocus, periodicity etc
Many thanks
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
Speaking from the experiences of friends.... I concur with Warren. It is my understanding that to produce a relatively inexpensive dye-sub, the Fargo is mostly an electro-mechanical device, leaving most of the "computing" to your PC. If this is true, any speed evaluation of this system should use a PC similar to what you will have...
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Hello, all-
I know this topic has been discussed many times in the past, but things change rapidly, so here it is, again!
I am soliciting opinions of the Fargo Premerapro Elite at $2,000 and the Tek 450 at $8,000, as well as from fans of any other full-sized color dye-sublimation up to about $8,000-$10,000.
We already know what we are getting for ink-jet printers, so I really only need to know about dye-subs.
(1.37.109.15/16.2) id AA130053347; Wed, 18 Feb 1998 11:35:47 -0600
Responding to the message of {001501bd3c86$27597420$555b5882-at-cg.man.ac.uk} from "Chris Gilpin" {Chris.Gilpin-at-man.ac.uk} : } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } List } I am trying to understand FFTs of images acquired with a CCD on a TEM (what } a lot of acronyms!) } Can anyone recommend a book which may help me. } Things like - what does astigmatism look like in the transform, defocus, } periodicity etc } } Many thanks } Try "Transmission Electron Microscopy" by Williams and Carter, published by Plenum 1996 ISBN 0-306-45247-2 (hardbound) ISBN 0-306-45324-X (softbound)
Chapter 30 has good illustraions of FFTs of various aberrations.
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
I've just had a colleague ask me if I had any information regarding = obtaining a "reflecting goniometer". I've never used one ... but = thisperson wants to measure the angles between crystal morphologies ...
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.2106.6"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} RE: reflecting=20 goniometer {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} = {FONT=20 color=3D#000000} I've just had a colleague ask me if I had any = information=20 regarding obtaining a "reflecting goniometer". I've = never used=20 one ... but thisperson wants to measure the angles between crystal =
Gary Lovell wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I need to be able to transfer spectra acquired by my TN-5500 system to a PC, } but as yet I have not be able to figure out how to accomplish it. The } spectra can be stored on floppy disks but I have no means to copy the floppy } disks to the PC. The PC has Kermit and FTP as a means for data and image } transfer. The TN-5500 system is not connected to an ethernet card, so FTP } is not viable as the means for transfer. Any help on the matter would be } greatly appreciated.
Garry,
One way is capturing data from the printer port on the TN side with program Kermit or with my program. We have this solution for our Edax 9100 Digital LSI computer. Years ago we wrote simple program in the Basic and this program is available on the http://www.kaker.com.
-- Henrik Kaker SEM-EDS Laboratory Metal Ravne d.o.o. Koroska c. 14 2390 Ravne Slovenia Tel: +386-602-21-131 Fax: +386-602-20-436 SEM-EDS Lab http://www2.arnes.si/guest/sgszmera1/index.html MVD Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
The Electron Microscopy Laboratory of the Department of Physiology and Neurobiology at the University of Connecticut in Storrs seeks an Academic Assistant/Electron Microscopy Specialist. This laboratory provides electron microscopy services and training to faculty, staff and students. Qualifications for the position include an MS degree in a biological discipline with at least 5 years of experience in ultrathin sectioning of biological material and operation of transmission electron microscopes. Preference will be given to applicants with additional expertise in a variety of methods, such as immuno electron microscopy, analytical electron microscopy, cryo electron microscopy, negative staining and/or computer image processing. This individual must have good communication skills and be able to work independently. Duties will include: carrying out all steps in the preparation of samples, transmission electron microscopy, and photographic processing of micrographs; training of students in sample preparation and use of laboratory equipment; ordering of supplies; and routine laboratory maintenance. To apply, send resume and names, addresses and phone numbers of three professional references to: Dr. Marie Cantino, U-131, Department of Physiology and Neurobiology, University of Connecticut, Storrs, CT. 06269-2131.
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
Any reason why you can't use carbon/Formvar films? Your students may find them easier to produce using the glass slide dip method.
Regards,
Ted Dunn Hawaii
In a message dated 98-02-18 13:45:58 EST, you write:
{ { svetla-at-bmb.leeds.ac.uk To: Microscopy-at-sparc5.microscopy.com
Hi, Our students have problems with their carbon/collodion grids; collodion to thick and/or bubling, carbon not well stticking to the plastic, not enough uniform and hydrophobic.. We are working with 2D crystals grown on lipid layers, which are transfered onto the EM grids, therefore the quality of the carbon support is critical. I've remembered that some people used to bake the carbon/collodion grids, but I do not have the reference. Any information on this method and on carbon/collodion (see other plastics) is very wellcome. TIA svetla } }
{fontfamily} {param} Arial {/param} {smaller} I've just had a colleague ask me if I had any information regarding obtaining a "reflecting goniometer". I've never used one ... but thisperson wants to measure the angles between crystal morphologies ...
Neither have I. If the crystals are large enough to image in a light microscope, they can use the vernier on the circular stage which comes standard with any polarizing microscope. Alternatively, there is a small device which fits on the stage called a spindle stage which has a tiny spindle to which the crystal is attached and a goniometer to measure crystal angle. If either approach is feasible and they need further instructions, please have them contact me off-line.
I am in need of information concerning a replacement for 3Ms Dry Silver Recording Paper 7774, which was being handled by Ted Pella. I was told by 3M that IMATION was going to handle the paper, but IMATION says it has been suspended as a product. Does anyone know of a stockpile somewhere? I don't want to go back to wet processing! Thanks in advance for your help. Dennis
Dennis Bellotto Pathology Microscopy Services K1.210 mail code:9073 UT Southwestern Medical Center 5323 Harry Hines Blvd. Dallas, TX 75235 (214)648-3597 bellotto.dennis-at-tumora.swmed.edu "I am EM"
Tina, I have had the Kodak 8600 for a couple of years. I got for $6700 with ethernet, raster only when they came out with the 8650? and these were discounted!! I am truly greatful for my unexpected timing. It is an excellent printer for BW and color. So far, knock on wood, it has been a workhorse. It is networked to a PowerMac 8100/100 and parrellel-ported to a HP Vectra XU 150 . The network connection prints faster but it works great on both through Adobe PhotoShop. Good Luck with your choice. Hank Adams Electron Microscopy Lab New Mexico State University Las Cruces,NM 88003 phone: 505-6463600 fax: 505-6465665
Hi =20 Is anyone using a suitable technique which retains scales when=20 preparing SEM cross-section samples (~ 1 um scales). Is some kind of=20 coating needed to stop the scale falling off when cutting/=20 grinding/polishing ? =20 Cheers =20 f.
I have used a full reflecting goniometer. They are typically used in mineralogy (I was introduced to the technique as a student by JDH Donnay, a true guru of crystal morphology). I am sure they are, if still available, NOT cheap; you might check mineralogy supply houses. Better yet, if this is a one-time thing, find a geology/mineralogy lab with an instrument you could visit.
What you actually need depends somewhat on the crystal system; for orthogonal systems, you might get away with measuring in a microscope if you dont need high accuracy; the true goniometers have precision adjustments in 3-d space, just like X-ray goniometers, because this is necessary for studying low symmetry systems. Sad to say, we surplussed one 10-15 years ago... Good luck...
-- ******************************************************************** Pat Kingman Structural Response Team Immediate plans: Code jock....Computer hawk....Cybercadet...
It might be worth Exploring Electrolitically Coating your oxide scales with either Nickel or Copper, or preferably with both sequentially ( 1-2um Ni followed by 20-30 um Cu layer), in order to provide the necessary Support Layer to your britlle scales before, you actually attempt to slice them, grind them or polish them.
The electrolytially coated Oxide layers will then be easily handled and Examined. Let me know, if you need more specific details about the actual technique but let me know what scales are you trying to examine on what metallic sunstrates.
Is anyone using a suitable technique which retains scales when preparing SEM cross-section samples (~ 1 um scales). Is some kind of coating needed to stop the scale falling off when cutting/ grinding/polishing ?
Cheers
f. ................................................................................................. Dr. Georgios FOURLARIS Lecturer in Materials Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
} I am working with isolated Golgi membranes deposited on 300 mesh nickel } grids covered with 1% formvar (+/- carbon film), then fixed and stained } with a negative contraster. The problem is that the membranes are not } strongly attached to the film and due to the air drying they glide and } cluster togheter creating a big problem in the evaluation of the results. } } Have anyone of you the solution for my problem ? } I suggest carbon-coating the grids, then glow-discharging them for ~1 min just before putting the specimen on. Good luck. Yours, Bill Tivol
Reflecting goniometers are discussed in older books on optical crystallography. I found a treasure trove of these in the library of a nearby college with a geology department. Mineralogists are really into this kind of work.
Leonard Corwin Fort Dodge Animal Health (Analytical Research) Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
Our lab wants to do cryosectioning for TEM. We have a Reichert Ultracut E ultramicrotome. We have been told that Leica no longer makes the cryo equipment to fit the Ultracut E. Does anyone have the cryo equipment for the Ultracut E just laying around the lab? If yes, would you be interested in selling, trading or whatever for it?
Hi Pascal, I use phosphine 3R to stain neutral lipids and reference:
Clark, G. (1981) Staining Procedures 4th ed. Williams and Wilkins Co. Baltimore.
Good luck, John.
____________________________ } Hi dear all, } Could anyone provide me some informations about Phosphine 3R (microme n=B0 } 1001) used as lipid staining under fluorescence microscopy. The only } information I possess is Popper (1944). } Thanks to all in advance. } Pascal _____________________________
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D C. John Runions, Ph.D Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
Since we have been talking a lot about servicing lately...
We have a Plasmatherm Batchtop RIE in our wet lab, bought in May '96, and we want to have it serviced as it keeps coming up to atmosphere overnight. Getting someone from Plasmatherm to come up from the States will cost a lot of money, and being a small company, we don't quite have it. I was wondering if there was anyone around the Ottawa/Montreal/Toronto area who is familiar with this piece equipment and vacuum systems in general. Even someone within Canada (the poor canadian dollar makes it cheaper) would be okay.
Going one step further...if anyone near here (Ottawa) has this RIE also and would be interested in sharing costs of flying a serviceperson (is that the correct word?) that would be a big help. Forever trying to save money!
Please let me know if you have any suggestions. Thanks!
Marisa Ahmad R&D Semiconductor Insights, Inc. tel: (613) 599-6500 ext 4197 fax: (613) 599-6501
At 08:48 AM 2/19/98 +0000, frank.sarrazit-at-avestasheffield.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
At one point we developed a procedure for QC on carborized tungsten EM filaments. We found that electrolytically coating the sample for about 15 minutes really stabilized the easily-destroyed carborized coating. The result: we were able to mount, grind, and polish sample which had two phases of distinctly different hardness. An extra benefit: the copper color really highlighted the surface structure of the carborized layer in cross section.
For other types of very delicate materials, see the Buehler vacuum embedder. It is a very crafty variation on a standard dessicator, fitted with a rotatable table to carry a selection of samples and a ladle with which to pour mounting resin. Wasi Ahmed of their labs mounted a cigar with the ash still on it and was able to cross section the resulting sample without disturbing the ash!
Hope these ideas are helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite B Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis
RMC is sponsoring its Fifth Materials Ultramicrotomy Course.
The course will be September 29-October 2, 1998 in Tucson, Arizona.
This is the premier course for using microtomy for problem solving in materials sciences. The course combines a series of morning lectures with afternoon laborator= y sessions that =
bring theory and practice together. Both fundamental and advanced techniques are discussed, demonstrated, then practiced by the students.
Previous attendees have praised and endorsed the course for all materials=
scientists wanting to =
learn or refine their sample preparation skills via microtomy. =
The staff are all very experienced with problem solving in EM. The cours= e covers embedding techniques and resins, types of knives, section harvesting techniques, etching, staining, evaluating artifacts vs.real defects. The types of materials range from semiconductors to polymers, metals and ceramics.
Please call, fax or visit our web site for more information.
Steve Miller RMC 3450 S. Broadmont, Suite 100 Tucson, AZ 85713 Phone: 520-903-9366 Fax: 520-903-0132
My question relates to the focusing of a laser beam in epifluorescence confocal microscopy, i.e. when the same objective is used to both excite and collect fluorescence. The objective I'm using is 40X N.A. 1.3 oil immersion, infinity corrected.
Ideally I want to match the focus of the laser with the object plane (in-focus plane) of the objective. Clearly the convergence or divergence of the laser as it enters the back aperture of the objective will affect the location of laser focus. My current thinking is that, since the objective is infinity corrected, the laser should be parallel as it enters the objective. Does anyone know if I've reached the right conclusion about this? As a related question, is it correct to say that rays from a point source in the focal plane are parallel as they exit from an infinity corrected objective?
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I have an application requiring a considerable degree of stereological analysis (2 and 3D segments need examining):- can anybody recommend a set or sets of software that they know of or have found particularly useful/friendly for their apps.
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If the glow discharging doesn't work, you might also try treating the grids with poly-L-lysin.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Microscopists, A student in the lab is looking at DNA protein complexes on the TEM. Some references say postive staining with UA gives the best resolution, others say negative staining is fine. But the protocols all appear very similar if not the same. How is UA used to produce the two effects? And if there is any body who has direct experience with this stuff, what works best for you? How do you stain? Any opinions, suggestions, refernces are most welcome. Thank you, Kim
-------------------
Kim DeRuyter Histology and Electron Microscopy PO Box 755780 University of Alaska Fairbanks, Alaska 99775-5780 fnksd1-at-aurora.alaska.edu (907)-474-5452
Our URL is: RMC-Scientific.com/microtomes/ Our email is: RMC-at-RMC-Scientific.com =
Steve Miller, Director of Sales, North America Ultramicrotomy Products Steve.Miller-at-RMC-Scientific.com Address: 3450 S. Broadmont, Suite 100, Tucson, AZ, 85713 Phone: 520-903-9366 Fax: 520-903-0132
I look at cells in a tissue culture dish on an inverted microscope stage and would like to mark their location in the dish so I can return to that same position at a later time. I have heard of a special 'lens' that acts as a scribe that, while rotating the turret to swing it in place, it scratches the underside of the dish with a special hard tip. It has no ocular capabilities, so you cannot see as you are scratching. Does anyone know where these would be available?
Barring this cumbersome solution, has anyone solved this problem in some other way (i.e. less expensive, simpler).
judy
Judy Trogadis Eye Research Institute and University of Toronto Toronto Hospital, Western Div. 399 Bathurst St. Toronto, Canada M5T 2S8
Beth Richardson wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } hi, } does anyone have the url for RMC? } } thanks for the help, } beth } } ************************************** } Beth Richardson } EM Lab Coordinator } Botany Department } University of Georgia } Athens, GA 30602 } } Phone - (706) 542-1790 } FAX - (706) 542-1805 } Email - beth-at-dogwood.botany.uga.edu } **************************************
Address is:
RMC, Inc. 4400 S. Santa Rita Ave. Tuscon, AZ 85714 USA Tel: 602 889 7900 Fax: 602 741 2200
-- Henrik Kaker SEM-EDS Laboratory Metal Ravne d.o.o. Koroska c. 14 2390 Ravne Slovenia Tel: +386-602-21-131 Fax: +386-602-20-436 SEM-EDS Lab http://www2.arnes.si/guest/sgszmera1/index.html MVD Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
At 10:06 AM 2/19/98 +0000, brian haab wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Your assumptions are correct. In infinity corrected optics, the objective focuses on the part of the specimen which is at its front focal plane. Following basic physics, the light emerges from the back of the objective parallel (careful here!) either to the optic axis (if the feature is located on -axis) or to some principle beam which passes through the optical center of the lens (if the feature is off axis). In any event, the light will then be collected by the tube lens and imaged at the Primary Image Plane (physically located about 2 mm below the seat of the eyepiece).
Your assumptions re: the laser are also correct: The laser beam is collimated (& traveling parallel to the optic axis) on the way to the sample. It is then brought to a point of focus by the objective, at its focal plane (what would be called the "front focal plane" if looking at the information from the sample side). This diffraction limited spot is the actual "probe" for the confocal microscope.
For a more complete discussion on confocal, we strongly recommend Jim Pawley's Handbook of Biological Confocal Microscopy (2nd ed./Plenum/NY). For a more complete discussion of the difference between infinity corrected optics and fixed tube length systems, try "Optimizing Light Microscopy for Biological and Clinical Labs". For ordering info on the latter, contact me privately.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite B Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis
Beckman Institute for Advanced Science and Technology Workshop on Quantitative Approaches to the Estimation of Changes in Organs, Tissue, and Cells
Wednesday, May 6, 1998 8:30 AM-5:00 PM Beckman Institute for Advanced Science and Technology, Room 5602 University of Illinois at Urbana-Champaign
Major Speakers and Topics:
Old and New Stereology William T. Greenough, Beckman Institute
What's in a Number? New Stereological Approaches to the Quantification of Biological Structure Peter R. Mouton, Stereology Lab, John Hopkins University, Baltimore
Overview of Segmentation Techniques for Biological Staining Detection and Analysis Peter Eggleston, Amerinex Applied Imaging, Amherst, MA
Cytoarchitectonic Parcellation of Cerebral Cortex Joseph Nunez, Department of Psychology, University of Illinois
Stereology I: Point Counting and Cycloid Grid Anna Klintsova, Beckman Institute
Stereology II: Optical and Physical Disector Janice Juraska, Department of Psychology, University of Illinois
Densitometry: Concepts and Pitfalls Scott Irwin, Neuroscience Program, University of Illinois
Densitometric Evaluation of Immunocytochemical Reactions and in situ Hybridization Roberto Galvez, Neuroscience Program, University of Illinois
Image Analysis Software Bridget Carragher, Beckman Institute
There will also be time set aside for a hands-on demonstration of stereology, with Nick Kisseberth, Joseph Nunez, Anna Klintsova, Beckman Institute; and for a demonstration of semi-quantitative densitometry analysis of radiolabelled in situ hybridization reaction, with Marc Cohen, Neuroscience Program, University of Illinois.
Workshop Organizer:
Anna Klintsova, Research Scientist, Beckman Institute
Registration Information:
The workshop is limited to 65 participants, taken on a first-come, first-served basis. A $30.00 workshop fee covers the costs of continental breakfast, lunch, coffee breaks, and materials. Participants are responsible for their own travel and lodging. Checks should be made out to the "University of Illinois."
There are ten (10) student fellowships available. The workshop fee will be waived for those receiving a fellowship. To apply for a fellowship, the student must write a brief statement about why the workshop is important to his/her research.
Register by sending your name, address, telephone number, fax number, and email address, and whether or not you will need parking; as well as your fellowship statement, if you are applying for a fellowship, and your check to:
Julie Weaver Beckman Institute University of Illinois 405 North Mathews Avenue Urbana, IL 61801 Tel.: 217-244-4906; Fax: 217-244-8371 Email: jweaver-at-uiuc.edu
DEADLINE for both registration and fellowship application is April 15, 1998. Your registration will be considered complete when we receive your check. Registration and fellowship acceptance will be confirmed by April 22.
I'm posting this request for a friend of mine. He's looking for the schematics and any technical information (diagrams, etc.) for a Pfeiffer pumping unit control TCS100 for a Balzers BAF300 or BAF301 freeze etch unit (it's a high vac unit).
I guess he's trying to rebuild it & it's kinda old & of course the info. did not follow the machine.
Thanks for all your help.
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
To whom it may concern, I am currently in the market for a Hitachi 600AB or a Jeol 1200CX TEM.This instrument will primarily be used for asbestos analysis.
Thank you,
Kevin MacKinnon Micro Analytical Laboratories, Inc. Emeryville, CA 94608 (415)653-0824
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Here is my experience on the inside of the pricing of spare parts for service. I have worked for several instrument companies over the years in service support engineering and have been on the front lines of the "how much to charge for parts" problem many times. Nominally spare parts prices were set so we could could make a fair margin to cover costs and profit while maintaining a fair price for customers. As a service advocate I wanted low prices to improve customer satisfaction, but for the company the real object of the game was to maximize the selling price without making the customers go away.
The problem starts because most companys' CFOs want the cost of goods to be about 25% to 33% of the selling price. In most instrument manufacturing businesses this ratio is observed for the finished produts. They also extend this policy down to spare parts. Thus if you doing the pricing of small spare parts you are usually are told to make the price around 3X the cost. As the cost of the items goes up the multiplier will go down but there is still a big markup.
For some items I would find that a spare part would go through 2 or three markups before we sold it as a replacement part. For instance a small part might be priced for $30 by the orginal manufacture and sold to a distributor. That distributor marks it up to $90 for sale to the instrument manufacturer as a spare part. It gets marked up again by 3X and becomes a $270 part!
The practice of multiple markups is worse with Japanese companies. In many cases, orders for spare parts may go through serveral companies and mark ups before thay reach the USA service organization from the factory. And that is also why it takes so long to get parts from Japan!
I know this sounds awful, but it is a real practice at many of our instrument companies. It does pay to find the original manufacturer for these parts.
Disclaimer: Now that I run my own business I don't do it that way!
Ronald Vane XEI Scientific
alan stone wrote: }
} While we are tossing this around, I'd like to add some comments to Steve's } posting. } } I certainly recognize the cost of maintaining an inventory, as well as } recouping some of your engineering, manufacturing and patent (if any) costs. } Your comments are valid for proprietary components. However, I had a high } quote for a part which was not in stock and nor produced by the microscope } manufacturer. } } The component, actually considered a consummable on the service contract, } failed. I called the microscope manufacturer and was quoted around $300 } with a three week delivery. An internet search coughed up the location of } the primary manufacturer. They quoted $30 (one tenth) with next day shipment. } } You could debate if it was worth the $240 difference (we bought 2) for my } half hour search and phone call, but it did save me from being down for a } few weeks. } } Regards, } } Alan Stone }
Hi The lab.of Electron Microscopy, Faculty of Bioengineering, National University of Entre Rios, Argentina, is searching an AFM to received it in donation. The microscope will be used in research & development and teaching. We pay all the charges of the shipment. Please, contact to
=================================================== Fernando D. Balducci Laboratorio de Microscopia Electronica Facultad de Ingenieria - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
McCrone Accessories & Components, 800-622-8122, has a device that marks a ring with ink on the glass. It is not clear from the catalog whether you can see through it. It mounts with RMS threads.
Leonard Corwin Fort Dodge Animal Health (Analytical Research) Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
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I look at cells in a tissue culture dish on an inverted microscope stage and would like to mark their location in the dish so I can return to that same position at a later time. I have heard of a special 'lens' that acts as a scribe that, while rotating the turret to swing it in place, it scratches the underside of the dish with a special hard tip. It has no ocular capabilities, so you cannot see as you are scratching. Does anyone know where these would be available?
Barring this cumbersome solution, has anyone solved this problem in some other way (i.e. less expensive, simpler).
judy
Judy Trogadis Eye Research Institute and University of Toronto Toronto Hospital, Western Div. 399 Bathurst St. Toronto, Canada M5T 2S8
I have to agree with Owen here. I have run EMs both under manufacturer service contract and self-serviced by an in-house engineer. I have seen excellent, cost-effect service from some companies, and execrable call-them-when-you-want-it-broken service from another company. So a thing to note: if you go 3rd party, from where do they hire their engineers? If the engineers are mostly hired from manufacturers with whom you've had poor service experience, then you're not likely to be happy with the 3rd party, either. If the 3rd party engineers come from a manufacturer with good service, then you likely will be happy. Barring company administrators, of course.
Also keep in mind that service can vary with geography, because personnel does. Not just engineers, but administrators. Recall that bad administrators can break the best engineers.
One thing I haven't seen mentioned: at one time, a 3rd party service company (whom I won't mention, as this was 2-3 years ago, and they may not be doing this anymore) offered me a service contract with a rebate for the "insurance" part of the contract. That is, if we didn't need any parts, most or all of that part of the contract was either rebated or applied to the next year's contract. This should be more common for both 3rd party and manufacturer contracts.
My conclusions are that service is "...a MAJOR consideration in the decision to purchase a [brand of] instrument...". But also, the cost-effectiveness of in-house service should be part of the decision. People have discussed the relative costs of manufacturer vs 3rd party service (or insurance), but the costs of downtime and do-it-yourself must also be considered.
In all the discussions of service contracts, I don't recall ever seeing an actual cost-accounting of the various service options. This would include not only downtime and employee costs for in-house work, but the cost of operating a scope at less-than-spec because the service isn't good enough to return the scope to operating conditions, the costs of getting the scope tweaked to the best possible performance when there is no actual requirement for ultimate resolution, the cost of replacing an entire X thousand dollar circuit board instead of the $20 IC chip that went bad vs the hours spent a) finding the bad chip, b) finding a place to buy 1 or 2 chips instead of the 100s or 1000s that manufacturers buy, c) verifying that was the problem, and d) repeating the cycle to fix the problems [often on the same board] uncovered by replacing the first part. And many more such considerations.
I particularly agree with Owen's last comment about the lack of respect for service engineers (except for one company).
Phil disclaimer: since I'm an editor, I'd love to have someone write an article for us about how to do cost-accounting as part of a purchase decision.
} My two cents. } } Service should be a MAJOR consideration in the decision to purchase an } instrument. The extra bells and whistles that swayed you towards a } particular brand aren't worth much later when your scope is down for other } reasons. } } Also, this thread began with a survey of service contract options. One } overlooked option is doing some of the service yourself and taking } advantage of discount service contracts offered by most manufacturers. By } doing some of your own repairs, you may 1) save some money, 2) learn more } about your instrument, 3) develop some empathy for field service engineers } (who aren't getting much respect in this discussion). } } Owen } } Disclaimer: My employer hired me because of the views expressed in this } message. } } Owen P. Mills } Michigan Technological University } Metallurgical & Materials Engineering } opmills-at-mtu.edu
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 5037 Station A Champaign, IL 61825-5037 oshel-at-shout.net or poshel-at-hotmail.com
At 04:52 PM 2/19/98 -0500, Judy Trogadis wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When I used to work at Cambridge Instruments (now Leica), they sold a marking objective which you could rotate into position. It just had a simple ink stamp on it. I only had my hands on it once but as I remember, it had a collar on it and you just moved the marking ring toward the slide/dish and gently pressed to leave an impression. I'd suggest you try your local Leica rep or dealer.
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite B Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis
Hello We want to scan series sections from myocardial tissue directly from the histological slide into our computer. What scanning resolution (dpi) do we need in order to recognize a fiber with a diameter of about 25 =B5m? Thank you Dr. Klaus Redmann Experimental Heart Surgery University Clinic M=FCnster, Germany redmann-at-uni-muenster.de
Folks: I believe Michael Shaffer is referring to a four axis stage for a light microscope. This stage is mounted on a mic. used for Petrographic examination of geological thin sections. We have such a stage for a Zeiss microscope. It has the stage, several glass spheres for light diffraction and a low ma condenser. This stage cost over $7k US when it was purchased. We no longer have any use for the stage and would like to either sell, trade, barter it. Our preference would be to see it go to a research/ teaching group. We are willing to entertain any inquiries. Regards; Jon McGovern J. P. McGovern and Associates e-mail: jmcgover-at-cadvision.com
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RE} TEM wanted 2/20/98 8:41 AM Dear Kevin,
The McCrone Research Institute in Chicago is selling their Jeol 1200EX. I= t is used primarily for asbestos analysis. Anyone interested should conta= ct John Shane (312-842-7100) or see the website at www.mcri.org. I will b= e posting the ad soon.
thanks.
John Shane Director of Research McCrone Research Institute
--------------------------------------
To whom it may concern,=20 =09I am currently in the market for a Hitachi 600AB or a Jeol 1200CX TEM.This instrument will primarily be used for asbestos analysis.=20
Thank you,=20
Kevin MacKinnon Micro Analytical Laboratories, Inc.=20 Emeryville, CA 94608 (415)653-0824
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
id sma000173; Fri Feb 20 16:30:41 1998 Received: from c1pa008.lkasbg.gv.at (c1pa008.lkasbg.gv.at [10.1.42.8]) by hermes.lkasbg.gv.at (8.8.5/8.8.5) with SMTP id RAA21071; Fri, 20 Feb 1998 17:32:33 +0100 Received: by c1pa008.lkasbg.gv.at with Microsoft Mail id {01BD3E1D.59359760-at-c1pa008.lkasbg.gv.at} ; Fri, 20 Feb 1998 16:34:00 +-100 Message-ID: {01BD3E1D.59359760-at-c1pa008.lkasbg.gv.at} Pascal Veys {veys-at-bota.ucl.ac.be}
Salzburg, 20th Febr. 1998, local time: 3.55 p.m.
Dear Pascal, hope, some of the information given below can be of help:
1) ex: BANCROFT J.D., STEVENS A. (Eds), Theory and Practice of = Histological Techniques, 4th Ed., CHURCHILL-LIVINGSTONE, 1996; ISBN = 0-443-04760-X: only subject index reference to "phosphine-solubility = (=3D appendix VI, Solubility of some Dyes; p.739): } } Phosphine: generic name: Basic Orange 15, 3rd ed. CINo.: 46045, = soluble in water; soluble in alcohol. { { {
2) ROMEIS, Mikroskopische Technik, 17ed, 1989 (URBAN&SCHWARZENBERG = Publ., German): p. 377: (translated):=20 subchapter: Fluorescence Microscopy " Only lipopigments like Lipofuscin, Carotinoids and Vitamin A show = Auto- fluorescence. Secondary fluorescence only can be obtained by vital = or subvital staining with fluorescent dyes. Cell Cultures, = tease-preparations, as well as unfixed cryostat-sections are/can be = treated for 3-5 minutes with/in a 0.5% aqueous solution of } Rhodamine B { = or } Phosphine 3R (Popper, 1951, see below); after staining washing in = isotonic salt solution (NaCl, physiological), mounting in = glycerol....Lipids, cholesterol-esters included, will display a = silvery-white to green fluorescence. Fatty acids, soaps as well as = cholesterol will not be evidenced. POPPER 1951 (see above) in the reference list of this book is cited only = as=20 POPPER H. (1941): Histologic distribution of vitamin A in human organ = under normal and pathological conditions. Arch. Path. 31, 766-802.
3) Old, but a source of "treasures" for stainings: LILLIE RD, FULLMER HM = (Eds) 4th Ed. Histopathologic Technic and Practical Histochemistry, McGRAW HILL 1976 p. 144/145:=20 subchapter Fat Stains: } } In fluorescence microscopy several acid and basic dyes have been used = in simple aqueous solution of fat stains. Of these, PHOSPHINE, a basic = acridine dyestuff seems to be one of the best. Also used are..... { { = Informations contained from subchapter "Staining of Lipids with esterified Sudan dyes" (p. 571, = 572) FLUORESCENCE MICROSCOPY: .......POPPER (1941) used either..... or a 3 = min stain in 0.1% aqueous Phosphine (CI 46 0 45).......produced a = silvery white fluorescence and demonstrated more fats than traditional = strong alcoholic Sudan methods..... { { {
Maybe I could go on giving more informations, but I shall stop here; if you want to get more informations (chemical structures, etc., more = procedures, etc....) you are kindly invited to specify more detailed = your needs on the dye....
Hope this helps for the moment, best regards
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
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Hi Pascal, I use phosphine 3R to stain neutral lipids and reference:
Clark, G. (1981) Staining Procedures 4th ed. Williams and Wilkins Co. Baltimore.
Good luck, John.
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D C. John Runions, Ph.D Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
At 04:38 PM 2/19/98 -0800, Paula Sicurello wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have that. How do you want to get it?
Rick A. Harris, Director Microscopy and Image Analysis Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 752 3085 fax raharris-at-ucdavis.edu
} 3M corporation has quit making Dry Silver Recording Paper #7774. A } friend of mine uses it to print his electron micrograph negatives and } is in desperate need of this type of developing paper. We have talked } to everyone under the sun, cruised the Internet and Web sites without } any luck. Kodak does not make it. Does anyone know of a supplier or } does anyone have a hugh stash they might be willing to sell, trade, } barter, etc?
This paper was discontinued because it contains mercury and the prints were considered toxic waste and have to be treated appropriately.
John Bozzola 71 Concordia Drive Carbondale, IL 62901 Internet address (SIU): bozzola-at-siu.edu Internet address (home): JBozzola-at-aol.com
Since my last posting about transfering TN-5500 spectra to a PC I have managed to send spectra information( headers and each channels intensity) as ASCI data to the PC. I have not however found a means to convert the data into a graphics file. Kermit was used to capture the xfered data. The PC is connected to a Novel network but the only means of TN-5500 data or spectra to be xfered is by means of a serial port connection beween the two systems. Any information concerning software that can be located on the PC for converting the data to a graphics file and then converting the graphics file to a TIF file would be greatly appreciated. If anyone is interested, the spectra data was xfered by using Noran's XI module(file type 5, number 442), which requires two other type 6 files( 4000 and 4065 ) be present on the operating system.
You may be doing about as well as you can. Those old PDP-11s can be setup on the ethernet for a price in order to use FTP to ship files under the TCP/IP protocol. But I don't know of any PDPs being set up to do Novell. Kermit may be the mose expedient solution.
When I did similar transfers from our Kevex, I imported the data into Excel for graphing. I had to parse it properly which isn't too bad if the channel counts are one per line. You could then "fancy up" the chart with whatever format and labeling you like. The later versions of Excel also allow you to define customized chart types and to set the default chart type for new charts. Once you have that set up, it should be quite straightforward to convert the data over.
Since the chart would be prepared in Excel (or something like it), you would not have to save it as a bitmap (TIF or BMP) unless you really had to. It would be better to keep it in chart format or at least as a picture or windows metafile format in order to retain the fine detail and allow for possible editting later.
Then again, maybe somebody has more details of a customized program to do all this in one step.
At 11:05 AM 2/20/98 -0600, you wrote:
} Since my last posting about transfering TN-5500 spectra to a PC I have } managed to send spectra information( headers and each channels intensity) as } ASCI data to the PC. I have not however found a means to convert the data } into a graphics file. Kermit was used to capture the xfered data. The PC } is connected to a Novel network but the only means of TN-5500 data or } spectra to be xfered is by means of a serial port connection beween the two } systems. Any information concerning software that can be located on the PC } for converting the data to a graphics file and then converting the graphics } file to a TIF file would be greatly appreciated. If anyone is interested, } the spectra data was xfered by using Noran's XI module(file type 5, number } 442), which requires two other type 6 files( 4000 and 4065 ) be present on } the operating system. } } ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
The New England Society for Microscopy announces its February Meeting
Dear fellow microscopists,
At 09:16 AM 2/20/98 -0400, Leonard Corwin wrote:
} McCrone Accessories & Components, 800-622-8122, has a device that } marks a ring with ink on the glass. It is not clear from the catalog } whether you can see through it. It mounts with RMS threads.
I didn't respond to this on the first go-round, because I didn't think that it was what the orinal poster (Judy) wanted, but we (Energy Beam Sciences) have made a device like this, called the Slide Marker, for over 20 years.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
At 4:52 PM 2/19/98 -0500, Judy Trogadis wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Judy
Ernest F. Fullam, Inc sells a device similar to the one you describe. The the toll free phone # is 800-833-4024 or you can try 518-785-5533. Fax# is 518-785-8647. The item is called an EFFA microdotter, and the catalog ('92--'93) # is 13400. Good luck1
I think a point of clarification is needed here. The instrument described by Michael Shaffer is an optical goniometer. It is used to measure the angles between the faces of crystals. The crystals, usually small (i.e. mm sized) are mounted on a graduated rotating stage and illuminated through a source that is coaxial with an observing telescope. By rotating the stage so that first one, then another face is illuminated (i.e. reflecting), and by taking readings on the stage, it is possible to obtain interfacial angles to {1 degree. These devices were once in common use among crystallographers, but I have neither seen nor used one for 20 years. Perhaps the best chance Michael Shaffer has to get one is by asking at the nearest geology department, they may have one (gathering dust?) somewhere.
The instrument described by Jon McGovern sounds like a universal stage. This is an attachment (for a polarizing microscope) that is used to rotate a thin section (of a transparent or translucent optically anisotropic solid) so that its optical directions can be located. The glass hemispheres are used to "sandwich" the section during observation. Normally objectives with a long working distance will be needed because the glass hemispheres tend to be quite large.
Hope this helps.
Roger Mason
At 08:34 AM 20/02/98 -0700, Jon McGovern wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
At 09:09 AM 2/19/98 -0500, Pat Kingman wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One more note on this topic: In microscopy terms, we frequently refer to this type of accessory as a "universal rotating stage". It is useful in both orthoscopic and conoscopic observations (i. e., maximum birefringence, crystal angles, etc.).
As I understand it, the glass hemispheres come in different refractive indices, chosen to correlate with the refractive indices of the mineral/material under study. From the descriptions, it also appears that you need ultralong working distance objectives and condensers. And, if you are going to do conoscopic work, the NAs need to be large.
I quickly found two good references in our library ( I am sure there are more): 1. A treasured (but out of print) booklet from the old Leitz: "Polarized-light Microscopy: Principles, Instruments, Applications" by Walter Patzelt (p. 87) 2. Hartshorne, N. H., and Stuart, A. Crystals and the Polarising Microscope, 4th ed., Arnold (London), 62-63. I noted with interest that Hartshorne and Stuart mention a simple instrument, designed by Wollaston in 1809 (!) that strongly resembles the spindle stage designed by Don Bloss (available, I believe, through McCrone Associates).
In any event, good luck with the hunt!
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite B Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis
Does anyone on this listserve know of recent salary surveys for microscopists (EM, etc) in the university and private/ industrial sectors? The last one of know of was published by EMSA in 1984. Anything more recent would be helpful. TIA Hank Adams Electron Microscopy Lab New Mexico State University Las Cruces,NM 88003 phone: 505-6463600 fax: 505-6465665
We specialize in just SEM. Our goal is to provide high quality results without high cost. We use digital imaging for flexibility and to decrease turnaround time. Visit us at http://www.freeyellow.com/members/smartfelten/ If you are an overbooked SEM operator, we would love to help you out! ========================================================================== == === Ric Felten Owner SMARTech PO Box 439 Goshen CT, 06756 (860)491-3299 e-mail: rfelten-at-esslink.com
A couple of timely news releases today shed some further light on the question of the reliability of Zip drives, from our earlier thread on this subject.
In the first, Paul Festa of CNET NEWS.com reported on the official report from Iomega that only a fraction of 1 percent of Zip drives have exhibited any problems. The article is at:
http://www.news.com/News/Item/0,4,19342,00.html
In the second, Dan Briody of InfoWorld Electric wrote an article titled "Goodbye floppy? Micron offers Zip drive standard." It reports the March shipment of Micron's Millenia line of PCs which will offer Zip drives standard as the a:drive, with 1.4MB floppies as optional. The article is at:
It seems to me evident that a major computer manufacturer with a long time experience in installing Zips in their computers would probably not make the jump to offer Zips as a standard replacement for the floppy if there were any *real* problem with the reliability of the drive.
I am happy to hear that the failure rate is below 1 percent. I figure that there are 20-25 Zips presently in use in my group, so we can buy another 75 drives before we should expect one of them to fail....
Larry
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
There is one more factor besides reliability in replacement of floppy drives with zips.
Trouble shooting and computer maintenance....
You can boot your Hard drive from a floppy, but unless something is drastically changed , even using Norton Utilities 3.0 you can not boot up off a zip. Though Norton Utilites 3.0 lets you reboot a Win 95 system from a zip AND a floppy.
Something to watch for when wanting to toss your floppy drive.
..... Speaking as one who has had to do Hard Drive crash recovers...
Lou Ann
} It seems to me evident that a major computer manufacturer with a long time } experience in installing Zips in their computers would probably not make } the jump to offer Zips as a standard replacement for the floppy if there } were any *real* problem with the reliability of the drive. }
*************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
Microscopy Home Page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/CSMS/csms.html
Personal Home Page: http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/LAM.html
Dear Listers, I'm seeking input from those who are currently using either RMC's MT-XL/CR-X or Leica's UCT-FCS cryoultramicrotome. It would be housed in a multi-user facility and primarily used for biological samples (plant, animal including insects)polymers and fibers. The information I seek is: a) reliability of instrument b) maintenance / service during and after warranty
TIA Rosemary
#################################################### Rosemary Walsh Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:rw9-at-psu.edu ####################################################
I don't know about PCs, but this is not true for Macs. Put a system folder on a Zip disk, either by Installing one or copying one over and "blessing" it--open, then close the folder. When starting the Mac, hold down command+option+shift+delete (DOCS), and the Mac will start up from the Zip drive.
Macs will happily start up from their internal hard drive, an external hard drive, a CD-ROM, a floppy, a Zip, Jaz, Sy Quest, paper towel, or anything else as long as the device is readable by a Mac and has a blessed system folder on it. Norton or any other utility program not needed, this is a feature of the Mac OS.
Speaking as some one who has done this [well, not the paper towel ... 8-) ] .
Phil
} There is one more factor besides reliability in replacement of floppy } drives with zips. } } Trouble shooting and computer maintenance.... } } You can boot your Hard drive from a floppy, but unless something is } drastically changed , even using Norton Utilities 3.0 you can not boot up } off a zip. Though Norton Utilites 3.0 lets you reboot a Win 95 system from } a zip AND a floppy. } } Something to watch for when wanting to toss your floppy drive. } } ..... Speaking as one who has had to do Hard Drive crash recovers... } } Lou Ann } } } It seems to me evident that a major computer manufacturer with a long time } } experience in installing Zips in their computers would probably not make } } the jump to offer Zips as a standard replacement for the floppy if there } } were any *real* problem with the reliability of the drive. } }
I replied to Lou Ann Miller's message this morning, but forgot to cc the listserver. Here is again:
Lou Ann:
I'm afraid you are slightly behind times on your information about booting off a Zip. First, you have *always* been able to boot off a Zip (or other peripheral drive) if you used a Mac (as I reported earlier, one of my colleagues used a Zip exclusively for quite a long time when her hard drive crashed). This is now the case on PCs also...again, I am sure that Micron Electronics would not be putting Zips as the a:drive on an entire line of their product, and making floppies optional, if you could not boot from the Zip....
Larry (So, is *this* the last word?.... ;-) )
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Recently returning to the EM fold after an absence of several years, I have been unhappy with the staining of LM sections with Toluidine Blue/Borax or Tolouidine Blue/Basic Fuschin.Buried in a cupboard however I found a bottle labelled Richardsons Stain which seems to work very well. However I have no idea whats in it, (its either Toluidine Blue or Methylene Blue based).I am loathe and embarressed to admit that the bottle is labelled in my own handwriting, however advancing years and alzheimers have combined to defeat my memory. Any help would be much appreciated.
Peter Smith AgResearch Wallaceville Upper Hutt New Zealand
see: RICHARDSON KC., JARETT L., and FINKE EH: Embedding in epoxy resins for ultrathin sectioning in electron = microscopy. Stain Technol. vol } 35 {(1960), 313-323 Maybe this receipt would fit your needs.
But: there is another staining formula, know to me for a long time (I = used it as a student in the 1970ies) also as "Richardson's staining", = essentially a polychromatic staining for Epon resin sections, containing = Methylene blue and Azure II. The formula I got from a technician at the = University of Salzburg, at that time the formula was written into the = lab handbook there. I searched for the origin of that formula /method = and the only facts I got were: "after MALLORY (& RICHARDSON KC et al. 1968)" but I was not able to = verify any paper dealing with such a staining procedure as follows:
Essentially it is an=20 aqueous Methylene blue - Azure II in Borax-buffer (high pH, appr. = 9-10). Solution prepared preferably in an Erlenmeyer-flask
1) prepare 2% Borax (di-sodium-tetraborate: Na2B4O7) in a.bidest. 2) after complete dissolution of Borax-powder: add 1 g methyleneblue, add 1 g Azure-II-dye (would recommend to use certified dyestuff) 3) mix well/gently agitate (magnetic stirrer) let stand and/or mix at = least over night (to get saturation of the dyes as complete as possible) 4) filtrate the solution into a tightly (pref. screw capped) closeable, = brown lab-bottle (100-250 ml, light protection necessary): use as = working solution, but filtrate solution before staining sections (either = by adding drops of staining solution to the slide/section, then heating = on the hot plate to the boiling point: but don't let cook!, jet washing = with squeeze-bottle) The staining solution ready for use is stable for several months (even a = year).
Note: I personally used this stain successfully for staining 2 ?m = semithin Epon sections of whole rat hypothalamus for a long time.=20 But I had to change to another polychromatic staining (which I apply = very successfully now for 6 years and as another, earlier modification = another 7 years before) due to problems in stainability of = "Epon"-sections when the original "EPON 812 R[superscript] = (SHELL/MONSANTO)" was not available any more (suspected changed = polymerization properties...chemical nature.....).
Therefore it would be interesting to hear of your special = problems/antipathies in staining sections with Toluidineblue/borax or = toluidineblue/basic fuchsin (which I assume to be diluted in alkaline = buffer solution too). Also it would be of utmost interest which type of = resin (name, dealing company) you currently are using.
NOTE 2: Hildegard H. CROWLEY, also described a "SUPERIOR METHYLENE BLUE-AZURE II Stain for semithin sections" which essentially is the same mixture as above, and Leslie WESTON also = had a posting for that as follows:
MSA-Listserver 14th Oct. 1997: ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Are we talking about Richardson's stain? (I don't have the reference to hand, but I could find it if anyone wants.) This is a 1:1 mixture of 1% methylene blue in 1% borax with 1% Azure II in water. It is my routine stain for thick sections and gives brilliant metachromatic results, even without an alcohol rinse.
Lesley Weston (e-mail: lesley-at-unixg.ubc.ca )
On Tue, 14 Oct 1997, HILDEGARD CROWLEY=20 e-mail: hcrowley-at-du.edu
wrote:
} = ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of = America=20 } To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com } = -----------------------------------------------------------------------. } =20 } =20 } Hi, } =20 } The CI number for Methylene blue is 52015. } The CI number for Azure B (sometimes called Azure I) is 52010. } There is no CI number for Azure II since it is a 1:1 mixture of the = the=20 } two stains above. } Could you simply mix 52015 and 52010? I have not done this, simply=20 } because the Azure II worked fine, and I don't have time to fix things=20 } which already work reliably. I cannot think of any reason why mixing = the two=20 } stains rather than buying the azure II would alter the situation. } We buy our methylene blue and our Azure II from Sigma. I do not know = if=20 } the percentage of active stain varies per gram between batches. This = is=20 } something one=20 } needs to be aware of given the vast interactions between LM stains and =
} embedding epoxies. I also used to buy very nice stains from Fisher = (but=20 } someone stole our Fisher Catalog)! PS. I do not have any commercial=20 } interest in either of the companies mentioned. The price of these = stains=20 } can vary enormously so it pays to check several sources. Just make = sure=20 } that the company states the CI number of the product they are selling. } Please note that the stain improves with age. Make a lot and let it = sit=20 } in the dark at rt. Also please note that if the stained section is = not=20 } rinsed with alcohol (75% - destained, and then with 100%), the result=20 } will not be as brilliant as it should be. =20 } Bye, } Hildy } =20
Hope this helps, if any questions more: require
Best regards and have a good day Wolfgang
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
Recently returning to the EM fold after an absence of several years, I = have been unhappy with the staining of LM sections with Toluidine = Blue/Borax or Tolouidine Blue/Basic Fuschin. Buried in a cupboard = however I found a bottle labelled Richardsons Stain which seems to work = very well. However I have no idea whats in it, (its either Toluidine = Blue or Methylene Blue based). I am loathe and embarressed to admit that = the bottle is labelled in my own handwriting, however advancing years = and alzheimers have combined to defeat my memory. Any help would be much = appreciated.
Peter Smith AgResearch Wallaceville Upper Hutt New Zealand
This has been a remarkable thread, full of input from a variety of sources. I think that we have managed to show that there are a variety of possible solutions to the problem, but first, let's consider the options.
If the responses are indicative, and my own experience shows that they are, service from any source is spotty at best. It basically comes down to the service engineer that you will commonly deal with. An SEM is one of the most complex instruments manufactured. It requires a unique individual to not only provide for appropriate service of an SEM but to also provide a customer orientation that can match itself to the expectations of a user. I can confirm that there are those who work for manufacturers who can maintain this balance, as there are those who work for themselves or third party services.
Ken Converse mentioned some of the requirements and costs of maintaining a service engineer, but he was vague in his descriptions. I have seen companies that are very comfortable in justifying spending upwards of $250,000 a year for service engineers, in salary, benefits and travel expenses. Needless to say, the engineer's salary and benefits are the lessor in this equation. At the same time, these same companies will extend additional thousands or tens of thousands of dollars for bringing new hires to the factory for 'factory' training, even if the facility is out of country.
This points to a very inefficient method of training and resource management. At the expenditure levels of some companies, it would make far more sense to provide 4 times the number of service personnel and back those field engineers with a secondary level of expertise of technical specialists and field trainers. I have actually witnessed service organizations whose secondary level of expertise required importing personnel from overseas. A very inefficient use of manpower and a drastic increase in the departmental expenses.
On the other hand, third party services are often taking advantage of the high service costs of the manufacturers. Given the markups, it is very easy to underbid the manufacturers by 10% - 20% and still make a very good living. By undertaking third party service, though, you are opening yourself up to the experience and knowledge of a limited group of field engineers. While the basic engineering of EMs has not changed for years, there can be significant differences in the designs of individual manufacturers. Finding service engineers who can understand and adapt to these differences can be difficult.
The 'HMO' type insurance agencies can, perhaps, offer even greater savings. However, they come with even greater risks. There is no guarantee who will be servicing your instrument. These agencies will pit the manufacturers and the third party sources against each other, and award service to the party that offers the lowest price. There will be no guarantee that the same service personnel, or even the same company, will provide service from call to call.
It basically comes down to how much responsibility you are willing to take for the maintenance of your instrument. There are those who have posted here who do most of the primary service themselves. I basically promote this approach. The more you know about your instrument, the better you can make decisions on the service of it. Being directly tied to the maintenance of your instrument makes you far more informed on the service options available to you.
It is all too common, though, for users to take the easy course of relying on the manufacturer for service. Just as it is all too easy for those management types out there to go for the 'HMO' type service management organizations who offer to cut service costs by attractive amounts. I firmly believe that third party service organizations can offer a reasonable middle ground - consistent service personnel along with flexible contract and billable terms. In most cases, you will be married to your instrument for some time. Try the various alternatives and make up your own mind as to the suitability to your needs. However, think ahead and protect yourself in two ways - demand complete documentation when purchasing - including full schematics, and demand service guarantees when shopping around.
Once again, I am speaking from a biased viewpoint - I am a third party service provider for SEMs and other equipment. However, this being such an esoteric field, I believe that it is especially important that all sources be exploited and supported since the competition can only benefit everyone.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
I found the Richardsons Stain in P. Boecks book "Der Semid=FCnnschnitt":
1% Azur II in A.dest 1% Methyleneblue in 1% Borax mixed 1+1
Dry sections, staine for 1-2 minutes at 60 degrees (hetating plate), rinse slides in A. dest, drying and mounting (immersion oil or whatever).
The original method is described in: Richardson KC, Jarett L, Finke EH (1960) Embedding in epoxy resins for ultrathin sectioning in electron microscopy. Stain Technol. 35: 313-325
Probably there are some modifications of this method, but I hope this helps.
Jens
--------------------------------------------------------------- Dr. Jens Buecking Tel. +49-(0)421-218 3745 University of Bremen Fax. +49-(0)421-218 4620 Dep. of Biology Email jbueck-at-biologie.uni-bremen.de Leobener Str. - NW2 28359 Bremen ---------------------------------------------------------------
INTERNATIONAL COURSES OF LIGHT MICROSCOPY, PHOTOMICROGRAPHY AND LASER SCANNING CONFOCAL MICROSCOPY GARGNANO (Lake of Garda) October 1998
The Course is a post-graduated theoretical/practical course, with propedeutical lectures and practical stages on microscopy, photomicrography and confocal microscopy. The course will take place in Gargnano (Lake of Garda) in October 1998.
All information and registration details (participation fee, date, special accomodation) at at the following Web address.
http://imiucca.csi.unimi.it/endomi/micro.html
Thank you Paolo Castano
_____________________________________________________ Prof. Paolo Castano UNIVERSITY OF MILAN INSTITUTE OF HUMAN ANATOMY - CHAIR OF HUMAN ANATOMY FOR PHARMACY Via Mangiagalli, 31 - 20133 Milan (Italy)
Stock 2: 1% Methylene Blue in 1% Borax (Sodium Borate)
Mix stocks 1:1 and store in a syringe with 0.22um filter.
Protocol for 0.5 um EPON or LRGold sections:
1) Transfer sections to drop of ddH2O on a glass slide and warm on a hot plate until water has evaporated.
2) Cover the still-warm sections with stain for 10-30 seconds.
3) Rinse with a stream of ddH2O.
4) Dry sections with a jet of air, then use a KimWhipe to dry the entire slide.
5) Mount in immersion oil.
I've also used this stain on 0.1um cryosections used for orientation prior to cutting cryoultrathins.
} Recently returning to the EM fold after an absence of several years, I } have been unhappy with the staining of LM sections with Toluidine } Blue/Borax or Tolouidine Blue/Basic Fuschin.Buried in a cupboard however } I found a bottle labelled Richardsons Stain which seems to work very } well. However I have no idea whats in it, (its either Toluidine Blue or } Methylene Blue based).I am loathe and embarressed to admit that the } bottle is labelled in my own handwriting, however advancing years and } alzheimers have combined to defeat my memory. Any help would be much } appreciated. } } Peter Smith } AgResearch Wallaceville } Upper Hutt } New Zealand } } smithp-at-agresearch.cri.nz
Happy Staining
Greg Martin Light/Confocal Microscopy and Image Analysis Specialist Biological Imaging Service The Jackson Laboratory TJL Box 43 600 Main Street Bar Harbor. ME 04609
Thanks to Roger Mason for clarifying & verbalizing the distinction between these instruments much better than I did. The "reflecting goniometer" of my experience is (as he describes) a separate, free standing instrument used to measure angles between faces of crystals with well-developed morphology by reflection. So if this is what is wanted, a mineralogy lab (or supply house) is the place to look, as this was a method used in classical mineralogy, probably now largely fallen into disuse. (I was actually taught its use as a preliminary adjunct to space group determination of non-orthogonal crystals.)
Since universal stages, spindle stages, etc are used with polarizing microscopes, it's not surprising that most optical microscopists would think in those terms.
I'm not a mineralogist, but maybe a place to start looking for sources is the Mineralogy Society Website, which has many links http://www.minsocam.org/
Now I'll show my age by saying that in my view, some of these classic methods provided rather elegant and insightful solutions to questions that we now just subdue by brute computational force. If this individual has a neat way to answer a question by measuring crystal morphology, all power to him or her....
Pat Kingman (ex-crystallographer/microscopist now computer jock)
-- ******************************************************************** Pat Kingman Structural Response Team Immediate plans: Code jock....Computer hawk....Cybercadet...
As you say, find out what your source wants. You can improvise something on the cheap (see previous post), or you can rummage, or if already active in x-ray work, you could add reflection to an x-ray goniometer.
FWIW, there exists a fancy spindle stage which will cross over from optical to x-ray work; I know nothing about it but the source, which I found using a search engine on the internet (http:// charles-supper.com/prod01.htm). McCrone's spindle stage is graduated to 10 deg; you would do better with the standard pol stage, even without a vernier. Add a laser pointer if you want. The spindle stage rotates about a horizontal axis on a (usually rotating) stage and is effectively a one-axis goniometer. So is the pol stage.
Leonard Corwin Fort Dodge Animal Health (Analytical Research) Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
A single crystal optical goniometer is available with retired crystallographer for use at the US Geological Survey in Reston, VA. It is under the able guidance of Howard Evans who restored the goniometer to its present excellent condition. In spite of the death of crystallography, mineralogy and petrology at the United States Geological Survey some worthwhile instrumentation survives but only in the hands of Emeritus volunteers.
We invite you to Reston to measure your crystal forms.
Cheers, Gordon
Gordon L. Nord Jr. Scientist Emeritus, Eastern Energy Team 956 National Center Office: 703-648-6745 U. S. Geological Survey FAX: 703-648-6419 Reston, VA 20192 gnord-at-mactem.er.usgs.gov USA
Microscopists, A plant physiologist at our lab would like to stain chilling-injured tissue with a stain specific for oxidative injury. We've tried staining fresh tissue with 10-acetyl-3,7-dihydroxy-2-dodecylphenoxazine and examining it with a confocal microscope without any definitive results. Has anybody been sucessful at this, and if so with what stains/ techniques?
Thanks, Dennis Margosan USDA-ARS 2021 S. Peach Avenue Fresno, CA 93727
} You can boot your Hard drive from a floppy, but unless something is } drastically changed , even using Norton Utilities 3.0 you can not boot up } off a zip. Though Norton Utilites 3.0 lets you reboot a Win 95 system from } a zip AND a floppy. } But you can boot a Mac from a zip.
Bill
} {)))'} {'(((} { Bill Trevarrow, PhD. Zebrafish Facility Director Institute of Neuroscience University of Oregon 1254 Eugene, OR 97403-1254 } {)))'} {'(((} { Off.Tel: (541) 346-4598 Fac. Tel: (541) 346-4512 Fax: (541) 346-4548 e-mail: trevarro-at-uoneuro.uoregon.edu } {)))'} {'(((} {
by mail.unixg.ubc.ca with smtp (Exim 1.71 #1) id 0y724z-0004iv-00; Mon, 23 Feb 1998 09:54:13 -0800
Richardson's stain is 1% Methylene Blue made up in 1% borax, mixed in equal parts with 1% Azure II. You have to filter it to use it. Hope this helps.
Lesley Weston.
On Mon, 23 Feb 1998, Smith, Peter wrote:
} } Recently returning to the EM fold after an absence of several years, I } have been unhappy with the staining of LM sections with Toluidine } Blue/Borax or Tolouidine Blue/Basic Fuschin.Buried in a cupboard however } I found a bottle labelled Richardsons Stain which seems to work very } well. However I have no idea whats in it, (its either Toluidine Blue or } Methylene Blue based).I am loathe and embarressed to admit that the } bottle is labelled in my own handwriting, however advancing years and } alzheimers have combined to defeat my memory. Any help would be much } appreciated. } } Peter Smith } AgResearch Wallaceville } Upper Hutt } New Zealand } } smithp-at-agresearch.cri.nz }
I would like to purchase a copy of N. H. Hartshorne and A. Stuart, Crystals and the Polarising Microscope, Arnold, London, 1969. Please respond directly to me if you have a lead.
Leonard R. Corwin Fort Dodge Animal Health Cyanamid Agricultural Research Center Quaker Bridge & Clarksville Roads PO Box 400 Princeton, NJ 08543-0400 609-716-2278 609-275-5239 fax corwinl-at-pt.cyanamid.com
Hi again, Thanks to those who have replied so far to my posting a couple of days ago, Re: Image slave for S360.
However, a question for those plant TEM'ers out there;
We have a user doing some work on brown macrophyte (seaweed), which is proving very difficult to fix and infiltrate for TEM. It is a very large 'plant' with a VERY thick cell wall and we are interested in structures of Chloroplasts and cell wall junctions. There is also quiet a bit of a polysacharide mucus around the outside of it too. As far as infiltration goes, we have tried Agar 100 (normal epoxy), and Spurrs, with extended infiltration times etc but have had limited success. Literature searhces on this type of thing have been relatively unfruitful to date.
Would love to hear from anyone who deals/has dealt with this stuff before and interested in fixation and infiltration regimes, and any other ideas or relevant references we could look at.
Look forward to hearing from you,
Rich.
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
} A colleague in our Biophysics department makes birefringence } measurements on collagen fibres in paraffin sections of } formaldehyde-fixed specimens. She has come across a 1985 paper } in which related work was done with "Carson's" fixative, } for which no reference or recipe was given. I've not been } able to find this in various texts, ancient & modern. } Thanks in advance for all the replies that will surely } roll in. } John A. Kiernan } Department of Anatomy, } The University of Western Ontario, } LONDON, Canada N6A 5C1 } E-mail: kiernan-at-uwo.ca
A paper was done by FL Carson, comparing paraformaldehyde and formalin in two phosphate buffers to determine an "ideal" fixative for biopsy tissues sent for TEM. the reference for the paper is: American Journal of Clinical Pathology 59:365, 1973
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
The original reference for Modified Millonig's (Carson's fixative) is:
Carson, FL, Martin, JH, Lynn, JA: Formalin fixation for electron microscopy: A re-evaluation. Am J Clin Pathol 59: 365-73, 1973. The pH and milliosmolality more close approximates that of plasma then the usual formulation of neutral buffered formalin. The EM preparations are virtually indistinguishable from those fixed in paraformaldehyde solutions. It has been used as a dual purpose fixative in many labs. There is less lysis with the modified Millonig's than with NBF and the paraffin-embedded tissues are slightly more difficult to section.
Freida Carson
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
With 'El Nino' continuing to make it's presence felt here and water restrictions becoming more stringent we have been reassessing our use of water in the EM Unit. One area of extreme water wastage is our double distilled water setup, perhaps we should be using deionised water instead.
As a result we would be interested in peoples opinions about using distilled water (double distilled) vs deionised water for making up their EM solutions (fixatives, buffers, stains, cytochemical reagents etc).
I guess our attempt is to be a little more 'environmentally correct'!
Would appreciate your thoughts,
Yours,
------------------------------------------------------------ Allan Mitchell Technical Manager South Campus Electron Microscope Unit C/-Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
Fax (03) 479 7254 Phone (03) 479 7301
'The Southernmost EM Unit in the World'
,,, (o o) ------------------oOO-(_)-OOo----------------------------------
I have an early 1980's vintage Zeiss Universal pol. scope with a Diagnostics Instruments adapter and Sony 960 CCD camera. There is an annoying "hotspot" visible in both the live and stored images. This is accentuated in certain instances such as low contrast, slightly crossed polars, etc. The hotspot is almost undetectable in images of samples diffusing a significant amount of light. It is my opinion that the microscope optics are contributing this problem. However, I certainly have an open mind. I have made the following observations: -The microscope is set-up for Kohler illumination. Inserting the diffuser before the lamp housing produces only a slight improvement. -There is virtually no issue when using this camera with the appropriate adapter for our Wild M400 photomacroscope or Olympus Provis Light microscope. -I have tried 2 different Diagnostic Instruments adapters (.6x, .45x) and 3 sets of adapter lenses. In addition, a Dage camera (c-mount) to a "live" monitor shows the same hot spot. -Pol. objectives of that vintage were not flat field. I tried a series of flat field objectives...alas...the field is flat...the hot spot remains. -Local service reps have not been able to implicate the source. -Polaroid and 35mm imaging produces no visible "hot spot". I would be extremely grateful to anyone who could offer suggestions as I have agonized over this for quite some time. Thank you.
Diana Kittleson Pillsbury Technology East 737 Pelham Blvd. St. Paul, MN 55114 dkittleson-at- pillsbury.com
We seek a Technical Coordinator of the Integrated Microscopy Core laboratory in the Department of Cell Biology at Baylor College of Medicine, Houston, Texas. An outstanding career opportunity awaits a skillful applicant with considerable experience in biological light and electron microscopy willing to work with faculty and students. Knowledge of computers and digital microscopy is desired, but not essential. The position entails coordinating a busy core facility, and providing technology and services for a research-intensive faculty. The successful applicant must be experienced in modern specimen preparation, transmission EM and related instrumentation. The laboratory is a well-equipped, state-of-the-art facility (2-TEMs, confocal, deconvolution, two-photon, CCDs, microinjection...).
Salary, working conditions, and cost of living are excellent. Baylor College of Medicine is an Equal Opportunity, Affirmative Action and Equal Access Employer.
Interested applicants should send a CV (email or fax) to:
Michael A. Mancini, Ph.D. B.R. Brinkley, Ph.D. Co-Directors Integrated Microscopy Core Department of Cell Biology Baylor College of Medicine Houston, TX 77030 mancini-at-bcm.tmc.edu 713 798 8952 (voice) 713 790 0545 (fax)
In the two different places where I have used EM, I have had variable experiences. In one situation (East Coast), we took tap water and ran it first through several particle filters (string type, followed by porous ceramic), followed by two large (about the size of tanks of compressed air) deionizer cartridges (one high capacity, one high efficiency) followed by two activated carbon filters and then a millipore filter. This was then autoclaved (for sterility and to eliminate carbon dioxide). This water was normally used for large scale production of interferons in tissue culture systems. The cells grew beautifully and I never had any problems with stain precipitates or fixatives.
Now, (Midwest), the same arrangement gave problems with residual organics that could not be removed by deionization alone. So, we use a much smaller scale system (18 x 4 inch cannisters) consisting of: one string or ceramic type particulate filter, two deionizers (one high capacity followed by a high efficiency) and two activated carbons. These feed into a large still that further purifies down to 1-2 micromhos per cm. This is a 10 fold improvement over deionization/carbon alone. The water is excellent, no precipitates with stains, etc. But distillation is needed in this environment, unfortunately.
My advice, try the deionization/carbon setup first. Do some fixation and staining and if no precipitates occur, you're set. If precipitates, then you'll need to distill. Use some "good" water as a control in this test. We used some water "imported" from Iowa in this test but the source (a person in this case) dried up. Well, the person didn't dry up ........ you know what I mean....
My dry sense of humor .....
} With 'El Nino' continuing to make it's presence felt here and water } restrictions becoming more stringent we have been reassessing our use of } water in the EM Unit. One area of extreme water wastage is our double } distilled water setup, perhaps we should be using deionised water instead. } } As a result we would be interested in peoples opinions about using } distilled water (double distilled) vs deionised water for making up their } EM solutions (fixatives, buffers, stains, cytochemical reagents etc). } } I guess our attempt is to be a little more 'environmentally correct'! } } Would appreciate your thoughts.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I've been asked by a colleague what if anything I knew about the problem of fungi (presumably) that can grow on and etch the front lens of microscope objective lenses, making them useless for any work. This problem may be correlated with tropical or near-tropical environmental conditions.
I couldn't help him at all with any substantive information, but I'll bet this newsgroup can provide answers or leads. Any help would be appreciated. TIA, M. Nesson-- _______________________________________________________________________ Michael Nesson, Ph.D. Department of Biochemistry & Biophysics 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 (541)737-5245 FAX:(541)737-0481 nessonm-at-ucs.orst.edu
We have a Noran Voyager EDXS system attached to a JEOL 2010 TEM. It is one of the older systems and runs under Sunview. We are able to do linescans by setting the endpoints of the line, the elements of interest, image signal and the number of points as well as the dwell time at each point.
One of my colleagues would like to do a continuous linescan between two points. This feature is available in the Link ISIS system under speedmap/linescan but is not available for the Voyager. The beam is repeatedly scanned over the line and data gradually built up until the desired number of scans is carried out.
Does anyone know if it is possible get the Voyager to do this in some way?
Thanks very much in advance.
Colin Veitch
Instrumentation Scientist CSIRO Division of Wool Technology PO Box 21, BELMONT, Vic. 3216. Australia.
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This has been a remarkable thread, full of input from a variety of sources. I think that we have managed to show that there are a variety of possible solutions to the problem, but first, let's consider the options.
If the responses are indicative, and my own experience shows that they are, service from any source is spotty at best. It basically comes down to the service engineer that you will commonly deal with. An SEM is one of the most complex instruments manufactured. It requires a unique individual to not only provide for appropriate service of an SEM but to also provide a customer orientation that can match itself to the expectations of a user. I can confirm that there are those who work for manufacturers who can maintain this balance, as there are those who work for themselves or third party services.
Ken Converse mentioned some of the requirements and costs of maintaining a service engineer, but he was vague in his descriptions. I have seen companies that are very comfortable in justifying spending upwards of $250,000 a year for service engineers, in salary, benefits and travel expenses. Needless to say, the engineer's salary and benefits are the lessor in this equation. At the same time, these same companies will extend additional thousands or tens of thousands of dollars for bringing new hires to the factory for 'factory' training, even if the facility is out of country.
This points to a very inefficient method of training and resource management. At the expenditure levels of some companies, it would make far more sense to provide 4 times the number of service personnel and back those field engineers with a secondary level of expertise of technical specialists and field trainers. I have actually witnessed service organizations whose secondary level of expertise required importing personnel from overseas. A very inefficient use of manpower and a drastic increase in the departmental expenses.
On the other hand, third party services are often taking advantage of the high service costs of the manufacturers. Given the markups, it is very easy to underbid the manufacturers by 10% - 20% and still make a very good living. By undertaking third party service, though, you are opening yourself up to the experience and knowledge of a limited group of field engineers. While the basic engineering of EMs has not changed for years, there can be significant differences in the designs of individual manufacturers. Finding service engineers who can understand and adapt to these differences can be difficult.
The 'HMO' type insurance agencies can, perhaps, offer even greater savings. However, they come with even greater risks. There is no guarantee who will be servicing your instrument. These agencies will pit the manufacturers and the third party sources against each other, and award service to the party that offers the lowest price. There will be no guarantee that the same service personnel, or even the same company, will provide service from call to call.
It basically comes down to how much responsibility you are willing to take for the maintenance of your instrument. There are those who have posted here who do most of the primary service themselves. I basically promote this approach. The more you know about your instrument, the better you can make decisions on the service of it. Being directly tied to the maintenance of your instrument makes you far more informed on the service options available to you.
It is all too common, though, for users to take the easy course of relying on the manufacturer for service. Just as it is all too easy for those management types out there to go for the 'HMO' type service management organizations who offer to cut service costs by attractive amounts. I firmly believe that third party service organizations can offer a reasonable middle ground - consistent service personnel along with flexible contract and billable terms. In most cases, you will be married to your instrument for some time. Try the various alternatives and make up your own mind as to the suitability to your needs. However, think ahead and protect yourself in two ways - demand complete documentation when purchasing - including full schematics, and demand service guarantees when shopping around.
Once again, I am speaking from a biased viewpoint - I am a third party service provider for SEMs and other equipment. However, this being such an esoteric field, I believe that it is especially important that all sources be exploited and supported since the competition can only benefit everyone.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
by haymarket.ed.ac.uk (8.8.7/8.8.7) with ESMTP id IAA27489 for {Microscopy-at-Sparc5.Microscopy.Com} ; Tue, 24 Feb 1998 08:14:43 GMT Received: from SAC005/SpoolDir by ed.sac.ac.uk (Mercury 1.32); 24 Feb 98 08:23:11 GMT Received: from SpoolDir by SAC005 (Mercury 1.32); 24 Feb 98 08:22:58 GMT
Dear all, We are interested in the phosphoglycerate kinase activity in fungi. Does anybody have a staining protocol for the detection of this activity in cells? Armelle Gollotte Biotechnology Department Scottish Agricultural College West Mains Road Edinburgh EH9 3JG Tel: (44) 131 667 1041 Fax: (44) 131 667 2601 a.gollotte-at-ed.sac.ac.uk
} From: Richard Lander {richard.lander-at-stonebow.otago.ac.nz} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Distilled vs Deionised Water? } Date: 24 February 1998 00:11
} With 'El Nino' continuing to make it's presence felt here and water } restrictions becoming more stringent we have been reassessing our use of } water in the EM Unit. One area of extreme water wastage is our double } distilled water setup, perhaps we should be using deionised water instead. } } As a result we would be interested in peoples opinions about using } distilled water (double distilled) vs deionised water for making up their } EM solutions (fixatives, buffers, stains, cytochemical reagents etc). } } I guess our attempt is to be a little more 'environmentally correct'!
I've used deionised and/or reverse osmosis purified water for years. Never had any problems with it. The only time I had to use distilled was many years back when we made our own Gold Colliods. Had to use distilled-deionised water, nothing else worked. Don't know why. At present we use deionised which has been pre-cleaned with a reverse osmosis unit. It comes out of the unit at 18 MgOhm. (At least, that is what the gauge says) But if you need to save water, reverse osmosis runs a lot of raw input water to waste, so it probably isn't any less wasteful than distillation, but running raw water through a deionising resin is expensive on cartridges.
Couldn't you recycle the cooling water to your distillation unit somehow? Through an old EM cooling unit maybe?
I have had a question directed to me regarding water-soluble resins which do not quench fluorescence. Normally the worker cuts cryostat sections but this time the specimens are too small.
The dye used is di-I (whatever that is), it cannot be dehydrated because it is highly soluble in ethanol (and presumably other solvents). All I ever use is epoxies. The desired section thickness is initially around 5-10 microns. Confocal is also being considered. Any tips gratefully accepted!
} As a result we would be interested in peoples opinions about using } distilled water (double distilled) vs deionised water for making up their } EM solutions (fixatives, buffers, stains, cytochemical reagents etc). } ______
My experience has been that from time to time, resin exchange deionized water results in random precipitates on sections. Switching to glass distilled water resolved the problem. I would use ion exchange water with caution.
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
The resins that you mentioned (Spurr, Epon, and Araldite), all need to be removed (etched) prior to staining with either aquous or alcoholic based dyes(there are exceptions). These resins can be etched (removed) with a solution called "Ethoxide". Ethoxide is made as follows:
Make a saturated solution of Sodium Hydroxide in 100% Ethanol and let stand in the dark for at least 48 hours. The solution will turn yellowish when ready.
To make this into a working solution mix equal parts of the above solution and tolulene.
In a fume hood wearing DOUBLE gloves, eye protection, etc. dip the slide gently in the working solution untill you can see that the resin has been removed, dip the slide in 95% ethanol for a minute and then place in a buffer (pH 7.4) for at least 5 minutes prior to staining. You can stain with your normal procedure for the H&E starting with a DI water rinse. I have even done some beautiful Trichromes on sections treated this way.
Some things to be mindful of though........
Do only one slide at a time as it it is very easy to etch out the tissue too.
These sections are much thinner in general than the paraffin sections normally stained with H&E and you may have to increase the staining times a little.
Ethoxide is VERY caustic!!! One drop can do you a lot of damage and pain. I lost a fingernail the second time I used the stuff because of a hole in my glove, it was one of the most painful experiences I've ever had!
There may be a safer and/or easier way to do this but this was what I did before GMA hit the market place back in the early 70's (almost in my 5th decade ;-)). I also have to admit that I haven't done any plastic work in a few years so this could very well be out of date(like me).
I'm going to cross post this to both the Microscopy and the Histology listservs and hope that someone may have a better technique.
-- Begin original message --
} Robert Schoonhoven wrote: } } } } Joan, } } } } Knowing the specific resin would be a help. I have several methods for } H&E staining and they are dependent on the type of plastic the } tissue in embedded in. } } } Robert Schoonhoven } } Dear Dr. Schoonhoven: } } I would be delighted to receive, via e-mail, snail mail or by posting } to the list, your methods for H&E on Spurr, Epon, and Araldite. } Thanks in advance. } } Geoff } -- } *************************************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane Piscataway, NJ 08854 } voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu } *************************************************************** }
-- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
** I'm willing to make the mistakes if someone else is willing to learn from them* *
I would like to improve my double-labelling of vesicles and cytoskeleton in cultured neurons for fluorescence microscopy. For vesicle markers I used monoclonal anti-synaptophysin conjugated with a FITC antibody (whole molecule) and for microfilament markers, I use rhodamine phalloidin.
I have been having problems with the fixation followed by permeabilization schedule. Vesicles do stain and cytoskeleton also does, but vesicles appear to clump as to want to "leave" the cytoskeletal "mesh"-kind of looks like apocrine secretion but these cells are indeed merocrine! We want to see where the vesicles are with respect to exocytosis at different time frames with agonist stimulation.
Does anyone out there have a good protocol for this? I have been using a schedule of a modified Karnovsky's fix for an hour followed by PEG-8000 wash and a 1.25%PEG-8000/0.2%TX-100 in PHEM buffer permeabilization for 10 min. Too long? Thanks in advance for anyones' help.
What do you need to get rid of? Ions, that is salts? Then deionized should be enough.
But! I have rarely found this to be sufficient for microscopy. Organics and non-ionic components are usually more of a problem in water than salts are. I've always have more troubles using deionized water than I have had using distilled. And since distillation may not remove all ions, I've even had to use deionized-distilled.
I don't think water use is the issue here, though. Distillation does consume some water, but it shouldn't be wasting water. It does use more power. Deionization typically uses cartridges, which must be either regenerated (using water) or thrown away. Not counting the manufacture of the resins for the cartridges, or the salts added to your waste water stream. (And deionization consumes water also.)
So I would think distillation is the "better for the environment" solution, as well as being better for your solutions' environment.
Phil
} Hi there again, } } With 'El Nino' continuing to make it's presence felt here and water } restrictions becoming more stringent we have been reassessing our use of } water in the EM Unit. One area of extreme water wastage is our double } distilled water setup, perhaps we should be using deionised water instead. } } As a result we would be interested in peoples opinions about using } distilled water (double distilled) vs deionised water for making up their } EM solutions (fixatives, buffers, stains, cytochemical reagents etc). } } I guess our attempt is to be a little more 'environmentally correct'! } } Would appreciate your thoughts, } } Yours, } } } ------------------------------------------------------------ } Allan Mitchell } Technical Manager } South Campus Electron Microscope Unit } C/-Department of Anatomy and Structural Biology } School of Medical Sciences } P.O. Box 913 } Dunedin } New Zealand } } Fax (03) 479 7254 } Phone (03) 479 7301 } } } 'The Southernmost EM Unit in the World' } } ,,, } (o o) } ------------------oOO-(_)-OOo----------------------------------
I am sure you are right regarding the hot-spot being in the optics of the Zeiss. There are at least 2 possibilities that might cause this. The first is the magnification of the objective. All the lower power Zeiss 1x, 2.5x and even 4x ( and I'm sure most others) had a hot spot in the middle. This can be eliminated to a certain extent, but is due to nature of the beast. Our newer Zeiss axioplan scope had objectives with removable collars on the front to reduce hot spots. Trying polarizing filters (either plane or circular) might help. The second cause is imaging low reflecting surfaces. I am not sure of the exact optical reasons, but it has to do with light reflecting off the specimen and front of the lens. You didn't say whether you had transmitted or reflected light, but each technique has it own problems with low mag hotspots. I suggest that you speak to the Zeiss people in NY. They used to have some really good people to help customers with these type of problems.
Regards, Mike ================================================ Michael L. Boucher Sr. mboucher-at-isd.net WEBPAGE http://www.isd.net/mboucher ================================================
There are stills on the market which far exceed deionized water units in water quality that not only re-use the cooling water in the distillation chamber (thus "preheated") but are also self-cleaning. True there is some "waste" of water (could be saved as tap water to wash dishes I suppose??) I have no financial interest in any companies which produce such stills but have used one made by Weiss Scientific Glass of Beaverton, Oregon which at the time was the only one on the market which recycled the water AND was self-cleaning (water exceeded some of the older "triple-distilled" models in quality). I am unrelated to and have no financial interest in the above mentioned company. Bob Mixon
I was wondering if someone would have some suggestions and also information on where one could acquire Nitrogen standards (except nitrides), in particular some that would be satisfactory to analyze N in silicate material. There is a paper by Ramseyer et al. (1993; J. of Sedimentary Petrology, 63, 1092-1099), that discusses K-NH4-Feldspar in which they are referring to the use of N-doped synthetic cordierite????
Thank you
Cheers
Yves
Yves Thibault Dept. of Earth Sciences B & G Bldg University of Western Ontario London, Ontario, CANADA, N6A 5B7 phone : (519) 661-3186 fax : (519) 661-3198 e-mail : ythibaul-at-julian.uwo.ca
We moved from a lab with deionized water feeding a glass still into a new facility with a reverse osmosis purifier followed by deonization/charcoal tanks, plumbed through the labs with a recirculating system. For EM stains, tissue culture, and molecular biology, we use this water after passing it through a Barnstead NANOpure polishing unit. The RO water is available in all labs and is sufficient for general histology and immunocytochemistry, and EM fixatives. The polishing unit is relatively conveniently located in our tissue culture lab. We have had no problems from the polished water in the 2 years that we have been using it.
Water distillers are no longer allowed to be installed at this campus due to their high energy costs and water inefficiency. My past experiences with non-distilled water systems left me very skeptical of deionization units until we installed our DI/destiller system in 1986. The still was partly to provide backup when the DI system failed. But, the DI system never failed. Our current system has been in use 2 years, so far the only problems have stemmed from lowest bidder installation, not from water quality.
-- Glen MacDonald Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax
} I've been asked by a colleague what if anything I knew about the problem } of fungi (presumably) that can grow on and etch the front lens of } microscope objective lenses, making them useless for any work. } This problem may be correlated with tropical or near-tropical } environmental conditions. } } I couldn't help him at all with any substantive information, but I'll bet } this newsgroup can provide answers or leads.
The Midwest is not the tropics, but we have have two experiences with fungi growing inside of presumably sealed microscope components. In both instances, the fungus grew inside of the prism/beam splitting chamber of a Leica OrthoPlan II. The front-mirrored surfaces were covered with the fungus which then became visible when using the microscope. Besides being annoying, it obscured fine detail. The entire assembly had to be sent back to Germany for cleaning. Leica was most gracious about doing this gratis but now, three years later, the fungus is back. Apparently, it grows on the front surface mirrors which must be replaced.
I have heard that in the tropics, optics (binoculars, or "sealed" optical systems) should be stored in a desiccated environment when not actually being used. Otherwise, fungi get into the "sealed" lens components, grow and etch the glass surfaces or even grow on the lens cement until they completely fill the optics.
My own thinking on this matter would be to periodically "gas" the sensitive components using formaldehyde gas. For example, remove the optics, place in a zip-lock bag with some formalin (38% formaldehyde) and allow gas to permeate the optics. HOWEVER, we need to have some input from microscope manufacturers since the formaldehyde may affect some components inside the lenses. On the other hand, left alone, the lenses will surely be ruined by the fungi.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
There are stills on the market which far exceed deionized water units in water quality that not only re-use the cooling water in the distillation chamber (thus "preheated") but are also self-cleaning. True there is some "waste" of water (could be saved as tap water to wash dishes I suppose??) I have no financial interest in any companies which produce such stills but have used one made by Weiss Scientific Glass of Beaverton, Oregon which at the time was the only one on the market which recycled the water AND was self-cleaning (water exceeded some of the older "triple-distilled" models in quality). I am unrelated to and have no financial interest in the above mentioned company. Bob Mixon
} I have heard that in the tropics, optics (binoculars, or "sealed" optical } systems) should be stored in a desiccated environment when not actually } being used. Otherwise, fungi get into the "sealed" lens components, grow } and etch the glass surfaces or even grow on the lens cement until they } completely fill the optics.
Well, I am in the tropics, but we are currently having a drought because all you U.S. coastline people have stolen our water!
} My own thinking on this matter would be to periodically "gas" the sensitive } components using formaldehyde gas. For example, remove the optics, place in } a zip-lock bag with some formalin (38% formaldehyde) and allow gas to } permeate the optics. HOWEVER, we need to have some input from microscope } manufacturers since the formaldehyde may affect some components inside the } lenses. On the other hand, left alone, the lenses will surely be ruined by } the fungi.
I would love to find out if this would work! Once discovered, keeping the affected glass optics dry prevents spread of the fungus, but it gets going again if rehydrated, I think.
My experiences with containers and humidity has led me to a number of conclusions. First, zipper-type plastic bags don't really keep out moisture all that well for very long. Don't rely on them. And I'd bet the formaldehyde vapors would permeate OUT of the bag, as well, so don't do this at home. I am ready to stand corrected, of course.
Secondly, Parafilm does an amazing job of sealing things up. It took me a long time to figure out that Parafilm wrapped around the lid of any old jar will keep indicator dessicant blue for years. I trust it, now. My camera equipment is in a peanut butter jar with dessicant and Parafilmed. (Anyone remember my rant about peanut butter jars for storage a year or so ago?)
Next, of all the commercial sealable food storage containers out there, I have found Tupperware brand to be the best at sealing out humidity. I have had a couple of hilarious Tupperware parties for scientists where the poor salesperson was bewildered by our refusal to play the games, and our discussion of pathology and oceanography. It's easier to go to the local supermarket for Rubbermaid or equal brand, but they just don't do the job.
And something I found out the *hard* way - Drierite (CaSO4) can be corrosive, so we have switched to silica gel.
We sometimes put a "head" of N2 gas before sealing things.
My $.04 (twice as long as it should be).
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
At 06:20 PM 2/23/98 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I eventually had to replace lens elements in a high-priced macro lens. Other than trying to keep the lenses in a low-humidity environment (i.e., storing in containers with silica gels), I'm not personally aware of any preventative for fungus growing on lens surfaces or lens coatings. Nor, unfortunately, am I aware of any cheap fixes.
Maybe some other people have better solutions.
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
We have had a couple of experiences with distilled vs deionised water issues. First when using glass distilled water our isolations of sperm cells from the pollen of Plumbago worked just fine. The university went to a campus wide Reverse Osmosis (RO) system and our isolations began to fail. Now we use RO water that has been deionised the isolations again work just fine. We are using a single high-efficiency DI filter cartridge which delivers water in the 18 MgOhm range according to the meter on the unit. Another related issue concerns the plumbing for the distilled/RO water supplied to the building. The taps that were installed were made of PVC which in itself is fine, but the anti-siphon valves that were installed were metal and incorrect for a distilled/RO water tap. We were informed that the lines should have had anti-siphon valves for DI/RO water installed and that the anti-siphon valves we had on there were probably leaching elements into the DI/RO water. The replacement taps (several of ours were broken) were of the metal variety and I was informed that they were tin lined and very expensive (thankfully replaced under repair to building!). So to make a long story short, we have had no troubles since going to DI water although we are deionizing water that has already been through reverse osmosis. For some reason the reverse osmosis water alone is not good enough for our purposes. As a secondary concern the way that the water is delivered may be an issue. I do not know if the anti-siphon valves were a problem or not as our isolations worked fine even though we had anti-siphon valves in place that were not "proper" for DI/RO water taps. As far as EM solutions - we use deionized water and have seen no problems. Greg -- ======================================================== Greg Strout Electron Microscopist, University of Oklahoma e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not neccessarily those of the University of Oklahoma ========================================================
I am intersted in finding out how many of the clinical EM labs there are out there that are currently using digital imaging to process their EM micrographs. If digital imaging is being used, how is it being used? (ie, scanning negatives, or using a digital camera attached to the TEM, or any other adaptations.) I would also be interested in knowing what types of equipment are being used, ie scanners, cameras, sofware, storage, etc.
Any information would be appreciated! I can be contacted at: hcmcemlab-at-sprintmail.com
Thanks for your help! Kerstin Halverson, M.S., BEMT (MSA) Hennepin County Medical Center EM Lab
Hi ! Is anyone aware of a good manual detailing microscopy techniques using membrane filters ? I work for a filter company and I am constantly being asked for recommendations/protocols for visualizing cells etc captured on microporous membranes. Have you ever run across such a book ? Your assistance is GREATLY appreciated ! Most of my clientrs are interested in light microscopy/general staining and occasionally SEM and TEM techniques.
Lisa Vickers Millipore Corporation 80 Ashby Rd Bedford, Ma. 02173
Be warned that this problem not only occurs in tropical, humid climates, Wellington , New Zealand can hardly be called tropical. We experienced this problem on a surgical microscope .The problem appeared over winter/early spring when the weather was damp but hardly tropical. Luckily we caught the problem before any permenant damage was done. The microscope which is not used to often is now stored in a dry atmosphere and inspected regularly as once the problem has occurred it is far more likely to reoccur (residual spores etc)
Could anyone please point me in the right direction with regard to the preparation of an Ethylene Vinyl Acetate film for ultrathin sectioning and TEM investigation.
We are a biological lab used to working with muscles, livers, brains and the like; this material is a new challenge for us.
The questions I would like answered include;
- does it need fixing before processing, if so fix in what (Glut, OsO4, buffer??)
- do you dehydrate in solvent ?
- what resin should I use ?
A protocol or references would be greatly appreciated.
Many thanks
Allan Mitchell } } ------------------------------------------------------------ } Allan Mitchell } Technical Manager } South Campus Electron Microscope Unit } C/-Department of Anatomy and Structural Biology } School of Medical Sciences } P.O. Box 913 } Dunedin } New Zealand } } Fax (03) 479 7254 } Phone (03) 479 7301 } } } 'The Southernmost EM Unit in the World' } } ,,, } (o o) } ------------------oOO-(_)-OOo---------------------------------- } } ----------------------------------------------------------------------- } Richard Lander } Electron Microscope Technician } South Campus Electron Microscope Unit } Otago School of Medical Sciences } P.O. Box 913 } Dunedin } New Zealand. } Tel. National 03 479 7301 Fax. National 03 479 7254 } } "Southernmost EM Unit in the World!" } ------------------------------------------------------------------------ }
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
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All-
We are currently seeking a source for single-crystal YAG scintillators to use in a high-voltage TEM camera. Our current YAG is a 25mm disk that appears to be radiation damaging after several years use at 1.5MeV. Ideally, we need a larger YAG disk, up to 50mm diameter, with optically-flat surfaces and 0.1mm thickness. If necessary, we should be able to have a thicker disk, say 0.5mm, ground locally to the required 100 micron. Any leads to suppliers or tips on experiences of YAGs vs phosphors would be welcome.
by yowie.cc.uq.edu.au (8.8.8/8.8.8) with SMTP id MAA00057; Wed, 25 Feb 1998 12:45:10 +1000 (GMT+1000)
} -Randy it is a very common problem for fungi to get into the layers that make up a lens. In my experience it is incurable,(Anyone please correcr me if there is now something that I don't know about). In my 25+ years in the humid tropics I never had the fungi problem while other people around me did. I have always attributed this to the fact that I never covered my microscope, prefering to let the air circulate around it. I know that it is considered sacrilage not to cover your microscope but my microscopes have certainly outlasted numerous others and never had the dreaded fungi.
Christine Lee, Senior Scientific Officer, Veterinary Pathology, University of Queensland.
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Interesting supposition. However, given that I have been a Service Engineer, Technical Specialist and Service Manager for some 25 years, I'm not sure that you are quite on mark.
I also don't think that most here would accept this as a reasonable response in what has otherwise been an interesting thread.
I do find it interesting that you have not taken any other part in this discussion. I do remember your fellow Philips employee Clay Jordan's remarks though. I think you should put him back on the line, at least he gave an appearance of some neural activity.
My apologies to all.
} Once again Allen has shown his negative bias against SEs (Service } Engineers). With this type of bias all you get is BS. } } John Beardslee } FEI / Philips
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Mike O'Keefe wrote: ================================================== We are currently seeking a source for single-crystal YAG scintillators to use in a high-voltage TEM camera. Our current YAG is a 25mm disk that appears to be radiation damaging after several years use at 1.5MeV. Ideally, we need a larger YAG disk, up to 50mm diameter, with optically- flat surfaces and 0.1mm thickness. If necessary, we should be able to have a thicker disk, say 0.5mm, ground locally to the required 100 micron. Any leads to suppliers or tips on experiences of YAGs vs phosphors would be welcome. ===================================================== The problem is less a matter of making it that thin than it is in the transport of getting it into the hands of the customer without it breaking. We could in theory supply such a disc however what we would not be able to do is to guarantee that it would not arrive broken and also, guarantee that you could get it installed in your system without its breaking once in your possession. If we were talking about something of nominal value, it would be different but these things are not cheap either.
Supplying you with the disc that is on the order of 0.5 mm is no problem, contact me off line and we can talk about it. However, I would urge caution because at 100 um, the disc will be extremely fragile and only those with the most magic of fingers I am told can do it. But even then we have been told about the polished disc shattering when installation was attempted.
I am not sure there are any simple answers here but the problem is one that has our attention.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I expect you will be inundated with responses from US suppliers but over the years we have obtained a range of YAG crystals from Agar Scientific in the UK. Phone (44) 1279 813591 or fax (44) 1279 815106. They are obviously the middle men but seem to know where to go for most things. Usual disclaimers - I'm a happy customer.
Ron ========================================================================== = Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ========================================================================== ==
The LM maintenance section at our local (latitude 19) University years ago researched that topic. Most of the microscopes are kept in airconditioning and then no precautions are required but for the Coral Sea island research station they enclose a teaspoon of paraformaldehyde powder in a small paper bag within the microscope case. This protects the microscope for a year or two and apparently has no ill effect on lenses and the mechanical parts. Another alternative are the "plug-in and forget" new desiccating cabinets. They keep relative humidity below 20%; a true innovation. I must declare an interest: ProSciTech sells these. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} Microscopists: } } I've been asked by a colleague what if anything I knew about the problem } of fungi (presumably) that can grow on and etch the front lens of } microscope objective lenses, making them useless for any work. } This problem may be correlated with tropical or near-tropical } environmental conditions. } } I couldn't help him at all with any substantive information, but I'll bet } this newsgroup can provide answers or leads. } Any help would be appreciated. } TIA, } M. Nesson-- } _______________________________________________________________________ } Michael Nesson, Ph.D. Department of Biochemistry & Biophysics } 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 } (541)737-5245 FAX:(541)737-0481 nessonm-at-ucs.orst.edu } } }
We have encountered the same type of fungus here in New York City, growing on the surface of a diamond knife. As the fungus approached the sharp edge of the knife, that portion was ruined. The precipitating cause seemed to be a knife storage box which was air tight, and our own carelessness. If the knife was placed into the box wet, and the box shut before the knife had dried, the fungus grew. Over time, the fungus eventually attacked the entire knife surface, despite efforts to dry the knife after each use. David Hall Department of Neuroscience 1410 Pelham Parkway Albert Einstein College of Medicine Bronx, NY 10461
Credits to all that have contributed to this thread;
But does anyone know the name of this type of fungus? What type of glass does it grow on and what component of the glass is so attractive to it?
Before my business became overwhelmingly active I used to restore old optical systems for historical exhibition. I no longer have time for that now but after years of seeing great optics ruined by this fungus I am glad others have raised this topic. Hopefully I'll learn more about it.
Hi Could you explain me why you don`t see those hot spots on photomicrography? Even if the field observed on them are smaller, those spots should also be there? I still think they are related to C-Mount used and internal reflection of them. also the front glass covering the CMOS has something to do with it. Norm At 10:11 24/02/98 +0000, Mike Boucher wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I needed some primary cells for lab in a hurry, so I trypsinized= my unsuspecting technician...seriously, the technician in our EM= Facility has moved to California so there is an opening which must= be filled. The official description follows:
EM TECHNICIAN- The Department of Biology at Western Kentucky University= is accepting applications for a full-time, permanent Electron Microscopy= Technician position available July 1, 1998. The major responsibility= of this position is to oversee activities associated with the= multi-disciplinary user EM Facility. The Facility houses two JEOL= 100B TEMs, one JEOL 5400LV SEM with EDS, a darkroom, and biological= sample preparation equipment. The technician will be responsible= for the day-to-day operations of the Facility including maintenance= of the instruments and trouble-shooting, sample preparations, inventory= of equipment and supplies, training of users, and providing research= support for faculty, students, and outside users of the Facility.= The person hired will be encouraged to assist in obtaining outside= support for the Facility through contracts. The technician will= report directly to the Facility Director. The qualified candidate= will have a B.S. degree (M. S. preferred) and at least two years= experience in aspects of electron microscopy. Additional duties= will be assigned and may include supervising laboratory preparations= for cell and molecular biology courses and full support for the= darkroom. Send letter of application, curriculum vitae and three= letters of reference to: Dr. Laura S. Rhoads, Department of Biology,= Western Kentucky University, 1 Big Red Way, Bowling Green, KY 42101-3576.= Screening of applications will begin April 10, 1998. Questions may= be addressed to laura.rhoads-at-wku.edu. Women and minorities are especially= encouraged to apply. WKU is an affirmative action/equal opportunity= employer.
Please post this message for your colleagues. Thank you.
************************************************************ It's true- the inmates ARE running the asylum... ************************************************************ Laura Rhoads Electron Microscopy Facility Director Department of Biology Western Kentucky University 1 Big Red Way Bowling Green, KY 42101-3576
Hi! Anybody know anything about Sputter Coaters? Having trouble with EMSCOPE model SC500A, Auto sequencing not operating correctly. Any ideas?? Peter Toronto
This message is in MIME format. The first part should be readable text, while the remaining parts are likely unreadable without MIME-aware tools. Send mail to mime-at-docserver.cac.washington.edu for more info.
Are you people *sure* this problem is biological, caused by a fungus? Might it not be a devitrification of the glass? I worked several years ago with a student in Art Conservation from the nearby Winterthur museum. Her specialty was lockets which contained pictures behind glass. The glass in these often had the exact problem you describe. Although the stuff on the surface of the glass looked like a growth of some kind, it was actually a gel of glass material in water. She told me this is common knowledge in her field, and is in all the textbooks. One giveaway that this is something done by the glass is that it only happens to flint glass, not to crown. You'd think a fungus would do it the other way around; what would preferentially grow on a material that is 10 or more percent lead? Is there a newsgroup or mailing list for optical repair to which we could submit this question? If there is no fungus, then fumigation won't help, and dessication is the only preventative.
Robert Wieland wieland-at-me.udel.edu Electron Microscope Specialist University of Delaware Neither Yankee nor Dixie, east of the Mason-Dixon line (look it up). You can't go faster than light, you can't get colder than absolute zero, and you can't help somebody by not telling them the truth.
First Announcement (SHORT VERSION, only ASCII-Text) NOTE:=20 a SECOND (extended) VERSION (extended ASCII-Text only) will be posted = soon to this listserver. a THIRD VERSION, containing only a WORD-attachment will be forwarded = later.=20 Apologize for that procedure but we would like to have posted this = information to ALL members of this List-Server community (also those members, = accepting only e-mails by ASCII-text without attachments and without = Internet-connections).
=20 Dear colleague(s), as a member of the Scientific Committee I would like to announce=20
SIXTH INTERNATIONAL CONFERENCE AND WORKSHOP ON
MOLECULAR MORPHOLOGY
to be held at: SALZBURG, Austria (Europe), OCTOBER 5-9, 1998
ORGANIZING FACULTIES: The University of Salzburg, Medical Research Coordination Center Salzburg General Hospital (St. Johanns-Spital LKA),=20 Institute of Pathological Anatomy The International Society of Molecular Morphology, Oklahoma City, OK, = USA The Cleveland Clinic Foundation, Cleveland, OH, USA
TOPICS will include: } in-situ { PCR; super-sensitive } in-situ { hybridization and catalyzed = reporter deposition (CARD) technology; automated single gene copy = staining; immunohistopathology and tumor markers; telepathology; = } in-situ {-techniques in pathology; peptide technologies; multicolor = fluorescence; flow cytometry; autometallography; immunogold-silver = staining and Nanogold; EM immuno- and } in-situ { labeling; permanent = multiple immunostaining; organotypic brain slices; electrophysiology; = tumor viruses; cytoskeleton; corrosion casts; computer image analysis; = DNA ploidy; luminescence analysis; argyrophilia and = argentaffinity.......
PLENARY LECTURERS: Julia M. POLAK (London, U.K.): Nitric Oxide in Health and Disease Virginia M. ANDERSON (Brooklyn, N.Y.): Impact of Molecular Morphology on = the New Millenium Jules ELIAS (New York, N.Y.): Molecular } in-situ { techniques in = Pathology and Biology ............ INVITED WORLD-CLASS LECTURERS will include:
Omar BAGASRA (Philadelphia, P.A., USA) Gorm DANSCHER (Aarhus, Denmark) Lawrence DeBAULT (Oklahoma City, OK, USA) Ursula FALKMER (Trondheim, Norway) Gian-Luca FERRI (Cagliari, Italy) Lars GRIMELIUS (Uppsala, Sweden) Jiang GU (Laurel, N.J., USA) James HAINFELD (Upton, N.Y., USA) Victor SMALL (Salzburg, Austria) S. K. TANG (Kew, Australia) Raymond TUBBS (Cleveland, OH, USA)=20
WET-WORKSHOPS and DEMONSTRATIONS are planned: see forthcoming Announcement II (long version), this List-server
(a limited number of seats in the wet-workshops is available on a "first = come-first serve"-basis)=20
Congress fee will be Austrian Schilling (ATS) 5,000.- (approx. US$ = 385.-) including all lectures, coffee, tea and cakes.
For FURTHER INFORMATIONS and for your convenience an
INTERNET-WEBPAGE is available (completed by March, 1st, 1998) at:
} } } } http://www.kongress.at/IMMC { { { {
For further informations, pre-registration and seat-reservation in = special workshops, please contact:
Prof. Dr. Gerhard W. HACKER Medical Research Coordination Center LKA SALZBURG c/o Dept. Pathol.-Anatomy Muellner Hauptstrasse 48 A-5020 SALZBURG, AUSTRIA Phone: ++43++662-4482-4730 ext., Fax: ++43++662-4482-882 ext., or:
e-mail: g.hacker-at-lkasbg.gv.at
Thank you very much for your attention, best regards to all the List-members
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
First Announcement (SECOND -extended- VERSION ( ASCII-Text only)=20 NOTE:=20 a THIRD VERSION, containing only a WORD-attachment will be forwarded = later.=20 Apologize for that procedure but we would like to have posted this = information to ALL members of this List-Server community (also those members, = accepting only e-mails by ASCII-text without attachments and without = Internet-connections).
=20 Dear colleague(s), as a member of the Scientific Committee I would like to announce=20
SIXTH INTERNATIONAL CONFERENCE AND WORKSHOP ON
MOLECULAR MORPHOLOGY
to be held at: SALZBURG, Austria (Europe), OCTOBER 5-9, 1998
ORGANIZING FACULTIES: The University of Salzburg, Medical Research Coordination Center Salzburg General Hospital (St. Johanns-Spital LKA),=20 Institute of Pathological Anatomy The International Society of Molecular Morphology, Oklahoma City, OK, = USA The Cleveland Clinic Foundation, Cleveland, OH, USA
For your convenience an INTERNET-WEBPAGE is available=20 (completed by March 1st, 1998) at: } } } http://www.kongress.at/IMMC { { {
SCOPE: State-of-the-art conference of today's latest morphological } in = situ { molecular techniques for LM and EM, each represented by = world-class leaders. Aimed for beginners as well as for advanced = scientists who will find first-hand information, the possibility of = fruitful discussion, as well as wet-workshop training
TOPICS will include: } in situ { PCR; super-sensitive } in situ { hybridization and catalyzed = reporter deposition (CARD) technology; automated single gene copy = staining; immunohistopathology & tumor markers; telepathology; } in situ { = techniques in pathology; peptide technologies; multicolor fluorescence; = flow cytometry; autometallography; immunogold-silver staining and = Nanogold; EM-Immuno- and } in situ { labeling; permanent multiple = immunostaining; organotypic brain slices; electrophysiology; tumor = viruses; cytoskeleton; corrosion casts; computer image analysis; = DNA-ploidy; luminescence analysis; argyrophilia and argentaffinity.
PLENARY LECTURES: Julia M. POLAK (London, UK): Nitric Oxide in Health and Disease Virginia M. ANDERSON (Brooklyn, N.Y.): Impact of Molecular Pathology on the New Millenium Jules ELIAS (New York, N.Y.): Molecular } in-situ { Techniques in = Pathology and Biology Jules ELIAS will also give a series of sunrise-lectures introducing = molecular biology to medical professionals
CONGRESS FEE: Austrian Schilling (ATS) 5,000.- (approx. US-$ 385.-) INCLUDING: all lectures, coffee, tea and cakes
MAIN LECTURES: at now, 27 "invited" main lectures given by renowned = scientists in their field are scheduled: INVITED WORLD-CLASS LECTURERS will include:
Omar BAGASRA (Philadelphia, P.A., USA): } in-situ { PCR Gorm DANSCHER (Aarhus, Denmark): Autometallography=20 Lawrence DeBAULT (Oklahoma City, OK, USA): } in-situ { transcription Ursula FALKMER (Trondheim, Norway): DNA ploidy and IHC Gian-Luca FERRI (Cagliari, Italy): Multicolor = fluorescence Lars GRIMELIUS (Uppsala, Sweden): Endocrine Pathology and = Silver Jiang GU (Laurel, N.J., USA): = Telepathology James HAINFELD (Upton, N.Y., USA): Nanogold and undecagold Victor SMALL (Salzburg, Austria): Cytoskeleton S. K. TANG (Kew, Australia): = "Thin-Prep"-Smears Raymond TUBBS (Cleveland, OH, USA): Automated CARD-ISH and Flow Cytometry of lymphoproliferative disorders
WORK-SHOPS and DEMONSTRATIONS (additional, small fees): besides a series of parallel wet-workshops most important to the = interested community: } in-situ { PCR; Manual ultrasensitive } in situ { hybridization; = Nanogold-silver-staining for immunohistochemistry and DNA/RNA detection; = Immunogold-silver staining for Electron Microscopy; CARD-} in situ { = hybridization for LM & Electron Microscopy; Automated single gene copy = DNA and RNA detection; multicolor fluorescence; Interactive DNA ploidy analysis; Telepathology; = luminescence analysis; Calcium analysis using confocal laser microscopy; = .........
For further information, pre-registration, seat-reservation in special = workshops,=20 visit the INTERNET-WEBPAGEs (completed by March 1st, 1998) at:
} } } http://www.kongress.at/IMMC { { { =20 or please contact: Prof. Dr. Gerhard W. HACKER,=20 Medical Research Coordination Center, LKA Salzburg Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA Phone: ++43++ 662-4482-4730 Fax: ++43++ 662-4482-882 or, preferentially by e-mail: g.hacker-at-lkasbg.gv.at
Yours respectfully,
Dr. Wolfgang MUSS Department of Pathology, LKA, EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
Stephan Helfer Royal Botanic Garden Inverleith Row Edinburgh EH3 5LR Scotland UK
} Date: Wed, 25 Feb 1998 07:34:09 -0500 } To: microscopy-at-sparc5.microscopy.com } From: "Dr. David Hall" {hall-at-aecom.yu.edu} } Subject: lens-eating fungi
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We have encountered the same type of fungus here in New York City, growing } on the surface of a diamond knife. As the fungus approached the sharp edge } of the knife, that portion was ruined. The precipitating cause seemed to } be a knife storage box which was air tight, and our own carelessness. If } the knife was placed into the box wet, and the box shut before the knife } had dried, the fungus grew. Over time, the fungus eventually attacked the } entire knife surface, despite efforts to dry the knife after each use. } David Hall } Department of Neuroscience } 1410 Pelham Parkway } Albert Einstein College of Medicine } Bronx, NY 10461 } } phone (718) 430-2195 FAX (718) 430-8821 }
As chemists, we use multiple-cartridge deionizers to prepare "pure" water for analytical purposes. The resistivity does not tell you if the water will grow cells or do other things. Apart from functionality, you may wish to find out how noisy the units are in operation if you will be nearby, especially if they periodically turn on a recycling pump. In my experience, Barnstead units are much quieter than Millipore ones.
Leonard Corwin Fort Dodge Animal Health (Analytical Research) Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
Hello from northeast Ohio, An ethylene vinyl acetate copolymer can be of varying hardness depending on the percentage of each component - i.e., 20-30% vinyl acetate produces a rubbery material while a concentration above 75% produces a very rigid material. Most of the polymers are rubbery. In any case, the ethylene component has a low Tg (glass transition temperature) and the polymer should be cryo-ultramicrotomed at temperatures below -70oC. You should be able to stain the sections afterwards using a saponification/OsO4 technique. I am assuming that you are trying to see the different phases. Alternatively, very small pieces of the plastic could be immersed in fresh RuO4, embedded in plastic and ultramicrotomed. Good luck, this polymer is not that easy to stain. Vicky (Bauer) Bryg Microscopist Ferro Corp. (216) 641-8585 x6613 vbauer-at-ferro.geis.com (ignore the long address above)
We're looking desperately for anyone who can help with software for an EDAX 9100--the version we need is version 2.2 or 2.3 with the FT0 and FT1 designation on single-sided 8 inch floppies. Any help or leads would be greatly appreciated. Feel free to contact me off-list at vrieck-at-spectra-inc.com. TIA,
Vern Rieck Principal Engineer Spectra, Inc. Hanover, NH 603-643-4390
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Job Posting:
Sr. R&D Scientist, Department of Microscopy and Image Analysis, Helene Curtis-Unilever Home and Personal Care USA. Rolling Meadows, Illinois
Experienced microscopist with B.S, M.S. or PhD in biology or chemistry, with 5 to 10 years microscopy work experience. Individual must be able to work independently, problem solve, and have the capability and initiative to run the microscopy facility, perform new technique development and supervise microscopy personnel. Qualified individual should have proficiency in TEM, SEM, EDS, light microscopy, microtomy, immunohistochemistry, and cryomicroscopy. Salary is competitive.
Please respond to this posting to : Joanne M. Crudele, Manager, Microscopy and Image Analysis, Helene Curtis, 3100 Golf Rd., Rolling Meadows, Illinois 60008. (847) 734-3712 or fax (847) 734-3686.
with smtp id {m0y7meC-00035RC-at-www.qtmsys.com} (Debian /\oo/\ Smail3.1.29.1 #29.33); Wed, 25 Feb 98 14:37 EST Message-Id: {3.0.32.19980225143807.00688fc0-at-blackhole.qtmsys.com} X-Sender: dbailey-at-blackhole.qtmsys.com X-Mailer: Windows Eudora Pro Version 3.0 (32)
I don't know much about that, but Fungi grow also in Arsen-solutions and many HIGHLY toxic materials - probably an interesting field for research. I've also seen pictures in a german textbook, where a fungus was shown growing on "ordinary" glass, leaving behind it etching traces where the hyphae were grown. And since people wrote that fumigation helps, it should also be a living thing within the microscopes?
it was written in the list: Are you people *sure* this problem is biological, caused by a fungus? Might it not be a devitrification of the glass? ..... what would preferentially grow on a material that is 10 or more percent lead? Is there a newsgroup or mailing list for optical repair to which we could submit this question? If there is no fungus, then fumigation won't help, and dessication is the only preventative.
Dr. Arthur Schuessler University of Heidelberg Zellenlehre Im Neuenheimer Feld 230 D-69120 Heidelberg Germany
One of the fungi that does this is Aureobasidium pullulans, it is also seen often growing on tile grout in bathrooms. My best guess it is using carbon in the coatings or cements, or thin oil films that form from the air.
Russ
} Subject: Re: Lens-eating Fungi?? } Date sent: Wed, 25 Feb 98 09:47:11 -0400 } From: Dan {dan-at-bioptechs.com} } To: {Microscopy-at-Sparc5.Microscopy.Com}
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Credits to all that have contributed to this thread; } } But does anyone know the name of this type of fungus? } What type of glass does it grow on and what component of the glass is so } attractive to it? } } Before my business became overwhelmingly active I used to restore old } optical systems for historical exhibition. } I no longer have time for that now but after years of seeing great optics } ruined by this fungus I am glad others have raised this topic. } Hopefully I'll learn more about it. } } Dan } } } } } Russell N. Spear Sr. Research Specialist Dept. of Plant Pathology Univ. of Wisconsin-Madison
Apparently, the last salary survey for microscopists, actually the em variety, was published in 1984 by EMSA. I have had several direct ( to me) and a few responses over this listserver stating they were interested in my findings. However, I came up with zip (although more people seem interested in zip drives then this topic). It would seem the time is ripe for another survey, maybe by same organization or one of the other publications that cover microscopy. With most members of MSA setup with email and thus could reply to a questionnaire electronically, it would seem to be less work then the last time. Just a suggestion and my two centavos worth. Hank Adams Electron Microscopy Lab New Mexico State University Las Cruces,NM 88003 phone: 505-6463600 fax: 505-6465665
Please, unsubscribe Thanks, Lic. Veronica Campanucci -------------------------------------------------------------------------------- Lic. Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
Looking for recommendations and/or references for reader's favorite methods for rotary replication of purified proteins. Any and all details of the method are greatly appreciated. I will be using a Balzers 400T to generate the replicas.
TIA, Steve Samuelsson
Steve Samuelsson, Ph.D. Procter & Gamble Pharmaceuticals, Inc. PO Box 8006 8700 Mason-Montgomery Road Mason, OH. 45040-8006 (513) 622-1753 office (513) 622-1752 lab (513) 622-1196 fax samuelsson.sj-at-pg.com
I would like to thank all of you for your responses about photo-quality printers. I will post a summary of responses in a day or two.
We have heard a rumor that the Codonics dye-sub printer is powered by the Kodak printer engine. Can anyone comment on this?
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Group - With Hank Adam's suggestion on this subject, I will take on the effort to complete a proper salary survey for microscopy with our publication "Microscopy Today." In that we have some 8,000 microscopy readers in the U.S., the publication (plus this vehicle) would seem ideal for the effort. What I first need is any help YOU each can allow to the most effective format. Categories, education, experience, etc. The results, of course, will be available on this medium (and our publication) at no charge. If you are interested in this subject, kindly do assist me in the creation of a proper and effective format. It is only with a good format will the survey results truly be of value. Regards to all, Don Grimes, Microscopy Today
Here is a summary of peoples ideas re: distilled water vs de-ionised. Thanks to all who responded. **************************** John Bozzola wrote: In the two different places where I have used EM, I have had variable experiences. In one situation (East Coast), we took tap water and ran it first through several particle filters (string type, followed by porous ceramic), followed by two large (about the size of tanks of compressed air) deionizer cartridges (one high capacity, one high efficiency) followed by two activated carbon filters and then a millipore filter. This was then autoclaved (for sterility and to eliminate carbon dioxide). This water was normally used for large scale production of interferons in tissue culture systems. The cells grew beautifully and I never had any problems with stain precipitates or fixatives. Now, (Midwest), the same arrangement gave problems with residual organics that could not be removed by deionization alone. So, we use a much smaller scale system (18 x 4 inch cannisters) consisting of: one string or ceramic type particulate filter, two deionizers (one high capacity followed by a high efficiency) and two activated carbons. These feed into a large still that further purifies down to 1-2 micromhos per cm. This is a 10 fold improvement over deionization/carbon alone. The water is excellent, no precipitates with stains, etc. But distillation is needed in this environment, unfortunately. My advice, try the deionization/carbon setup first. Do some fixation and staining and if no precipitates occur, you're set. If precipitates, then you'll need to distill. Use some "good" water as a control in this test. We used some water "imported" from Iowa in this test but the source (a person in this case) dried up. Well, the person didn't dry up ........ you know what I mean....
******************************* Julian Smith wrote:
I'm not sure it results in a net savings, but I use the following system and like it very much: 1. Spun-fiber 9=B5m filter 2. RO cartridge (80-liter/day system, often sold to private homes at the beaches here, to remove sulfide and iron) 3. Mixed-bed DI 4. Storage jug with recirculating pump for mixed-bed DI 5. Milli-Q system
I use the water in the storage jug for final rinse on dishes and for all non-critical solution mixing (I make my EM fixatives up in it, for instance). It's about the same as our house distilled, about 8=B5S. I use the Milli-Q water for all critical stuff, ranging from Locke&Krishnan silver staining to PCR and protein electrophoresis. No problems so far.
The RO cartridge has a rejection ratio of about 10:1 though, so for every 10 liters of water in the jug, I'm putting about 90 down the drain.
HTH Julian
************************** =46rom: "Brian G. Demczyk" {demczyk-at-erxindy.rl.plh.af.mil}
There is no problem with using deionized water as long as a bit of "tapwater" is added, as well. Typically a cup or two in a recirculation unit filled with deion water is sufficient.
*************************** =46rom: Donald Lovett {lovett-at-TCNJ.EDU}
My experience has been that from time to time, resin exchange deionized water results in random precipitates on sections. Switching to glass distilled water resolved the problem. I would use ion exchange water with caution.
******************************* =46rom: Stephen Griffiths {s.griffiths-at-ucl.ac.uk} I've used deionised and/or reverse osmosis purified water for years. Never had any problems with it. The only time I had to use distilled was many years back when we made our own Gold Colliods. Had to use distilled-deionised water, nothing else worked. Don't know why. At present we use deionised which has been pre-cleaned with a reverse osmosis unit. It comes out of the unit at 18 MgOhm. (At least, that is what the gauge says) But if you need to save water, reverse osmosis runs a lot of raw input water to waste, so it probably isn't any less wasteful than distillation, but running raw water through a deionising resin is expensive on cartridges.
Couldn't you recycle the cooling water to your distillation unit somehow? Through an old EM cooling unit maybe?
What do you need to get rid of? Ions, that is salts? Then deionized should be enough.
But! I have rarely found this to be sufficient for microscopy. Organics and non-ionic components are usually more of a problem in water than salts are. I've always have more troubles using deionized water than I have had using distilled. And since distillation may not remove all ions, I've even had to use deionized-distilled.
I don't think water use is the issue here, though. Distillation does consume some water, but it shouldn't be wasting water. It does use more power. Deionization typically uses cartridges, which must be either regenerated (using water) or thrown away. Not counting the manufacture of the resins for the cartridges, or the salts added to your waste water stream. (And deionization consumes water also.)
So I would think distillation is the "better for the environment" solution, as well as being better for your solutions' environment.
Phil
**************************** =46rom: Donald Lovett {lovett-at-tcnj.edu}
My experience has been that from time to time, resin exchange deionized water results in random precipitates on sections. Switching to glass distilled water resolved the problem. I would use ion exchange water with caution.
*************************************** =46rom: DAVID PATTON {D-PATTON-at-wpg.uwe.ac.uk}
I have read about cases when EM stains have been adversely affected by contaminated (from filter breakdown products) de-ionised water. Mind you I have also read of stills with elevated Si in the water from the still glassware.
Dave **************************** =46rom: Glen MacDonald {glenmac-at-u.washington.edu}
We moved from a lab with deionized water feeding a glass still into a new facility with a reverse osmosis purifier followed by deonization/charcoal tanks, plumbed through the labs with a recirculating system. For EM stains, tissue culture, and molecular biology, we use this water after passing it through a Barnstead NANOpure polishing unit. The RO water is available in all labs and is sufficient for general histology and immunocytochemistry, and EM fixatives. The polishing unit is relatively conveniently located in our tissue culture lab. We have had no problems from the polished water in the 2 years that we have been using it.
Water distillers are no longer allowed to be installed at this campus due to their high energy costs and water inefficiency. My past experiences with non-distilled water systems left me very skeptical of deionization units until we installed our DI/destiller system in 1986. The still was partly to provide backup when the DI system failed. But, the DI system never failed. Our current system has been in use 2 years, so far the only problems have stemmed from lowest bidder installation, not from water quality.
********************************* =46rom: Robert Mixon {mixonr-at-ohsu.edu}
There are stills on the market which far exceed deionized water units in wat= er quality that not only re-use the cooling water in the distillation chamber (thus "preheated") but are also self-cleaning. True there is some "waste" of wate= r (could be saved as tap water to wash dishes I suppose??) I have no financial interest in any companies which produce such stills but have used one made b= y Weiss Scientific Glass of Beaverton, Oregon which at the time was the only o= ne on the market which recycled the water AND was self-cleaning (water exceeded some of the older "triple-distilled" models in quality). I am unrelated to and have no financial interest in the above mentioned company. Bob Mixon
We have had a couple of experiences with distilled vs deionised water issues. First when using glass distilled water our isolations of sperm cells from the pollen of Plumbago worked just fine. The university went to a campus wide Reverse Osmosis (RO) system and our isolations began to fail. Now we use RO water that has been deionised the isolations again work just fine. We are using a single high-efficiency DI filter cartridge which delivers water in the 18 MgOhm range according to the meter on the unit. Another related issue concerns the plumbing for the distilled/RO water supplied to the building. The taps that were installed were made of PVC which in itself is fine, but the anti-siphon valves that were installed were metal and incorrect for a distilled/RO water tap. We were informed that the lines should have had anti-siphon valves for DI/RO water installed and that the anti-siphon valves we had on there were probably leaching elements into the DI/RO water. The replacement taps (several of ours were broken) were of the metal variety and I was informed that they were tin lined and very expensive (thankfully replaced under repair to building!). So to make a long story short, we have had no troubles since going to DI water although we are deionizing water that has already been through reverse osmosis. For some reason the reverse osmosis water alone is not good enough for our purposes. As a secondary concern the way that the water is delivered may be an issue. I do not know if the anti-siphon valves were a problem or not as our isolations worked fine even though we had anti-siphon valves in place that were not "proper" for DI/RO water taps. As far as EM solutions - we use deionized water and have seen no problems. *************************************** End of messages
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
Hi! Anybody know anything about UNICO light microscopes? There are from United Products & Instrunents Inc. Any help or leads would be greatly appreciated. Feel free to contact me off-list at e-mail cited below. Thak yours, very much.
Gabriel Adriano Rosa Area Microscopia Electronica, Depto. Cs. Biologicas Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA
I try to measure fluorescence on non-transparent materials (e.g. plastics). What I need is a microscope or other scanning instrument with a resolution of about 1 =B5m. =46luorescence must be counted. The instrument must be very sensitive, the more, the better. (I suggest, laser as source for excitation should be the best). What is the most sensitive detection method? CCD and/or photomultiplier? Are there any systems commercially available? Thanks for answering or good tips! Armin Misch
In my past years as a microscope optical engineer, I tend to agree with = Russell. The fungi always attacked the soft internal lens coatings, not = the hardened coatings of exposed lens. If there is a problem on the = outside of the objective, I would first think of scratch damage or = similar. Whilst the fungi can be successfully removed from the lens = elements, it also means removing the soft coatings which in turn = compromise the chromatic correction of the lens. The coating can be = re-applied, however it is expensive. It is a popular practice for many = manufacturers of sea going binoculars to charge the body with nitrogen = and I know that at least one microscope manufacturer did this with = special order scopes to countries at risk. Also tried was an additive = to the coating material which, again available on special order, seemed = to increase the life of the optics.
The tips on prevention already discussed do go a long way in minimising = the risks. Some coatings seem to be better in this regard than others. = Also, to complicate matters more, there are variations in the quality of = the coatings from scope to scope.
Roger
Roger Wallis General Manager Optiscan P/L Confocal Microscopy PO Box 1066 Mt. Waverley MDC Victoria 3149 Australia Tel: (61) 3-9562-7741 Fax: (61) 3-9562-7742 e-mail: rogerw-at-optiscan.com.au URL: http://www.optiscan.com.au ______________________________
----------
One of the fungi that does this is Aureobasidium pullulans, it is=20 also seen often growing on tile grout in bathrooms. My best guess=20 it is using carbon in the coatings or cements, or thin oil films that=20 form from the air.
Russ
} Subject: Re: Lens-eating Fungi?? } Date sent: Wed, 25 Feb 98 09:47:11 -0400 } From: Dan {dan-at-bioptechs.com} } To: {Microscopy-at-Sparc5.Microscopy.Com}
} = ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of = America=20 } To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com } = -----------------------------------------------------------------------. } =20 } Credits to all that have contributed to this thread; } =20 } But does anyone know the name of this type of fungus? } What type of glass does it grow on and what component of the glass is = so=20 } attractive to it? } =20 } Before my business became overwhelmingly active I used to restore old=20 } optical systems for historical exhibition. } I no longer have time for that now but after years of seeing great = optics=20 } ruined by this fungus I am glad others have raised this topic. =20 } Hopefully I'll learn more about it. } =20 } Dan } =20 } =20 } =20 } =20 } Russell N. Spear Sr. Research Specialist Dept. of Plant Pathology Univ. of Wisconsin-Madison
Precise information about the goniometer tilt direction is important for many TEM applications. Historically alpha-MnO2 crystals have been used to measure the rotation angle between images and diffraction patterns in TEMs. However, for stereo analysis one needs to know the absolute tilt axis direction relative to the micrographs. I have used Kikuchi line motion to get this information. I am wondering what methods are used by others.
Salzburg, 26th of Febr., 1998, local time: 07.00 a.m.
Dear List-members, GOOD MORNING ALL
APOLOGIES and EXCUSES (or "NO EXCUSES ANY MORE" 0;) ):
} } For YOUR INFORMATION: { { THERE WILL BE NO POSTINGS ANY MORE, ESPECIALLY NO POSTING AS AN = "ATTACHMENT".
SIXTH INTERNATIONAL CONFERENCE AND WORKSHOP ON MOLECULAR MORPHOLOGY
to be held at: SALZBURG, Austria (Europe), OCTOBER 5-9, 1998
ORGANIZING FACULTIES: The University of Salzburg, Medical Research Coordination Center Salzburg General Hospital (St. Johanns-Spital LKA),=20 Institute of Pathological Anatomy The International Society of Molecular Morphology, Oklahoma City, OK, = USA The Cleveland Clinic Foundation, Cleveland, OH, USA
For FURTHER INFORMATIONS and for your convenience an
INTERNET-WEBPAGE is available (completed by March, 1st, 1998) at:
} } } } http://www.kongress.at/IMMC { { { {
For further informations, pre-registration and seat-reservation in = special workshops, please contact:
Prof. Dr. Gerhard W. HACKER Medical Research Coordination Center LKA SALZBURG c/o Dept. Pathol.-Anatomy Muellner Hauptstrasse 48 A-5020 SALZBURG, AUSTRIA Phone: ++43++662-4482-4730 ext., Fax: ++43++662-4482-882 ext., or:
e-mail: g.hacker-at-lkasbg.gv.at
Thank you very much for your attention and my } } deepest apologies { { to = you all reminding me of "NO ATTCHMENTS" to the LIST (I had a bad night = for those statements and reminders)
best regards to all of you, wish you a bright day
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
NO ANNOUNCEMENT of the Conference BY "ATTACHMENT(S)"
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YES, Codonics models 1600 (dye-sub) and 1660 (dye sub and thermal) are built around Kodak engines. In fact they physically look like the Kodak printers. However, the programming and other features are very different. The Codonics printers are UNIX machines complete with 540mb hard drive, floppy drive for updating programming and many additional features built into the software to permit much greater flexibility than the Kodak printer. They have a good reputation for mechanical reliability and Codonics has been quite good as far as phone support for questions, etc. We purchased a Codonics 1660 a few months ago and, although I have certainly made suggestions to the company to improve some features, overall we are quite pleased.
Debby Sherman, Manager Microscopy Center in Agriculture Purdue University
--------------------------------------
I would like to thank all of you for your responses about photo-quality printers. I will post a summary of responses in a day or two.
We have heard a rumor that the Codonics dye-sub printer is powered by the Kodak printer engine. Can anyone comment on this?
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Job opportunity: Automatic Diatom Identification and Classification (ADIAC)
A three-year fixed term position is available immediately at the Royal Botanic Garden Edinburgh as part of an EC-funded project (ADIAC) to create a system for identifying diatoms automatically by computer. The post will have a strong taxonomic component and will involve using existing database-management and image-analysis software to build a computerized database of names and digital images of diatoms which will be used as the essential foundation and reference for programs developed by a multinational team.
Applicants should have a PhD or equivalent, and relevant expertise, which could include one or more of the following: plant taxonomy, especially of algae or diatoms, light microscopy, digital image capture and manipulation, and familiarity with a wide range of computing techniques. Ability to meet deadlines and work in a team is essential.
In accordance with UK immigration requirements, priority will be given to EU nationals. Starting salary will be in the range GBP15,000-17,000, and will be pensionable. Applicants should send full cv including the names and addresses of two referees to the Personnel Department, Royal Botanic Garden Edinburgh, Edinburgh EH3 5LR, UK. Closing date 20 March 1998. Contact Stephen Droop at this address for further details, or e-mail: s.droop-at-rbge.org.uk.
*********************************************************** Stephen J.M. Droop Royal Botanic Garden, Edinburgh EH3 5LR, UK
We need to know what wavelength region you are interested in to answer your questions.
Laser(s) provide the brightest sources for your work, are "easiest" to couple into the microscope and ease the detection system requirements (less filtering problems, etc.). However, most provide only one wavelength. Multiple lasers may be employed but this can get expensive. Also, laser(s) are more likely to cause bleaching of the samples (or contaminants) that you might find confusing (or worse than confusing).
If you need to measure fluorescence over a broad spectrum, you may want to use an arc lamp and a number a matching spectral filters (Omega Optical makes a nice selection and has a good web site to explore). This may require you to make longer exposures.
CCDs can be more sensitive than PMTs in certain regions of the spectrum. A liquid nitrogen (LN)cooled CCD camera can be very sensitive and might be more useful for other experiments than PMTs.
At SEQ, we use both arc lamps and lasers. We also have used PMTs, LN cooled CCD cameras and image intensified cameras (for video rate microscopy).
P.S. Be very careful in cleaning and handling your plastics. Surface contamination could drive you crazy. Also, be sure you know the transmission/adsorption/reflection properties of pour plastic samples. This is particularly important if you are interested in a broad spectrum. In particular, many plastics can be quite transparent in the near infrared.
-----Original Message-----
Colleagues:
Any suggestions for this individual? Reply both to him and the listserver. He is not a subscriber.
I try to measure fluorescence on non-transparent materials (e.g. plastics). What I need is a microscope or other scanning instrument with a resolution of about 1 µm. Fluorescence must be counted. The instrument must be very sensitive, the more, the better. (I suggest, laser as source for excitation should be the best). What is the most sensitive detection method? CCD and/or photomultiplier? Are there any systems commercially available? Thanks for answering or good tips! Armin Misch
This inquiry pertains to the life of OSRAM HBO 103W/2 bulbs used for fluorescence microscopy.
During the last two years, and as recently as this morning, I've had several bulbs fail to light at low hour counts, e.g., 50-60 hrs. The problem does not seem to be the power supply, at least directly. New bulbs always light without fail. This causes me to think the problem involves a defect(s) in the bulb itself, perhaps induced by the power supply.
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2/26/98 9:39 AM Two-beam simulations
Jordi Arbiol i Cobos inquired
} Precise information about any computer program that allow me to } simulate the TWO -BEAM CONDITION. } Any help will be greatly apreciated.
We are currently running two-beam code for dislocation loop simulations.
One good source of information is the Oak Ridge National Laboratory report "Catalog of Computer Simulated TEM Images of Small FCC and BCC Dislocation Loops", by L. Sykes, W. Copper, and J. Hren, published February 1981. I believe the report number is ORNL/TM-7619. It includes a detailed discussion of two-beam dynamical simulation of dislocation loops and the report also contains a hard copy of their 2-beam simulation code. If you have any questions, feel free to contact me.
Tyrone L. Daulton Materials Science Division Argonne National Laboratory 9700 South Cass Ave Argonne, IL 60439
Irradiation Effects Group & Electron Microscopy Center Email: tyrone_daulton-at-qmgate.anl.gov Voice: 630-252-5079 Fax: 630-252-4798
Several months ago, I saw a small brochure on chromosome painting. I believe it was for mouse. I'm trying to remember the company that developed these kits and am having no luck. Does anyone out there have any experience with this?
I think we spoke on the phone about this once, because I was having the same problem with an old Zeiss Standard scope. Only shows up with video, not Polaroid, etc. Also MUCH more noticeable with low mag's (10x and less) than higher.
The only thing I found to help was to change the condenser lens. (I also tried diffusing near the lamp, having the scope completely cleaned and serviced, trying different objectives, etc.) If you're lucky you have a "flip-out" lever to remove the high mag. condenser when not needed (or desired) for the low mag objectives. If not, as my case, I had to unscrew the high mag. lens and remove it, but remind people who use higher mag's that they should put it back in when they use the scope. Also, trying a lower N.A. condenser might help. Also the Kohler illumination setup must be re-done after a change of condenser.
Good luck, Karen
dkittleson-at-pillsbury.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have an early 1980's vintage Zeiss Universal pol. scope with a } Diagnostics Instruments adapter and Sony 960 CCD camera. There is an } annoying "hotspot" visible in both the live and stored images. This is } accentuated in certain instances such as low contrast, slightly crossed } polars, etc. The hotspot is almost undetectable in images of samples } diffusing a significant amount of light. It is my opinion that the } microscope optics are contributing this problem. However, I certainly } have an open mind. I have made the following observations: } -The microscope is set-up for Kohler illumination. Inserting the } diffuser before the lamp housing produces only a slight improvement. } -There is virtually no issue when using this camera with the appropriate } adapter for our Wild M400 photomacroscope or Olympus Provis Light } microscope. } -I have tried 2 different Diagnostic Instruments adapters (.6x, .45x) and } 3 sets of adapter lenses. In addition, a Dage camera (c-mount) to a } "live" monitor shows the same hot spot. } -Pol. objectives of that vintage were not flat field. I tried a series } of flat field objectives...alas...the field is flat...the hot spot } remains. } -Local service reps have not been able to implicate the source. } -Polaroid and 35mm imaging produces no visible "hot spot". } I would be extremely grateful to anyone who could offer suggestions as I } have agonized over this for quite some time. Thank you. } } Diana Kittleson } Pillsbury Technology East } 737 Pelham Blvd. } St. Paul, MN 55114 } dkittleson-at- pillsbury.com
-- Karen Zaruba, Life Sciences Sector Laboratory, 3M Company, St. Paul, MN 55144 kszaruba-at-mmm.com
"If you can spend a perfectly useless afternoon in a perfectly useless manner, you have learned how to live." - Lin Yu Tang [Of course, a perfectly useless afternoon is a rare thing indeed!] *The opinions above are my own, not necessarily my employer's*
Rick Felten 02/26/98 01:25 PM I would like some opinions about the price/performance ratios of upgrading my EDS system through IXRF vs Kexex. I am currently using a Hitachi S2400 SEM. I am interested in thoughts about the maturity of the software, level of "Bugs", and ability to provide both hardware and software service. Please include your experience with either.
Thanks for your comments
Ric Felten of MacDermid Inc., a Specialty Chemical Company in Waterbury, Ct USA
I have not been paying attention to this thread, so don't know if it has been pointed out that SINCE this problem is with video only, the gain/contrast setting is probably contributing. This is an adjustment which is not available with film cameras. I have a similar experience with my nice Olympus BX-50 when I boost the contrast tremendously to see objects with little inherent contrast. I am clearly seeing spatial variation in illumination, which I can't eliminate by adjusting the lamp or adding a mild diffuser. CCD cameras have such a wide range of adjustment that they can easily show the limitations in design of even pretty good pretty recent scopes.
Of course there will be more spatial variation at low mag than at hi mag. For a perfectionist using hi mag only, the solution might be a Fiber optic light scrambler, though this may not work at low mag. see http://www.technicalvideo.com/. Otherwise, check your illumination & optimize it as much as possible.
I think we spoke on the phone about this once, because I was having the same problem with an old Zeiss Standard scope. Only shows up with video, not Polaroid, etc. Also MUCH more noticeable with low mag's (10x and less) than higher. . . . } -----------------------------------------------------------------------. } } I have an early 1980's vintage Zeiss Universal pol. scope with a } Diagnostics Instruments adapter and Sony 960 CCD camera. There is an } annoying "hotspot" visible in both the live and stored images. This is } accentuated in certain instances such as low contrast, slightly crossed } polars, etc. The hotspot is almost undetectable in images of samples } diffusing a significant amount of light. It is my opinion that the } microscope optics are contributing this problem. . . . . . . Karen Zaruba,
"If you can spend a perfectly useless afternoon in a perfectly useless manner, you have learned how to live." - Lin Yu Tang [Of course, a perfectly useless afternoon is a rare thing indeed!] *The opinions above are my own, not necessarily my employer's*
This is a multi-part message in MIME format. --------------A92459C6DB2E52D547FE1B7E Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit
Diana: I had a similar problem with our Zeiss Standard scope when we worked with low mag obj. lenses using a video capture camera in the third ocular tube. The problem was solved by painting the front reflective surface of the metal adapter on the end of the camera with carbon dag. Apparently light reflected from this surface was causing the problem. Doug Bray
I think we spoke on the phone about this once, because I was having the same problem with an old Zeiss Standard scope. Only shows up with video, not Polaroid, etc. Also MUCH more noticeable with low mag's (10x and less) than higher.
The only thing I found to help was to change the condenser lens. (I also tried diffusing near the lamp, having the scope completely cleaned and serviced, trying different objectives, etc.) If you're lucky you have a "flip-out" lever to remove the high mag. condenser when not needed (or desired) for the low mag objectives. If not, as my case, I had to unscrew the high mag. lens and remove it, but remind people who use higher mag's that they should put it back in when they use the scope. Also, trying a lower N.A. condenser might help. Also the Kohler illumination setup must be re-done after a change of condenser.
Good luck, Karen
dkittleson-at-pillsbury.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have an early 1980's vintage Zeiss Universal pol. scope with a } Diagnostics Instruments adapter and Sony 960 CCD camera. There is an } annoying "hotspot" visible in both the live and stored images. This is } accentuated in certain instances such as low contrast, slightly crossed } polars, etc. The hotspot is almost undetectable in images of samples } diffusing a significant amount of light. It is my opinion that the } microscope optics are contributing this problem. However, I certainly } have an open mind. I have made the following observations: } -The microscope is set-up for Kohler illumination. Inserting the } diffuser before the lamp housing produces only a slight improvement. } -There is virtually no issue when using this camera with the appropriate } adapter for our Wild M400 photomacroscope or Olympus Provis Light } microscope. } -I have tried 2 different Diagnostic Instruments adapters (.6x, .45x) and } 3 sets of adapter lenses. In addition, a Dage camera (c-mount) to a } "live" monitor shows the same hot spot. } -Pol. objectives of that vintage were not flat field. I tried a series } of flat field objectives...alas...the field is flat...the hot spot } remains. } -Local service reps have not been able to implicate the source. } -Polaroid and 35mm imaging produces no visible "hot spot". } I would be extremely grateful to anyone who could offer suggestions as I } have agonized over this for quite some time. Thank you. } } Diana Kittleson } Pillsbury Technology East } 737 Pelham Blvd. } St. Paul, MN 55114 } dkittleson-at- pillsbury.com
-- Karen Zaruba, Life Sciences Sector Laboratory, 3M Company, St. Paul, MN 55144 kszaruba-at-mmm.com
"If you can spend a perfectly useless afternoon in a perfectly useless manner, you have learned how to live." - Lin Yu Tang [Of course, a perfectly useless afternoon is a rare thing indeed!] *The opinions above are my own, not necessarily my employer's* -------------------------------------------------------------------------------- Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: from Sparc5.Microscopy.Com by admin3.cc.uleth.ca (MX V4.2 AXP) with SMTP; Thu, 26 Feb 1998 12:28:17 MDT Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id LAA28261 for dist-Microscopy; Thu, 26 Feb 1998 11:43:46 -0600 Received: from sulu.mmm.com (sulu.mmm.com [192.28.4.21]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id LAA28258 for {microscopy-at-sparc5.microscopy.com} ; Thu, 26 Feb 1998 11:43:45 -0600 Received: (from daemon-at-localhost) by sulu.mmm.com (8.8.7/8.8.1) id LAA29332 for microscopy-at-sparc5.microscopy.com..; Thu, 26 Feb 1998 11:46:17 -0600 (CST) Message-ID: {34F5B27F.153A-at-mmm.com}
At 11:08 AM 2/26/98 -0800, you wrote: } } I have not been paying attention to this thread, so don't know if it has } been pointed out that SINCE this problem is with video only, the } gain/contrast setting is probably contributing. This is an adjustment which } is not available with film cameras. I have a similar experience with my } nice Olympus BX-50 when I boost the contrast tremendously to see } objects with little inherent contrast. I am clearly seeing spatial variation in } illumination, which I can't eliminate by adjusting the lamp or adding a mild } diffuser. CCD cameras have such a wide range of adjustment that they } can easily show the limitations in design of even pretty good pretty recent } scopes.
Interesting thought that the camera gain may simply be pointing out what has been there all along. I wonder if digitizing/scanning a photo and taking the image to a suitable program for contrast enhancement and a line profile would show that the illumination is uneven, but just not apparent on normal photos. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
Hank- I think you have a great idea, but you'll probably have to follow up on it yourself. Try to keep it all confidential, and use some sort of criteria such as, mat sci or biol, industry vs academia, experience (1-5 yrs, 5-10 yrs, over 20, etc.), also throw in some regional and or cost of living factor. I'll be interested in your findings. -Mike
James-be careful, don't get shocked check the terminals at the positive & negative ends of the bulb, are they secure? next check the wires to the terminals, trace them to their source, then keep tracing ... step by step, to the power supply (I don't suggest opening the power supply, unless you feel confident in what you're doing), be careful, don't get shocked. -Mike
On Thu, 26 Feb 1998, James Martin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } This inquiry pertains to the life of OSRAM HBO 103W/2 bulbs used for } fluorescence microscopy. } } During the last two years, and as recently as this morning, I've had } several bulbs fail to light at low hour counts, e.g., 50-60 hrs. The } problem does not seem to be the power supply, at least directly. New } bulbs always light without fail. This causes me to think the problem } involves a defect(s) in the bulb itself, perhaps induced by the power } supply. } } Any thoughts? } } James Martin } Williamstown Art Conservation Center } } }
I have receieved the following email from a friend in New Zealand can anybody help - thanks. Steve
Precise information about the goniometer tilt direction is important for many TEM applications. Historically alpha-MnO2 crystals have been used to measure the rotation angle between images and diffraction patterns in TEMs. However, for stereo analysis of nano crystals on high resolution TEM images one needs to know the absolute tilt axis direction relative to the micrographs. I have used Kikuchi line motion to get the tilt axis direction in diffraction patterns. Then the rotation angle between diffraction patterns and images enables the tilt axis in high resolution TEM images to be calculated. I am wondering what methods are used by others.
Background subtraction would seem to be appropriate.
Richard
At 11:08 AM 2/26/98 -0800, you wrote:
Interesting thought that the camera gain may simply be pointing out what has been there all along. I wonder if digitizing/scanning a photo and taking the image to a suitable program for contrast enhancement and a line profile would show that the illumination is uneven, but just not apparent on normal photos. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
Readers, While we would all agree that microscopy is "serious science", to have a bit of fun when possible might not be too bad an idea. With this though in mind, I would like to announce a "Composite Image Contest" to be held out of our booth at the upcoming MSA/MAS Conference in Atlanta (July 12/16 1998). - Entries must be a composite of two or more images, one at least must be microscopical in nature. They must be in hard copy (photographic or printer output) and while OK in any size, ~ 8.5" x 11" is preferred. - To be clear, we are looking for "fun" not technical stuff. Like, how about Clinton's face on an insects body, or how about a partially nude female backstroking in a "sea" of mosquito larva? Whatever! - Conference attendees will vote on the entries based on how interesting and creative. First prize will be a 1 oz Maple Leaf gold coin (~$450 value), 2nd and 3rd prizes to be announceed. All contestants will receive their choice of two microscopical prints by David Scharf. - Entries are welcome not only from U.S. microscopists, but from "overseas" microscopists as well as manufacturers and suppliers. You will not have to be present to win. Contestants may submit a maximum of two entries. - Winning images will be featured on the cover of "Microscopy Today" IMPORTANT - this contest is not to be confused with the Micrograph Competition. - If you think that you may (repeat, may) be interested in participating, kindly advise direct to me. I will then keep you advised of any information regarding the contest. I sure hope for your interest! Don Grimes, Microscopy Today
Dear all, We have an antique Hitachi s410 that used to function quite well. Recently it from time to time came out images with zig zag pattern. We checked with local service engineers a couple of time but not able to figure out what the real cause is. Based on the engineer's dignose, we seem suddenly have some magnetic field interference from somewhere. Since we are going to install a new one pretty soon. This magnetic thing seems will be a big pain in the future too. What I would like to know is anyone outthere have similar experience can share
Dear Kuo, There may be several possible causes for this - physical vibration, electronic noise in the deflection electronics, or even SED arcing. I would not suspect magnetic fields as the culprit unless the mu metal shielding has been removed. It is difficult to diagnose the problem without seeing the image. Maybe you could email a tiff of what you are seeing to my address. AZ
Andrew L. Zobel Service Engineer ________________________ RJ Lee Instruments Ltd. http://www.rjleeinst.com
Dear all, We have an antique Hitachi s410 that used to function quite well. Recently it from time to time came out images with zig zag pattern. We checked with local service engineers a couple of time but not able to figure out what the real cause is. Based on the engineer's dignose, we seem suddenly have some magnetic field interference from somewhere. Since we are going to install a new one pretty soon. This magnetic thing seems will be a big pain in the future too. What I would like to know is anyone out there have similar experience can share. Will this image pattern could possible caused by aging scope per se ? Also, how could I can really measure the magnetic field in my lab and locate the disturbance? Thanks ahead! KerChung Kuo
I have this similar problem on a Reichart, it is only observed at lower magnifications. Without success, I have adjusted the bulb, illuminators, everything, including disassembly of the apertures. I also thought it might be the optics in my video adapter, but it sounds like others interchange video and Polaroid, which excludes this as a source.
My current recourse is use use an image processing filter (from John Russ' toolkit) with Photoshop to minimize illumination variables. This usually produces excellent results, but occassionly produces some "exotic and colorful" artifacts.
I am no expert in this subject but field measurements can be made by placing a toroidal coil (with a known number of turns) at various locations in the room (3ft high and every 5ft for instance) and measuring the induced voltage at each location. Through Faraday's law (I think), the flux in webers can be calculated and plotted for the room. Hope this helps, AZ
Andrew L. Zobel Service Engineer ________________________ RJ Lee Instruments Ltd. http://www.rjleeinst.com
James How often do you switch on, and how long do you keep the bulbs burning once lit? I understand that the main wear on HBOs is the number of times switched on, more so than the number of hours lit.
Stephan Helfer Royal Botanic Garden Inverleith Row Edinburgh EH3 5LR Scotland UK
} Precise information about any computer program that allow me to } simulate the TWO -BEAM CONDITION. } Any help will be greatly apreciated.
The following announcement was sent to the discussion group a couple of years ago. I have not used the program but retained the message in case of needing this program or similar at some point in the future. I would guess that it is based on the original Fortran code by Head, Humble et al. (early 1970's as I remember) but is probably a more modern implemention of this code.
Hope this helps
Ian
} Date: Thu, 25 Jul 1996 13:26:00 +0200 } From: Koenraad.Janssens-at-mtm.kuleuven.ac.be (Koenraad Janssens) } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: SIMCON } X-Sun-Charset: US-ASCII } } Dear Fellow Microscopists, } } The last few years I have been involved in research on localized (near } nanometer resolution) strain characterization using two-beam electron } diffraction contrast imaging (an operational mode of transmission } electron microscopy). See any book on transmission electron microscopy } for the basics. } } Part of this project was to develop a software package allowing for the } analysis of strain fields of arbitrary geometry. This software (SIMCON) } allows one to model a microscopic strain field with various mathematical } methods (including finite elements) and subsequently verify this model } by comparison with experimental observations in the TEM. } } As of today we are releasing SIMCON as freeware, you can find out all } details at the following address: } } http://www.mtm.kuleuven.ac.be/~simcon/ } } Friendly Greetings, } } Koen Janssens } } ___________________________________________________________________ _ _ _ } ___________________________________________________________________ _ _ _ } } Koenraad Janssens, Ph.D. } } KULeuven } Department of Metallurgy and Materials Engineering (MTM) } de Croylaan 2, B-3001 Leuven, Belgium } Tel. : +32-(0)16-32.1232 } Fax : +32-(0)16-32.1992 } e-mail : Koenraad.Janssens-at-mtm.kuleuven.ac.be } www : http://www.mtm.kuleuven.ac.be/Members/Researchers/KoenraadJanssens/ } } } !!! New address valid from 1 September 1996 !!! } } OCAS } John F. Kennedylaan 3, B-9060 Zelzate, Belgium } Tel. : +32-9-345.12.11 (OCAS reception) } Fax. : +32-9-345.12.04 }
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
You are quite correct, the problem with this is making the coil just right and then reading the waveforms you get. You need to understand the waveforms to get a value for the interference. Even on the calibrated coil we have, it is difficult reading the oscilloscope waveforms and giving the customer a value in milli gauss.
Luc Harmsen Anaspec, South Africa Technical support for E.M. operators, world wide. anaspec-at-icon.co.za TEL: ++ 27 (0) 11 476 3455 FAX: ++ 27 (0) 11 476 7290
-----Original Message-----
I am no expert in this subject but field measurements can be made by placing a toroidal coil (with a known number of turns) at various locations in the room (3ft high and every 5ft for instance) and measuring the induced voltage at each location. Through Faraday's law (I think), the flux in webers can be calculated and plotted for the room. Hope this helps, AZ
Andrew L. Zobel Service Engineer ________________________ RJ Lee Instruments Ltd. http://www.rjleeinst.com
FYI Many people have experienced this hot spot problem with tv images because they used the wrong or inferior tv adapter. Make sure that you are using the correct coupler for your size chip or tube camera. I suggest you contact your Carl Zeiss rep in your area, 1-800-543-1033 Just my .02 worth. J Reber Carl Zeiss
We also had a problem with bright spots in our digital image, using a Polyvar microscope. As was mentioned by one respondent, the cause was internal reflections from the adapter between the microscope port and the lens of the camera. We used carbon dag, camera black and a couple of other coatings but still suffered some internal reflections. We finally used a piece of flat black construction paper to line the inside of the adapter tube. This works in most cases but there are times when we use neutral density filters to reduce the amount of light going to the camera. Hope this helps.
David O'Neil tel: (902) 426-8258 National Research Council of Canada fax: (902) 426-9413 Institute for Marine Biosciences 1411 Oxford St. Halifax, Nova Scotia B3H 3Z1 Canada email: david.o'neil-at-nrc.ca
We also had a problem with bright spots in our digital image, using a Polyvar microscope. As was mentioned by one respondent, the cause was internal reflections from the adapter between the microscope port and the lens of the camera. We used carbon dag, camera black and a couple of other coatings but still suffered some internal reflections. We finally used a piece of flat black construction paper to line the inside of the adapter tube. This works in most cases but there are times when we use neutral density filters to reduce the amount of light going to the camera. Hope this helps.
David O'Neil tel: (902) 426-8258 National Research Council of Canada fax: (902) 426-9413 Institute for Marine Biosciences 1411 Oxford St. Halifax, Nova Scotia B3H 3Z1 Canada email: david.o'neil-at-nrc.ca
I have also had this problem with HBO 103W/2 bulbs, a talk with our vendor revealed this is a common problem. We solved it by changing to HBO 100W/2's.
Russ
} Date sent: Thu, 26 Feb 1998 10:36:36 -0500 (EST) } From: James Martin {James.S.Martin-at-williams.edu} } Subject: OSRAM HBO 103W/2 bulbs } To: microscopy-at-Sparc5.Microscopy.Com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } This inquiry pertains to the life of OSRAM HBO 103W/2 bulbs used for } fluorescence microscopy. } } During the last two years, and as recently as this morning, I've had } several bulbs fail to light at low hour counts, e.g., 50-60 hrs. The } problem does not seem to be the power supply, at least directly. New } bulbs always light without fail. This causes me to think the problem } involves a defect(s) in the bulb itself, perhaps induced by the power } supply. } } Any thoughts? } } James Martin } Williamstown Art Conservation Center } } } Russell N. Spear Sr. Research Specialist Dept. of Plant Pathology Univ. of Wisconsin-Madison
PLease help me. I am trying to learn how to use a cryo-ultra microtome. How in the world do you get the ultra-thin sections on the grids? Also, how do you store them? How long can you keep them? ETC.(I am sure I don't even know what else to ask) Thank you, Sally Shrom
I have a question about the 180 degree ambiguity discussion. Doesn't the shadow image of an under-condensed BF disk of a convergent beam pattern determine the rotation angle of the diffraction pattern with respect to the image without the ambiguity? If it isn't, then I may have been assigning incorrect indices to some III-V XTEM samples that I've done.
Our current shipment of HBO103 bulbs seems to work fine. Maybe too well; we've had problems with the older Zeiss lamp housings getting burned out. But the Nikon housings and the new Olympus lamp housings are holding up ok. In the past we've had problems with the Osram HBO 103 bulbs. They became unstable after very little usage and fogged quickly. If you can get Ushio bulbs, or can switch to 102 instead of 103, this would probably be better.
-------------------------------------------- Michael Cammer email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 http://leper1.ca.aecom.yu.edu/aif/ --------------------------------------------
} Background subtraction would seem to be appropriate. } } Richard } } } At 11:08 AM 2/26/98 -0800, you wrote: } } Interesting thought that the camera gain may simply be pointing out what } has } been there all along. I wonder if digitizing/scanning a photo and taking the } image to a suitable program for contrast enhancement and a line profile } would show that the illumination is uneven, but just not apparent on } normal } photos. } ---------------------------------------------------- } Warren E. Straszheim } 23 Town Engineering } Iowa State University } Ames IA, 50011 } Phone: 515-294-8187 FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } http://www.marl.iastate.edu } } electron microscopy, x-ray analysis, image analysis, computer } applications } }
Rather than background subtraction, you may want to flatfield your image, i.e. divide by an image obtained on an empty field (that is with everything but your sample, such as an empty spot on a microscope slide, etc.). This will also correct for any nonuniformity of sensitivity in your camera and other optical artifacts leading to uneven sensitivity.
Massimo
_____________________________________________________ Massimo Sassaroli, D.Sc. Department of Physiology and Biophysics Box 1218 Mount Sinai School of Medicine One Gustave L. Levy Place New York, NY 10029-6574
This contest would be a lot more fun for me if I hadn't read the line about a "partially nude woman." Why, why, why was it even necessary to bring this element into an otherwise fun and entertaining exercise???? I'm fighting the urge to turn this into a full-blown diatribe, but for now I'll just say, I find this image offensive.
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064
Are we all talking about the same thing here? I wonder if there is a difference between the "hot spot" vs. "bright spots" that people are seeing. At least in my case, there were no "bright spots", per se, just a drastic gradient in brightness from the center of the field (hot spot) to the edges. Is this what the people with problems (reflections) in their camera adapters were seeing?
Regarding the tips about background subtraction, flattenting, etc. In fact that was what I had to do for a long time. But I am leery of doing this sort of manipulation if greyscale density values are to be collected. Is it considered acceptable?
Karen -- Karen Zaruba, Life Sciences Sector Laboratory, 3M Company, St. Paul, MN 55144 kszaruba-at-mmm.com
*The opinions above are my own, not necessarily my employer's*
Hi, Sally There are excellent references following:
Book: G.Griffiths. Fine structure immunocytochemistry. Springer -Verlag, New York, Berlin, Heidelberg
Papers: Liou W,Geuse HJ,Slot JW. Improving structural integrity of cryosections for immunogold labeling. Histochem Cell Biol(1996) 106: 41-58
Tokuyasu KT. J Cell Biol (1973) 57:551-565
Tokuyasu KT. J. Microsc. (1986) 143:139-149
Tokuyasu KT. Histochem J (1989) 21: 163-171
Best regards, Alexander A. Mironov Jr.
On Fri, 27 Feb 1998, Sally Shrom wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } PLease help me. I am trying to learn how to use a cryo-ultra microtome. } How in the world do you get the ultra-thin sections on the grids? Also, } how do you store them? How long can you keep them? ETC.(I am sure I don't } even know what else to ask) } Thank you, } Sally Shrom } }
Did I miss something? What is fun about an image of a partially nude woman swimming in a sea of larvae?
There is much that could be said about this, but suffice my own comments to be: rethink your offer, Mr. Editor, and try rephrasing in a more respectful manner.
Group - In recently announcing the "Just-For-Fun" contest, I suggested that a "partially nude" female swimming in lavra - and two of you were quite offended. Perhaps if I would have said "partially clothed" rather than partically nude, it would have been acceptable. As a young man I clearly recall Ester Williams swimming arouind (partically nude or partically clothed, depending on your view) in the movies - and it is tough for me to see how anyone today could be offended. However, since two of you obviously were, I sincerely apologize. I was only seeking some other kind of composite example rather than a face on a bugs body. Don Grimes, Microscopy Today
I suspect that Jane and others (myself included) were offended by the sexist nature of the suggestions for the "Just For Fun." announcement not the nudity although I'm sure some people are offended by nudity. We'll see if men, nude or partially clothed get equal time with women. I'll submit an appropriate image as an experiment. This will be fun!
Leica, Inc., Diatome US, and Electron Microscopy Sciences announce another in a series of "ultramicrotomy of materials" workshops. This seminar will focus on the hands on participation of the following technics:
Embedding of industrial samples Specimen trimming
Ultramicrotomy of hard materials Ultramicrotomy of polymers
Collection & handling of sections Staining of polymer sections
Low temperature ultramicrotomy SEM applications of ultramicrotomy
The format of our workshop is half day group lecture and half day lab work in small groups. Video attachments will be used on ultramicrotomes in order to maximize the learning experience. Participants are encouraged to bring their own samples to work with in addition to those supplied by the course instructors.
Course Speakers & Instructors:
Dr. Tom Malis Mrs. Ani Issaian, M.S. Canadian Federal Laboratory California Institute of Technology Characterization Group Leader Dept. of Chemical Engineering Researcher of polymer physics
Mr. Helmut Gnagi Ms. Kathy Johnson Product Manager Materials Analyst Diatome, Ltd. Switzerland Gates Rubber Co. Denver, CO
Where: University of Colorado High Voltage EM Lab Lab for 3-D of Fine Structure Dr. Richard McIntosh Mr. Mark Ladinsky
When: May 19-21, 1998
Tuition: $1,100.00 US includes three nights stay at the Regal Harvest House Hotel, continental breakfast, lunch, and dinner daily, course supplies, and lab charges.
unsubscribe Lic. Veronica Campanucci -------------------------------------------------------------------------------- Lic. Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
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This contest would be a lot more fun for me if I hadn't read the line about a "partially nude woman." Why, why, why was it even necessary to
bring this element into an otherwise fun and entertaining exercise???? I'm fighting the urge to turn this into a full-blown diatribe, but for now I'll just say, I find this image offensive.
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064
by dialer.laval.com (8.8.8/8.8.7) with SMTP id SAA12111; Fri, 27 Feb 1998 18:29:58 -0500 Message-Id: {3.0.3.32.19980227182657.00a4c508-at-laval.com} X-Sender: laurier-at-laval.com X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.3 (32)
Hi This is a transformer design problem. Not the bulb, the other solution would be to change the transformer. Norm At 08:25 27/02/98 CST, Russell Spear wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good retraction Don! and oh how the times have changed, it is tuff to be (PC) politically correct these days. good luck with the contest, it should be FUNN;-) -Mike PS- I'll be forwarding some info. for the salary survey, thanks for taking it on.
If anyone is going to be attending the PITTCON 98 in New Orleans, by all means don't miss the opening session on Sunday night because one of the invited speakers is our own Dr. Nestor Zaluzec. This is, incidentally quite a high honor to serve as one of the opening session speakers.
The opening session starts at 7:00 pm and you can get full details from the PITTCON website, specifically at URL http://www.pittcon.org/opening.htm
The topic of the opening session is "Exploring the Future of Science and Instrumentation on the WEB."
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
First, Nestor, I agree that enough is enough on this thread. However, my screw-up has opened up a topic that must be of interest to all of us. I have received dozens of comments on this topic - and resist a summary of the numbers in support of, against, etc. However, I submit that the following might be of interest to many:
It starts out as an Employee to University "X", it continues as: and all employees have been 'strongly encouraged' to attend a mini conference on sexual harassment, in recent years. Part of what I learned is that there is the potential for someone to take offense at almost anything of gender/sexual nature. Thus, in the workplace comments of that nature are best avoided all together. I know times have changed, and there are far more women in the workplace overall, and certainly more women working in professional capacities. As a woman I find the quality of workplace life is higher when I just do not have to encounter those 'sensitive ' topics. I also believe you did not intend to give offense, and hope that the contest is successful. Part of my personal experience with the changing attitudes about acceptable behavior and comments in the workplace include observing my father, who I regard as a sensitve and responsible man, respond to changing requirements in this area in the federal service. Even nice guys like him, who I have never seen treat a woman disrespectfully, have had to adapt to a certain degree.
Expecting that I am of the age of her father, it seems that I (and maybe you?) must be more careful in how we comment on life?
Let's try this again without the truncation of the message.
I have a question about the 180 degree ambiguity discussion. Doesn't the shadow image of an under-condensed BF disk of a convergent beam pattern determine the rotation angle of the diffraction pattern with respect to the image without the ambiguity? If it isn't, then I may have been assigning incorrect indices to some III-V XTEM samples that I've done.
I don't think I have ever met anyone who has not made a slip of the tongue, pen, computer, or what-have-you that they have regretted. Most times, these things are done in total innocence, no harm intended. One was certainly made here, but please remember that one on-line slip may have offended some folks, but many on-line angry rebuttals can damage or even destroy a career. We live in very sensitive times.
I do not condone sexism, but I also do know that even the best intentioned of us can mess up once in a while.
No real harm has been done to anyone, yet. Let's slow down and keep it that way. Please. I expect the point has already been made.
Best wishes, Randy
Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
I thought the idea was to have some fun. Come on ladies and lighten up!! No offense was meant and none should be taken. I hate it when everyone starts bickering about some trivial point which has really nothing to do with the original subject. I subscribe to this list serve in order to hear the usual intelligent exchange of advice and ideas on topics related to microscopy. The gentleman has proposed an entertaining contest for our entertainment and amusement. He playfully described an image which he thought would be an illustrative example of the type of imaging he was refering to. Nothing lewd or offensive was implied in so far as I can tell. The image of a woman in a bathing suit (with or without larvae) has never been offensive to me. Come on sisters, lighten up and have some fun already!
Now, can we get back to microscopy topics and not take off on another tangent!
Anne Marie Cooper
On Fri, 27 Feb 1998, Elinor Solit wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Did I miss something? What is fun about an image of a partially nude } woman swimming in a sea of larvae? } } There is much that could be said about this, but suffice my own comments } to be: rethink your offer, Mr. Editor, and try rephrasing in a more } respectful manner. } } Elinor Solit } The Microscope Book } }
The following info relates to cryo ultramicrotomy of polymers, which may, or may not help you with your current application. =20
After sectioning the sample, the sections are fished off the edge of = the diamond/glass knife by means of a thin wire loop about 2 mm. in = diameter which holds a drop of concentrated sucrose solution. The drop is touched to the sections before the solution freezes, and then touched = to a TEM grid. The sections and some of the solution are thus transferred to the grid which is then floated on distilled water to leach away the sucrose. Don=92t worry - the sections remain on the grid. You can = then pick up the grids with tweezers and lay them on filter paper to soak up the water.
I suggest that you get in touch with Prof. Karen Winey who is also at UPenn. and she can tell you more about the technique. Her phone number is 898-0593. Tell her I said Hi!
Ramon J. Albalak, D.Sc. Research and Development Directorate Carmel Olefins Ltd. POB 1468 Haifa 31014 Israel
---------- From: Sally Shrom[SMTP:sally-at-retina.anatomy.upenn.edu] Sent: Friday, February 27, 1998 16:33 To: Microscopy-at-Sparc5.Microscopy.Com Subject: cryo-thins TEM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20 To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com =09 -----------------------------------------------------------------------.=
PLease help me. I am trying to learn how to use a cryo-ultra microtome. How in the world do you get the ultra-thin sections on the grids? Also, how do you store them? How long can you keep them? ETC.(I am sure I don't even know what else to ask) Thank you, Sally Shrom =09
Our company can provide you with YAG:Ce screens in various thicknesses if you will please contact us directly.
For general information we offer the following : For thin screens obtain= ed from a thicker disc it is best to adhere the crystal to your window or fi= ber optic plate; after pre thinning to ~0.25 to 0.4mm thick. This gives a better chance of surviving without cracking. We have had windows polished= to 0.25mm in the past with a survival rate (i.e. no cracks) of 45%. I presu= me you must be backing the window up anyway if there is a vacuum differentia= l . The largest diameter single crystal readily available is about 35mm. Although 50mm is not out of ones growing capability, it would be rather expensive to process. Another problem notorious with single crystal YAG:C= e is the distribution of the Cerium dopant (undoped YAG will not work). As = one gets thinner this problem could be even more noticeable if areas were voi= d of the dopant due to decreased volume. One might argue it's only the top = few microns that really matter so if you had a 0.5mm thick crystal that gave = a good signal try polishing from the back side; thereby leaving the known = good surface unchanged. Of course that leaves you with the problem of how to adhere the "good" surface to a back up plate for grinding and polishing t= he back, and then removing it to remount the back against the window!! Your optical technician is sure to have some tricks.
We are currently in the process of prototyping ultra thin crystals grown = on quartz or sapphire substrates. In this case neither thickness or size/diameter would be very limited. These would not necessarily be singl= e crystal, but some trials in a TEM will verify image quality. If you have = an interest in this alternative approach please contact us. There are sever= al researchers going in that direction and I wouldn't be at all surprised if they may be already on the market, although not necessarily the EM market. The EM market is relatively small for this material currently compared to other markets (lasers for example) where other materials are = more lucrative for crystal growers. YAG:Ce is not something other markets have required to satisfy their need; therefore, it is rather expensive compar= ed to crystals of other types.
Finally old screens can be rejuvenated in some cases by cleaning and recoating; assuming they are not defective for a variety of other reasons= . Contamination on the surface will give the appearance of an otherwise perfectly good screen. This is particularly true if your system has any o= il pumps (roughing, backing, or diffusion ) or rubber o-rings to produce contamination. A replaceable element foreline trap will help to trap oil vapors and prolong the screen lifetime.
Hope this helps.
M.E. Taylor Engineering Inc. 21604 Gentry Lane Brookeville, MD 20833 Phone: 301-774-6246 =95 FAX: 301-774-6711 =95 e-mail: Metengr-at-aol.com
DTSA version 2.0.1 for the Macintosh (available from NIST) can open these files directly by using the menu item "Read sundry file formats" and choosing "TN 'XI' files." However, this menu item was removed from version 2.5 for the PowerPC. I don't know if the newer versions perform this function in a new way....but for now, we're keeping the old version of DTSA in the lab.
Jeffrey A. Fortner Nuclear Waste Management Section Chemical Technology Division Argonne National Laboratory Argonne, IL 60439-4837
} ---------- } From: gllovel-at-ppco.com } Sent: Friday, February 20, 1998 11:05 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Spectra Xfer#2 } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Since my last posting about transfering TN-5500 spectra to a PC I have } managed to send spectra information( headers and each channels } intensity) as } ASCI data to the PC. I have not however found a means to convert the } data } into a graphics file. Kermit was used to capture the xfered data. } The PC } is connected to a Novel network but the only means of TN-5500 data or } spectra to be xfered is by means of a serial port connection beween } the two } systems. Any information concerning software that can be located on } the PC } for converting the data to a graphics file and then converting the } graphics } file to a TIF file would be greatly appreciated. If anyone is } interested, } the spectra data was xfered by using Noran's XI module(file type 5, } number } 442), which requires two other type 6 files( 4000 and 4065 ) be } present on } the operating system. }
You may try CUFOUR for a two-beam calculation. There is an on-line limited version running on http://cimewww.epfl.ch/CIOL/html/ems.html If you are interested in acquiring CUFOUR or need more information let me know!
Yours,
Robin Schaublin
arbiol-at-iris1.fae.ub.es wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Precise information about any computer program that allow me to } simulate the TWO -BEAM CONDITION. } Any help will be greatly apreciated. } } Thank you very much for your time. } } Yours respectfully, } } Jordi Arbiol i Cobos } Departament d'Electronica } Universitat de Barcelona } Avgda. Diagonal 645-647 } 08028 Barcelona } Spain } Tel: 34 3 4021139 } 34 3 6683373 } Fax: 34 3 4021148 } E-mail: arbiol-at-iris1.fae.ub.es
-- Robin E. Schaeublin Fusion Technology - Materials Group CRPP - EPFL, 5232 Villigen - PSI, SWITZERLAND Tel : + 41 56 310 40 82 Fax : + 41 56 310 45 29
Devitrification can be identified using SEM (with appropriate precautions to eliminate charging). Sometimes, the devitrified areas grow radially from a point of nucleation. In any case, there is crystallization of the glass that will appear different from organic growth. Devitrification is a very slow process at room temperature which would be a reason to expect to find it only on very old optics.
One of our users here at UC Berkeley is looking for a cryo-TEM that would be available for her to use. There don't seem to be any that we know of here on campus. I think she'd be willing to travel to use it. Please let me know if there are any kind hearted souls out there with one that she could use. She's a graduate student in Chemistry studying surfactants and needs to study the surfactant & it's relation to its' solvent.
Thanks!
Sad because we don't have cryo,
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Please read the excellent references suggested. The Tokayasu method is wh= at we recommend to our students. Please be aware that there are so many smal= l but deadly details that will kill you. Here are : 1. If you try to rush your sucrose prep by heating you will probably not end up with good sucrose.
2. If you take more than 30 seconds to mount the tissue on the cryopin before plunging in LN2 you will probably not have good sectioning characteristics also due to molarity change.
You can get very frustrated very quickly with wrong conditions. With correct conditions even novices get beautiful 70nm sections.
A last hint, if you mince your tissue in buffer and make them a pyramidal=
shape this eliminates the need for trimming in the cryoultramicrotome. Ev= en though we have two sets of trimming tools it is usually not necessary to use them.
Good luck, contact our applications staff off-line for detailed help.
some undergrads and i are trying to prep some rabbit leg bone and growth plate for sem observation. any suggestions on how we can fix, dry, and otherwise prep the pieces?
thx! brian
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
I thought the idea was to have some fun. Come on ladies and lighten up!! No offense was meant and none should be taken. I hate it when everyone starts bickering about some trivial point which has really nothing to do with the original subject. I subscribe to this list serve in order to hear the usual intelligent exchange of advice and ideas on topics related to microscopy. The gentleman has proposed an entertaining contest for our entertainment and amusement. He playfully described an image which he thought would be an illustrative example of the type of imaging he was refering to. Nothing lewd or offensive was implied in so far as I can tell. The image of a woman in a bathing suit (with or without larvae) has never been offensive to me. Come on sisters, lighten up and have some fun already!
Now, can we get back to microscopy topics and not take off on another tangent!
Anne Marie Cooper
On Fri, 27 Feb 1998, Elinor Solit wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Did I miss something? What is fun about an image of a partially nude } woman swimming in a sea of larvae? } } There is much that could be said about this, but suffice my own comments } to be: rethink your offer, Mr. Editor, and try rephrasing in a more } respectful manner. } } Elinor Solit } The Microscope Book } }
Most TEMs these days use a side-entry goniometer sample holder. For a simple reality check on the (sample rod) tilt axis, gently press on the end of the sample holder in the microscope and observe the image deflection on the viewing screen. By the way, the common standard sample for measuring image/diffraction pattern rotations is MoO3 (available from most microscopy supply mail-order outfits).
Larry Thomas Washington State University thomas-at-mme.wsu.edu _______________________________________________________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Precise information about the goniometer tilt direction is important for many TEM applications. Historically alpha-MnO2 crystals have been used to measure the rotation angle between images and diffraction patterns in TEMs. However, for stereo analysis one needs to know the absolute tilt axis direction relative to the micrographs. I have used Kikuchi line motion to get this information. I am wondering what methods are used by others.
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I am a molecular biologist and a novice microscope user. We have an inverted phase contrast visible and fluorescence microscope used for tissue culture work. My co-workers have taught me how to focus and see cells but I don't have an understanding of what I am doing. What beginner books are available that teach basic microscope techniques for both visible and fluorescence microscopy? Are there video tapes that teach these microscopic techniques? I am starting to do tissue culture research and will be using fluorescence to study transfections and cell membrane labeling. Any advice an learning microscope use and techniques will be appreciated. Thanks Dennis P. Smith smith_dennis_p-at-lilly.com
We talked in the listserver about the way to get ultra-thin cryosections onto a grid with the method of Tokuyasu by use of a highly concentrated sucrose. We want to use cryosections of plant material for the elemental analysis, and can not use an osmoticum such as sucrose or another liquid. Is there anybody with a tricky way to get these sections?
Heike Dr. Heike Buecking University of Bremen, UFT Plant Physiology and Plant Anatomy Leobener Str. D 28359 Bremen Germany TEL: +49-421-218-2954 or TEL: +49-421-218-7283 FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de
} } I am a molecular biologist and a novice microscope user. We have an inverted } phase contrast visible and fluorescence microscope used for tissue culture } work. My co-workers have taught me how to focus and see cells but I don't have } an understanding of what I am doing. What beginner books are available that } teach basic microscope techniques for both visible and fluorescence } microscopy? Are there video tapes that teach these microscopic techniques? } I am starting to do tissue culture research and will be using fluorescence to } study transfections and cell membrane labeling. Any advice an learning } microscope use and techniques will be appreciated. } Thanks Dennis P. Smith smith_dennis_p-at-lilly.com
Two books that I like and use to teach undergraduates are:
A.J. Lacey (ed) (1989). Light Microscopy in Biology: a Practical Approach. IRL Press, Oxford ISBN 0-19- 963037-2
Bradbury, S. (1989) An Introduction to the Optical Microscope. Oxford U Press/Royal Microscopy Society. ISBN 0- 19- 856419- 8.
The Royal Microscopy Society has a number of other books in the series on specific topics such as fluorescence, immunocytochemistry, TEM, and others. While there may be more current or specific instructions for your particular application, these books provide excellent background material for the principles involved, and are oriented toward biologists.
Gary Radice Department of Biology University of Richmond
I have an EDS spectrometer and analyzer for anyone who is willing to pay for shipping costs. The EDS is an EDAX with a BE window that was fitted to a Philips 420. The analyzer is a PGT System IV complete with manuals and high end Canbera amplifier. Everything still works fine--a real work-horse. The detector efficiency has been well calibrated, so it's ready to do full quantitative analysis for some elements.
I am willing to let go of them separately, but they are best as a matched set. E-mail if you are interested.
Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets Johns Hopkins University Baltimore, Maryland 21218 USA Phone: (410) 516-8342 Fax: (410) 516-7933 e-mail: klivi-at-jhu.edu
Did you miss part of the workshop? Up the mountain maybe?!
When we were doing this in our lab, we made a simple support for the grids. It consisted of small brass plates (50 x 20 mm). They were originally a door hinge, sawn apart along the joint. The bottom part had three brass strips epoxy-glued underneath to locate it on the knife holder. On top of the plate was a rectangular wire frame, made from a paper clip, epxied down, this was to prevent grids from moving too far from the central area.
Sections were manouvered onto carbon/formvar coated grids with an mounted eyelash (that is the fun bit!). Then pressed down with the Teflon(?) part of the Reichert cryo-tools kit.
Then a lid was applied. This is the other brass plate, with a wire frame that fits around that on the base plate, for location purposes. The top plate has a simple wire handle, attached to two screws which come up through the original screw holes in the hinge. Each projecting screw has two nuts. The wire of the handle is wrapped around the screw and clamped between the nuts. This lid was screwed down with two further screws for transport, in order to keep the grids and the assembly together.
The device was then moved into a metal container with a lid in the cryobox of the microtome, then into a styrofoam box with LN2. The screws attaching the lid were then removed. The whole assembly was then transferred to an elderly vacuum station (a modified Polaron freeze fracture unit, where the knife handle was adapted to lift the cover off the box when a good vacuum was attained. Freeze drying was by simple sublimation with P2O5 powder in the chamber and a LN2 trap above the diff. pump.
Maybe this will give you some ideas. Expensive accessories!
Best wishes - and fond memories of a good workshop !!
We have a 160 liter Dewar that has not been used in years (at least 8). Our plan is to put 30 or 40 liters in it and then weigh it every day to quantify boil off rate. Can anyone tell me the density of LN? Is there a better way to determine the efficacy of a LN storage device? What are acceptable boil off rates for large Dewars (0.1-0.5 liter/day)? Can Dewars with a soft vacuum be refurbished? Additionally, compressed gas cylinders have to be tested for safety every 5 (?) years. Do LN Dewars?
TIA
Bob
Dr. Robert R. Wise, Director UW-Oshkosh EM Facility Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
Have you contacted your local compressed gas distributors? The local suppliers we use (Airco, Matheson, Middlesex Welders Supply, et al) are very knowledgeable.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
Thank you to all those who have replied and will reply to my request for a cryo-TEM.
Yes, we've checked with the folks at Donner & that TEM is booked most of the time plus I think there's a lengthy training period. But we will check again just in case there's an opening. Thanks for the reminder on that scope.
Now for the grammar police, sheesh, gimme a break! Can't a person make a mistake once in a while? Try writing a note with a lot of people coming up to you at once & see what happens.
Anyway that's my rant. But I do feel that this is not the forum for correcting grammatical errors. If you have something to contribute, please do, but lay off the nit-picky little things. It seems like this group is getting too serious. It must be grant writing time or something, or is it El Nino.....
Crabby this morning,
Paula
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I have an EDS spectrometer, but no electronics, a PGT device with thin window that was mounted on our Philips 410, for sale.
If interested, contact me.
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
We have one Philips 300 TEM in working order available for anyone who will pay for its removal from the premises. Anyone who is interested please call Dr. W.E. Mushynski at McGill University - phone 514-398-7286, e-mail address mushynski-at-medcor.mcgill.ca. Please do no reply to the listserver.
Thanks,
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca Pat Hales McGill University Department of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
Does anyone know of a plugin available for AdobeShop that will allow you to perform line profiles and print them out?
We are interested in studying cross-section samples of epitaxial grown InGaAsP layers using a line profile routine.
thanks in advance
Fred email:eoptics-at-mcmaster.ca
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
Back in mid-January there was a discussion about raster versus defocussed beam analysis. The question was raised whether the beam current was uniform across a defocussed beam, or more like a normal distribution.
The object of either method is to get a true average composition of the area illuminated by the beam. I have tested this by dragging a very small inclusion across the area of a defocussed beam. If the current density is the same everywhere, one would assume a flat-topped curve of counts versus distance, with the steepness of flanks depending only on the size of the "probe," which in this case is the size of the inclusion. I used a small gold inclusion in pyrite as test sample.
I didn't have ideal conditions for this, but within the limitations of what I could do I have verified that the current density across a defocussed beam is indeed approximately constant, although it remains to be seen whether this depends on particular conditions (diameter, current, etc.). Anyone who would like to know the details is welcome to e-mail me directly.
Alfred Kracher ----------------------------------------- Geological Sciences 253 Science I Iowa State University Ames, IA 50011-3212 mailto://akracher-at-iastate.edu http://www.public.iastate.edu/~akracher vox:515 294 5439 fax:515 294 6049 -----------------------------------------
I am having a recurring problem with charging during SEM imaging of coal ash. The ash specimens (chunks of porous, sintered material from boiler walls) are embedded in epoxy and then sectioned and polished for examination. The epoxy used is a lower-viscosity, two-component resin sold by a major metalography supply company for impregnation of porous materials. A gold coating has been routinely used.
I would appreciate any help and suggestions with this problem.
Everett Ramer Federal Energy Technology Center U.S. Department of Energy Pittsburgh, PA ramer-at-fetc.doe.gov
Embedded specimens may well have a gold coat across the surface but is th= e complete mount earthed?
Run a silver Dag earth from the top surface down to the base, or stub, as= a gold coat is usually not enough to conduct down to the base of the stub; the plasma does not usually run too far down the side in my experience..
Lower the kV down to about 5 to be sure of not running at too high a voltage.
Why do you mount and section, as I would handle this specimen my, casting= a very small amount onto a double sided carbon tape and running at 5kV?
If we can help further please do not hesitate to ask.
Steve Chapman
Senior Consultant E.M. Protrain., Oxford, UK Tel & Fax 44 (0)1844 353161 web site at http://ourworld.compuserve.com/homepages/protrain
I am trying to determine or calibrate magnification on my scope using a grating replica which has the following formula:
mag = distance in cm between limiting lines (X) 21,600 lines/cm number of spaces between limiting lines
If you have experience with such, what is meant by the denominator "number of spaces between limiting lines"? As I view the replica I only see parallel lines. I have a call in to the manufacture but am getting impatient. Thanks.
Bill Meek Meekwd-at-osucom-fs02.ocom.okstate.edu Department of Anatomy and Cell Biology OSU-College of Osteopathic Medicine 1111 W. 17th St. Tulsa, OK 74107 918-561-8258 918-561-8414 Fax
} I am having a recurring problem with charging during SEM imaging of coal } ash. The ash specimens (chunks of porous, sintered material from boiler } walls) are embedded in epoxy and then sectioned and polished for } examination. The epoxy used is a lower-viscosity, two-component resin } sold by a major metalography supply company for impregnation of } porous materials. A gold coating has been routinely used.
This should not happen if coated with gold.
Check that the coating completely covers the specimen and is in contact with the stub which is then in contact with the stage. Also check that the stage is properly grounded. We often paint a line of conductive paint from the stub onto the polished face of the specimen to ensure continuity to ground.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Your accuracy is improved by taking multiple lines, i.e. a longer distance on the micrograph. For example, if eleven lines are present in your micrograph, then there are ten spaces. Just make sure that you start and end on the same relative side of the width of the lines. What you are really counting is the number of repeat distances in the grating that you are using. -Scott Walck
I am trying to determine or calibrate magnification on my scope using a grating replica which has the following formula:
mag = distance in cm between limiting lines (X) 21,600 lines/cm number of spaces between limiting lines
If you have experience with such, what is meant by the denominator "number of spaces between limiting lines"? As I view the replica I only see parallel lines. I have a call in to the manufacture but am getting impatient. Thanks.
Bill Meek Meekwd-at-osucom-fs02.ocom.okstate.edu Department of Anatomy and Cell Biology OSU-College of Osteopathic Medicine 1111 W. 17th St. Tulsa, OK 74107 918-561-8258 918-561-8414 Fax
Liquid nitrogen density at -195.8C is 0.808. Its worth noting that a litre of liquid turns into 800 litres of gas. The fairly robust smaller (25 or 30 l) dewars use about a litre per day. Similar size storage flasks as used in the semen industry would last about six months - full to empty. Those flasks have fewer (weaker) contact points at the neck and the low parts of the flask and should not be used for pouring LN2.
I guesstimate that a 160 l dewar would use at least 3 litres per day. Good vacuum is essential to minimise LN2 losses. All dewars I have seen can be repumped. I had an adaptor made up that sealed against the chamber intake of an evaporator (used without bell jar) a vacuum hose led to the adaptor valve which fitted over the vacuum "bung" of the dewar. With different end adaptors that device was also used for pumping EDS dewars.
Final vacuum attained is improved if some boiling water is poured into the dewar a few hours before terminating pumping. The next paragraph describes the pumping device. If you want to proceed with this exercise I would be happy to email a crude drawing to explain the function of the adaptor, sorry no engineering drawings; its too long ago.
You will find that there is a screw plug somewhere on your dewar which can be removed; its function is to protect the vacuum plug below. The vacuum plug has a threaded centre hole and the "adaptor valve" was designed to include a shaft which had a matching thread on the end and protruded through the end of the valve ("O" ring sealed). A short crossbar permitted engaging the thread and pulling the bung out by about 25mm. Tape was used to prevent the shaft from being sucked back in. The front of the valve (with the threaded shaft) had a sealing nut which would mount over the top of dewars vacuum connection.
Quite a useful device for a big lab with a number of dewars. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} We have a 160 liter Dewar that has not been used in years (at least } 8). Our plan is to put 30 or 40 liters in it and then weigh it every day } to quantify boil off rate. Can anyone tell me the density of LN? Is there } a better way to determine the efficacy of a LN storage device? What are } acceptable boil off rates for large Dewars (0.1-0.5 liter/day)? Can Dewars } with a soft vacuum be refurbished? Additionally, compressed gas cylinders } have to be tested for safety every 5 (?) years. Do LN Dewars? } } TIA } } Bob } } } Dr. Robert R. Wise, Director } UW-Oshkosh EM Facility } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } www.uwosh.edu/departments/biology/wise/wise.html } } }
} I am trying to determine or calibrate magnification on my scope using a } grating replica which has the following formula: } } mag = distance in cm between limiting lines (X) 21,600 lines/cm } number of spaces between limiting lines } } If you have experience with such, what is meant by the denominator } "number of spaces between limiting lines"? As I view the replica I only } see parallel lines.
You literally count the spaces between the lines. For example,
| | | | | shows 4 spaces
| | | | | | | | shows 7 spaces
I suppose you could count the lines and subtract one but it is presumably easier to count (and possibly tick off or mark) the spaces rather than the lines.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
in the past there have been several threads dealing with charging problem= s in SEM and their remedies.
I was wondering if it were possible to take a "common" embedding resin an= d mix in a certain amount of graphite powder, thereby generating a pseudo-conductivity in the resin itself. =
"Pseudo-conductivity" because there will probably be microscopic, resin filled spaces between the graphite particles that would act as an isolato= r to lower voltages (e.g. Ohmmeter), but when exposed to a high voltage potential, the situation may change...
Has anyone experimented with such a home-brew "conductive- resin"? If so,=
what were your experiences? How does, for example, the graphite powder affect the curing of the resin? How much powder is needed to make a commo= n two-component resin conductive (relation of volume)? Is precipitation a problem?
If, by chance, this is something of common knowledge, then please forgive=
At 04:14 PM 3/3/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Several manufacturers make conductive resins to help with this problem, as I found out from this list when I had a related problem last year. I will try to locate my saved files and forward them to you, but querying the various EM suppliers may give you some good leads. A couple users suggested mixing fine carbon powder with the resin.
I have no direct experience with the effectiveness of these resins, yet, since I was primarily gathering information for a third party.
The other things you might try are SEM operations things that you probably already know about. I'll throw some in anyway, since I don't know what your experience level is. Please forgive me if I'm being obvious. Charging can be minimized by using lower accelerating voltages and smaller condenser spot sizes. Some instruments are designed to operate effectively at 1 kV or lower, especially if they can integrate several scans to form the final image.
That's another way, incidentally. If you have a scope that can digitally integrate consecutive scans to make a clean image of many noisy ones, this can drastically reduce charging effects, by reducing the amount of time the beam remains on a particular part of the sample during each scan.
If you have a "plug" of ash surrounded by resin, using conductive paint "bridges" from the plug to the mounting stub can help. Might help anyway, even if the ash is dispersed throughout the resin. Make sure that the specimen is mounted to the stub with enough conductive adhesive to form a good path.
And then, use the "environmental" or "variable pressure" capabilities of the scope, if it has them. I usually start at the lowest pressure available, then rack it up one notch at a time until charging is manageable. Usually 1-7 Pa of pressure will do it.
Hope something here helps.
Randy Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
A colleague needs to know the mean free path of 100keV electrons in ZrO2. He is trying to determine the absolute thickness of samples examined by PEELS. The EL/P program calculated a relative thickness in terms of the number of mean free paths but we want to convert this to thickness in nanometres. Any help would be appreciated. cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
I would suspect that in the case of a bulk insulator with a thin conducti= ve coating, that some charge would be injected into the bulk material (a few=
microns deep, depending on atomic number and beam energy.) Even though surface charges may drain away, the bulk charge would remain.
Dear list I have just carried out some EDX analysis of some old (?18th Century) ceramic? floor tiles. Does anyone know how I might use the compositional analysis to perform some form of dating? The tiles come from the cellars of a mansion in Cheshire UK. As a biologist I know nothing about these things! Many thanks
Chris Gilpin Biological Sciences EM Unit G452 Stopford Building Manchester University Oxford Road Manchester M13 9PT phone +44 161 275 5170 Fax +44 161 275 5171
Have you tried using a conductive epoxy, you can make your own by mixing some carbon powder in to your normal resin. This, together with gold coating should reduce the charging.
Another idea could be to ensure that contact between the resin and sample holder is maintained. The carbon in the epoxy should help this too.
Jo Hunt Kodak Ltd Harrow Middlesex HA1 4TY ENGLAND (jshunt-at-kodak.com)
Hi all I am a novice of this list, so please write me is there is some FAQ to avoid starting again old threads. By the way, is there a consolidated way of protecting a SEM chamber from boost pressure coming from N2 bottle during venting? (this may occur if someone accidentally moves the low pressure stage on the bottle)
Thank you in advance Dr. Eng. Edoardo Bemporad, Ph. D. Assistant Professor of Materials Science University of Rome "Roma Tre" (Italy) Dipartimento di Ingegneria Meccanica e Industriale (Department of Mechanical and Industrial Engineering) Via della Vasca Navale 79 - 00146 Rome, Italy Tel: +39 6 5517.3293 Fax: +39 6 5517.3256 LIME Lab (InterDipartimental Laboratory of Electron Microscopy) Tel: +39 6 5517.3200 E-Mail:bemporad-at-uniroma3.it
} I am trying to determine or calibrate magnification on my scope using a } grating replica which has the following formula: } } mag = distance in cm between limiting lines (X) 21,600 lines/cm } number of spaces between limiting lines } } If you have experience with such, what is meant by the denominator } "number of spaces between limiting lines"? As I view the replica I only } see parallel lines.
It seems to me that here we have here a rather pedantic way of expressing the fact that, if one is going to measure say 10 spaces, one has to count the lines from #1 at one end to #11 (i.e. n+1) at the other.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Within the frame of a European research project on coatings for Ultra violet applications, several post-doc positions are available, two of them related to TEM techniques. The host center for this two ones is University of Barcelona and the main objectives of the work is the structural and compositional characterization of UV-coatings.
Is someone is interested, please contact Dr. F. Peir=F3 at
paqui-at-iris1.fae.ub.es
Please find more information visiting the web site:
www.lzh.de/tmr/default.htm
*******************************+ Francesca Peiro
EME, Enginyeria i Materials Electronics Dpt. Fisica Aplicada i Electronica Universitat de Barcelona Avda. Diagonal 645-647 08028 Barcelona, Spain
Whilst we are on the subject of conductive resins, I am looking at embedding some ceramic powder particles in resin prior to ion beam thinning (since they are far too large to be electron transparent without thinning). I was thinking that embedding in a conductive resin may be a good idea. Some have suggested mixing carbon (carbon black I assume) with the resin. I have heard in the past about use of a silver loaded resin. Is this readily available? Is it horribly expensive? Does anyone know of a vendor who sells this in the UK?
Also, some epoxy resins have been optimised for materials work so that they can be cured to a high hardness value. How do silver or carbon loaded epoxies compare with this?
Thanks for any help you can give.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
I am not sure what sort of 160l dewar system you have but the two that we use have a pressurised delivery system using a cluster of valves at the top. The important thing to bear in mind is that it is pressurized and so care should be taken. Our system has a spring loaded pressure relief valve and a burst disk relief valve. If your system is like this then I would STRONGLY RECOMMEND that you have the valves checked before putting liquid nitrogen in, especially as it hasn't been used for eight years.
You may also need to consider how much other liquid nitrogen is lost than by evaporation in the 160l dewar. You will lose some in pumping it out and probably lose a small amount when it no longer pumps out (ie level is too low). The best we have ever had is just under 5 x 25l dewars filled from a fresh 160l dewar.
Good luck
Malcolm Haswell Electron Microscope Unit University of Sunderland UK ----------
Dear Chris,
If your interest in these tiles is to record their origin and history, you may get better results from macrostructural evidence. Details such as size, color, glaze, inclusions, etc. can tell a very rich story to someone who can read those clues. You may have an expert on campus right now. I'd take a few of the tiles to the art or history department. I wouldn't be surprised if they could pin-point the origin to a specific kiln!
Harold J. Crossman Senior Scientist OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
Chris Gilpin wrote: } I have just carried out some EDX analysis of some old (?18th Century) } ceramic? floor tiles. Does anyone know how I might use the compositional } analysis to perform some form of dating? } The tiles come from the cellars of a mansion in Cheshire UK. } As a biologist I know nothing about these things! } Many thanks
Dear Chris, The best way to date ceramic materials is probably to use thermoluminescence dating. For further information you may contact the Nordic Laboratory for Thermoluminescence Dating which is situated here at Risoe. The phone no. is +45 46 77 59 75.
Best wishes, Joergen
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
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************* Heide Schatten, Ph.D. Associate Professor Department of Veterinary Pathobiology Veterinary Medicine Building University of Missouri-Columbia 1600 E. Rollins Street Columbia, MO 65211 TEL: (573) 882-2396 FAX: (573) 884-5414 alternative e-mail: hschatte-at-facstaff.wisc.edu
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Dewars can be recharged with insulation gas and minor leaks fixed in many cases. It really pays to have the dewar checked out IF it does not hold nitrogen. When we had an insulating gas leak in the walls of ours, the dewar frosted up noticably and there was no doubt we had a problem. We took it to an appropriate regional vendor and had it repaired for a fraction of the cost of replacing.
Debby Sherman, manager Microscopy Center in Agriculture Purdue University West Lafayette, IN 47907
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I used to have a similar problem with 1 inch diameter, polished epoxy mounts of printed circuit boards. I solved the problem by sputtering with Au-Pd and grounding the mount by painting a line from the polished surface of the mount to the specimen holder using carbon paint ("aquadag" sticks in my memory). Alternatively, gold sputtering with a silver paint ground might work. I really think the key was painting a good connection to the specimen holder.
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At 04:14 PM 3/3/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Several manufacturers make conductive resins to help with this problem, as I found out from this list when I had a related problem last year. I will try to locate my saved files and forward them to you, but querying the various EM suppliers may give you some good leads. A couple users suggested mixing fine carbon powder with the resin.
I have no direct experience with the effectiveness of these resins, yet, since I was primarily gathering information for a third party.
The other things you might try are SEM operations things that you probably already know about. I'll throw some in anyway, since I don't know what your experience level is. Please forgive me if I'm being obvious. Charging can be minimized by using lower accelerating voltages and smaller condenser spot sizes. Some instruments are designed to operate effectively at 1 kV or lower, especially if they can integrate several scans to form the final image.
That's another way, incidentally. If you have a scope that can digitally integrate consecutive scans to make a clean image of many noisy ones, this can drastically reduce charging effects, by reducing the amount of time the beam remains on a particular part of the sample during each scan.
If you have a "plug" of ash surrounded by resin, using conductive paint "bridges" from the plug to the mounting stub can help. Might help anyway, even if the ash is dispersed throughout the resin. Make sure that the specimen is mounted to the stub with enough conductive adhesive to form a good path.
And then, use the "environmental" or "variable pressure" capabilities of the scope, if it has them. I usually start at the lowest pressure available, then rack it up one notch at a time until charging is manageable. Usually 1-7 Pa of pressure will do it.
Hope something here helps.
Randy Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
Fred-- I don't know of a plug-in, but I solved the problem another way. If you export the image (256 grey levels) as encapsulated post script (*.eps), the raster data are hexadecimal pairs. You can read this, with formatted input, in Fortran with a Z mask converting the data to integers. You can then do line length calculations, perform any sort of spatial averaging you want, calculate fractal dimensions, etc. If you are keen to analyze the 2-D image, you can use this as grid data input for a Geographic Information System (GIS) program. I use ArcView with the Spatial Analyst extension for contouring, custom color coding...If you use these data as feature input to ArcView, you can do trend surface analysis, and on and on. Contact me if you want to know more about the Fortran Routines... jptmvl-at-mailbox.syr.edu (Dave Johnson, Dept. Chem, SUNY College of Environmental Science and Forestry).
With regard to thermoluminescence (TL) dating of fired ceramics, here are links to a number of pages on the topic recently provided me by Daniel Richter of McMaster University in Ontario.
The method is used commonly to date fired ceramics and hollowcast metal sculpture containing remains of ceramic vestment. A sample is heated to release (luminesce) free electrons trapped in lattice defects in clay and other minerals since the object was last fired or heated to high temperature. Luminescence is measured to determine an approximate age.
http://www.aber.ac.uk/~ggd/ Luminescence Dating Laboratory Aberystwyth
http://users.ox.ac.uk/~uzdh0045/ Luminescence Dating at the Research Laboratory for Archaeology and the History of Art, Oxford University
http://weber.u.washington.edu/~anthro/departmentinfo/TLLabInfo.html The University of Washington Luminescence Dating
http://is.dal.ca/~digs/t-intro.htm Dalhousie TOSL Laboratory: TL, OSL, ESR dosimetry & dating
http://geosun.geog.susx.ac.uk/Luminescence/ldqg.html Luminescence Dating and Quaternary Geomorphology
Good luck with the project.
James Martin Analytical Services and Research Williamstown Art Conservation Center 225 South Street Williamstown, MA 01267 tel: 413.458.5741 fax: 413.458.2314
Don't know the answers to most of your questions, but the back of a business card from a gas supplier rep says that 1 gal = 93.11 SCF (standard cubic feet) = 6.745 lbs
Perhaps your LN2 supplier can be of assistance?
Jim Passmore Analytical Chemist Cryovac North America james.passmore-at-grace.com
---------- } From: wise } To: Microscopy } Subject: LN2 questions } Date: Tuesday, March 03, 1998 6:59AM } } { {File Attachment: LN2QUEST.TXT} } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Chris, I suggest you contact the Research Lab for Archaeology at Oxford - someone there should know. Don Marshall
} Dear list } I have just carried out some EDX analysis of some old (?18th Century) } ceramic? floor tiles. Does anyone know how I might use the compositional } analysis to perform some form of dating? } The tiles come from the cellars of a mansion in Cheshire UK. } As a biologist I know nothing about these things! } Many thanks } } } Chris Gilpin } Biological Sciences EM Unit } G452 Stopford Building } Manchester University } Oxford Road } Manchester } M13 9PT } phone +44 161 275 5170 } Fax +44 161 275 5171
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
I used a diver's demand valve to stop the N2 flow. The demand valve closes when the instrument stops sucking - just like a diver only gets air while sucking. As for removing the pressure regulator. I would expect that taping the control and a sign "do not touch" would suffice. Lesser offenders have suffered capital punishment. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} Hi all I am a novice of this list, so please write me is there is some FAQ to avoid starting again old threads. } By the way, is there a consolidated way of protecting a SEM chamber from boost pressure coming from N2 bottle during venting? } (this may occur if someone accidentally moves the low pressure stage on the bottle) } } Thank you in advance } Dr. Eng. Edoardo Bemporad, Ph. D. } Assistant Professor of Materials Science } University of Rome "Roma Tre" (Italy) } Dipartimento di Ingegneria Meccanica e Industriale } (Department of Mechanical and Industrial Engineering) } Via della Vasca Navale 79 - 00146 Rome, Italy } Tel: +39 6 5517.3293 } Fax: +39 6 5517.3256 } LIME Lab (InterDipartimental Laboratory of Electron Microscopy) Tel: +39 6 5517.3200 } E-Mail:bemporad-at-uniroma3.it } } } } }
EDX (EDA, EDS and WDS) analysis gives elemental composition only. The only way you could conclude anything about age, would be based on specific knowledge about glazes which become available only after a certain date. In paintings the now very common TiO2 white pigment only arrived this century. If you find Ti in a Ruben's its a fake. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} Dear list } I have just carried out some EDX analysis of some old (?18th Century) } ceramic? floor tiles. Does anyone know how I might use the compositional } analysis to perform some form of dating? } The tiles come from the cellars of a mansion in Cheshire UK. } As a biologist I know nothing about these things! } Many thanks } } } Chris Gilpin } Biological Sciences EM Unit } G452 Stopford Building } Manchester University } Oxford Road } Manchester } M13 9PT } phone +44 161 275 5170 } Fax +44 161 275 5171 } } }
} A colleague needs to know the mean free path of 100keV electrons in ZrO2. } He is trying to determine the absolute thickness of samples examined by } PEELS. The EL/P program calculated a relative thickness in terms of the } number of mean free paths but we want to convert this to thickness in } nanometres. Any help would be appreciated. cheers,
For a handy rule of thumb, consider assuming that the inelastic mfp in most things is about 25 ug/cm^2 for 100kV electrons (that's in "micrograms per square centimeter"), and about 50 ug/cm^2 for 300kV electrons. Dividing by the density of your ZrO2 would then give you a 1st-order estimate of its thickness. The EELS experts in this forum can probably provide a more precise value for this in your case, as well as information on its (I think relatively weak) dependence on atomic number.
This strategy also provides a rule of thumb for experimentally determining the mass of material in your field of view, since no assumptions about density are needed to multiply by the projected area (in cm^2) of your specimen!
I aggree with all the other contributions, but "chunks of porous, sintered material . . . " makes me think that the major problem is the highly perforated nature of the specimen. Sputter coaters scatter the gold much more than evaporators, but most of the material is applied to the top face and does not enter from the near horizontal the inside of those numerous, on the polished surface opened holes. I expect that if Everett was to inspect the specimen on a steep angle under a high power disecting scope, he would find the Au film rather discontinious. A partial remedy, which combined with other methods proffered might give success would be to coat these samples two or three times. Angle the specimens at about 60 degees from the horizontal and apply that tilt for each coating from an opposing direction.
Another point: If you have a good (I liked the Robinson best) backscattered e detector, it may give sufficient resolution and is much less prone to charging effects. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} I am having a recurring problem with charging during SEM imaging of coal } ash. The ash specimens (chunks of porous, sintered material from boiler } walls) are embedded in epoxy and then sectioned and polished for } examination. The epoxy used is a lower-viscosity, two-component resin } sold by a major metalography supply company for impregnation of } porous materials. A gold coating has been routinely used. } } I would appreciate any help and suggestions with this problem. } } Everett Ramer } Federal Energy Technology Center } U.S. Department of Energy } Pittsburgh, PA } ramer-at-fetc.doe.gov }
Job Title/Number Research Specialist/11410 Opening Date: 3/5/98 Closing Date: 3/11/98 Categorization: Regular Classified Staff Benefits Eligible: Yes =09 Hiring Range: $22,602-$30,459 FTE: 0.50=09 Reports To: Dave Bentley Department: Arizona Research Labs, Biotechnology Imaging Facility
Position Summary: Person hired will engage in service work and other support activities in the Biotechnology Imaging Facility. Position requires manual labor including lifting 50 lbs to 4 feet high, carrying 50 lbs 100 feet, moving compressed gas tanks weighing 200 lbs, lifting 20 lbs to 6 feet, and opening liquid nitrogen tank valves (20 ft lbs of torque).
Duties and responsibilities include:
=B7 Perform transmission electron microscope service work of moderate difficulty for facility clients under general supervision. =B7 Perform scanning electron microscope service work for clients. =B7 Assist with laboratory maintenance. =B7 Teach users in proper operation of Facility equipment. As required, assist users with operation of the equipment during learning phase. Assist with other designated instructional tasks. =B7 Acquire, manipulate, measure, label and print computer images. Maintain imaging computer and ancillary equipment. Back up data files, install and update programs. Instruct users on proper operation of computer and of image analysis programs. =B7 Assist users with operation of the Facility equipment, or direct questions to another staff member, as appropriate. Troubleshoot problems. Assist in maintenance and repair of Facility equipment. =B7 Supervise lower level staff as necessary. =B7 Perform administrative duties such as data entry, billing, preparation o= f letters and handouts, sorting, collating, photocopying, phone answering, ordering, etc.=20
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These big dewars are not usually made with a vacuum but are insulated with a powder. In this case there should be no problem after sitting around for 8 years.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
At 11:17 PM 3/3/98 -0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm not aware of any direct method of using compositional analysis for dating, but having done graduate work in archaeology I can tell you that archaeologists are the folks you need to consult. Historical archaeologists usually have access to data bases of artifact types that can be used to trace date of manufacture, etc. It's possible that some people have compiled data concerning compositional analysis of various types of items, but I don't know. My guess is that identification and dating will end up being done by other characteristics of the tiles, i.e., appearance, makers marks, color, and so on.
Best wishes, Randy Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
I would also be interested in any thecnique that people have that incorporates carbon or graphite or any conductive substance into Spurr's or any of the other epoxy resins. We have plant biologists who are making replicas of their plants using dental impression molds & then making a replica of their plant using Spurr's, I thought this might be interesting to try.
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
As noted, some (bulk) conductive resins are commercially available. Most seem to be thermal set (hot press). Conductive cold set resins sem to be more rare. I have experimented with adding conductive material to multi-part cold setting resins. Suggest lampblack as posibility if you use carbon - fine particles. I did not like some of the characteristics of carbon and have used zinc dust (very fine). A very "rich" loading is required. DO BEWARE of any potential reactions between the resin and whatever metal dust used.
None are satisfactory if you must examine an edge (mount specimen interface).
In all cases charging resin islands abound.
Sometimes I carefully (carbon) paint around the specimen using a low mag stereo scope & 0000 sable brush. Best done before too much coffee {g} .
Have also used low melt point metal (ie Wood's metal, Cereloy) in a hot press mount system. Down - side is soft mount.
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Dear All,
in the past there have been several threads dealing with charging problems in SEM and their remedies.
I was wondering if it were possible to take a "common" embedding resin and mix in a certain amount of graphite powder, thereby generating a pseudo-conductivity in the resin itself. "Pseudo-conductivity" because there will probably be microscopic, resin filled spaces between the graphite particles that would act as an isolator to lower voltages (e.g. Ohmmeter), but when exposed to a high voltage potential, the situation may change...
Has anyone experimented with such a home-brew "conductive- resin"? If so, what were your experiences? How does, for example, the graphite powder affect the curing of the resin? How much powder is needed to make a common two-component resin conductive (relation of volume)? Is precipitation a problem?
If, by chance, this is something of common knowledge, then please forgive my ignorance regarding the subject.
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Hey All,
Is there anyone who will be attending the ICEM in Mexico that SCUBA dives? Are you planning on taking a day or so to dive? If so, could you please email me off line? Perhaps we could get together and go diving. I would much rather dive with someone from the group, who I can talk to ahead of time, than be thrown together with a total stranger on a dive boat.
Thanks,
David Bell Millipore Corporation 80 Ashby Road Bedford, MA 01730 1 800 221-1975x2108 David_Bell-at-Millipore.com
The Ag material is available from a number of microscopy suppliers in the US. All I have seen, however, has rather large Ag flakes...
Along this thread... I am still waiting for the chemists to develop a thermoplastic version of intrinsically conductive plastic. Intrinsically conductive polymer is around, but all I have seen is milled thermoset - May as well use carbon or metal powders....
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Whilst we are on the subject of conductive resins, I am looking at embedding some ceramic powder particles in resin prior to ion beam thinning (since they are far too large to be electron transparent without thinning). I was thinking that embedding in a conductive resin may be a good idea. Some have suggested mixing carbon (carbon black I assume) with the resin. I have heard in the past about use of a silver loaded resin. Is this readily available? Is it horribly expensive? Does anyone know of a vendor who sells this in the UK?
Also, some epoxy resins have been optimised for materials work so that they can be cured to a high hardness value. How do silver or carbon loaded epoxies compare with this?
Thanks for any help you can give.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
4 Mar 98 13:41:05 -0600 Received: from SpoolDir by OBSNSRV1 (Mercury 1.21); 4 Mar 98 13:40:59 -0600 Received: from botstrou.bio.ou.edu by obsnsrv1.bio.ou.edu (Mercury 1.21); 4 Mar 98 13:40:59 -0600 Message-ID: {34FDAE2C.11AC-at-ou.edu}
We have a demand valve on our TEM so that venting does not over-pressure the column. Basically the valve allows N2 gas to enter the column as long as it "sees" a negative pressure. As the pressure in the column rises to atmosphere, the flow of N2 slows until it is at equilibrium with the room. The same valve is used for venting the camera and the column as the demand valve is part of the regulator on the N2 bottle.
Edoardo Bemporad wrote: } } Hi all I am a novice of this list, so please write me is there is some FAQ to avoid starting again old threads. } By the way, is there a consolidated way of protecting a SEM chamber from boost pressure coming from N2 bottle during venting? } (this may occur if someone accidentally moves the low pressure stage on the bottle)
-- ======================================================== Greg Strout Electron Microscopist, University of Oklahoma e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ========================================================
Magnification =3D total measured distance/number of squares or parallel l= ine units X ( 1/2160)0.4629um. Round the figures to the nearest 500X.
With a TEM expect an accuracy of about plus/minus 5%. With a SEM expect plus/minus 10% with a difference on a cross grating in each direction of less than 5%.
Be aware that with a TEM you must have the stage at the eucentric point and give the high voltage at least 120 minutes running before you start work, 45 minutes in a SEM; this gives the high voltage chance to settle (= HT tank heat gained =3D heat lost).
In the SEM the spot size will also have an effect upon magnification. Ha= ve you noticed how with a manual instrument you need to re focus when you change the spot size: the result a magnification change!
Need any more help please ask.
Steve Chapman
Senior Consultant E.M. Protrain., Oxford, UK Tel & Fax 44 (0)1844 353161 web site at http://ourworld.compuserve.com/homepages/protrain
} Hi all I am a novice of this list, so please write me is there is some FAQ } to avoid starting again old threads. } By the way, is there a consolidated way of protecting a SEM chamber from } boost pressure coming from N2 bottle during venting? } (this may occur if someone accidentally moves the low pressure stage on } the bottle) } } Thank you in advance } Dr. Eng. Edoardo Bemporad, Ph. D. } Assistant Professor of Materials Science } University of Rome "Roma Tre" (Italy) } Dipartimento di Ingegneria Meccanica e Industriale } (Department of Mechanical and Industrial Engineering) } Via della Vasca Navale 79 - 00146 Rome, Italy } Tel: +39 6 5517.3293 } Fax: +39 6 5517.3256 } LIME Lab (InterDipartimental Laboratory of Electron Microscopy) Tel: +39 6 } 5517.3200 } E-Mail:bemporad-at-uniroma3.it
The easiest solution is to have a 'T' piece in the line connecting the N2 cylinder to the SEM, with the leg of the 'T' open to the atmosphere. Adjust the N2 pressure so that there is a reasonable flow of N2 to atmosphere when not venting. It's not precisely controlable but it stops over pressure problems.
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
It seems that discussion of partial nudity of either sex on a bulletin would be so obviously inappropriate, maybe that is why it hasn't occurred before, except in reference to Bayard-Alpert gauges, in which case it is OK. John Mardinly Intel
The Application Deadline for the UBC 3D Microscopy of Living Cells Course has had to be delayed until March 15, 1998. (There have been some problems with my fax machine. If you received an email confirmation, everything is fine, if not please fax your application again.)
No space here to give you the whole story about this unusual international course but I can give some "coded" hints that 3D microscopists will understand:
Zeiss 560, Ted Inou=E9, Abb=E9 diffraction kits, Infinity-Optical, Bill Maso= n, Scanalytics, Paul Negulescu, Bio-Rad Micro-Radiance, SGIs, Eppendorf microinjection, Larry Keenan, Optical tweezers Cameleon, Applied Precision, Yokogawa, Jon Art, 1024/multi-photon, Sigrid Myrdal, PSI-CCD, Fluoview, Tim Murphy, GFP-transfection, Huygens, Video-rate, Time-bandwidth laser, Warren Zipfel, brain-slice, spherical aberration corrector, digital time-lapse, Ernst Stelzer, Noran-OZ, P.C. Cheng, Cell-Robotics, 3D image processing, Yu-Li Wang, ALAScience chamber, Paul Millard, Zeiss 510, Jay Margolis, Nikon PCM-2000, Clarke laser, Dan Focht, Nikon PCM-2000 (again), Intracell chamber, Astromed, Universal Imaging, Hans vanderVoort, Driftwood Confocal Contest, backscattered light, water-immersion.
They will all be in Vancouver, not just to look at, but to talk to, to work with and to use during over 40 hours of hands-on, structured labs (and an additional 20 hours for "personal" live-cell projects.)
Hope that you can join us in Vancouver this June 17-28.
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Engineering Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr.,
You might contact John Watt at Middlesex Poly--he is interested in such analytical challanges and he has some museum conservator contacts at the V and A
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Just a remark on conductive resins: they are good to embed conductive samples for surface SEM observation for example. But don't forget, they are composite materials, i.e. non-conductive resin loaded with enough conducting material (Cu, Ag whatever) to make them MACROSCOPICALLY conductive.
In microscopy application, their composite structure quickly appears and limit their usage. For example, their are useless to look at edges of conductive material because the non-conductive resin portion in-between the conductive particles will charge up exactly as pure polymer and blur out the picture.
For your ion-milling application I suspect the same holds true but I haven't try myself. So just be aware of this problem if doesn't work.
BTW they are available from any electron microscopy accessories vendor.
Jean-Marc Boichat email: jean-marc.boechat-at-chma.mhs.ciba.com EM LABS FO 5.1 phone:+4126 435 6979 fax: +4126 435 6907 Ciba research Center P.O. Box 64 CH- 1723 Marly 1 When things go wrong, don't go with them! Switzerland
Disclaimer: "nobody in this company ever cared for what I said, why would they start now".
This message echoes a previous message by PA Buffat. Indeed we have two Philips EM300 TEMs to give. One is in perfect state, and the other had a HV tank problem but works at 80 kV.
There is a possibility to get funding for the transport from the Development and Cooperation Directory of the Swiss Ministry of Foreign Affairs. If you are interested please contact me directly.
Yours,
Robin Schaublin
-- Robin E. Schaeublin Fusion Technology - Materials Group CRPP - EPFL, 5232 Villigen - PSI, SWITZERLAND Tel : + 41 56 310 40 82 Fax : + 41 56 310 45 29
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you said...
In the SEM the spot size will also have an effect upon magnification. Have you noticed how with a manual instrument you need to re focus when you change the spot size: the result a magnification change! ...................
I believe this statement is inaccurate. When spot size is changed in an SEM, you are changing the strength of the condenser lenses. This in turn effects the position of the beam cross-over and thus the portion of the beam that passes through the objective aperture and into the final (objective) lens). The final lens then must be adjusted to focus the beam crossover onto the surface of the specimen. The final lens is not an enlarging lens, like the objective lens in a TEM or light microscope. It does not have a role in magnification in an SEM. It does have a role in specimen image position, since when you change the strength of the lens, you effect the length of the spiral path of the electrons in the beam and thus can cause image rotation. Magnification in an SEM is determined by the scan coils. The scan path across the specimen is reflected on the monitor. As the monitor size is fixed, a longer scan path results in a smaller magnification, and conversely, a shorter scan path results in a higher magnification. I believe, therefore, that since no lenses are involved, magnification should be independent of focus and focus, as determined by strength of the final lens required to adjust cross-over, is effected by the change in the condenser lens strength when beam diameter (probe current) is adjusted.
Debby Sherman, manager Microscopy Center in Agriculture Purdue University West lafayette, IN 47907 sherman-at-aux.btny.purdue.edu
I agree with Jean Marc. All this excitement about conducting resins is not likely to achieve the aim, which was the elimination of charging in the SEM of DIFFICULT non-conductors embedded in resins. Consider that most elemental standards for EDS are mounted in resin blocks. Furthermore, in WDS especially much higher specimen currents are used than in normal SEM. Those standards have a 20nm, heavy carbon coating, but this is not as conductive as is the Au coating employed in SEM. Normally in analysis BS detection is used but charging in secondary mode is uncommon. If the argument was right that resin embedded non-conductors charge because of the resin, EDS and WDS would have a few additional problems. WHY THEN SHOULD CONDUCTING RESINS SOLVE CHARGING PROBLEMS IN SEM?
The difference is the type of embedded material. All standard materials used must permit a fine polish and cannot be porous or highly fractured. Such materials will have a continues conducting layer accross the surface and not charge. The original correspondent has charging problems because of the highly porous nature of his specimens. Better (angled) coatings and optimising the instrument's parameters to minimize charging will hopefully solve his problem. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** -----------------------------------------------------------------------.
Just a remark on conductive resins: they are good to embed conductive samples for surface SEM observation for example. But don't forget, they are composite materials, i.e. non-conductive resin loaded with enough conducting material (Cu, Ag whatever) to make them MACROSCOPICALLY conductive.
In microscopy application, their composite structure quickly appears and limit their usage. For example, their are useless to look at edges of conductive material because the non-conductive resin portion in-between the conductive particles will charge up exactly as pure polymer and blur out the picture.
For your ion-milling application I suspect the same holds true but I haven't try myself. So just be aware of this problem if doesn't work.
BTW they are available from any electron microscopy accessories vendor.
Jean-Marc Boichat email: jean-marc.boechat-at-chma.mhs.ciba.com EM LABS FO 5.1 phone:+4126 435 6979 fax: +4126 435 6907 Ciba research Center P.O. Box 64 CH- 1723 Marly 1 When things go wrong, don't go with them! Switzerland
Disclaimer: "nobody in this company ever cared for what I said, why would they start now".
does anybody have some comments on or practical experience with the new Alexa fluorescent dyes from Molecular Probes?
Particularly, I would be interested in compatibility of Alexa488 and Alexa594 with normal Fitc or Texas Red filter sets, respectively. Do these dyes work in double labelings or is there a lot of bleed-through because they are so bright?
Thanks for your answers!
Peter Lorenz
Dr. Peter Lorenz University of Rostock Institute of Immunology Schillingallee 70 D-18055 Rostock Germany fax: +49 381 494 5882
Previous posts are true for larger particles which are good insulators. On the other hand, I often need to examine mounted and polished finely divided conductive and semi-conductive material. Quite often I am looking for hi-Z metal carbides. Carbon coating really hurts the desired to undesired/signal/noise ratio.
It is also true that conductor loaded insulating resins (bulk conducting) do not help much in this area. The islands of charging resin will "eat you alive" {g} .
To clarify my earlier post:
What I would like to see developed is intrinsically conductive polymer (sometimes know as unobtianium) that can be used to mount and polish specimen material (fines). This sort of polymer is currently available in thermoset (already) powder and, as another poster mentioned, solid forms (block, sheet, etc.). This polymer itself is conductive. Several different schemes may be used. One method, for example, adds an iodine atom in the polymer chain to free some conductance electrons.
Woody White McDermott Technology, Inc.
I agree with Jean Marc. All this excitement about conducting resins is not likely to achieve the aim, which was the elimination of charging in the SEM of DIFFICULT non-conductors embedded in resins. {snip}
Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** -----------------------------------------------------------------------.
Just a remark on conductive resins: they are good to embed conductive samples for surface SEM observation for example. But don't forget, they are composite materials, i.e. non-conductive resin loaded with enough conducting material (Cu, Ag whatever) to make them MACROSCOPICALLY conductive.
In microscopy application, their composite structure quickly appears and limit their usage. For example, their are useless to look at edges of conductive material because the non-conductive resin portion in-between the conductive particles will charge up exactly as pure polymer and blur out the picture.
For your ion-milling application I suspect the same holds true but I haven't
try myself. So just be aware of this problem if doesn't work.
BTW they are available from any electron microscopy accessories vendor.
Jean-Marc Boichat email: jean-marc.boechat-at-chma.mhs.ciba.com EM LABS FO 5.1 phone:+4126 435 6979 fax: +4126 435 6907 Ciba research Center P.O. Box 64 CH- 1723 Marly 1 When things go wrong, don't go with them! Switzerland
Disclaimer: "nobody in this company ever cared for what I said, why would they start now".
Could anyone tell me if there is a stain (or stains; vital or non-vital) that could help me differentiate between single-celled eucaryotes and procaryotes under the light microscope? Perhaps something that stains peptidoglycan, which most if not all bacteria have, but no eucaryotes have. Gram stain won't work since euks come up gram-negative. I'm trying to distinguish between nanoplankton and cyanobacteria growing on an inorganic media, without resorting to using a TEM (which we don't have!)
I work for an enginnering firm in New York City. My firm needs to obtain fracture surface images using a SEM. The pieces are made of rubber. The first piece is 1 3/4" in diameter and 1/4 inch thick. The second piece is 1 inche thick with a outside diameter of 1 3/4 inches and an inside diamter of 1 1/8 inches. We need to contract the use of an ESEM that is locate near NYC as we only have one day to scan the pieces. Please contact Carl Mallery (212) 972-7027 if you can help.
Interesting? What changes magnification in a SEM? Change the kV, workin= g distance, lens focal length and of course the angle of sweep of the scan coils, all change the magnification provide the instrument does not compensate the magnification for these changes.
Lets take a look at the optics. Increase the strength of C1 and this plac= es the first crossover higher in the column, which in turn places the second=
crossover higher in the column and similarly lifting the third crossover,=
the point where the final condenser places the probe on the surface of t= he specimen; the image goes "out of focus". Thus we have to decrease the strength of the final lens to bring the specimen back into focus =
Decreasing the final lens strength increases its focal length effectively=
increasing the distance between the pivot point of the scan and the specimen surface. Change the position of the pivot point of the scan without changing the scan angle and you change the length of the scan lin= e on the specimen, the magnification changes as magnification is the relationship between the length of the scan line on the specimen and the length of that same scan line on the CRT.
Try this test if you have a SEM that does not automatically compensate fo= r the focal length change in the software. Focus at one spot size, dramatically reduce the spot size (probe current) by increasing the strength of C1, the image will dim and go out of focus! Increase the gain/contrast and turn the third condenser anticlockwise (increasing the focal length), the image will come back into focus. I do not know the ra= te of change on every SEM but as an example the old Hitachi S520, S650 range=
with a ten turn spot size control changed the magnification by ~5% for every full turn of the potentiometer in the range 5 to 7 turns! =
Do this and you like I will have pictures to prove that a change in spot size in an uncompensated instrument does change the magnification!
Interesting?
Steve Chapman Senior Consultant E.M. Protrain, Oxford, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consulatncy and Courses in Electron Microscopy World Wide
We have a JEOL 840 SEM and are looking to fit it with a reasonably-priced EDS system. This is a workhorse instrument and the really detailed things like mapping or light element analysis would be done on another SEM (with a FEG) that has all the bells and whistles. Therefore we are just looking for something basic, user friendly and not too expensive. We have a Gatan DigiScan already in place.
Any comments from vendors would be welcome, as well as individuals who currently utilize a system that matches this general need, either on or off line. Thanks in advance.
Cheers,
JSV *************************** John S. Vetrano Sr. Research Scientist Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
---------------------- Forwarded by Rick Felten/MACDERMID/MACDERMID/US on 03/05/98 11:13 AM ---------------------------
Rick Felten 03/03/98 12:10 PM
To: Microscopy -at- Sparc5.Microscopy.Com cc:
Dear Friends:
I am thinking about converting my Cameca CAMEBAX microprobe from a turbo to a diff pump. Some people tell me that the diffusion pumped version is alot less troublesome than the turbo pumped version. Does anyone have any experience/recommendations/opinions about this conversion? Would anyone have a diffusion pump/vacuum controller (or an entire microprobe) they might be willing to part with?
Your assistance would be greatly appreciated.
Thanks in advance,
Michael Coviello EM Lab Manager Materials Science The University of Texas -at- Arlington Arlington, TX E-mail coviello-at-mae.uta.edu 817-272-5496
In reply to Trump's fixative: Solution of 4% formalin and 1% glutaraldehyde in 0.1M phosphate buffer, the original reference can be found:Arch Path Lab Med 100:405-414, 1973 They discuss the advantages of using Trump's for surgical pathology specimens for transmission electron microscopy. As far as immuno staining, a balance between preservation and antigenicity is difficult, the antigenicity is sometimes lost with glutaraldehyde, but this loss is antigen-dependent and some antigens will label even after fixation with glutaraldehyde. If I can be of further help, feel free to contact me Marge
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
Dear List, The last time this thread was on the List, I asked for a supplier of the German-made Technovit 5000, which is a copper-filled, cold-curing resin I have used in the past. The Canadian supplier was no longer carrying it. Energy Beam Sciences (www.ebsciences.com/) replied that they could get it on 5 to 7 weeks delivery. It is about $200US for 500 ml. liquid and 1000 g. copper powder, but this lasts a long time. The quality is O.K., with some resin areas that charge, but it is essential for looking at edges of material you don't want to carbon coat or heat up. Most metallurgical companies carry a conductive hot-press material.
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
I am thinking about converting my Cameca CAMEBAX microprobe from a turbo to a diff pump. Some people tell me that the diffusion pumped version is alot less troublesome than the turbo pumped version. Does anyone have any experience/recommendations/opinions about this conversion? Would anyone have a diffusion pump/vacuum controller (or an entire microprobe) they might be willing to part with?
Your assistance would be greatly appreciated.
Thanks in advance,
Michael Coviello EM Lab Manager Materials Science The University of Texas -at- Arlington Arlington, TX E-mail coviello-at-mae.uta.edu 817-272-5496
We have two disks in our standards collection which are marked as "C.M. Taylor" standards. One of these comprises mainly oxides, sulfides, etc.; for these we have analyses. The other - the map for which indicates that they are metals - we have no analyses for. Although this may, on the face of it, seem a silly question, what I need to know is if these are truly single metal standards - i.e., is the Pt really just Pt, the Co really just Co, etc.? For the lone analysis we have - that for the Hf metal - we see that it is actually a Hf-Zr alloy. Do any of you use these standards? Also, can any of you point me to a means of contact with C.M. Taylor Co., should they still exist that is.
Thanks, in advance, for time spent answering this.
Winton Cotnell
Dr. Winton Cornell Senior Research Associate Department of Geosciences 600 South College University of Tulsa Tulsa, OK 74104
The following is the text of a notice which will appear in next week's issue of New Scientist. I would be grateful if you could draw it to the attention of anyone who is job hunting and has suitable experience.
Many thanks, Mark Aindow, School of Metallurgy and Materials, University of Birmingham
**********************************************
THE UNIVERSITY OF BIRMINGHAM School of Metallurgy and Materials
Research Fellow
Microstructural Studies of High Tc Superconductors
Applications are invited for the above EPSRC-funded post which is available from April 1st for 3 years. The work will involve the use of transmission electron microscopy and other advanced characterisation techniques to obtain detailed measurements of the crystal structure, chemistry, morphology and defect content of high Tc superconducting oxides. This research will be part of the interdisciplinary activity in Birmingham which includes studies of bulk, thick film and epitaxial thin film materials. Applicants should have a PhD in a relevant subject area, a thorough understanding of crystallography and extensive experience of transmission electron microscopy.
The starting salary will be in the range...GBP15,159 - 21,894.
Preliminary enquiries should be directed to: Dr. M. Aindow Telephone: +44 121 414 5188, Email: M.Aindow-at-bham.ac.uk. or Dr. J.S. Abell Telephone: +44 121 414 5168, Email: J.S.Abell-at-bham.ac.uk.
Application forms (returnable by April 2nd) and further particulars are available from the Director of Staffing Services, The University of Birmingham, Edgbaston, Birmingham B15 2TT, Telephone +44 121 414 6483 (24 hours), (Email: Staffing-at-bham.ac.uk).
Please quote reference G9550/98.
Working towards equal opportunities.
************************************************
Mark Aindow, School of Metallurgy and Materials, Telephone; (0121) 414 5188 The University of Birmingham, FAX; (0121) 414 5232 Elms Road, Edgbaston, Birmingham, Email; M.AINDOW-at-BHAM.AC.UK GB B15 2TT, United Kingdom.
I have inherited an EDAX 9800 which is attached to an Amray 1830i. I was wondering if I could replace the 5.25 inch drive with a 5.25/3.5 inch drive? It makes sense to me, and would be practical, but the tech support at EDAX said that it would be impossible and crash the system; which does not make sense to me.....yet. Are they right at EDAX? Or, are they trying to get me to upgrade my system? Please enlighten me for I am an X-Ray novice.
John Grazul Rutgers University Electron Imaging Facility
C.M. Taylor Corporation still exists, their information is as follows:
Dr. Charles M. Taylor 289 Leota Avenue Sunnyvale, CA 94086 Phone: 408-245-4229 FAX: 408-526-9021
I hope this helps.
Laura L. Estok Asst. to the President M.E. Taylor Engineering, Inc. 21604 Gentry Lane Brookeville, MD 20833 Phone: 301-774-6246 =95 FAX: 301-774-6711 =95 e-mail: Metengr-at-aol.com
----There is no connection between M.E. Taylor Engr. and C.M. Taylor Corp= -----
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Steve,
I'll try your tests with our SEM as soon as time allows. I do always recommend internal standards with specimens whenever possible. One of the classics in biological work is catalase with a very well defined crystalline spacing combined with preps of virus and small protein molecules. Accurate measurements of particles adjacent to or resting over these lattices eliminate problems with hysterisis and all other potential situations which may effect magnification. I have rare need for such accuracy in SEM of biological material since most is done at fairly low magnifications. However, I would like to have a good internal standard for magnifications in the 1000-20,000 range. Others probably would benefit from standards for higher magnifications. Do you or anyone else have suggestions as to readily available material that could be used for this purpose? Something that is of very accurate size, conductive, in a dry or non-aqueous form, and easily added on to the surface of the specimen would be desirable. Debby Sherman
--------------------------------------
I use many different instruments each month and during courses this type of "problem" has to be discussed with the clients. Set out below are the tests we do to ensure we fully understand the magnification/focus system of the instrument we are using.
The first instrument that I used that compensated for a condenser lens change was the Cambridge 360, other models in the Cambridge/Leo range also do so. Most instruments have an error when changing kV. The best check is to set a particular WD if this reads out? Having done this change the kV and see if the WD readout changes. As you did not move the specimen any change in WD indicates an error in the system as it compensates for the kV change. Of course a different WD readout will almost certainly mean a magnification "difference" between the two kV being investigated.
Test 1
Change the spot size/ probe current and if the instrument compensates the image will stay approximately in focus. If you are not sure, then the best check is to judge the correction in focus. If the system does not compensate, the focus correction for smaller and smaller spots will always be in the same direction, and the converse for bigger spots. If the system tries to compensate it often fails to get it absolutely correct so the new focus will be erratic rather than in a constant direction. This is a curse at high magnification as the extra time required to try and find focus results in higher levels of contamination; a constant error means a constant correction!
Test 2
Change the third condenser/focus control and watch the magnification display, it too should change in a "magnification compensation" system. There are systems that compensate the third lens/magnification readout but do not automatically compensate the image focus.
Test 3
Take magnification calibration pictures using a wide range of spot sizes/probe currents and measure the results.
I worry very much about people placing too much credibility on SEM magnification. There are so many things that may go wrong when judging a magnification that I like to think of it being ABSOLUTELY ACCURATE plus minus A FOOT! Flat specimens fine, but how many people with vastly undulating specimens use the Z or WD to compensate for a focus change? Changing the final lens, due to magnetic history, will change the focal length with a degree of inconsistency and therefore change the magnification. Have you tried doing stereo pairs and have you experienced the grief that the magnification changes can bring?
Best wishes and happy scanning.
Steve Chapman
Senior Consultant E.M. Protrain, Oxford, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consulatncy and Courses in Electron Microscopy World Wide
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Peter,
I have tried, and now use routinely, Molecular Probes Alexa 488 secondary antibodies as well as Alexa 568 Phalloidin. Both label brightly with no bleed through using standard Nikon filter cubes. I use the phalloidin at a 1:200 dilution of the methanolic stock directly into my working stock.
Steve Samuelsson ______________________________ Reply Separator _________________________________
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Hi,
does anybody have some comments on or practical experience with the new Alexa fluorescent dyes from Molecular Probes?
Particularly, I would be interested in compatibility of Alexa488 and Alexa594 with normal Fitc or Texas Red filter sets, respectively. Do these dyes work in double labelings or is there a lot of bleed-through because they are so bright?
Thanks for your answers!
Peter Lorenz
Dr. Peter Lorenz University of Rostock Institute of Immunology Schillingallee 70 D-18055 Rostock Germany fax: +49 381 494 5882
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If it is a standard Windows, PC-based system, I see no reason why you could not do that. If your floppy is currently A:, your hard drive is C:, and you have nothing defined as B:, you should be able to do it straightaway. That is the reason for going with standard hardware platforms and operating systems.
There is a possibility that the operating environment might be a bit unusual if it is an old PC. It would be more likely that EDAX had somehow customized it or used a non-standard system. It might also be a little tough to get to the CMOS setup program to inform the system what the new hardware configuration is, but you should be able to do it. EDAX may simply be trying to protect itself.
Things are much better now than in the old days when expansion was just about impossible. The EDS manufacturers did well with what they had. They sometimes made their own controller cards or wrote their own operating system (I think of my old TN-2000) because standardized, cheap components were not available or because they needed more performance than the standard parts could provide. Matters got better as the PDPs matured and RT-11 and TSX developed as the operating systems - then you could add your own components. But now it is just about a no-brainer. Indeed, I would rather avoid the proprietary systems. They might have a bit of technological edge today, but what will I do tomorrow when I want to slip in a faster processor or a 10 GB disk drive and tape backup?
At 08:24 AM 3/6/98 EDT, you wrote: } X-Rayers, } } I have inherited an EDAX 9800 which is attached to an Amray 1830i. I } was wondering if I could replace the 5.25 inch drive with a 5.25/3.5 } inch drive? It makes sense to me, and would be practical, but the } tech support at EDAX said that it would be impossible and crash the } system; which does not make sense to me.....yet. Are they right at } EDAX? Or, are they trying to get me to upgrade my system? Please } enlighten me for I am an X-Ray novice. } } } John Grazul } Rutgers University } Electron Imaging Facility
Since upgrading from the original Snappy to the Snappy 3, we have problems with acquiring. I wonder if others are using Snappy and if they are experiencing similar problems.
We upgraded to Snappy 3 for the enhanced resolution. When it works, it works very well. Our problem appears on frame averaging. Instead of taking the multiple images and averaging or interpolating them, we see two separate smaller images in the upper right and left corners above one large image which extends from the left and right edges. This appears to be the same picture shown on the screen in three separate frames.
I placed a call into Snappy when this first happened and they claimed they never heard of this and asked us to do a fair amount of programming and photographic documentation at various stages. While I would like to assist them (and myself), we just don't have time to troubleshoot their product to the extent they asked. If necessary, I will simply return the product to computer store where we bought it and go back to the old Snappy unit.
Has anyone out there had this problem? If so, how was it resolved?
Dear Phil & Mark, } } } A colleague needs to know the mean free path of 100keV electrons in ZrO2. } } He is trying to determine the absolute thickness of samples examined by } } PEELS. The EL/P program calculated a relative thickness in terms of the } } number of mean free paths but we want to convert this to thickness in } } nanometres. Any help would be appreciated. cheers, } } For a handy rule of thumb, consider assuming that } the inelastic mfp in most things is about 25 ug/cm^2 } for 100kV electrons (that's in "micrograms per square } centimeter"), and about 50 ug/cm^2 for 300kV electrons. } Dividing by the density of your ZrO2 would then give } you a 1st-order estimate of its thickness. The } EELS experts in this forum can probably provide a } more precise value for this in your case, as well } as information on its (I think relatively weak) } dependence on atomic number. } The mean-free-path is inversely proportional to the stopping power of the material, and the stopping power is proportional to the number of atomic (as opposed to beam) electrons per unit volume. The stopping power is -dE/dx, where E is the energy of the incident particle and x is thickness. There are small corrections for the chemical form of the material, but the stopping power for a compound or homogeneous mixture is essentially the sum of the stopping powers of each of the constituent elements. Since low-Z atoms have the same number of neutrons as protons, i.e. A = 2Z, and since the mass is essentially A, light elements have more electrons per unit mass, so their stopping powers are greater and the mean-free-path shorter than for higher-Z materials, where 1 { A { ~1.5--this is the relatively weak Z-dependence referred to. Expressing stopping power in units of cm^2/gm removes the factor of the density and leads to the near uniformity of the stopping power and near equality of mean-free-path. Yours, Bill Tivol
You did not mention computer type or OpSys... Cannot speak directly to problem, but here is possibility.
If the system uses non-PC/DOS/Win like many older x-ray systems, what ever is serving as a disk drive controller/BIOS may not "know" about the formatting requirements for a 3.5" drive.
Woody White McDermott Technology, Inc.
X-Rayers,
I have inherited an EDAX 9800 which is attached to an Amray 1830i. I was wondering if I could replace the 5.25 inch drive with a 5.25/3.5 inch drive? It makes sense to me, and would be practical, but the tech support at EDAX said that it would be impossible and crash the system; which does not make sense to me.....yet. Are they right at EDAX? Or, are they trying to get me to upgrade my system? Please enlighten me for I am an X-Ray novice.
John Grazul Rutgers University Electron Imaging Facility
At 09:13 PM 3/2/98 -0500, Dennis P Smith wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The new book, "Optmizing Light Microscopy for Biological and Clinical Laboratories" is a great reference. It begins with alignment, reviews not only the basic techniques and some of the advanced ones but a bit of the optical principles behind them so that you can use them to best advantage, and has lots of quick and easy experiments which you can do at your own microscope to get started. It is available directly from MME or from the American Society of Clinical Laboratory Sciences or from Kendall Hunt.
If you would like an order form from us, please email me privately.
Good hunting!
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street Springfield, MA 01118 (413)746-6931 FX: (413)746-9311 email: mme-at-map.com -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- -at--at--at--at- Microscopy/Microscopy Education: America's first national consortium of microscopy experts offering customized on-site training in all areas of microscopy, sample prep, and image analysis. Our goal is to help you optimize your microscopy. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- -at--at--at--at-
I would appreciate an answer to the following query from anyone attending/organizing ICEM-14 at Cancun.
The hotel reservation form asks us to write down the $ amount to be charged to guarantee the room reservation. Does this mean that we have to prepay the entire cost while making the hotel reservation?
Sarath K Menon Department of Mechanical Engineering Naval Postgraduate School Monterey, CA 93943
POSTDOCTORAL FELLOWSHIP OR GRADUATE STUDENT INTERNSHIP IN ANALYTICAL ELECTRON MICROSCOPY at the Federal Energy Technology Center, Pittsburgh, Pennsylvania
One postdoctoral fellowship or graduate student internship is available immediately for the development and application of SEM/EDS in the microstructural characterization of ash deposits generated from the firing of pulverized coal, biomass, and mixtures of these fuels.
Duties and Responsibilities include development of sample preparation techniques, SEM/EDS operation, implementation of SEM/EDS automation, and image analysis. The successful candidate should be prepared to play a central role in the interpretation and application of results in the context of a multi-disciplinary, multi-laboratory effort to develop new ash management tools.
Qualifications require experience in SEM/EDS operation, SEM/EDS automation, image analysis, and sample preparation---with ash, or similar materials.
The fellowship/internship appointment is for one year, with possible renewal. The Oak Ridge Institute for Science and Education (ORISE) will administer the position, and the salary is very attractive. U.S. citizenship or permanent resident alien status is required.
For further information contact:
Dr. Everett Ramer Combustion and Cleanup Division Federal Energy Technology Center P.O. Box 10940 Pittsburgh, PA 15236-0940 Phone: (412)892-4920 FAX: (412)892-4152 E-mail: ramer-at-fetc.doe.gov
Related presentations and reports are available via FTP at Titan.petc.doe.gov/pub/ramer/docs
Hi, This is a "case of emergency". I would like to do Quick freeze-deep etch technique in our lab to study the shape of a protein we purified. We have everything to start except the copper block that would allow me to freeze my samples. We have a slammer designed by Tom Heavy.I am looking for a copper block. I am planning to use nitrogen to do so. So, if you have an extra copper block to lend or to give or sell. Please, tell me.
In response to some comments that I have received about changing from a turbo to a diff pump. I would like to explain that some people believe that on the Cameca CAMEBAX microprobe, the turbo and the related controllers are troublesome and one would have less problems if one "retrogrades" to the simpler (and less troublesome) diff pump.
Thank you for all of your comments so far--please keep them coming.
Regards,
Michael Coviello EM Lab Manager Materials Science Dept. The University of Texas -at- Arlington Arlington, TX E-mail coviello-at-mae.uta.edu 817-272-5496
My daughter prepares a thesis on artistic paintings and would like to know if there are recent publications on the use of the SEM and EDX on restoration of painting, expertise or research in history of art.
To test if you can connect to SATNET, put in the subject your "ECUSA subscription number", or put "DELETE" ( to delete your name from SATNET files ), then click REPLY.
I am faced with the problem of counting viral particles of human parainfluenza virus type 3 with the negative staining technique of 50ul sample. Personaly I do not know how to do it and I am not sure if it is possible to do? I need some advise from a person who is more familiar in this field. Any advice will be appreciated. TIA John Gabrovsek CCF Cleveland, Ohio
I would like to identify anyone in the Houston, Texas, USA area with Hi-Res TEM capability, willing to image for fee or sell beam time. This request is for both immediate service & to identify local EM resources. Please contact me directly.
} Date: Fri, 06 Mar 1998 17:02:43 -0500 } From: John Gabrovsek {GABROVJ-at-cesmtp.ccf.org} } To: MICROSCOPY-at-sparc5.microscopy.com } Subject: TEM virus count } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am faced with the problem of counting viral particles of human } parainfluenza virus type 3 with the negative staining technique of } 50ul sample. Personaly I do not know how to do it and I am not sure if it is } possible to do? I need some advise from a person who is more familiar in } this field. Any advice will be appreciated. } TIA } John Gabrovsek } CCF } Cleveland, Ohio
You will have a very difficult time enumerating these viruses because they are enveloped, come in different sizes, may stick to cells and pellet out with cell debris, some may not take the stain in such a way as to show the spikes clearly so that they can be differentiated from cell debris, and some may stick together in a pile so that you can't tell how many are there. Counting viruses is easier with icosahedral viruses.
Nonetheless, if you can live with a rough estimate, you can use a Beckman Airfuge and pellet the virions onto a Formvar and carbon-coated grid. You will have to work out a dilution so that you have a countable number (hundreds, not thousands). You should do several grids and count several areas on the grid square. We count 3 different areas in a triangle on the grid and 3 grid holes in each area. If the numbers are similar (same log), we accept it; if numbers vary widely, we repeat.
We sonicate our preps in a water bath sonicator before pelleting. What will this do to your enveloped virus??? Also use glow discharged grids. Make sure you've recentlly calibrated the mag on your microscope, and calculate the area in a grid square. Count the viruses at a high enough mag so that you can see that they are really viruses (not just cell debris)--something like 60-80 KX, and go back and forth up the corn rows, trying not to recount the same area. If you take pictures at a low mag and then project them onto a wall to count, make sure the resolution is such that you can tell that the blobs are really viruses, and take lots of pix.
Good luck!!!
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
What dilution of the Alexa 488 secondary antibody concentration do you use? I plan on trying it next week and Molecular Probes suggested that I start at a dilution of 1:2000. Thanks.
Patty Jansma Tel:520-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
On Fri, 6 Mar 1998 samuelsson.sj-at-pg.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Peter, } } I have tried, and now use routinely, Molecular Probes Alexa 488 secondary } antibodies as well as Alexa 568 Phalloidin. Both label brightly with no bleed } through using standard Nikon filter cubes. I use the phalloidin at a 1:200 } dilution of the methanolic stock directly into my working stock. } } Steve Samuelsson } ______________________________ Reply Separator _________________________________ } Subject: LM- Alexa dyes } Author: (INTERNET)peter.lorenz-at-med.uni-rostock.de at external } Date: 3/5/98 2:45 PM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi, } } does anybody have some comments on or practical experience with } the new Alexa fluorescent dyes from Molecular Probes? } } Particularly, I would be interested in compatibility of } Alexa488 and Alexa594 with normal Fitc or Texas Red filter sets, } respectively. Do these dyes work in double labelings or is there } a lot of bleed-through because they are so bright? } } Thanks for your answers! } } } Peter Lorenz } } } Dr. Peter Lorenz } University of Rostock } Institute of Immunology } Schillingallee 70 } D-18055 Rostock } Germany } fax: +49 381 494 5882 }
Spi TEM Crossed Carbon Grating Replicas are the magnification standards that we use on courses. Although this is a TEM specimen it makes a very interesting general SEM standard as it may be used for magification calibrations and if mounted carbon side up as a kV guide. In the latter case the carbon film at low magnification is only visible below 5kV depending upon the microscope geometry. You may have seen these results,=
often credited to David Joy, although we published in 1986!
These specimens are only 3mm across and if tacked to a small thin alumini= um disk they will last a long time being stuck and unstuck to a conventional=
specimen stub. We use 2160 lines per mm or 0.4629 micron grating space.
The only problem with the specimen is that you do need a good degree of S= EM operating skill to visualise the very thin carbon film. The magnificatio= n range covered is easily 5,000X to 40,000X.
Regards
Steve Chapman
Senior Consultant E.M. Protrain, Oxford, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Regarding the recent discussion on conducting a new salary survey: Great idea!
The USA 1996 wage tables are now available at http://stats.bls.gov/oeshome.htm which provides listings of average and median wages at the national and state level. Microscopists don't rate a category. Find yourself under one of the scientist or technologist categories.
I've also recently seen a well-done salary survey in an electrical eng publication... could have been ieee (?) ... ideas worth copying.
Thank you to my fellow microscopist who suggested painting the inside of the video camera adapter with graphite. I tried this and it cut down the glare substantially. I still have a little when the illumination is set high, but this is a great improvement.
Alan Stone Alan Stone ASTON Metallurgical Services Chicago
However it does have a focal length that may be varied and it is the change of focal length varying the "effective" working distance that ultimately changes the magnification.
Try the tests I mentioned in an earlier submission and you will see for yourself.
I was originally a TEM engineer, with all the tests in mind that we did 30 odd years ago to prove our TEM, I have over the past 20 years applied the TEM monitoring techniques to the SEM. Such data as reasons for magnification changes, contamination rate tests and resolution tests with different standards are gathered on almost a monthly basis through certain of the courses that we run.
If these tests were carried out in all SEM laboratories the truth about magnification and the features that cause it to change would be readily available. Do we train correctly if we produce operators ignorant about magnification errors, or do we not care about such detail; its probably a bit of both!
Steve Chapman
Senior Consultant E.M. Protrain, Oxford, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Group, Thanks to the help from a number of you, I am about ready to launch the microscopist salary survey. If done properly, I expect that the results may be of real value to some of you, and as my approach is a bit different than many other surveys I have considered, I would like to offer this (last) preview as to format and invite any comment. You will be invited to provide your information via this listserver. If so, after recording your data I will dump your message. I have NO interest in your individual information. However, if you wish you can provide your data on a completely "blind" reader response card in our next (April) issue of Microscopy Today. The survey will be initially open only to microscopists in the U.S. and manufacturers and suppliers will be excluded. First, we will plot salary level against years of microscopy experience (in 3 year increments) for educational levels (no degree, AA, BS, MS, PhD and MD/DVm) Then, there will be only four criteria as follows: 1) Gender - Male/Female 2) Geographical Location - Midwest, West, South, Southeast, Northeast. If in question, one should pick the area which he/she feels most accurately reflects their salary level. For example, one from Tucson may feel their salary level to be closer to L.A./San Francisco than Houston/Dallas - so would pick West. 3) Primary area of interest: Biological, Physical/Material or Earth Science. 4) Working in: Industry, Education, Government, or Medical
Then for each criteria entry we will compute a "factor" based upon the average salary for all microscopists. For example, say that the average salary for all microscopists was $30,000 - and the average for all females was 2.3 % lower. The the female salary factor would be minus .023. Or say that the average salary for microscopists working in biology was 2.8 % higher than the total salary average - then the factor would be plus .028
Then, one could determine their base salary level from the educational/time curve and apply the four appropriate factors to come up with their own approximate salary level.
I have, of course, considered other criteria, like working with electron, optical, etc. microscopes, but do not see any real differences that would effect salary (?)
Sure, this is not absolute, but is (I submit) more accurate than anything else that I have been able to come up with.
I would appreciate any comment any of you might have as to how to improve on the system. I would like to get the survey started in a weeks time. Regards, Don Grimes P.S. If you "like" this and have no constructive comments, kindly reply direct to me as to not clutter up the system. Thanks
San Francisco Microscopical Society 12 March 1998 6:30 PM
Technical Instrument Company 348 Sixth Street San Francisco, CA 94103
Topics: The Antique Microscope Collection of Technical Instrument Co. Sale of Used Microscopy Equipment
You will not want to miss this unique opportunity to inspect the extensive antique microscope collection of Technical Instrument Company. In addition, Technical Instruments will have used equipment for sale and there will be a silent auction with proceeds to benefit the Society. This is your opportunity to find that microscope, accessory, or gizmo that is just what you need to do what you've always wanted to do. This should make for a wonderful evening in The City. Please join us!
Further Information: Contact Peter Barnett, Forensic Science Associates, 510-222-8883 or see the *relocated* SFMS Home Page at:
http://www.microdataware.com/sfms/index.htm
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and in limited service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
} I am trying to gain some understanding of the workings of TEM. Does anyone } know of any URL's that contains some good introductory material for TEM. } Kelvin,
try
http://biosci.cbs.umn.edu/biophys/OLTB/diffract.html It is part of the "Online Biophysics Textbook" and there's a pdf file by Wah Chiu et. al on EM
There's a lecture on EM by Aebi et al. in http://www.mih.unibas.ch/Booklet/Overview.html
You could search through "Frank Potters's Science Gems": http://www-sci.lib.uci.edu/SEP/SEP.html
Hope that helps a bit,
Philip -- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
Within the frame of a European research project on coatings for Ultra violet applications, several post-doc positions are available, two of them related to TEM techniques. The applicants should be members of the EU. The host center for this two ones is University of Barcelona and the main objectives of the work is the structural and compositional characterization of UV-coatings.
Is someone is interested, please contact Dr. F. Peir=F3 at
paqui-at-iris1.fae.ub.es
Please find more information visiting the web site: www.lzh.de/tmr/default.htm
*******************************+ Francesca Peiro
EME, Enginyeria i Materials Electronics Dpt. Electronica Universitat de Barcelona Avda. Diagonal 645-647 08028 Barcelona, Spain
Greetings All ... I was wondering where everyone keeps their stock of copper grids. My predecessor always kept them in the film desiccator in the JEOL , JEM-100CX II. This was to keep the copper from excess moisture and prevent premature corrosion. I still do that, but what is everyone else doing?
Sharron G. Chism HT (ASCP) Electron Microscopy Department Harris Methodist Fort Worth Fort Worth, Texas
} Hi all I am a novice of this list, so please write me is there is some FAQ to avoid starting again old threads. } By the way, is there a consolidated way of protecting a SEM chamber from boost pressure coming from N2 bottle during venting? } (this may occur if someone accidentally moves the low pressure stage on the bottle) } } Thank you in advance } Dr. Eng. Edoardo Bemporad, Ph. D. } Assistant Professor of Materials Science } University of Rome "Roma Tre" (Italy) } Dipartimento di Ingegneria Meccanica e Industriale } (Department of Mechanical and Industrial Engineering) } Via della Vasca Navale 79 - 00146 Rome, Italy } Tel: +39 6 5517.3293 } Fax: +39 6 5517.3256 } LIME Lab (InterDipartimental Laboratory of Electron Microscopy) Tel: +39 6 5517.3200 } E-Mail:bemporad-at-uniroma3.it
Overpressure in SEM chambers is only a problem on some SEMs with airlocks for sample loading. For direct loading SEMs, the stage door opens when the chamber is above atomospheric if you don't fasten the door down. On JEOL SEMs there is an overpressure relief valve on the N2 Backfill line and some other SEMs also have an overpressure valve.
If you often vent the specimen chamber, just make sure the stage door opens freely or that a port flange can pop open. Just loosen the port flange screws. Atmospheric pressure holds these tight against the vacuum so there is no need need for tightness except at the beginning of pump down.
At 10:28 PM 2/25/98, Gabriel Adriano Rosa wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just saw their representatives last week at PittCon. Please specify needed information.
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite B Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
I have to examine some gunk from an industrial fermentation + processing reaction. I'd appreciate any suggestions for an accessible reference on reagents for microchemical tests to distinguish protein, carbohydrate, cell wall materials, or other broad classes of chemical materials, mostly organic or bioorganic, though there may also be inorganic components.
Leonard Corwin Fort Dodge Animal Health (Analytical Research) Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
The 9800 has an RT-11 based DEC computer and it was a momentous occasion just going from 8" to 5.25" drives! As far as we know, DEC never developed the driver needed to control a 3.5" floppy on this system. (If one does exist, the next issue would be compatibility with the high density drives you get off the shelf these days.)
As you inherited the system and are new to EDS, you might be interested in knowing the "i" in 1830i stands for "integrated". Amray purchased 9800 analyzers from EDAX and used them for microscope control, too. These integrated systems were sold and serviced by Amray, so we might not even know where your system was prior to reaching your lab. This also means our tech support people have to exercise some extra caution.
Just so you know, you can get our latest and greatest PC software to process your data without upgrading your hardware. Files can be captured with Kermit over a serial connection, but most people opt for the sneaker net route. If you have a PC with a 5.25" drive, our utilities transfer and convert the data from RT-11 floppies to the DOS/Windows world. As 5.25" drives aren't as common as they used to be, I'd love to be able to retrofit 9800's with 3.5" drives. Of course, if we make it too easy for these 10-15 year old machines to keep going, I won't sell any upgrades ;-)
Hope this helped and welcome to EDAX. Be sure to let us know if there is anything we can do to help.
Jeff Gschwend Midwest Sales EDAX Inc.
Once upon a time, John Grazul shaped the electrons to say:
} X-Rayers, } } I have inherited an EDAX 9800 which is attached to an Amray 1830i. I } was wondering if I could replace the 5.25 inch drive with a 5.25/3.5 } inch drive? It makes sense to me, and would be practical, but the } tech support at EDAX said that it would be impossible and crash the } system; which does not make sense to me.....yet. Are they right at } EDAX? Or, are they trying to get me to upgrade my system? Please } enlighten me for I am an X-Ray novice. } } } John Grazul } Rutgers University } Electron Imaging Facility
At 04:44 PM 3/9/98 -0400, corwinl-at-pt.cyanamid.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Re: a good resource on microchemical tests The best one which I have seen is Chamot & Mason, Vols I & II. They were out of print for some time, but I saw them offered through McCrone Associates not too long ago.
Re: the organic goop The best solution, here, is an FT-IR microscope. There are a number of companies offering them, ranging from Perkin-Elmer to Nicolet/Spectra-Tech. I'd suggest referring to the American Lab Buyers' Guide for a listing.
For tips on the difference between sample prep for conventional vs. FT-IR microscopy, try the article I co-authored with John Reffner: Foster, B. and Reffner, J. "Focus on Microscopy: Conventional Optical Microscopy vs. FTIR Microscopy: Sample Preparation for Both Sides of the Coin". Part I: Sept. 1995, 16c-16J Part II: Nov. 1995, 20i-20M (We have reprints here, if you need one).
Good hunting! Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite B Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
In regard to Steve Chapman's comments about SEM magnification changing with condenser setting: I am not in a position to disagree with his statement regarding what he observes happening to the magnification when the condenser lens strength is altered. However, I disagree with the argument advanced to support why this happens. Quoting from his earlier posting:
} Lets take a look at the optics. Increase the strength of C1 and this places } the first crossover higher in the column, which in turn places the second } crossover higher in the column and similarly lifting the third crossover, } the point where the final condenser places the probe on the surface of the } specimen; the image goes "out of focus". Thus we have to decrease the } strength of the final lens to bring the specimen back into focus } Decreasing the final lens strength increases its focal length effectively } increasing the distance between the pivot point of the scan and the } specimen surface. Change the position of the pivot point of the scan } without changing the scan angle and you change the length of the scan line } on the specimen, the magnification changes as magnification is the } relationship between the length of the scan line on the specimen and the } length of that same scan line on the CRT. } I agree with all of this statement until we get to the part about the pivot point of the scan being affected by the strength of the final lens. The scan coil system should be designed such that the pivot point of the scan is located at the principal plane of the final lens where it will be unaffected by the strength of the final lens. This is simply an application of basic geometric optics: any ray which passes through the "center" of a thin lens (ie. the point where the axis intersects the principal plane) is undeflected. (That's why this undeflected ray is universally used as one of the "cardinal rays" in a lens diagram.) Now if the scan system pivot point is located substantially above or below this "center" position, then the final lens strength will indeed affect it and there will be an observable magnification effect. I would personally consider this to be a malfunction of the scan coil system. Though the location of the pivot point is unlikely to be exactly centered in the principal plane of the final lens, a small error introduces only a small effect and (assuming a properly functioning unit) I doubt whether this could explain the effect that Steve is discussing. A large error would also manifest itself in a change in apparent size of objects as the final lens strength is adjusted. In a properly operating SEM optical system, one should be able to change the strength of the final lens and although the image of a specimen feature will go out of focus and rotate, it should not change noticeably in size. This is, in fact, a simple way to check whether the scan pivot point is located where it is supposed to be. (For example, if the x and y scan coil sets pivot at substantially different points, this exercise will result in an apparent elongation of the image as one goes far out of focus.)
One possible point of contention is whether the principal plane of the final lens is indeed fixed. The answer is that for a highly assymetric lens (such as is typical of the SEM final lens) the principal plane does move slightly with differing excitation. However, it is my experience that this shift is relatively small (something that the lens designer should be concerned about) and is in fact too small to effect a noticeable (ie. first order) change in magnification.
As I stated above, I am not in a position to argue that the apparent magnification of some microscopes does not depend on the condenser setting. I would argue, however, that this is an artifact of the implementation, since I do not believe it is intrinsic to the SEM optical system One question to ask is whether, when it is stated that the "magnfication" changes, are we actually refering to the size of objects on the viewing screen, or to the numeric "magnfication" readout of the scope? My contention is that the former should NOT change in any substantial way. But it is easy to see how the latter might change in the way described. As Steve has pointed out, magnification depends upon both the rocking angle of the scan and the distance to the specimen surface. The latter distance, however, must be inferred somehow and typically this is accomplished in the microscope's readout circuitry via a calibration of working distance versus final lens excitation. Thus, a large change in condenser strength will (at least in some optical systems) require a substantial compensation in final lens strength which will throw off the working distance calibration and thus introduce an error into the magnification readout. This mechanism, however, results in a change only in the magnification READOUT of the microscope and the apparent size of objects on the viewing screen should not be affected. Could this be the nature of the "magnification" change being described? -- Fred Schamber RJ Lee Instruments Limited
INTERNATIONAL COURSES OF LIGHT MICROSCOPY, PHOTOMICROGRAPHY AND LASER SCANNING CONFOCAL MICROSCOPY GARGNANO (Lake of Garda) October 1998
The Course is a post-graduated theoretical/practical course, with propedeutical lectures and practical stages on microscopy, photomicrography and confocal microscopy. The course will take place in Gargnano (Lake of Garda) in October 1998.
All information and registration details (participation fee, date, special accomodation) at at the following Web address.
http://imiucca.csi.unimi.it/endomi/micro.html
Thank you Paolo Castano
_____________________________________________________ Prof. Paolo Castano UNIVERSITY OF MILAN INSTITUTE OF HUMAN ANATOMY - CHAIR OF HUMAN ANATOMY FOR PHARMACY Via Mangiagalli, 31 - 20133 Milan (Italy)
Hi Sharron, I keep my grids in the orginal vials, separated in a plastic divided storage box in a drawer of my microtome. Nothing special. When I go to use them, I do dip them in a 0.25% solution of formvar before picking up grids.
Best of luck, Ed Calomeni Medical College of Ohio Dept. Pathology Toledo, OH 43614 ecalomeni-at-mco.edu
All: I'm looking for labs to do material science (semiconductor) SEM/EDX/TEM/FIB work. I would prefer to hear about labs in the Dallas/Fort Worth or Texas state area but will take anyone in the US.
Does anyone know a site to downlad a free software for 3D reconstruction easy to manage? I need to reconstruct an anatomy sample by means of serial sections.=20 I=B4l intend to take microphotographs (LM level) of these sections, scan= them and finallly digitalize these images in tiff format. Is this procedure= correct? Thanks, in advances. Biol. Luis Ernesto Dettin dettin-at-cmefcm.uncor.edu Centro de Microscopia Electronica Facultad de Ciencias Medicas Universidad Nacional de Cordoba Te/Fax: 01-051-333021 E-mail: dettin-at-cmefcm.uncor.edu CC 362 5000 Cordoba Argentina
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RE} LM: microchemical class reagents 3/10/98 9:12 AM Dear Leonard,
You can, of course, try the Chamot and Mason books. However, the better r= eferences are by Feigel, "Spot tests for Organic and Inorganic Analysis".= These are spot test methods for small abounts of chemicals/materials. Th= e Feigel books are unsurpassed as references for microchemical tests.
Give us a call if you need further information or if we can help. We also= teach classes in microchemical testing. See our website.
John Shane Director of Research McCrone Research Insitute 2820 South Michigan Ave. Chicago, IL 60616
I have to examine some gunk from an industrial fermentation +=20 processing reaction. I'd appreciate any suggestions for an accessibl= e=20 reference on reagents for microchemical tests to distinguish protein= ,=20 carbohydrate, cell wall materials, or other broad classes of chemica= l=20 materials, mostly organic or bioorganic, though there may also be=20=
inorganic components. =20 =20 =20 Leonard Corwin Fort Dodge Animal Health (Analytical Research) Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
via sendmail with P:smtp/R:bind_hosts/T:inet_zone_bind_smtp (sender: {yoyodine-at-unm.edu} ) id {m0yCT0q-0001BRC-at-lyra.unm.edu} for {Microscopy-at-Sparc5.Microscopy.Com} ; Tue, 10 Mar 1998 10:40:24 -0700 (MST) (Smail-3.2.0.101 1997-Dec-17 #6 built 1998-Jan-5)
Hello,
I am looking for John Armstrongs CITZAF program. We have down loaded one from Argon NL's ftp site but the Self Extracting file CITPTC.exe won't self extract. Does anyone know a different source for the programm?? It is for a PC.
Mi chiamo Corneliu Mateescu PhD, con una esperienza di quasi 30 ani in ricerche sulla biologia di cellula cancerogena. Io sono capo di Dipartimento di Citometria e Microanalisi d'imagini microscopiche. Da 15 ani ho iniziato e svilupato un "soft" di analissi d'imagini per anatomia patologica e citopatologia neoplastica. Adesso il mio "soft" non corisponde alle esigenze di INTERNET, ma ci sono qualche elementi pregiatti. Il mio Consilio Scientifico e io personalmente sono molto interesato in una colaborazione di duratura con un partner straniero per iniziare una base datti per imagini microscopici. Abbiamo un "tezoro" di decine di milliai di lastrine chi aspettano solo una metodologia su la surveglianza internazionale d'iniziare questo base datti. Quale e la vostra oppinione ? Certo d'una Vostra solecita risposta vi porgo i miei mugliori saluti,
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Im sorry if this is a REALLLLL Dumn question.......... But just what is the } size of some of these TEM,SEM ect } Ron Mayer } thetruth-at-southwind.net } } Point your browser to eps.unm.edu/probelab . There are some good digital images of some typical machines there. SEM's TEM's etc....do vary in size though.
Greetings, I would like to ask for suggestions on fixation of cultured, embryonic frog muscle cells and the retention of good exterior membrane morphology. We grow Xenopus muscle somites (frog stage 20-23) in culture for 24-48hrs for study of membrane acetylcholine receptor aggregation with FESEM. The mucsle cells are undifferentiated at 24 hrs and retain a round shape rathar than the bipolar flat muscle cell morphology. The cells are not well developed at 24hrs and have a central cytoplasm filled with yolk granules and a clear non-structured peripheral cyotplasmic region. After aldehyde and osmium fixation the membrane (viewed in FESEM at 50 to 100K) has tiny holes and a shredded-torn appearance. We have developed a fixation regime that seems to produce very little cell shrinkage (less than 10%) and cell distortion (membrane ruffling) as viewed in the light microscope. However with the FESEM our membrane surface has the problems described. I have listed below our complete fixation protocol and would appreciate any suggestions.
Thanks in advance, Dennis Kunkel
Protocol:
1. Muscle somites are grown on glass coverslips for 24 hrs in culture medium.
2. Aldehyde fixation - 4 step After removing most of the medium the first fixative is added:
A. 1% paraformaldehyde in dH20 containing 0.15M sucrose and 10mM Ca and Mg - 30 minutes. *Note the dH20 is used - testing of many buffers in combination with sucrose to protect the cells osmotically did not work; the cells responded best with dH20.
B. 2% paraformaldehyde in dH20 containing 0.15M sucrose and 10mM Ca and Mg - 30 minutes.
C. 2% paraformaldehyde and 1% glutaraldehyde in dH20 containing 0.15M sucrose and 10mM Ca and Mg - 30 minutes.
D. 1% paraformaldehyde and 2.5 % glutaraldehyde in dH20 containing 0.15M sucrose and 10mM Ca and Mg - 30 minutes.
3. Rinse in 0.1M sodium cacodylate.
4. Fix in 1% osmium in 0.1M sodium cacodylate - 30 mins
5. Rinse in 0.1M sodium cacodylate
6. Dehydrate in 10% graded ethanol series to 100%
7. CPD using CO2 critical point dryer.
8. Cells are coated with gold/palladium to check membrane surface morphology. For other high resolution observations, using antibody and secondary gold labeling, cells are coated with carbon.
*********************************************** * Dennis Kunkel Ph.D. * * Pacific Biomedical Research Center * * University of Hawaii * * * * email - kunkel-at-pbrc.hawaii.edu * ***********************************************
We have a Leica-Wild M650 micro-surgical stereomicroscope (purchased originally in 1992) that we would like to trade/exchange for a Wild M420 with macro ApoZoom. Please contact me directly if you are interested.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Reference our discussions in this area, as we would say "you hit the nail=
on the head". =
A change in final lens strength confuses the calculation of magnification=
(angle verses WD) and thus the magnification calibration in relation to t= he readout is at error.
Steve Chapman
Senior Consultant E.M. Protrain, Oxford, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
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I have noticed that the tops of my hands are loosing feeling. I was told by another microscopist that it is a common problem with people who run this type of equipment to have symptoms that are not quite, but similar to carpal tunnel symptoms. He said that a few years ago one of the microscopy publications had an article about the problem. Do any of you have any info on this subjesct or are having the same problem? Do you know where I might locate the article. He said that the problem in his case came from ergonomics and was an inflammation of the muscles surrounding the neck. Thanks, Larry Davidson larry.davidson-at-weirton.com
Does anyone have a method to transfer image files between EMS (on a Vax) and Digital Micrograph (on a Mac)? I would be happy to get C code, fortran code, or just inofrmation on the file format of the EMS output files so I can write the code myself (in C). Any code which converts EMS files into raw data would work also. Many thanks Murray Gibson
J. Murray Gibson Professor of Physics and Materials Science Associate Director, Frederick Seitz Materials Research Laboratory **************************************************** *on sabbatical leave until 7/1/98 at * *CNRS-CECM * *15, Rue Georges Urbain * *F94407 Vitry/Seine CEDEX * *FRANCE * *Tel - (0)1.46.87.35.93 (country code 33) * *Fax - (0)1.46.75.04.33 * *Home: telephone Paris - (0)1.40.02.03.51 * *address: 26 Rue Mousset Robert,75012 Paris, FRANCE* **************************************************** e-mail: j-gibson-at-uiuc.edu (unchanged) Permament address: Frederick Seitz Materials Research Laboratory University of Illinois, 104 S. Goodwin Ave (Room 258) Urbana, IL 61801 Tel: (217)-333-2997; Fax: (217)-244-2278; j-gibson-at-uiuc.edu
} ---------- } From: GoYakKlah[SMTP:thetruth-at-southwind.net] } Sent: Tuesday, March 10, 1998 10:02 AM } To: paqui-at-iris1.fae.ub.es; Microscopy-at-Sparc5.Microscopy.Com; } Microscopy.com-at-Sparc5.Microscopy.Com } Subject: size of Electron Microscope } } Im sorry if this is a REALLLLL Dumn question.......... But just what } is the } size of some of these TEM,SEM ect } Ron Mayer } thetruth-at-southwind.net } Simply, they are all "bigger than a breadbox".
} Bruce F. Ingber } Biologist- Electron Microscopy } USDA-ARS, SRRC } 1100 Robert E. Lee Blvd. } New Orleans, LA 70124 } } (504) 286-4270; fax (504) 286-4419 } bingber-at-nola.srrc.usda.gov } } } } }
Try the MAS server at: ftp://www.microanalysis.org/Pub/MMSLib/EMPA/CITZAF3/ Scott
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
According to my doctor, when I experienced this to include "tingleing" in my right arm, this was a posture induced problem of a nerve being slightly pinced in the neck from being hunched over the microscope. I was concious of my posture, scope and chair placement, and perforned some simple (tuck chin in hard) neck exercises and the problem went away. And the problem went away went away.
Good luck and shalom from Jerusalem. Azriel
On 11 Mar 1998 Larry.Davidson-at-weirton.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } I have noticed that the tops of my hands are loosing feeling. I was } told by another microscopist that it is a common problem with } people who run this type of equipment to have symptoms that are not } quite, but similar to carpal tunnel symptoms. He said that a few } years ago one of the microscopy publications had an article about } the problem. Do any of you have any info on this subjesct or are } having the same problem? Do you know where I might locate the } article. He said that the problem in his case came from ergonomics } and was an inflammation of the muscles surrounding the neck. } Thanks, } Larry Davidson } larry.davidson-at-weirton.com }
Yes, I have had pain in the upper arms and numbness in my thumbs due to the C-5 nerve branch getting pinched from the poor ergonomic neck position from years of microscopy. I wear it like a badge of honor. March on! fellow microscopist! I want a microscope that conforms to a person sitting in an old barbers chair.
Bob Underwood Derm Imaging Center U of Wash.
On 11 Mar 1998 Larry.Davidson-at-weirton.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } I have noticed that the tops of my hands are loosing feeling. I was } told by another microscopist that it is a common problem with } people who run this type of equipment to have symptoms that are not } quite, but similar to carpal tunnel symptoms. He said that a few } years ago one of the microscopy publications had an article about } the problem. Do any of you have any info on this subjesct or are } having the same problem? Do you know where I might locate the } article. He said that the problem in his case came from ergonomics } and was an inflammation of the muscles surrounding the neck. } Thanks, } Larry Davidson } larry.davidson-at-weirton.com }
A colleague of mine is looking for a used microtome to section materials (especially paper) for light microscopy for a QC operation. Anybody with such equipment can contact me directly. Thanks in advance, Lynne Garone Polaroid Corp. 781-386-1446 GaroneL-at-Polaroid.com
-----summary------ Does anyone have ideas on VERY low contrast matrix materials in which I could embed particulate materials for such an analysis? In other words, I do not want to see the matrix at all. But it needs to be present to hold the particles in their orientation. (Sometimes you want to see the matrix but in my current case I do not.)
Thanks in advance -
Tyler C. Gruber, Physicist
-----details------- I analyze the structure of nanoscale particles by themselves and also mixed and embedded in polymers and other "matrices". Naturally, the particles are often difficult to measure accurately via any means due to the presence of the matrix. It is the matrix materials, not the subsequent analysis techniques, that I need help with.
I have what I think is a somewhat unusual problem.
Ideally, I would like to image some nanoscale particles in a matrix that is as close as possible to completely transparent to the electron beam. This would enable me to view the particles with nearly the clarity I get when I simply disperse them on a carbon-coated grid.
I want to do this because I am interested in experimenting with changes in the way I process compounds to see if this has influence on the compound isotropy in terms of the orientation of the nanoscale particles (they are non- spherical). It would be proof of concept for other matrices - that anisometry in particle orientation can be detected.
Once embedded I envision microtoming or thinning the compound to achieve a thickness that would render the contribution of the matrix to the image insignificant compared to that of my particles. Therefore, it would be very desirable for the material I am pursuing to be processible in some such manner.
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Ice machines 3/11/98 11:34 = AM
Dear Microscopists: Does anyone know the telephone number for RossTemp in Mason City, Iowa, = USA? Or maybe someone has information as to where I could purchase an = industrial ice machine/maker. Any help would be appreciated greatly. = Thank you. Linda Chicoine Center for Cell Imaging Dept. of Cell Biology Yale University New Haven, CT USA 203-785-3646
with ESMTP id {01IUJB66T6MY95MPWC-at-asu.edu} for microscopy-at-Sparc5.Microscopy.Com; Wed, 11 Mar 1998 09:58:49 MST Received: from csss.la.asu.edu (csss2.la.asu.edu [129.219.54.220]) by smtp1.asu.edu (8.8.7/8.8.7) with ESMTP id JAA21509 for {microscopy-at-Sparc5.Microscopy.Com} ; Wed, 11 Mar 1998 09:59:05 -0700 (MST) Received: from default (129.219.100.199) by csss.la.asu.edu with SMTP (Apple Internet Mail Server 1.1); Wed, 11 Mar 1998 09:54:49 -0700
Dear All: I need to prepare a cross-section specimen out of a structure formed on a bulk single-crystalline diamond substrate (approx 300um thick). Does anyone has a relevant experience? Any advice will be appreciated. I tried to cut it with a slow-speed diamond disk-saw without much success... TIA,
Max Sidorov
------------------------------- Maxim V. Sidorov, Ph.D. Center for Solid State Science Arizona State University Tempe, AZ 85287-1704 USA Phone: (602)727-6260 Fax: (602)965-9004
I'm the electron microscopist at Eastern Connecticut State University. A new science building is going to be built here and I've been given the task of making recommendations for the new electron microscope laboratory. I would be interested in communicating (and perhaps visiting) with anyone who has recently been involved in designing a microscopy lab from the ground up. I was told to design for the future; to build a lab that will be functional 20 years from now. I want to get information not only on the best design for the physical structure, but also advice on equipment.
Also, if there are any vendors out there who might be interested in having a room, hallway, wing, or even the entire building named in their honor (these are our president's words), we would love to discuss with you how your gifts to the university could make that happen.
Marty Levin
Martin A. Levin Department of Biology Eastern Connecticut State University Willimantic, CT 06226 Phone: (860)465-4324 Fax: (860)465-5213
Dear Daniel, } } My daughter prepares a thesis on artistic paintings and would like to know } if there are recent publications on the use of the SEM and EDX on } restoration of painting, expertise or research in history of art. } Mc Crone has done this sort of work (and there may be many others). In a closely related field, Tom Cahill has used the proton beam from the cyclotron to do EDX on many specimens including a Gutenberg Bible. This technique is almost the same as EDX on a scope or probe, but the atoms are excited or ionized by protons, so the brehmsstrahlung background is much lower. Proton optics are not as highly developed as electron optics, so the spatial resolution of PIXE (particle-induced x-ray emission) with protons is not as good as that with electrons. Yours, Bill Tivol
We have recently been faced with an issue of analyzing cross-sections of 1mm thick polycrystalline diamond disks for ASEM and electron microprobe analyses. The initial sectioning was done with EDM. The cross-sections were then mounted in bakelite and metallographically polished.
For ATEM, you might be able to use the EDM technology to section a piece sufficiently small for an ion dimpler to take down to the desired thickness. We are currently addressing potential preps for a possible ATEM analysis of this material.............I'll let you know what shakes out of our discussions........
Cheers and Good Luck
Cary Black Dow Chemical phone: (517) 636-5760 e-mail: ckblack-at-dow.com
Dear Tyler, } } Does anyone have ideas on VERY low contrast matrix materials in which } I could embed particulate materials for such an analysis? In other words, I } do not want to see the matrix at all. But it needs to be present to hold the } particles in their orientation. (Sometimes you want to see the matrix but in } my current case I do not.) } The contrast in EM comes from the elastically scattered electrons, so the lower the scattering cross-section, the greater the transparency, and the more uniform the matrix, the lower the contrast. You should have the best results with a matrix having the fewest electrons per unit volume.
} I analyze the structure of nanoscale particles by themselves and also } mixed and embedded in polymers and other "matrices".
These polymers would seem to be ideal. If the particles are of nearly the same composition, however, you will have to stain them with a heavy metal which does not also stain the matrix polymer. If the par- ticles are themselves heavy metals or other higher-Z-than-carbon materials they should be visible without additional processing. Good luck. Yours, Bill Tivol
We are still consolidating and would like to see this good, but old equipment put to use somewhere. You may have all this equipment for free if you are a DOD agency, Federal Government, a State Government agency, or a State Educational Institute. Contact me, Thomas A Baginski via Email or phone 301 295 5691 or our logistics officer Mr Jerry Sherman -at- 301 295 3435. We have plans to clear out everythihng by March 20th, 1998. JEOL Scanning 35U with BEI, TOPOG, ETC, service reports, much more Hummer Carbon Evaporator unit, and Sputter Coater Haskris Chiller, for above Balzer's Spray Freezing Unit, 15 years old, never used Ultrastable Microtomy table TOUSIMIS Critical Point Dryer MANY, other assorted peripheral equipment that can be used with al the above. Our GSA department is ready to do the paperwork, NOW.... Good luck. Tom Baginski
Greetings to All ... I am currently using a Kodak Ektamatic Photo Processor that is no longer manufactured (as far as I know). It's sort of on its last leg and I HATE to see it go. I would love to find another one just like it (you know, so I won't have to change the status quo!). Is there anyone out there that might have one in storage that you might be willing to part with? If not, how about pointing me in the right direction to one that uses the same S-II Activator and S-30 Stabilizer chemicals. I get such nice crisp glossy photos now and don't want that to change.
Thanks in advance for any information ...
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Fort Worth Fort Worth, Texas
SPIRATONE makes a considerably cheaper yet comparable stabilization processor along with chemicals and paper. I believe you might be able to use the Ektamatic chemicals in it - but not completely sure about this. Do a WWW search for Spiratone Stabilization Processor. I used one for many years and was well pleased with it.
} I am currently using a Kodak Ektamatic Photo Processor that is } no longer manufactured (as far as I know). It's sort of on its last leg } and I HATE to see it go. I would love to find another one just like it } (you know, so I won't have to change the status quo!). Is there anyone } out there that might have one in storage that you might be willing to } part with? If not, how about pointing me in the right direction to one } that uses the same S-II Activator and S-30 Stabilizer chemicals. I get } such nice crisp glossy photos now and don't want that to change.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I think that you have two immediate options and perhaps a far out option.
1. FIB comes to mind right away, but I don't know how well the diamond will mill. Argon does not do well with carbon, so I don't expect Ga to do any better. You could do it right from your thick sample.
2. 300 um is a little thick, but you might try cleaving the sample. Look at our paper on the small angle cleavage technique in Vol 480 of the MRS Symposium Proceedings (1997) to get an idea on how it is done for semiconductors and see if it might be modified for your thicker and harder samples. I think that you would have to know the crystallographic directions pretty well before you start. You might consider contacting a jeweler to find a diamond cutter to help you cleave it and/or polish it down thinner. If you get the sample down to 80-90 um, I think you have a chance with SACT because it works for SiC single crystals.
Far out option: Peter Humble once told me that they drilled holes in diamond foils in an old TEM because the vacuum conditions were poor. I think that the reason was that it was due to water vapor, carbon, and the electron beam reacting at the diamond surface. Well, how about if you find an environmental SEM, backfill with H20 vapor, crank up the voltage and current density to the highest value and see if you can mill the sample out in the same way as the FIB does. Contact Philips. They have the FEG ESEM and see if they are game to try it. Let me know if it works.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Dear All: I need to prepare a cross-section specimen out of a structure formed on a bulk single-crystalline diamond substrate (approx 300um thick). Does anyone has a relevant experience? Any advice will be appreciated. I tried to cut it with a slow-speed diamond disk-saw without much success... TIA,
Max Sidorov
------------------------------- Maxim V. Sidorov, Ph.D. Center for Solid State Science Arizona State University Tempe, AZ 85287-1704 USA Phone: (602)727-6260 Fax: (602)965-9004
I inherited a Joel 733 that we need to clear out for new equipment. The microscope portion of the instrument including the automated stage was working but the data system was not. Because of the data system not working, the condition of the 4 WDS detectors and one Germanium EDS detector is unknown. According to the documentation the Noran system software is "Series II Software Release 1H" the disks are labeled TN-550-220 Volume #20, #21, #22, And #26.
No reasonable offer will be refused. If you are interested please e-mail me privately.
Mike
=========================================================== Michael Dunlap lab (530) 752-0284 Facility For Advanced Instrumentation fax (530) 752-4412 University of California mrdunlap-at-ucdavis.edu Davis CA, 95616 http://carbon.ucdavis.edu ============================================================
We prepared some small (around 1 mm) diamond samples a few years ago and examined in the TEM. We mounted them in Buehler's Epomet mounting compound, and were then able to cut them successfully on a low-speed diamond saw. Once cut, they were dimpled. We have a South Bay dimpler, and they sold us a special diamond-impregnated dimple-grinding wheel for hard materials that we just had to lubricate. This method worked fine for our material. [No financial interest etc.]
Gill Bond Dept Materials & Met. Eng. New Mexico Tech
On Wed, 11 Mar 1998, Max Sidorov wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All: } I need to prepare a cross-section specimen out of a structure formed on a } bulk single-crystalline diamond substrate (approx 300um thick). Does anyone } has a relevant experience? Any advice will be appreciated. } I tried to cut it with a slow-speed diamond disk-saw without much success... } TIA, } } Max Sidorov } } ------------------------------- } Maxim V. Sidorov, Ph.D. } Center for Solid State Science } Arizona State University } Tempe, AZ 85287-1704 USA } Phone: (602)727-6260 } Fax: (602)965-9004 }
We use 1% agar to hold tissue culture cells together. In thin sections, it's barely visible. Would this work for you?
On Wed, 11 Mar 1998, TylrGruber wrote:
} Date: Wed, 11 Mar 1998 11:19:41 EST } From: TylrGruber {TylrGruber-at-aol.com} } To: Microscopy-at-sparc5.microscopy.com } Subject: No Subject } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } -----summary------ } Does anyone have ideas on VERY low contrast matrix materials in which } I could embed particulate materials for such an analysis? In other words, I } do not want to see the matrix at all. But it needs to be present to hold the } particles in their orientation. (Sometimes you want to see the matrix but in } my current case I do not.) } } Thanks in advance - } } Tyler C. Gruber, Physicist } } -----details------- } I analyze the structure of nanoscale particles by themselves and also } mixed and embedded in polymers and other "matrices". Naturally, the } particles are often difficult to measure accurately via any means due to the } presence of the matrix. It is the matrix materials, not the subsequent } analysis techniques, that I need help with. } } I have what I think is a somewhat unusual problem. } } Ideally, I would like to image some nanoscale particles in a matrix that is as } close as possible to completely transparent to the electron beam. This would } enable me to view the particles with nearly the clarity I get when I simply } disperse them on a carbon-coated grid. } } I want to do this because I am interested in experimenting with changes in the } way I process compounds to see if this has influence on the compound isotropy } in terms of the orientation of the nanoscale particles (they are non- } spherical). It would be proof of concept for other matrices - that anisometry } in particle orientation can be detected. } } Once embedded I envision microtoming or thinning the compound to achieve a } thickness that would render the contribution of the matrix to the image } insignificant compared to that of my particles. Therefore, it would be very } desirable for the material I am pursuing to be processible in some such } manner. } } Thanks in advance - } } Tyler C. Gruber, Physicist } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America Kiymetli Julide Hanim, C.ALLAH kavustursun... Buyuklerinizin Hac Ibadetini C.ALLAH kolaylastirsin , Insaallah... Asagida gonderdigim mesaji Zulal Hanima veya Malzeme'de Elektron Mikroskoplarla ugrasan baska kim varsa , verebilirmisin, Lutfen... C.ALLAH'tan sana, ailene ve tum sevdiklerine saglik, huzur ve basarili gunler niyaz ediyoruz... Selamlarin en guzeliyle... Yuzer ailesi.
Sayin Zulal Hanim,
ListServer-at-MSA.Microscopy.Com adresine yazildiginizda Size bedava Mikroskopi hakkinda bu konoda uzman olan ve bu adresle yazistiklari kisilerin mesajlari gelecektir... Ornek bir mesaj asagidadir... Selamlarin en guzeliyle... Hayrettin Yuzer.
Dear Tyler, } } Does anyone have ideas on VERY low contrast matrix materials in which } I could embed particulate materials for such an analysis? In other words, I } do not want to see the matrix at all. But it needs to be present to hold the } particles in their orientation. (Sometimes you want to see the matrix but in } my current case I do not.) } The contrast in EM comes from the elastically scattered electrons, so the lower the scattering cross-section, the greater the transparency, and the more uniform the matrix, the lower the contrast. You should have the best results with a matrix having the fewest electrons per unit volume.
} I analyze the structure of nanoscale particles by themselves and also } mixed and embedded in polymers and other "matrices".
These polymers would seem to be ideal. If the particles are of nearly the same composition, however, you will have to stain them with a heavy metal which does not also stain the matrix polymer. If the par- ticles are themselves heavy metals or other higher-Z-than-carbon materials they should be visible without additional processing. Good luck. Yours, Bill Tivol
................................................................................ Dr. Hayrettin Yuzer Visiting Researcher Research Center for Interface Quantum Electronics Hokkaido University Nort 13 West 8, Sapporo 060, JAPAN Tel: +81-11-706-7176 (w) +81-11-707-3715 (h) Fax: +81-11-716-6004 E-mail: yuzerh-at-ryouko.rciqe.hokudai.ac.jp
I came across this great application and thought you might be interested... it allows you to easily record virtually anything on your PC screen: applications, browsers, desktop activities, etc. and turn it into a self-running digital movie! It's great for web or CD tutorials, demos, engineering communications, customer service and a lot more. It definitely gets the point across quickly! You can see examples and more at www.infosynth.com
Hope to talk to you soon!
Jennifer
P.S. If you like, I'll send you more details and a copy of the full-function demo. Just drop me a line, or you can download it from the website! Bye!
jenn-at-infosynth.com
_________________________________________________ Information Synthesis, Inc. 8105 S.W. Boeckman Road Wilsonville, Oregon 97070 503/685-0302 800/547-3016
OK , I know everyone doesn't have one but just in case you do & nothing else works...... I recall a few years ago at Rice University that Bob Houge had students ablating holes in diamond chips with a KrF excimer laser (248nm). They were monitoring progress with a interferometer so I'd say the level of control was pretty good (although it was not real quick). No idea what their objective was but Bob is friendly. I don't have an e-mail address but link into www.rice.edu } Chem. dept.} Prof. John Margrave & see if you can zero in on Bob. If that doesn't work , drop me a line & I'll look him up while I am on campus.
Your best bet may be to either reduce your accelerating voltage or look into a good backscatter detector. If you have the money, you also might want to look into an ESEM, the partial gas atmospheres may be enough to drain off the charging you experience.
} Previous posts are true for larger particles which are good } insulators. On the other hand, I often need to examine mounted } and polished finely divided conductive and semi-conductive } material. Quite often I am looking for hi-Z metal carbides. } Carbon coating really hurts the desired to } undesired/signal/noise ratio. } } It is also true that conductor loaded insulating resins (bulk } conducting) do not help much in this area. The islands of } charging resin will "eat you alive" {g} . } } To clarify my earlier post: } } What I would like to see developed is intrinsically conductive } polymer (sometimes know as unobtianium) that can be used to } mount and polish specimen material (fines). This sort of } polymer is currently available in thermoset (already) powder } and, as another poster mentioned, solid forms (block, sheet, } etc.). This polymer itself is conductive. Several different } schemes may be used. One method, for example, adds an iodine } atom in the polymer chain to free some conductance electrons. } } Woody White } McDermott Technology, Inc. } } } I agree with Jean Marc. All this excitement about conducting resins } is not likely to achieve the aim, which was the elimination of } charging in the SEM of DIFFICULT non-conductors embedded in resins. } {snip} } } Jim Darley } } ProSciTech Microscopy PLUS PO } Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 } Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, } MSDS, User Notes **************************** } www.proscitech.com.au ***** } -------------------------------------------------------------------- } ---. } } } Just a remark on conductive resins: they are good to embed } conductive samples for surface SEM observation for example. But } don't forget, they are composite materials, i.e. non-conductive } resin loaded with enough conducting material (Cu, Ag whatever) to } make them MACROSCOPICALLY conductive. } } } In microscopy application, their composite structure quickly appears } and limit their usage. For example, their are useless to look at } edges of conductive material because the non-conductive resin } portion in-between the conductive particles will charge up exactly } as pure polymer and blur out the picture. } } For your ion-milling application I suspect the same holds true but I } haven't } } try myself. So just be aware of this } problem if doesn't work. } } BTW they are available from any electron microscopy accessories } vendor. } } } Jean-Marc Boichat email: jean-marc.boechat-at-chma.mhs.ciba.com } EM LABS FO 5.1 phone:+4126 435 6979 fax: +4126 435 6907 } Ciba research Center } P.O. Box 64 } CH- 1723 Marly 1 When things go wrong, don't go with them! } Switzerland } } Disclaimer: "nobody in this company ever cared for what I said, why } would they start now". } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Maintening the microscope itself I have problem with vacuum tightness of the (for me - so called ,,third party'' option) Oxfords-LINK Pentafet MK6b type detector unit. Thats the one attached to TEM and is retractable with UTW which can not outstand the vented column so has the pneumatic shut-of valve and motorised retraction mechanism. Like LINK has in tradition there is 3-section steel bellow. The problem is that after opening the valve the vacuum goes wrong - somwhere leak in the detector. As there is limited space around, and service flange to connect quadrupol is quite far - I don't see real possibility to trace such leak precisly with helium leak detector. The leak is on the IGP vacuum level - so very small - but enough to shut down the microscope.
1) Does anybody know f.e. procedure how to admit air to the detector to equalize pressure on the both sides of the window and dismantle and change the Oring in the shut-off valve ? that one is most suspected. 2) How long can this type of detector be not cooled waiting for solution ? 3) is it better to keep it not-cooled when there is a leak waiting for service, or to keep it cooled - what in my opinion can cause intensive cryo-pumping effect and contaminate parts inside, and after warming up condensate vapours etc. ? 4) Are there any firms who would take such detector for repair/recheck/retight servicing ??? I am asking above question knowing the manufacturers price for such repair (generally leading to the ,,better buy new one" conclusion) and expecting better conditions.
kind regards
Krzysztof Herman FEI/PHILIPS Service Labsoft, Domaniewska 9/11/45, 02-663 Warszawa tel: (48 601) 307456, fx:(48 22) 483787 E-mail: kherman-at-labsoft.com.pl http://www.labsoft.com.pl/
Email: knechtew-at-tcd.ie Name: Walter Knechtel School: Trinity College Dublin, Ireland
We are using formvar as a substrate for TEM investigations. We are studying charging effects on it. Is there any possibility to estimate quantitativly how much the substrate charges up when exposed to the electron beam? The current density of the beam is known.
} ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Max: As difficult as it may seem, diamond can be microtomed with a diamond knife to prepare cross-sections for TEM analysis (as long as you don't require a cross-section of the full 300 um). I've had a great deal of experience and luck cross-sectioning hard materials using the technique. You can review an ultramicrotomy procedure for cross-sectioning diamond and cubic boron nitride in "Ultramicrotomy of Diamond Films for TEM Cross-section Analysis," P. Swab, Microscopy Research and Technique, Wiley-Liss Inc., vol. 31, pp. 308-310 (1995). The paper describes the formation of "micro chips" that are preferentially oriented, embedded in epoxy, and then cross-sectioned with a diamond knife. These cross-sections may show mechanical artifacts, but are uniformly thick and free of beam damage and chemical artifacts.
} Regards, } } Phil Swab } Advanced Coatings Division } Advanced Refractory Technologies } Buffalo, NY, USA } Phone: 716-875-4091 } } } On Wed, 11 Mar 1998, Max Sidorov wrote: } } } } ---------------------------------------------------------------------- } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } ---------------------------------------------------------------------- } -. } } } } Dear All: } } I need to prepare a cross-section specimen out of a structure formed } on a } } bulk single-crystalline diamond substrate (approx 300um thick). Does } anyone } } has a relevant experience? Any advice will be appreciated. } } I tried to cut it with a slow-speed diamond disk-saw without much } success... } } TIA, } } } } Max Sidorov } } } } ------------------------------- } } Maxim V. Sidorov, Ph.D. } } Center for Solid State Science } } Arizona State University } } Tempe, AZ 85287-1704 USA } } Phone: (602)727-6260 } } Fax: (602)965-9004 } } }
I notice that Olympus are advertising 'Pain-free microscopy' on their BX40 microscope. It is supposed to incorporate new ergonomic features such as an improved shape, focussing controls near to bench height and adjustable position of eyepieces. I have had no experience of this machine and have no connection with the company, other than as a satisfied user of some of their teaching and research microscopes. It is to be hoped that most of the leading manufacturers are doing something like this.
Malcolm Haswell Electron Microscope Unit University of Sunderland UK ----------
hello all-
i've got a problem with the vacuum system on a JEOL35. the problem seems to manifest itself by disabling the automatic valve sequencing function. i can operate all the valves in manual mode but in auto mode it never seems to reach vacuum ready. in manual mode it pumps OK, but manually operating the airlock valves is a pain.
any ideas why manual sequencing the valves works and automatic doesn't??
thx brian
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
We have a researcher who needs a silicon oil which turns to a jelly in UV light - for sticking down crystals with good thermal contact at cryogenic temperatures. Ordinary silicon oil, as for diffusion pumps, won't work because it cracks at low temperatures and isn't strong enough. Any ideas? Thanks Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
Greetings to all. To help get the word out, I've been asked by our Institute for Agriculture and Natural Resources to post the following announcement of a new position in a new biomicroscopy facility at the University of Nebraska-Lincoln. Best wishes, Brian
------------------------------ MICROSCOPY FACILITY MANAGER: RESEARCH ASSISTANT PROFESSOR
Immediate opening for non-tenure track Research Assistant Professor with experience in Fluorescence, Scanning Confocal and Light Microscopy. Individual will co-manage new Center for Biotechnology Microscopy Core Research Facility serving research, teaching and industrial clients located in the new George W. Beadle Center for Genetics and Biomaterials Research. Technical support provided. Doctorate required. Knowledge of computer-based image analysis, data storage, transmittal and networking essential. Extremely important that successful candidate is able to work effectively with people with diverse research interests and provide expertise and service. Individual will be expected to develop strong research collaborations with UNL scientists. Research professorial position will be administered through a specific academic unit. Screening of applications will begin May 1, 1998, and continue until suitable candidate is identified. Apply to: Dr. James Kinder, Center for Biotechnology, University of Nebraska, N300 Beadle Center, Lincoln, NE 68588-0665. Phone: 402/472-6438. E-Mail: ansc502-at-unlvm.unl.edu. Web: http://www.biotech.unl.edu. UNL is committed to a pluralistic campus community through Affirmative Action and Equal Opportunity; is responsive to the needs of dual career couples; and assures reasonable accommodation under the Americans With Disabilities Act. Contact Dr. Kinder for additional information.
-------------------------------------
*********************************************************** Brian W. Robertson *Office 402 472 8308 Assoc. Prof. *FAX 402 472 1465 Department of Mechanical Engineering *Lab 402 472 8762 and Center for Materials Research and Analysis University of Nebraska-Lincoln N124 WSEC, Lincoln, NE 68588-0656, USA ***********************************************************
JXA50A from JEOL has variable resistors on its vacuum board for setting thresholds for automatic mode. I am sure, the JSM35 has the same (they share many boards); see manuals for vacuum board. Good luck!
Vladimir Dusevich NCSU
On Thu, 12 Mar 1998, Brian McIntyre wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } hello all- } } i've got a problem with the vacuum system on a JEOL35. the problem seems } to manifest itself by disabling the automatic valve sequencing function. i } can operate all the valves in manual mode but in auto mode it never seems } to reach vacuum ready. in manual mode it pumps OK, but manually operating } the airlock valves is a pain. } } any ideas why manual sequencing the valves works and automatic doesn't?? } } thx } brian } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627 } } 716-275-3058 } 716-244-4936(fax) } } }
Hello all I need some advice on replication. We want to determine the depth of a conductor a few um below the surface. Interferometry has give me the depth relative to the immediate edge but we need to know the distance from the highest point on the surface. The surface is nominally 1cm Sq. with a slit .6mm X ~3um across it. I was thinking of something that once formed could be cut, polished & measured. Ideas?
I am looking for the best way to upgrade the old Kevex uX 7000 EDS system on my Amray 1000A SEM. (Unfortunately a big part of "best" is a low price.) I've been looking at the 4pi Spectral Engine II package for EDS, digital image acquisition, and x-ray mapping. The users I have talked to have all sounded quite happy with their systems and the price is good. I know there are an awful lot of other companies out there offering EDS upgrade packages though and we need to be satisfied that we're making the right choice.
Please let me know what your experiences are with EDS upgrades. I'd be happy to hear from vendors as well. If you will respond directly to me, rather than to the list, I'll post a summary of the responses I get.
I mostly analyze small particles (0.1 to 1 micron) in tissue sections or on filters so we don't need extravagant quantitative capabilities. I plan to use my existing detector, preamp, pulse processor, and bias supply. I have no strong preference between Mac and PC.
Thanks in advance for your help, Tori Hatch
thatch-at-hsph.harvard.edu 665 Huntington Avenue, II-227 Boston, MA 02115 (617) 432-4429
Does anyone have first-hand experience in working with Nanoplast (Melamine resin) in conjuntion with immuno-gold staining? What, if any, are the unseen pitfalls in its use?
Thanks, Steve Schmitt
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
This is a general reply to the bunch of you who provided me with info on how to contact CM Taylor.
Basically, it goes like this:
1. the address is:
C.M. Taylor Company 71 Bonaventura Drive San Jose, CA 95134 U.S.A. ph: 408-526-9020 fx: 408-526-9021
2. the best contact is the General Manager, Jerry V. Weatherford, who is more than willing to answer questions!
3. the company has a brochure describing the compounds and metals available through them, as well as mounting formats they can provide.....their prepared standard mounts are called Taylor Mutli-Element Standards (TM-ES), and range in price from $500 to $4200, depending on content and format
4. in regard to the metals: most metals are 99.99% pure, or greater (99.999%) - the only exception is the Hf metal which contains 1.65% Zr (noted as 3.32% in older literature; could be, however, a different standard, now replaced).
Hope this helps.
Winton
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
I had saved an e-mail from about 2 years ago, in which is given a number for a company in CA (MicroDataWare) that one could contact to get the McCrone Particle Atlas on CD-ROM. Today - yep, two years later! - I called the number, and, no surprise - no longer at that number!
My question toall of you has two parts:
1. does anyone out there know where I can get a copy of the Atlas on CD-ROM? (I have heard the paper copy is long out of print), and
2. have any of you used the Atlas?....if so, how about comments on its practical use (as opposed to the "it's nice to look at" side of it).
Thanks, in advance, for any feedback.
Winton Cornell
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
} Folks: } } I had saved an e-mail from about 2 years ago, in which is given a number } for a company in CA (MicroDataWare) that one could contact to get the } McCrone Particle Atlas on CD-ROM. Today - yep, two years later! - I called } the number, and, no surprise - no longer at that number! } } My question toall of you has two parts: } } 1. does anyone out there know where I can get a copy of the Atlas on } CD-ROM? (I have heard the paper copy is long out of print), and } } 2. have any of you used the Atlas?....if so, how about comments on its } practical use (as opposed to the "it's nice to look at" side of it).
Winton, I can answer the first question. I'll leave it to others to answer the second. MicroDataware has beeen in limited operation over the past two years as I've recovered from some serious health issues. (Better now!) The Particle Atlas Electronic Edition (PAEE) is available from two sources in the USA:
McCrone Research Institute, Chicago, IL 1-312-842-7100 attention: Nancy Daerr
McCrone Accessories and Components, Westmont, IL 1-630-887-7100 attention: (anyone at MAC)
The print Atlas has been out of print for many years now. Thanks for your interest in the Atlas and I hope you find it useful. If you need any further information please let me know off list.
Steve Shaffer MicroDataware sshaffer-at-microdataware.com
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction in limited service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
Regarding the use of Nanoplast in conjunction with immuno-gold, I have some experience with this. The resin is very interesting from an ultrastructural perspective. Given its solubility in water, we found the presence of many connective tissue components after embedding in Nanoplast which otherwise would have been lost after dehydration (proteoglycans in particular). We were very excited to use it for section-surface immunocytochemistry. Our first attempts were very promising. However, we occasionally got a very specific labeling pattern to structures which we were absolutely sure did not contain the antigen! Puzzled, bothered, and frightened, we put it aside for a while.
Best of luck,
Doug
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
On Wed, 11 Mar 1998 16:53:03 -0600 "John J. Bozzola" {bozzola-at-siu.edu} wrote:
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- Send Email } to } ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anyone have first-hand experience in working with } Nanoplast (Melamine resin) in conjuntion with immuno-gold } staining? What, if any, are the unseen pitfalls in its use? } } Thanks, } Steve Schmitt } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } #################################################################### } }
JXA50A from JEOL has variable resistors on its vacuum board for setting thresholds for automatic mode. I am sure, the JSM35 has the same (they share many boards); see manuals for vacuum board. Good luck!
Vladimir Dusevich NCSU
} On Thu, 12 Mar 1998, Brian McIntyre wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } hello all- } } } } i've got a problem with the vacuum system on a JEOL35. the problem seems } } to manifest itself by disabling the automatic valve sequencing function. i } } can operate all the valves in manual mode but in auto mode it never seems } } to reach vacuum ready. in manual mode it pumps OK, but manually operating } } the airlock valves is a pain. } } } } any ideas why manual sequencing the valves works and automatic doesn't?? } } } } thx } } brian } } } } **************************************************************** } } Brian McIntyre } } Electron Microscopy Lab } } Institute of Optics } } University of Rochester } } Rochester, NY 14627 } } } } 716-275-3058 } } 716-244-4936(fax) } } } } } } } }
Position available, applications invited from US and overseas applicants.= =0APlease send resume as an attachment to be considered or e-mail for fur= ther=0Ainformation to Mark1306-at-aol.com
Position: Reliability Test Manager (Southern California)
Description:: This individual will be responsible for: =95Managing the Reliability Assurance Testing and Failure Analysis Labs.= =A0 =95Provide electrical, physical and environmental performance data on IC semiconductor devices. =95Performing failure analysis and evals on performance failures from newly designed products, on-going reliability assessment programs and customer field failure returns. =95Determining failure modes root causes and issuing formal technical reports to inside support staff. =95Provide sales, marketing and customer support for qualification, reliability, failure analysis and failure rate data as needed.=A0
Min 5 yrs. hands-on experience with: =95Failure analysis and evaluation techniques of IC devices including the=
use of optical microscopy, scanning electron microscopy (SEM), energy dispersive x-ray analysis (EDX), radiographic inspection, decapsulation, sectioning and polishing.=A0 =95Reliability/qualification testing of IC=92s include operational knowle= dge of various manual & auto-programmable electrical, physical and environmental test equipment IAW Military and Industry Test Standards, i.e. MIL-STD-750, 883 EIA, JEDEC & ASTM publications. =0A
I think in your case may be caused by the offset voltage of vacuum control circuit is shifted. You need to re-adjust a high vacuum control signal (ready lamp light on). Check at vacuum volage measuring terminal by multimeter after main valve open it should decreasing from about 160uA to 0 uA within few minutes and then the ready lamp will come if not adjust vacuum comparator circiut in vaccum control.
Regards,
Paiboon Nuannin
Prince of Songkla University Hatyai, Thailand
On Thu, 12 Mar 1998, Brian McIntyre wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } hello all- } } i've got a problem with the vacuum system on a JEOL35. the problem seems } to manifest itself by disabling the automatic valve sequencing function. i } can operate all the valves in manual mode but in auto mode it never seems } to reach vacuum ready. in manual mode it pumps OK, but manually operating } the airlock valves is a pain. } } any ideas why manual sequencing the valves works and automatic doesn't?? } } thx } brian } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627 } } 716-275-3058 } 716-244-4936(fax) } } }
I think in your case may be caused by the offset voltage of vacuum control circuit is shifted. You need to re-adjust a high vacuum control signal (ready lamp light on). Check at vacuum volage measuring terminal by multimeter after main valve open it should decreasing from about 160uA to 0 uA within few minutes and then the ready lamp will come if not adjust vacuum comparator circuitt in vaccum control.
Regards,
Paiboon Nuannin
Prince of Songkla University Hatyai, Thailand
On Thu, 12 Mar 1998, Brian McIntyre wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } hello all- } } i've got a problem with the vacuum system on a JEOL35. the problem seems } to manifest itself by disabling the automatic valve sequencing function. i } can operate all the valves in manual mode but in auto mode it never seems } to reach vacuum ready. in manual mode it pumps OK, but manually operating } the airlock valves is a pain. } } any ideas why manual sequencing the valves works and automatic doesn't?? } } thx } brian } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627 } } 716-275-3058 } 716-244-4936(fax) } } }
I tried to handle with Nanoplast (including immunolocalization with colloidal gold) some years ago. I recognized that resin as very interesting having some diferrent characteristics in comparison with other "classical" resins. Please, look into publication:
Weyda F., 1990: Notes on the use of Nanoplast FB 101 for transmission electron microscopy. J.Electron Microsc.Tech., 16: 356-357
Good luck!
Sincerely, Frantisek Weyda
------------------------------------------------------------------------ RNDr.Frantisek Weyda,CSc. | Frantisek Weyda, PhD. Entomologicky ustav AV CR | Institute of Entomology Branisovska 31 370 05 Ceske Budejovice Ceska republika | Czech Republic --------------------------------------------------------------------- tel.: (38) 777 linka 5257 | phone: +420 38 777 ext.5257 fax.: (38) 43625 | Fax.: +420 38 43625
Internet e-mail: weyda-at-entu.cas.cz www sites: www.entu.cas.cz ural.bf.jcu.cz/tix/dost/bfju/WEYDA.HTM -------------------------------------------------------------------------
Good points. I have the BSE and use as low kV as is practical. Helps sometimes, but not always useful depending on requirements like higher mag/resolution imaging and some x-ray analysis. Woody White
Your best bet may be to either reduce your accelerating voltage or look into a good backscatter detector. If you have the money, you also might want to look into an ESEM, the partial gas atmospheres may be enough to drain off the charging you experience.
} Previous posts are true for larger particles which are good } insulators. On the other hand, I often need to examine mounted } and polished finely divided conductive and semi-conductive
} {snip}
} } Just a remark on conductive resins: they are good to embed } conductive samples for surface SEM observation for example. But } don't forget, they are composite materials, i.e. non-conductive } resin loaded with enough conducting material (Cu, Ag whatever) to } make them MACROSCOPICALLY conductive. } } } In microscopy application, their composite structure quickly appears } and limit their usage. For example, their are useless to look at } edges of conductive material because the non-conductive resin } portion in-between the conductive particles will charge up exactly } as pure polymer and blur out the picture. } } For your ion-milling application I suspect the same holds true but I } haven't } } try myself. So just be aware of this } problem if doesn't work. } } BTW they are available from any electron microscopy accessories } vendor. } } } Jean-Marc Boichat email: jean-marc.boechat-at-chma.mhs.ciba.com } EM LABS FO 5.1 phone:+4126 435 6979 fax: +4126 435 6907 } Ciba research Center } P.O. Box 64 } CH- 1723 Marly 1 When things go wrong, don't go with them! } Switzerland } } Disclaimer: "nobody in this company ever cared for what I said, why } would they start now". } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Position available, applications invited from US and overseas applicants.Please send resume as an attachment to be considered or e-mail for furtherinformation to Mark1306-at-aol.com
Position: Reliability Test Manager (Southern California)
Description:: This individual will be responsible for: =95Managing the Reliability Assurance Testing and Failure Analysis Labs.=20 =95Provide electrical, physical and environmental performance data on IC semiconductor devices. =95Performing failure analysis and evals on performance failures from newly designed products, on-going reliability assessment programs and customer field failure returns. =95Determining failure modes root causes and issuing formal technical reports to inside support staff. =95Provide sales, marketing and customer support for qualification, reliability, failure analysis and failure rate data as needed.=20
Min 5 yrs. hands-on experience with: =95Failure analysis and evaluation techniques of IC devices including the use of optical microscopy, scanning electron microscopy (SEM), energy dispersive x-ray analysis (EDX), radiographic inspection, decapsulation, sectioning and polishing.=20 =95Reliability/qualification testing of IC=92s include operational knowledge of various manual & auto-programmable electrical, physical and environmental test equipment IAW Military and Industry Test Standards, i.e. MIL-STD-750, 883 EIA, JEDEC & ASTM publications.
I do not understand if silicone oil and UV light are a requirement. There is an Apiezon grease which has good thermal conductivity and is in fact used for sticking down sensors etc at cryo temperatures:
CRYO-APPLICATION/VACUUM GREASE Apiezon N is a speciality grease for various cryogenic uses, including for vacuum sealing at very low to warm temperatures were its vapour pressure is very low, making it suitable for high vacuum applications. Type N does not craze or crack at low temperatures, has good thermal conductivity at low temperatures and is ideal for mounting sensors and other devices. It has very low levels of magnetic susceptibility which makes it suitable for some superconductor applications. Cycling between absolute zero and +40C is well tolerated by Apiezon N. It is frequently used as a jointing medium on cryostat flanges where its larger contact area, compared with that of indium wire, provides much better thermal transfer. Applications include cryo-systems on electron microscopes, cryostats, magnetic image resonance systems, sealing of vacuum parts including of glass joints and as a heat sink.
Hope this helps. I am sorry that the disclaimer would sound more like an ad, but this is a very minor item to us. Yes we do stock this grease, I expect that at least a couple of other EM suppliers also do. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
-----Original Message----- } We have a researcher who needs a silicon oil which turns to a jelly in UV } light - for sticking down crystals with good thermal contact at cryogenic } temperatures. Ordinary silicon oil, as for diffusion pumps, won't work } because it cracks at low temperatures and isn't strong enough. } Any ideas? Thanks } Elaine } } } } Dr. Elaine Humphrey } Biosciences Electron Microscopy Facility } University of British Columbia } 6270 University Blvd } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-unixg.ubc.ca } } }
We have a copy of the CD version of the Particle Atlas and use it regularly when identifying contaminants. The search engine in it is reliable and the image quality is pretty good too.
Joe Neilly Microscopy and Microanalysis Abbott Laboratories Abbott Park, IL 60064
Dear Colleagues: I was asked to make a positive print from a slide ( for presentation )with color histology image ( positive image). I have a ScanMaker III from the Microtek with transparency adapter. I will appreciate it very much if anyone of you could tell me whether this could be done or, if yes, how? Thanks in advance. Regards, Yuhui Xu EM Core, DFCI
At 05:39 PM 3/12/98 -0600, Winton Cornell wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have been demonstrating the CD-Rom version of the Particle Atlas extremely successfully for about the last 4-5 months. You can get a copy through McCrone Accessories 850 Pasquinelli Drive Westmont, IL 60559 (800)622-8122 All of our clients (from IBM to Niagara Mohawk) plus students in our recent ACS Course on Applied Optical Microscopy for Chemists have found this a very valuable resource. I think the current price is about $1000 for what is equivalent to an 8 volume, bound set.
Hope this is helpful.
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
Dear Colleagues: I just realized that I can use the positive transparency selection to scan the color slides into the computer. Please disregard my previous posting if this is correct. Thanks. Yuhui Xu EM Core,DFCI
Does anyone know a source for purchasing the simple metal slide type of fiber microtomes? They use a slider to pack the fibers, and then you simply use a razor blade to make a quick and dirty cross-section of the fibers of interest.
I used one over 15 years ago at another company, but no name was on the device.
I appreciate any leads anyone may offer.
THANKS - B.J. CRAVEN E-MAIL to arcraven-at-interpath.com
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------------------------------------------------------------------------------ Post-Doctoral Appointment in Electron Microscopy
A post-doctoral appointment for a person skilled in electron microscopy techniques is open in the Materials and Engineering Sciences Center at Sandia National Laboratories-California. The appointee will apply transmission electron microscopy to the characterization of metallurgical and ceramic materials. The candidate must have received a PhD in Materials Science, Chemistry, or Physics. Experience with diffraction contrast methods, HRTEM, and analytical electron microscopy, including EDS and EELS, is strongly desired. Experience using other microscopy methods, including SEM, is also desirable.
Send resume, with names of references, publication list, statements of research expertise, and copies of transcripts to:
Sandia National Laboratories c/o Anna Isham, MS 9111, HR Dept. -CA0041 P.O. Box 969 Livermore, CA 94551-0969
U.S. Citizenship is normally required. Sandia National Laboratories is an Equal Opportunity Employer/Affirmative Action Employer.
Hi, I'm a Gastroenterology fellow involved in research of bile duct epithelial cells in rats. I would like to stain the intrahepatic bile duct cells in small and larger ducts { 15 u and {15 {300 u in vivo and then obtain a 3D image from a thin slice. Is there a method to apply an antigen to cellular epitopes and subsequently an antibody to the antigen that would have staining properties and would retain some cellular delineation after preparing the histologic slides?. My e-mail address ztretjak-at-usa.net
Hello Everyone, I am attempting to survey how most people identify carbon black. Do you determine N number? Or do you just determine type like ISAF or FEF? Anybody who would like to share their methodology will find an interested party on this end of the wire.
You can reach me at : fskarl-at-goodyear.com or you can send it to the server for everyone. If there is sufficient interest, I will publish a summary of the results.
Hve you tried the folks at Cabot Corp.? I understand that they know a thing or two about carbon black.
I don't know anyone in particular there but there web site is:
http://www.cabot-corp.com
Harold J. Crossman Senior Scientist OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
I am working on the cotton surface free energy analysis. Now I need to apply extreme small liquid droplets to sigle cotton fiber. Anyone has any idea on this? What tool can I use so I can apply the droplet to the fiber automatically?
Since the cotton fiber is extremely fine whose diameter is around 10-15 micron, what kind of syringe will produce small droplet which can stay on the fiber?
This droplet-fiber will be viewd under video microscope and image will be captured using CCD camera for later use.
B. J. Craven wrote: =========================================== Does anyone know a source for purchasing the simple metal slide type of fiber microtomes? They use a slider to pack the fibers, and then you simply use a razor blade to make a quick and dirty cross-section of the fibers of interest.
I used one over 15 years ago at another company, but no name was on the device. ============================================= I think that the microtome you are thinking about if the one featured on our website, specifically URL http://www.2spi.com/spi/catalog/knives/microtome.html
This is called the "Micro Fiber" line of sliding microtomes.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
At 01:14 PM 3/13/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Myles L. Mace, Ph.D. was head of Cabot Corp.'s carbon black laboratory for a number of years. He has done a lot of work with carbon black and would be the best man to help you.
Myles L. Mace, Ph.D. Genetic Microsyatems mmace-at-geneticmicro.com 8 Henshaw Rd. (781) 932-9333 Woburn, MA 01801
Joiner Cartwright, Jr., Ph.D. Assistant Professor of Pathology and Director, Electron Microscopy Laboratory Department of Pathology, Rm.286-A Baylor College of Medicine One Baylor Plaza Houston, Texas 77030 U.S.A. tel.: (713) 798-4658 FAX: (713) 798-3945 joiner-at-bcm.tmc.edu
I think in your case may be caused by the offset voltage of vacuum control circuit is shifted. You need to re-adjust a high vacuum control signal (ready lamp light on). Check at vacuum volage measuring terminal by multimeter after main valve open it should decreasing from about 160uA to 0 uA within few minutes and then the ready lamp will come if not adjust vacuum comparator circuit in vacuum control.
Regards,
Paiboon Nuannin Department of physics, Faculty of Science Prince of Songkla University Hatyai, Thailand
On Thu, 12 Mar 1998, Brian McIntyre wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } hello all- } } i've got a problem with the vacuum system on a JEOL35. the problem seems } to manifest itself by disabling the automatic valve sequencing function. i } can operate all the valves in manual mode but in auto mode it never seems } to reach vacuum ready. in manual mode it pumps OK, but manually operating } the airlock valves is a pain. } } any ideas why manual sequencing the valves works and automatic doesn't?? } } thx } brian } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627 } } 716-275-3058 } 716-244-4936(fax) } } }
The below posting is three days old and has attracted no reply. I cannot = see any purpose why charging would need to be quantified, or how this could b= e properly done. Perhaps the message was misunderstood but as it stands the question is just an interesting exercise to explore. I am not a physicist but happy to give my understanding of the matter. If its wrong, somebody = is sure to respond; I am happy to learn.
In TEM the e beam (probe or beam current) passes through the specimen. Electrons interacting with the specimen have various fates: backscattered and secondary e (as used in SEM or STEM), those flowing through the speci= men to ground (specimen current as measured in analysis), those passing throu= gh the specimen, albeit with diminished energy to form the image.
In SEM, if all emitted e could be trapped and measured in a Faraday cage, that current plus specimen current would equate beam current - except tha= t we have not accounted for any charging. Charging occurs when low energy electrons accumulate on the specimen=92s surface faster than they are conducted to ground. But how could one measure the accumulated electrons = and related that to the other measures. In TEM the situation is further complicated by transmitted electrons. However, charging is not a problem in TEM because the powerful transmitte= d electrons are not affect by the specimen=92s surface charge. The only way= one could notice that a formvar film is charging would be through a secondary detector. If one could technically obtain good measures of all the curren= ts and compare under near identical conditions a number of coated and uncoat= ed films, then one could obtain some measure of electrons trapped on the surface. However, that measure would vary with minor changes, often dramatically. I would like to know what the exercise might achieve. I suspect a set of fairly meaningless numbers. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} Email: knechtew-at-tcd.ie } Name: Walter Knechtel } School: Trinity College Dublin, Ireland } } We are using formvar as a substrate for TEM investigations. } We are studying charging effects on it. Is there any possibility to } estimate quantitativly how much the substrate } charges up when exposed to the electron beam? } The current density of the beam is known. } } Thank you for your help } } ------------------------------------------------------------------------= --- } } }
The PDP11 has been around for a very long time, and is in many respects the model of 'open architecture'. Many third party manufacturers have been producing plug-in boards for it from the beginning. It is quite likely that the TN-2000 controller board was actually OEM'd to Tracor, and not their own design. Frequently the 'sinister' situations I refer to are actually poor communications between the various manufacturer departments - i.e., engineering specs a part to procurement that has only one source, procurement purchases that single part, without paying attention to the package that is actually offered. The incoming parts department is only looking for the specific part and trashes any accessories or software that comes with it. Etc., etc., etc.
Since the controller carries a Tracor part number and might not have any easily identifiable manufacturer ID, the field engineering department assumes that it is a Tracor designed and manufactured part. You are left with that impression and feel that you have no recourse.
The recent migration to PC compatible components has improved the situation somewhat. However, the interface boards required for an EDS system are very esoteric and not generally available. They may well be designed and manufactured by the EDS manufacturer. In certifying a single manufacturer for the computer system however, a manufacturer is once again certifying that it is seeking to lock its customers into an arrangement that is advantageous to the manufacturer. Consider that Link is probably getting substantial consideration from HP for their endorsement.
Frankly, the experience you claim of going ahead and finding solutions that are outside of the manufacturers claims simply endorses my position. If you are willing, there are alternative routes. I'll bet that you were also able to accomplish these changes at a cost substantially less than the manufacturers were offering.
BTW, 3-1/2" drive controllers should be fairly common, look around.
} This came to me directly, and not to the list. You might want to } change the address and post again. } } Personally, I have not run into any "sinister" situations where a } manufacturer deliberately "queered up" just for the sake of } propriety. Our TN-2000 was ostensibly built around a PDP-11 } computer. Yet the state-of-the-art was pretty poor when we bought } it. Tracor had made their own disk controller which didn't use the } full capacity of the disk drives. I think the memory and/or serial } connections may also have been pre-standard. They weren't exactly } non-standard because it was so early in the maturation process. } } We now have a Link ISIS system which is supposedly only checked out } on Hewlett Packard PCs. They supposedly won't guarantee much if we } were to switch to a different brand. That to me is a crock. They may } not certify other platforms, but that doesn't mean they won't work. } If I were more pressed for power, I would upgrade anyway and fully } expect another system to work as well. } } Mostly my experience has been to plug in whatever (on both PDP-11 } and PC platforms) and to make it work. Never mind what the company } says can or can't work. I just need to know what I am doing because } the company may not be any help. Lately, most of the systems have } followed the rules. } } FYI, the outcome of the EDAX 9800 issue was that the system was } built around a PDP-11 and I don't know that the PDP was ever offered } the option of plugging in a 3-1/2" disk. There would definitely be a } few issues to contend with. } } At 04:55 AM 3/11/98 -0600, you wrote: } } The problem actually becomes one of the manufacturers intentionally } } making an open system proprietary. I have no direct experience } } with this particular system, however from my experience, there are } } two possiblities - the manufacturer reps are merely presenting a } } company line and the conversion can actually be made rather easily } } by ignoring their advice, or they are correct and the manufacturer } } has taken steps to ensure that you are locked in to their formats. } } } } Whether it is a windows system or not, manufacturers often make } } hardware and software changes that make it difficult to make your } } own changes. With windows hardware/software you have a cheap and } } easy path to test which way a manufacturer is going. Simply make } } the appropriate changes to the PC hardware and software and see if } } it works. If they are truely programming to windows standards, you } } should be able to change to any hardware you want. However, if } } they are still attempting to maintain proprietary control, your } } changes won't work. In that case, you can always regress to the } } manufacturers setup - and please bitch to them if it doesn't work. } } It may not seem to accomplish much, but if enough people bitch it } } may accomplish something. } } } } } If it is a standard Windows, PC-based system, I see no reason why } } } you could not do that. If your floppy is currently A:, your hard } } } drive is C:, and you have nothing defined as B:, you should be } } } able to do it straightaway. That is the reason for going with } } } standard hardware platforms and operating systems. } } } } } } There is a possibility that the operating environment might be a } } } bit unusual if it is an old PC. It would be more likely that EDAX } } } had somehow customized it or used a non-standard system. It might } } } also be a little tough to get to the CMOS setup program to inform } } } the system what the new hardware configuration is, but you should } } } be able to do it. EDAX may simply be trying to protect itself. } } } } } } Things are much better now than in the old days when expansion } } } was just about impossible. The EDS manufacturers did well with } } } what they had. They sometimes made their own controller cards or } } } wrote their own operating system (I think of my old TN-2000) } } } because standardized, cheap components were not available or } } } because they needed more performance than the standard parts } } } could provide. Matters got better as the PDPs matured and RT-11 } } } and TSX developed as the operating systems - then you could add } } } your own components. But now it is just about a no-brainer. } } } Indeed, I would rather avoid the proprietary systems. They might } } } have a bit of technological edge today, but what will I do } } } tomorrow when I want to slip in a faster processor or a 10 GB } } } disk drive and tape backup? } } } } } } At 08:24 AM 3/6/98 EDT, you wrote: } } } } X-Rayers, } } } } } } } } I have inherited an EDAX 9800 which is attached to an Amray } } } } 1830i. I was wondering if I could replace the 5.25 inch drive } } } } with a 5.25/3.5 inch drive? It makes sense to me, and would be } } } } practical, but the tech support at EDAX said that it would be } } } } impossible and crash the system; which does not make sense to } } } } me.....yet. Are they right at EDAX? Or, are they trying to } } } } get me to upgrade my system? Please enlighten me for I am an } } } } X-Ray novice. } } } } } } } } } } } } John Grazul } } } } Rutgers University } } } } Electron Imaging Facility } } } } } } } } } } } Allen R. Sampson } } Advanced Research Systems } } 317 North 4th. Street } } St. Charles, IL 60174 } } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net } } WWW: http://www.mcs.net/~ars } } Analytical instrument maintenance services } } } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
You are invited to provide information for the microscopist salary survey by return email. After recording your data, your message will be erased. We have NO interest in any individual data. However, a completely "blind" reader respnse card will be included in the April issue of Microscopy Today. This initial survey will relate only to the U.S. and will exclude manufacturers and suppliers. Your data should be include 8 fields, with one entry in each field: Example: PhD-15-M-32,400-N-MW-B-H
FIELD 1: EDUCATION (DEGREE) None/AA/BS/MS/PhD/MD FIELD 2: YEARS MICROSCOPY EXPERIENCE (After Education) ______ FIELD 3: GENDER M - Male F - Female FIELD 4: YEARLY INCOME ___________ FIELD 5: CURRENT SUPERVISOR/MANAGER Y - Yes N - No FIELD 6: LOCATION (If a question, pick the area you feel closest to your own income location) MW - Midwest NE - Northeast SE - Southeast S - South W - West Excluding California CA - California FIELD 7: PRIMARY INTEREST IN: B - Biological Science P - Physical/Material Science E - Earth Science FIELD 8: WORKING IN I - Industry E - Education H - Hospital/Medical G - Government (with GS-Scale) GS - Government Sponsored Research Ctrs
Thank you for your input and help! Don Grimes, Microscopy Today
Does anyone know of any seminars or classes for the instruction of kidney processing for TEM? I am an MT in a hospital lab who would like to learn from an experienced person the proper technique for processing kidney biopsies for diagnostic viewing in a TEM.
I have been asked for cork for microtomy (not ultra microtomy). I have used pith, polysterene, cryo and paraffin wax, but never cork. Does anyone know of protocols for 'cork microtomy' and suppliers of this substance?
Stephan Helfer Royal Botanic Garden Inverleith Row Edinburgh EH3 5LR Scotland UK
} ---------- } Von: Deutschlaender, Norbert, HMR/DE } Gesendet: Montag, 16. M=E4rz 1998 10:57 } An:=20 } From my own experience I fully agree with the statement, that at } present the Olympus BX 40 light microscope seems to meet ergonomic } requirements. Since I have this microscope in use for several years } (many hours per day), I recovered from many shoulder-,neck- and arm } problems - which I did not bear as a "badge of honour" at all -. } Additional to the ergonomic items already mentioned by Malcolm, it } owns the great advantage of indefinitive optics, so that you can } insert as many intermediate rings as necessary to individually adapt } the distance between bench height and position of eyepiece according } to your body size, to maintain an upright position . I have no } commercial interest in any microscope manufacturer. } =20 } Norbert Deutschlaender, Germany (country of Leitz and Zeiss).=20 } ---------- } Von: } = malcolm.haswell-at-sunderland.ac.uk[SMTP:malcolm.haswell-at-sunderland.ac.uk } ] } Gesendet: Donnerstag, 12. M=E4rz 1998 16:45 } An: microscopy-at-Sparc5.Microscopy.Com } Betreff: Re: Hand numbing problems from micro } =20 } = ---------------------------------------------------------------------- } -- } =20 } =20
Years ago I was able to buy a spray-on Teflon coating that when baked would produce a film on glass slides and coverslips. This allowed us to polymerize epoxy embedded cells in an extremely thin film, pick out individual cells for sectioning and pop the coverslip off and remove the epon layer from the slide. In this way we avoided other glass removal steps such as disolving with HF.
This Teflon repair spray does not seem to be available anymore and I have used the last of my coated slides and coverslips. Does anyone know of a comparable "slippery" coating that will allow one to do flat embeddments and then remove the epon layer from the slide?
Mark A. Farmer Director, Ctr. Ultrastructural Research University of Georgia, Athens, GA 30602 (706)542-4080 Voice (706)542-4271 FAX farmer-at-cb.uga.edu http://www.uga.edu/caur
May I suggest a nebulizer, there are many types and they make very small droplets. I dont know the company but I have seen a small glass nebulizer that is operated by a squeeze bulb and works very well. Maybe one of the suppliers on this list even carries this little gem. Alternatively there are home nebulizer kits for drug delivery that should be easy to get your hands on. Good luck, Scott
} } Hi, } } I am working on the cotton surface free energy analysis. Now I need to } apply extreme small liquid droplets to sigle cotton fiber. Anyone has any } idea on this? What tool can I use so I can apply the droplet to the fiber } automatically? } } Since the cotton fiber is extremely fine whose diameter is around 10-15 } micron, what kind of syringe will produce small droplet which can stay on } the fiber? } } This droplet-fiber will be viewd under video microscope and image will be } captured using CCD camera for later use. } } Any suggestions are highly appreciated! } } } Jing Lu }
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
At 10:03 AM 3/16/98 GMT, Stephan Helfer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have routinely used cork as a substrate to support a bundle of fibers for fiber microtomy, but have simply used a routine laboratory cork. We've cut about a 1/2 (100 micron) cross section, used a large eyed sewing needle to pull a collection of fibers through the cork, then used a new, single-edged razor blade to microtome cross sections of the fibers. This approach works especially well since the cork is opaque, making the fiber bundle easy to find.
Along similar lines, I a rather unusual situation when no cork was present, I once cut about an inch off the top of a good sized carrot, cut the carrot in half longitudinally, then mounted a piece of fabric in the cut and bound the carrot/fabric sandwich back together with sewing thread (this was really a case of Necessity being the Mother of Invention!), then microtoming cross-sections of the fabric, again, with a single-edged razor. This procedure took a little practice, but within a few cuts, we were getting sections credible enough to be used in a court case. A cork would have worked just as well.
Since you are working for a botanical garden, perhaps you can comment on different type of corks. What, for example, is typically used for the standard laboratory corks? Is there a difference in cork density determined by which plant it comes from? It would seem a though there needs to be a good balance between enough density to hold a sample in place versus being too dense to cut easily with whatever type of blade is used for microtoming.
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
Dear Microscopists, I would be very glad if anyone could point me to papers with good description of yeast ultrastructure (strains S.cerevisiae and S.pombe). Thank you in advance.
To Microscopy, Is it possible to do the silver-enhanced (1nm gold) technique on parlodion or pure carbon grids to small particles and then negative stain. The real question is are any of the silver-enhancement chemicals detrimental to a grid destined for negative staining? Thanks
I recently read about a compact field microscope with simple 35mm attachment, called the Lensman. I am looking for all info that I can get about it, since the article, which I have seen in one of Hedgecoe's general photography manuals, did not tell anything about when or where it was manufactured or by whom. Or I would be interested in being pointed toward information on similar microscopes which allow photomicrography to be performed in the field.
Thanks, Doug Darnowski
****************************************************************************** Douglas Darnowski Department of Crop Sciences 384 ERML 1201 West Gregory Drive University of Illinois Urbana IL 61801 work ph: (217) 244-6150 fx: (217) 333-4777 home ph: (217) 356-6606 fx: (217) 356-4454 email: darnowsk-at-staff.uiuc.edu
} Years ago I was able to buy a spray-on Teflon coating that when baked } would produce a film on glass slides and coverslips. This allowed } us to polymerize epoxy embedded cells in an extremely thin film, pick } out individual cells for sectioning and pop the coverslip off and } remove the epon layer from the slide. In this way we avoided other } glass removal steps such as disolving with HF. } } This Teflon repair spray does not seem to be available anymore and I } have used the last of my coated slides and coverslips. Does anyone } know of a comparable "slippery" coating that will allow one to do } flat embeddments and then remove the epon layer from the slide?
When we flat embed, we use Aclar plastic sheets cut to whatever size we want with a pair of scissors. Cells can be grown on the Aclar. Some types of cells have no trouble sticking but some need the aclar to be coated with poly-l-lysine or glow discharging to help them attach. After polymerisation, the Aclar peels away leaving the cells in the block or if the plastic is left on, it can be cut with a glass or diamond knife. If it is straight flat embedding, two pieces of Aclar are used with the specimen sandwiched between. After polymerisation, one piece of aclar is peeled away, the specimen is glued to a form block and the other piece is peeled away. Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
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We have an older Kevex X-ray analysis system which would be available to anyone willing to pay shipping charges. This Kevex model 8002 microanalyzer system was designed for a JEOL JSM-840 scanning electron microscope. It has a light element x-ray detector and was used primarily on biological samples. The entire system including detector, 2 ion pumps, motorized system for moving the detector into the column, and computer console, boards, etc. is available. The detector was just removed from the microscope a few months ago and is presumed to be in good condition although we suspect that the preamplifier is defective. This could make a good unit to have for parts if someone has a similar unit they are still using. If you are interested in this equipment, please contact:
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Purdue University E-mail: sherman-at-aux.btny.purdue.edu W. Lafayette, IN or: emcenter-at-btny.purdue.edu
(InterLock SMTP Gateway 3.0 for microscopy-at-Sparc5.Microscopy.Com); Mon, 16 Mar 1998 14:15:11 -0500 Received: by na3.dow.com (Internal Mail Agent-1); Mon, 16 Mar 1998 14:15:11 -0500 Message-Id: {c=WW%a=ATTMAIL%p=DOW%l=MANTE09-980316191502Z-96227-at-mante02.nam.dow.com}
Greetings:
We have been experiencing what could be an anomalous B peak detected by our PC3 (MoB4C) WDS crystal, particularly when analyzing C based samples. Has anyone had problems with secondary fluorescence of B in the crystal generated by C K-alpha emissions. Could such an event yield a small B peak, or would the effect be apparent as a higher than usual background signal?
We are very interested in resolving this issue. If secondary fluorescence of B would not generate a small peak, we may have contamination of our standard block from previous B analyses.
Any suggestions or experience regarding this phenomena would be highly appreciated.
Regards
Cary Black Dow Chemical phone: (517) 636-5760 e-mail: ckblack-at-dow.com
Mark, I use Permanox slides from EMS (I bet other vendors carry them as well) for flat embedding. I have some if you'd like to try them. No spray is needed and the slides separate easily.
your neighbor, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
One the of fluorescence filter cubes on my Zeiss Axiophot just fell apart! Apparently the repeated heating cooling cycles caused the thin threaded ring that held the excitation filter in place to loosen. The ring and the actual glass excitation filter fell out. Fortunately nothing broke and it was easy to re-assemble. Can anyone confirm that there is no "sideness" to the excitation filter (i.e., that it doesn't matter which direction I re-inserted it)? Thanks in advance - Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Mark Farmer wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Years ago I was able to buy a spray-on Teflon coating that when baked } would produce a film on glass slides and coverslips. This allowed } us to polymerize epoxy embedded cells in an extremely thin film, pick } out individual cells for sectioning and pop the coverslip off and } remove the epon layer from the slide. In this way we avoided other } glass removal steps such as disolving with HF. } } This Teflon repair spray does not seem to be available anymore and I } have used the last of my coated slides and coverslips. Does anyone } know of a comparable "slippery" coating that will allow one to do } flat embeddments and then remove the epon layer from the slide? } } Mark A. Farmer } Director, Ctr. Ultrastructural Research } University of Georgia, Athens, GA 30602 } (706)542-4080 Voice (706)542-4271 FAX } farmer-at-cb.uga.edu } http://www.uga.edu/caur
Dear Mark,
We at Ladd Research sell a teflon spray you may be interested in. Our catalog number 21845. If you are interested I can send you pricing and technical data.
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Check out the MAS Homepage for the latest information on the Microscopy and Microanalysis Meeting in Atlanta this year. http://www.micoranalysis.org I have just added the Schedule of Scientific Presentations. Complaints on errors and omission to the chair please! :-) (Kathi Alexander alexanderkb-at-ornl.gov)
Jfm. Actual full path to the schedule is: http://www.microanalysis.org/mas/masmti/m&m98/schedule.html
________________________ Note new Area Code (734) ________________________ John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Last month I asked for current opinions on high-quality printers for {$10,000. Our choices included getting an Epson Sylus Color 800 ( {$400) and a Fargo dye-sub printer (~$2,000), or just a more expensive dye-sub printer (e.g., Tek 450 for $8,000).
Pretty much everyone said find a couple of thousand dollars more and get the Codonics NP-1600 dye-sub printer.
We performed a casual test of several printers and came to the folowing conclusions:
Ink jet technology will satisfy more than 80% of the color printing needs that our group (mostly neurobiologists with some behavioral psychologists and morphologists thrown in) will have. Our LANlord has ordered the network versions of the Epson Stylus 800 and the Epson Stylus Photo. These printers do a very good job of mixing text and photos (which the dye-subs do not), and their costs of operation are very reasonable.
Of the dye-sub printers, the Kodak and Codonics produce the best color and b&w images (although Fargo has not yet sent printouts of the file we sent them). The Codonics has the Kodak print engine, but has nicer software and networks much more readily than the Kodak. It also produces good color transparencies.
When faced with the question of why get such an expensive "digital darkroom" when a combination of ink jet and laser jet printers might take care of most of our needs for far less money, we were reminded of this: Most labs or individuals can afford something like the Epson 800, but no one lab can afford the Codonics. It makes sense to put the money towards that for our multi-user environment, and let individuals buy their own ink jets or laser jets.
Thanks for all your opinions and experiences!
Aloha, Tina
80F, sunny, dry, surf's up. Can't afford the housing, but who cares?
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
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Doug -
What kind of magnification do you need? I use a nice screwon front-of-the-lens auxiliary from Edmund that gives a reasonably well-corrected 8x, unlike simple diopter lenses.
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
I have always use Teflon Spray made by Miller-Stephenson. # MS122-22, TFE Release Agent, Dry Lubricant. It cost $10.50 per can of 455gm the last time I purchased it. The company has offices in Ontario, Conneticut, Illinois & Los Angeles. The LA # is 203-743-4447. Another number is 818-896-4714.
Dear All, I need some antibodies against matrix metalloproteinases that, according to literature, should be available from a company called Oncologix, Gaithersburg and/or Molecular Oncology, Gaithersburg. I was not able to find these companies in Linscott's Directory of antibodies nor on the Net. Could somebody give me their addresses, phones or faxes, please? Thanks for your help,
Sarka Lhotak
Hamilton Regional Cancer Centre McMaster University Hamilton, Ontario, Canada
Tom, With Omega optical filters the light must pass through the filter in a particular direction, and it is likely that this is the case with your filter as well. Usually, there are arrows on the filter ring indicating which direction the light must travel through the filter.
Miss Peta Clode Zoology Department LaTrobe University Bundoora, Victoria Australia. 3083. Ph (03) 9479 2177 / 2279 Fax (03) 9479 1551
On Mon, 16 Mar 1998, Tom Phillips wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } One the of fluorescence filter cubes on my Zeiss Axiophot just fell apart! } Apparently the repeated heating cooling cycles caused the thin threaded } ring that held the excitation filter in place to loosen. The ring and the } actual glass excitation filter fell out. Fortunately nothing broke and it } was easy to re-assemble. Can anyone confirm that there is no "sideness" to } the excitation filter (i.e., that it doesn't matter which direction I } re-inserted it)? Thanks in advance - Tom } } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211 } (573)-882-4712 (voice) } (573)-882-0123 (fax) } } }
If I understand correctly you actually have two questions: 1. What does a (filmed) grid do with silver enhancement? 2. Does silver enhancement give problems with negative staining of for instance immunolabeled silver enhanced (virus) particles?
Ad 1. We always recommend to use nickel grids with parlodion or formvar for enhancement. I am sure carbon grids will be fine. Copper can give a lot of dirt, since this element gets involved in the chemistry that will be going on. Don't use gold grids, unless you want silver grids afterwards.
Ad 2. This is a more difficult question to answer and I am afraid my answer will be lengthy. First of all, since you are using ultra small particles, you will get the best results with the silver enhancement procedure according to Danscher or a method that is derived from this one. You will find a description in: Histochemistry 71, 1981, pp 81-88. Many commercially available (more or less neutral pH) silver enhancement reagents (such as ours) have been developed for general applications for instance in light microscopy. In order to obtain a homogeneous, time controlable and efficient enhancement of ultra small gold particles you have to add Gum Arabic to the mix of components. However, comparison of the performance of many enhancement reagents shows that Gorm Danscher's method is still the method of choice. Secondly, fragile and unfixed specimens may suffer from the action of chemicals in the reagents, after all there are redox reactions involved. The pH may play a role and you may have osmotic effects, although if you're planning an acidic negative stain afterwards I would not worry too much about that. Neither about osmotic problems since while the negative staining solution dries on your grid its osmolarity will rise to extreme levels. To prevent ultrastructural changes at least to some degree I suggest to use a fixation after the immunoincubation steps, but before silver enhancement. Third, substances like Gum Arabic or high molecular weight additives, present in some reagents, are very viscous and sticky and thus difficult to completely remove from your grid and particles. I suspect that this might prove to be the biggest problem as residues of these substances will reduce resolution. Therefore you will need prolonged and extensive washing after the enhancement step and before negative staining.
And finally, if you have the possibility to use High-Angle Annular Dark-Field STEM Imaging you don't need the silver enhancement. This visualization technique allows you to observe unenhanced ultra small particles in unstained very thin specimens. Reference: Otten et al. SCANNING Vol. 14, 1992, pp 282-289.
I hope this answers your questions to a certain extent. Feel free to contact our HELPDESK via our web site at http://www.aurion.nl if you need more detailed information.
============================= Jan Leunissen, Ph.D. AURION ImmunoGold Reagents & Accessories Managing Director Costerweg 5, 6702 AA Wageningen The Netherlands
phone (31)-317-497676 fax (31)-317-415955
please visit us at the AURION Website: http://www.aurion.nl/
I embed vibratome sections as slides and then remove areas of interest for re-embedding. I use a Teflon Release Spray for coating the slides. It is available from RS Components in the UK. It works very well. There may be an equivalent available in the USA. You just spray it on the slides, polish gently with a cloth and that is all. No baking needed.
You could also try using Silicon coating using Sigmacoat. The solvent is not nice but used with care it works very well for releasing coverslips from "embedded" sections on slides. I just dip the coverslips in the liquid allow to air dry and use them straight away.
The coverslips usually lift off the section very easily and the section or any part of it is very easily released from the slide.
---------- } From: Mark Farmer {farmer-at-emlab.cb.uga.edu} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Flat Embedding } Date: 16 March 1998 10:24 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Years ago I was able to buy a spray-on Teflon coating that when baked } would produce a film on glass slides and coverslips. This allowed } us to polymerize epoxy embedded cells in an extremely thin film, pick } out individual cells for sectioning and pop the coverslip off and } remove the epon layer from the slide. In this way we avoided other } glass removal steps such as disolving with HF. } } This Teflon repair spray does not seem to be available anymore and I } have used the last of my coated slides and coverslips. Does anyone } know of a comparable "slippery" coating that will allow one to do } flat embeddments and then remove the epon layer from the slide? } } Mark A. Farmer } Director, Ctr. Ultrastructural Research } University of Georgia, Athens, GA 30602 } (706)542-4080 Voice (706)542-4271 FAX } farmer-at-cb.uga.edu } http://www.uga.edu/caur
Does anybody have access to an SEM with a stage capable of greater than 1800C? If not, then how about a system that can handle 1400C? We might need to run a few high temperature experiments, but not enough to purchase a system.
Please contact me off-line if you have such a tool.
Thanks in advance.
Harold J. Crossman Senior Scientist OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
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RE} fiber microtome 3/17/98 8:50 AM Dear all,
The SPI slide microtomes are very good, yet expensive. You may also be in= terested in doing the sectioning by hand using Jollif Plates. These may m= ake too thick of sections for you, but they may work for what you want.
The Jollif Plates can be had from McCrone Associates (with the technique)= for pennies a piece.
McCrone Associates can be contacted at 800-622-8122.
John Shane McCrone Research Institute (not affiliated with McCrone Associates)
--------------------------------------
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
B. J. Craven wrote: =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D Does anyone know a source for purchasing the simple metal slide type of fiber microtomes? They use a slider to pack the fibers, and then you sim= ply use a razor blade to make a quick and dirty cross-section of the fibers o= f interest.
I used one over 15 years ago at another company, but no name was on the device. =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D I think that the microtome you are thinking about if the one featured on = our website, specifically URL http://www.2spi.com/spi/catalog/knives/microtome.html
This is called the "Micro Fiber" line of sliding microtomes.
Chuck
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
for Microscopy-at-sparc5.microscopy.com id AA08073; Tue, 17 Mar 98 16:46:00 +0100 Message-Id: {9803171546.AA08073-at-iris1.fae.ub.es} To: Microscopy-at-Sparc5.Microscopy.Com, Microscopy.com
Dear All
To members of EU: Two post-doc positions are offered for graduate students or PhD. The objectives are the structural and compositional characterization of UV-coatings. Is someone is interested, please contact Dr. F. Peir=F3 at
paqui-at-iris1.fae.ub.es
Please find more information visiting the web site: www.lzh.de/tmr (Please remark the change of the web site address with respect to a previous message).
by UTSW.SWMED.EDU (PMDF V5.0-7 #18231) id {01IURS56UAXY8Y4X4A-at-UTSW.SWMED.EDU} for Microscopy-at-MSA.Microscopy.Com; Tue, 17 Mar 1998 11:30:44 -0500 (CDT)
Hi, This is a "case of emergency". I would like to do Quick freeze-deep etch technique in our lab to study the shape of a protein we purified. We have everything to start except the copper block that would allow me to freeze my samples. We have a slammer designed by Tom Heavy.I am looking for a copper block. I am planning to use nitrogen to do so. So, if you have an extra copper block to lend or to give or sell. Please, tell me.
Thanks.
Val=E9rie Nicolas The University of Texas Southwestern Medical Center at Dallas. 5323 Harry Hines Blvd. Dallas,Tx 75235-9039 Phone # (214) 648-3657 Fax # (214) 648-9160
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The State Microscopical Society of IL (SMSI) is having a meeting this Friday evening in Chicago with the speaker Ankur Purohit of Argonne, speaking about "Diamonds: Man's Best Friend". The meeting starts at 6 p.m. with pizza( $6 ) and the talk is at 7 p.m. it is held at McCrone Research Institute, 2820 S. Michigan Ave. and you need to call if you want pizza (312-842-7100). There will be some new information on doped diamonds and commercial opportunities.
I have a JEOL T300 scanning microscope. Last year i had arching problem in the HT cable at the gun end. I was able to cut the portion where the arching was and rerouted the cable to have enough length. Things worked fine since then. However, I had a similar problem last week in almost the same place. Unfortunately this time I have no extra length to shorten the cable which left me with one solution. To replace the cable. I contacted JEOL and i was informed that they don't sell the cable separately but a cable and a gun together. The cost around $3000. I am not in a position to spend that much money anf the gun works fine. I would like to know if any body has any idea on a supplier where I could buy a HT cable or if you know someone having an old T300 or any other ideas.
Thank you
said
== Said A. Mansour == Purdue University == School of Materials Engineering == 1289 MSEE Bldg. == W. Lafayette, IN 47907-1289 == # (765) 494-6405 Fax (765) 494-1204
Does anyone recall the procedure for making molybdenum trioxide crystals on a TEM grid. They will be used for calibrating the rotation between the image and diffraction pattern. Tia-Mike
The easiest way to do it is to use carbon coated grids and an evaporator with a Mo boat. (you can also use Mo wire if you like) With the evaporator bell jar off and the system up to air, heat the filament until you get white smoke rising. Then simply wave the carbon coated grid in the smoke a few times and you have your sample.
If you are doing this for a rotation calibration sample, you might consider using John McCaffrey's Mag-I-Cal sample available from South Bay Technology instead. It can be used to for mag calibration from about 1000X up to your machine's highest mag, rotation calibration, and camera constant all-in-one little sample. I've used it and it works very well.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Does anyone recall the procedure for making molybdenum trioxide crystals on a TEM grid. They will be used for calibrating the rotation between the image and diffraction pattern. Tia-Mike
My friend has a question for you. If we have a ball consisting of spherical shells sharing the same spherical center. If we have enough number of shells and the distances between shells are the same, e.g., 30A. What does its electron diffraction look like? Is that true that the "lattice" fringes look like circles with the same center?
I know this is an optimistic request but ! If anyone has a Kevex 4505P pulse processor module , [ originally part of a Kevex 7000 E.D.S. system ] , they no longer use or need , could you please contact me directly to negotiate a sale price . Thanks Michael Griffiths griffiths.michael.mj-at-bhp.com.au
B.H.P. Steel , Newcastle , New South Wales , Australia
I am embedding cellulose fibers in an Epoxy resin mixture. The mixture is cured for 48 hours at 60C. It appears to shrink and put the sample under compressive forces because after cutting, the slices contains waves and want to expand. The problem I am having is that the cellulose will not allow the resin to relax after cutting, thus the resin tears the cellulose fibers apart.
Does anyone have any suggestions for other low viscosity resins that will not shrink upon curing or general ideas for what I working with?
} } Does anyone recall the procedure for making molybdenum trioxide crystals on } a TEM grid. They will be used for calibrating the rotation between the } image and diffraction pattern. } Tia-Mike } }
you just need a voltage variator to which you connect a piece of molybdenum wire. heat the wire slowly until it fumes. Have a grid with carbon film ready to pass through the smoke, and small MoO3 crystals will deposit on it. Actually I am planning to make a few of them this afternoon...
Yves MANIETTE Universitat de Barcelona Serveis Cientifico tecnics Carrer Lluis Sole i Sabaris, s/n E-08028 BARCELONA Tel +34 3 402 1695 Fax +34 3 402 1398 http://www2.gol.com/users/scscope/maniette/ENTREE.HTM
We have a low tech approach to making moly trioxide crystals that is similar to Scott Walck's. Heat ammonium molybdate in a beaker on a hotplate until white smoke appears, then wave a carbon filmed grid in the smoke. Be sure to do this in a chemical fume hood.
Joe Neilly Microscopy and Microanalysis Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-3537 Voice: 847-938-5024
You can use ammonium molybdate and heat this in a crucible (using a burner) until it becomes red hot. At this point it will sublime. The crystals which will collect near the top of the crucible can then be washed with water (or methanol) to form a dispersion. A drop of the dispersion can then be deposited onto a carbon coated grid. The whole process takes only a few minutes. Make sure you work under a hood. (Ref. Hirsch et.al.)
Jordi Marti.
Does anyone recall the procedure for making molybdenum trioxide crystals on a TEM grid. They will be used for calibrating the rotation between the image and diffraction pattern. Tia-Mike
Greetings, Robert Dickson asked: } } Does anyone have any suggestions for other low viscosity resins that } will not shrink upon curing or general ideas for what I working with?
We work with a mixture of 4:1 butyl:methyl methacrylate, polymerized at 4C with UV light (365nm). We have measured the sizes of plant roots before and after embedding and found no change (to within about 1-2%). With really large block faces (i.e., 6mm x 1mm), we have measured some compression on sectioning, but not with smaller ones. In the development of "lowicryl" resins for low temperature embedding, Carlemalm et al. 1982 J. Microsc. 126:123-143 also found that a very similar type of methacrylate mixture did not shrink with polymerization.
If this sounds as though it might be useful, and you have further questions, please let me know. Tobias Baskin
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Position Summary:
Unilever Research, based in Northern New Jersey, is seeking highly motivated candidates for a position in Electron Microscopy.
Job Description:
1. Assist and if needed perform tissue specimen preparation, prepare cryo samples for freeze fracture and for transmission electron microscopy of frozen-hydrated samples. 2. Prepare freeze-fracture replicas from frozen samples. 3. Conduct transmission electron microscope work of prepared specimens under general supervision. 4. Assist in maintenance and repair of equipment and laboratory 5. Assist and perform image handling functions (digitize, process, measure, label and print images). Maintain imaging equipment (digitizer, computer, printer and other ancillary equipment). 6. If needed, teach and assist users in proper operation of equipments. 7. Perform administrative duties such as data entry, billing, ordering, record maintenance etc. 8. Conduct critical literature evaluations of science and technologies related to research projects. 9. Contribute to data documentation and publication of articles. 10. Train junior staff in proper and safe lab skills and instrument use.
Qualifications:
1. Masters degree in a field appropriate to the area of assignment AND three years relevant research experience in microscopy and cryo techniques; OR,
2. Bachelors degree and six years relevant research experience in microscopy and cryo techniques.
As an international leader, Unilever offers a competitive salary, comprehensive benefits, unique challenges and the opportunity to share your ideas with some of the industry's brightest professionals. Please forward your resume to Dr. M. Misra, Unilever Research, 45 River Road, Edgewater, NJ 07020. Unilever is an Equal Opportunity Employer m/f/d/v.
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Hi all, Dito on the message below, plus is there a Microcroscopy Society group that meets in Dallas?
Karen Pawlowski Dept. of Otolaryngology UT Southwestern Medical Ctr. Dallas, TX
On Tue, 17 Mar 1998, George Lawton wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I would appreciate getting any information on the next Texas Electron } Microscopy meeting. } } thanks, } } George W. Lawton } Cell Biology and Neuroscience } U.T. Southwestern Medical School at Dallas } Phone: (214)- 648-7291 } Fax: (214)- 648-8694 }
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hi, there,
My friend has a question for you. If we have a ball consisting of spherical shells sharing the same spherical center. If we have enough number of shells and the distances between shells are the same, e.g., 30A. What does its electron diffraction look like? Is that true that the "lattice" fringes look like circles with the same center?
(InterLock SMTP Gateway 3.0 for microscopy-at-sparc5.microscopy.com); Wed, 18 Mar 1998 11:08:37 -0600 Message-Id: {199803181708.AA19753-at-relay.ppco.com} Received: by relay.ppco.com (Internal Mail Agent-1); Wed, 18 Mar 1998 11:08:37 -0600 X-Sender: mdphill-at-pop3-host.ppco.com X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.5 (32)
The Oklahoma Microscopy Society & The Texas Society for Electron Microscopy Will present: April 2-4, 1998
"Back to the Future: Improving Your Image"
at:
Lake Texoma Resort (on the Texas-Oklahoma border)
STYLE:Invited MSA, MAS and Outside Guest Speakers, Workshops, Platform & Poster Presentations, Exhibits, and Microscope Demonstrations
LOCATION:Lake Texoma Resort: "A Scenic Water Playground" Phone: (405) 564-2311 or (800) 654-8240 and (select 2) Activities: Excellent fishing, Boat Rental through Catfish Bay Marina, Challenging Golf, Recreation and Aquatics Nature Centers Mailing address: Box 248, Kingston, OK 73439
AGENDA
Thursday April 2, 1998
12 noon - 7pm Registration 12 noon - 7pm Exhibitor setup & Poster setup 5 - 7pm Executive Council Meetings: Officers for Oklahoma Microscopy Society and Texas Society for Electron Microscopy 7 - 9pm **EVENING SOCIAL**: Snacks and Beverages
Friday April 3, 1998
7:30 - 8:30am Continental Breakfast 7:30 - 5pm Commercial Exhibits 7:30 - 5pm Poster Presentations 8am - 5pm Registration MORNING PLATFORM SESSION 8:30 - 9:30am Keynote Speaker: Dr. Jack Kinnamon, Preliminary Title: Digital Manipulation of Acquired Images: What is Possible vs. What is Ethical? 9:30 - 10am Platform Presentations 10am - 10:30am Break 10:30am - 11am Platform Presentations 11 - 12 noon Keynote Speaker: Dr. Greg Meeker, Preliminary title: Standards for X-ray Microanalysis: How We Get Them,How We Use Them and Why We Will Always Need Them. AFTERNOON/EVENING SESSION 12:30 - 5:pm Workshops (preregistration/fee required), Posters and Exhibits, Question and Answer Session 6 - 8pm Dinner 8 - 10pm After Dinner Presentations, Dr. Ben Powell and Dr. Charlotte Ownby
Saturday April 4, 1998
7:30 - 8:30am Continental Breakfast 7:30 - 12 noon Commercial Exhibits 7:30 - 12 noon Poster Presentations 8am - 10pm Registration MORNING PLATFORM SESSION 8:30 - 9:30am Keynote Speaker: Dr. Ken Moore, University of Iowa, Title: Correlative Microscopy (LM, Confocal, SEM, and TEM) of Ocular Melanoma, Pathogenic Bacteria, Cystic Fibrosis, and Other Human Diseases. 9:30 - 10am Platform Presentations 10am - 10:30am Break 10:30am - 11:15am Platform Presentations 11:15 - 12 noon Keynote Speaker: Paige Johnson, Conoco, Ponca City, OK, Title: Educational Outreach: How-tos for Classroom Presentations and Lab Tours.
FEES:
Room Reservations: Call the Lake Texoma Resort DIRECTLY Specify you are attending the joint meeting to receive discounted rates! Phone: (405) 564-2311 or (800) 654-8240 and (select 2) When: Make your room reservation early! At least 6 weeks in advance if possible. Room availability: Reserved Rooms: Double-sized bed and 1 twin bed or 1 Queen only (*Other rooms and rates available upon request: cabins and suites) $50 on Thursday April 2, $53 on Friday April 3 (& Saturday night April 4)
Meeting Registration: Deadline March 13, 1998 (Friday the 13th!) (Includes all meals and materials EXCEPT the Friday, April 3 Chuck Wagon Dinner.) Member: $35.00 Non-Member: $45.00 Student: $10.00 Exhibitor: $65.00 Onsite: Add an additional $10.00
Workshop(s): $10.00 per workshop Will be posted on the OMS web site as soon as topics are finalized. http://www.ou.edu/research/electron/oms/
Optional Friday Night Dinner: Chuck Wagon Style: $15.00 Meal Includes: ribeye steaks, baked potato, tossed salad, watermelon boats, marshmallow roast, apple cobbler and your choice of coffee or tea (If inclement weather, an inside banquet will be held instead.) After Dinner: Tentative: Return to lodge to hear from an Invited Speaker For more information, contact: Secretary/Treasurer: Ginger Baker, Em Lab, Research Department, OCOM, 1111 W. 17th Street, Tulsa, OK, 74107 (918)561-8232 e-mail: lizard-at-ok way.ok state.edu
http://www.ou.edu/research/electron/oms/wkshp98.html Updated: 04-Feb-98 Dean Phillips Phillips Petroleum Company Phillips Research Center, Room 360 PL Bartlesville, OK 74004
I would like to know this myself. Regards, ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-480-6925 MSP Failure Analyst, DDAO Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I am trying to locate a supplier for cedarwood oil. We use it as a clearing agent for peripheral nerve fiber teasing. We have been using a bottle of "Oil of Cedar" (which is probably a 100 years old!) which contains the label "Turtox" and "General Biological Supply House, Inc., Chicago, Ill.". Since that supply ran out 6 months ago, I have purchased cedarwood oil from two popular chemical suppliers. Cedarwood oil from one vendor was actually cedarwood immersion oil and turned out to be too thick to perform the fiber teasing procedure. The other bottle we purchased (we actually purchased 3 different bottles over the 4 month period) crystallizes at room temperature, once it is poured into the vials containing tissue. In fact, that cedarwood oil also crystallizes in the bottom of the primary container once it has been opened. I performed an internet search for General Biological Supply House, but no luck.
Can anyone help me?? Thank you.
Sandy Perkins
Laboratory for Neurotoxicity Studies VMRCVM Virginia Tech
I'm passing along a request for information from a colleague concerning microscopy of afferent and efferent fibers of peripheral neurons. She's looking for the most appropriate microscopy method and possible specimen preparation protocols. Any takers?
Thanks.
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
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} } I would appreciate getting any information on the next Texas Electron } } Microscopy meeting.
} } George W. Lawton
I would like to know this myself. Regards, ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford
Don't you Texans talk to each other? There's an up-to-date list of all local societies on the MSA web page; you can contact the officers of ANY local society without using the listserver.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
} I'm passing along a request for information from a colleague concerning } microscopy of afferent and efferent fibers of peripheral neurons. She's } looking for the most appropriate microscopy method and possible specimen } preparation protocols. Any takers?
Sure but you (or your colleague) needs to be more specific about what you want to see. There's a myriad of procedures.
Electron Microscopy Sciences (1-800-523-5874) offers two different Cedar Wood Oils, one for 'cleaning microscope sections', the other 'used as an immersion oil'. Carolina Biological Supply (1-800-227-1150) has one type for both 'clearing and immersion'.
I used the following method quite often: - connect a Mo wire about 5 cm in length and 0,2 mm diameter to a few amps power supply through a cable and clips. - hold some EM grids carbon or formvar coated about 5 to 10 cm above the wire - heat the wire by slowly increasing the current in the Mo wire until it burns and breakes. You will find the characteristic MoO3 crystals, sometimes with a second oxyde forming round particles.
You can also burn a thin Mo basket in your favourite coating unit without closing it (i.e. under room atmosphere) if the power supply is not protected by a vacuum lock.
Easy isn't it? Philippe Buffat
} From: mcalabrese-at-rsc.rockwell.com } X-Lotus-Fromdomain: ROCKWELL } To: microscopy-at-Sparc5.Microscopy.Com } Date: Tue, 17 Mar 1998 14:52:24 -0800 } Subject: Diffraction pattern rotation } Mime-Version: 1.0 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
For fiber teasing in neurotoxicology we use glycerol, followed by glycerol-gelatine (KAISER's) for enclosing. Some people use a low viscosity epon mixture, but this seems heroic, since you have your nose directly over the slide for a long time.=20 Norbert Deutschl=E4nder, Hoechst Germany=20
As we are entering the realm of image analysis and quantification, we are dealing more frequently with our statistician in sampling designs and data handling related to our microscopy. We have him keenly interested in image analysis and the statistics associated with it.
At the same time, we are also looking for the "perfect" image analysis software. Our statistician has done his own searching. From his vantage point, he is very supportive of the Iris Explorer software. We have had access to a demo to try out, which can do image analysis and apparently has strong data handling capabilities.
I sat down with it and went throught the tutorials and other demos, and tried to imagine how I would use it. From the time I spent with it, it seemed to me that it could make images from data (number) files, but I couldn't see how it worked the other way around (images in a graphics file quantified). How would images be imported into the software?
Could anyone explain a little more about how a microscopist could use this software? As I said, the statistician is keen to get it and has asked for my opinion and support.
Thanks in advance. Please contact me offline - if a sufficient number of people are interested I will post a summary here.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia B4N 1J5
If you can use essential oils ( as in aromatherapy or perfume making) a good source is Janca's Jojoba Oil Co. in Mesa, Arizona. The email address is: JANCAS3-at-aol.com I have the phone and address at home and can bring tomorrow.
Julie Gross UCONN Health Center Farmington,CT 06029 860-679-2463 jgross-at-neuron.uchc.edu
For preparation of single fibers in neurotox we use glycerol on glass-slides, followed by glycerol-gelatine (KAISER's) for enclosing. Some people dissect the fibers in low viscosity EPON, but this seems heroic, since you hardly can avoid exposing your nose for a longer time to the vapor of monomers. Norbert Deutschl=E4nder, Hoechst Germany=20
The issue of ethical digital image manipulation was discussed at previous MSA-MAS meetings but I still have questions.
What is appropriate manipulation of microscopy images (particularly grey-scale TEM and SEM) for scientific and public relations purposes? How acceptable is pseudo-colorization even for public relations? Are painting programs as appropriate for this purpose as the scientific packages that use grey-scale to color manipulation for the whole image? Is discretionary painting of parts of the image as acceptable as the placement of an arrow to highlight specific parts of an image? For public relations purposes with non-microscopist scientists and others, how much information is needed to explain the digital manipulations used?
There are other questions that may be asked but this is a start for discussion.
On Thu, 19 Mar 1998, michael shaffer wrote: } } What is appropriate is relative to (... in my mind ...) appropriately } presenting the image's history of manipulation. I believe any researcher } should be able to manipulate for "clear" presentation of the point he/she is } making ... AS LONG AS, how the image was manipulated is also evident. } He/she should be able to convey the confidence in the presentation and that } it is not presenting misinformation. } For example, before modifying brightness, contrast, gamma, or } superimposing false color, I generally paste a strip which represents the } original "rainbow" of grays or color along with the image. Any manipulation } after that will be presented in this strip and should be redily evident. } This is a good representation of modifying the histogram, but I have yet to } come up with something which would represent manipulations like noise } removal ... e.g., the size of a median kernal or whether it was a gaussian } averaging filter ... etc ... Ideas?? } } cheerios, shAf }
But that really begs the question, or rather, ignores the question. *Any* image that is digitally acquired or printed is "manipulated." That's what color management *is,* for instance.
Consider the following: Let's say that I acquire an image using a digital camera attached to a microscope. It turns out that the colors the camera can aquire (the gamut) is likely *different* than the colors the monitor I look at can display. What I can display varies with the contrast, gamma, etc. of both devices. Now I want to print it. Guess what -- the gamut of the printer is likely different than the gamut of either the monitor or the camera. You want to know all the changes that are made at each stage? Does that mean that you want me to publish all the color management tables, all the calibration maps, all the impulse reponse curves?
Of course not. Nobody wants that data. So, if contrast is manipulated "behind the scenes" as part of the basic data aquisition and color management, we don't need to describe it. If we do it manually we do? That doesn't make sense -- if I assume that some table written in the software is optimized for my particular monitor or printer, that's OK, but if I modify that table so that it actually *is* optimized I have somehow "manipulated" the image?B
Of course, we are mostly completely unaware of all the manipulation and mapping that goes on to get from one place to another, but the fact is that manipulation for "clear" presentation is an inherent part of the data aquisition and printing process. We are simply mostly ignorant of what is happening.
For instance, on a PC, if I bring up the *same* image in Micrografx Picture Publisher, Adobe Photoshop, or Microsoft PowerPoint it can appear *different* on the monitor. Why? Because Picture Publisher incorporates gamma correction for a monitor while PowerPoint may not. Thus, if I print the image using PowerPoint, I explicitly have to manipulate the gamma when I set up the printer. If I print the same image from Picture Publisher, I don't, because calibration is something you can do as part of a default setup. Now, does that mean that I have "modified" the image if I print from PowerPoint, but did not "modify" the image if I print from Picture Publisher? If I calibrate my printer before I print an image, does that count as "manipulation?" If I calibrate my printer using a different calibration sheet does *that* count as manipulation? If I use a display that automatically does a histogram stretch to fit the display device does that not count as "manipulation" since it's written into the software or firmware, but if I use another display setup and have to hit an "optimize display" button it suddenly becomes manipulation?
Personally, I don't think that any of these sorts of basic optimization exercises should be "counted" as "manipulation," any more than the calibaration and optimization of any device is considered "manipulation" of the data. Indeed, it's just the opposite -- one should be criticized for not doing appropriate calibration and optimization. If one includes all such "manipulation" one might as well publish the technical manuals of all the devices used in the image pipeline. If one defines anything done by hand as manipulation and anything done in ignorance as not manipulation, the idea of manipulation has no meaning.
Thanks to everyone who has replied about my query. I'm passing these answers on to the researcher involved and will let her pursue it further. Obviously, the question needs to be narrowed down considerably, so I'll let her take it from here. Many thanks again.
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
We have an ITI MFG board with the AM-DIG input module (and Xillix camera) and we are looking for software that will, at the very least, permit us to control and grab frames with the camera. We own IPProWin(v1), Snapshot Plus and Mocha but these lack the appropriate drivers and the manufacturers consider both the hw and sw obsolete and unsupported.
Is there anyone who can supply those drivers and/or other sw?
Most of my work is for use within the company so the objectives for presenting images lie not so much in how the image has changed from the original, but rather, does the image clearly describe the issue under scrutiny. For most of my customers, altering an image is acceptable if doing so enhances the story that picture tells. However, most of my image adjustment is restricted to changing the image for printing or presentation purposes, i.e. brightness, contrast, etc.
There are cases where I perform no manipulation at all: patent support and legal support. I believe it is my ethical responsibility to retain the integrity of the data. Taking this attitude ensures that my integrity is retained. So far it has worked.
By the way, take a look at the cover of the most recent issue (Mar. 12, I believe) of the well-respected, peer-reviewed scientific journal, Nature. It contains a colorized electron micrograph of a microcircuit. I think it is ethical, descriptive and attractive.
Harold J. Crossman Senior Scientist OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
} } ...... Like you, I don't think we ought to be } including standard color charts with our presentations, but in many cases we } should be prepared to. At the risk of being defensive, my suggestion of } putting some type of unabtrusive "histogram manipulation strip" with a } processed image was along the lines of putting a micron bar with a SEM } micrograph ... sometimes there is no need, sometimes there is a ^very^ real } need. } } cheerios, shAf }
No reason to be defensive. I guess I came off a little strong. I suppose that's because a good part of what I do is in forensics. In that community, there seems to be this grasping at straws for a "real" image that, simply, doesn't exist. All images are simply collections of artifacts, and different methods of acquiring and manipulating images (digitally or chemically) simply provide different relationships between those artifacts.
The *real* question is whether or not the image accurately shows the information and/or relationship you want to show.
Thus, for your example, a "histogram manipulation bar" would be perfectly appropriate for an image comparison in which how the manipulations were done would strongly affect the results being reported -- say, comparing the grey level in picture A with the grey level in picture B. Such a bar, however, would not be needed in a more general illustration, such as one that shows simply the presence or absence of a feature.
As an example, in traditional microscopy, a person doing densitometry does need to show that there has been correction for nonisotropic illumination, black level noise, etc. However, a person who is putting a picture of a viral inclusion in an article simply to show that he or she saw a viral inclusion need not obsess about whether or not the color balance is *exactly* like that he or she saw through the scope -- and moving from color to black and white is an unavoidable "manipulation" which involves a bizillion potiential decisions. It simply isn't important. Even more, if the author does a histogram stretch to make the illustration more informative, more power to him or her. In that case such a bar is a distraction.
To all manufacturers of microscopes and related imaging equipment/software:
"What's new at M&M '98?"
If your company will be exhibiting at Microscopy & Microanalysis '98 *and* has new technology to present, please send us the information! American Lab has given us space for a special "Focus on Microscopy" article in their July issue (circulation: 150,000).
We need:
1. Company name, contact, phone, address
2. a 25-50 word description + picture (optional)
3. M&M Booth number
Deadline for receipt of materials: April 3
Space is limited and will be filled first-come/first served
Copies of last year's are available on request from our office. This year's article will focus just on new technologies.
Disclaimer: This is not a commercial message ... MME receives no compensation for this article.
I plan on purchasing a video printer to be used on a JEOL 1200EX TEM and a Topcon DS130 SEM. I would very much appreciate any advice on which model to buy or not to buy. If anyone has any recommendations, please e-mail me at acarol1-at-uic.edu .
I'm one of thoughs people who grew up with silver based photography, then in the career of scientific imaging has had to switch to digital photography. I think it is interesting that the digital revolution has brought out the question of ethical manipulation, since traditional photography has a long tradition of manipulation and has dealt with this problem for nearly 160 years. The issue is the same for both imaging systems: Enhancement vs. Falsification.
Enhancement is for the sake of clarity. Falsification is selectively changing the information.
It has been acceptable to increase contrast for the sake of clarity, however it is unethical to selectively inrease the contrast in just one region of the image.
The digital image just makes it easier to selectively manipulate the image, but it has all been done before in the traditional photography it was just more difficult and still as unethical as it is now for digital images.
The film response to light was grossly manipulative yet people accepted it as a real representation. A person may have photographed a control slide for comparison to the experimental fluorescent slide by just exposing them for the same time on the film. However, due to reciprocity failure of the film this may be a gross falsefication of the data.
So the bottom line is still the same: We still need to be held to a standard of ethics, and that standard is getting stronger not weaker. The results of our labors still need to be repeatable by another person and held to a critique.
For me just the fact that response to light can be linear is fantastic!!
Bob Derm Imaging Center
On Thu, 19 Mar 1998, Humphrey, Charles wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Other Microscopists, } } The issue of ethical digital image manipulation was discussed at } previous MSA-MAS meetings but I still have questions. } } What is appropriate manipulation of microscopy images (particularly } grey-scale TEM and SEM) for scientific and public relations purposes? } How acceptable is pseudo-colorization even for public relations? Are } painting programs as appropriate for this purpose as the scientific } packages that use grey-scale to color manipulation for the whole image? } Is discretionary painting of parts of the image as acceptable as the } placement of an arrow to highlight specific parts of an image? For } public relations purposes with non-microscopist scientists and others, } how much information is needed to explain the digital manipulations } used? } } There are other questions that may be asked but this is a start for } discussion. } } Curious and concerned about ethical limits, } } Charles Humphrey } CDC } cdh1-at-cdc.gov }
I cuncur with the previous posts... That manipulation to enhance appearance without changing meaning is a moot problem. If the meaning of the image changes, then disclose the manipulations.
This is not really new. Digital imaging makes changes easier, but who has never used a filter on a film camera. Was that manipulating the image? Are tungsten light filters unethical :)
Woody White McDermott Technology, Inc woody.n.white-at-mcdermott.com http://www.mtiresearch.com
Home woody.white-at-worldnet.att.net http://www/geocities.com/capecanaveral/3722
} By the way, take a look at the cover of the most recent issue (Mar. 12, } I believe) of the well-respected, peer-reviewed scientific journal, } Nature. It contains a colorized electron micrograph of a microcircuit. } I think it is ethical, descriptive and attractive.
I have published several biology/morphology papers in which the images were derived from digital cameras and, of necessity, heavily processed. In some cases, the images were derived/sythesized from digitizer tablet input. In all cases, they were completely documented as to the underlying procedures.
After all, I used to do my manipulations in the darkroom......
} } On Thu, 19 Mar 1998, michael shaffer wrote: } } } } } } What is appropriate is relative to (... in my mind ...) appropriately } } } presenting the image's history of manipulation. ... } } } } } } } } } But that really begs the question, or rather, ignores the question. *Any* } } image that is digitally acquired or printed is "manipulated." That's what } } color management *is,* for instance. } } } } [skip] } } } } Your points should be well taken ... 'cept issues like "color } management" and "calibration" should also acknowledge what we've learned to } accept and trust, as long as we have some faith in the imaging researcher.
If one trusts that the same final image can always be obtained from a raw image file (which should still be accessable somewhere--just like original notebook data), that is equivalent to trusting the resear- cher. That is, if I were able to see the raw image and the researcher told me the steps to follow, I could obtain the final image myself. I might never want to do it, but as an editor or reviewer, I'd like the option of reproducing the final image for myself to see what each mani- pulation did to the clarity, etc.--especially if the image purported to prove a controversial point.
} Such manipulations have long been a part of the image presentation process } ... "which film developer did you use?" ... "what contrast paper did you } use" ... "How accurate did your publisher reproduce your plates?".
A photography magazine covered this topic and pointed out all the ways that darkroom images can be manipulated without the use of a computer. Again, it's a matter of trust or having the ability do reproduce the mani- pulations oneself. Of course, the unscrupulous could use altered images to produce fraudulent data, but the usual safeguards of science should (eventually) weed this out.
} The } original question I believe begged the question "How do we learn to trust } digital image manipulation?". Like you, I don't think we ought to be } including standard color charts with our presentations, but in many cases we } should be prepared to.
In any event, every image from initial to final should be archived.
} At the risk of being defensive, my suggestion of } putting some type of unabtrusive "histogram manipulation strip" with a } processed image was along the lines of putting a micron bar with a SEM } micrograph ... sometimes there is no need, sometimes there is a ^very^ real } need. } Perhaps the histogram can also serve as a micron bar :-). Yours, Bill Tivol
A few days ago someone posted about a KEVEX EDS system from a JOEL840 SEM they were looking to get rid of. Unfortunately I forwarded the email message onto someone who is interested but his harddrive crashed and he lost the message. If the system is still available will you let me know?
Thanks
Tom
Thomas Mullarkey Murray email:tm8a-at-virginia.edu Thornton Hall - MSE phone:(804)982-5659 University of Virginia Fax: (804)982-5660 Charlottesville, VA 22903
I am a fringe browser of Microscopy, using basic light microscopy in an applied (and basic) manner - and have a general inquiry about Image Analysis Systems (microscopes, CCD cameras, and software).
I am deciding on purchasing a system to be based here in Auckland. While checking out local systems, I am also interested in hearing anyone's views or preferences as to the components/manufacturers. I have lists of microscope and camera manufacturers, but would appreciate comments from your experience with the use, servicing, etc, of such equipment (and recommended software).
The system is to be used primarily for (semi-) quantitative analysis of tissue sections stained in a variety of ways. Current labels are not fluorescent, but I can see this being useful in the future. The size of things we will actually be measuring ranges from a few microns (about 10) in length, and up to linear distances of a few hundred microns, or a few thousand square microns. Final magnification ranges from about 150X to not more than about 2000x. We will need digital images of adequate resolution for electronic transfer for publication, and can find a suitable printing device when hardcopies are necessary.
If you can be of any help, please send your reply directly to me. I can post a summary to the list if anyone expresses interest in the responses.
Thanks in advance for your time.
Heather
Heather K. Smith, Ph. D. Dept. of Sport and Exercise Science University of Auckland Private Bag 92019 Auckland, New Zealand
I would like to improve my double-labelling techniques for vesicles and cytoskeletal elements in cultured PC-12 cells for fluorescence microscopy. For vesicles I have been using monoclonal anti-synaptophysin conjugated with FITC, and for microfilaments, I have been using rhodamine-phalloidin.
I have been having some difficulty with the "fixation followed by permeabilization schedule"-vesicles and the CSK both stain, however...vesicles appear to clump as if they are leaving the cytoskeletal mesh as found in the cortical peripheries. We believe that the irregularly shaped tubulovesicular structures are Golgi or ER, so that part seems OK. The situation at the cells' cortices make it appear as if there is apocrine secretion, but my cells are indeed merocrine. We want to see where the vesicles are localized with respect to their exocytosis at different time frames with agonist stimulation.
Does anyone out there have a good protocol for this? I have been using a schedule of a modified Karnovsky's fix in sodium cacodylate (worked for LM well before for CSK labelling exclusively) for an hour followed by a PEG 8000 wash and a 1.25%/0.2% Triton-X 100 detergent (in PHEM buffer) permeabilization for 10 minutes. Shall we try a phosphate- buffered saline instead of the cacodylate, which is routinely used for EM? Osmolarity problems? Too long of permeabilization time?
Any suggestions are helpful-thanks in advance for your help.
Cheers, Vickie
-- Vickie A. Kimler, Ph.D. Assistant Professor of Biology and Allied Health Director, Cancer Research Facility Mercyhurst College 501 East 38th St. Erie, PA 16546 Voice: 814-824-2169 FAX: 814-824-2188 e-mail: {vkimler-at-mercyhurst.edu}
I have had many conversations regarding digital imaging with a wide variety of end users. The points made here have been validated in those conversations. I thought that the point of view from law enforcement might be of interest.
When a photographer or law enforcement agent testifies as to what is seen in a given image, they are attesting to the fact that the image is an accurate _representation_ of what they saw and photographed at a given point in time at a given location. Based on the perceived reliability of the witness, a judge or jury determines whether that testimony is accurate.
There has been much discussion in Law Enforcement circles about digital watermarks and the like that would 'disappear' if an image were manipulated. One well known digital camera manufacturer who indicated that they had a proprietary file format that could not be manipulated was corrected by a hacker. Given the right equipment and talent, an analog film image could be scanned in, manipulated and output back to film and the best experts in imaging could not be able to tell and they will admit that. Someday, it may be possible, but not today. So, my point is that any image, digital or analog, is only as factual or accurate as the integrity of the person representing it as such.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation "See What Develops" 4525 Leonard Parkway Richmond, Virginia 23221-1809 804 254 1011 804 254 1013 Fax warrenj1-at-polaroid.com
In response to Robert Underwood's posting, I can only agree wholeheartedly. As a silver-based photographer over the past 30 years, I have seen these debates repeated many times. As a former journalistic photographer, I can say that image manipulation in terms of contrast, brightness, color, exposure, point of view, timing, etc., etc., etc. is accepted as long as it does not alter the meaning of the reported event. More than this enters into the realm of "art", and I wouldn't touch that question with the proverbial ten foot pole. Even this is an almost impossible ideal, since the very act of pointing a camera in one direction and not another, at one angle instead of another, using one lens instead of another, taking a picture at one instant instead of another, can significantly alter the viewer's perception of the photograph. There is a large literature on this subject, dating back to the origins of the technology.
The problem in terms of scientific "objectivity" (if such a thing exists---I have my strong doubts) is similar, but perhaps even more important.
A rule among journalist photographers is to clearly state any manipulation of the image that would affect the viewer's interpretation of it. Generally, this is confined to such things as directing the action in a photograph (as opposed to simply recording events without interfering in them), or such obvious things as combining separate photographs into one image, and so on.
I would suggest reviewing the literature on ethics in photojournalism and documentary photography, as a starting point.
Relating all of this to microscopy, it is just as clear here as it is in other types of photography that "objective images" are unattainable goals. Specimen selection, fixation protocols, choice of image fields, and so on, are all subjective decisions, varying from researcher to researcher, and this is before we even get into the photographic or digital imaging process. The variables are myriad, much as we would wish otherwise. The responsible researcher tries to control them as much as possible, but we are all cursed with our individualism. We are dealing with a question which is as much in the realm of philosophy as science, and that makes many scientists extremely uncomfortable.
My suggestion: we should continue to record our research as faithfully as we know how. If we are trying not to deceive, we probably won't. Don't use Photoshop to delete unwanted blots on gels, don't try to eliminate uncomfortable peaks on EDS spectra, don't add information that was not there in the actual results, just because we "knew" it should be there, and we should be okay. Those who do try to deceive should hopefully be caught by the tried and true process of replicating their results by more ethical people. I don't see any other way.
Randy Tindall
In responsible photojournalism Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
We have a JEOL JXA8600 microprobe with four WD and one ED X-ray detectors. The 'scope is controlled by an old Tracor computer system and we are concerned that it may die. Presently we've only found Noran (who could supply a Sun-based replacement datasystem) but we'd like to make a choice over the possible replacement, preferably including a PC-based alternative.
If anyone out there is aware of any systems able to support this type of kit, please contact me!
Simon
---------------------------------------------------------------------- Dr Simon Dumbill, AEA Technology plc, 220, Harwell Didcot Oxfordshire OX11 0AB tel +44 (0) 1235 434245 UK fax +44 (0) 1235 435941
} I am sure that the cellulose is a lot softer than the resin. From TEM } or SEM the resin has penetrated through the sample, where it could } penetrate. I am using a diamond knife cutting from 0.05 to 3 microns. } The same problem seems to exist at all thicknesses. The real problem } that I think I am having is that after I float the sample - as the } sample dries it shrinks or expands then ripping the sample apart. } } I have heard of the LR white before. Do you know its components and } properties (e.g. what resins is it made from, shrinkage factor during } curing and the hardness...) } } Thanks for your help and the help to come. } } Sincerely, } Robert Dickson
Robert -
LR White is a proprietary resin, so all that I know is that it's an acrylic. Shrinkage? Low, but a bit more than epoxies. Acrylics are mostly linear polymers without the crosslinking of epoxies, so they tend to section better in your "thick" range. If you do EM, a support film is essential, since acrylics have less beam resistance. LR White is as liquid as water, and is very water-tolerant, so it will infiltrate the hygroscopic cellulose fibers (give it some time and a couple of changes, tho). The "hard" grade is similar to a "medium" epoxy; the basic rule is still to try to match the resin hardness to the sample. My experience at U.C.Berkeley & with the RMC Materials Microtomy course is that it solves lots of comparable embedding problems.
I asked about the knife that you use because you may have another problem. If you've used that diamond for several attempts at sectioning the poorly epoxy-embedded fibers, that means that you've probably smeared an invisible film of uncured epoxy on the knife edge, since unreplaced water in the fibers will interfere with epoxy polymerization. That sticky film must be removed or you will continue to get excessive wrinkling and compression of ALL sections, even from good blocks.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
To the List: clink, clink... my two cents worth....
One should start at the beginning concerning image manipulation.
The histological image as seen in the microscope bears little resemblance to life. It is dead, fixed, chemically altered, dehydrated, stained, sectioned, etc. Why concern yourself with darkroom filters? How long was the tissue in the stain, at what concentration, how often had the stain been used, what temperature was the stain used; how thick were the sections, (gee, what one can do with resolution here), and then of course, what filter sets were being used in the microscope, what kind of coatings were on the optics, what kind of optics in the microscope; what was the age and lot of the film used. The tissue has been manipulated a considerable amount before it ever gets to the darkroom or to the computer.
What really counts is the ethical nature of the investigator to report the results honestly and accurately. And for reviewers to do their job earnestly and honestly.
Blystone in Texas
Robert V. Blystone, Ph.D. {RBLYSTON-at-Trinity.edu} Professor of Biology Trinity University San Antonio, Texas 78212 210.736-7243 210.736-7229 FAX
My boss returned from a conference raving about a display he had seen of ESEM images of pharmaceutical solids. The question naturally arose as to why my puny efforts seemed to fall far so short. I spent a day or two considering alternative ways of supporting my family before we learned that the images were heavy on the digital cosmetics. I assured the boss that henceforth all is well! Photoshop to the rescue!
I think any image is manipulative to some degree. But then so is microscopy. Why did we chose that particular field of view over the hundreds or thousands we may have seen on the scope? Evidently, because it illustrated some point we wished to make. All this is in the nature of microscopy. What is important, in my mind, is that the methods of making the image are documented appropriately and available to others. I do not object at all to the other companies display - it was informative and instructive as well as aesthetically pleasing. I was, however, glad that they were willing to share some of the methods that went into making the images.
Thanks Robert A. Carlton Robert.Carlton-at-RP-Rorer.com Tel. 610-454-3949 Fax 610-454-5990
I have an immediate need for a plot of instrumental resolution/capability for e.m./etc(SPM) instruments as a function of the analytical needs in the semiconductor processing world--past, present and future projection.
It's a case of where I can swear I've seen such plots in a thousand places but now that I need one I can't find one.
If you can help, please send it to me in any digital format as an attachement to my Lotus Notes address anderron-at-us.ibm.com Please grant permission for me to use it and I will give full credit, etc. It is heading to my invited talk in the "Microscopy of Semiconducting Materials" session in the M&M98 meeting.
As promised, here is a summary of replies to date to my queries about the Iris Explorer.
I asked these questions because, like many of you who responded, I was interested in finding out whether the Iris Explorer would be suitable for our work. I had never heard of it before, and we have been looking at image analysis systems for a long time. In previous extensive discussions on this Bulletin Board, no one had mentioned it.
A couple of very helpful colleagues have pointed out that, just as we suspected, the Iris Explorer is actually used to display computed data. Although it has image analysis capabilities, it is not meant to be image analysis software. It can be useful though, if you want to turn numerical data into images, as some of the people who responded did.
If you are interested in getting more information, here is the website you should visit - it also has tutorials to try:
http://www.scs.leeds.ac.uk/iecoe
Thanks to everyone who replied about this. Thanks too, for all those who gave their comments on different image analysis software - we are still looking...
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
} } The } } original question I believe begged the question "How do we learn to trust } } digital image manipulation?". Like you, I don't think we ought to be } } including standard color charts with our presentations, but in many cases we } } should be prepared to. } } In any event, every image from initial to final should be archived. }
I'll even take issue with this claim. Consider the following two scenarios:
First scenario:
I have an interactive image-processing tool with a nice GUI which allows me to see the effects of various algorithms quickly. Let's say I am doing an adaptive histogram equalization and I have control over the size of the adaptive regions in each dimension (e.g. the regions need not be square but could be rectangles), the shape of the adaptive regions (they could be rectangles or circles), and the degree of contrast clamping (a limitation on the contrast enhancement based on the statistics of adjoining regions). So, I have four parameters, each of which has a large number of possible values.
So, I sit there and twiddle with the parameters until I get what I consider an visually optimal image. Let's say that I do a hundred twiddles. Does this mean that I should save every one of those rejected manipulations since they were in the event line between the original and the final? Of course not.
Now, let's say that I *don't* have a nice GUI, but have to run it from the command line. So, instead of twiddling dials a hundred times, I run the program a hundred different times, generate a hundred images, and choose the one that looks best. Should I archive all hundred of these images? Well, if it is silly to generate an archival image for every twiddle of a GUI widget, it is equally silly to save all the results of all the runs of all the manipulations one does.
Second scenario:
Let's say that I have a nice GUI that incorporates all three major variants of adaptive histogram equalization. One of these variants is called "sharpened histogram equalization" by some, in which an unsharp mask is performed either just before or as part of the equalization. Much like adaptive histogram equalization (AHE), unsharp masking is highly parameterized.
Even more, however, unsharp masking is a rather separate kind of process, even though it can be combined with equalization. My tool, however, puts them in the same pipeline. So, now I am twiddling unsharp masking and AHE parameters in my little GUI. Once again, am I obligated to save an image for archive every time I turn a dial? Once again, of course not.
Now, let's say, again that I don't have a fancy GUI, but run both from the command line. I first run an unsharp masking program, and then take the result of the unsharp masking program and run it through an AHE program. Let's say I run the unsharp masking 20 times and the AHE 20 times and generate 400 images, of which I choose one. Am I obligated to save the unsharp masking runs as separate archived images because it is a "different" process?
I don't think so. Surely, the presence or absence of a fancy GUI should not be the determinant of what constitutes an intermediate image and what should be archived.
Even worse, consider what happens when you use development environments which use large intermediate data structures. Is one obligated to archive all those intermediate data dumps? For instance, I recently processed a 2Kx2K 3-color image in which I had the following pipeline in AVS (Advanced Visual Sytstems -- a development environment):
read_in_image (a module or small program) | | (these little bars denote taking the data from one | program and taking it to another) | | change from byte to float data (1 more module) | | | V transform image from (1 more module) rgb to Lab color spaces | | | V separate color channels (1 more module) | | | | | | | | | V V V make wavelet stack (three instantiations -- one for each channel) | | | (approximately 20 more modules) | | | | | | | | | V V V separate portions of each wavelet stack (say three parts) | | | | | | | | (nine instantiations -- three for each | | | | | | | | channels) (approximately 12 more modules) | | | | | | | | V V V V V V V V process each portion with a separate image processing algorithm -- with twiddling of parameters (approximately 30 more modules) | | | | | | | | | | | | | | | | | | | | | | | | V V V V V V V V construct wavelet packets (approximately 30 more modules) | | | | | | | | | | | | | | | | | | | | | | | | V V V V V V V V do inverse wavelet transform (three modules) | | | | | | | | | V V V combine color channels (1 module) | | V transform from Lab to rgb (1 module) | | V transform from float to byte data | V write image (1 module)
Now, because of the development environment I used, I didn't have to write any intermediate images. However, I write my software so I don't have to rely on the development environment, and I could have *also* written this pipeline as a UNIX script and written and read intermediate images to and from disk.
Even though I have *no* intermediate images that are written to disk, I have little image viewers and a GUI that allows me to control the data flow, see intermediate results, and twiddle parameters all along the way. All the intermediate data is held in RAM and virtual memory. So, in this pipeline, what constitutes an intermediate image that "must" be archived? Would it be different if I ran it using the command line pipeline which explicitly writes out the intermediate images?
Personally, I think the idea of having to save every intermediate image is not merely a bad idea, it is unworkable. It means, in large tools, arbitrarily writing out "intermediate" images which have no use. Even worse, writing out such intermedate results may make it impossible to do the work. Were I to write out an intermediate image after each one of these steps, even using the UNIX command line method, I would use up over 7 Gigs of disk space per run per image per "twiddle". When I run this as a UNIX pipeline, I delete each intermediate image as soon as it is read by the next program in line.
Certainly, one should describe what one has done, but to me, the above diagram fleshed out to include the actual processing algorithms and the *final* parameterization is enough.
I don't archive all these "intermediate" images. I archive an adequate ascii description of the pipeline and the final results.
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A San Joaquin Delta College Faculty member of the Microscopy Technology = Center is planning a Sabbatical Leave for the Fall '98 semester. The coll= ege seeks an instructor(s) to cover her courses. MINIMUM QUALIFICATIONS: Bachelor's Degree plus two years of directly related experience OR an = Associate Degree plus six years of directly related experience OR a valid = credential. DESIRABLE QUALIFICATIONS: Master's or PhD Degree in a Biological = Science; Experience in teaching Electron Microscopy COURSES TO BE TAUGHT: Introductory Techniques for Transmission Electron Microscopy (EM21) This is a lecture/lab course which includes beginning Transmission Electro= n Microscopy dealing with the alignment and operation of the TEM, vacuum = techniques, photographic techniques, as well as the preparation of = particles and replicas for viewing in the TEM. Includes individual = training in the use of the TEM, preparation techniques, and written and = oral reports. (Lec - 2 hrs; Lab - 3 hrs/wk) Biological Ultrastructure (EM28) Course contents include specific information about the fine structure and = function of cells and tissue at the ultrastructure level. Videos, slides = and micrograph examination will be correlated with the lectures so that = students will learn to recognize the fine structure of cells and tissues = in relationship to their function. (Lec-2 hrs/wk) Current Microscopies: Optics, Theory and Application (EM30) Course contents include information related to the physical laws and = applications of the various types of current microscopies e.g. = TEM,SEM,FIB, AFM, and confocal microscopy, as well as other current = topics e.g. asbestos analysis, lab design, etc. (Lec - 2 hrs; Lab - 3 = hrs/wk) Advanced Techniques in Biological Electron Microscopy (EM37) Course contents include lecture and laboratory which covers advanced = techniques for biological specimen preparation in TEM including an = advanced research project.( Lec - 1 hr; Lab - 6 hr/wks) TERMS OF EMPLOYMENT: Adjunct / Non-tenured track position. APPLICATION: Contact Human Resources at 209/954-5056. DEADLINE: The Screening Committee will begin to review applications on = April 22, 1998
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A friend asked to post a question concerning picric acid disposal. Is it possible to easily convert it to a non-explosive form by reacting it with something in the lab. I suggested mixing it with protein. The problem is that it is radioactive as well as explosive so th safety people don't know how to handle it. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
After I posted the summary, our Statistician posed the same question to the distributors of the Iris Explorer.
Patrick Craig, from the Visualization group of NAG, Ltd. replied and said that it is possible for the Iris Explorer to handle images in several formats including TIFF and GIF.
I guess the bottom line is that there are alot of easy to use IA software packages out there which would be able to do what is required. The Iris Explorer is a very sophisticated tool which is also very powerful in a number of different applications.
Patrick Craig can be reached at : infodesk-at-nag.com
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Argonne National Laboratory currently has an opening for a materials scientist at its site in Idaho Falls, Idaho. The main responsibility of the position is characterization of ceramic and metallic waste forms produced in an electrometallurgical spent nuclear fuel treatment process using transmission electron microscopy. An additional responsibility is maintenance (to the extent not covered by a service contract) and upkeep of a JEOL 2010 transmission electron microscope and associated sample preparation equipment. This position will involve work with radiological materials. A Ph.D. degree in materials science, materials engineering, or ceramic engineering with extensive experience in transmission electron microscopy of ceramic materials is essential. The successful candidate will posses strong written and oral communications skills and a proven ability to collaborate with others. U.S. citizenship and ability to obtain a U.S. Department of Energy security clearance strongly preferred. Argonne National Laboratory is an equal opportunity employer.
Interested applicants please send cover letter and resume to:
Dr. Terry Totemeier Argonne National Laboratory P.O. Box 2528 Idaho Falls, ID 83403-2528 terry.totemeier-at-anl.gov
Hello microscopists, Does anyone know where I can purchase a cheap, used microtome (not an ultra microtome) for cutting 100 micron sections of rubber, or plastic material? I was hoping for something in the 200 dollar range.
I would like to improve my double-labelling techniques for vesicles and cytoskeletal elements in cultured PC-12 cells for fluorescence microscopy. For vesicles I have been using monoclonal anti-synaptophysin conjugated with FITC, and for microfilaments, I have been using rhodamine-phalloidin.
I have been having some difficulty with the "fixation followed by permeabilization schedule"-vesicles and the CSK both stain, however...vesicles appear to clump as if they are leaving the cytoskeletal mesh as found in the cortical peripheries. We believe that the irregularly shaped tubulovesicular structures are Golgi or ER, so that part seems OK. The situation at the cells' cortices make it appear as if there is apocrine secretion, but my cells are indeed merocrine. We want to see where the vesicles are localized with respect to their exocytosis at different time frames with agonist stimulation.
Does anyone out there have a good protocol for this? I have been using a schedule of a modified Karnovsky's fix in sodium cacodylate (worked for LM well before for CSK labelling exclusively) for an hour followed by a PEG 8000 wash and a 1.25%/0.2% Triton-X 100 detergent (in PHEM buffer) permeabilization for 10 minutes. Shall we try a phosphate- buffered saline instead of the cacodylate, which is routinely used for EM? Osmolarity problems? Too long of permeabilization time?
Any suggestions are helpful-thanks in advance for your help.
Cheers, Vickie
-- Vickie A. Kimler, Ph.D. Assistant Professor of Biology and Allied Health Director, Cancer Research Facility Mercyhurst College 501 East 38th St. Erie, PA 16546 Voice: 814-824-2169 FAX: 814-824-2188 e-mail: {vkimler-at-mercyhurst.edu}
If you are interested in immunocytochemistry, immunohistochemistry or other affinity labelling, then check out sci.bio.immunocytochem
sci.bio.immunocytochem is a "Usenet newsgroup" and therefore you need to have access to Usenet in the first place! Check with your academic computer department, or your commercial service provider if you have access to Usenet. Most good Internet accounts include Usenet access.
You can then set up your Internet browser to read and post messages to any newsgroup, including sci.bio.immunocytochem; or you can get some freebie newsreading software.
"Subscribing" to a newsgroup merely changes its status in your newsreader, and makes it easier to find your favourite groups amongst the thousands of newsgroups available. You dont have to subscribe to sci.bio.immunocytochem in order to read or post messages.
You just get your newsreading software to "get new messages" in your chosen newsgroups whenever you want to read the latest messages; there is no e-mail involved, unlike mailing lists!
You can post a new message, or a "follow-up" message to sci.bio.immunocytochem
E-mail me if you have any hassles with Usenet access etc!
We are trying to locate, for purchase, a used epi-fluorescence attachment, with HMX lamphouse,lampsocket, for mercury HBO 100 watt bulb, also B-G filter. This is to fit onto a Nikon Diaphot microscope, 1985 vintage.
If anyone has one that they are willing to part with please contact me offline.
Thank-you for your help.
Pat Glazebrook MetroHealth Medical Center Rammelkamp Center for Research and Education email: pglazebr-at-research.mhmc.org phone216-778-8958
FEI Company has the following position open to be based in Austin, Texas:
Job Title: Field Service Applications Engineer Geographic Location: Austin,TX
1) Job Summary: Conduct on-site and in-house operational training for new and existing users on the full range of FEI's systems. 2) Essential Responsibilities: A) Conduct on-site operations training for new users of FEI's systems for new installations and for existing installed base as required. B) Give technical assistance to worldwide customers, engineers, and/or representative for the full range of products. C) Conduct fundamental training courses in focused ion beam and scanning electron microscope applications for filed service engineers and technical support specialists. D) Define on-site customer training programs for the full range of FEI systems going into the IC market. E) Submit paperwork accurately and in a timely manner. 3) Secondary Responsibilities: F) Advise and recommend actions to the Quality Improvement team and/or development team on any operational issues seen in the field. G) Assist with customer site acceptance. H) Other duties as temporarily assigned by department manager. 4) Specific Job Skills: I) Strong customer service focus. J) Self-starter with exceptional attention to detail and follow-up. K) Superior written, verbal and interpersonal communication skills. Able to converse readily in English. Other languages a plus. L) Solid organizational abilities. M) Desired scientific or engineering background, including familiarity with ISO900 standards. N) Training background preferred. O) Eligible for passport. P) Must have a valid driver's license. Q) Able and willing to travel up to 75% of the time. 5) Minimum Qualifications: R) BS or equivalent in Physical Science; MS preferred. Physics a plus. S) 3-5 years experience in focused ion beam or scanning electron microscope operation. T) Proven ability to work cooperatively in an interdisciplinary team environment. U) Understanding of DOS and Windows operating systems as well as functional knowledge of SW programs such as Excel, Access and MS Word. V) IC industry background strongly preferred.
Join FEI Company and benefit from out compensation and benefits packages which include fully paid medical, dental, vision, and life for employees and their families, profit sharing, tuition assistance, employee stock purchase plan, wellness program and 401(k) with match. Please submit resume to Lisa Olivia either by FAX: 503.640.7509 or email: lolivia-at-feico.com.
Lisa Olivia Recruiter FEI Company PH) 503.844.2601 FAX) 503.640.7509 email: lolivia-at-feico.com
I hope I am not breaking any rules about sending a JPG image to the list server, and further hope that only one of you will let me know if I am. I have attached an image of a rotary shadowed molecule present in large quantity in a collagenase digest of mouse skin. It seems to me that I have seen a similar image some time ago in the literature, but I can not place it. Does anyone know what molecule I have imaged?
Thank you in advance!
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
Patricia A. Glazebrook wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi- } } We are trying to locate, for purchase, a used epi-fluorescence attachment, } with HMX lamphouse,lampsocket, for mercury HBO 100 watt bulb, also B-G } filter. This is to fit onto a Nikon Diaphot microscope, 1985 vintage. } } If anyone has one that they are willing to part with please contact me } offline. }
Pat,
I am curious... "used" for price reasons or because of availability? While the original #78947 is long gone; several third party manufacturers have machined a replacement and the rest of the parts (cubes, lamps, power supplies, etc.) are all still current. If you have not done so, please contact your local Nikon Representative/Dealer in Ohio (I think area code 216 is Ohio) and that would be the Fryer Company (513)851-2992. They should be able to help you find this part.
I find the discussion about scientific image manipulation rather interesting, but, here's a thaught:
Please correct me if I'm wrong, but I seem to remember that, at least som= e time ago, all scientific findings and research results, in order to be considered as valid, should be reproducible by another researcher. If that's not the case (and just think of all those who claimed to have achieved cold fusion, but other people, or even themselves, were unable t= o repeat the experiments), the findings or results would just not be taken = as valid. Doesn't the same scheme apply to whatever results that are based on or supported by image contents?
Microscopists, I was wondering if anyone could tell me either the best way to prepare and mount bacterial for use in an SEM or a location where I could look to find such a prep.
SALZBURG, 22nd of March, 1998, local time 01.20 p.m.
Sir, dear Greg, may I cite from an MSDS (MERCK DARMSTADT, FRG):
PICRIC ACID: 2,4,6-Trinitrophenol, C(6)H(3)N(3)O(7), M=3D 229.11 g/mol; melting point: 122 degr. C; solubility in water limited; ignition point: = 300 degr.C;=20 thermic decomposition(and therefore } } explosive { {): 300 degr.C and up = (especially if you heat the compound very rapidly); } } toxic { {; BUT: I am not aware about any } } radioactivity { { of the substance (why picric = acid should be radioactive?? unless you use isotopic forms of C??).
hazardous decomposition products: CO (carbon monoxide), = nitrose(o)-gasses; hazardous reactions: with alkali-hydroxides, alkali-metals, metals, = salts of metals, fluor.sensitive to shock; inappropriate materials to = use: metals.
Dry picric acid crystals are more dangerous (with respect to be an = explosive hazard) than wet crystals: therefore:=20 Original packing: picric acid crystals come with 0.5 ml H(2)O/g solid=20
Safety instructions and safety tips (excerpt): =20 KEEP CONTAINER SCREWED TIGHTLY; AVOID CRUSHING/BLOWING and FRICTION (rubbing). AVOID skin contact: = danger in case of resorption through skin; storage: tightly closed, at = room temperature (+15 to +25 degr.C); Keep the compound wet.
Keep in mind: you normally don't store (should not be storing) big = amounts of picric acid in your Lab. For histological applications (like = BOUIN's fluid) or for TEM-fixatives like FA-GA-PA-mixtures I would = suggest 50 g would be sufficient. In my opinion and experience of over 15 years EM-Lab there would be no = obvious or extreme hazardous situation, if the compound is stored = appropriate as indicated above and the container is/has been tightened = closely every time you used it. And: the amount and how it is stored = (and one works with it) makes a difference in the way of acting as a = dangerous chemical (with respect to explosions or ignition) =20 Disposal: Dilute with water (this would normally be the case, if used = for histology purposes), don't dispose of into the/a water drain system = (picric acid is a water hazard class 2[european classification]); = collect separately and dispose of according to national/local = legislation (authorized waste collectors).
I collect hydrous solutions of the stuff in a separate glass flask (or = plastic bottle) tightly closed with screw cap; if a litre or so has = accumulated, it will be disposed of through a certified waste collector. As I said: I have stored an original amount of 100 g Picric acid = crystals ( to which originally are added !! 50 ml of H2O !! ) from 1984 = on in my Lab; now there are left about 40 g: the substance is wet; = stored under ambient room temperature conditions and I don't look = forward to have any problems in the future with its safe storage and = use. The point you are considering for a "safe deactivation" of the substance = by mixing it with proteins IMO seems to be tricky, unless one knows = about the amount of protein you have to add to bind the compound totally = to the protein(s) available.
Best wishes for "your solution" of the problem, Wolfgang
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
A friend asked to post a question concerning picric acid disposal. Is = it possible to easily convert it to a non-explosive form by reacting it = with something in the lab. I suggested mixing it with protein. The problem = is that it is radioactive as well as explosive so the safety people don't = know how to handle it. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 =20 Scientific Director, = =20 ICBR Electron Microscopy Core Lab = =20 PO Box 118525 Fax: 352-846-0251 =20 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ =20 ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
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We find the best way is to use small 0.2um millipore filters. Whatman International do them tel +44 (0)1622 676670 (I have no commercial stake here!)
You can get little ones (13mm) that fit in a special holder that will go on the end of a syringe.
Just squirt your solution of bacteria (probably best washed first in PBS buffer) through the millipore filter (shiny side up) then quickly remove the filter and fix in 3% gluteraldehyde in PBS for an hour or so. wash-pBS osmicate-1% osmium in PBS for 1 hour wash-PBS dehydrate-30%, 70%, 90%, 100%, 100% 20 mins each Then critical point dry your millipore filter (with bugs hopefully still attached) Mount on SEM stub with double sided sticky tape Use conductive paint to make line from surface with bacteria to metal on stub Coat with gold in sputter coater And voila!
Amanda Wilson wrote: ================================================== We find the best way is to use small 0.2um millipore filters. Whatman International do them tel +44 (0)1622 676670 (I have no commercial stake here!)
You can get little ones (13mm) that fit in a special holder that will go on the end of a syringe.
Just squirt your solution of bacteria (probably best washed first in PBS buffer) through the millipore filter (shiny side up) then quickly remove the filter and fix in 3% gluteraldehyde in PBS for an hour or so. wash-pBS osmicate-1% osmium in PBS for 1 hour wash-PBS dehydrate-30%, 70%, 90%, 100%, 100% 20 mins each Then critical point dry your millipore filter (with bugs hopefully still attached) Mount on SEM stub with double sided sticky tape Use conductive paint to make line from surface with bacteria to metal on stub Coat with gold in sputter coater And voila! ================================================== There is a variation on this theme: Use 13 mm membrane filters made out of pure silver instead of the polymer type. The "sticking" ability to the silver is more or less comparable to that on a polymer membrane filter and it can be critical point dried just like a polymer membrane. The advantage is that the substrate is already conductive hence one can always reduce the amount of gold needed in the sputter coater, and in some instances, it can reduced down to zero, with obvious advantages.
If adhesion is in fact a problem, it can usually be enhanced by a short term exposure to oxygen plasma etching (e.g. one or two minutes) to remove adsorbed organics.
Disclaimer: SPI offers the SPI-Pore line of silver membrane filtration prodcuts and and also plasma etchers and they are all described on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
thanks to all who sent info on about testing the vacuum system.
seems i may have more problems:
1) the leak rate from HVac is {3uA/min atm=250uA Hvac=25uA on vacuum meter test points (OK)
2) V1 doesn't work in auto mode, and many times won't work in manual
3) DP heater warms up (light on) in about 20minutes
4) R4 vacuum control pot will not make V1 open when vacuum meter {190uA while in auto mode
5) solenoid valve for the closing side of V1 will not release pressure so that the opening side of the valve (when pressurized) will force the valve to open.... most of the time (related to #2 above)
6) some of the valves (like v4) will actuate manually when the controls are in the auto mode (is this normal...or an indication that the logic is all messed up?)
7) V5 (airlock) will not open in auto mode, but does in manual (as does the associated leak valve)
i'm really confused about this since it seems to be a moving target with many appearent "failures". is there one circumstance or condition that would explain these observations????
thx in advance! brian
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
I am trying to reply to Yvan Lindekens, who had replied to a query about the Lensman. His email address "yvan.lindekens-at-skynet.be" is not accepted by our computers--does anyone have more information?
Thanks, Doug Darnowski
****************************************************************************** Douglas Darnowski Department of Crop Sciences 384 ERML 1201 West Gregory Drive University of Illinois Urbana IL 61801 work ph: (217) 244-6150 fx: (217) 333-4777 home ph: (217) 356-6606 fx: (217) 356-4454 email: darnowsk-at-staff.uiuc.edu
Eric, Here are two ways to prep bacteria for SEM that have worked well for me: 1) Let living sample attach to poly-L-lysine coated slides before fixation. Make sure you spread suspension over the c/s then apply fix directly to it. Check concentration under a phase microscope. Then prep slide as usual for SEM (mark the sample side with diamond scribe!). Usually a Para/Glut combo followed by osmium but check your reference first (I've seen other secondary fix choices, depending on what you are looking for)-remember gentle rinsing. 2) Use a 0.22 or 0.45 um filter (the kind you can separate) Draw live sample through (usually opposite direction of normal filter flow) and then draw fix through and saturate). This is a little more difficult because you can't tell concentration until your on the scope and the filter may fold during CPD. I prefer the first technique.
Good Luck, Michael Delannoy JHMI Microscopy Faclity
Dear all, There has been much debate about how to apply ethics to the particular issue of image processing. I am interested in a slightly more general question, however.
What basis do we use as scientists for deciding what is and is not ethical? It seems to me that this is not a trivial question, especially when we live in pluralistic societies and there are a wide range of different approaches to morality and ethics put forward by different individuals or groups in society.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
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I apologize that the incorrect phone number was listed for Steve D'Angelo = who is selling his MT2. If you are interested, His correct phone number is 650/738-2699 Steve D'Angelo 1107 Galvez Dr Pacifica, CA 94044
MT2 in excellent condition Factory serviced.
He does not have an e mail address.
Please do NOT reply to this e mail address. I am sending this message = for a friend. Thanks.
======================================== Charles W. Allen Electron Microscopy Center-HVEM-Tandem Facility MSD 212/E211 9700 South Cass Avenue Argonne National Laboratory Argonne. IL 60439 USA
Salzburg, 23rd of March 1998, local time: 5.50 p.m.
Dear friends and HistoNetters, is there anyone out there who knows an old ZETOPAN LM (mfctd by = REICHERT, Vienna) equipped with a "double aperture condensor for = bright-field, condensor front lens with an NA 0.95 or NA=3D1.3" , not in = use, stored in a dark cabinet or in the loft ???? (If this question = is/seems silly, please flame privately to my e-mail address) The problem: I use (oh: I "used") a vintage 1964 REICHERT ZETOPAN bright = field LM (YES: unrivalled performance until now) for my routine = semithin section examinations as well as for documenting some of them = since 1981.=20
Last week I (yes, I !) damaged the condensor "deadly" by incident after = having dismounted it for a demonstration of that part in a lecture for = MTA's.=20 LEICA Vienna (formerly REICHERT) told me that (unfortunately) neither a = "new" or a "used" condensor will be available. As I am aware of the = irreparability of that part due to the damaged condensor front lens and = its bent support I only can look forward to the help of the listmembers.
If there is any person knowing of a "hidden", "wrapped and unused" = double aperture condensor for the REICHERT ZETOPAN I should be glad = hearing about "offers" (e.g. to change parts for historical reasons = (e.g. Fluorescence parts), or costs).
Thank you for your help
Wolfgang
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
Dear Hermann, } } Please correct me if I'm wrong, but I seem to remember that, at least some } time ago, all scientific findings and research results, in order to be } considered as valid, should be reproducible by another researcher.
I agree completely. One should be able to reproduce one's own results in any event. That's why it's important to have available the info necessary to go from an initial image to the final one. Another researcher should then be able to replicate the experiment, obtain a similar raw image, process it similarly--not necessarily identically, since the image belonging to the 2nd researcher is not necessarily identical to that of the 1st--and achieve a processed image which demonstrates the 1st researcher's claim. Probably no one will have the time or energy to spend doing this (unless one is studying the same system and wishes to use the 1st researcher's technique), but the scientific community should be convinced that such replication is pos- sible.
} Doesn't the same scheme apply to whatever results that are based on or } supported by image contents? } Yes. I don't see where the fact that the data is in the form of an image invalidates (or changes) the requirement of reproducibility. Yours, Bill Tivol
} ... } } What basis do we use as scientists for deciding what is and is not ethical? } It seems to me that this is not a trivial question, especially when we live } in pluralistic societies and there are a wide range of different approaches } to morality and ethics put forward by different individuals or groups in } society. } ...
I don't think the answer you're looking for can be clearly defined. What might be applied to an image, digital or chemical, runs gamuts between subtle and gross, between merely aethetic to "presentation of an agenda", ... I believe the problem for defining arises out of an infinite number of tools available (especially with regards to digital), most of which allow a significant number of degrees of freedom (especially with regards to chemical).
Whether "we" are laymen or experts, we either have to be aware of the possibilities and ask questions ... as well as not be defensive if questions are asked. Someone else commented on any "manipulation" being repeatable ... I believe this is especially applicable ... it implies the "manipulator" should be able to convey, and that the "questioner" have the tools and knowledge to duplicate and understand. This would also underline how intimate we are with our software ... how many of us really know the algorthym used when their image editting software resamples the image when simply re-sizing for proper presenation?? (... not to imply it is unethical ... but you get the idea ...)
... $0.02 :o)
cheerios, shAf
{\/} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility at the University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
Dear Wolfgang, } } [skip], if the compound is stored appropriate as indicated above and the container is/has been tightened closely every time you used it. And: the amount and how it is stored (and one works with it) makes a difference in the way of acting as a dangerous chemical (with respect to explosions or ignition) } Be careful about the accumulation of dry xtals on the outside of the container. Unscrewing the lid abruptly could cause a big surprise. Yours, Bill Tivol
I like to add the following: One can obtain bacteria by floating a millipore filter on glutaraldehyde solution overnight. Glutaraldehyde sipping through the pores will fix bacteria onto the filter otherwise they may swim/float away.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Agriculture Agri-Food Canada, Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
We find the best way is to use small 0.2um millipore filters. Whatman International do them tel +44 (0)1622 676670 (I have no commercial stake here!)
You can get little ones (13mm) that fit in a special holder that will go on the end of a syringe.
Just squirt your solution of bacteria (probably best washed first in PBS buffer) through the millipore filter (shiny side up) then quickly remove the filter and fix in 3% gluteraldehyde in PBS for an hour or so. wash-pBS osmicate-1% osmium in PBS for 1 hour wash-PBS dehydrate-30%, 70%, 90%, 100%, 100% 20 mins each Then critical point dry your millipore filter (with bugs hopefully still attached) Mount on SEM stub with double sided sticky tape Use conductive paint to make line from surface with bacteria to metal on stub Coat with gold in sputter coater And voila!
Does anyone have any suggestions how to prepare grasshopper antenna and cockroach cerci for fixation, dehyration and embedding. I would like to attempt to thin-section these critters.
Thanks for your suggestions.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
Regarding: } Dear all, } There has been much debate about how to apply ethics to the particular } issue of image processing. I am interested in a slightly more general } question, however. } } What basis do we use as scientists for deciding what is and is not ethical? } It seems to me that this is not a trivial question, especially when we live } in pluralistic societies and there are a wide range of different approaches } to morality and ethics put forward by different individuals or groups in } society. } } } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Ian MacLaren, Tel: (44) (0) 121 414 3447 } IRC in Materials for FAX: (44) (0) 121 414 3441 } High Performance Applications, email: I.MacLaren-at-bham.ac.uk } The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ } Birmingham B15 2TT, } England. } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
There must be some moral and ethical systems which are superior to others, as can be seen from several related lines of evidence: e.g. the existence of the idea of such a superiority of one system over others, practical modern experience that even systems which are supposed to be accepting of all systems of morals and ethics end up being intolerant of some views, etc.
By this line of reasoning, systems which do not accept the possibility of superior systems are self-inconsistent and cannot be such a superior system. We should discard these systems.
A superior system will naturally require that it be imposed on those who do not agree, at least as it relates to their behavior though not necessarily to their belief in the system to which they are subject. This is because of the requirement for self-consistency of such a superior system. Thus, systems which allow a pluralism which accepts "a wide range of different approaches to morality and ethics put forward by different individuals or groups in society" (MacLauren above) cannot be superior systems because they are transparently self-inconsistent. This statement applies very well to the thinking so common in universities today, where many people are terrified of offending anyone by saying that to do some particular thing is immoral or unethical.
What is needed today is a return to Scholasticism, for the kind of foundation for reasoning for which the originator of this question (MacLauren) is looking. Scholastic thinking which has already been set down goes far beyond the kind of argument which I have tried to set up here, and is self-consistent.
And, to provide a concrete link to microscopy, what good is the search for material, scientific truth if we tolerate moral or ethical systems which allow deliberate deception or which see all truth as relative. Scientists just become a group of posturing fools, in search of someting in which they say they don't believe.
} Dear all, } There has been much debate about how to apply ethics to the particular } issue of image processing. I am interested in a slightly more general } question, however. } } What basis do we use as scientists for deciding what is and is not ethical? } It seems to me that this is not a trivial question, especially when we live } in pluralistic societies and there are a wide range of different approaches } to morality and ethics put forward by different individuals or groups in } society. } } } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Ian MacLaren, Tel: (44) (0) 121 414 3447 } IRC in Materials for FAX: (44) (0) 121 414 3441 } High Performance Applications, email: I.MacLaren-at-bham.ac.uk } The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ } Birmingham B15 2TT, } England. } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
****************************************************************************** Douglas Darnowski Department of Crop Sciences 384 ERML 1201 West Gregory Drive University of Illinois Urbana IL 61801 work ph: (217) 244-6150 fx: (217) 333-4777 home ph: (217) 356-6606 fx: (217) 356-4454 email: darnowsk-at-staff.uiuc.edu
We are updating our webpages, including links to Public Domain Software relevant to microscopy, also on FFT's and Image Processing. I would appreciate any suggestions/contributions as to appropriate ones to include. PLEASE respond directly to me, not to the listserver.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Dear all, Not a direct Microscopy question - but it is related in a way. Can anyone help direct me to where I can find the author guidelines for the Journal of Heart Valve Disease? I can't even find out who publishes it - and we do not have it in out Univ. library. I guess you should direct any answers to me at: alexander.black-at-ucg.ie
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Ernest Orlando Berkeley National Laboratory One Cyclotron Road, Berkeley, Calfornia 94720
FOR IMMEDIATE RELEASE
Summer School on Computing in Electron Microscopy slotted for August 17-21, 1998 in Berkeley, California
(Berkeley, CA) The fifth annual Summer School on Computer-Interactive HRTEM Image Acquisition, Processing and Simulation will be held at the National Center for Electron Microscopy (NCEM), Lawrence Berkeley National Laboratory, University of California, Berkeley from August 17 through August 21, 1998.
The curriculum will focus on training participants in techniques of computer-assisted acquisition and interpretation of high-resolution electron microscope images, including remote-control microscopy. Participants will learn general principles and apply them to specific cases. Instruction on use of computer assistance to obtain images on NCEM microscopes will be followed by training in the use of specific application programs for image interpretation by image processing and simulation.
Participants who wish to apply newly acquired techniques to their own projects will be encouraged to extend their visit at NCEM into the next week. Please note: this type of arrangement requires advance submission of a proposal. Projects may involve prepared specimens for microscopy, images and diffraction patterns for processing, or crystal and defect data for simulations. For more information and application materials, contact:
I have a 11 year old Philips CM12. Recently movement of the Objective aperture mechanism either with the apertures in/out or lever right/left causes a pressure rise (to 34) in the column as read by the IGP display. I suspect that movement of the inner most bellows, embedded in the objective lens has developed a micro-leak.
Has anyone else experienced this with their CM12?
Did you try to repair the bellows assembly yourself?
Is their a possibility that the sealing Viton o'ring has gone dry?
What problems can occur with the final alignment of the objective mechanism for the height adjustment?
Did you give up and let the Philips service handle the problem?
**Comments**
If you wish contact can be made privately.
eoptics-at-mcmaster.ca
Thanks in advance
Fred Pearson Electron Optics Coordinator
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
I have a student who is looking for a summer internship in biological TEM and/or SEM in the Milwaukee, Wisconsin area. Does anyone know of any opportunities?
TIA
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
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Brief informational Web focused on Microscopy 10 bit digital / analog CCD cameras http://members.aol.com/dvcco DVC-1300 1300 x 1030 progressive scan pixel array, 12 FPS rate, 8-9 bits out of a 10 bit A/D, RS422 digital out and analog video non-standard for any RS-170 input framegrabber List Price $4995. !!! Other 10 bit 30FPS models available at $2995. Complete frame grabber / software systems available from a one stop US camera manufacturer and systems house. We are an alternative to the other over priced cameras 1K cameras, if the applications fit. This posting is for the benefit of your members.
On Mon, 23 Mar 1998, Fred Pearson wrote: } I have a 11 year old Philips CM12. Recently movement of the Objective } aperture mechanism either with the apertures in/out or lever right/left } causes a pressure rise (to 34) in the column as read by the IGP display. } I suspect that movement of the inner most bellows, embedded in the } objective lens has developed a micro-leak. } } Hi Fred, The commonest cause is the screw at the end of the aperture mechanism being loose. It's worth checking. Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
I will explian you in more detials for your questions in next week when I return home. Due to I have no any document i.e., service manual and circuit diagram in my hand at a moment.
Regards,
Paiboon Nuannin Prince of Songkla University Hatyai 90112 Thailand
On Mon, 23 Mar 1998, Brian McIntyre wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } hi- } } thanks to all who sent info on about testing the vacuum system. } } seems i may have more problems: } } 1) the leak rate from HVac is {3uA/min } atm=250uA } Hvac=25uA on vacuum meter test points (OK) } } 2) V1 doesn't work in auto mode, and many times won't work in manual } } 3) DP heater warms up (light on) in about 20minutes } } 4) R4 vacuum control pot will not make V1 open when vacuum meter {190uA } while in auto mode } } 5) solenoid valve for the closing side of V1 will not release pressure } so that the opening side of the valve (when pressurized) will force the } valve to open.... most of the time (related to #2 above) } } 6) some of the valves (like v4) will actuate manually when the } controls are } in the auto mode (is this normal...or an indication that the logic is } all messed up?) } } 7) V5 (airlock) will not open in auto mode, but does in manual (as } does the } associated leak valve) } } i'm really confused about this since it seems to be a moving target with } many appearent "failures". is there one circumstance or condition that } would explain these observations???? } } thx in advance! } brian } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627 } } 716-275-3058 } 716-244-4936(fax) } } }
====================================================== SECOND INTERNATIONAL SCHOOL AND CONFERENCE ON IMAGE AND FLOW CYTOMETRY =======================================================
Krakow, 07 - 12 June 1998
Jagiellonian University Krakow, Poland
------------------------------------------------------------ Please be so kind as to forward this announcement to your colleagues who might be interested in attending the School
Detailed information: http://helios.mol.uj.edu.pl/ or dobrucki-at-mol.uj.edu.pl USA mirror: http://www.cyto.purdue.edu/mirrors/dobrucki ------------------------------------------------------------
GENERAL INFORMATION ------------------- PLACE: Laboratory of Confocal Microscopy and Image Analysis Department of Biophysics, Institute of Molecular Biology, Jagiellonian University Al. Mickiewicza 3, 31-120 Krakow, Poland
LANGUAGE: English
PARTICIPANTS: The Course is aimed at graduate students, young researchers and clinicians from areas of medicine, biology, biotechnology, chemistry, physics, etc.
LECTURES AND LAB SESSIONS: Participants may attend the lectures only (free of charge) or register for laboratory sessions and tutorials (tuition fee).
GENERAL OUTLINE --------------- The Course will embrace basic and some selected specialized techniques in:
* lectures - open to all students and researchers free of charge, however advance registration required * lababoratory sessions - students will select introductory or advanced level groups. Live cells and fixed specimen will be available for lab sessions; the students are welcome to bring their own samples to be used during lab sessions
INVITED LECTURERS AND TUTORS ---------------------------- E.Bedner, New York Medical College, N.York, USA F.Brakenhoff, University of Amsterdam, Amsterdam, Holland A.Cossarizza, University of Modena, Modena, Italy Z.Darzynkiewicz, New York Medical College, N.York, USA G.Durack, University of Illinois, USA D.P.Evenson, South Dakota State University, USA D.W.Galbraith, University of Arizona, USA W.Goehde, Westfalishe Wilhelms-Universitat, Munster, Germany J.Kawiak, Medical Center for Postgraduate Training, Warsaw N.Opitz, Max-Planck-Institut, Dortmund, Germany A.H.Radbruch, Deutsches Rheuma-Forschungszentrum, Germany J.P.Robinson, Purdue University, Purdue, USA G.Rothe, Klinikum der Universitat Regensburg, Germany F.Sansonetty, IPATIMUP, Porto, Portugal G.Schmitz, Klinikum der Universitat Regensburg, Germany T.V.Shankey, Loyola University Medical Center, Chicago L.Staiano-Coico, Cornell Univ. Medical College, N.York, USA J.Szollosi, Univ. Medical School, Debrecen, Hungary G.Valet, Max-Planck-Inst. fur Biochemie, Martinsried, Germany J.V.Watson, Addenbrooke's Hospital, Cambridge, UK N.White, University of Oxford, Oxford, UK
PRELIMINARY PROGRAMME --------------------- (Lectures, Laboratory Sessions and Tutorials)
* Confocal microscopy - principles and instrumentation * Optimizing data collection in image cytometry * Multiphoton confocal imaging * Principles of image processing and deconvolution of 3D data stacks * Ratio imaging * New fluorescent probes for flow and image cytometry * Green fluorescent protein * Transgene expression * DNA arrays * Accumulation and distribution of antitumor drugs * Chromatin structure in interphase and mitosis * Cytoskeleton and post-translational modifications of tubulin * Plant ploidy and DNA content analysis * Advantages and applications of laser scanning cytometry * Detection of apoptosis and measurement of enzyme kinetic reactions in individual live cells, by laser scanning cytometry
* Flow cytometry - principles and instrumentation * Multidimensional immunophenotyping including analysis of intracellular antigens * Functional T-cell cytometry * Flow Cytometric Analysis of Platelet Activation and Function * Cytokine cytometry * Functional cell studies using fluorogenic probes, fluorescent ligands and FRET * Analysis of activation, proliferation and cell cycle progression * Characterization of apoptosis and necrosis * Apoptosis. Methods of detection and relevance in oncology. * Cytometry detection of cell cycle regulatory proteins * Analysis of rare events including pre-enrichment * Fertility potential of sperm by flow cytometry * Large particle sorting * Magnetic bead cell separation technology
* New frontiers in flow and image cytometry
INSTRUMENTATION --------------- Participants will conduct their experiments using a broad range of instruments used in modern cytometry:
* two confocal microscopes (Bio-Rad MRC1024 and MicroRadiance) * a laser scanning cytometer (CompuCyte) * three Silicon Graphics workstations * three FLOW cytometers, one cell sorter (Becton-Dickinson, Bio-Rad, Partec, Cytomation)
SUGGEST YOUR OWN SUBJECT... --------------------------- The prospective participants are encouraged to suggest subjects of lab sessions that they will find most useful (please contact dr J.Dobrucki).
BRING YOUR OWN SAMPLES... ------------------------- The students will study specimens - live and fixed animal and plant cells prepared especially for the course. All students are also welcome to bring their own samples to be studied. Please contact dr Dobrucki for details.
PRELIMINARY SCHEDULE -------------------- 07 June 19:00 Welcome Reception, registration
08 - 11 June 8:30 - 11:30 Lectures 13:00 - 15:00 Lab sessions and tutorials 16:00 - 18:00 Lab sessions and tutorials
12 June 8:30 - 11:30 Lectures 13:00 - 15:00 Tutorials 16:00 - 17:00 Closing Lecture 19:00 Farewell Party
TUITION FEE ----------- The tuition fee, payable to the Jagiellonian University, is:
* $150,- for graduate students * $200, - for faculty members * $300, - for company representatives
Registration deadline is April 15. After the deadline payment will be $200, $250, $350 respectively.
HOW TO APPLY ------------ Application form is attached at the bottom of this document
STIPENDS -------- A limited number of stipends will be available to participants of the School. Only the students who registered, provided their full cv and a letter of recommendation from their supervisor may apply for financial support or the waver. Please contact dr Dobrucki for details.
COURSE CERTIFICATES -------------------- Those who attended all Course lectures and successfully completed selected lab sessions\tutorials will be entitled to receive a Course Certificate.
SIGHTSEEING AND SOCIAL PROGRAMME -------------------------------- The following trips are planned:
* Museum of the Jagiellonian University (Collegium Maius) featuring a unique collection of scientific equipment * The medieval Salt Mine at Wieliczka. The tourist will visit 30 of 2148 underground chambers * The Tatra Mountains * Krakow Old Town and the Royal Castle of Wawel
We will also organize a trip to Museum of Auschwitz-Birkenau (Oswiecim-Brzezinka) Concentration Camp.
FURTHER INFORMATION ------------------- dr Jerzy Dobrucki dobrucki-at-mol.uj.edu.pl fax. +48 (012) 633 69 07 tel. +48 (012) 634-14-42, ex. 268 tel. +48 (012) 634-16-80, ex. 268
address: Jagiellonian University Institute of Molecular Biology Department of Biophysics Laboratory of Confocal Microscopy and Image Analysis Al. Mickiewicza 3 31-120 Krakow, Poland ============================================================ APPLICATION FORM Second International School of Flow and Image Cytometry Krakow, 07-12 June 1998 -------------------------------------------------------------- Send to: DR J.DOBRUCKI JAGIELLONIAN UNIVERSITY, INSTITUTE OF MOLECULAR BIOLOGY AL. MICKIEWICZA 3, 31-120 KRAKOW, POLAND fax: +48 (012) 633 69 07 e-mail: dobrucki-at-mol.uj.edu.pl -------------------------------------------------------------- Please type in using CAPITAL LETTERS
First Name:................................. Family Name:................................. Occupation (undergraduate, graduate, post-doc, faculty, other):.................................. Institution: full name............................ postal address:........................ telephone............................... fax..................................... electronic mail......................... Name and address of your supervisor, group leader or laboratory director:..................................... ........................................... ........................................... I want to attend laboratory sessions on (please mark with X): flow cytometry - introductory...... flow cytometry - advanced...... image cytometry - introductory...... image cytometry - advanced...... laser scanning cytometry - ........
I suggest the following additional topics for tutorials/practicals: 1............................................... 2............................................... 3............................................... 4............................................... ...............
Please enclose your curriculum vitae, including a list of publications. The tuition payment should be sent by bank transfer, and a copy of the bank document must be sent by fax to Dr. J.Dobrucki, fax +48 (012) 633 69 07.7.
I have been asked by Dr. John E. Johnson, Jr., Editor-in-Chief of Microscopy Research and Technique (MRT), to solicit selected articles addressing recent advances in our understanding of DIGITAL IMAGING. The invited articles would be the basis for a topical issue of MRT entitled DIGITAL IMAGING IN MICROSCOPY, on which I would serve as Guest Editor. Possible topics could be: Theory of digitization of micrographs and digital imaging. Evaluation and comparison of scanners, CCD array detectors, imaging plates etc. Sampling and undersampling phenomena (and their avoidance or application). Application of CCD arrays in imaging, diffraction, automated microscopy etc. Measurement and correction of transfer functions. Digital image processing of micrographs. Data bases and image retrieval. etc. In this regard, I am writing to invite you, either alone or in conjunction with a collaborator(s) of your choice, to submit a suitable article. A manuscript length limitation is set at about 30 double-spaced typewritten pages, plus micrographs. An Abstract should be included. Each manuscript will be refereed, and therefore will be suitable for inclusion in Grant Proposals or Renewals. The MRT Instructions to Contributors at http://www.interscience.wiley.com/jpages/1059-910X/authors.html should be used as a guide when preparing the manuscript. Note that the journal has a format of 8 1/4" by 11", and the figure size is 6 3/4" wide by a maximum of 9" high for a full page width figure, or 3 5/16" wide by a maximum of 9" high for a single column width figure. Therefore, your figures should be trimmed to these dimensions if you wish them to be reproduced at the same size as the originals. There is a charge to authors for color figures, but no charge for black and white figures, regardless of the number. You would need to include copyright release forms signed by the publisher for any previously published figures. Please examine MRT for examples of what we are looking for in topical papers. If you would be willing to participate in this venture, I would appreciate knowing your intentions by 30.4.1998. I would need your Article Outline (with approximate number of pages and figures) by 30.6.1998, and I would, at that time, make available to you a list of other contributors and article titles. The completed manuscript would be due in my office by 30.8.1998. All manuscripts are peer reviewed and may require revision. The topical issue will be published within 4 - 6 months (this is part of our agreement) after the manuscripts are received at the publisher's office in New York (John Wiley). I hope you will agree to contribute to what we believe will be a fascinating and beneficial update of current knowledge in DIGITAL MICROSCOPY.
Sincerely,
Philip Koeck
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
One of my big concerns about digitally enhancing images is the tendency for enhancing an image so much to support one's point, that the image is no longer photo-quality, but merely a cartoon. I am aware that when people selectively crop their photos, and zoom in on only areas of interest, a lot of information is slanted to support a point, but I don't want these things taken to extreme. If one starts cloning away background label that they don't think belongs, or adding color to images to highlight the cells or structures of interest, then I seriously begin to question the objective value of this information. (Reproducible or not!)
I guess this is my attempt to 'get on my soapbox' and implore you all to use digital enhancement with temperance!
Thanks for listening.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan, USA
---------- } Van: yvan lindekens {yvan.lindekens-at-skynet.be} } Aan: darnowsk-at-staff.uiuc.edu } Onderwerp: Information about Lensman } Datum: dinsdag 17 maart 1998 18:27 } Mr. Darnowski,
The Lensman Microscope is made by Science of Cambridge ltd. (U.K.).
* Lensman1,2,3.jpg: article about the Lensman microscope in the German magazine "Mikrokosmos", mai 1992.
You can also find some articles about the Lensman and other portable microscopes, including the "McArthur microscope" in:
http://www.microscopy-uk/mag/libindex.html
I have used the Lensman for a few days, and I find the results rather dissapointing: image quality is poor, only slightly better than the quality of the image produced by a toy microscope!
I have had better results with a MBU-4S microscope (ZENIT/LOMO, Russia) (see Spiro.jpg).
This microscope costs in my country about 8 000 Belgian Francs (that's about 213 US$).
Zenit/LOMO is availlable in the USA, but don't ask me who the dealer is: I don't know...
Of course, this is a "real microscope": it's not intended to be a "portable", but in my experience it can very well be used as such. It's weight is about 3 kg. It's equipped with a good abb=E9 condenser, and objectives 8x, N.A. 0,20 and 20x, N.A. 0,40; Oculars 7x H, 10x K and 15xK. When I use it, I put it, in a large towel, in my bag...
* spiro.jpg: conjugation of Spirogyra sp. photographed "in the field'" with natural light; obj. 20x, oc.: 7xH. With camera Olympus OM1 and film Ilford Pan F.
Photographic print Scanned with 200x200dpi.
I have had also good results with a portable microscope "AZ 2". This is also a Eastern-European microscope, made by MEOPTA. It was, a few years ago, availlable trough "KOSMOS SERVICE", Pfizerstrasse 5-7, 7000 Stuttgart1, Germany. Perhaps it is still availlable...
I have not forwarded the attachments. Anyone wanting them should email me personally--Doug Darnowski
****************************************************************************= ** Douglas Darnowski Department of Crop Sciences 384 ERML 1201 West Gregory Drive University of Illinois Urbana IL 61801 work ph: (217) 244-6150 fx: (217) 333-4777 home ph: (217) 356-6606 fx: (217) 356-4454 email: darnowsk-at-staff.uiuc.edu
First, thank you to everyone who sent EDS upgrade suggestions, it has been enormously helpful.
Please remember that the following is only a summary of the responses that I received to my inquiry about an inexpensive upgrade for my old Kevex 7000, it is not a comprehensive survey.
In my original message I mentioned that I was considering the 4pi Spectral Engine II package. Of the 8 users who responded, 5 wrote to say they liked the 4pi system a lot. They said that they thought it represented the best value for the money, the tech support was good and they liked the DTSA and Flame software. Other users recommended the IXRF, Rontec, and EMispec systems. The IXRF is a more comprehensive and expensive system than I'm looking for as is the Rontec system that is available in the US. I was unable to find additional information about the EMispec.
I also heard from a number of vendors. For the most part the upgrade packages they offer include electronics such as the pulse processor (which for reasons of cost I prefer not to replace) and proprietary software. In general the prices were in the neighborhood of $30K. The exceptions are Scanners Corp., Mektech Inc., E.Fjeld Co., and American Nuclear Systems Inc. which all offer relatively inexpensive keep-your-own-electronics upgrades. I have an assortment of addresses and quotes I'd be happy to share with other interested users.
Our materials science lab has recently inherited a Nikon Fluophot transmitted fluorescence microscope. HELP!! Having no experience with fluorescence, we would appreciate any hints specific to this "dinosaur". Thank you. Tina McNickle University of Connecticut IMS Microscopy Laboratory 97 North Eagleville Road Storrs, CT 06269-3136 Phone (860)486-5453
The Department of Anatomy & Cell Biology at the University of Michigan has a Meridian Ultima 312 Premium Laser Confocal Microscope System for sale. This instrument was purchased with private funds in 1996 by a faculty member and was used for six months to meet the requirements of the research contract. After that, it was placed in a multi-user facility where it saw very little use. It is in perfect condition and was recently inspected by the parent service company. A field service report of this inspection is available. Space is ultimately the issue since the multi-user facility (CBL) is slated to be renovated and ultimately downsized. The original price of this instrument was $382,110.
This system has a host of desirable features including an Innova Enterprise UV-Visible laser (351-364 nm, 488nm, & 514 nm) and a Krypton laser (568nm & 647nm) with filters for Indo, FITC, Rhodamine, PI, BCECF, etc. The microscope body is an Olympus IMT-2 with 4X, 10X, 20X, 40X, 60X, and 100X objectives. It features mirror and stage scanning operations, nine different aperture/pinhole sizes, motorized stage, 3 PMT's, and "Z" drive. An optional software package is included which contains functions such as: Image Analysis, Cell-Cell Communications, FRAP, List, Sort, Auto Image, Ratio Analysis (calcium & pH), and Kinetics Analysis. The computer configuration is a 90 MHz EISA/PCI Pentium with 80 MB RAM, VGA Graphics, 2.1 GB SCSI hard drive, two monitors, and 1.2 GB rewritable optical drive.
Interested investigators are invited to contact us directly at 734-763-1170 during normal business hours or through a personal e-mail response. Thank you.
} Our materials science lab has recently inherited a Nikon Fluophot } transmitted fluorescence microscope. HELP!! Having no experience } with fluorescence, we would appreciate any hints specific to this } "dinosaur". Thank you.
It dates from the late '70s and is adaptable to reflected as well as transmission fluorescence. Uses standard Nikon optics.
Who out there are the people in the know regarding remote imaging. Can I send live / semi live images to a person while we are both on the phone and discuss what I am doing on the TEM? We are currently using a Kodak Megaplus, into a computer with a VGA video feed out to the monitor. I have already played with converting that feed to TV video, and recording a session on a VCR (with surprisingly good image quality). I would like to know how to make the next jump.
I am planning on viewing some thin sections of cattle grub spermathecae and cattle grub testis and I am unable to find previous research done on these specimens with regards to fixation, dehydration, and embedding. Does anyone out there know of any method which might work best? Suggestions would be very appreciated.
Thanks very much for the suggestions.
Paolo Santangelo University of Lethbridge Lerthbridge, Alberta Canada E-mail - santpe-at-uleth.ca
} } Who out there are the people in the know regarding remote imaging. ------cut------ } I } would like to know how to make the next jump. } } Rick Vaughn
There is a major state-of-the-art program in TelePresence Microscopy which is funded under the DoE 2000 Program, called the Materials MicroCharacterization Collaboratory. That project connects 5 national electron-optical beam characterization centers (ANL, LBNL, NIST, ORNL, and Univ. of Illinois) into an on-line virtual electronic collaboratory.
The MMC deals with far more than just simple imaging over the Net, it's objective is to integrate (ultimately) all the methodologiew which defines scientific collaboration (remote control, data sharing, electronic notebooks, video conferencing, etc....). You can find out more about this project at the URL
http://tpm.amc.anl.gov/mmc
You can also login to the "live" multi-site collaboratory (using NetScape not MS I.E.) at the following:
http://tpm.amc.anl.gov/mmc/TPMLVideoCollab.html
} From this site you can connect also directly to individual facilities within the Collaboratory.
There is also a Major Symposium on the same topic at the upcoming Microscopy & Microanalysis '98 Meeting in Atlanta this July
the Symposium is entitled: Advances in Remote Microscopy, Instrument Automation and Data Storage and is Organized by: Edgar Voelkl, Mike O'Keefe and Nestor Zaluzec . At MM'98 you will have the opportunity to hear the latest advances in TelePresence Microscopy & Microanalysis from a wide range of perspectives covering both the Physical and Life Science arena's. There is a full day of platform presentations as well as an extensive poster session, if all goes well there should also be the opportunity for on-line demostrations at the annual computer workshop.
Cheers...
Nestor Your Friendly Neighborhood SysOp
************************************************************** Disclaimer: I have an modicum of vested interest in both the MMC and the MM'98 Program **************************************************************
Molecular Imaging Spring Workshop on Biological High Resolution In Vitro Atomic Force Microscopy (nanometer resolution of soft samples in biological buffers at 37C) Hands on training. Bring your own samples (NO BIO HAZARD MATERIALS)
I am the senior anatomopathologist of thePaediatric Institute of Research of the University of Trieste(Italy).Due to two international collaborative research projects we ought to send to UK and USA our frozen section of intestinal biopsies. These sections will be analyzed for EmA(antibodies anti-endomisium) in Celiac Disease with immunofluorescenze indirect (USA - Baltimora) and for cytokines TNF-alfa in IBD with ibridation in situ (UK). I fix the frozen sections with acetone-clhorophormio and store them at -20=B0C (they last 2-3 months). I would like to know if ther is any other tecnique to fix sections in way I could store at 2-4=B0C and preserve them before searching for antigen for a longer period (such as 5-6 months).Thank you Dr. Angelo Citt=E0 e-mail: acitt-at-tin.it
I mistiped a number and now I am lucky owner of 8000 Gilder GS2x1 slot grids, made of Cu. Are there out some other TEM users haveing interest in purchasing such things?
(I have strong commercial interest in getting away these grids!)
Best regards Peter
dr Peter Nagy MTA MFA 1525 Budapest POBox 49 T: ++36-1-3959220/1968 e-mail: p.nagy-at-r1.atki.kfki.hu
} I am planning on viewing some thin sections of cattle grub spermathecae } and cattle grub testis and I am unable to find previous research done on } these specimens with regards to fixation, dehydration, and embedding. } Does anyone out there know of any method which might work best? } Suggestions would be very appreciated.
This tissue is not difficult to prepare, particularly if there is no chitin around. I suggest that you use any reliable protocol for preparation of animal tissues for TEM. If you like you can refer to a protocol that I published a few years ago. I know that it works very well for insect material. The reference is:
Cross, R.H.M. (1989) A reliable epoxy resin mixture and its application in routine biological transmission electron microscopy. Micron & Microscopica Acta, 20(1), 1.
If you do not have this journal available I could mail or fax the reprint to you.
Good luck!
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377
To: microscopy-at-sparc5.microscopy.com -at- Internet-Mail cc:
I would like to remind anyone who may be interested in the following= position that the application deadline is April 10, 1998. If there= are any questions, please contact me at laura.rhoads-at-wku.edu. Thank= you.
EM TECHNICIAN- The Department of Biology at Western Kentucky University= is accepting applications for a full-time, permanent Electron Microscopy= Technician position available July 1, 1998. The major responsibility= of this position is to oversee activities associated with the= multi-disciplinary user EM Facility. The Facility houses two JEOL= 100B TEMs, one JEOL 5400LV SEM with EDS, a darkroom, and biological= sample preparation equipment. The technician will be responsible= for the day-to-day operations of the Facility including maintenance= of the instruments and trouble-shooting, sample preparations, inventory= of equipment and supplies, training of users, and providing research= support for faculty, students, and outside users of the Facility.= The person hired will be encouraged to assist in obtaining outside= support for the Facility through contracts. The technician will= report directly to the Facility Director. The qualified candidate= will have a B.S. degree (M. S. preferred) and at least two years= experience in aspects of electron microscopy. Additional duties= will be assigned and may include supervising laboratory preparations= for cell and molecular biology courses and full support for the= darkroom. Send letter of application, curriculum vitae and three= letters of reference to: Dr. Laura S. Rhoads, Department of Biology,= Western Kentucky University, 1 Big Red Way, Bowling Green, KY 42101-3576.= Screening of applications will begin April 10, 1998. Questions may= be addressed to laura.rhoads-at-wku.edu. Women and minorities are especially= encouraged to apply. WKU is an affirmative action/equal opportunity= employer.
************************************************************ It's true- the inmates ARE running the asylum... ************************************************************ Laura Rhoads Electron Microscopy Facility Director Department of Biology Western Kentucky University 1 Big Red Way Bowling Green, KY 42101-3576
My name is Andrew Aurbach. I am currently developing a video project with National Geographic Television. I am seeking moving microscopy images at the cellular and atomic levels. I am curiuous at to some of the materials which may be available at the Association level. I checked some of the videotape resources, and some may be appropriate. Who can I contact on this front?
When your message arrived, I was reading and article titled "Desktop Microscopy Apps Get Their Video-Rate Imaging" in the March 1998 issue of Advanced Imaging magazine (p. 18). The article/advertisement highlighted particle tracking systems for biological applications.
The author can be reached at: Dr. Douglas M. Benson Inovision Corp. Durham, NC (919) 361-4609 http://www.inovis.com
The usual disclaimers apply. I have no financial interest in products or companies mentioned in this message.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
I am looking for advice on doing TEM on a 20 micron thick sheet of polypropylene film. There is a small dark threadlike inclusion that is buried in the matrix that I would like to identify using EDS. Does anyone have experience with this type of analysis? Thank you in advance.
Regards,
Michael Coviello EM Lab Manager Materials Science Dept. The University of Texas -at- Arlington Arlington, TX E-mail coviello-at-mae.uta.edu 817-272-5496
One of my colleagues asked me to do some TEM on a "nuclear pellet", cells from adult rat cerebral cortex that had been fractionated and the nulcei seperated out. The material looks relatively 'clean' but it would be nice if I had a reference to cite. Can anyone help me out? Thanks in advance.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Meeting of the Metropolitan Microscopy Society ==================================================
Date: Wed., April 1, 1998 Time: 10:00 AM Place: Raddison Inn, 601 From Road, Paramus, NJ Directions: Garden State Parkway to Exit 165 (E. Ridgewood Ave./Oradell Ave.) From Road is on the west side of the Garden State Parkway, and adjacent to it.
10:00-10:30 am Registration: ($5.00) Coffee and Danish sponsored by: IBM Analytical Services Hopewell Junction, NY 12533.
10:30-10:50 am Introductory remarks and society business -- Phil Flaitz.
10:50-11:40 am PRINCIPLE AND APPLICATIONS OF ELECTRON BACKSCATTER DIFFRACTION (EBSD) IN THE SEM, Prof. Frederic Cosandey, Department of Ceramic and Materials Engineering, Rutgers University, Piscataway, NY
The diffraction phenomena known as Electron Backscatter Diffraction (EBSD) is known since the 1930's but is only recently that EBSD has become a practical technique for materials characterization in a Scanning Electron Microscope (SEM). These recent developments of EBSD have been made possible by the availability of fast PC computers for automated diffraction pattern recognition as well as for automated beam control and automated analysis of over thousand of patterns in a reasonable amount of time. There are now numerous application of EBSD ranging from phase identification to texture analysis, orientation imaging microscopy and strain analysis in metal, oxides, semiconductor and composite materials. In this talk, the history and basic principle of EBSD will be reviewed with a discussion on spatial resolution limit and range of applications. Specific examples will be presented which include epitaxial orientation determination of Au particles on TiO2, orientation and perfection determination of oxide crystals and texture analysis in Al2O3 polycrystal.
11:45-12:45 pm STANDARDS FOR X-RAY MICROANALYSIS: HOW WE GET THEM, HOW WE USE THEM AND WHY WE WILL ALWAYS NEED THEM, Greg Meeker, United States Geological Society, Denver, CO.
X-ray microanalysis, whether by energy or wavelangth dispersive meth- ods, must rely on standards for accuracy and precision. Standards provide the means of calibrating our instruments and verifing that our results are acceptable. Even standardless methods require the use of well characterized calibration materials.
With the large number of EDS and WDS systems in use today it is surprising that very few certified microanalytical reference materials are available. This is particularly true of non-metallic standards such as oxides, sulfides and semi-conductors. There are many natural and synthetic materials that are used as standards throughout the world for x-ray microanalysis, but it is not always clear how to obtain or prepare these materials. In acquiring or preparing standards several factors must be considered including homogeneity at the micrometer scale, stability of the material under analytical conditions, accuracy of reported compositions, and uniformity of x-ray emission with respect to orientation of the material. It is also not always clear how to choose the appropriate standard for a particular element since the matrices of both the standard and the unknown can significantly effect the measured intensities. This is particularly true with the ultra-light elements where variation of peak shape due to chemical bonding, and the uncertainty of published mass absorption coefficients become critical.
Another factor effecting how we use and prepare standards is the recent move toward stricter quality assurance practices and requirements, such as ISO 9000, and the actions of international standards committee. These changes present new and consequential challenges and concerns for the analyst.
These subjects will be addressed, and suggestions will be presented for obtaining and using reference materials that are presently available. In addition, a brief summary of recent efforts at the U.S. Geological Survey to produce a relatively large quantity of a basalt glass standard suitable for microanalysis will be presented. Finally, the present situation with respect to ISO and the International Standards Committee, and the effect these groups could have on our procedures and practices as analysts will be discussed.
12:45 pm Lunch. (Available at Max's Cafe in the Radisson Inn)
For additional information, please contact: Phil Flaitz ..... 914-892-3094, flaitz-at-us.ibm.com Jeff Hurd ....... 914-894-4250, HURDJ-at-US.IBM.COM
Philip L. Flaitz IBM Analytical Services Ph.......(914) 892-3094, FAX -2003 flaitz-at-us.ibm.com
As someone said, it was serendipitous. I was processing a bunch of images through Visilog 4.x running on a Windows box as I read your letter.
There is a SAVE option which allows several different formats. I see TIFF and TGA (Targa?) as options in addition to a couple of proprietary Visilog formats. I don't know if they do 16-bit TIFF, but they certainly should do 8-bit unless you have a really old version. I know Visilog is now out in version 5.x. But we are sonstrained to version 4 for a while longer as it is bundled into our EDS package.
At 09:29 AM 3/25/98 -0500, you wrote: } Hello All, } We are running Cameca's Visiwiew (Noesis' Visilog) to display and process } images and x-ray maps acquired on a Cameca SX-50. The program is running on } a Sun Unix platform under Sunview v3.0 environment. Up to now, we } converted images to other formats by using a unix screendump utility to } convert the screen displayed images to sun rasterfile images which are } readable by many second party image processors. The conversions, however, } were limited to display resolutions only. I would like to convert 1024 X } 1024(16 bit deep) visiview images to other formats(TIFF, PICT, etc.) and } preserve the original resolution. Any advice will be greatly appreciated. } } Regards, } Mark S. Angelone
To keep millipore filters flat during tissue processing and CPD, I have sandwiched the filters between doughnut-shaped magnets, which are approximately the same size as the filters. It works great. Good luck.
Dear Mike, I've done something similar with a polymer. I spoke nicely to a zoologist who cut thin sections with a diamond knife and picked them up on copper grids. Then I coated the sections with carbon - if you don't do this the charging will tear the polymer apart. The rest is easy; just spend hours looking for the inclusion and try not to search in the resin the zoologist used to make the block for sectioning.
Regards, Eric On Wed, 25 Mar 1998 09:20:06 -0600 Mike Coviello {Coviello-at-mae.uta.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Microscopists: } } I am looking for advice on doing TEM on a 20 micron thick sheet of } polypropylene film. There is a small dark threadlike inclusion that is buried } in the matrix that I would like to identify using EDS. Does anyone have } experience with this type of analysis? Thank you in advance. } } Regards, } } } Michael Coviello } EM Lab Manager } Materials Science Dept. } The University of Texas -at- Arlington } Arlington, TX } E-mail coviello-at-mae.uta.edu } 817-272-5496 } }
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
The Mountain States Society of Electron Microscopists and the Colorado Microbeam Analysis Society, a local affiliate society of the Microscopy Society of America and the Microbeam Analysis Society, will hold it's annual Spring Symposium in Boulder, CO, on Friday, April 17. Details about the program and location are available at their new website:
Michael Coviello wrote: =================================================== I am looking for advice on doing TEM on a 20 micron thick sheet of polypropylene film. There is a small dark threadlike inclusion that is buried in the matrix that I would like to identify using EDS. Does anyone have experience with this type of analysis? Thank you in advance. =================================================== This is a fairly common kind of industrial problem of the type that arises in a polypropylene film extrusion line and the analysis if pretty much routine. At 20 um film thickness, this is really very thin, so it clearly has to be embedded. The sectioning must be done cryo, and we would also recommend gold sputter coating one side and aluminun evaporation on the other, if the film is assymetric so that you can always readily keep track of the two surfaces. Knowing which side is which is very often an important bit of information to have. We have had excellent results using our own SPI -Pon(TM) 812 embedding resin kit but some of the other "Epon(R) replacements" would (probably) work just as well. Diamond knife sectioning is preferred over glass knife sectioning. If the "thread" is a "line up" of catalyst reside particles (quite possible in the case of polypropylene if the screen pack filters in the extrusion lines are failing), then you might just as well use a materials science diamond knife and not risk the destruction of a perfectly fine (and extraordinarily expensive in comparision) life science diamond knife. Sometimes it is difficult to get SAED from the catalyst reside particles but getting EDS data is no problem, so long as you have made stable enough sections(e.g. they are thin enough).
Note: There is another approach that in some instances is easier and faster for this kind of very thin sample: Use a plasma etcher to etch away the polypropylene down to the point you hit something that won't etch. Or else, etch to completion, until there is just a residue that is left on the substrate, say a beryllium planchett. That could be your "thread" of interest. Now this is the ideal kind of SEM/EDS sample and you can spare yourself the grief and aggrivation of trying to thinsection this kind of sample altogether. Of course if the "thread" itself etches away, that tells you something as well, namely it is carbonaceous, maybe a carbon black contamination problem but certainly not a catalyst residue kind of problem. Otherwise something would be left after the etching. Using a Be planchett really is the ideal kind fo substrate because of its very low Bremmstrahlung background radiation levels.
Hope this helps.
Disclaimer: Our firm performs these kinds of analyses for clients as an independent laboratory service. We are also suppliers of diamond knives and embedding resins for this kind of work. And if the SEM approach is used, we also manufacture the plasma etcher and supply the Be planchetts.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I'm looking for a copy of the February 23, 1998 TIME magazine (vol.151, no.7). It's the one with the cover of a negative stain of the Hong Kong chicken/human flu virus & a scientist in a "bunny suit". We're going to have our lab available during campus open house & thought that the article, picture, etc. was a good way to show the general population how TEM's help us in the real world. If anyone knows of a source or has one hanging around that they no longer want, please let me know. Also, if there is a location where we can find pictures of Ebola or any other viruses please let me know. We think that most people know what viruses are so we'd show them instead of trying to explain what the inside parts of a cell are.
We've never had open house before, so if y'all have any cool suggestions for displays send 'em my way.
Thanks Kiddos!
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
A postdoctoral position is open at Northwestern University to work using Ultra-High Vacuum Electron Microscopy of Surfaces, and Direct Methods for surface structure determination using electron and x-ray diffraction data. Some details about the available hardware can be found in http://www.numis.nwu.edu, and see also Collazo et al, Phys Rev Letts 80, 1678 (1998). A strong background in electron microscopy is required, and additional background in UHV will be highly relevant. Interested applicants should send a short CV via email to ldm3-at-apollo.numis.nwu.edu including email addresses for references.
++++++++++++++++++++++++++++++++++++++++++++++++ Prof. Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 email:ldm3-at-apollo.numis.nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Hi Todd: I worked on PLANT sperm cells using SEM and immunogold labeling of myosin on sperm surface . I used formvar coated nickel grid (pre-coated with carbon) to attach the sperm cells (need to centrifuge for a while). The sample was fixed with 1% glutaraldehye and then dehydrated with ethanol, CPD and coated with carbon. It turned out pretty good. Hope it'll help.
Zhaojie Zhang Department of Botany and Microbiology University of Oklahoma Norman, OK 73019 405-325-6234
Todd Rippy wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am looking for a Fixation protocol for a mouse fertilization study. } Any leads on similar work would be appreciated. } } There is an SEM of rat fertilization in a text credited to D. M. } Phillips of Visuals Unlimited. I have searched with no success for } this company. } } Thanks, } Todd Rippy
Geoff, be happy and congratulate your collegue for the purity of his fraction. Many a sample of organelles biochemists deliver for TEM-screening are highly contaminated with other particles (if they contain the desired fraction at all!). Thus, I am afraid there is hardly any "standard" published which you could cite as reference. It depends on the biochemists skill.
Greetings, Norbert; Frankfurt, Germany
} ---------- } Von: Geoff McAuliffe[SMTP:mcauliff-at-UMDNJ.EDU] } Antwort an: mcauliff-at-UMDNJ.EDU } Gesendet: Mittwoch, 25. M=E4rz 1998 20:13 } An: Microscopy-at-Sparc5.Microscopy.Com } Betreff: nuclear pellet } =20 } = ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America=20 } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } = ---------------------------------------------------------------------- } -. } =20 } Dear List: } =20 } One of my colleagues asked me to do some TEM on a "nuclear } pellet", } cells from adult rat cerebral cortex that had been fractionated and } the } nulcei seperated out. The material looks relatively 'clean' but it } would } be nice if I had a reference to cite. Can anyone help me out? Thanks } in } advance.=20 } =20 } Geoff } --=20 } *************************************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane Piscataway, NJ 08854 } voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu } *************************************************************** } =20
THE GOLGI COMPLEX. State of the art 100 years after Camillo Golgi's discovery
Pavia, Italy . September 19-23, 1998
Organized by
San Matteo Hospital, University of Pavia, National Research Council, Mario Negri Sud Institute
Organizing Committee: De Matteis (Italy); K.E. Howell (USA); A. Luini (Italy) and A.A. Mironov (Italy)
Scientific Adversary Committee: F. Clementi (Italy); M.G. Farquhar (USA); G.E. Palade (USA), K. Simons (Germany) and G. Warren (UK).
Local Committee E. Solcia (San Matteo H., Pavia), A. Calligaro (Univ. Pavia), S. Riva (CNR, Pavia)
Topics - Architecture of the Golgi complex and composition of the Golgi compartments. - Transport from the endoplasmic reticulum to the Golgi; - Intra-Golgi transport; - Transport from the Golgi to the plasma membrane. - Plasticity and inheritance of the Golgi complex. - The cytoskeleton and the Golgi complex. - Classical and alternative intracellular traffic models. - The role of signaling in Golgi dynamics. - Primary events in Golgi dynamics: vesicle budding and fission, membrane fusion, membrane tubulation.
Speakers include: W. E. Balch, USA; V. A. Bankaitis , USA; J. J. M. Bergeron, Canada; J. S. Bonifacino, USA; N. Borghese, Italy; P. De Camilli, USA; S. Emr, USA; M.G. Farquhar, USA; B. Goud, France; H.-P. Hauri, Switzerland; J. Heuser, USA; W. Hong, Singapore; K. E. Howell, USA; T. E. Kreis, Switzerland; J. Lippincott-Schwartz, USA; V. Malhotra, USA; P. Melancon, USA; J. Meldolesi, Italy; D. J. Morre, USA; J. Morrow, USA; S. Munro, UK; P. J. Novick, USA; G. E. Palade, USA; H. Pelham, UK; A. Rambourg, France; J. Rothman, USA; R. Schekman, USA; K. Simons, Germany; L. A. Staehelin, USA; G. van Meer, The Netherlands; M. Vaughan, USA; G. Warren, UK; M. G. Waters, USA; F. Wieland, Germany; P. Weidman, USA; M. Zerial, Germany
Registration Deadline: May, 15th, 1998
The total ,umber of participants will be limited to 200. There will be poster session. The Conference fee of 600 dollars covers accomodation in student colleges (Residenza Golgi and Collegio Castiglioni) in single rooms (with bathroom) lunch, registration fee and group transportation.
Application and abstract forms from Anna Cavallo: e-mail: cavallo-at-cmns.mnegri.it
For further information contact:
Dr. A. Mironov, Consorzio Mario Negri Sud, Via Nazionale, S. Maria Imbaro (Chieti), 66030, Italy, Tel. +39 872 570 323, Fax. +39 578 240, E-mail: mironov-at-cmns.mnegri.it Internet: www.cmns.mnegri.it/golgi/
} I am looking for advice on doing TEM on a 20 micron thick sheet of } polypropylene film. There is a small dark threadlike inclusion that is buried } in the matrix that I would like to identify using EDS. Does anyone have } experience with this type of analysis? Thank you in advance.
Most of the ideas already received are very good. But you may not have certain bits of apparatus available, so here is our history of a similar problem:
We have also looked at "things" inside polypropylene films. The first thing is, do you have any idea whether this object is organic or inorganic?
If you don't know which, then it would seem that methods based on cutting, which will not chemically attack the "inclusion", are best.
If it's inorganic, then things are very much easier. Charles Garber's plasma etching idea sounds just the ticket, but where you don't have such apparatus available, you could try dissolving away the PP. We have done something like this:
Stacked inside a covered dish in an oven at 120^C:
- Specimen film - Regenerated Cellulose membrane filter - Glass fibre pad (Whatman GF series)
standing in a layer of decalin, which will slowly dissolve away the polypropylene, leaving the inclusions on the membrane filter.
We have also used permanganic etching to remove so many microns of the film. The film could be stuck down to a polystyrene or similar sheet, to keep it flat. After etching you can look under a reflection optical microscope to see if the inclusions have come to the surface. You can find our publications by going to my home page (URL below) and clicking on "Publications" in the menu bar at the top - the ones relevant to etching are in the first section of that page.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
you asked for suggestions for displays. A lot depends on the age group that you expect to get but I have found that from ages of 11 upwards it is nice to show something on a light microscope and also on the electron microscope (you haven't said whether your visitors will get to drive anything). A nice motile bug such as Rhodospirillum works well because your visitors can see it small and moving on the light microscope (we displayed it on a TV monitor) and big with flagella on the TEM.
We also gave out little stickers (badges would have been better) saying "I drove the ****** electron microscope at ******* and the kids loved it - they were nearly as popular as balloons. The most popular pictures that we gave away were of disease causing organisms such as influenza. I suspect that a couple of short videos/slides/multimedia displays running on monitors would grab attention more than just static photos.
The last time we did this we were busy all day so make sure you have enough people to help out and take breaks and good luck.
Malcolm Haswell Electron Microscope Unit University of Sunderland UK ----------
Websters,
I'm looking for a copy of the February 23, 1998 TIME magazine (vol.151, no.7). It's the one with the cover of a negative stain of the Hong Kong chicken/human flu virus & a scientist in a "bunny suit". We're going to have our lab available during campus open house & thought that the article, picture, etc. was a good way to show the general population how TEM's help us in the real world. If anyone knows of a source or has one hanging around that they no longer want, please let me know. Also, if there is a location where we can find pictures of Ebola or any other viruses please let me know. We think that most people know what viruses are so we'd show them instead of trying to explain what the inside parts of a cell are.
We've never had open house before, so if y'all have any cool suggestions for displays send 'em my way.
Thanks Kiddos!
Paula :-) Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Help! I'll be receiving some resin beads with cell organelles (presumably) attached soon and I've never dealt with this sort of prep before. I would appreciate hearing from anyone out there with helpful information about the processing, handling etc. of these things. I've been off the listserver for a while so if it's already been discussed, please just answer privately. Many thanks. Grace
Thank you for quick responses (MFM and Z.Z). The question that I posted was rather vague, but your response has been helpful. We are working on the
Do you know of a published version of this protocol?
I am concerned with issues of the isotonicity of the buffer/glutaraldehyde; timing of fixation after the fertilization; effects/artifacts of removing the zona pellucida.
I am trying to find information on the number of optical microscopes currently in use, preferably by category such as hospital, research, university, U.S., world, perhaps binocular/monocular, etc.
Anybody have a suggestion where I might go search?
People always like insects, spiders, etc. in the SEM. Especially when they're put in live, imaged, then release live.
If you really want to have fun, put in house dust mites, or eyebrow mites (Demodex folliculorum, if I remember right). The latter are soft-bodied beasties and require carefull drying so the opisthosoma doesn't flatten and wrinkle to oblivion, but the reactions are worth the trouble. 8-)
Phil
} We've never had open house before, so if y'all have any cool } suggestions for displays send 'em my way. } } Thanks Kiddos! } } } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 5037 Station A Champaign, IL 61825-5037 (217) 355-1143 oshel-at-shout.net or poshel-at-hotmail.com ***** looking for a job *****
For anyone doing negative stains....where do you get viruses to use for size standards? There is a protocol in one of the Hayat books for getting TMV from cigarette tobacco - has anyone tried it? Do you think all cigs are contaminated with TMV, or just the cheap-o brands?
Sorry if this question seems frivolous, but I DO need a size standard, and I'd prefer a "biological" one to beads or crystals. Virus seemed most logical :)
Is anyone aware of a preferential stain for Methyl Violet or for oleic acid residues that would work on paper (cellulose and CaCO3 with traces of NaCl).
A colleague of mine is trying to decipher text printed with Methyl Violet pigment in an oleic acid carrier. In attempts to wash off obscuring blood stains, the paper was soaked in saline and then in petroleum ether. Needless to say, the ether took off most of the ink, too. It would be useful to amplify any residues with a preferential stain, but that is out of my area of expertise. Any suggestions on this would be appreciated.
I have a Polaron E5400 sputter coater that just sputtered out. It was coating very slowly so I put in a new gold/paladium target (the old one had burned through). When I went to sputter the thing glowed briefly & then just quit. The mAmp scale pegged on the upper end. I did the obvious things like check the fuses and they are fine, though I still might put in new ones just in case. Any thoughts as to what happened? Does anyone know who works on these things now? Or maybe some company that sells them could help. I will gratefully accept any help or information that y'all might send my way to help me repair my fizzled sputterer.
Model: Polaron E5400 Sputter Coater with a Polaron E5500 Film Thickness Monitor.
Crying & sighing but not sputtering,
Paula :-(
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I would like to obtain an adapter that fits onto a 4x5 camera (much like the Polaroid film packs do) AND that has a T-mount on the center that would allow me to attach a 35 mm SLR camera onto the adapter. Anyone know where to find such a device? Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Paula, If the circuitry inside the box is at all like that of the E5200 then there is a large, high value resistor on the output side of the HT transformer. This resistor in our E5200 has overheated and failed a few times until we replaced it a few years ago with a higher wattage resistor. This now works without trouble. The original problem is that the HT circuit is protected by a fuse, but the rating of the fuse is such that the resistor becomes the weakest link in the chain, with the resistor protecting the fuse.
} } My Fellow Board (bored?) Members, } } I have a Polaron E5400 sputter coater that just sputtered out. It } was coating very slowly so I put in a new gold/paladium target (the old one } had burned through). When I went to sputter the thing glowed briefly & } then just quit. The mAmp scale pegged on the upper end. I did the obvious } things like check the fuses and they are fine, though I still might put in } new ones just in case. } Any thoughts as to what happened? Does anyone know who works on } these things now? Or maybe some company that sells them could help. } I will gratefully accept any help or information that y'all might } send my way to help me repair my fizzled sputterer. } } } Model: Polaron E5400 Sputter Coater with a } Polaron E5500 Film Thickness Monitor. }
Prof Jan Coetzee Head: Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-362-5150 Pretoria 0002 Internet:janc-at-ccnet.up.ac.za South Africa http://www.up.ac.za/science/electron/emunit1.htm
I would guess that you have lost your transformer. It is not an uncommon problem and it is a reasonably easy job to replace it. Some transformers last for ever and some fail after a couple of years - it may just be luck.
Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
My experience of the Polarons (we have two) is that they are not very complex instruments that may be quite easily repaired by a competent electronics repairperson. The specialist skills (?) of an agent are not likely to be necessary. Jan Coetzee's experience is one example, we had a transformer fail, also not difficult to diagnose
Tony
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg KwaZulu Natal, South Africa
id AA01376; Fri, 27 Mar 98 07:33:39 CST Received: (from x400-at-localhost) by mta.mcdermott.com (8.7.1/8.7.1) id HAA28593; Fri, 27 Mar 1998 07:14:37 -0600 (CST) X-Authentication-Warning: mta.mcdermott.com: x400 set sender to Woody.N.White%650-at-wtgw.mcdermott.com using -f Received: by MCI; Fri, 27 Mar 1998 8:03:00 -0600
First, be careful! Sputter coaters use 1000 to 2000 volts at lethal currents.
A key bit of information is pegging of the current meter. I would need circuit schematics to be absolutely certain, but if conventional circuitry is employed.... This would not indicate a failed current limit resistor resistor unless shorted to case ground. Also, if the fuses were open or the transformer had failed NO current would result, not over current. The current meter should be in series between the high voltage source and the "load" (target). Over-current would typically result from a short / low resistance path between the meter and the target / bell jar.
With the power removed and the H.V. supply shorted with a jumper (for safety - also BEWARE of any capacitors which may hold a charge), disconnect the H.V. line (where exactly depends on circuit details) and measure the resistance. The value should be infinity between the line to the target and ground return. Find and remove the short circuit.
BTW, you might want to verify the over-current did not hurt the diodes also. The old cliche is that since semiconductors are so fast, they will usually blow to protect the fuse.
-------------------------
Woody White McDermott Technology, Inc woody.n.white-at-mcdermott.com http://www.mtiresearch.com
Home woody.white-at-worldnet.att.net http://www/geocities.com/capecanaveral/3722
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My Fellow Board (bored?) Members,
I have a Polaron E5400 sputter coater that just sputtered out. It was coating very slowly so I put in a new gold/paladium target (the old one had burned through). When I went to sputter the thing glowed briefly & then just quit. The mAmp scale pegged on the upper end. I did the obvious things like check the fuses and they are fine, though I still might put in new ones just in case. Any thoughts as to what happened? Does anyone know who works on these things now? Or maybe some company that sells them could help. I will gratefully accept any help or information that y'all might send my way to help me repair my fizzled sputterer.
Model: Polaron E5400 Sputter Coater with a Polaron E5500 Film Thickness Monitor.
Crying & sighing but not sputtering,
Paula :-(
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Over the year since I joined the list server there is an area of our "sport" that constantly hits me in the eye.
People keep talking about OPTICAL microscopes, what does this mean?
We do have light microscopy, transmission electron microscopy, scanning electron microscopy and the modern light scanning systems, are they not ALL optical microscopes some of which use ELECTRON OPTICS. Without optics in electron microscopes how would they work?
How did we get into this mess and do we let it continue?
That should stir a few minds!
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
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Grace, Just handle routinely like any oather biological sample. Incubating cell with resin beads has been done for many years. The resulting preps can be used for regular TEM or for ICC. Embed in either epoxy or in Spurr resin. Both should work. I believe LRW would also work although I haven't tried it. I did some years ago where we incubated the cell/bead prep with antibody and ferritin prior to fixation and embedding and got great immuno labeling....this was before colloidal gold was readily available.
Debby Sherman, Manager Microscopy Center in Agriculture Purdue University 765-494-6666
--------------------------------------
Help! I'll be receiving some resin beads with cell organelles (presumably) attached soon and I've never dealt with this sort of prep before. I would appreciate hearing from anyone out there with helpful information about the processing, handling etc. of these things. I've been off the listserver for a while so if it's already been discussed, please just answer privately. Many thanks. Grace
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I cannot take credit for this handy technique. This method was worked out and published by Dr. Ron Luftig in 1967 as part of his thesis work. The reference is as follows: Luftig, R. and R. Haselkorn. (1967) Morphology of a Virus of Blue-green Algae and Properties of Its Deoxyribonucleic Acid. J. of Virology Vol. 1, p. 344-361. Dr. Luftig calibrated beef liver catalase crystals (available from Sigma and other sources) using copper phthalocyanin crystals and Fullam replica gratings. He determined that the line spacing of the catalase crystals was 91 +/- 3 angstroms by UA and PTA staining. The exact technique was to place a droplet of virus suspension on a formvar coated grid. Then invert the grid onto a droplet of 2% UA for 90 sec. Next float the grid on a solution of 0.08ml catalase crystal suspension in 5ml of 2% UA for 2 min. Draw off excess stain, etc. with filter paper and dry. He prefered this method to mixing the virus and catalase together prior to staining. This method yields virus sitting on and adjacent to the catalase crystals and gives a very easy method of determining virus size. One suggestion is to check the catalase suspension when it is received. We have gotten preps where the crystals have broken down. Catalase also is wonderful for more accurate calibration of TEM's at higher magnifications than can be done with replica gratings.
Debby Sherman, Manager Microscopy Center in Agriculture Phone: 765-494-6666 Purdue University FAX: 765-494-5896 W. Lafayette, IN 47907
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Would you be interested in writing a short article or technical tip on using catalyse crystals as a standard for EM? And in making the standard, if that is part of your procedure. Our readers would find this information useful.
If you don't get Microscopy Today, it is a trade journal sent free to microscopists in universities, industry, hospitals and government labs. If you would like a subcription, please send me your preferred mailing address.
Thank you for your attention.
Phil
} I do always recommend internal standards with specimens whenever possible. } One of } the classics in biological work is catalase with a very well } defined crystalline } spacing combined with preps of virus and small protein molecules. Accurate } measurements of particles adjacent to or resting over these lattices eliminate } problems with hysterisis and all other potential situations which may effect } magnification. } Debby Sherman
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 5037 Station A Champaign, IL 61825-5037 oshel-at-shout.net or poshel-at-hotmail.com
Hello everybody, I would appreciate to hear from anybody who has an experience (positive or negative)labelling paraffin sections with antibodies to matrix metallproteinases (MMP's) and their inhibitors (TIMP's). I have tried some antibodies but they seem to expire very quickly. Thanks for any info.
Sarka Lhotak
Hamilton Regional Cancer Centre McMaster University Hamilton, Ontario, Canada lhotaks-at-mcmaster.ca
"Microscopy and Imaging Resources on the WWW" http://www.pharmacy.arizona.edu/exp_path.html
Microscopy and Imaging has recently been updated (3/15/98) and is sporting a new interface. To find this site please go to the URL above and look for the link to "Microscopy and Imaging" in the middle of the page. I would offer a more direct link to the page, but in the past the URLs have been so long that many email programs cause them to wrap to the next line and my webmaster hates it when visitors get "404 page not found" messages.
The following pages are included in the "Microscopy and Imaging Resources on the WWW" site; Histology, Confocal Microscopy, Electron Microscopy, Digital Imaging, Specialty Microscopic Techniques and a list of Free Publications of Interest to Microscopists.
Microscopy and Imaging was initially my way of keeping track of the various sites on the web that feature materials that would help local grad students, staff and faculty who were unfamiliar with microscopy & imaging to learn about the different techniques. I also wanted them to have access to some of the terrific technical reference materials out there. The WWW site has grown (25,000 visits in the last year) to become as much a resource to the scientific community (consistent with our NIH funding) as it is a point of reference for local users.
Because Microscopy and Imaging is primarily a meta-list of links to other places, I want to take this opportunity to acknowledge (in a far too general way) that there are many terrific individuals, labs, universities and companies out there who provide accessable and well written educational or reference material related to microscopy and imaging.
Submissions and suggestions are encouraged (see comment below):
It is not consistent with the purpose of this site to list every microscopy and imaging related company, lab or society on these pages. There are MANY excellent sites that do this and I have tried to provide links to several of those sites. I am, however, always looking for new sites that do provide well done educational or reference materials related to microscopy and imaging. (A note to commercial vendors: it is always tricky to evaluate commercial sites, I tend to link to sites that have strong content and are low-key in making reference to particular products.)
Yours, Doug Cromey ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
Dear Paula, We have had open houses at our university several times in the past, and we find the EM lab to be a very popular exhibit. Just keep the instruments running and allow people to see samples live in them. An insect in the SEM is very popular, also flower pollen and cross-sectioned wood. The TEM is impressive by itself, but if they can see a sample live or on a TV monitor it is even better. You wrote: } Websters, } } I'm looking for a copy of the February 23, 1998 TIME magazine } (vol.151, no.7). It's the one with the cover of a negative stain of the } Hong Kong chicken/human flu virus & a scientist in a "bunny suit". We're } going to have our lab available during campus open house & thought that the } article, picture, etc. was a good way to show the general population how } TEM's help us in the real world. } If anyone knows of a source or has one hanging around that they no } longer want, please let me know. } Also, if there is a location where we can find pictures of Ebola or } any other viruses please let me know. We think that most people know what } viruses are so we'd show them instead of trying to explain what the inside } parts of a cell are. } } We've never had open house before, so if y'all have any cool } suggestions for displays send 'em my way. } } Thanks Kiddos! } } } } Paula :-) } } Paula Sicurello Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
I am forwarding the following inquiry for a colleague here. If anyone has any suggestions, would they please send them to Jerry at his e-mail address. TIA.
Gill Bond Dept Materials & Met. Eng. New Mexico Tech
I am trying to detect a protein between inorganic lamellae (CaCO3) using SEM. I am unsure if I can get enough metal with a standard dye to show up using EDS, and it was brought to my attnetion that I could use dot mapping as a superior tool. What I am asking, is if anyone knows if a TEM organic dye would have enough metal to be detected. Any suggestions??
Captain Semantics, here, on the subject of 'optical' microscopy.
} From Websters 7th new collegiate dictionary, optics is "...a science that deals with light, its genesis and propagation,...and other phenomena closely associated with it." The two definitions of 'optic' refer to the eye, lens, etc. and eventually refer back to the science, 'optics'. The word 'optical' similarly traces back to the science of 'optics', so I think optical microscopy properly refers to the use of visible light to image structure. It seems to me that we have chosen to view our speeding electrons as very high energy waves (which I think is legitimate), hence the use of photon terms, even though the lenses work on a particle-moving-in-a-field basis instead of the propagation rate (RI) basis usually used to understand optical lenses. At least there is some physical basis for this term usage since both of the 'optics' systems work to focus the probing particle/wave. (I suspect a good theoretician could prove the equivalence, but that is beyond me today!) By counter example, consider the NMR community which refers to the signal generated by the precessing nuclear moments as a 'free induction decay' or FID. This term is (at least in my opinion) a hold-over from the early definitive NMR experiments where the actual magnetization DECAY was of interest. In fact the FID is the envelope that grossly defines the overall drop in transverse magnetization. What we actually use, but casually call the FID, is really the beat frequency pattern due to nuclei precessing with varying frequencies. The pattern decays in time due to relaxation and other phenomena such as magnetic field inhomogeneity, but the decay is usually not appreciated by those attempting high-resolution NMR for compound identification! I tried to convince my thesis adviser that we should call it the free induction signal since it is the signal induced in the detection coil from freely-precessing nuclei, but he just raised his eyebrows. It's tough to change 50 years of literature...
Bill Heeschen Optical AND Electron Microscopist ;-) The Dow Chemical Company
MICROSCOPICAL SOCIETY OF CANADA MAY 27th-29th 1998, MONTREAL, QUEBEC
INVITED SPEAKERS
Dr. David Joy, University of Tennessee, Knoxville =22Scanning electron microscopy=22
Dr. Eric Lifshin, GE Corporate R&D, Schenectady, NY =22X-Ray microanalysis=22
Dr. Pierre Hovington, IREQ, Qu=E9bec, Canada =22Scanning electron microscopy=22
Dr. Gianluigi Botton, CANMET, Ottawa, Ontario, Canada =22Electron energy loss spectrometry (EELS) =22
Dr. Sophie Boisvert, Noranda, Montr=E9al, Qu=E9bec =22Material sciences=22=20
Dr. Wah Chiu, Baylor College of Medecine, Houston Texas=20 =22Electron Cryomicroscopy for Macromolecular Assemblies below 10 A.=22
Dr. George Harauz, Guelph University, Ontario Canada =22Structure-function relationships of myelinic proteins: basic protein, = lipophilin, and surfactant protein A=22.
Dr. Dale Laird, University of Western Ontario , Ontario, Canada =22Imaging of connexin 43-GFP chimeras in living cells in real time.=22
Dr. Louise Brisson, Universit=E9 Laval, Qu=E9bec, Canada =22Plant Apoptosis, a distinct case of programmed cell death.=22
Dr. E. Sanders, University of Edmonton, Alberta, Canada =22Apoptosis During Early Embryonic Development=22
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According to definitions in my dictionary, "optic," "optics," and "optical" refer to the visible region of the electromagnetic spectrum--i.e. that part that can be detected with the eye. Therefore, "optical microscope" is probably a better term than "light microscope," after all our lab has an infrared microscope which utilized infrared light. "Light" sometimes refers only to the visible part of the spectrum, but also can refer to infrared, ultraviolet, and even x-rays!
I think "electron optics" is simply a term coined for the parts of an electron microscope that mimic the corresponding items on an optical microscope that perform functions like focusing, etc. It is a borrowed term and therefore may not be entirely accurate in the purest sense of the words.
In short, it may come down to semantics. I prefer the use of the term "optical microscope" because I think it more precisely defines the item in question. If I refer to the IR microscope, I call it the "IR microscope" (even though it also uses visible light for alignment of the sample!). I also may use the term "electron optics" from time to time without feeling guilty.
Jim Passmore Cryovac North America james.passmore-at-grace.com (for a few more days until Cryovac becomes part of Sealed Air Corp.)
---------- } From: PROTRAIN } To: Microscopy } Subject: Optical Microscopes } Date: Friday, March 27, 1998 5:22AM }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi- I'm almost embarassed to ask this question, but perhaps someone has the simple solution that has eluded me. I cut a lot of thick sections of large block faces (epoxy resin, 5mm X 5 mm), which are counterstained with toluidine blue/safranin. Off and on, I have a problem with stain drying lines running through the section. I can usually get rid of this by putting a drop of water on the section and placing the slide back on the hotplate for a minute or two. For the number of samples that I cut, this is time consuming. Does anyone have a solution for this problem? I use pre-cleaned slides straight from the box and rinse them with di water or 95% EtOH before I collect the sections. The slides are left on the hotplate from 4-24 hours (to ensure that the entire section has adhered to the slide) and then stained. I would appreciate any suggestions.
Thank you and have a sunny weekend,
Sandy Perkins
Laboratory for Neurotoxicity Studies VMRCVM VA Tech
{ {People keep talking about OPTICAL microscopes, what does this mean?} }
I wonder if the useage varies in the USA from Europe. I live here in the USA but am British and have always used the term Light Microscope (rather than Optical).
"Optical" suggests "visual" and originally was connected with the eyes and therefore with light (I assume?). Since the concept of creating images using energy other than light is recent, I would suggest that the use of the word "optics" in "electron optics" is probably inaccurate but should now be acceptable by virtue of its widespread use. Does anyone agree?
The term "optical microscope" is confusing though by any standards since, as you point out, an electron microscope uses a system known as "electron optics". So I would like to start an association known as "The Association for the Preservation of Pure Queen's English as Applied to Modern Science" :-) A required qualification for membership would be willingness to use the term "light microscope" when referring to those good old instruments with glass lenses :-}
I hope we can quickly resolve this matter and that it is not magnified out of proportion. Let's be clear on our objective.
Most reasonable people understand the following terms:
light microscope electron microscope (SEM or TEM) confocal microscope scanned probe microscope (more specific as needed) IR microscope x-ray microscope
"Optical microscopes", while technically correct, is confusing to a number of people.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I would certainly agree with John, and with the previous message by Ted Dunn. I have always preferred the term "light microscopy," and have considered "optical microscopy" to be equivocal and undesirable in our modern world, where electron optics is firmly established (and not a particularly new term -- solid work on the subject was being done in the 1920s). One certainly would not consider the magnificent three-volume work of Hawkes and Kasper, "Principles of Electron Optics," to be a misnomer. A dictionary is only worth buying if it reflects modern usage.
A. Kent Christensen Department of Anatomy and Cell Biology Medical Sciences II Building University of Michigan Medical School Ann Arbor, MI 48109-0616 akc-at-umich.edu Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
----------------------------------
On Fri, 27 Mar 1998, John J. Bozzola wrote:
} Most reasonable people understand the following terms: } } light microscope } electron microscope (SEM or TEM) } confocal microscope } scanned probe microscope (more specific as needed) } IR microscope } x-ray microscope } } "Optical microscopes", while technically correct, is confusing to a number } of people. } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ####################################################################
Hoping once again to tap into the collective wisdom of the listserver members. What would account for the following symptoms in an environmental SEM (Hitachi S3200N, to be precise):
---Inability to get rid of lateral motion during objective aperture centering (i.e., when the current is being pulsed to the objective lens, the image continues to rock back and forth despite use of the centering controls on the aperture). Normally the brightest screen image also closely corresponds with aperture center, but not even close in this case.
---Extreme difficulty in stigmating. Also at very low magnification there is a central brigher area on the screen with darker "shadows" at the sides. These shadows move and rotate when the stigmator adjustments are used.
---Extremely poor resolution, even at mags as low as 1000x.
The filament was changed recently by a Hitachi technician. Filament centering has been checked and rechecked. The gun is clean. The objective aperture strip was just cleaned by baking in a vacuum evaporator. Complete column alignment has been done: the filament image is centered, gun tilt and horizontal have been repeatedly checked, both manually and using automatic gun alignment. Coarse and fine stigmation has been repeatedly performed.
Except for a dirty aperture strip, focus and aperture centering was fine before the filament change, with high quality images past 20,000x. The problem appears to be related to the filament change, although it has also occured in the past, when it seemed not to be related.
I am guessing that there may be debris on a fixed aperture or a dislodged or uncentered fixed aperture, but I am looking for any suggestions. Thanks in advance for help with this puzzler.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
I have been staining some LRWhite(medium) sections and I am getting very low contrast with 30min. 3%UA and 10 min. Lead Citrate. I have tried reducing the wash times and the KV to 60KV without any luck. Any ideas and/or advice?
Barbara Plowman University of the Pacific School of Dentistry 2155 Webster San Francisco, CA 94598 email:Bplowman-at-uop.edu
Gillian Bond wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am forwarding the following inquiry for a colleague here. If anyone has } any suggestions, would they please send them to Jerry at his e-mail } address. TIA. } } Gill Bond } Dept Materials & Met. Eng. } New Mexico Tech } } I am trying to detect a protein between inorganic lamellae (CaCO3) } using SEM. I am unsure if I can get enough metal with a standard dye to } show up using EDS, and it was brought to my attnetion that I could use dot } mapping as a superior tool. What I am asking, is if anyone knows if a TEM } organic dye would have enough metal to be detected. Any suggestions?? } } Jerry Egeland } eggman-at-nmt.edu
Jerry
If you have line profile capabilities, your sensitivity will be much greater. Following a line that goes up and down is much easier for the mind to work with than dot densities. Even gray levels are easier for the mind to discern than dot densities so if you have to stick with dot mapping due to lack of a rate-meter, take your dot map and then defocus the image if your concentrations don't appear to be high enough to discriminate.
Ken Converse owner Quality Images third party SEM service
1998 Spring Joint Meeting The Texas Society for Electron Microscopy and The Oklahoma Microscopy Society April 2,3, and 4, 1998 Lake Texoma Resort
"Back to the Future: Improving Your Image".
As of Friday March 27, 1998, the most up-to-date program schedule (including definitive times, room numbers, etc.) will be listed on the Oklahoma Microscopy Website. General info is listed there now. I do have the program on file if any one would like it ahead of time. If you have any questions, please do not hesitate to contact me.
website: http://www.ou.edu/research/electron/oms/
Sincerely,
Ginger
Ginger Baker EM Lab Manager OMS Secretary/Treasurer Research Dept., OCOM 1111 W. 17th St. Tulsa, OK 74107 Phone: (918) 561-8232 FAX: (918) 699-8629 http://osu.com.okstate.edu/dept/research/content/gbaker.htm lizard-at-osucom-fs02.ocom.okstate.edu
I'm wondering that whether there is any microscopic research concerning diffusion rate of any dyes in starch. Also, I'd like to know how I can search for these kinds of materials and the images obtained from this method. Could you please give me some information? Thank you very much.
We are looking for a donation of a Balzers freeze fracture machine, preferably 300 series, that we could use for double-layer coating (platinum/carbon) for low-temperature SEM. A conventional machine would do. This would be for the University of California, Berkeley. Please reply if you know of anyone with one of these machines; any leads would help.
Thanks, Jacob Bastacky
Jacob Bastacky, MD Room 116 Donner Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Telephone: 510.486.4606 Berkeley, California 94720 FAX: 510.845.8031
Wei Chen wrote: ================================================= Regularly TEM grids are 3.05 mm in dia. and about 6 um in thickness. Does anyone have the info on who is making larger and thicker grids? Thanks a lot in advance. =================================================
I will assume that you are talking about the "typical" grids made by electrodeposition techniques (as opposed to grids made by the etching of a foil which are also a lot more expensive but they do tend to be thicker).
For the "normal" 3.05 mm grids, there are no manufacturing or technical reasons why a grid can not be made thicker. But the reasons why they are not routinely made thicker is that when doing tilting experiments, the thicker the grid, then the smaller the percent open area. Generally speaking, the grids are made to a thickness that they are "self supporting" but not beyond that point for the reason just given.
We can provide grids of just about any thickness one might want. However, the "catch" is that there is some minimum number that one would have to make (for decent economics) and usually when that number is heard, some of the original enthusiasm for that thicker grid gets lost.
If you want grids larger than 3.05 mm, the best (e.g. most economical) approach is to work with grid mesh (see our website at address given below), cutting out the diameter that you do want. The grid mesh comes in pieces as large as 6" square. This is the same mesh used for making distortion adjustments in an SEM. You can select from Cu, Ni, or Au. Special art work could always be made to produce a specifically larger diameter and thicker grid from scratch but you are starting to talk about big bucks.
Contact me off line for any further discussion about how we could help you find what you need.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I am a microprobe analyst at RJ Lee Group In Pennsylvania. I am in desperate need for low voltage DC 1670:1 crystal flipping motors. I am currently engaged in obtaining line profiles through a metal reaction zone. The analysis calls for 60 points. The problem is, however, that one of our detectors is not working correctly (Causes Sandia Task8 to crash when it trys to change the angle theta of the detector (any ideas on this one? Seems like it thinks the detector has reached a limit, but it hasn't). Therefore, we are limited to obtaining five elements on the same spectrometer using three of the four crystals on that spectrometer. We have ruined two motors so far (they make a horrible squealing noise and don't seem to seat properly). Any help would be greatly appreciated. Thanks
I have been contacted by someone who is working on an MA in Design and Media Arts who has been using an old SEM in a hospital for some of their work. They would like access to a newer SEM with better resolution, if possible. London area is preferred, but anywhere in the UK would be OK. Is anyone out there willing to help out? If so, please e-mail me, and I will forward the messages.
Mahalo to you all, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
You have no doubt already been innudated with suggestions for insects, mites and microbes, but the greatest hurdle to overcome in explaining EM is the range of magnifications available. I would suggest that you start out with a macro view of something like a running mechanical watch movement. In fast scan or TV rate mode, it can provide a good starting point along with an interesting perspective. Follow it up with increased magnifications on the metallic surfaces of the gears or frame and you can provide the magnification steps that are usually missing.
After that you can change samples to an insect and pick-up the magnification relationship by zooming in on the multiple eye lenses or body hair. If possible, use light microscopy images to explain the capabilities of light vs. electron microscopy.
We often take for granted the dimensions of the work we do. To the layman, however, these dimensions mean nothing. Whatever you can do to accentuate the relationship between everyday dimensions and the esoteric realm of EM will result in greater understanding.
} Dear Paula, } We have had open houses at our university several times in the past, } and we find the EM lab to be a very popular exhibit. Just keep the } instruments running and allow people to see samples live in them. An } insect in the SEM is very popular, also flower pollen and } cross-sectioned wood. The TEM is impressive by itself, but if they } can see a sample live or on a TV monitor it is even better. You } wrote: } Websters, } } I'm looking for a copy of the February } 23, 1998 TIME magazine } (vol.151, no.7). It's the one with the } cover of a negative stain of the } Hong Kong chicken/human flu virus } & a scientist in a "bunny suit". We're } going to have our lab } available during campus open house & thought that the } article, } picture, etc. was a good way to show the general population how } } TEM's help us in the real world. } If anyone knows of a } source or has one hanging around that they no } longer want, please } let me know. } Also, if there is a location where we can find } pictures of Ebola or } any other viruses please let me know. We } think that most people know what } viruses are so we'd show them } instead of trying to explain what the inside } parts of a cell are. } } } We've never had open house before, so if y'all have any } cool } suggestions for displays send 'em my way. } } Thanks Kiddos! } } } } } Paula :-) } } Paula Sicurello Regards, Mary Mary Mager Electron } Microscopist Metals and Materials Engineering University of British } Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: } 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Dear all We wish to use our PC running Windows 3.1x to print out a list of all the file directories and image files that we keep on MO disk and CD. We can do this on a Macintosh platform, and to a limited extent with DOS, but want to use Windows to order and sort our data.
Please reply to me, and I'll summarise any sucessful method on the listserver in due course. Thank for your help & interest.
I am an undergraduate at the University of Rochester and I am trying to look at neurons under the SEM. I already have the brain sample but I have no idea how to separate the neurons from the gray matter and fix them for SEM viewing. Please help. Thank you in advance.
At 11:30 AM 3/30/98, you wrote: } } Dear all } We wish to use our PC running Windows 3.1x to print out a list of all } the file directories and image files that we keep on MO disk and CD. } We can do this on a Macintosh platform, and to a limited extent } with DOS, but want to use Windows to order and sort our data. } } Please reply to me, and I'll summarise any sucessful method on the } listserver in due course. Thank for your help & interest. } } Jeremy.Sanderson-at-path.ox.ac.uk
To produce a file listing all the image files in a directory and all subdirectories in Windows 3.1 you can use something like the following command at the DOS prompt to create a text file containing the file names, dreation dates, and sizes.
dir *.gif /s } images.txt
dir is the directory command
*.gif specifies all files with a gif extention
/s specifies that the dir command is to include all subdirectories
} images.txt redirects the output of the dir command to the file images.txt.
The output of this command looks like:
********************************************************** Volume in drive C is MS-DOS_622 Volume Serial Number is 1234-1234
You can then import the text file into a spreadsheet program, cleanup any extraneous information and sort the file list anyway you want. You can also add data fields.
Steve Chapman is right: there is a small lack of precision regarding the terms LIGHT microscope and OPTICAL microscope. I used the term LIGHT-OPTICAL microscope for some time before Barry Carter suggested VISIBLE LIGHT microscope. Barry's suggestion removes all uncertainty and we can all now get some sleep nights. :-)
To all who very kindly replied to my SEM question, thanks!!! The trouble turned out to be in the final polepiece aperture, which came out in three pieces when I checked it. Luckily, it fell into my hand, rather than disappearing forever under the baseplate of the chamber. After cleaning it, reassembling it, and reinserting it, we had a scope again.
The responses I received were very interesting and helpful. Some were quite extensive and must have taken some real time to put together. I'm making a file of them for future reference in troubleshooting, and I would be happy to forward a summary to the list, if desired. Thanks again to everyone who helped.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
To the Light Microscopists, Is it possible for one week old paraformaldehyde (originally in argon sealed ampules)-EM grade- to actually permeabilize membranes? (by some glycol formation). All comments are welcome.
Does anyone have experience growing tissue culture cells on gold grids with support films? We want to do IEM of filaments after digestion of the cytoplasmic membrane,
Articles describing this technique mention specific kinds of reagents, grids, etc. Are there better brands, methods? We'd appreciate any tips to avoid a lot of trial and error.
Source of gold grids? (supplier) Type of support film? Formvar/collodion) Carbon coat? (heavy/light) Method of sterilization? (UV/gas) Type of cell membrane solvent? (detergent/organic) Other hints?
Thanks, Sara
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Dear Steve, } } Over the year since I joined the list server there is an area of our } "sport" that constantly hits me in the eye. } } People keep talking about OPTICAL microscopes, what does this mean? } } We do have light microscopy, transmission electron microscopy, scanning } electron microscopy and the modern light scanning systems, are they not ALL } optical microscopes some of which use ELECTRON OPTICS. Without optics in } electron microscopes how would they work? } Remember, there are also scanned probe microscopes. Some of these (SEM is arguable) do not need optics. Also, IMHO, calling the beam focus- sing system of an electron microscope "electron optics" is an analogy with light optics, not an identical process, so retaining the nomenclature "op- tical microscope" to mean one with light-optic lenses is not a "mess".
} How did we get into this mess and do we let it continue? } } That should stir a few minds! } I prefer mine shaken. Yours, Bill Tivol
Sir, dear Brett, If you don't get a commercial mold for this purpose I eventually can = send you a self made silicone rubber embedding mold which perhaps fits = your needs (please specify diameter or special shape of the mold needed; = I assume you want to embed into resin?, paraffin??, cow's (bull's) = eye(s) or fish's eye(s)?? or.... If your needs is a metal embedding mold, sorry, I don't have.
Best regards Wolfgang
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
I'm looking for metal embedding molds that are deep enough (15-20mm) to embed whole eyes. Can someone please post the name(s) of the = supplier(s). Thanks.
Dear Randy, My guess is you are right, there is some small bit of something in the column. You should get the column cleaned, as there are four or five fixed apertures down the column and at least one below the final aperture. The other thing to check is the alignment of the first and second condensor lenses. Also make sure you are not one aperture hole out, the controls have enough adjustment to get you to the next hole in the strip. Start with no aperture and count them in, adjusting brightness and centering each as you reach it. The stigmation cannot be corrected until the aperture is straight (doesn't move when you wobble the lens current). Centre as many controls: stigmators, etc. as possible before aligning. I don't have a low-vac SEM, just an old S-570, but that is what I've found. Getting the aperture straight may solve all your problems at once. You wrote: } Hi, } } Hoping once again to tap into the collective wisdom of the listserver } members. What would account for the following symptoms in an environmental } SEM (Hitachi S3200N, to be precise): } } ---Inability to get rid of lateral motion during objective aperture } centering (i.e., when the current is being pulsed to the objective lens, } the image continues to rock back and forth despite use of the centering } controls on the aperture). Normally the brightest screen image also } closely corresponds with aperture center, but not even close in this case. } } ---Extreme difficulty in stigmating. Also at very low magnification there } is a central brigher area on the screen with darker "shadows" at the sides. } These shadows move and rotate when the stigmator adjustments are used. } } ---Extremely poor resolution, even at mags as low as 1000x. } } The filament was changed recently by a Hitachi technician. Filament } centering has been checked and rechecked. The gun is clean. The objective } aperture strip was just cleaned by baking in a vacuum evaporator. Complete } column alignment has been done: the filament image is centered, gun tilt } and horizontal have been repeatedly checked, both manually and using } automatic gun alignment. Coarse and fine stigmation has been repeatedly } performed. } } Except for a dirty aperture strip, focus and aperture centering was fine } before the filament change, with high quality images past 20,000x. The } problem appears to be related to the filament change, although it has also } occured in the past, when it seemed not to be related. } } I am guessing that there may be debris on a fixed aperture or a dislodged } or uncentered fixed aperture, but I am looking for any suggestions. Thanks } in advance for help with this puzzler. } } Randy } } } Randy Tindall } Electron Microscope Laboratory } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } } rtindell-at-nmsu (work) } nrtindall-at-zianet.com (home) } Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Thanks for your message the other day. I am now trying to work out some of the specifics of the buffer. What osmolarity did you use? In a paper on IVF they suggest 285-295 Osm, yet I can't use this buffer because it has albumin (bad for glutaraldehyde).
Is there a reference that may have this information? I have exhausted several search pathways with no progress.
Thanks, Todd Rippy University of Oklahoma, Department of Zoology
} } We do have light microscopy, transmission electron microscopy, scanning } } electron microscopy and the modern light scanning systems, are they not ALL } } optical microscopes some of which use ELECTRON OPTICS. Without optics in } } electron microscopes how would they work? } } } Remember, there are also scanned probe microscopes. Some of these } (SEM is arguable) do not need optics. Also, IMHO, calling the beam focus- } sing system of an electron microscope "electron optics" is an analogy with } light optics, not an identical process, so retaining the nomenclature "op- } tical microscope" to mean one with light-optic lenses is not a "mess". } } } How did we get into this mess and do we let it continue? } } } } That should stir a few minds! } } } I prefer mine shaken. } Yours, } Bill Tivol
Gee Bill,
I am not alone. It grates on me seeing that analogy turn into a definition. While I see the point of using the term LIGHT MICROSCOPE, for me electron optics are not optics. But then I am an old sh.t and one in the definite minority who still likes to play with light, and could give a whoop agout an electron. So I guess I wil just have to mumble in my single malt and let majority rule.
Shalom from Jerusalem, Azriel +++++++++++++++++++++++++++++++++++++++++++++++++++++ Major Azriel Gorski, Head Fibers and Polymers Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem, ISRAEL
azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739
What lies behind us and what lies before us are tiny matters compared to what lies within us. Ralph Waldo Emerson ++++++++++++++++++++++++++++++++++++++++++++++++++++
I am sorry to hear you have had a problem with an E5400 sputter coater. Although the E5400 has not been manufactured for nearly 10 years we do still carry the spare parts to support this instrument. Our local agent, Energy Beam Sciences Inc. (ebs-at-ebscience.com telephone: 413 786 9322)) have been our US agent since 1991 and would be pleased to offer spare parts and support for Polaron instruments.
Since the first Polaron coaters were introduced into the market back in the early 1970s several thousand sputter coaters (and a similar number of critical point dryers) have been sold into EM laboratories around the world. We pride ourselves in efforts to support instruments currently in the field - many now well over twenty years old.
Some customers do have difficulties locating us due to changes in company name and local agents during the middle part of our history. For the record the original company; Polaron Equipment Ltd, became part of Bio-Rad Microscience (UK) in the mid-1980s. At that time the Polaron name was dropped from the instruments. In 1987 Bio-Rad Microscience purchased another UK manufacture of EM preparation equipment; Emscope Laboratories Ltd. Finally in 1991 the Polaron range was acquired by VG Microtech (then owned by Fisons, now by Thermo Instruments) and relocated to our present factory in Sussex. At that time the Polaron name was brought back is now proudly displayed on all of our products.
To illustrate the fact that the Polaron brand is very much alive, we have recently launched a new Cryo-SEM system and will shortly be introducing a new sputter coater to the Polaron range.
Best regards M.J.Wombwell Polaron range Business Manager http://www.vgmicrotech.com/polaron-range
Dear listees, I want to do some 3 dimensional reconstruction of a particular type of fibroblast in normal and pathological states, but I really am at a loss as to the methodology. I do not have access to a program to do this, so I need some sort of 'old-fashioned' but reliable method. Any help will be gratefully receieved (as will any cheques, real-estate, cars, post-doc offers etc. etc.). Thanks in advance, Yours 2 dimensionally, Alex
...................................................................... ....................................... Alex Black Department of Anatomy National University of Ireland Galway
Dear Robert, as I informed you in a previously sent information
(your Q sent off the MSAListserver dated 18th of March, 1998 :=20 } Since you are familiar with embedding samples, do you know of an embedding resin that will not shrink upon curing but still has a low viscosity?
Dr. Robert Dickson Finnish Pulp and Paper Research Institute {
My reply then was: } 18th of March, 1998: 11.05 a.m.
Dear Robert, thank you for your confidence in my knowledge. } From my experience I know to some extent my embeddings with the = Epon-recipe (now Glycidether 100/SERVA as EPON 812-substitute) used in = my Lab since 1982/83 would have about 3-5 % at the maximum. = Unfortunately I haven't done any calibration and/or measuring methods on = that. It's really estimated....(based on correct LM-measurements of = leading structures from tissue samples (like diameters of bacteria in = LM-semithin sections, compared to the diameters measured on correlative = TEM-ultrathin sections, compared to references from literature, or like = diameters of erythrocytes etc...). I assume you would like to embed paper or paper-like stuff and need a = low viscous embedding medium in order to measure by TEM, e.g., = length/diameters of cellulose/paper fibers. I regret: I don't have any = experience on such matter but I would be interested in hearing about = "how to do processing of paper samples".
!!!!!----- In my experience, shrinkage of embeddings by polymerization is not THE = point: I would guess specimen processing steps before infiltration of = the paper stuff would be. So I would need some informations more on = "what purpose, what was used/done until now, problems", etc. to be of = any help with respect to your question. Generally I think the = epoxy-resin class does have the lowest shrinkage factor as compared to = methacrylates.... but I would have to go into special literature for = that. If there is anything you think I could be of help, please let me = know and don't hesitate to contact me by e-mail..... !!!!!---------
For the moment, best regards
Wolfgang End of my response 18th March 1998 ------------------------------------------------------------------
So, also taking into former considerations of other list members on that = subject (confer: Tobias Baskin, 18th March; Caroline Schooley, 20th of March),=20 I really think your problem is *not shrinkage* of a particular resin but = instead -having chosen a particular resin type (I/we don't know at all) = establishing right standard procedures for dehydration, infiltrating = embedding as well as appropriate polymerization steps for *your = material* as well as that *particular chosen resin type*. I remember those days when being educated and learning during my = universitarian EM-studies about embedding that the outcome of "simply do = embedding" (in terms of sectioning quality, reliable results, etc.) = would depend on the steps DEHYDRATION (or any other pretreatment of = specimens), correct and appropriate infiltration steps (including the = right "intermedium", its evaporation), correct and appropriate = polymerization steps (delicate specimens need other polymerization = schedules than less delicate ones).=20
I think, your specimens (cellulose fibres) are more delicate than = optimally fixed, dehydrated and embedded/polymerized -say-normal animal = or human liver specimens, . Insofar I think that without exactly knowing your recent specimen = preparation schedule/steps one could not follow the empirical route of = trouble shooting. The only suggestion I could make according to the informations you told = us would be: if you polymerize only 48 h at 60 degr. C (what kind of Epoxy resin??, = which mixture??) I think this would be the worst you can do with such a = delicate specimen like cellulose fibres:=20 I read the comments of the original LUFT-articles on epoxy resin = embedding and the proposal(s) for slow, elongated epoxy resin = polymerization for delicate specimens carefully (37, 45, 60 degr. C = about 24 hrs each for better [section] quality of polymerized epox resin = blocks), did you??
An example?: I have tested now the LADD LX 112 Resin (an EPON 812 = subsitute) I got several weeks before. As usual, first of all one tests = a new resin under the conditions you had routinely had/done before = (despite affirmations "this or that particular resin/its properties = is/are like the ORIGINAL".....).=20 Then if you want, you can see the differences:=20
a "Rapid POLYMERIZATION of LX 112"=20 (i.e. without prepolymerization steps at lower temperatures but 1 h at = 90 degr. C)
yields other polymerization properties of resin blocks compared to a=20
"Rapid POLYMERIZATION of EPON 812, or another subsitute, GLYCIDETHER = 100"( also 1 h at 90 degr. C),=20
and, additionally, as this was my personal experience yesterday evening: = the properties of LX 112 *during* polymerization (i.e. viscosity changes = before polymerizing) are quite different than EPON 812 or the substitute = Glycidether 100 (given all other components of the resin mixture were = added as usual). The trimming/section quality has to be assessed then, you have to check = and -if needed- to adjust section speed, cutting angles, etc.
The problems of wrinkling sections, or -maybe as in your case- partially = loosing specimen material on sectioning (either semithin or ultrathin) = is widespread and sometimes are hardly to overcome.=20 BUT: trouble-shooting for this would be easier knowing what was done = before with the specimen. Maybe there would be a chance to overcome your = still existing problem.
best wishes regards,
Wolfgang
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
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I am embedding cellulose fibers in an Epoxy resin mixture. The mixture is cured for 48 hours at 60C. It appears to shrink and put the sample under compressive forces because after cutting, the slices contains waves and want to expand. The problem I am having is that the cellulose will not allow the resin to relax after cutting, thus the resin tears the cellulose fibers apart. =20
Does anyone have any suggestions for other low viscosity resins that will not shrink upon curing or general ideas for what I working with?
We're trying to make a deposition of 2 micron gold particles on a silica substrate. We cannot use an evaporator because we don't want a thin film: we want to deposit the particles directly on the substrate, keeping them isolated and without forming aggregates. At first we dispersed the powder directly on the silica, but SEM analysis showed they formed aggregates. Then we've tried making a suspension on etanol and then putting a droplet of the solution directly on the substrate. But still it formed aggregates.
Does anyone know of a simple method to make such a deposit (without forming aggregates) ?
Thanks in advance.
Isabel Nogueira Instituto Superior T=E9cnico Departamento de Engenharia de Materiais Av. Rovisco Pais 1096 Codex Tel.: 351 - 1 - 8418124/0 =46ax.: 351 - 1 - 8418120 E-mail: isabeln-at-alfa.ist.utl.pt
Every other person on the planet knows what a "light microscope" is so I will go my merry way and be ignorantly blissful ... and solving real industrial problems with whatever this thing is on my bench.
On to real issues!!!
Jim Harper Amoco
P.S. Prefer to call it a Polarized Light Microscope! How about Super Electric Microscope (SEM) for that big hunk of steel taking up the rest of my lab space?
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See very end for my addition...
(Much snipped for sake of bandwidth)
} } We do have light microscopy, transmission electron microscopy, scanning } } electron microscopy and the modern light scanning systems, are they not ALL } } optical microscopes some of which use ELECTRON OPTICS. Without optics in } } electron microscopes how would they work? } } } Remember, there are also scanned probe microscopes. Some of these } (SEM is arguable) do not need optics. Also, IMHO, calling the beam focus- } sing system of an electron microscope "electron optics" is an analogy with } light optics, not an identical process, so retaining the nomenclature "op- } tical microscope" to mean one with light-optic lenses is not a "mess". } } } How did we get into this mess and do we let it continue? } } } } That should stir a few minds! } } } I prefer mine shaken. } Yours, } Bill Tivol
Gee Bill,
I am not alone. It grates on me seeing that analogy turn into a definition. While I see the point of using the term LIGHT MICROSCOPE, for me electron optics are not optics. But then I am an old sh.t and one in the definite minority who still likes to play with light, and could give a whoop agout an electron. So I guess I wil just have to mumble in my single malt and let majority rule.
Shalom from Jerusalem, Azriel +++++++++++++++++++++++++++++++++++++++++++++++++++++ Major Azriel Gorski, Head Fibers and Polymers Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem, ISRAEL
azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739
What lies behind us and what lies before us are tiny matters compared to what lies within us. Ralph Waldo Emerson ++++++++++++++++++++++++++++++++++++++++++++++++++++
with Novell_GroupWise; Tue, 31 Mar 1998 08:00:29 -0600 Message-Id: {s520a29d.009-at-wpo.it.luc.edu} X-Mailer: Novell GroupWise 4.1
Hello Friends, Need your advice again! Can Spurr resin be etched like some of the Epon-type resins can for Immuno work? If so does anyone have a recipe? I have a difficult specimen, candida, that is great in Spurr, so-so in Epoxy, horrible in Lowicryl and this is getting frustrating. I don't know if Unicryl or LRwhite would work. The problem with the hydrophilic resins seems to be that the resin swells slightly as the sections are cut and the sharp distinction between the resin and the wall of the candida causes a split from the resin. The candida either fold over or fall out, leaving so few good ones that I can't proceed to immuno work and hope for any results. Thanks, Linda M. Fox lfox1-at-wpo.it.luc.edu
some of the old fashioned methods (graphical reconstruction, stereoscopy etc.) are well described in:
Gaunt, W.A. and Gaunt, P.N. (1978): Three dimensional Reconstruction in Biology. Pitman Medical Publishing Co Ltd, London (ISBN 0-272-79394-9)
Software (incl. freeware), references, links and many more you will find at:
http://biocomp.arc.nasa.gov/3dreconstruction/
Good luck
Jens
--------------------------------------------------------------- Dr. Jens Buecking Tel. +49-(0)421-218 3745 University of Bremen Fax. +49-(0)421-218 4620 Dep. of Biology Email jbueck-at-biologie.uni-bremen.de Leobener Str. - NW2 28359 Bremen ---------------------------------------------------------------
Isabel, Some years ago we needed a high surface area Al target for laser indu= ced X-ray generation. The approach we used was to evaporate aluminum through= a few Torr of N2. This produced a very granular surface. The particles were ve= ry well defined. Their nominal dimension was 100mn. It was a film but then = that is what we wanted. I suspect that grain size is a function of distance & pressure while density is a function of time. Perhaps you can tweak this approach to you need. Fair warning, this hi pressure deposition made ou= r evaporator very dirty.
Bruce Brinson Rice U.
Isabel Nogueira wrote:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } } Fellow microscopists: } } We're trying to make a deposition of 2 micron gold particles on a sili= ca } substrate. } We cannot use an evaporator because we don't want a thin film: we want = to } deposit the particles directly on the substrate, keeping them isolated = and } without forming aggregates. } At first we dispersed the powder directly on the silica, but SEM analys= is } showed they formed aggregates. } Then we've tried making a suspension on etanol and then putting a dropl= et } of the solution directly on the substrate. But still it formed aggregat= es. } } Does anyone know of a simple method to make such a deposit (without for= ming } aggregates) ? } } Thanks in advance. } } Isabel Nogueira } Instituto Superior T=E9cnico } Departamento de Engenharia de Materiais } Av. Rovisco Pais } 1096 Codex } Tel.: 351 - 1 - 8418124/0 } Fax.: 351 - 1 - 8418120 } E-mail: isabeln-at-alfa.ist.utl.pt
This may sound like a no brainer, but I am unable to find an free, PC accessible powder diffraction database (for indexing electron diffraction patterns) on the internet? So far I have found one link to SPDP-B, but I can't connect to it. Would anyone know of an alternative database?
Thanks again for your assistance.
Regards,
Michael Coviello EM Lab Manager Materials Science Dept. The University of Texas -at- Arlington Arlington, TX E-mail coviello-at-mae.uta.edu 817-272-5496
} -----------------------------------------------------------------------. } } Does anyone have experience growing tissue culture cells on gold grids } with support films? We want to do IEM of filaments after digestion of } the cytoplasmic membrane, } Sara,
I've never been able to do this very sucessfully. We've developed an approach that may do the same thing more reliably: Grow the cells on round glass coverslips, localize cytoskeletal antigens by immunogold with or without silver enhancement, mount on aluminum stubs, freeze dry, coat with aluminum, and examine with a backscatter detector in a scanning scope. Refs: Goode, D & Maugel. J.E.M. Tech. 5: 263- and Goode, D. Chap 15 In: Hayat, M.A. Immunogold-Silver Staining, CRC Press, 1995
-Dennis Dr. M. Dennis Goode Phone (301) 405-6917 Department of Zoology Fax (301) 314-9358 University of Maryland e-mail goode-at-zool.umd.edu College Park MD 20742 ************************************************************* "If the Lord Almighty had consulted me before embarking upon the creation, I should have recommended something simpler." -Alphonso X of Castile, 15th Century
} Does anyone have experience growing tissue culture cells on gold grids } with support films? } Sara Miller
Dr. Miller, You may want to consider using polystyrene (PS) films supported by gold grids to ensure attachment and growth conditions identical to those in PS flasks. You can easily dissolve a part of the PS flask in amyl acetate at ~.5%. You can float these films off from glass slides the same way as classical formavar films. I can send you our publications dealing with this and other issues of cell culture for TEM (e.g. Scan. Microscopy 1991 Sup.5: pp. S53-73 and 1996 Sup.10: pp 1-16). Marek.
Marek Malecki, M.D., Ph.D. Address: IMR, 1675 Observatory Drive, Madison, WI 53706. Phone: 6082638481 or 6084441680. Fax: 6082654076. Email: malecki-at-macc.wisc.edu http://www.wisc.edu/cesip/ http://www.bocklabs.wisc.edu/imr/integrat/transg.htm
If your antigen is not in the wall, you can digest off the cell wall and embed yeasts in anything you want. I don't know the exact recipe, but check out Laura I. Davis in Medline lit search. We did the EM for her on cells from which she had removed the CW, and the sections were beautiful. It's the thick wall that causes penetration problems.
Good luck, Sara
On Tue, 31 Mar 1998, Linda Fox wrote:
} Date: Tue, 31 Mar 1998 08:00:21 -0600 } From: Linda Fox {lfox1-at-wpo.it.luc.edu} } To: Microscopy-at-aaem.amc.anl.gov } Subject: resin etching,immuno } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Friends, } Need your advice again! Can Spurr resin be etched like some of } the Epon-type resins can for Immuno work? If so does anyone have a } recipe? I have a difficult specimen, candida, that is great in } Spurr, so-so in Epoxy, horrible in Lowicryl and this is getting } frustrating. I don't know if Unicryl or LRwhite would work. } The problem with the hydrophilic resins seems to be that the } resin swells slightly as the sections are cut and the sharp } distinction between the resin and the wall of the candida causes a } split from the resin. The candida either fold over or fall out, } leaving so few good ones that I can't proceed to immuno work and hope } for any results. } Thanks, } Linda M. Fox } lfox1-at-wpo.it.luc.edu } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Could somebody point me to the supplier/source of the ,,ready to install" attachment to the SEM stage which allows motorized moving in Z direction. This is intended to be attached to the old type of Philips XL-30 motorized stage which has manual Z movement made in the way that the stage moves NOT straight but along the circle with quite large radius - so for many applications is not critical. There is however by one of my customers the application (EBSD) where this IS very critical. I was thinking about f.e. a pallet made of piezo-crystal or smthg. The movement of 3-5 mm is enough. Not for big samples. Should have a feedtrough to outside for controlling. Appreciate any suggestions - also if somebody can make one on order in form of ,,experimental" design.
kind regards
Krzysztof Herman Labsoft, Domaniewska 9/11/45, 02-663 Warszawa tel: (48 601) 307456, tel/fx:(48 22) 483787 E-mail: kherman-at-labsoft.com.pl http://www.labsoft.com.pl/
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I tried Unicryl resin when it first came out a few years ago and did not think it had significant advantages for TEM immunocytochemistry over HM20 or LRWhite. I understand that it has been a resin "in progress" and is much improved over the original formulation.
Has anyone had recent experience with Unicryl? I am most interested in comparison with the other resins for TEM Ultrastructure (osmicated tissue) and/or immunocytochemical localization. I want to use it primarily for plant tissue and cyanobacteria so do need a resin with low viscosity for adequate infiltration of tough cell walls. I will want to be able to polymerize with either UV light or heat depending on whether I am using freeze-substituted material or chemical fixed material.
Thanks in advance, Debby Sherman, manager Microscopy Center in Agriculture Purdue University
id sma010745; Tue Mar 31 23:29:31 1998 Received: from c1pa008.lkasbg.gv.at (c1pa008.lkasbg.gv.at [10.1.42.8]) by hermes.lkasbg.gv.at (8.8.5/8.8.5) with SMTP id AAA14191; Wed, 1 Apr 1998 00:31:57 +0200 Received: by c1pa008.lkasbg.gv.at with Microsoft Mail id {01BD5CFD.4920A780-at-c1pa008.lkasbg.gv.at} ; Tue, 31 Mar 1998 23:32:35 +-200 Message-ID: {01BD5CFD.4920A780-at-c1pa008.lkasbg.gv.at}
Hi Linda,
Most recipes for deplastification of resin sections have been published = for EPON, but (thanks God for my being busy in former times in reading = original articles and listing keywords for the contents) maybe some = keywords would work for you and your request:
MOTHES-WAGNER U, WAGNER G, REITZE HK, SEITZ KA: A Standardized Technique for the *in toto* epoxy resin embedding and = precipitate-free Staining of Small Specimens covered by Strong = Protective outer Surfaces. J. Microscopy 134, 307-313, 1984: =20 Embedding: Modified Spurr's for cuticularized specimens (small), i.e. = 26g NSA, 10g ERL 4206, 8g DER 736, 0.2g S-I) Removal: by Sodium-methylate solution (sodium-methoxide)
PEDRAZA MA, MASON D, DOSLU FA, MARSH RA, BOBLETT JP: Immunoperoxidase Methods with Plastic Embedded Materials Laboratory Medicine 15, 113-115, 1984 Embedding: Araldite, Spurr's, (M-)methacrylate Removal: Sodium-ethoxide (sodium-ethylate) according to=20 MAYOR HD, HAMPTON JC, ROSARIO B: A simple Method for Removing the Resin from Epoxy-Embedded Tissue J. biophys. & biochem. Cytology(Baltimore) 9, 909-910, 1961 (Cave: uses Toluol/benzole/benzene/metallic Na- mixtures)
RUSSELL SD, DAGHLIAN ChP: Scanning Electron Microscopic Observations on Deembedded Biological = Tissue Sections: Comparison of Different Fixatives and Embedding = Materials. J. Electron Microscopy Technique 2, 489-495, 1985 (great micrographs!)
Embedding: Paraffin, SPURR's resin; Removal: Using the Method described by=20 HOGAN DC and SMITH GH: Unconventional Application of Standard Light and = Electron Immunocytochemical Analysis to Aldehyde-fixed, = araldite-embedded Tissues: J. Histochem Cytochem 30: 1301-1305, 1982 means ("alla breve"): } } dry specimen: placed in freshly mixed 0.5% KOH = in (absolute? yes! I would take this one!) methanol mixed in a 1:2 = ratio with a solution of 1:1 benzene and acetone for 5-10 min ( back to = stone age!). KOH was neutralized by a 1-min immersion in 1% acetic acid in = *absolute*(here it is!) methanol, and rinsed in absolute methanol. This = formulation is also available (TEXT 1985!!) in a premixed Kit from = POLYSCIENCES, Warrington PA at approx. US$ 35.-. Although Hogan and = Smith(1982) recommend longer times than those employed in the present = study, the SEM results suggest that extraction was completed within the = time periods listed above. Hydrogen peroxide oxidation is used in the = HOGAN and SMITH(1982) technique to remove Osmium salts (personal note = added: this oxidation erroneously mostly referred today as "etching" = sections for use in ICC/IHC(IMM-EM)), but was omitted in the present = (1985) procedure. { {
} From a Poster Text at the Histochemical Society's Symposium at = GARGELLEN/AUSTRIA 1986 (unfortunately no exact citations present) TAATJES....., ROTH J. (Basel), from a poster in workshop: LECTINS: = Methods and Applications, Silver amplification, etc:=20 my own notices: Embedding: EPON, Removal: for LM-Incubation for LECTIN-Gold-Complex: MAXWELL's Fluid: i.e. 2 g KOH, 5 ml Propylene-oxide, 10 ml Methanol abs = mixture: 2-3 min (on semithin sections, 1-2 ?m), followed by MetOH/PBS = 5 min, then PBS 2x5 min. (maybe this variant would work on Spurr's = too??)
IWADARE T, HARADA E, YOSHINO S, ARAI T.: A solution for removal of Resin from Epoxy Sections Stain Technology (now: BIOTECHNIC and HISTOCHEMISTRY) 65/#4, 205-209, = 1990 Embedding: EPON Removal by A MIXTURE of CROWN-ETHER (18-CROWN-6 (SIGMA No C 5515) DMSO, = K-methoxid in MetOH using LOW ALCALI CONCENTRATIONS.....Incubation for = approx. 5 min up to 30 min (depending on section thickness): would = perhaps work also on SPURR's
Van PELT-VERKUIL E: Detection of Labile and Low Concentrations of Antigens in Semithin = Resin-Sections (Technovit 8100). UK-NEQUAS-ICC(Official Newsletter of the UK National External Quality = Assessment Scheme for Immunocytochemistry), #5, Summer 1995, p. 10-16: my own notice(s): stated in the text there: } } LR WHITE & LOWICRYL PLASTIC in Semithin = Sections CANNOT be removed.
Dear Linda: unfortunately I did such experiments a long while ago. I had = some success then but would have to establish the whole on a proper, = "today's" science. Hope my info's could be of help to you and your "nasty Job on resin = removal" (especially concerning Spurr's). I think you would have to test = a lot of sections before, if those with incorporated "candida" would be = very precious and "solitaires".
Note added "in proof": be sure, if working with the highly alkaline = solutions (KOH, Methylates, Ethylates) you rinse the sections after = Resin-removal carefully and extensively first in the solvent "sister" = (i.e. MetOH abs, EtOH abs, then A. bidest, then buffer) to get rid of = the long sticking alkaline solution as well as the depolymerization = product remnants: those would interfere, at least in certain areas of = the sections, with successful IHC-staining with your antibodies. Best wishes
Wolfgang
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
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Hello Friends, Need your advice again! Can Spurr resin be etched like some of the Epon-type resins can for Immuno work? If so does anyone have a recipe? I have a difficult specimen, candida, that is great in Spurr, so-so in Epoxy, horrible in Lowicryl and this is getting frustrating. I don't know if Unicryl or LRwhite would work. The problem with the hydrophilic resins seems to be that the resin swells slightly as the sections are cut and the sharp distinction between the resin and the wall of the candida causes a split from the resin. The candida either fold over or fall out, leaving so few good ones that I can't proceed to immuno work and hope for any results. =20 Thanks, =20 Linda M. Fox=20 lfox1-at-wpo.it.luc.edu
--- Received from HQ02MEMO.TC6NBLD DAVIS, BONNIE 03-31-98 16:55
I am currently evaluating image capture and analysis systems for metallography at production locations. I have information regarding the PAX-IT System sold by Midwest Information Systems. It is a hardware and software solution, and I am hesitant about purchasing what I consider to be somewhat of a 'black box'. Midwest Information Systems appears reluctant to specify what hardware they use, other than to say that the hardware I currently use (Matrox Meteor frame grabbers) is not supported. However, a single package system may have benefits for multiple users at geographically distant sites. Does anyone have any opinions or experience with the PAX-IT system from Midwest Information Systems?
Thanks Bonnie Davis Staff Engineer Materials Analysis Kennametal, Inc. Route 981S Latrobe, PA 15601
---- 03-31-98 16:55 ---- Sent to --------------------------- -} IBMMAIL.INTERNET Advantis IBM Mail Exchange
} Folks: } We are trying to automatically measure slopes on the sides of features protruding an otherwise smooth surface imaged by 3-D profilometry. The data set is comprised of x,y position vs. z height. The x and y data } are evenly spaced but not necessarily by the same amount. If anyone has or } knows of a canned routine/plug-in etc. that would help us out, please send me } a note. We can handle software for almost any platform } (Mac/Windows/Unix/VAX/etc.) } } Bill Heeschen } The Dow Chemical Company } 517-636-4005 } waheeschen-at-dow.com
I am in need of a type 545 Polaroid camera back that has a frosted glass where the film holder usually is. My SEM pix look OK on the screen but if I make a print from the neg. it looks slightly out of focus. I've been told that I need to focus the camera to match the screen & this fancy camera back with the glass helps you do that. I know they exist because I saw one on an ESEM. Does anyone know where I can buy one? I called Polaroid and they were confused. Can anyone help me with this problem?
My future looks fuzzy & so do my prints,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Alex- we are becoming fairly proficient in 3-d recon here in the Kinnamon lab. what types of computers do you have access to? and what type of TEM data? tomographic or serial sections? the rest is ....easy (after a lot of practice). there are many pieces of software avail at low cost. -Mike
On Tue, 31 Mar 1998, Alex Black wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear listees, } I want to do some 3 dimensional reconstruction of a particular } type of fibroblast in normal and pathological states, but I really am } at a loss as to the methodology. I do not have access to a program to } do this, so I need some sort of 'old-fashioned' but reliable method. } Any help will be gratefully receieved (as will any cheques, } real-estate, cars, post-doc offers etc. etc.). } Thanks in advance, } Yours 2 dimensionally, } Alex } } } } ...................................................................... } ....................................... } Alex Black } Department of Anatomy } National University of Ireland } Galway } } alexander.black-at-ucg.ie }
Can someone tell me a bit more about Unicryl? I'm not a biologist, I work in a Geology department, would love a non- viscous resin which would soak well into crumbly, porous rocks and wouldn't polymerise until told to. Who makes/supplies it?
Is it an acrylate resin? How runny is it? How does one initiate the polymerisation? How long is the pot life at room temperature? etc etc
thanks
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Micro Analytical Laboratories, Inc. is a commercial laboratory, specializing in environmental analyses of air, water, soil, and building materials. One position is open for full-time employment.
The applicant should possess strong communication and analytical skills. Optical mineralogy or electron microscopy coursework is preferred.
Please FAX or mail a resume to:
Frank Raviola Micro Analytical Laboratories, Inc. 5900 Hollis St., Suite M Emeryville, CA 94608
At 02:45 PM 3/31/98 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Paula,
When I had to do this with a Hitachi S570 camera, I took the ground-glass from a Polaroid MP-3 copy camera. You know the kind that takes 4x5 film and has a copystand with lights? If you can find an on-campus facility that does document copying, photographic copying, etc., such an instructional learning resources center, they may have one. They may also have something else that might work. Look for facilities like "research photography", "scientific photography", "photographic services", "photo production technology", and other such names. Even if they're not using such technology anymore, in these days of digital imaging, you can bet that somebody has a 4x5 ground glass attachment in a drawer or cupboard somewhere. If there is a photography department there, they will certainly have one around.
Failing everything else, there is an old trick using frosted adhesive tape. Stretch it across the camera opening for the film and it will serve as a ground glass substitute. You MUST, however, make sure that the tape is in the exact same plane as the film in your holder would be, and that part will rely on you and a good ruler and imagination. The other thing, of course, is to position the tape so that the photo CRT projects onto it during the photo capture time. Probably it will require a few strips of tape to give you enough time to adjust the camera during the exposure time. Use a loupe to focus the projected beam onto the tape, being careful not to bend the tape in the process. An acrobatic feat, to say the least.
Good luck.
Randy
Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
Have you ever seen the paper studying about the diffusion rate of dyes in food systems(microscopically observed)? How can I search for those kinds of materials? Thanks in advance Masubon Thongngam
Maybe you are referring to the Mamiya focussing screen holder (# Y5.500?) which is used to set up the Mamiya Press 6 x 7 120 roll film camera?
Our JEOL 35C SEM and JEOL 200CX TEMSCAN were set up with this for the roll film cameras which are supposed to be interchangeable with the Polaroid 545 film holder. I think they worked out ok, but we could never afford to run too much Polaroid!
Further to Randy's suggestions: use a long shutter time if you can- it helps! Also, I believe I have done the job looking at the waveform rather than a normal image, because you're looking at something you know.
Hi, can anyone tell me about experience with anti-gfp antibodies on glutaraldehyde fixed cells or tissue? I am looking for an antibody which is suitable for electron microscopical gold labelling. Someone out there who has already tried it with or without success? thanks reinhard . . . . . . . . . . . . . . . . . . . Dr. Reinhard Windoffer Fon: (00)49 (0)6131/39 3720 Universitaet Mainz Fax: (00)49 (0)6131/39 4615 Anatomisches Institut e-mail: windoff-at-mail.uni-mainz.de Becherweg 13 D-55099 Mainz Germany . . . . . . . . . . . . . . . . . . .
we have been informed that You work on the structural investigation of proteins by high resolution electron microscopy. According to recently found unpublished notes traceable to Ernst Ruska the resolution limitation of a transmission electron microscope due to spherical aberration can be overcome by placing the microscope into a highly inhomogenous magnetic field such as at the poles of the larger planets of our solar system. Additionally one can expect extremely stable cryo-conditions around 4 Kelvin in the outer regions of our solar system. The National Aeronautics and Space Agency of the United States of America has therefore launched a project to install a Philips CM200FEG inside an unmanned satellite to be sent to Jupiter where it will be held in orbit for the duration of the experiment. All manipulations such as alignment and specimen insertion will be carried out by a remotely controlled robotic arm.
We are currently seeking suitable specimens for these undertaking and would appretiate contributions from Your department in exchange for a co-authorship in all resulting publications.
I wish to receive notice about the book "Manual of microscopic Analysis of Feedstuffs. 3rd Ed. The Amer. Assoc. of Feed Microscopists, 1922, p. 73-93" or the articles in the same book, autors BATES L.e coll. Feed ingredient descriptions of animal origin.
Hello everyone, I am trying to make cross-section TEM samples of thin films (ZnO) on sapphire. So far I have tried to make the samples by the traditional method ie., glue two pieces face-to-face, polish to 15 micron, and then ion-mill. I have had some problems with this method. Some of the people I have contacted about this told me to try using the wedge polishing method. Has anyone out there used this technique for making cross-section samples of films on sapphire? I was informed by another person that this might not work as sapphire is very brittle when it is thin, and we might not get a very good wedge. I would greatly appreciate any hints that you can give.
Sincerely
Chandrasekhar (chandu) Gorla Rutgers University Dept. of Ceramic and Materials Science Piscataway, NJ 08855 TEl: (732)445-3663
____________________________________________________________________ Get free e-mail and a permanent address at http://www.amexmail.com
Unicryl is made by BBInternational and distributed in the USA by Vector Laboratories, Inc. (1-800-227-6666). They are a CA based company and are not open yet today. I did call there yesterday and asked for a method booklet on Unicryl and the individual who I talked to did not indicate that the resin was unavailable. In fact, I just recieved a promotional brochure on the resin yesterday which prompted me to look into it further.
For those that are not familiar with Unicryl, it is a resin that is supposed to be more all purpose. The company claims that it is good for light and electrom microscopy, immunolabeling, in situ hybridization and histochemistry. It is a one component resin,of low viscosity and can be polymerized with UV light at 4o C or at 55oC.
This all sounds good but I did want to benefit from others experience as to how it compares with other resins such as Epon generic, Spurr's, LR White, and the various lowicryls like HM20 (my favorite for low temp.ICC).
Debby Sherman, manager Microscopy Center in Agriculture Purdue University
--------------------------------------
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Dear Walter I feel there must be an easier way! However, I have despatched for your personal attention a live sample of the new mutant AIF strain of Ebola virus, which has a novel and enigmatic structure we are anxious to elucidate. I assume NASA has the appropriate containment resources to deal with the consignment on arrival. Although I don't expect to hear from you again immediately, I would appreciate it if you could keep me informed of progress with the experiment. Chris
------ Forwarded Message Follows -------
Hello Friends, Need your advice again! Can Spurr resin be etched like some of the Epon-type resins can for Immuno work? If so does anyone have a recipe? I have a difficult specimen, candida, that is great in Spurr, so-so in Epoxy, horrible in Lowicryl and this is getting frustrating. I don't know if Unicryl or LRwhite would work. The problem with the hydrophilic resins seems to be that the resin swells slightly as the sections are cut and the sharp distinction between the resin and the wall of the candida causes a split from the resin. The candida either fold over or fall out, leaving so few good ones that I can't proceed to immuno work and hope for any results. Thanks, Linda M. Fox lfox1-at-wpo.it.luc.edu
Hi Linda,
We used Unicryl resin for many years on several yeasts with good results. We do not remove the cellwall of the cells but "etch" it a little by an incubation with 0.1% meta periodate for 5 minutes prior to the dehydration. See the paper of Evert van Tuinen Journal of Histochem. & Cytochem. vol. 35/3, page 327-333 (1987).
You should also I have filed a formal complaint to the originator's postmaster of this message since it was signed by a bogus account from "rumble.csb.ki.se" and was sent in a way to attempt to both hide detection and also illegally represented an agency of the US Government.
It is obvious this was a joke, but it broke all the Listserver Rules, which everyone agrees to when they sign on.
Jokes I can put up with, but not changing addresses, attempting to hide and impersonating a government agency!
Does anyone know where one can find 100kV DC Cables? Multicore cables that will hold off the 100kV?
Your assistance is appreciated.
-Karl
-- "It wouldn't be research... if I knew what I was doing." +++++++++++++++++++++++++++++++++++++ ++ Karl R. Umstadter, Ph.D. ++ Los Alamos National Laboratory ++ P-23, PFX-I Research ++ MS H803, Los Alamos, NM 87545 ++ Office: 505.665.1509 ++ Fax: 505.665.4121 +++++++++++++++++++++++++++++++++++++
We have been using a LEO 435 VPi SEM with an Oxford Inst. ISIS 300 integrated system since December of 1996. Since day one, we have had a series of problems with this integrated system, apparently caused by the common operating systems (first Win 3.1, then WIN95). It appears that the two systems do not like being integrated under one computer, as the sharing of resources seems to cause lurking problems that randomly appear, and cause system failures. I would be interested if any other users of the ISIS 300 system have had any other similar problems. Oxford claims that we are alone with our troubles, but I find this very hard to believe.
The first incarnation of our system used WIN 3.1 as the operating environment. This seemed optimum for the LEO as we encountered no system problems here. The ISIS though, would constantly run out of system resources and crash, usually without warning and lock up the system. It turned out that the ISIS is a resource hog (perhaps due to being written in Visual Basic), and the upgrade to ver 3.2 did not help at all. We then upgraded the system to Win 95, and the resource problem with the ISIS appears cured, but we have uncorked a beast in the LEO. The LEO is being upgraded to a full win95 system soon, but I am seeing more and more bugs with the ISIS.
Am I alone in believing the ISIS to be a poorly conceived and executed system? We have had endless conflicts with the operating system, to the point of having to reinstall win95 several times after apparent simple crashes, which seem to render several ISIS modules unavailable after that. The user interface appears haphazard from module to module, with no user definable defaults to customize operation to our preferences. The Labbook system forces users to collect data according to the x-ray system parameters. We like to structure our data by customer and job parameters, and keep all x-ray and microscope data under one directory. This is impossible under the ISIS Labbook system.
I am greatly disappointed with the integration of these two products. The LEO has been a fine system, with the operating environment keyed to usability, and endlessly customizable by the user. The ISIS is user unfriendly, and rigid in structure. Unless the user follows the Labbook protocols exactly, the system is unavailable to them.
Has anyone heard of a replacement operating environment for the Oxford system? Our complaints to Oxford have been dismissed, and LEO is unable to effect changes to the abysmal Oxford operating system. I wonder if I am alone in my frustration with this unfriendly and quixotic x-ray system.
Mike Warfield Hughes Space & Communications M&P Laboratory
You didn't say if your were dimpling the sample before ion milling and you didn't say what your ion milling conditions were -these are important. 15 um is a lot of material to ion mill through and you can minimize or eliminate differential sputtering effects by using low angles. You should precision dimple the sample down to less than 5 um and you can probably get close to 1 um. Then ion mill. If you have an older ion mill, you are going to need a sector control stage. If you have a newer ion mill, use low angles but remember that you have to clear the rim of the dimple. This means that if you have a 100 um thick sample and dimple on one side, you can't get below about 4 degrees. Double-sided dimpling and thinner sample allows you to go lower.
You can do the Tripod technique but it takes an investment in time and some equipment to learn it. It is best to learn from someone who knows. South Bay Technology has a periodic short course taught by Ron Anderson. You should contact them to see what is involved.
Sapphire also works with the Small Angle Cleavage Technique. I've done ZnO on Si and glass coverslip substrates this way and it works well. You should be able to do it with sapphire easily enough. This technique has a lot of advantages, but one of the best is that it is very cheap. South Bay Technology has a kit for getting started on this technique called the MicroCleaving Kit. John McCaffrey and I have a detailed pictorial tutorial on how to do it in the "Specimen Preparation for TEM of Materials IV" book Vol 480 from MRS that should be able to get you started on it. I always try this technique first; if it works, it's great.
Good luck.
Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Hello everyone, I am trying to make cross-section TEM samples of thin films (ZnO) on sapphire. So far I have tried to make the samples by the traditional method ie., glue two pieces face-to-face, polish to 15 micron, and then ion-mill. I have had some problems with this method. Some of the people I have contacted about this told me to try using the wedge polishing method. Has anyone out there used this technique for making cross-section samples of films on sapphire? I was informed by another person that this might not work as sapphire is very brittle when it is thin, and we might not get a very good wedge. I would greatly appreciate any hints that you can give.
Sincerely
Chandrasekhar (chandu) Gorla Rutgers University Dept. of Ceramic and Materials Science Piscataway, NJ 08855 TEl: (732)445-3663
____________________________________________________________________ Get free e-mail and a permanent address at http://www.amexmail.com
We look after a number of Topcon / ISI systems and find that we have a repeat problems of the Power Transistors for the Bias, filament and EHT control blowing. These are the four transistors that sit in the small box on top of the EHT tank on the Sx SS and ABT range of E.M.'s We have tried a number of transistors and ways of mounting these, but they continue to give problems. Does anybody else have the same problem, but more importantly has any one solved this problem ?
Thanks Luc Harmsen Anaspec, South Africa Technical support for E.M. operators, world wide. anaspec-at-icon.co.za TEL: ++ 27 (0) 11 476 3455 FAX: ++ 27 (0) 11 476 7290
The presence of microbes in the deep core is significant to considerations of life elsewhere. See the story for more information.
Scott
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
At 11:59 AM 4/1/98 +0100, you wrote: } Received: from rumble (rumble.csb.ki.se) by ondine (4.1/SMI-4.1) -=-snip-=- } Dear fellow researcher, } -=-snip-=-
Those wacky jokesters at the Department of Bioscience of the Novum Karolinska Institutet in Sweden. Always good for a few laughs. ;} )
} Can someone tell me a bit more about Unicryl? } I'm not a biologist, I work in a Geology department, would love a non- } viscous resin which would soak well into crumbly, porous rocks and } wouldn't polymerise until told to.
Ritchie -
I suggest that you try LR White, Hard grade. It works well for things like zeolite catalysts, for example. Check these refs for details:
Csencsits, R., Schooley, C.,& Gronsky, R. (1985) An improved method for thin sectioning of particulate catalysts. J.E.M. Technique 2:643-644.
Ulan, J.G., Schooley, C., & Gronsky, R. (1990) Microtomy of large particle zeolites for TEM. Mat. Res. Soc. Symp. Proc. 199:153-156
Ulan, J.G., Schooley, C., & Gronsky, R. (1990) Modified embedment procedure for microtomy of large particle zeolites. J.E.M. Technique 16:254-255
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
I would suggest that we could examine the Good Times virus or we may still have some Pilt Down Man bones in the basement.
Thanks for the opportunity to collaborate but I would prefer the air miles if NASA is in that scheme.
Malcolm Haswell University of Sunderland UK ----------
Dear fellow researcher,
we have been informed that You work on the structural investigation of proteins by high resolution electron microscopy. According to recently found unpublished notes traceable to Ernst Ruska the resolution limitation of a transmission electron microscope due to spherical aberration can be overcome by placing the microscope into a highly inhomogenous magnetic field such as at the poles of the larger planets of our solar system. Additionally one can expect extremely stable cryo-conditions around 4 Kelvin in the outer regions of our solar system. The National Aeronautics and Space Agency of the United States of America has therefore launched a project to install a Philips CM200FEG inside an unmanned satellite to be sent to Jupiter where it will be held in orbit for the duration of the experiment. All manipulations such as alignment and specimen insertion will be carried out by a remotely controlled robotic arm.
We are currently seeking suitable specimens for these undertaking and would appretiate contributions from Your department in exchange for a co-authorship in all resulting publications.
by pohl.acpub.duke.edu (8.8.5/Duke-4.5.3) with ESMTP id LAA23957; Wed, 1 Apr 1998 11:10:41 -0500 (EST) Received: (from saram-at-localhost) by soc11.acpub.duke.edu (8.8.5/Duke-4.4) id LAA00765; Wed, 1 Apr 1998 11:10:40 -0500 (EST)
Refs for virus pix:
Palmer EL, Martin ML. Electron Microscopy in Viral Diagnosis. CRC Press, Boca Raton. 1988. (Ebola in here)
Doane FW, Anderson N. Electron Microscopy in Diagnostic Virology. Cambridge Press, New York. 1987. (Marburg here)
Miller SE. Detection and identification of viruses by electron microscopy J Electron Microsc Tech (now J Microscopy Research and Technique) 1986;4:265-301.
Miller SE. Diagnosis of viral infections by electron microscopy. In Lennette EH, Lennette DA, Lennette ET (eds). Diagnostic PRocedures for Viral, Rickettsial, and Chlamydial Infections. American Public Health Assoc, Washington, DC. pp 37-78. (Ebola here)
Murphy FA;, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, Martelli GP, Mayo MA, Summers MD (eds). Virus taxonomy, 6th Report of the International Committee on Taxonomy of Viruses. Springer-Verlag, New York. (Marburg and Ebola here0
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We use a simple ground glass screen with a magnifying lens built into a plastic housing which fits on the back of the camera in place of the Polaroid 545 back. It was cheap, and came from our local photographic supply house. It works fine.
Tony Garratt-Reed
At 02:45 PM 3/31/98 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
We use LRWhite alot. The stains rinse out very quickly. Use just 10 dips in 2 beakers of distilled water. then dry.
Bob Underwood Derm Imaging center
On Fri, 27 Mar 1998, Barbara Plowman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have been staining some LRWhite(medium) sections and I am getting very low } contrast with 30min. 3%UA and 10 min. Lead Citrate. I have tried reducing the } wash times and the KV to 60KV without any luck. Any ideas and/or advice? } } Barbara Plowman } University of the Pacific } School of Dentistry } 2155 Webster } San Francisco, CA 94598 } email:Bplowman-at-uop.edu }
In our diagnostic EM lab we currently use Paragon stain to stain our epoxy semi-thin sections, but it would be very nice if we had some kind of rapid H&E like stain that could give that kind of polychrome differentiation between the nucleus and the cytoplasm.
The Paragon stain is supposed to be a polychrome stain, but what we in effect see is more of a monochrome direct stain.
Does anyone know of a quick yet superior stain that might give us better results here?
What I would like to know is how Philips scored this cherry deal= to get their scope in orbit, and, in what way they reduced costs= in order to get below NASA's ceiling for non-competitive action?= Considering the application I suppose the vacuum pump system could= be deleted: Who needs a rotary pump when for backing vacuum you= can "vent to space"...
The question is would Balzers, Edwards and other members of the High= Vacuum Equipment Consortium initiate a Congressional Oversight Committee= Investigation into the matter? If so, our samples destined to be= launched with the probe could be lost for a decade or so in this= legislative morass!
Even with the reduction in maintenance-requiring systems did NASA= obtain a service contract through Philips or, in an effort to contain= costs, get one with another third party service provider? I pity= the poor service technician whose service region this unit falls,= or rather orbits, in. Then again, just think of the mileage reimbursement.= =20
The question lingering, however, will be if FedEx will be able to= deliver replacement parts still by P1 Overnight Express...
************************************************************ It's true- the inmates ARE running the asylum... ************************************************************ Laura Rhoads Electron Microscopy Facility Director Department of Biology Western Kentucky University 1 Big Red Way Bowling Green, KY 42101-3576
Folks, do any of you know of a supplier for ITO glass for making heated culture chambers Thanks Simon
----------------------------------------------------------------------- Simon C. Watkins Ph.D. Associate Professor Director, Center for Biologic Imaging University of Pittsburgh Pittburgh PA 15261 tel:412-648-3051 fax:412-648-2004 URL: http://sbic6.sbic.pitt.edu
Does any one have any info on a CITZAF type package that includes a thin film correction and is NOT CITZAF3 by John Armstrong?? We are interest in anything, Commercial or Freeware.
I am not currently part of the list, so please mail to me directly.
I am currently in the process of restoring a Zeiss Axiomat LM. We do not have the screw rings/adapters required for attaching a camera, and are still waffling over film vs. digital cameras. If I go with with film, do I absolutely need a low momentum shutter? Can anyone give me advice on where to look for the adapters? I know that it will be difficult to find parts for a twenty-something year old scope, but it seems to be in good working order. Thanks for any help
Charlie Ginsburg NCC Research Dept. Lombard IL cgins-at-yahoo.com
_________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
Hi, I just received a job announcement from the Mayo Clinic in Rochester, MN. by snail mail. They are looking for an Electron Microcopy Technologist to work at their Core Facility . Please contact Kaine Kerkhoff {kerkhoff.kaine-at-mayo.edu} if you are interested in this position. thought some of you might like to know, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------= =2E } =
} Fellow microscopists: } =
} We're trying to make a deposition of 2 micron gold particles on a sili= ca } substrate. } We cannot use an evaporator because we don't want a thin film: we want = to } deposit the particles directly on the substrate, keeping them isolated = and } without forming aggregates. } At first we dispersed the powder directly on the silica, but SEM analys= is } showed they formed aggregates. } Then we've tried making a suspension on etanol and then putting a dropl= et } of the solution directly on the substrate. But still it formed aggregat= es. } =
} Does anyone know of a simple method to make such a deposit (without for= ming } aggregates) ? } =
} Thanks in advance. } =
} Isabel Nogueira } Instituto Superior T=E9cnico } Departamento de Engenharia de Materiais } Av. Rovisco Pais } 1096 Codex } Tel.: 351 - 1 - 8418124/0 } Fax.: 351 - 1 - 8418120 } E-mail: isabeln-at-alfa.ist.utl.pt
There are reliable methods to make coloidal gold in suspension. The gold particles are very regular in size and show little aggregation. Also aggregates can be removed by centrifugation. Your problem would seem to be to deposit these particles onto the silica substrate without aggregation. I guess it should not be impossible by adjusting concentration and charge of the particles in suspension. Also, they probably adsorb to the film if you put the suspension briefly in contact with the substrate. If everything fails, may be they can be centrifuged into the silica. It is possible that the coloidal gold is more stable in solution than the gold you are using, but I do not know if it is possible to make it as large as you want. Usually the particles have up to 15nm. May be they can be grown after they are deposited in the silica.
Good luck
A.P. Alves de Matos Electron Microscopy Unit Curry Cabral Hospital Lisbon
I make a focusing screen for a homemade "roberts lens focuser" for a nikonos underwater camera, by grinding a 1.5" x 3" microscope slide using some grinding compound. Worked fine for that purpose, but you'd have to determine the plane of focus and make sure the glass you use will span the width. It may not have to cover the entire surface.
Alan Davis adavis-at-netpci.com
On Wed, Apr 01, 1998 at 12:22:11PM -0500, Tony Garratt-Reed wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Paula - } } We use a simple ground glass screen with a magnifying lens built into a } plastic housing which fits on the back of the camera in place of the } Polaroid 545 back. It was cheap, and came from our local photographic } supply house. It works fine. } } Tony Garratt-Reed } } At 02:45 PM 3/31/98 -0800, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Listers, } } } } I am in need of a type 545 Polaroid camera back that has a frosted } } glass where the film holder usually is. My SEM pix look OK on the screen } } but if I make a print from the neg. it looks slightly out of focus. } } I've been told that I need to focus the camera to match the screen } } & this fancy camera back with the glass helps you do that. I know they } } exist because I saw one on an ESEM. Does anyone know where I can buy one? } } I called Polaroid and they were confused. } } Can anyone help me with this problem? } } } } } } My future looks fuzzy & so do my prints, } } } } Paula :-) } } } } Paula Sicurello } } UC Berkeley } } Electron Microscope Lab } } psic-at-uclink4.berkeley.edu } } } } } } } } } Anthony J. Garratt-Reed } MIT Room 13-1027 } 77 Massachusetts Avenue } Cambridge, MA 02139-4307 } United States of America } } Ph: 617-253-4622 } Fax: 617-258-6478 } }
-- Alan E. Davis Marianas High School (Science Department) AAA196, Box 10001 adavis-at-netpci.com http://www.saipan.netpci.com/~adavis Saipan, MP 96950 15.16oN 145.7oE GMT+10 Northern Mariana Islands
By GFP, I presume you mean glial fibrillary acidic protein. If you don't, ignore the rest of this! If you do, then you're in luck. GFAP is one of the toughest antigens there is, surviving in OsO4 fixed tissue, embedded in resin and left in a cupboard for 15 years. I've done a lot of labelling for GFAP. The best antibody I've tried is Dako's polyclonal rabbit anti-cow. Others I've tried include Amersham monoclonal mouse anti-human and Sigma polyclonal rabbit; they weren't as effective. You can label glut fixed whole tissue - assuming, of course, that you either use vibratome sections or permeabilise the tissue. You can label any old resin sections - if you can see the filaments, then there should be enough GFAP to label. Pretreat with NaIO4 first, then immunolabel as usual. If you're interested I can send you the method(s) I use. I've labelled with 10nm gold conjugate and also 1nm with Ag enhancment. Both methods work well.
Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
It is important that the ground part of the glass is precisely in the focal plane. The ground glass should be facing the lens and sit on the film guides. A ground glass screen is easily made by grinding the face of a cut (perhaps larger than standard) microscope slide. Say 800 grid grinding compound (as used for grinding geol. section) does the job very well.
First the magnifier should be focused onto the ground screen.
Then press photo (no beam required) and use a line scan at low intensity as an "image" on the screen.
Adjust the distance between CRT and camera to bring the image into focus. Alternatively, the lens on the camera can be focused.
Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
____________________________________________ } We use a simple ground glass screen with a magnifying lens built into a } plastic housing which fits on the back of the camera in place of the } Polaroid 545 back. It was cheap, and came from our local photographic } supply house. It works fine. } } Tony Garratt-Reed } } At 02:45 PM 3/31/98 -0800, you wrote: ----------------------------------------------------------------------. } } } } Listers, } } } } I am in need of a type 545 Polaroid camera back that has a frosted } } glass where the film holder usually is. My SEM pix look OK on the screen } } but if I make a print from the neg. it looks slightly out of focus. } } I've been told that I need to focus the camera to match the screen } } & this fancy camera back with the glass helps you do that. I know they } } exist because I saw one on an ESEM. Does anyone know where I can buy one? } } I called Polaroid and they were confused. } } Can anyone help me with this problem? } } } } } } My future looks fuzzy & so do my prints, } } } } Paula :-) } } } } Paula Sicurello } } UC Berkeley } } Electron Microscope Lab } } psic-at-uclink4.berkeley.edu
INTERNATIONAL COURSES OF LIGHT MICROSCOPY, PHOTOMICROGRAPHY AND LASER SCANNING CONFOCAL MICROSCOPY GARGNANO (Lake of Garda) October 1998
The Course is a post-graduated theoretical/practical course, with propedeutical lectures and practical stages on microscopy, photomicrography and confocal microscopy. The course will take place in Gargnano (Lake of Garda) in October 1998.
All information and registration details (participation fee, date, special accomodation) at the following Web address.
http://imiucca.csi.unimi.it/endomi/micro.html
Thank you Paolo Castano
_____________________________________________________ Prof. Paolo Castano UNIVERSITY OF MILAN INSTITUTE OF HUMAN ANATOMY - CHAIR OF HUMAN ANATOMY FOR PHARMACY Via Mangiagalli, 31 - 20133 Milan (Italy)
Hello Friends, Need your advice again! Can Spurr resin be etched like some of the Epon-type resins can for Immuno work? If so does anyone have a recipe? I have a difficult specimen, candida, that is great in Spurr, so-so in Epoxy, horrible in Lowicryl and this is getting frustrating. I don't know if Unicryl or LRwhite would work. The problem with the hydrophilic resins seems to be that the resin swells slightly as the sections are cut and the sharp distinction between the resin and the wall of the candida causes a split from the resin. The candida either fold over or fall out, leaving so few good ones that I can't proceed to immuno work and hope for any results. Thanks, Linda M. Fox lfox1-at-wpo.it.luc.edu
Hi Linda,
We used Unicryl resin for many years on several yeasts with good results. We do not remove the cellwall of the cells but "open" it a little by an incubation with 0.1% sodium meta periodate (w/v in water) for 5 minutes prior to the dehydration. See the paper of Evert van Tuinen Journal of Histochem. & Cytochem. vol. 35/3, page 327-333 (1987).
Nestor J. Zaluzec wrote: } } Enough said ! } } You should also I have filed a formal complaint to the originator's postmaster } of this message since it was signed by a bogus account from } "rumble.csb.ki.se" and was sent in a way to attempt to both hide detection } and also illegally represented an agency of the US Government. } } It is obvious this was a joke, but it broke all the Listserver Rules, which } everyone } agrees to when they sign on. } } Jokes I can put up with, but not changing addresses, attempting to hide and } impersonating a government agency! } } Nestor
Dear Nestor and list members and NASA,
I'm sorry about any bad feelings I've caused and I'm turning myself in.
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
The problem you describe is something we saw with a S440i system about 3 years ago. As is usual, the SEM manufacturer and the ED manufacturer will both say the intergration works before you place the order and then both deny it is their system crashing after you have paid. ( please don't sue me for this !!!!??? ) Sorry to say it but you must simply de-integrate the two. Isis was designed around the HP type PC. So let it run on that. The Leo will run happily on the PC. We must remember that P.C. stands for Personal Computer. So documents and games work ......reasonably well !!???....... But if you load a P.C. with real work, it gets a bit of a speed wobble and crashes fairly often. The Leo, on it's own, should have 64Mbytes of memory fitted to make it work comfortably and ISIS needs the HP. We did this on the system here and found we have far less problems. With computers we always expect a few crashes.
Luc Harmsen Anaspec, South Africa Technical support for E.M. operators, world wide. anaspec-at-icon.co.za TEL: ++ 27 (0) 11 476 3455 FAX: ++ 27 (0) 11 476 7290
-----Original Message-----
Hello to all,
We have been using a LEO 435 VPi SEM with an Oxford Inst. ISIS 300 integrated system since December of 1996. Since day one, we have had a series of problems with this integrated system, apparently caused by the common operating systems (first Win 3.1, then WIN95). It appears that the two systems do not like being integrated under one computer, as the sharing of resources seems to cause lurking problems that randomly appear, and cause system failures. I would be interested if any other users of the ISIS 300 system have had any other similar problems. Oxford claims that we are alone with our troubles, but I find this very hard to believe.
The first incarnation of our system used WIN 3.1 as the operating environment. This seemed optimum for the LEO as we encountered no system problems here. The ISIS though, would constantly run out of system resources and crash, usually without warning and lock up the system. It turned out that the ISIS is a resource hog (perhaps due to being written in Visual Basic), and the upgrade to ver 3.2 did not help at all. We then upgraded the system to Win 95, and the resource problem with the ISIS appears cured, but we have uncorked a beast in the LEO. The LEO is being upgraded to a full win95 system soon, but I am seeing more and more bugs with the ISIS.
Am I alone in believing the ISIS to be a poorly conceived and executed system? We have had endless conflicts with the operating system, to the point of having to reinstall win95 several times after apparent simple crashes, which seem to render several ISIS modules unavailable after that. The user interface appears haphazard from module to module, with no user definable defaults to customize operation to our preferences. The Labbook system forces users to collect data according to the x-ray system parameters. We like to structure our data by customer and job parameters, and keep all x-ray and microscope data under one directory. This is impossible under the ISIS Labbook system.
I am greatly disappointed with the integration of these two products. The LEO has been a fine system, with the operating environment keyed to usability, and endlessly customizable by the user. The ISIS is user unfriendly, and rigid in structure. Unless the user follows the Labbook protocols exactly, the system is unavailable to them.
Has anyone heard of a replacement operating environment for the Oxford system? Our complaints to Oxford have been dismissed, and LEO is unable to effect changes to the abysmal Oxford operating system. I wonder if I am alone in my frustration with this unfriendly and quixotic x-ray system.
Mike Warfield Hughes Space & Communications M&P Laboratory
At 02:45 PM 3/31/98 -0800, Paula Sicurello wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What you are looking for is a "focusing screen". It can be made following one of a number of materials, ranging from frosted glass to fresnel lenses to just a translucent piece of plastic --- anything which will allow you to catch the aerial image formed by the microscope. You can usually buy a piece of fresnel material at a science shop (most of the local malls have them now) or from a photographic store (I am not sure that they have one big enough to fit your Polaroid back. In any event, remove your film cassette and mount the screen in the plane where the film would be. If the adapter holding the camera system is adjustable, raise or lower the camera system until the image is in focus on the screen. (For more critical focus, try using either an 8x loop, available at any photographic store, or a simple 5x eyepiece from a light microscope). Once you have found this focal plane, lock the camera in position. You have now made your microscope and your camera system parfocal.
Being a light/confocal microscopist, I am unfamiliar with EM optics and how the image is formed at the film plane. In a light microscope, the depth of focus at the image plane is directly related to the magnification squared. As a result, using a low power objective gives the shallowest and most accurate focus in the image formed at the film plane and guarantees that all higher mags will automatically be in focus. }
Hope this helps.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
Garry, I, too, use the Paragon stain, but I do get a polychrome effect. I was wondering if you use a saturated Sodium Borate solution to enhance your staining. After drying the section on the hotplate, I put a 3 to 4 drops of Paragon stain and one drop of sodium borate on the slide (still on the hotplate). The combination of heat and sodium borate enhance the Paragon stain quite nicely. You might want to try this before adding H & E to your "to do" list.
Sharron G. Chism HT (ASCP) Electron Micropscopy Lab Harris Methodist Fort Worth Fort Worth, Texas ----------
In our diagnostic EM lab we currently use Paragon stain to stain our epoxy semi-thin sections, but it would be very nice if we had some kind of rapid H&E like stain that could give that kind of polychrome differentiation between the nucleus and the cytoplasm.
The Paragon stain is supposed to be a polychrome stain, but what we in effect see is more of a monochrome direct stain.
Does anyone know of a quick yet superior stain that might give us better results here?
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Dear Listers!
We have the problems with our JEM-4000EX: independent shut down after 10-30 minute of work. The possible cause is disturbance of logical schemes work of vacuum system. Does anybody else have the same problem, but more importantly has any one solved this problem ? Does anyone know how to make testing of vacuum system by CRT of microscope: how to load programme, where is check points and switchs e t.c.? I am not currently part of the list, so please mail to me directly.
Anton
gut-at-thermo.isp.nsc.ru
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We have a Durst Laborator 138 S enlarger. The lamp recently blew and we don't have a backup. We have tried to find a source for the special bulb but with no luck. Is Durst still in business? We have tried the Internet. The blown bulb is not the one that belongs in the enlarger. Thanks for any leads on Durst.
Researcher in the area of "Electron microscopy in plant research". Reference number 1716/98-4599.
At the Electron Microscopy Unit, Department of Plant Breeding Research, The Swedish University of Agricultural Sciences, Sval=F6v/Alnarp, Sweden. The EM Unit will be a common resource for the university's departments in Southern Sweden.
The candidate must have a doctor's degree, primarily not earlier than five years ago. Experience of work with plant material is a merit. Of importance are the scientific, pedagogic, adminstrative and other capacities of relevance for this occupation. Of importance also is the capacity to inform about scientific research in a popular way.=20
The appointment is first for two years, followed by another period of two years and the possibility for a further prolongation. The salary is individually determined (at least 20 000 Swedish crowns a month, before taxes). Women are engouraged to apply.
Together with the application, a short account on the applicant's scientific and pedagogic activities should be given. The application marked with the above-mentioned reference number, together with a certified C.V., merit and publication lists, and other documents (including scientific and pedagogic works) should be sent in two copies so that they reach the "Registrator, SLU, Box 7070, S-750 07 Uppsala, Sweden" at the latest on April 23, 1998.
For further information contact professor Waheeb Heneen, tel +46 418 667064 or fax +46 418 667081.
I work for VayTek, and would like to respond to Alex Black's question on 3D reconstruction using TEM.
Yes it's possible. You must aquire a series of images in sequence and equidistant from each other. They must be all the same size, resolution, and magnification.
I'm aware that this is a tall order. I've just completed doing a 3D reconstruction of some TEM images with great results. The process involved some very tricky TEM work, and some sofisticated image processing using three software packages.
I'm not at liberty to give you more details on the TEM work, but if you're interested in knowing more about the process, please e-mail me directly. I'll contact the lab doing this and ask them to contact you. I'll also fill you in on the software involved.
Best regards
Patrick Guerin Customer Technical Support Engineer VayTek, Inc. 305 West Lowe Avenue, Suite 109 PO Box 732 Fairfield Iowa 52556-0732 Tel : 1-515-472-2227 ext. 103 Fax : 1-515-472-8131 WWW.VAYTEK.COM E-mail : pguerin-at-vaytek.com
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Dear Sara, } } Does anyone have experience growing tissue culture cells on gold grids } with support films? We want to do IEM of filaments after digestion of } the cytoplasmic membrane, } The one time I tried this (and succeeded) I just spread the carbon- formvar-coated gold grids on the bottom of a culture dish, added PTK1 cells and let the cultures sit at 37 degrees. YMMV, if your cells do not grow over carbon-formvar; however, if you grow your cells on a treated culture dish to promote adhesion, treating the carbon-formvar film the same way should work (IMVVHO).
} Articles describing this technique mention specific kinds of reagents, } grids, etc. Are there better brands, methods? We'd appreciate any tips } to avoid a lot of trial and error. } } Source of gold grids? (supplier)
Probably makes no difference unless the "gold" in the grids contains different impurities, and if these affect cell growth.
} Type of support film? Formvar/collodion)
If the hydrophobicity, texture, etc. of the film matches that of the usual culture surface, you are likely to get the best results.
} Carbon coat? (heavy/light)
I'd reccommend it, unless you need uncoated formvar to match adhesion conditions.
} Method of sterilization? (UV/gas)
Probably irrelevant, unless the surface is affected adversely. Bear in mind that I am most definitely not an expert. Good luck. Yours, Bill Tivol
I know we should drop this subject but...........This is like dejavu. Several years back I was teaching the microscope structure/function section of an EM course and began thinking how to explain the theory of resolution, trying to make it interesting and not too complicated. My mind wandered (as it does quite often these days) and got to thinking about the effects of space and the resolution potential of an electron microscope, The Phillips engineer happened to be here and blew it past him. He wasn't sure what effects it would have but thought it interesting. He said the main limitation in current scopes was the quality of the lenses, but could we make better lenses in space then add that to the vacuum, cold, and magnetic/gravitational variable and get a significant increase in resolution? Just a thought, I realy don't want us to get into the physics of this.
Simon C. Watkins Ph.D. Associate Professor Director, Center for Biologic Imaging University of Pittsburgh Pittburgh PA 15261 tel:412-648-3051 fax:412-648-2004 URL: http://sbic6.sbic.pitt.edu
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As a class project, I am writing a proposal for a (fictitious) SEM lab for a university geosciences department. I am attempting to obtain the Alderson book on EM lab design, but would like additional references. Any citations would be appreciated.
Also, if any geoscientists have any suggestions for how they would like to see an SEM/EDS lab equipped, I would be happy to hear them.
with TCP; Thu, 02 Apr 98 17:04:05 EST Received: from [137.99.40.57] by oracle.pnb.uconn.edu with SMTP (Eudora Internet Mail Server 1.2); Thu, 2 Apr 1998 17:12:29 -0500 Message-Id: {v01510109b149acd24f9d-at-[137.99.40.57]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I have noticed that in EM immunogold-labelling protocols, BSA is often used in buffer as an initial blocking step. In some protocols a BSA buffer rinse is also used before the secondary antibody, and/or is added to the primary and secondary antibody solutions. In other cases it is omitted. My question, to immuno experts on the listserver, is whether there is a DISadvantage to adding the BSA after the initial blocking step. As I understand it BSA reduces non-specific binding, so it would seem prudent to add it whenever possible.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
In the mid-1970's NASA did send to universities a request for suggestions for experiments to put into space. At the time I was at Bristol and put it to my colleagues that would should submit a proposal to put a TEM into space. My argument was that all the instability problems would go away. No vibrations from vacuum pumps because there are no vacuum pumps - no vibration from anything else either. No instabilities in the lens coils because they would all be superconducting with no need for cooling equipment. No sample contamination because the vacuum is clean. It would make for perfect images. I was motivated by the thought that I might get to go into space to operate it (I was younger then). I could not persuade my colleagues to take me seriously. Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
You wrote: I have noticed that in EM immunogold-labelling protocols, BSA is often used in buffer as an initial blocking step.
If BSA is a problem i.e. failing to block non-specificity , there are several other blockers that can be used. Ovalbumin and fish gelatin are often "cleaner" blocking agents for gold labeling procedures. Also the purity of BSA can be a factor, I always buy a grade of BSA that contains greater than 95% BSA of MW 66,000. If I can be of further help, feel free to contact me off the listserver. Marge
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
We are looking at hi-res CCD cameras, preferably with video-rate output as well, for 300kV TEMS (1000 x 1000 pixel, 12 bit minimum), and would appreciate the benefit of other's experience regarding quality, ease of use. longevity, radiation problems, value-for -money, etc. TIA Sally Stowe ---------------------------------------------------------------------- Dr Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: Box 475 ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 (0)2 6249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, Australia 2601 http://online.anu.edu.au/EMU/home.htm |AUSTRALIA 0200
I have noticed that in EM immunogold-labelling protocols, BSA is often used in buffer as an initial blocking step. In some protocols a BSA buffer rinse is also used before the secondary antibody, and/or is added to the primary and secondary antibody solutions. In other cases it is omitted. My question, to immuno experts on the listserver, is whether there is a DISadvantage to adding the BSA after the initial blocking step. As I understand it BSA reduces non-specific binding, so it would seem prudent to add it whenever possible.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
I always use it in all solutions where antibodies and/or colloidal gold are used, and as an initial blocking solution. I don't know of any reason not to use it to reduce nonspecific staining, although cost might become an issue if you do your washes in between steps in very large volumes.
At 05:01 PM 4/2/98 -0400, MARIE CANTINO wrote:
} I have noticed that in EM immunogold-labelling protocols, BSA is often used } in buffer as an initial blocking step. In some protocols a BSA buffer } rinse is also used before the secondary antibody, and/or is added to the } primary and secondary antibody solutions. In other cases it is omitted. } My question, to immuno experts on the listserver, is whether there is a } DISadvantage to adding the BSA after the initial blocking step. As I } understand it BSA reduces non-specific binding, so it would seem prudent to } add it whenever possible. } } Marie } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology, U-131 } University of Connecticut } Storrs, CT 06269 } Ph: 860-486-3588 } Fax: 860-486-1936 } } } } } _____________________________________ Craig Lending SUNY Brockport Department of Biological Sciences Brockport, NY 14420
On Fri, 3 Apr 1998, SALLY STOWE wrote: } We are looking at hi-res CCD cameras, preferably with video-rate } output as well, for 300kV TEMS (1000 x 1000 pixel, 12 bit minimum), } and would appreciate the benefit of other's experience regarding } quality, ease of use. longevity, radiation problems, value-for } -money, etc. } Sally, I am also interested in this information, please forward anything not posted to the list. Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Dear Marie, I think the protocol you need to use depends on your primary antibody. If BSA fails to block aspecific binding, skimmed milk powder can be a better blocking agent. But whatever blocker I use, I include it in all the reaction steps. Armelle
Armelle Gollotte Biotechnology Department Scottish Agricultural College West Mains Road Edinburgh EH9 3JG Tel: (44) 131 667 1041 Fax: (44) 131 667 2601 a.gollotte-at-ed.sac.ac.uk
} If BSA is a problem i.e. failing to block non-specificity , there are } several other blockers that can be used. Ovalbumin and fish gelatin are } often "cleaner" blocking agents for gold labeling procedures. Also the } purity of BSA can be a factor, I always buy a grade of BSA that contains } greater than 95% BSA of MW 66,000.
In order to get a very 'clean' blocking I always use modified BSA-C (Aurion) It is more expensive than quality grade BSA and Cold Fish gelatin but I find it is much easier to use (you get it as a non-sticky solution) and it gives a very clean and efficient block. I have never had any problems using modified BSA-C.
Do any of you have benchmarks which you use when evaluating cameras, digitizing boards, or image analysis systems? Also, are you using any type of standardized slides for comparing systems?
Thanks
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
Do any of you have benchmarks which you use when evaluating cameras, digitizing boards, or image analysis systems? Also, are you using any type of standardized slides for comparing systems?
Thanks
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
At 09:18 AM 4/2/98 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
George,
Durst headquarters is at 10 County Line Road, Suite 29, Branchburg, NJ 08876. Phone is (800) 463-8778, and their fax is (908) 429-0777. You can find a web site for them at http://www.photoshopper.com/durst/index.html.
If this fails to help in your bulb search, try such distributors as Bulbman (sorry, don't have their address handy) or another of the specialists in lighting equipment and bulbs. They have catalogs available and the one's I've dealt with are very willing to track down obscure bulb types if you can provide them with descriptions, identifying numbers, applications, etc.
Good luck.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
First of all , I am no immuno expert. I use ovalbumin for initial blocking and wash grids on porcelain spot plat filled with buffer and use blocking step again before secondary antibody reaction. I see no merit of adding BSA or any other blocking agent during washing.
I have noticed that in EM immunogold-labelling protocols, BSA is often used in buffer as an initial blocking step. In some protocols a BSA buffer rinse is also used before the secondary antibody, and/or is added to the primary and secondary antibody solutions. In other cases it is omitted. My question, to immuno experts on the listserver, is whether there is a DISadvantage to adding the BSA after the initial blocking step. As I understand it BSA reduces non-specific binding, so it would seem prudent to add it whenever possible.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
I am preparing cross-sectional TEM samples of sapphire. I am trying to find a good way to measure the samples' thickness when I am under about 70 micron. Any ideas?
For ICC blocking I use gelatin (Sigma Gelatin, G-9382, from bovine skin, type III, approx. 225 bloom). This preference results from a sabbatical year (1985-86) I spent in the lab of Daniel Louvard, then at the Pasteur Institute in Paris. They had tried a good many blockers and decided on this gelatin (easy to obtain and use, few cross reactions, etc.). They blocked with 2% gelatin in PBS, and thereafter maintained 0.2% in washes and vehicle for primary/secondary antibodies. A 10% gelatin stock was stored in the refrigerator, and was warmed (for example under hot tap water) to liquify for use.
Kent
A. Kent Christensen Department of Anatomy and Cell Biology Medical Sciences II Building University of Michigan Medical School Ann Arbor, MI 48109-0616 akc-at-umich.edu Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
--------------------------------------
} } } } MARIE CANTINO {CANTINO-at-ORACLE.PNB.UCONN.EDU} 04/02/98 } 04:01pm } } } } } I have noticed that in EM immunogold-labelling protocols, BSA is often } used } in buffer as an initial blocking step. In some protocols a BSA buffer } rinse is also used before the secondary antibody, and/or is added to the } primary and secondary antibody solutions. In other cases it is omitted. } My question, to immuno experts on the listserver, is whether there is a } DISadvantage to adding the BSA after the initial blocking step. As I } understand it BSA reduces non-specific binding, so it would seem } prudent to } add it whenever possible. } } Marie } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology, U-131 } University of Connecticut } Storrs, CT 06269 } Ph: 860-486-3588 } Fax: 860-486-1936
What is Paragon stain, and where can I get it? It sounds like it's exactly what I need, and I'm sure it would be usefulto the rest of the list if it's as wonderful as it sounds. TIA. Lesley Weston.
On Thu, 2 Apr 1998, Chism, Sharron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Garry, } I, too, use the Paragon stain, but I do get a polychrome effect. } I was wondering if you use a saturated Sodium Borate solution to enhance } your staining. After drying the section on the hotplate, I put a 3 to 4 } drops of Paragon stain and one drop of sodium borate on the slide (still } on the hotplate). The combination of heat and sodium borate enhance the } Paragon stain quite nicely. You might want to try this before adding H } & E to your "to do" list. } } Sharron G. Chism HT (ASCP) } Electron Micropscopy Lab } Harris Methodist Fort Worth } Fort Worth, Texas } ---------- } From: Garry Burgess } To: 'Microscopy' } Subject: H&E Like Stain for Epon/Araldite } Date: Wednesday, April 01, 1998 2:25PM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } In our diagnostic EM lab we currently use Paragon stain to stain our } epoxy semi-thin sections, but it would be very nice if we had some kind } of rapid H&E like stain that could give that kind of polychrome } differentiation between the nucleus and the cytoplasm. } } The Paragon stain is supposed to be a polychrome stain, but what we in } effect see is more of a monochrome direct stain. } } Does anyone know of a quick yet superior stain that might give us better } results here? } } } Garry }
In a counterpoint to Mike Warfield's message, we have been very happy customers of Oxford (Link Systems in its previous incarnation) since 1985.
We have one integrated and one stand alone ISIS systems. The stand alone is a 2 y.o. ISIS 3.00 EDS replacement of an obsolete Noran on a 16 y.o. AMRAY 1600 SEM. The other unit is a brand new ISIS 3.2 integrated into an AMRAY 3200/C ECO SEM. (We also operate a 1985 vintage AN10 on a VG STEM from the era.)
I was intimately involved in evaluation and purchase of both ISIS systems. During the last five years, we have evaluated in exhausting detail every brand of EDS equipment and found the Oxford detectors to offer the best spectral resolution, and the software to be the most comprehensive and useful. The Oxford software worked flawlessly in all demos and in our lab. under Windows 3.1, 95 and NT 4.0.
We are an industrial lab serving several of our plants and many of the plants' customers. A high throughput, very high reliability, and good rapid service on our analytical equipment are paramount. In 1997, our old SEM was used to examine over 2,100 samples, each a unique specimen, and more than 90% required EDS analysis. During that time, the ISIS 3.00 performed very well and operated without a mishap.
As is the case in any new installation, we ran into initial problems, but Oxford has bent over backwards to help us out through both installations.
In the 1996 Noran upgrade, we purchased a used (demo) ISIS 3.00 analyzer sans the detector to inexpensively replace a third party kludge fix up of an obsolete Noran EDS system, but we were keeping the old detector as it seemed to work just fine. Following the installation, transient high noise levels were experienced in the EDS spectra. Over the next few weeks, Oxford engineers replaced every board on the system and, suspecting an incompatibility with the modified Noran detector, Oxford provided us gratis a brand new 30 mm2 thin window detector with 126 eV peak resolution. In the end, it turned out that an intermittent noise originated from a flaky pulse processor power supply, but Oxford let us keep the $20,000+ detector; well above and beyond the call of duty. Since the replacement of the power supply in February 1996, the system has operated flawlessly and it easily adapted to an upgrade from Windows 3.1 to 95.
In the ISIS 3.2 integrated installation on the AMRAY 3200/C ECO SEM, we ran into a problem of insufficient x-ray count rate. The detector resolution was just fine (118 eV), but we needed to acquire spectra for 200 seconds with a higher beam current for counts comparable to 50 s acquisition on the old SEM. It turns out that I goofed and specified a 10 mm2 detector for an operator used to working with 30 mm2. Also, the Oxford detector is aligned for a sweet spot at a 25 mm working distance (from the objective polepiece to the sample plane) and a tightly focused collimator prevented us from getting x-rays at working distance shorter than 20 mm or greater than 30. To compound the problem, an interference from a third party add-on, a Robinson back-scatter detector, required the nosepiece of the Oxford EDS detector to be withdrawn above the bottom of the objective polepiece when the Robinson was inserted. The combination of smaller detector area and a huge detector to sample distance killed the x-ray count rate. When we contacted Oxford with our dissatisfaction of the system performance, the response was initially slow and unfocused until we got past the newly expanded inexperienced service staff once it became clear that our problems went beyond the standard garden variety. An experienced service engineer flew in from outside his working area within 10 days of our initial call, and with an aid of a knowledgeable sales rep., helped to fully diagnose the problem. The collimator on the detector nose piece has been modified and we have tested it to our satisfaction at the sample to objective pole piece working distance range of 15 to 40 mm. Oxford has also promised to make in a short order a new EDS detector mounting flange to remove the interference between the EDS and the Robinson backscatter detectors.
Having been there, I empathize with Mike Warfield's frustrations with his equipment not working as well as it should. However, I think that not the full story has been exposed here and that the finger pointing at Oxford is less than fair.
Point #1:
Our older ISIS 3.00 runs just fine on a 66 MHz HP Vectra with 16 Mbytes RAM. It is a stand alone system and has had no computer related problems at all under Windows 3.1 nor 95. (We upgraded to 95 solely for better compatibility with our LAN.) The integrated ISIS 3.2 and AMRAY 3200/C run on a Micron 133 MHz P5 with 32 Mbytes RAM and 512 Kbytes L2 Cache under Windows NT4.0. No computer related problems have manifested themselves in this system either, though perhaps surprisingly the old system is somewhat more responsive. (I do wish we did succeeded in getting through to the SEM vendor that an integrated system requires a 300 MHz P5. Why pinch a few hundred dollars on a $300,000+ system?)
It occurs to a few users, and it seems to fewer SEM vendors, but when two machines (as in an integrated SEM & EDS system) are squeezed onto one computer, at least twice as much RAM and twice as high CPU frequency are required, and the perforce will still not be as good as of two stand alone systems. That is because, though your CPU may be screaming at 500 MHz, the RAM and the system buss still chug along at the same old 33 MHz while two suites of software are time-slice multitasked and interrupts from two sets of hardware are simultaneously serviced. Nevertheless, what little one may have to give up in responsiveness WHEN AN APPROPRIATELY SCALED COMPUTER is used for an integrated system is more than made up in convenience. (I just can't imagine simultaneously operating two keyboards, two pointing devices, and watching three monitors with any efficiency.)
Mike does not list the specifications of his PC, but I wonder whether it's a poky old 486 with 16 Mbytes of RAM which works just fine running either the SEM alone or the EDS alone but bogs down when asked to do both simultaneously. (If there is not enough RAM the system may be rally grinding to a halt while sloshing the programs back and forth between RAM and the "virtual RAM memory" on the hard drive.) A RAM boost or a PC upgrade might help here.
Note that this is an integrated EDS system onto the SEM by the SEM manufacturer, and the responsibility for sufficient computer resources for both systems lie with LEO and not with Oxford.
Point #2:
Windows 95 is a terrible operating system in general, and especially for real time multitasking. It does not provide sufficient system resources and it does not clean up after itself (just check the number of *.TMP files accumulated on your hard drive). If fixes suggested in Point #1 don't work, upgrade from 95 to NT4. If the software runs under 95 chances are good it will run under NT4. ISIS 3.2 works flawlessly on our integrated machine under NT4.
Again, the responsibility for choosing the appropriate operating system lies with LEO.
Point #3:
Some SEM manufacturers rely on the purchaser to choose the EDS system and will perform the integration as a courtesy to the purchaser. However, the satisfactory site installation and subsequent service contract on the EDS unit is the responsibility of the EDS manufacturer. This is the case with our integrated system.
Other SEM manufacturers purchase an EDS unit as a commodity and sell the SEM with an integrated EDS as one package. I imagine there is then just a single service contract and the SEM vendor is supposed to service both the SEM and the EDS. In principle this is a better deal because there should be less finger pointing, but in reality the SEM vendor's tech. support is not up to snuff on the EDS end and the user pays in frustration. I think the LEO integration falls in this second category.
Point #4:
I am surprised that Mike is expressing his dislike of the Oxford user interface at this point in the game. I think a more appropriate time frame would have been during the evaluation period, well before the whole system was purchased and paid for.
These opinions are mine alone and in no way reflect the views of my employer. I bear no interest in AMRAY, OXFORD, or LEO.
Valdemar Furdanowicz
valdemar-at-fast.net Homer Research Laboratories Bethlehem Steel Corporation
---------- } From: mdwarfield-at-mail.hac.com } To: Microscopy-at-sparc5.microscopy.com } Subject: LEO 435 VPi with ISIS 300 X-Ray Analysis System } Date: Wednesday, April 01, 1998 9:38 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello to all, } } We have been using a LEO 435 VPi SEM with an Oxford Inst. ISIS 300 } integrated system since December of 1996. Since day one, we have had a } series of problems with this integrated system, apparently caused by } the common operating systems (first Win 3.1, then WIN95). It appears } that the two systems do not like being integrated under one computer, } as the sharing of resources seems to cause lurking problems that } randomly appear, and cause system failures. I would be interested if } any other users of the ISIS 300 system have had any other similar } problems. Oxford claims that we are alone with our troubles, but I } find this very hard to believe. } } The first incarnation of our system used WIN 3.1 as the operating } environment. This seemed optimum for the LEO as we encountered no } system problems here. The ISIS though, would constantly run out of } system resources and crash, usually without warning and lock up the } system. It turned out that the ISIS is a resource hog (perhaps due to } being written in Visual Basic), and the upgrade to ver 3.2 did not } help at all. We then upgraded the system to Win 95, and the resource } problem with the ISIS appears cured, but we have uncorked a beast in } the LEO. The LEO is being upgraded to a full win95 system soon, but I } am seeing more and more bugs with the ISIS. } } Am I alone in believing the ISIS to be a poorly conceived and executed } system? We have had endless conflicts with the operating system, to } the point of having to reinstall win95 several times after apparent } simple crashes, which seem to render several ISIS modules unavailable } after that. The user interface appears haphazard from module to } module, with no user definable defaults to customize operation to our } preferences. The Labbook system forces users to collect data according } to the x-ray system parameters. We like to structure our data by } customer and job parameters, and keep all x-ray and microscope data } under one directory. This is impossible under the ISIS Labbook system. } } I am greatly disappointed with the integration of these two products. } The LEO has been a fine system, with the operating environment keyed } to usability, and endlessly customizable by the user. The ISIS is user } unfriendly, and rigid in structure. Unless the user follows the } Labbook protocols exactly, the system is unavailable to them. } } Has anyone heard of a replacement operating environment for the Oxford } system? Our complaints to Oxford have been dismissed, and LEO is } unable to effect changes to the abysmal Oxford operating system. I } wonder if I am alone in my frustration with this unfriendly and } quixotic x-ray system. } } Mike Warfield } Hughes Space & Communications } M&P Laboratory } }
I also have a Durst 138 S enlarger. I use both the Pullam light source (point)or 110V-200 Watt opale lamp (basically a giant light bulb). Durst is still in business. I called the supplier I use and they can order it from Durst for you. They are: Espinoza Photo Systems, 5213 A Clayton Road, Suisun, CA 94585 Phone (707) 428-0912. Ask for Patty. JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94022
At 05:08 PM 3/31/98 -0500, Heeschen, Bill (WA) wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Call the folks at Zygo. I am not sure if their complete software is available, but they may have a fragment available which would solve your problem nicely. Try either Bob Smythe at (860)704-5101 or Les Dock (860)347-8506 (general number).
Wyko Corp. also has extensive experience in this area. They were recently acquired by Veeco. The last number I have for them is (602)741-1297.
Best of luck.
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
Hi Simon, We've been buying ITO glass from Donnelly Optics Corporation of Tucson, Arizona at 1-888-250-4455 (or 520-806-3800, Sales dept.). Although our application is different we have been using 2 inch squares, the ITO having a conductivity/resistivity of about 100 ohms per inch. If you need only a few pieces of this glass I could arrange to get some to you, otherwise try contacting Donnelly.
Good Luck, Paul Gerroir ----------
Folks, do any of you know of a supplier for ITO glass for making heated culture chambers Thanks Simon
----------------------------------------------------------------------- Simon C. Watkins Ph.D. Associate Professor Director, Center for Biologic Imaging University of Pittsburgh Pittburgh PA 15261 tel:412-648-3051 fax:412-648-2004 URL: http://sbic6.sbic.pitt.edu
and add a few grains of Thymol to preserve it. LET IT AGE 2 WEEKS BEFORE USING!
} What is Paragon stain, and where can I get it? It sounds like it's exactly } what } I need, and I'm sure it would be usefulto the rest of the list if it's as } wonderful as it sounds. TIA. } Lesley Weston. } } } } On Thu, 2 Apr 1998, Chism, Sharron wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Garry, } } I, too, use the Paragon stain, but I do get a polychrome effect. } } I was wondering if you use a saturated Sodium Borate solution to enhance } } your staining. After drying the section on the hotplate, I put a 3 to 4 } } drops of Paragon stain and one drop of sodium borate on the slide (still } } on the hotplate). The combination of heat and sodium borate enhance the } } Paragon stain quite nicely. You might want to try this before adding H } } & E to your "to do" list. } } } } Sharron G. Chism HT (ASCP) } } Electron Micropscopy Lab } } Harris Methodist Fort Worth } } Fort Worth, Texas } } ---------- } } From: Garry Burgess } } To: 'Microscopy' } } Subject: H&E Like Stain for Epon/Araldite } } Date: Wednesday, April 01, 1998 2:25PM } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } In our diagnostic EM lab we currently use Paragon stain to stain our } } epoxy semi-thin sections, but it would be very nice if we had some kind } } of rapid H&E like stain that could give that kind of polychrome } } differentiation between the nucleus and the cytoplasm. } } } } The Paragon stain is supposed to be a polychrome stain, but what we in } } effect see is more of a monochrome direct stain. } } } } Does anyone know of a quick yet superior stain that might give us better } } results here? } } } } } Garry } } } }
I am faced with a problem of selectively staining teflon particles impregnated in nickel matrix. I would really appreciate any suggestions regarding the chemicals and/or the procedure to stain teflon particles.
We have made samples of sapphire using the wedge technique and it works = very well. Sapphire is a hard material, therefore, it is easy to control = how much material you are removing even though it takes longer to thin = it down. Since sapphire is transparent to light we glue a piece of = silicon to the sample and use the change of color of the Si piece as a = reference on how thin the sapphire sample is. =20
Microscopists: There is an actual H&E like stain. Use Celestin Blue B and Eosin after removing the plastic. A stain like this is almost indistinguishable from H&E and in fact microtomists may all be using Celestin blue B as the supplies of tropical logwood (source of dye for hematoxylin) dry up.
There is a lot that can be said about additives to incubation buffers, the composition of blocking media and washing procedures (and a lot more to come probably!). I don't want to clog everyone's mailbox: we offer two Newsletters (free) with in-depth information on the topic of specificity and non-specific results in immuno. Those who are interested are welcome to request Newsletters through our website http://www.aurion.nl. There is a number of FAQ's re. immunolabeling taken up as well.
Just in short: A distinction should be made between a blocking buffer and an incubation buffer. They serve different purposes: the blocking buffer serves to bind protein (BSA, Gelatine, Ovalbumine, Casein etc.) to sticky (hydrophobic) specimen areas. The incubation buffer and wash buffer should prevent background based on hydrophilic interactions (charges) by competition. Different additives may be in order. Many additives may work, although empirical finding sooner than logics seems to be at the basis of their application. An additive that seems to work fine with one set of primary antibody and secondary reagent seems to be less suited with different reagents. With this in mind we have developed Aurion BSA-c, which proves to be a great help with all kinds of antibodies and immunoreagents. } From a cost point of view it may be tempting to leave out additives during the washing steps, although during washing unreacted antibodies or labels may stick to the specimen when they're no longer prevented to.
We use poly-l-lysine coated slides for all our in situs. During the course of a month we will go through several dozen slides. Consequently we have to coat slides quite often. All attempts with commercially available slides have met with unsatisfactory results. We prefer the quality of slides we generate "in house". What I would like is some advice on storage of a large vol. of poly-l-lysine solution that can be accessed and used on a regular basis. We make up 4 L of .05 mg/ml poly-l-lysine solution in 10 mM Tris buffer in a large tray, store this is the freezer and thaw when need. This system allows us to coat several dozen slide simultaneously (we soak geletin coated slides in the PLL soln. for 10-30 min). The problem is that the solution appears to generate a ppt. during storage, a white "fluff".
Looking for all kinds of ideas
Thanks, Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
A friend gave my your e-mail. This is probably old news, however on the off chance this will help, here is an address. CH-Imports Ltd 3410 Deep Green Drive Greensboro NC 27401 Phone 910-282-9734 Fax 910- 288-3375 Their Catalog shows 3 items: Cedarwood Atlas Mardoc OG 71.50/KG Cedarwood Crude Kenya OG 41 /kg Cedarwood Forte USA OG 64.40 /kg Cedawood Virginia OG 33.50 /kg OG stands for Organically grown.? Larry Albright
Heaven is where: the police are British, the cooks are French, the mechanics are German, the lovers are Italian, and all is organized by the Swiss.
Hell is where the police are German, the cooks are British, the mechanics are French, the lovers are Swiss, and all is organized by the Italians. albrite-at-Plasma-Art.com 419 Sunset Avenue Venice CA 90291 310-399-0865 310-392-9222 FAX
"Fundamentals and Applications of Light Microscopy"
July 27-31, 1998 Waltham Massachusetts
A 5-day practical course on how to get maximum information from the light microscope, taught by experienced microscopy problem-solvers.
Hands-on experience, a firm foundation in the principles of LM, and knowledge of contrast enhancement techniques will be emphasized. A full range of reflected and transmitted light microscope and contrast equipment will be available for use by the students.
Although ideal for beginners, the course will also benefit experienced workers. Students are encouraged to bring their own samples.
For Further information contac: Mary McCann McCann Imaging 161 Claflin Street Belmont MA, 02178 Telephone:617-484-7865 Fax: 617-484-2490 E-mail address: mccanns-at-tiac.net URL: www.microscopyed.com
On Fri, 3 Apr 1998 scm5-at-ix.netcom.com (Sanjay Mehta) wrote:
} I am faced with a problem of selectively staining teflon particles } impregnated in nickel matrix. I would really appreciate any suggestions } regarding the chemicals and/or the procedure to stain teflon particles.
We have sometimes looked at PTFE under the electron microscope, with some of our preparations involving "Tetra-Etch", which is, I think, a solution of sodoum napthalene compound in an organic solvent (perhaps tetralin OR tetrahydrofuran, hence the name.) This turns the surface of the PTFE into a sort of carbon, which might take heavy metal stain (but we were using replication, so we did not stain).
If I knew what sort of method was required (SEM, TEM, etc.) I might be able to be more specific.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Dear Neil, would you, please, kindly provide us with the information, what sections = should be stained (paraffin, celloidin, if resin(s) which kind of??). = Either, I think it would be quite difficult to stain only ECM which = normally would be divided into pericellular matrix, territorial matrix = and interterritorial matrix, respectively ("chondrones"). If you're looking for "cartilaginous lacunae" in order to measure such = structures: believe me: they are artifacts of specimen preparation due = to wrong fixation/dehydration schedules......but, perhaps, your = colleague would be doing those measurements for a scientific evaluation = of artifact-induction by poor fix, dehydration, embedding criterions. If you like we could discuss that matter off the list.....
best wishes and good luck Wolfgang
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: Neil Hand[SMTP:mpznhand-at-unix.ccc.nottingham.ac.uk] Gesendet: Montag, 06. April 1998 06:49 An: HistoNet-at-Pathology.swmed.edu Betreff: Q: Cartilage staining, specific
This is a inquiry on behalf of a colleague (not working in Nottingham) I recently received.
"I am currently looking for an optimal staining or sectioning procedure = for samples of ear and articular cartilage, whereby the stain is confined to the extracellular matrix without staining (or very light staining) the cells and nuclei within the cartilaginous lacunae. The reason for this = is to be able to accurately assess the ratio of extracellular matrix to lacunar space by computerized morphometric analysis. Currently, unavoidable staining of cells is obscuring the measurements made by the = morphometric program, which uses gray scales and contrast to arrive at = values".
Further details I do not have. Any suggestions you may have on this = matter would be appreciated.
Neil Hand, Nottingham, England UK.
___________________________________________________________________ Neil Hand Department Histopathology, University Hospital, Nottingham NG7 2UH. work : Tel: (0115) 924 9924 extension 43725 ___________________________________________________________________
For everyone interested, here is the procedure we use to stain semi-thin sections with Paragon stain. Paragon Stain: Ethyl Alcohol ................ 1,028 mls Deionized water .......... 2,397 mls These combine to make up a 30% ETOH solution. (Set this aside for the moment.)
Toluidine Blue ............... 25 grams Basic Fuchsin ................ 9.25 grams Combine these in a 1 gallon (or 4 liter) container.
Add the 30% ETOH and mix well.
Allow the stain to stand for several days, then filter into an amber gallon (4 liter) bottle to protect the stain from light. (It will last a LONG time!)
Filter the stain again before use.
Staining Procedure: Transfer cut section onto a drop of deionized water previously place on a clean microscope slide.
Place the slide holding the section on a hot plate set at 200 degrees C. Allow the water to evaporate and the section to dry.
Drop 3 to 4 drops of filtered Paragon stain on the dry section. Add a drop of saturated solution of Sodium Borate. Continue to heat for 10 - 15 seconds or until the stain begins to show a metallic sheen.
Using a wash bottle of 50% ETOH, gently wash off all the excess stain.
Dry the slide with gentle heat using a blow dryer ... DO NOT DRY ON HOT PLATE!
At this point, maybe I should point out that I use Spurr's resin. I also keep a little Spurr's in the freezer to use as a coverslipping medium for my semi-thin sections. I hope this is helpful!
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Fort Worth Fort Worth, Texas
In a message dated 98-04-02 17:21:53 EST, CANTINO-at-ORACLE.PNB.UCONN.EDU writes:
{ { I have noticed that in EM immunogold-labelling protocols, BSA is often used in buffer as an initial blocking step. In some protocols a BSA buffer rinse is also used before the secondary antibody, and/or is added to the primary and secondary antibody solutions. In other cases it is omitted. My question, to immuno experts on the listserver, is whether there is a DISadvantage to adding the BSA after the initial blocking step. As I understand it BSA reduces non-specific binding, so it would seem prudent to add it whenever possible.
Marie } } Marie,
I agree with you, and this is the way I've always used BSA. Some Abs seemed to give more problems than others, or perhaps it was the phase of the moon, I was never sure.
I would put it in all solutions (including the buffers and the diluents for the Ab and the gold), only removing it from the final buffer and water washes at the very end after the immunolabeling has been completed. This seemed to work for me, and my reasoning was always, "if it ain't broke...."
Best regards, Bob ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Cryostats / Microtomes / Tissue Processors / Embedding Centers / Slide Stainers / Glass Coverslippers / Microscopes (Representing Leica since 1967) / Fiber Optic Systems / Linear Measuring / Micromanipulation (Linear Encoded, Video) / Image Analysis, Archiving, Capture / Video / Video Printers (Cooled CCD, Digital, RGB, Super VHS, 3-chip) / Vibration Isolation Systems / Programmable Stages / Heating & Cooling Stages / Motion Control, Positioning and Measurement Systems
Dear Linda, } } What I would like is some } advice on storage of a large vol. of poly-l-lysine solution that can be } accessed and used on a regular basis. We make up 4 L of .05 mg/ml } poly-l-lysine solution in 10 mM Tris buffer in a large tray, store this is } the freezer and thaw when need. } The problem is that the solution appears to generate a } ppt. during storage, a white "fluff". } Have you looked at the "fluff" under the LM or EM? If it is poly- l-lysine, you may have to divide the PLL into aliquots before freezing & redissolve each aliquot before use. If it is something else--I can't im- aging from your description that it could be bacterial growth, but a brief examination would tell--you would have to identify it before deciding what to do. Good luck. Yours, Bill Tivol
} Is it possible for one week old paraformaldehyde (originally } in argon sealed ampules)-EM grade- to actually permeabilize membranes? } (by some glycol formation). All comments are welcome. } Are these membranes--such as plasma membranes, mitochondrial inner or outer, etc.--with which other microscopists have had preparative experience, or are they unusual in some respect? Since I might expect paraformaldehyde and/or some of its polymers to react with some membrane components, it is possible that the membranes could become permeable to some substances. What substance did they become permeable to? What was its molecular mass? Yours, Bill Tivol
Does anyone have a dysfunctional HummerVI sputter coater, which could be cannibalized or purchased as a surplus item. I need vacuum pump (built in, type G-50D made by ULVAC-Sinku Kiko or equivalent) for this coater. Thank you in advance for your reply.
Chris Terlecki
Applied Analytical Sciences 3303 Harbor Blvd. Ste. H-4 Costa Mesa, CA 92626
What are recommended conditions for staining Epon generic or Spurr resin sections with either the Celestin Blue/Eosin or Paragon stain? I would like to know desired concentrations, temperature, time, etc as starting points and any handy hints to improve success on a variety of tissues.
I have someone wanting to use a stain like this on rat liver, spleen and lung. He has looked at the material in parafin with H & E but wants to get better quality thick sections and then thin sections for TEM of selected areas.
Debby Sherman, manager Microscopy Center in Agriculture Purdue University
--------------------------------------
Microscopists: There is an actual H&E like stain. Use Celestin Blue B and Eosin after removing the plastic. A stain like this is almost indistinguishable from H&E and in fact microtomists may all be using Celestin blue B as the supplies of tropical logwood (source of dye for hematoxylin) dry up.
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Please remove us from the list amrayinc-at-aol.com
I measure the thickness of my samples that I have mechanically thinned two ways. 1) a dial indicator mounted on a granite check block. Mine is digital inches/mm with mm to the third decimal. Look in a tool catalog. Zero on a mounting stub and slide the sample underneath. 2) a dimpler. Zero the dimpler on a mounting stub and then place the sample without wax on a measure the thickness. ----------
I am preparing cross-sectional TEM samples of sapphire. I am trying to find a good way to measure the samples' thickness when I am under about 70 micron. Any ideas?
If the HummerIV is anything like the III I once owned, a direct replacement is not required. I managed to "shoehorn in" a Welch pump. If I had it to do over again, I wouln't even put the pump in the box, but behind it. ...Still a fairly short path for a rough pump and the servicability would be decades better.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all,
Does anyone have a dysfunctional HummerVI sputter coater, which could be cannibalized or purchased as a surplus item. I need vacuum pump (built in, type G-50D made by ULVAC-Sinku Kiko or equivalent) for this coater. Thank you in advance for your reply.
Chris Terlecki
Applied Analytical Sciences 3303 Harbor Blvd. Ste. H-4 Costa Mesa, CA 92626
id sma016866; Mon Apr 6 17:04:15 1998 Received: from elba.wadsworth.org (elba [172.16.1.100]) by newton.wadsworth.org (8.8.6.Beta3/8.7.1) with ESMTP id QAA11665; Mon, 6 Apr 1998 16:58:17 -0400 (EDT)
Dear Isabel, } } We're trying to make a deposition of 2 micron gold particles on a silica } substrate. } We cannot use an evaporator because we don't want a thin film: we want to } deposit the particles directly on the substrate, keeping them isolated and } without forming aggregates. } At first we dispersed the powder directly on the silica, but SEM analysis } showed they formed aggregates. } Then we've tried making a suspension on etanol and then putting a droplet } of the solution directly on the substrate. But still it formed aggregates. } } Does anyone know of a simple method to make such a deposit (without forming } aggregates) ? } If the particles were all of like charge, they might not aggregate. I suggest first making the substrate hydrophobic, if possible, then use an atomizer to spray a suspension of the particles in water onto the substrate. You will want to charge the droplets as they are leaving the atomizer (un- less simply using the atomizer works). I'm sure there's a way to charge the droplets--that's what Milliken (sp?) did with oil droplets to measure the electron's charge. Another possibility would be to deposit a thin film of wax on the substrate (with a conducting material also mixed in the wax) and propel the particles so that they stick to the wax. Good luck. Yours, Bill Tivol
An important first question is: What sample preparation method are you using to prepare your sapphire samples? To measure the thickness of the sapphire it is often useful to mount sacrificial silicon pieces around yo= ur otherwise transparent sample. Transmitted light is then used to monitor the color of the silicon which can be correlated to a thickness value. Th= is is done quite frequently when tripod polishing samples and is a very effective means of measuring the thickness. Of course, you will not be able to measure the thickness until you reach about 10 microns where the silicon starts to become translucent. This technique can also be used to=
ensure that you are polishing evenly across your sample.
We do have a mouse pad that I could send to you that provides a thickness=
scale for silicon as seen through both daylight and a tungsten filament i= f that would be helpful. =
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Brad Tinkham } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
I am preparing cross-sectional TEM samples of sapphire. I am trying to find a good way to measure the samples' thickness when I am under about 7= 0 micron. Any ideas?
Hummer VI coaters were orignally designed to accept several brands of vacuum pumps including the smallest Leybold, Welch and the Varian pumps. They can be bought easily on the used equipment market or from pump reconditioning companies. If you can't find one to fit internally, remote mount the pump and use a plastic vacuum hose to connect it to the baseplate. It doesn't hurt the system to have the pump remote mounted and you may find a larger pump on surplus easier than small one.
Good luck, Steve Collins South Bay Technology East
What is the purpose of your poly-L-lysine??? If it is simply to get sections to stick, then you would do well to switch to Fisherbrand Superfrost/Plus Microscopy Slides, catalog #12-550-15, FisherScientific, Pittsburgh, PA 15219. They are specially made for this purpose.
We use them exclusively and never lose sections, frozen or epoxy. They are more expensive than plain slides, but considering your time and your storage problems, you might like the convenience.
I have no commercial interest in this product--just a satisfied customer.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Orcein stains cartilage on paraffin sections a pretty distinct deep purple. You might check if it's neccesary to retain the cells, as they might be removed with alkali treatment (KOH?). I don't know what alkali does to orcein staining, but it would be easy to check. Best of Luck, Charlie Ginsburg NCC
} "I am currently looking for an optimal staining or sectioning procedure for } samples of ear and articular cartilage, whereby the stain is confined to } the extracellular matrix without staining (or very light staining) the } cells and nuclei within the cartilaginous lacunae. The reason for this is } to be able to accurately assess the ratio of extracellular matrix to } lacunar space by computerized morphometric analysis. Currently, } unavoidable staining of cells is obscuring the measurements made by the morphometric program, which uses gray scales and contrast to arrive at values". } } Further details I do not have. Any suggestions you may have on this matter } would be appreciated. } } } Neil Hand, } Nottingham, } England UK. } } ___________________________________________________________________ } Neil Hand } Department Histopathology, University Hospital, Nottingham NG7 2UH. } work : Tel: (0115) 924 9924 extension 43725 } _________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
A postdoctoral position is open at Northwestern University to work on In-situ deposition of Cubic Boron Nitride films, combining Ultra-High Vacuum Electron Microscopy with in-situ growth. Some details about the available hardware can be found at: http://www.numis.nwu.edu http://www.numis.nwu.edu/SINBAD and see also Collazo-Davila et al, Appl Phys Letts 72, 314-316 (1998) A strong background in thin film growth is required, and additional background in TEM and UHV will be highly relevant. Interested applicants should send a short CV via email to ldm3-at-apollo.numis.nwu.edu including email addresses for references.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 email: ldm3-at-apollo.numis.nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Paula Sicurello wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Listers, } } I am in need of a type 545 Polaroid camera back that has a frosted } glass where the film holder usually is. My SEM pix look OK on the screen } but if I make a print from the neg. it looks slightly out of focus. } I've been told that I need to focus the camera to match the screen } & this fancy camera back with the glass helps you do that. I know they } exist because I saw one on an ESEM. Does anyone know where I can buy one? } I called Polaroid and they were confused. } Can anyone help me with this problem? } } My future looks fuzzy & so do my prints, } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu Paula,
You didn't say what kind of SEM. The Japanese ones all use Mimya cameras. Others may be Graphlex compatible. You might try Calumet Camera in the Chicago area. I don't have the address, but they specialize in 4x5 and 8x10 cameras and have the frosted glass/freznel lens combination and hoods, or shades to make viewing easier.
You want to be sure of the compatability because the frosted glass MUST sit at the plane of the film.
Ken Converse Quality Images 16 Creek Rd. Delta, PA 17314 third party SEM service
Has anyone out there done any xNGF staining en grid, and could you possibly give me some guidelines for fixation and embedding, i.e., can you osmicate and what resins might be advisable. Many thanks Grace
I am the senior anatomopathologist of Paediatric Institute of Research of the University of Trieste(Italy). Due to two international collaborative research projects we ought to send to UK and USA(Baltimora) our frozen section of intestinal biopsies. These sections will be analyzed for EmA(antibidies antiendomysium) in Celiac Disease with Immunofluorescenze indirect and for cytokines TNF-alfa,INF-gamma in IBD with ISH. I fix the sections with acetone-cholrophormio(1:1) and store at -20=B0C (they last 2-3 months). I would like to know if there i= s any other technique to fix sections in a way I could store them at 2-6=B0= C and preserve them before searching for antigen for a longer period(such as 5-6 months).Yours sincerly Kind regards dr. Angelo Citt=E0 Dept.Anatomia Patologica Istituto per l'Infanzia 34100 TRIESTE e mail: acitt-at-tin.it
SUMMER I 1998 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-Section B)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A five week, Summer Session I 1998 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered four days per week (Monday through Thursday) between the hours of 8:00 am and NOON. Classes will begin on May 26 and end on June 25, 1998.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 per credit (for Nassau County residents or New York State residents with a certificate of residency).
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students.
If you have further questions, you should e-mail me directly at the address below.
For information about mail or telephone registration (Dial-a-Course) point your browser to http://www.sunynassau.edu/courses/sum98/sum98.htm. Telephone registration is only available until April 30, 1998.
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Like Woody White, I replaced the original pump but with a larger model and placed this under the bench; much easier to service and less backstreaming of vapour. The first Hummer with a triode sputter head suffered from sooting-up of the sputter head. At the time I was able to demonstrate to the manufacturer that the sooting-up problem was caused by the mineral oil and could be avoided by using Fomblin rotation pump fluid. It's ancient history now but the problem seemed severe at the time and when I was replacing the pump for the third time I was consoled that "we have to replace over a hundred pumps." The original correspondent needs to replace the original pump, and well he might. However, some people may only need to have the right pump fluid and a decent size pump. The original pump's capacity was too low. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
_____________________________________________________________} } Does anyone have a dysfunctional HummerVI sputter coater, which could be } cannibalized or purchased as a surplus item. I need vacuum pump (built } in, type G-50D made by ULVAC-Sinku Kiko or equivalent) for this coater. } Thank you in advance for your reply. } } Chris Terlecki } } Applied Analytical Sciences } 3303 Harbor Blvd. Ste. H-4 } Costa Mesa, CA 92626 } } ph: 714-434-6894 } fax: 714-434-0294 } email: aas-at-pacbell.net }
Re: sputter coater pump _____________________________________________________________} } Does anyone have a dysfunctional HummerVI sputter coater, which could be } cannibalized or purchased as a surplus item. I need vacuum pump (built } in, type G-50D made by ULVAC-Sinku Kiko or equivalent) for this coater. } Thank you in advance for your reply. } } Chris Terlecki } } Excuse my ignorance re: details of pumping system for sputter coaters but some respondent have commented on backstreaming of pump oil. We use mechanical pumps on visible light microscope cathodoluminescence attachments using a cold cathode discharge and always use a simple foreline trap (copper gauze type - ca$300.00) to reduce the backstreaming. Yearly maintenance for trap is less than $50.00. Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
I wish to receive notice about the book "Manual of microscopic Analysis of Feedstuffs. 3rd Ed. The Amer. Assoc. of Feed Microscopists, 1922, p. 73-93" or the articles in the same book, autors BATES L.e coll. Feed ingredient descriptions of animal origin.
A customer in the Chicago, IL area has asked for help locating service for their HNU EDS system. They are having trouble getting service from the parent company, and would like to locate a third party service organization. Any information would be greatly appreciated.
Regards, Chris Wadelton Amray, Inc. (800) 591-8791
Advanced Surface Microscopy would like to buy used Digital Instruments NanoScope equipment of the following types: NanoScope II with AFM (contact mode) NanoScope III with contact and/or Tapping Mode AFM and other related equipment.
Please contact me offline, since I am not a regular subscriber to this listserver.
Thanks very much. Don Chernoff
Advanced Surface Microscopy, Inc. E-Mail: asm-at-indy.net 6009 KNYGHTON RD. Voice: 317-251-1364 INDIANAPOLIS IN 46220 Toll free: 800-374-8557 (in USA) web: http://www.a1.com/asm Fax: 317-254-8690 (note: "a1"= letter "a", numeral "1")
The AMRAYINC-at-aol.com address was removed from the listserver this morning because Amray is installing a new E-mail exchange system. The addresses will be changing, in the interum Amray is still available via E-mail at the above address as well as AMRAYSERV-at-aol.com (service) and AMRAYSALES-at-aol.com (sales).
Dear list, Similar question was discussed not so far, but I saw no reliable solution. I'm sorry if I simply missed it. The problem is to know diffraction orientation relative to the object feature. I can defocus diffraction spots and see images of the feature (edge for example) in each spot but I can defocus two sides and get opposite directions. Which one is true? Or there is another method? I do not need to calibrate rotation angle as it's always 0 (or 180deg?) at JEM2010. I want to know definitely if it 0 or 180. We need this to determine SiC and AlN polarity.
Hello to all, We have been using another department's double glass distilled water, and their still is down. Our building has a house "distilled" water tap. I called to find out the method by which this water is purified, and was told that the water is softened, passed through reverse osmosis, deionized, filtered, UV sterilized, and delivered to the building at about 15 megaohms of resistivity. This leads me to my question, what are the standards by which other labs (biological EM) gauge their water? Distilled vs deionized? Your opinions or a reference citing would be greatly appreciated.
-- Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
In the recent emails regarding frosted glass, noone has mentioned why these work well. I have been unable to find out why from several reference works, so does anyone know? Is it that the uneven surface exaggerates the out-of-focus appearance since there is then no even plane of focus on the glass?
Doug Darnowski
****************************************************************************** Douglas Darnowski Department of Crop Sciences 384 ERML 1201 West Gregory Drive University of Illinois Urbana IL 61801 work ph: (217) 244-6150 fx: (217) 333-4777 home ph: (217) 356-6606 fx: (217) 356-4454 email: darnowsk-at-staff.uiuc.edu
I think that you can use the shadow image in a convergent beam pattern. Underfocus the crossover and your shadow image in the BF disk lines up with the diffraction pattern.
You can check this with GaAs from a known wafer because the orientations are determined by etching the sample. The data sheet that comes with the wafer should let you know what the absolute directions are using the major and minor flats. You can prepare your sample very easily and maintain the crystallographic orientation of the sample with the small angle cleavage technique. John McCaffrey and I talk a little bit about this in our paper on SACT in the MRS TEM Sample Prep IV (Vol 480).
I may be mistaken, but I also thought that the 180 degree ambiguity was taken care of in the 2010.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Dear list, Similar question was discussed not so far, but I saw no reliable solution. I'm sorry if I simply missed it. The problem is to know diffraction orientation relative to the object feature. I can defocus diffraction spots and see images of the feature (edge for example) in each spot but I can defocus two sides and get opposite directions. Which one is true? Or there is another method? I do not need to calibrate rotation angle as it's always 0 (or 180deg?) at JEM2010. I want to know definitely if it 0 or 180. We need this to determine SiC and AlN polarity.
The frosting merely serves to scatter light from the film plane rather so that you can see the image. If it were not frosted, the light would pass thru - the glass would be invisible, and you would see no image.
If you could look at the image from the lens side you could focus the camera using an opaque material in place of the film. But since cameras are sealed up, they use the frosted glass so you can see an image from the back side.
At 11:30 AM 4/6/98 -0600, you wrote: } } In the recent emails regarding frosted glass, noone has mentioned why these } work well. I have been unable to find out why from several reference works, } so does anyone know? Is it that the uneven surface exaggerates the } out-of-focus appearance since there is then no even plane of focus on the } glass? } } Doug Darnowski
A student in my laboratory has a number of digital images of gels and transmission electron micrographs which she would like to include in her thesis. I know that prints from our inkjet printer have poor archival properties (they turn brown over a period of a year or so, depending on light, air exposure), but we do have access to a dye sublimation printers elsewhere. How long can we expect black and white dye sublimation prints to last in a thesis without discoloring or fading? Given that these printers haven't been around very long, does anyone actually know?
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
At 03:16 PM 4/7/98 -0400, MARIE CANTINO wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I posed this question to a Kodak rep at the last MSA in Cleveland. He stated the prints were archival; that is, they should last at least as long as properly fixed photographic material, at least 50 years.
Rick A. Harris, Director Microscopy and Image Analysis Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 752 3085 fax raharris-at-ucdavis.edu
} -----Original Message----- } From: MARIE CANTINO [mailto:CANTINO-at-ORACLE.PNB.UCONN.EDU] } Sent: Tuesday, April 07, 1998 12:17 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Archival properties of dye sublimation prints } } } -------------------------------------------------------------- } .... } } A student in my laboratory has a number of digital images of gels and } transmission electron micrographs which she would like to include in her } thesis. ... we do have access to a dye sublimation printers } elsewhere. How long can we expect black and white dye } sublimation prints } to last in a thesis without discoloring or fading? ...
My experience with dye-subs is that grayscale prints are usually a result of CMY printing, although two possibilities are (1) the CMY ribbon also had a "K" of black component, or (2) a monochrome "K" ribbon was used. If the prints were a result of "K" printing then they are not likely to change color, but they may fade while the paper stock remains white. If they are a result of CMY printing, they are liable to acquire a pink or green tint (... not bad though ...).
Lastly, and at least for Kodak dye-sub printer engines, there exists a ribbon/paper combination which lamenates and protects the image from UV ... or you can try to find UV protection, usually as a form of a spray, to apply to print.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
It probably depends on what dye sub printer you have as to how well a print will stand up to the passage of time.
A Kodak printer we had put a plastic coat on top of the dye as the last step in the printing process.
An old Sony dye sub printer I had did not do this. The dyes would smear if any moisture at all got on them. Also after several months in the hallway under fluorescent light, the dyes would fade. If they were not exposed to light, they kept their color 4-5 years and I suspect they would keep much longer.
I do not know whether the plastic coating prevents the fading of the dye when exposed to light a number of months because I haven't tested any of these prints.
That is the extent of my knowledge on the subject. Hope it helps.
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 73-384 CHS 951761 Los Angeles, CA 90095-1761
} ---------- } From: } CANTINO-at-ORACLE.PNB.UCONN.EDU[SMTP:CANTINO-at-ORACLE.PNB.UCONN.EDU] } Sent: Tuesday, April 07, 1998 12:16 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Archival properties of dye sublimation prints } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } A student in my laboratory has a number of digital images of gels and } transmission electron micrographs which she would like to include in } her } thesis. I know that prints from our inkjet printer have poor archival } properties (they turn brown over a period of a year or so, depending } on } light, air exposure), but we do have access to a dye sublimation } printers } elsewhere. How long can we expect black and white dye sublimation } prints } to last in a thesis without discoloring or fading? Given that these } printers haven't been around very long, does anyone actually know? } } Marie } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology, U-131 } University of Connecticut } Storrs, CT 06269 } Ph: 860-486-3588 } Fax: 860-486-1936 } } }
I would be happy to help. I was approached by HNU several years ago to sell their instruments, and after visiting their manufacturing facilities and meeting with their top brass, I decided that they were lacking in many respects and I didn't choose to go with them. I would be happy to help anyone, as best I can, who chose this manufacturer as their intrusion into the market was ill-planned.
They assumed that the US manufactured EDS detector combined with the German produced PC-based software would give them some market advantage, and they were right for a very short time. But the manufacturers who were more heavily invested in the art of EDS soon translated their DEC LSI-11 based software into PC based applications, and HNU was left out to dry as those manufacturers produced PC based applications that worked well and offered the market appeal of known market brands in the industry.
} Dear All, } } A customer in the Chicago, IL area has asked for help locating } service for their HNU EDS system. They are having trouble getting } service from the parent company, and would like to locate a third } party service organization. Any information would be greatly } appreciated. } } Regards, } Chris Wadelton } Amray, Inc. } (800) 591-8791 } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
I wish to receive notice about the book "Manual of microscopic Analysis of Feedstuffs. 3rd Ed. The Amer. Assoc. of Feed Microscopists, 1992, p. 73-93" or the articles in the same book, autors BATES L.e coll. Feed ingredient descriptions of animal origin.
I wish to receive notice about the book "Manual of microscopic Analysis of Feedstuffs. 3rd Ed. The Amer. Assoc. of Feed Microscopists, 1992, p. 73-93" or the articles in the same book, autors BATES L.e coll. Feed ingredient descriptions of animal origin.
Marie, Archival properties depend on your particular printer and the printing ribbon used.
We are on our 2nd generation dye-sublimation printer. Our first was purchased ~ 6 years ago when the technology was very new. Greyscale prints from this printer were made from an overlay of RGB color and tended to have a purplish cast to them. The colors did fade when exposed to UV light but were fine if kept in the dark the majority of the time. At that time there was no product with the transparent UV-protectant overlay available.
We recently upgraded with a Codonics 1660 dye-sub printer. Two of the main features I looked for before purchase was that there was a black and white ribbon available and a color ribbon that incorporated the UV protection overlay. This printer has both. The use of the black and white ribbon eliminates the chance of any misalignment of the color sheets to get true greyscale without hints of another color. Also available are RGB ribbons with (approx. $3/8x10" sheet) or without (approx. $2/sheet) the UV-protection laminate. We often use the less expensive color ribbon when output is not intended for long term use and then switch to the ribbon with overlay for final copies.
We have been assured that prints produced with the black and white or color + overlay ribbon are indeed archival but time will tell!!
Debby Sherman, Manager Microscopy Center in Agriculture Purdue University --------------------------------------
A student in my laboratory has a number of digital images of gels and transmission electron micrographs which she would like to include in her thesis. I know that prints from our inkjet printer have poor archival properties (they turn brown over a period of a year or so, depending on light, air exposure), but we do have access to a dye sublimation printers elsewhere. How long can we expect black and white dye sublimation prints to last in a thesis without discoloring or fading? Given that these printers haven't been around very long, does anyone actually know?
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
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Fellow Scanners,
I have a Sharp 2400 DPI Flatbed which I am thrilled with but Sharp has gotten out of the scanner market. So, whats the deal? Our ceramics department needs to purchase a flatbed for TEM negatives, slides, positives color B&W etc....Which one? The esteemed Dr. Mark Farmer from the University of Georgia has suggested a Umax; I know and respect Dr. Farmer and usually follow his recomendations so I would love to find the North East distributer for Umax or if anyone has another flatbed choice for our ceramics department I would appreciate a reply.
48 degrees, dreary, and it smells funny in several areas... just thought you'd like to know! John Grazul Rutgers University Electron Imaging Facility
High resolution (nanometer) imaging Protein folding force measures Protein binding force measures
This technique is still new enough that it requires a skilled microscopist, but is certainly breaking new ground in microscopy.
At 11:13 AM 3/27/98 -0700, Gillian Bond wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
you mention smudging with ink-jet inks. I have started using an Epson Stylus 600 and I notice that it doesn't smudge if you use the full glossy papers. I don't know about fading, though but it may be useful to use the glossy finish for more permanent results if your printer can use it and the cheaper papers for routine work.
Malcolm Haswell University of Sunderland UK ----------
Marie,
It probably depends on what dye sub printer you have as to how well a print will stand up to the passage of time.
A Kodak printer we had put a plastic coat on top of the dye as the last step in the printing process.
An old Sony dye sub printer I had did not do this. The dyes would smear if any moisture at all got on them. Also after several months in the hallway under fluorescent light, the dyes would fade. If they were not exposed to light, they kept their color 4-5 years and I suspect they would keep much longer.
I do not know whether the plastic coating prevents the fading of the dye when exposed to light a number of months because I haven't tested any of these prints.
That is the extent of my knowledge on the subject. Hope it helps.
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 73-384 CHS 951761 Los Angeles, CA 90095-1761
} ---------- } From: } CANTINO-at-ORACLE.PNB.UCONN.EDU[SMTP:CANTINO-at-ORACLE.PNB.UCONN.EDU] } Sent: Tuesday, April 07, 1998 12:16 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Archival properties of dye sublimation prints } } A student in my laboratory has a number of digital images of gels and } transmission electron micrographs which she would like to include inb her } thesis. I know that prints from our inkjet printer have poor archival } properties (they turn brown over a period of a year or so, depending on } light, air exposure), but we do have access to a dye sublimation printers } elsewhere. How long can we expect black and white dye sublimation prints } to last in a thesis without discoloring or fading? Given that these } printers haven't been around very long, does anyone actually know? } } Marie } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology, U-131 } University of Connecticut } Storrs, CT 06269 } Ph: 860-486-3588 } Fax: 860-486-1936
by mail450.icon.co.za (8.8.8/8.8.8) with SMTP id RAA14537; Wed, 8 Apr 1998 17:03:02 +0200 (GMT) Received: by localhost with Microsoft MAPI; Wed, 8 Apr 1998 17:05:26 +0200 Message-ID: {01BD6310.86D0CF60.anaspec-at-icon.co.za} "'MSSA listserver'" {MSSA-at-UCTBC1.UCT.AC.ZA}
Hi all.
We were wondering if anybody out there may have a spare XP3 pulse processor we could by at a good price ? We have a specific need for high count rates off an Oxford detector. The XP3 has a fast process time of 2.5?sec which is ideal.
Thanks.
Luc Harmsen Anaspec, South Africa Technical support for E.M. operators, world wide. anaspec-at-icon.co.za TEL: ++ 27 (0) 11 476 3455 FAX: ++ 27 (0) 11 476 7290
Meeting Announcement San Francisco Microscopical Society
Thursday, April 9, 1998 Rockridge Branch, Oakland Public Library 7:00 P.M.
"Microscopic Pond Life" by Rick Ellis
Rick Ellis is a microscopist and photomicrographer whose talks we have all enjoyed in the past. This promises to be another fascinating evening as Rick takes us on an exploration of the rich and varied world of pond microorganisms. This is the type of microscopy that is readily available to everyone (especially in this year of El Ni=F1o). We will have a Saturday Workshop later this year to collect and examine pond critters, so this will be a good opportunity to see what you can expect to find, and how to prepare the samples for examination. Please join us!
Further Particulars:
http://www.microdataware.com/sfms
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
We recently purchased a Umax Powerlook III (a new model, about $2k U.S.). They call it a "42-bit" scanner through a bit enhancement technology, and it has a real resolution of 1200 x 2400 dpi. At any rate, it seems to work pretty good for both negatives and prints. The software is not overly intuitive and we received no book on how to use it but we have generally worked out the kinks and are fairly happy. It won't capture everything on your negatives since their grain size is smaller than the available resolution and their dynamic range is higher than what we can get (3.4, I think), but it works well for those times when you don't want to print or need to bring out a feature on the negative.
Their web address is : http://www.umax.com/ and it contains links to resellers in specific regions of the U.S. (and perhaps internationally; I didn't look).
Hope this helps.
Cheers,
John Vetrano
----------
Fellow Scanners,
I have a Sharp 2400 DPI Flatbed which I am thrilled with but Sharp has gotten out of the scanner market. So, whats the deal? Our ceramics department needs to purchase a flatbed for TEM negatives, slides, positives color B&W etc....Which one? The esteemed Dr. Mark Farmer from the University of Georgia has suggested a Umax; I know and respect Dr. Farmer and usually follow his recomendations so I would love to find the North East distributer for Umax or if anyone has another flatbed choice for our ceramics department I would appreciate a reply.
48 degrees, dreary, and it smells funny in several areas... just thought you'd like to know! John Grazul Rutgers University Electron Imaging Facility
I have used both Umax and Microtek brand scanners (SCSI I/O). No problems with either. They can be purchased from any number of computer suppliers like Computability, CDW, Microsystems Warehouse, B&H Camera, etc.
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Fellow Scanners,
I have a Sharp 2400 DPI Flatbed which I am thrilled with but Sharp has gotten out of the scanner market. So, whats the deal? Our ceramics department needs to purchase a flatbed for TEM negatives, slides, positives color B&W etc....Which one? The esteemed Dr. Mark Farmer from the University of Georgia has suggested a Umax; I know and respect Dr. Farmer and usually follow his recomendations so I would love to find the North East distributer for Umax or if anyone has another flatbed choice for our ceramics department I would appreciate a reply.
48 degrees, dreary, and it smells funny in several areas... just thought you'd like to know! John Grazul Rutgers University Electron Imaging Facility
Debby Sherman wrote: ========================================== Unicryl is made by BBInternational and distributed in the USA by Vector Laboratories, Inc. (1-800-227-6666). They are a CA based company and are not open yet today. I did call there yesterday and asked for a method booklet on Unicryl and the individual who I talked to did not indicate that the resin was unavailable. =========================================== The embedding resin, Unicryl(TM), is the product of the following:
British Biocell International Ltd. Golden Gate Ty Glas Avenue Cardiff CF4 5DX UK
SPI Supplies has imported Unicryl resin into the USA almost since its inception as a product and it is now also available from a number of the other suppliers of chemicals to microscopy laboratories, for example, Pella and EMS.
The debate on which embedding resin is or is not "better" will probably still be going on long after we are all gone from this word. At least some of our customers have reported excellent results with Unicryl but the one thing that does seem to stand out is that there is a perception among those who are sensitized (from a dermatitis standpoint) to this class of acrylic resins is that the Unicryl system is "milder" than the others. Now I will be the first to admit that this is not a very scientific sampling but if one should be listening to their customers, this is one thing at least some of them have been telling us.
I for one would welcome any kind of dialogue describing the experiences people have had with these different resin systems. If some are "better" or "worse" from a dermatological standpoint, it would be good to have the benefit of the collective experience of those working with these different media.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Hypothetically, if I had a fully loaded, 1 year old Kevex Sigma 4 system available (no detector), would anyone be interested in purchasing it and what would be a fair asking price. The system was purchased for $75,000 1 year ago.
Wayne Kaboord Materials Research Specialist Eaton Corporation Innovation Center 4201 North 27th Street Milwaukee, WI 53216 414-449-7783 WayneKaboord-at-eaton.com
id xma023997; Thu, 9 Apr 98 08:17:11 -0400 Received: from charlotte.cpt.afip.org (charlotte.cpt.afip.org [10.20.30.46]) by elon.cpt.afip.org (950413.SGI.8.6.12/950213.SGI.AUTOCF) via ESMTP id IAA16246; Thu, 9 Apr 1998 08:22:55 -0400 Received: from localhost (oliver-at-localhost) by charlotte.cpt.afip.org (950413.SGI.8.6.12/950213.SGI.AUTOCF) via SMTP id IAA05283; Thu, 9 Apr 1998 08:22:15 -0400
Please note that this is a personal observation and does not reflect the opinion of my office nor of any group associated with me.
I use a Umax Powerlook II as the workhorse machine in my lab, and have been generally happy. We will likely be moving to one of the newer machines soon. However, you should look at some reviews in the consumer press before making a final decision. Some folk in the lab next to me use an Agfa scanner, and they are happy with it.
The biggest criticism of the Umax scanners that I have seen in the press is that they sometimes do not give as great a depth or discrimination of tone as some competitors. For instance, if you place a sheet with 256 greyscale values in the scanner, it may only give you 200 values. I tried something like that with my scanner after I read the review and noticed the following:
If I took 256 values from black to white I got about 240 values out, which was pretty good. If, however, I took 256 values from dark grey to light grey, it did much more poorly -- about 150 values. In other words, there was no way to calibrate the illumination or sensitivity of the sensor to adjust for a dingy image.
In one sense, the scanner was "correct" in that I put in a dingy image and got dingy results. However, if one wants to take data acquired from a scanner and do image processing, one wants as much data as possible from that scanner. And that means that one would like to fill those 256 levels with whatever is available.
Ideally, one would want to be able to modify the illumination of the scanner to get a full 256 tone values out of an image, regardless of what the brightest and darkest regions are. The "histogram stretching" functions of the TWAIN driver software do *not* accomplish this; instead it simply does a stretch *after* data acquisition (at least as I have observed). Thus, you still get only, 150 grey values instead of 256, but they are stretched between 0 and 255.
This may not be a problem for you, but since I do image processing, I am very interested in getting as many tone levels as I can out of an image. I am *not* sure if competitors are all that much better, but you might want to look at some of the reviews which actually do quantitative measurements before you buy.
billo
On Wed, 8 Apr 1998, John Grazul wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Fellow Scanners, } } I have a Sharp 2400 DPI Flatbed which I am thrilled with but Sharp } has gotten out of the scanner market. So, whats the deal? Our } ceramics department needs to purchase a flatbed for TEM negatives, } slides, positives color B&W etc....Which one? The esteemed Dr. Mark } Farmer from the University of Georgia has suggested a Umax; I know } and respect Dr. Farmer and usually follow his recomendations so I } would love to find the North East distributer for Umax or if anyone } has another flatbed choice for our ceramics department I would } appreciate a reply. } } } 48 degrees, dreary, and it smells funny in several areas... } just thought you'd like to know! } John Grazul } Rutgers University } Electron Imaging Facility }
In response to Bill's comments, I have both an Agfa and two UMax scanners and I personally love the Umax's and would never buy another Agfa. Both give acceptable images and neither is consistently perfect. The difference is in the software. Trying to manipulate gain and black level on the Agfa is complicated and time consuming, whereas the Umax is fast and intuitive. My first UMax is still working 9 years later and still gives fine images. Dave
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Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
I am a faculty member at Brookdale Community College in Lincroft, NJ who operates a Zeiss 9C TEM which was donated to the College. Unfortunately, however, we do not have a knife maker or ultramicrotome for tissue preparation. Does anyone know of a used ultramicrotome and knife maker which can be purchased for a reasonable price? (or perhaps even donated to the College) Thanks in advance for your help.
Mike Palladino
Michael A. Palladino, Ph.D. Instructor of Biology Brookdale Community College Voice: (732) 224-2871 E-mail: mpalladino-at-brookdale.cc.nj.us
Currently I work as a corporate web designer and use Photoshop imaging software daily for the past 4 years. I am concerned as to how to combine my visual imaging skills with my interest in natural and physical science (so-so on medical). At 44, I am intimidated by the notion of schooling for many years. What is entailed and where can I ask about a microscopy program in New York City. I believe the visual aspect of scientific analysis is fascinating.
I am volunteering web pages for the invertebrate scientists at the American Museum of Natural History and have seen their electronic microscope equipment (I was asked to teach Photoshop so that the scientists could color the scans) and other imaging equipment and 3-d software. Thrilling!! I treasure my book on microscopic art (Felice Frankel). Also, I perform nature photography when away from the city.
Thank you in advance for any education or vocation advice you have time to give.
Renee Recker 16 W. 16th St. NYC, NY 10011 212-675-1665
some samples of web pages for Amer. Museum of Nat. History: http://www.renorex.com/invertebrateshome http://research.amnh.org/~mikkel
We have two Leitz metallographs and are having problems with the 450 watt xenon lamphouses on both.
One has operated fine for the last 15 years (same bulb!) Recently, I noticed an odor after about 15 minutes of continuous use, perhaps a burning phenolic. We replaced the transformer in the high voltage unit, but the odor persisted. Predictably, something finally blew and now the bulb will not ignite.
The other metallograph was fine and then one day the xenon bulb would not ignite. We replaced the bulb and it still fails to ignite (though the high voltage unit attempts ignition).
Are there any optical service engineers in the Chicago area knowledgable in xenon units. The bulbs are high pressure and hazardous to the uniformed. I suspect that there are either connection problems or blown components in the high voltage units.
If anyone can help, then please reply off-line or call me directly.
Thank you.
Alan Stone ASTON Metallurgical Services 773/528-9830
We have recently made some changes to our web site. Please visit us at www.rjlg.com. We would greatly appreciate any comments or opinions you may have pertaining to your visit.
Keith Rickabaugh Manager, Materials and Particle Characterization {krickabaugh-at-rjlg.com}
RJ Lee Group, Inc. 350 Hochberg Road Pittsburgh, PA 15146 ph: 724-325-1776 www.rjlg.com
Hey there imaging buffs 1) Is there a way to create the red/green 3D images using stereo pair TEM mages without buying an expensive 3D imaging program? 2) What thickness of sections would be needed for TEM work to get the effect of 3D depth? 3) What is the best angle? I would be using a Philips 410LS w/goniometer tilt stage. I'm going to hit the library but tips from the horses mouth are always much better. Thanks in advance.
Isabel Nogueira wrote: ================================================== We're trying to make a deposition of 2 micron gold particles on a silica substrate. We cannot use an evaporator because we don't want a thin film: we want to deposit the particles directly on the substrate, keeping them isolated and without forming aggregates. At first we dispersed the powder directly on the silica, but SEM analysis showed they formed aggregates. Then we've tried making a suspension on etanol and then putting a droplet of the solution directly on the substrate. But still it formed aggregates.
Does anyone know of a simple method to make such a deposit (without forming aggregates) ? =================================================== There are three ways I could suggest, not based on experience specifically with gold particles, but by extrapolation from the dispersal of other fine particles and powders:
a) Camphor/naphthalene dispersion method (I described this previously and it should be available in Nestor's archives), where by the powder to dispersed is added to a 60% camphor/40% naphthalene (the eutectic composition of the system) which becomes a liquid at a few degrees above room temperature. Putting a drop of the suspension on your substrate will result in its instantly freezing. Put substrate and all into a vacuum over night (mechanical pump is enough) and by morning, the camphor/naphthalene would have sublimed away leaving the gold particles nicely and finely dispersed. You would regulate the degree of coverage by controlling the loading of the eutectic solution and the amount deposited.
b) Dispersion in an organic solvent but add a small amount of Parlodion (R) or perhaps any of the other similar in composition materials (e.g. Collodion (R)). After dispersal on the substrate, you would remove the organics with a barrel-geometry plasma etcher. The gold particles should be left in situ and not agglomerate. The adhesion to the substrate with (a) might be better than with (b).
c) Variation on the above, but using water, which is then applied to the substrate while the substrate is below the water freezing point. The ice is then sublimed away over night in a freeze fracturing device, leaving the gold particles dispersed without agglomeration.
However, some powders just do not want to disperse, perhaps because they are to some degree aggregates and not simply agglomerates. Being able to use a method that can be tightly controlled, such as (a) or (b), is sometimes the only way to discriminate between different samples where their only real differences happen to be their degrees of aggregation (vs. agglomeration). Over the years we have seen on occasion, large (I call 2 um "large") gold particles as in thick film paste systems that have been doublets and triplets, etc. so it is not impossible that at least some of your problem could be that not all of the particles are in fact single individual particles.
Disclaimer: Structure Probe, Inc. performs these kinds of services for clients as a regular part of our business.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Could a local air conditioning/refrigeration/plumbing company help? How about an appliance repair shop? Brazing companies? Heat exchanger builders/rebuilders? Automobile radiator repair shop? Local technical university?
My $0.02
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
Conceptually it should be quite possible and straightforward.
We collect the left and right B/W images using a 4 to 6 degree tilt between the views for our SEM work. Much more than 6 degrees gets unnatural. It represents crossing the eyes too much.
Since I work with the SEM, I don't know what the sample requirements would be for TEM. I would think as long as you have some topographic relief you should be okay. Even layers of atoms might be enough if you are at high enough mag.
Once you have the images, you can easily change the hues for the one from gray to red and the other from gray to green or blue using programs like PhotoShop. Preparing the anaglyph simply requires merging the two colored images into one true-color image. Since one image would supply the red intensity and the other the green intensity, it should be straightforward - I can think of how I would write a program to do it. But maybe the Photoshop gurus can tell what buttons to push to make it happen.
At 06:02 PM 4/9/98 -0500, you wrote: } Hey there imaging buffs } 1) Is there a way to create the red/green 3D images using stereo pair } TEM mages without buying an expensive 3D imaging program? } 2) What thickness of sections would be needed for TEM work to get } the effect of 3D depth? } 3) What is the best angle? I would be using a Philips 410LS } w/goniometer tilt stage. I'm going to hit the library but tips from the horses } mouth are always much better. Thanks in advance. } } Rick Vaughn } RLVAUGHN-at-MAIL.UNMC.EDU ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
I have had more success with SEM stereo pairs! The best results from TEM came from 0.1-0.5 um-thick sections. All tilted about 6 degrees. I have not done a lot of TEM because it was never very effective with normal ultrathin sections. It may be better with freeze fracture replicas and other specimens on formvar/carbon films such as direct prep. cells etc.
Take the stereo pair of images. Make good sized prints. Then get a friendly photgraphic person to photograph one print using a red filter onto colour slide film. Then double expose with the other print - in other words, photograph the second print onto the same film using a green filter. You then have effective stereo which you can show at meetings using red/green glasses and only one projector.
There is a convention which I forget regarding which way around you do this so that you hold the glasses properly. I'm replying to this from home, also my friendly photographer is skiing in Canada at the moment! Write back if you need the details (and if he survives Lake Louise or wherever he's gone this time!).
Keith Ryan Plymouth Marine Lab., UK
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Hey there imaging buffs 1) Is there a way to create the red/green 3D images using stereo pair TEM mages without buying an expensive 3D imaging program? 2) What thickness of sections would be needed for TEM work to get the effect of 3D depth? 3) What is the best angle? I would be using a Philips 410LS w/goniometer tilt stage. I'm going to hit the library but tips from the horses mouth are always much better. Thanks in advance.
I use Photoshop to do this in my bio TEM class, but you should be able to do it in NIH Image (can't beat the price). You could theoretically do on any section thickness, the effect depending on the magnification of the structure viewed. You make each image a different color, adjust the transparency and overlay them. You can view them on screen or output them to various hard copy media.
I usually have the students use their replicas (usually freeze-fractured yeast) since these have a 3-d topography, but I've also done this with chloroplasts in "thick" (about 250nm) section (don't stain them if you've used OsO4); they are just O.K. at 100kV. Depending on what you want to see, you can leach out a lot of background density by using KMnO4 and maybe work with thicker sections. More volts should also help as would an energy filter.
The best angle is dependent on the effect desired. To get an idea of the range of sizes and angles you can work with you might get a hold of Heuser, 1989 Journal of Electron Microscopy Technique 13: 244-263 and Steere, R.L. In Chapter 5 of Current Trends in Morphological Techniques Vol 2 CRC Press 1981. These deal mostly with conventional stereo pairs and replicas but the approach and geometry is similar .
cheers, John Heckman TEM Supervisor Center for Electron Optics Michigan State University} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hey there imaging buffs } 1) Is there a way to create the red/green 3D images using stereo pair } TEM mages without buying an expensive 3D imaging program? } 2) What thickness of sections would be needed for TEM work to get } the effect of 3D depth? } 3) What is the best angle? I would be using a Philips 410LS } w/goniometer tilt stage. I'm going to hit the library but tips from the horse s } mouth are always much better. Thanks in advance. } } Rick Vaughn } RLVAUGHN-at-MAIL.UNMC.EDU }
For the first question, here is a good starting point on the Web: http://www.tisco.com/3d-web/
There is a link called "how to make..." but it wouldn't come up. I don't know if that page is gone or just down temporarily. Basically you probably need to use Adobe Photoshop. We have used that quite successfully in making these images. There may be other ways, but if you want to do it digitally, you need a program that can separate the color channels.
My colleague with the good notes on the second two questions is out of town but if I don't see other answers forthcoming I'll have him contact you when he gets back. He images dislocation stereo pairs often.
Cheers, John Vetrano john.vetrano-at-pnl.gov
----------
Hey there imaging buffs 1) Is there a way to create the red/green 3D images using stereo pair TEM mages without buying an expensive 3D imaging program? 2) What thickness of sections would be needed for TEM work to get the effect of 3D depth? 3) What is the best angle? I would be using a Philips 410LS w/goniometer tilt stage. I'm going to hit the library but tips from the horses mouth are always much better. Thanks in advance.
I use an Agpha-Arcus ll and I'm very satisfied with the results it gives me, since we need grey scale for triblock copolymers. There is a newer vertion of the scanner which might have better features. Regards,
The plans for the CSMS/MIKMAS meeting are coming along. We will have everything from digital imaging workshops to a demo of Virtual TEM usage. There will be venders, and several of them will have live demo's of their equipment.
I hope to have the schedule out around next weekend.
For those of you who plan to attend, the directions to our facility are now on the web and can be found below:
I will also post the reults of the show and supper survey next week.
Please consider coming. We are going to set up a block of rooms at University prices at the Raddison Hotel just off of Neil Street in Champaign. Please give us until next Tuesday to complete the reservation.
Radisson Hotel #: 217-398-3400 The Hotel location is marked on the Web Page
If you have any Questions please call 217-244-1567 between 7:30am - 4pm.
**All those considering presenting, please contact me this week if you contacted me via mail , email or phone so far**
Lou Ann -- *************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
Microscopy Home Page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/CSMS/csms.html
Personal Home Page: http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/LAM.html
I currently am trying to find a way to image composite fibers of 50 to = 500 micron in length and 10 micron in width with out them crossing and = touching. =20
The idea is to image, threshold, and then count and measure these = fibers. right now we have to go through each field and manully measue = them. This can be quite tedious and time consuming when there are = thousands involved. We can use the image analyser to do fields = automatically with some manual interaction.
Can anyone suggest a means of dispersing such fibers in the preperation = process that will not allow them to touch and overlap??? We can also = set the image analyser to randomly eliminate the ones that touch. This = is an alternative but would like to be able to measure them all, if = possible. Also this is done on just basic brightfield and does not = require any special illumination such as polarized light.=20
Any suggestions can be sent to this address and I will reply. Thank = You.
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} Dear Fellow = Microscopist, {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} I am in need of someone = assistance. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} I currently am trying to find a way = to image=20 composite fibers of 50 to 500 micron in length and 10 micron in width = with out=20 them crossing and touching. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} The idea is to image, threshold, and then count and = measure=20 these fibers. right now {/FONT} {FONT size=3D2} we have to go = through each=20 field and manully measue them. This can be quite tedious and time=20 consuming when there are thousands involved. We can use the image = analyser=20 to do fields automatically with some manual interaction. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Can anyone suggest a means of dispersing such fibers = in the=20 preperation process that will not allow them to touch and = overlap??? We=20 can also set the image analyser to randomly eliminate the ones that = touch. =20 This is an alternative but would like to be able to measure them all, if =
possible. Also this is done on just basic brightfield and does not = require=20 any special illumination such as polarized light. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Any suggestions can be sent to this address and I = will=20 reply. Thank You. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Cono = Passione {/FONT} {/DIV} {/BODY} {/HTML}
Funny you should ask this question. For the last two semesters I have been working on just this sort of thing with the SEM. I had fairly good success using Photoshop. The trick is making sure that your images are alligned vertically, and playing around with the color until it matches the color of the filters on the viewerlenses. As far as the tilt angle, that depends on the magnification and the degree of roughness of the surface being imaged. Generally speaking, a smoother sample will require more tilt (7-15 degrees) than a rough one (3-7 degrees). At higher magnification, too much tilt can mean too much parallax (displacement) and the result may be uncomfortable to view. Usually 4-10 degrees of tilt will produce sufficient parallax. If still unsure, you can do what I did and take a series of images, each at 3 degree differences in tilt. Just be sure that when you refocus the image after tilting that you do so without changing the magnification. On the SEM this means using the Z axis control instead of the objective lense control. Also be careful when you tilt that you maintain your field of view (tracing the outline of some major features with a grease pencil on the monitor screen works great).
I'm not sure of how you're planning on tilting your sample. Because you're using a TEM you may be best off tilting your beam instead of your sample. I've never tried this as our instrument wasn't able to do this, but it always seemed like a great way to go. I imagine there would be less refocusing problems. Less change in contrast too. Let me know how it works out if you try this method. There are two pretty good references on this technique you might want to check out. One is "The Perception and Measurement of Depth In the SEM", by A.Boyde in SEM 1979 Vol 2 page 67-78 (the part you want starts on pg 70). The other is "Introduction to Stereo Scanning Electron Microscopy" by Eric Chatfield in Vol 6 of Principles and Techniques of Scanning Electron Microscopy 1978. There are others I can give you if you want.
Another method you might try is horizontal displacement instead of tilting. The trick is to have enough displacement so as to produce sufficient parallax, but maintain enough similarity in the field of view that the brain can still fuse the two images. I didn't have much success with this technique. The above references describe this method as well.
Oh, heres a good one. While working on this little project of mine (which has now become somewhat of a recurring obsession) I discovered the National Stereoscopic Association (NSA). These people are very interested in any form of stereoimaging and would love to hear from you and help you in any way they can. They even have their own magazine Stereo World which is filled with you guessed it stereopairs. Its great if somewhat bizarre. And they have a cool website! Check it out at nsa-3d.org/nsa-membership.html and have your 3d glasses ready. You might try contacting Larry at larry-at-sapphire-star.com to see if he has any advice or suggestions.
Whew! I didn't realize I had so much to say! I hope this has been of some help. If you have any other questions or just want to discuss the frustrations of stereoimaging (and there are a few). Feel free to contact me at coopera-at-bigdog.engr.arizona.edu (the world's longest email address).
Good Luck and let me know how it turns out
Anne Marie Cooper
On Thu, 9 Apr 1998, Rick L Vaughn wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hey there imaging buffs } 1) Is there a way to create the red/green 3D images using stereo pair } TEM mages without buying an expensive 3D imaging program? } 2) What thickness of sections would be needed for TEM work to get } the effect of 3D depth? } 3) What is the best angle? I would be using a Philips 410LS } w/goniometer tilt stage. I'm going to hit the library but tips from the horses } mouth are always much better. Thanks in advance. } } Rick Vaughn } RLVAUGHN-at-MAIL.UNMC.EDU }
Cono Passione wrote: ================================================ I currently am trying to find a way to image composite fibers of 50 to 500 micron in length and 10 micron in width with out them crossing and touching.
The idea is to image, threshold, and then count and measure these fibers. right now we have to go through each field and manully measue them. This can be quite tedious and time consuming when there are thousands involved. We can use the image analyser to do fields automatically with some manual interaction.
Can anyone suggest a means of dispersing such fibers in the preperation process that will not allow them to touch and overlap??? We can also set the image analyser to randomly eliminate the ones that touch. This is an alternative but would like to be able to measure them all, if possible. Also this is done on just basic brightfield and does not require any special illumination such as polarized light. ================================================== This sounds like it could be an ideal application for SPI's "Tacky Dot Slides". If you are not familiar with this product line, you can get full details on our website given below.
You can chose between standard products with dots on either 500 or 1000 um "centers". If you went for the 500 um center to center dots, you could work with a dot size of only 15 um. That would ensure that only one fiber stuck per dot, but it would be theoretically possible for at least some small (but very small) population of fibers to be touching.
If you went for the 1000 um centers, the smallest dot size is 100 um. While you would be certain that no fibers on adjacent dots would ever be touching, there could be the possibility that there could be more than one fiber sticking on a given dot, giving another instance of fibers touching.
But despite these problems, it would seem that these "Tacky Dot Slide" products could either solve your problem completely or come very close to doing so. What you might need is a special order product with a smaller dot size on 1000 um centers.
Once mounted, the fibers could be imaged either by LM or SEM for the desired automated analysis.
Disclaimer: SPI Supplies manufactures Tacky Dot Slides (TM) as an exclusive licensee of E. I. DuPont de Nemours and Co., Inc. according to US Patent #5, 356,751.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
{smaller} I currently am trying to find a way to image composite fibers of 50 to 500 micron in length and 10 micron in width with out them crossing and touching.
{/smaller}
{smaller} The idea is to image, threshold, and then count and measure these fibers. right now we have to go through each field and manully measue them. This can be quite tedious and time consuming when there are thousands involved. We can use the image analyser to do fields automatically with some manual interaction.
{/smaller}
{smaller} Can anyone suggest a means of dispersing such fibers in the preperation process that will not allow them to touch and overlap??? We can also set the image analyser to randomly eliminate the ones that touch. This is an alternative but would like to be able to measure them all, if possible. Also this is done on just basic brightfield and does not require any special illumination such as polarized light.
{/smaller}
{smaller} Any suggestions can be sent to this address and I will reply. Thank You.
{/smaller}
{smaller} Cono Passione
{/smaller}
{/excerpt} { { { { { { { {
Dear Cono,
Other than an equally laborious process of aligning the fibers (you did not mention whether they were straight, curled, or crimped), one other approach is to find an image analysis system which has a special fiber module for analyzing fibers as they cross or touch. Companies to check on
I have students in my SEM course do an exercise in taking stereo pairs. It provides for a great show-and-tell at the end of the course. Since optimum angle depends on surface topography and magnification, I have different students do different angles and then c;compare end results. We usually use 7o as routine with 14o thrown in. The larger angle usually is overkill. It is very important to focus with Z-control and try to keep the image centered.
I have done a bit with TEM using a goniometer stage but not enough to give advise. It does work better if you have a rough surface specimen, either negative stain or sections in a type of resin that tears as you microtome (like lowicryls or LR White).
For reconstruction, I realign the images in photoshop, using the center of the image for alignment purposed. This is a critical step and must be done accurately for good final results. Using layers makes this routine. I put the original image (0 angle) in the first layer, cross-hairs on the second layer to mark my center point, and the second image on the third layer, etc. Reducing the transparency of the second image lets you easily see the cross-hairs and first image.
Using color channels, you can change the aligned images into your red and blue (or green) colors and adjust transparency to overlay them. However, it is easier to use an image analysis program such as NIH Image (free). I use ScanAlytic's program, IP Lab Spectrum which makes the conversion about a 20 second process.
The next problem is getting the final image into a form which can be shown to audiences. I have found that it is often difficult to get the right exposure using digital slide makers. The slides tend to come out too dark. You often also have color problems with some films. I have had the best luck just photographing the computer screen with a 35mm camera. I use Polaroid Presentation Film which gives you a "what you see is what you get" when photographing a computer screen. Kodak films are heavy on the blue in this instance and I haven't tried Fugi film (although it works great in some digital slide makers!). I use the option in the Adobe PhotoShop tool box of full screen mode which enlarges the image to maximum screen size and fills rest of monitor with a black border and then do an exposure series and get good slides every time.
Have fun with this...your audience will certainly appreciate it.
Debby Sherman, manager Microscopy Center in Agriculture Purdue University --------------------------------------
Hey there imaging buffs 1) Is there a way to create the red/green 3D images using stereo pair TEM mages without buying an expensive 3D imaging program? 2) What thickness of sections would be needed for TEM work to get the effect of 3D depth? 3) What is the best angle? I would be using a Philips 410LS w/goniometer tilt stage. I'm going to hit the library but tips from the horses mouth are always much better. Thanks in advance.
Rick Vaughn RLVAUGHN-at-MAIL.UNMC.EDU
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I am searching for a hardware/software solution to control access and provide accounting information for electron microscope use. We operate a multi-user academic facility with five microscopes and other analytical equipment. Presently, instrument access is controlled by custom written software running on a very old PC. Once a user has logged in, the PC switches the "ready" signal of an individual microscope using a serial I/O interface card. Unless a user has logged in, the instrument is disabled. Our existing system is becoming less reliable and is not easily upgraded.
Are other members of the microscope community using a similar approach? Does anyone know of a commercial software package for this type of application? The ability to work in a network environment (web-based perhaps) would be a plus.
I'm conducting research for a short paper and presentation on Scanning Electron Microscopy due 21 Apr 97. Any sources or information you would recommend would be greatly appreciated.
} At 06:02 PM 4/9/98 -0500, you wrote: } Hey there imaging buffs } 1) Is there a way to create the red/green 3D images using stereo pair } TEM mages without buying an expensive 3D imaging program? } 2) What thickness of sections would be needed for TEM work to get } the effect of 3D depth? } 3) What is the best angle? I would be using a Philips 410LS } w/goniometer tilt stage. I'm going to hit the library but tips from the } horses mouth are always much better. Thanks in advance. } } } Rick Vaughn } } RLVAUGHN-at-MAIL.UNMC.EDU } ----------------------------------------------------
Rick,
Responding to questions 2 & 3 above:
When I got my on-site training after instalation of our Philips CM12, I was handed a huge compendium of TEM applications tailored for the CM12. There is a section on stereo-TEM in there which I will copy and send to you by snail mail, if you'll send me your address.
Within it, is a table and a graph giving suggested tilt angles for stereo TEM as a funtion of section [or sample] thickness and magnification, which was worked out long ago by Dr. Lee Peachey. The trend is, the higher the magnification, the smaller the tilt angle between stereo pairs should be, and the thicker the sample, the smaller the tilt angle should be.
In response to question #1, so far I haven't played around with the red/green method of stereo presentation. I make a pair of black and white slides, by digitizing my TEM negs and use PowerPoint to print them out to a Polaroid slide maker. Then I use inexpensive stereo slide viewers ($7 ea.,from Reel 3-D Enterprises, www.stereoscopy.com/reel3d. I have no commercial interest in Reel 3-D.) to view the pairs, which works quite well. I'm presently working out viewing bacteria in 3D that have had immunogold labeling applied to their surfaces, to better localize the location of the gold.
Good luck!
Gib
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
I would be most interested in your findings. Please summarize them to the list. I have thought of using Microsoft's scheduler program to handle signing up, but it would not do the accounting, nor does it appear readily accessible over the web. I bet there would be a few takers for such a product.
At 11:01 AM 4/13/98 -0400, you wrote: } } I am searching for a hardware/software solution to control access and } provide accounting information for electron microscope use.
1. I would like feed back from anyone that has purchased or worked with a Gatan slow scan camera that was purchased in the last year. How is life after the salesman? Does the system measure up to expectations? Satisfaction with tech support? Software/ upgrade problems? Comparison of PC platform to MAC. Things that really need to be improved. Any other comments are also welcome.
2. We are also interested in a side mounted video camera. Comments & experiences are welcome on this subject as well.
I am in need of the manual for an LKB Knifemaker model 7801B. At some point I have lost the manual and now I need to overhaul this well-used device. I recall it was just a few pages long. Could someone take the time to fax it or scan it for me? It can be faxed to:
530 754 7536
If it is easier to FTP it, please email me for the FTP address and password.
Thanks in advance.
Rick A. Harris, Director Microscopy and Image Analysis Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 752 3085 fax raharris-at-ucdavis.edu
Well Kiddies my sputter coater died. And after all the help you guys gave me to fix the grounding problem too. Sheesh! Anywho, the question now is:
Is the vacuum evaporator good enough (or better?) to do some sputtering? We can tilt & rotate while doing it ;-O. Will this be sufficient for people doing low mag. work (not going over 1000X)?
I have to send the poor ole beastie away to be repaired, I think the Pirani gauge broke. The vacuum sucks but the gauge does not read out, it just lies there all the way to the left (but this is Berkeley so I guess that's fitting).
Promising not to riot while awaiting replies,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Is anybody using stereo pairs projected on a screen as a teaching tool? If so, would you mind describing how the images are lined up?
I 've thought for years that stereo projection had to be one of the most valuable assets to any classroom. I remember that the basics included 2 Ektamatic projectors with crossed sheets of polaroid across their lenses to allow viewers to use crossed polaroid glasses. A special screen was required, but I can't recall the details. I think I remember that one image was mounted as usual, and the other adjusted to the correct stereo location using a special guide that was invented by Lee Peachey, I think, and sold by one of the EM suppliers.
That was many years ago, so if there's a better technique now, please let us hear about it. And/or correct me if I'm wrong.
I am faced for the first time with having to process histological paraffin sections for TEM, and have a few questions. (First I should say this is mammalian tissue, and we must use histology first to locate the region of interest, which would require too many blocks to search for by TEM).
I've seen posts/literature in the past regarding the process of xylene treatment and rehydration before starting TEM prep. at the OsO4 stage. The two questions I have are: 1. Since we can have new sections cut from the original block, how thick should they be, and should they be collected onto slides treated with some type of mold release agent? Is there a better substrate than glass slides for this process?
2. If using glass slides, is there a way to create a "well" or chamber around the section so that I don't have to use whole Coplin jars full of OsO4, Spurr's resin, etc. during processing? I would much rather apply the reagents to the area of the section only, similar to applying immuno-reagents. However, I doubt that the hydrophobic pens we use would work with solvents and Spurr resin. Any ideas?
Any other tips from experienced individuals would be appreciated!
Thanks as always, Karen
-- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
Dear Paul, } } would you mind describing how the images are lined up? } We've used them for presentations in-house and at MSA. We've just put someone in the audience with stereo glasses, projected the images, and adjusted until the "audience" saw optimal stereo.
} A special screen was required, but I can't recall the details.
It is one coated with aluminum rather than glass. Metals do not change the polarization upon reflection, but glass does. I don't know where they can be obtained; we guard ours jealously. Yours, Bill Tivol
Dear Gib, } } Within it, is a table and a graph giving suggested tilt angles for stereo } TEM as a funtion of section [or sample] thickness and magnification, which } was worked out long ago by Dr. Lee Peachey. The trend is, the higher the } magnification, the smaller the tilt angle between stereo pairs should be, } and the thicker the sample, the smaller the tilt angle should be. } One thing to note is that this table gives easily seen stereo, but not accurate apparent z-values. Depending on what you're going to do with the info, you may or may not want to use the table. Most computer programs are capable of working out the correct z-values if you tell them the tilt angles. Yours, Bill Tivol
Dear LISTERS-FRIENDS, Thank your very much to everyone for your kind replies on my problems with electron miocroscope JEM-4000EX.
There was problem in logical vacuum system. Namely the chip No. 4B of VAC SYSTEM PB in VALVE UNIT had been foulted. That was replaced on new one. Now everything is OK. Again thank very much for help. Looking forward to help YOU in future.
Sincerely yours Anton Gutakovskii Laboratory of Electron Microscopy Institute of Semiconductor Physics Novosibirsk, Russia
Any kind souls out there who could advise on layout and specs for utility needs and vibration/EM field interferences for a lab space that would include light microscopy, infrared microscopy, SEM, XRD, HPLC, sample prep, and offices?
I will summarize written e-mail replies for the group. Drawings or plans can be faxed to 413/458-2314.
This is a multi-part message in MIME format. --------------7A44FA4AB075D78DF1B1F64F Content-Type: text/plain; charset=koi8-r Content-Transfer-Encoding: 7bit
Dear LISTERS-FRIENDS, Thank your very much to everyone for your kind replies on my problems with electron miocroscope JEM-4000EX.
There was problem in logical vacuum system. Namely the chip No. 4B of VAC SYSTEM PB in VALVE UNIT had been foulted. That was replaced on new one. Now everything is OK. Again thank very much for help. Looking forward to help YOU in future.
Sincerely yours Anton Gutakovskii Laboratory of Electron Microscopy Institute of Semiconductor Physics Novosibirsk, Russia
--------------7A44FA4AB075D78DF1B1F64F Content-Type: text/x-vcard; charset=koi8-r; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Anton Gutakovskii Content-Disposition: attachment; filename="vcard.vcf"
In our expanding microscopy core facility, we would like to "Gait" (sp?) cells using an instrument called the Cell-dyne whcich used laser scattering and some morphological features to identify the populations on a fixed slide. Is any one very familiar with the Meridian Ultima system, and do you know if these similar features are incorporated into their software and/or if cells can be not only morphologically identified, but also categorized as to laser scatter, size, shape, re-located with x+y precision, etc, automatically? Any leads would be help ful in this regard. Thank you, Tom Baginski
I wish to receive notice about the book "Manual of microscopic Analysis of Feedstuffs. 3rd Ed. The Amer. Assoc. of Feed Microscopists, 1992, p. 73-93" or the articles in the same book, autors BATES L.e coll. Feed ingredient descriptions of animal origin.
} } } {kszaruba-at-MMM.COM} 04/13 5:53 pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear All,
I am faced for the first time with having to process histological paraffin sections for TEM, and have a few questions. (First I should say this is mammalian tissue, and we must use histology first to locate the region of interest, which would require too many blocks to search for by TEM).
I've seen posts/literature in the past regarding the process of xylene treatment and rehydration before starting TEM prep. at the OsO4 stage.
The two questions I have are: 1. Since we can have new sections cut from the original block, how thick should they be, and should they be collected onto slides treated with some type of mold release agent? Is there a better substrate than glass slides for this process?
2. If using glass slides, is there a way to create a "well" or chamber around the section so that I don't have to use whole Coplin jars full of OsO4, Spurr's resin, etc. during processing? I would much rather apply the reagents to the area of the section only, similar to applying immuno-reagents. However, I doubt that the hydrophobic pens we use would work with solvents and Spurr resin. Any ideas?
Any other tips from experienced individuals would be appreciated!
Thanks as always, Karen
-- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
Hi Karen,
I do not mess with paraffin sections. Once an area of intrest is determined, the corresponding area from the paraffin block is cut out, cut up into small pieces than processed for EM. I start with a 1% OsO4 in Toluene for a minimium of 4 hours; wash 3X with acetone than infiltrate in 1:1 Spurr's:Acetone overnight. Embed in the morning into 100% Spurr's, polymerize and section. If you do have to work with paraffin sections only, have them cut as thick as possible (} 5 um) and process on the slides. This is the fun part: when ready to polymerize, fill up a beem capusle with resin, invert it over the section (area of intrest), clamp, tape, hold the capsule to the slide, and place in oven. Next day place slide on a hot hot plate and gently rock the capsule until it pops off the slide. ( I am successful about 50% of the time) Note: before the slide goes into the oven, wipe off the bottom and any area away from the area of intrest of resin so that only the section is covered with resin.
Best of Luck, Ed Calomeni Dept of Pathology Medical College of Ohio Toledo, OH 43614 ecalomeni-at-mco.edu
Hello again, World, [sorry for bothering again I mistyped the name in the former message.]
Looking for the address and fax number of the Company called Coherent Radiation, Inc. I just spent half an hour browsing, not to avail. So sorry for bothering you but I have no other way than asking. Thanks,
--Yves MANIETTE Universitat de Barcelona http://www2.gol.com/users/scscope/maniette/ENTREE.HTM
Looking for the address and fax number of the Company called Coherent Technology, Inc. I just spent half an hour browsing, not to avail. So sorry for bothering you but I have no other way than asking. Thanks,
--Yves MANIETTE Universitat de Barcelona http://www2.gol.com/users/scscope/maniette/ENTREE.HTM
Other than glass slides?? Yes permanox slides are available. They do not embed in Spurr and can be more easily separated from the section.
Small volumes of fixative?? Yes, try a small slide mailer that is used for shipping slides, I have used these with all solutions, with the exception of propylene oxide, we infiltrate these in alcohol+ resin. Some small slide chambers are also made for autoradiography, but are pricey.
Using glass slides?? I have had pretty good success separating sections or cells from glass slides by inverting the glass slide over the beam capsules, so the excess resin will drain away. And immediately after removing from the oven, set the glass slide on a block of dry ice. Wait until bottom of block is frosty and snap block off slide.
Have you tried some of the other resins available for light and TEM?? I can be contacted off the listserver if you have any other questions, and will be glad to help.. Marge
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
Don't know what materials you are evaporating or if it is a low or high vac system, but it should suffice for less than 1000x. Coating grain size is typically larger but you won't see it. I have found that with my low vac evaporator, film thickness is much more difficult to control and I don't use if for high mag/resolution work if possible. If you are coating with carbon (vs Au or AuPd) the stopping power is less, but again, at low mag it won't matter. Carbon will, however, yield a more noisy signal than Au since it tends to adsorb more incident beam and liberate less BSEs and SEs.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Well Kiddies my sputter coater died. And after all the help you guys gave me to fix the grounding problem too. Sheesh! Anywho, the question now is:
Is the vacuum evaporator good enough (or better?) to do some sputtering? We can tilt & rotate while doing it ;-O. Will this be sufficient for people doing low mag. work (not going over 1000X)?
I have to send the poor ole beastie away to be repaired, I think the Pirani gauge broke. The vacuum sucks but the gauge does not read out, it just lies there all the way to the left (but this is Berkeley so I guess that's fitting).
Promising not to riot while awaiting replies,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
by primemail1.pcom.net (8.8.5/8.8.7) with SMTP id MAA16551 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 14 Apr 1998 12:45:07 -0400 Message-ID: {35339273.216A-at-pcom.net}
i'm having trouble distingushing between the two are there any special etches i can use ?
Fellow list members, BibMic - A bibliography of books relating to Materials Microscopy, previously published on paper in Materials Characterization 36(1996)105 is now available on the net at http://bibmic.metalmat.ufrj.br
It lists over 1000 books, and is searchable by author, title and keywords.
Hope you find it useful Prof. Walter A. Mannheimer Dept. of Metallurgy and Materiais Eng. Federal University of Rio de Janeiro POBox 68505, 21945 Rio de Janeiro, Brazil Vox (55 21) 590-0579 Fax (55 21) 290-6626 wamann-at-metalmat.ufrj.br
} ---------- } From: Yves Maniette[SMTP:yves-at-giga.sct.ub.es] } Sent: Tuesday, April 14, 1998 8:10 AM } To: Microscopy List } Subject: Re: Coherent **Radiation**, INC.(Please forget about } previous msg) } =20 } = ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America=20 } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } = ---------------------------------------------------------------------- } -. } =20 } =20 } Hello again, World, =20 } [sorry for bothering again I mistyped the name in the former = message.] } =20 } Looking for the address and fax number of the Company called = Coherent } Radiation, Inc. I just spent half an hour browsing, not to avail. So } sorry for bothering you but I have no other way than asking. Thanks, } =20 } --Yves MANIETTE } Universitat de Barcelona } http://www2.gol.com/users/scscope/maniette/ENTREE.HTM } =20
Does anyone have an address, phone #, for a company called Genex Corp? Thanks for any help? Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children Dallas
I just know there is a association of microscopy in Internet. I work on the field for long time and I hope I can share the experience and knowledge with all of you. Please let me know if I need do something for joining , like fill a form.
Now I have a problem. I cut 50 micro meter paraffin section beautifully but I can not mount them well. There are a lot of wrinkles when the sections are dried. Maybe some experts can give me some idea. Thank you!
ou may want to be sure that your unit really isn't leak tight before sending anything away for repair. Can you put the gauge tube into an adapter (rubber stopper with a hole bored in it) and put this onto your mechanical pump and see if your gauge/pump combination works? If so then=
you have left out a sealing component while repairing the ground fault.
Keep smiling, it takes fewer electrons than frowning?
At last we have the printed course announcement for the RMC Materials Science Microtomy Course 1998.
This is the most comprehensive course available for those interested in doing polymers, metals, semiconductors, ceramics for TEM and SPM. =
The course dates are September 29-October 2, 1998 in Tucson,AZ. This is t= he fifth time the course has been held and is staffed by four experts in problem solving via ultramicrotmy and EM.
For complete details please see our web page; =
RMC-Scientific.com/microtomes/
Steve Miller Director of Sales, North America RMC 3450 S. Broadmont, Tucson, AZ 85713 Tel: 520-903-9366 Fax: 520-903-0132 Email: Steve.Miller-at-RMC-Scientific.com
} If you do have to work with paraffin sections only, have them cut as } thick as possible (} 5 um) and process on the slides. This is the fun } part: when ready to polymerize, fill up a beem capusle with resin, } invert it over the section (area of intrest), clamp, tape, hold the } capsule to the slide, and place in oven. Next day place slide on a } hot hot plate and gently rock the capsule until it pops off the slide. } ( I am successful about 50% of the time) Note: before the slide goes } into the oven, wipe off the bottom and any area away from the area of } intrest of resin so that only the section is covered with resin.
This is called the "pop-off" technique. We have posted a published paper delineating this technique in detail on our web site (address below). A second paper is available by snail mail.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
by hil-img-10.compuserve.com (8.8.6/8.8.6/2.10) id KAA15206 for microscopy-at-sparc5.microscopy.com; Wed, 15 Apr 1998 10:07:24 -0400 (EDT)
Giblab at GIBRALTAR STEEL wrote:
} I'm having trouble distingushing between the two are there any special } etches i can use ?
---------
After consulting with George Vander Voort (an authority on carbon steels and related etching techniques), I have discovered that there is not a good etchant to reveal retained austinite. An electrolytic method has be= en proposed in the past, but George told me he was never able to get good =
results from it. =
George's suggestion: =
Try Klemm's Reagent to tint etch the ferrite (if it's there). =
KLEMM's REAGENT: - Make up a stock solution of water saturated with Sodium Thiosulfate - Take 50ml of this stock solution and add 1gm Potassium Metabisulfite.
- Immerse sample (do NOT swab on the etchant). Watch the surface - of the steel, and remove from solution when the surface turns a - red/violet color. Rinse with water, ethanol and warm air dry.
Another option is to try Beraha's tint etchants listed on pg.643 of Georg= e Vander Voort's book, 'METALLOGRAPH Principles and Practice', published by McGraw Hill and available through ASM. This is not meant to be a book=
advertisement, but it is a handy reference if you are dealing with microstructural analysis on a regular basis.
Hope this helps. Scott D. Holt BUEHLER, LTD PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-6500 http://www.buehlerltd.com
I would like to receive information from anyone that has worked with a xenosput magnetron sputter coater. How is coating quality compared with conventional SEM coatings such as gold and chromium? Any other comments are also welcome. Thanks in advance.
Maria do Carmo Goncalves Institute of Chemistry University of Campinas Sao Paulo - Brasil e-mail: maria-at-iqm.unicamp.br
A position for a Senior Histology Technician is opening at Carolinas Medical Center in Charlotte, NC. Please address all inquiries to "Dr. Helen Gruber" {hgruber-at-carolinas.org} .
Thank you ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
I would like to receive information from anyone that has worked with a xenosput magnetron sputter coater. How is coating quality compared with conventional SEM coatings such as gold and chromium? Any other comments are also welcome. Thanks in advance.
Maria do Carmo Goncalves Institute of Chemistry University of Campinas Sao Paulo - Brazil FAX (55 19) 7883023 maria-at-iqm.unicamp.br
Without intent to re-open an old thread, I apologize to you all for comments made in haste and ignorance on this subject six months ago. I was visiting friends connected with Dartmouth last weekend, and was reminded of it by an article in the Dartmouth alumni magazine.
After reading the Aug. 97 Science article recommended by Robert Schoonhoven and some other references, including the glove tests showing astonishing permeability for normal lab gloves, I am now much more aware of how dangerous this stuff is compared to other mercury compounds. It's in the same league with VX nerve gas (which, in a ghoulish twist of bureaucratic humor, has an MSDS published by the Army).
Please forgive my erroneous pooh-poohing on this. It was a knee-jerk response to what looked at first like just another media scare story. I should know better by now than to react before having the facts.
With all this discussion of sterio pairs, I wondered if anyone was interested in a Wild M-5 sterio scope designed for viewing sterio pairs - scope has paired objectives that can be individually rotated to achieve convergence of sterio pairs for viewing as a 3-D image - I think it was designed for aerial photo and map viewing. I got it for a project I was doing a few years back and am done with it, so...... I am up for making someone a pretty good deal if you can use it.
Best to E-mail me at home spoefish-at-mindspring.com since the server here at work is often down for days (typical government stuff)
We use the 'pop-off' technique frequently in our lab (see Slap's post). If you must do it with sections, make sure they are 5 um thick or better. This will give you enough tissue to thin section.
We omit the OsO4 step, sense the ultrastructural morphology is very poor from formalin fixed paraffin embedded tissue.
(flame retardant: I understand some people have had good results using OSO4 with formalin fixed de-paraffinized tissue).
cya -gene
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
The Electron Microscopy Core Facility at the Mayo Clinic in Rochester, MN has an opening for an EM technologist to support both clinical and research projects. The laboratory offers expertise to collaborative projects which involve transmission and scanning electron microscopy. The laboratory is well equipped and has a history of excellent productivity and adequate funding. The successful candidate for this position will possess at least a bachelor's degree with experience in histology and/or electron microscopy. Additional experience in Immunology, Cell Biology, and Digital Imaging is desirable. Operating knowledge of transmission and scanning electron microscopes is preferred. The applicant must have excellent communicative skills and the ability to work well with a variety of personalities. The EM technologist interacts with all laboratory users in order to accomplish specific research and clinical goals with respect to electron microscopy procedures. Duties include: all aspects of specimen preparation for a variety of biomedical samples for TEM and SEM; operation of TEM and SEM; negative developing and printing; digital image capture, processing and archiving; and reporting. The technologist will also perform advanced research procedures including immunoelectron microscopy, x-ray microanalysis, and microwave processing. Mayo offers a competitive salary and benefits package. Candidates must be legally authorized to work in the United States. If interested please submit a cover letter and resume referencing job posting #98-818.col to:
Kaine A. Kerkhoff Mayo Medical Center Human Resources-OE 1 Rochester, MN 55905 Fax: 507-284-1445 Email: kerkhoff.kaine-at-mayo.edu
For more information about Mayo visit our homepage at: http://www.mayo.edu.
Jon Charlesworth, Coordinator Electron Microscopy Core Facility Mayo Clinic 1426 Guggenheim Building Rochester, MN 55905 ph: (507) 284-3148 fax: (507) 284-9349 email: charlesworth.jon-at-mayo.edu
Quick question.....has anyone ever experienced poor resin infiltration of tissue culture cells grown and processed in Permanox plates. I have performed this procedure a number of times with no problems and today, after looking at the sections on the TEM, the cells appear condensed and poorly infiltrated. I would be thankful for any shared experiences and solutions. Because this is such a specific situation, feel free to respond to me at my e-mail address. Thank You!
Sandy Perkins
Laboratory for Neurotoxicity Studies VA-MD Regional College of Vet. Med. VA Tech
We just got our Fuji Pictrography 3000 and the prints are flying out the door. I can't believe the quality. My question is: Does anybody have a great supplier (in regards to price) for the paper and donor material. I am paying $280 for the donor and $68 for a roll of glossy paper. Thanks. Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
At 11:30 15/04/98 -0300, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It is a vital part of our extensive high resolution scanning EM work where we hope to obtain featureless coating of insulating specimens.
The coating quality obtained with the xenosput sputtering chromium is as smooth as we can currently obtain with any technology. By comparison, gold sputtered in the conventional way is extremely rough. The only method which comes close is to sputter platinum at very low currents and pressures with a conventional magnetron sputter head.
Of course chromium CAN be sputtered in any vacuum system, but it will have a high content of chromium oxide due to residual gas. The Xenosput has the cleanest imaginable vacuum system in that a pre-coat sputter of titanium in the work chamber purges all gases except the xenon from the work chamber atmosphere. So you can apply a coat of pure metallic chromium which will not oxidise for several days.
The xenon is used as it has better characteristics for sputtering. Under xenon energetic neutral bombardment of the sputtered film is minimised and heat buid up in the specimen is reduced.
Hello All, I was wondering if anyone has had any problems with hardening of tissue when using lanthanum nitrate as an extracellular tracer? Also, is it advisable to use lanthanum in all processing solutions ie through osmium fixation, dehydration etc, or only in the initial incubation buffer? Any advice would be much appreciated.
Thanks in advance
--------------------------- Miss Peta Clode Zoology Department LaTrobe University Bundoora, Victoria Australia. 3083.
Has anyone come across an adhesive called " M Glue" ? It used to be = available 6-7 years ago and was superb for mounting small powders. I = think it was a water based adhesive which was "milky" at first and = cleared after 20 mins to leave a tacky surface which gave excellent = adherence and stability. We used to purchase it from the UK but I = think it was manufactured in S.Africa. We have been unable to find a = suitable replacement. Does anyone know of a similar product ?
Thanks,
Colin Reid
Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#c8e0d8} {DIV} {FONT color=3D#000000 size=3D2} Has anyone come across an adhesive = called "=20 M Glue" ? It used to be available 6-7 years ago and was = superb=20 for mounting small powders. I think it was a water based = adhesive=20 which was "milky" at first and cleared after 20 mins to leave = a tacky=20 surface which gave excellent adherence and stability. We = used to=20 purchase it from the UK but I think it was manufactured in = S.Africa. =20 We have been unable to find a suitable replacement. Does = anyone know=20 of a similar product ? {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Thanks, {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Colin Reid {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {BR} Electron Microscope = Unit, {BR} Trinity College=20 Dublin, {BR} Dublin 2, {BR} Rep. of Ireland. {BR} Tel: 353-1-6081820 {BR} Fax:=20 353-1-6770438 {BR} email: {A=20 href=3D"mailto:creid-at-tcd.ie"} creid-at-tcd.ie {/A} {/FONT} {/DIV} {DIV} {/DIV} {/BODY} {/HTML}
I have been asked to calculate the mfp (elastic and inelastic) of electrons in wool with increasing levels of a contaminant. At this stage I am going to consider the wool as a carbon then add in different levels of the contaminant. In the first cases this contaminant will be sulphur and bromine.
Is there any shareware out there (or even a macro or for something like Excel) which could perform these calculations and display the separate mfps. I only have access to PC's and Macs and can't get access to any compilers so the code would need to be executable!! I have seen the NTLAMBDA program in the archives and it is just what I want, but I have no way of compiling it!!
Any help would be greatly appreciated.
Colin Veitch
Instrumentation Scientist CSIRO Division of Wool Technology PO Box 21, BELMONT, Vic. 3216. Australia.
We are trying to find, to purchase or trade, working or not working, a LVC Sputter Coater once manufactured by Plasma Sciences, Inc. up until a few years ago. We believe that some of these units also could have carried the name of Energy Beam Sciences, Inc.
Please contact me off line if you have such a system that is excess to your current needs.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We have tried Temp-Fix but our powders are both temperature and solvent sensitive. At the moment we use "Sticky-Tabs" which do not give good stability. I am hoping that someone will know more about the original "M-Glue" to try and source a new supply. As with a lot of EM products, such as resins, they are used for other functions and the EM use is just an off-shoot. I know the company in the UK have tried to source the material and failed.
Colin Reid Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
-----Original Message-----
Colin
NTLamdba will be running as a WWW applet in about a week. I'll shift around my "order of things" to convert and try to help you out. You will need a Java aware browser like Netscape The URL is:
http://tpm.amc.anl.gov/NJZTools
Just login next week and look for the EELS MFP Link to be activated.
} Hi All, } } I have been asked to calculate the mfp (elastic and inelastic) of } electrons in wool with increasing levels of a contaminant. } I have seen the } NTLAMBDA program in the archives and it is just what I want, but I have } no way of compiling it!! } } Colin Veitch
Is SEM considered radiation creating instrument that has to be registered? ******************************************************************* Pnina Ari-Gur, D.Sc. Materials Science and Engineering Western Michigan University Office: (616) 387-3372 Kalamazoo, MI 49008 FAX: (616) 387-6517 email: arigurp-at-wmich.edu or pnina.ari-gur-at-wmich.edu also: arigur-at-lab2.cc.wmich.edu *******************************************************************
In the state of Michigan all electron microscopes are considered radiation producing devices. They must be registered and readings taken once per year. There must be a registration tag on each instrument as well as the current inspection certificate. The Michigan Dept. of Consumer and Industry Services takes care of this. However, the registration and inspection is usually coordinated by a radiation safety officer at each university. I suggest you make a few calls at your university to get the complete story. The State can levy fines if you are not in compliance; however they are usually lenient, especially if you were not aware of the law. Stanley L. Flegler, Assistant Director Center for Electron Optics Michigan State University flegler-at-pilot.msu.edu
We are in the market for a dye-sub printer - primarily for SEM images, Electron Probe Maps, TEM images, transparencies, .... The Sony UP-D8800 was recommended but I would appreciate comments on other printers concerning the print quality, reliability, etc.
Thanks, steve rozeveld
} Steve Rozeveld } } Dow Chemical Co. } 1897 Bld. Door E-43 } Midland, MI 48667 } sjrozeveld-at-dow.com } % (517) 636-5167 } Fax: (517) 638-6443
Colin Reid wrote: =============================================== We have tried Temp-Fix but our powders are both temperature and solvent sensitive. At the moment we use "Sticky-Tabs" which do not give good stability. I am hoping that someone will know more about the original "M- Glue" to try and source a new supply. As with a lot of EM products, such as resins, they are used for other functions and the EM use is just an off- shoot. I know the company in the UK have tried to source the material and failed. ================================================ You might be thinking about "M-Bond 610". If that is indeed the one you are talking about, and it is used in several different ways around EM labs, you can find it on our website listed below. Its original use was indeed outside of EM, it was developed for the mounting of micro-strain gages on surfaces.
But for temperature and solvent sensitive powders, have you considered our Tacky Dot (TM) slides? This will enable you to mount your powders without exposure to solvents or temperature and the particles end up on orthogonal centers for easy viewing and analysis. These two are described on our website below. The advantage of the Tacky Dot slides over even M-Bond 610 is that there is no danger that your particles of interest will "sink into" the liquid adhesive while it is polymerizing. After all, even the M-Bond 610 has vapors and if your powders are indeed solvent sensitive, then they would or could be sensitive to the vapors from such solvents as well.
Disclaimer: SPI Supplies offers both types of products, as well as Temfix (TM), so we would have an obvious interest in promoting the use of these items.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The Midwest Microscopy and Microanalysis Society Materials Science Meet= ing will be held on May 22, 1998, hosted by Dr. Nigel Browning of the Depar= tment of Physics, University of Illinois at Chicago. In addition to a spea= ker slate of internationally renowned scientists, the meeting will showcase= the new electron microscopy lab at UIC with a tour of the facility followin= g the scientific sessions.
Invited presentations will cover the following topics: phase contrast imaging in TEM, scanning tunneling microscopy, electron energy loss spectroscopy, enery dispersive x-ray spectroscopy, and other surface analytical microscopic techniques.
The program will begin with an introduction at 8:45 a.m. and the first speaker at 9:00 a.m. There will be poster sessions and a buffet lunch.= The last scientific session will end at 4:00 p.m., with the facility tour following this session.
MMMS members will receive a mailing in the next few days that will incl= ude the meeting agenda and a map with travel information. Anyone else requ= iring information may contact me at (847) 935-0104 or by e-mail (preferred) = at jane.a.fagerland-at-abbott.com.
Steve asks: } ... } } } We are in the market for a dye-sub printer - primarily for SEM images, } Electron Probe Maps, TEM images, transparencies, .... } The Sony UP-D8800 was recommended but I would appreciate comments on } other printers concerning the print quality, reliability, etc. } } ...
You probably should have let us know with which computer you'll interface with ... much of what you're asking has as much to do with the software ... However, we use our dye-sub withing a situation of many different platforms all connected via TCPIP. We went with the Codonics NP1600 networking printer and haven't looked back. But if I were to wish for perfection, it would be that it printed with a CMYK ribbon rather than to create grayscale images with CMY. I don't know that any dye-sub will print grayscale without a tint if their ribbon doesn't have the K transfer. The only dye-sub I'm aware of with includes K with CMY is Tektronix ... I'd like to know of others. (... BTW ... All dye-subs will offer a K ribbon ... but switching between color and grayscale printing is not practical ...)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
There is a product called Microstik offered by Ted Pella (Cat. #16033) which sounds as if it may do what you need. I have only used it a couple times, but as I recall it did a good job of adhering fine particulates without drowning them.
No financial interest, etc.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
} But if I were to wish for perfection, it would be that it printed with } a CMYK ribbon rather than to create grayscale images with CMY. I don't } know that any dye-sub will print grayscale without a tint if their } ribbon doesn't have the K transfer. The only dye-sub I'm aware of with } includes K with CMY is Tektronix ... I'd like to know of others. (... } BTW ... All dye-subs will offer a K ribbon ... but switching between } color and grayscale printing is not practical ...)
One addendum to Michael Shaffer's comments:
We also are extremely well pleased with our Codonics 1660M dye sub printer.
We are using the Chroma Vista color dye sub media (CMY) and the b/w media for gray scale printing. They are supposed to be releasing a Direct Thermal media for b/w that does not use a transfer ribbon. Perhaps at that juncture, one could leave the color ribbon in place and use the Direct Vista paper for b/w printing since the medium is in the paper. Of course, this would require shutting off the color ribbon somehow. Nifty idea if they could pull it off. Time will tell.
JB
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Thanks to everyone who responded to my question! There were a lot of good suggestions, unfortunately it will be another couple weeks before I get to try them out.
Some of the suggestions were: 1. Use the paraffin tissue block itself instead of sections. [This is not an option on these samples, but will keep in mind for future.]
2. Make sure the sections are } 5 microns thick, as much as 20 microns.
3. Collect on: superfrost plus glass slides, Permanox slides, or ACLAR.
4. Or, process free-floating sections in vials (sections should be very thick and may have curling problems).
5. To reduce reagent volumes, add drops on top of section and cover with parafilm, or invert section over drops using toothpicks to prop up slide. Or, process in slide mailers, which are smaller than Coplin jars.
Thanks again, Karen -- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
I would greatly appreciate any information as to the possible health hazard of using halogen lighting as a light source in microscopy. Having scant resources at hand I built a light source using halogen bulbs. I was told that they are a source of UV radiation. My question is should this be of any concern?
Stanley L. Flegler responds: } } } ... } ... In terms of testing, they normally ask that it be tested at its } normal opering voltage. Stanley L. Flegler }
I figured the primary interest would be x-rays, but was looking to separate SEMs from TEMs because "normal operating voltages" are generally less than 30kV, rather than generally more than 100kV. Is there any rationale for claiming 30keV x-rays simply cannot escape the instrument???
Can anyone direct me to a source of those nifty lights/signs that go on the outside of a darkroom door with a switch inside to turn them on and off? I have looked several places with no luck. Even simple home made ideas would work for us.
Thanks
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Pnina Ari-Gur wrote: ================================================ Is SEM considered radiation creating instrument that has to be registered? ******************************************************************* This is a legal kind of question and the answer depends on the political jurisdiction in which you reside. In the USA, it is determined on a state- by-state basis, and I suspect in other countries, on a country by country basis.
In Pennsylvania, we are required to register each instrument with the state. We also are required to have someone radiation check each registered instrument. The requirements for NJ are different than they are for PA. So you have to contact the officials in your own state, usually located in some kind of a "radiation physics" kind of department, and find out about the legal requirements for your own specific situation.
But these requirements should not be lightly dismissed as some kind of bureaucratic boondoggle. There have been enough "horror stories" over the years and these are the kinds of stories that literally "drive" regulators and the promulgation of regulations:
a) On several occasions, a TEM "viewing glass" was broken, and when hearing the high price of a replacement from the manufacturer (which would have been lead glass), the user replaced the glass (without realizing what they were doing) with more ordinary silica glass, resulting in long term radiation exposure to the operator.
b) Some of the older instruments designed in the 1950's at a time before the hazards of radiation exposure were fully appreciated became recognized as "known emitters". The affected manufacturers apparently did various retrofits to those instruments once the hazards became known. I can myself remember operating one of these instruments but only while wearing a lead apron! But I would still be concerned about any pre-1960 instrument that has not been recently and regularly checked for radiation leakage, including the adjustment knobs on the lenses.
c) I have myself encountered at least on one occasion an SEM that had been modified with the placement of a "see through" port (to observe cathodoluminesence) which was not made of lead glass. In another instance, my admonitions caused an SEM user to not install a "port" plate with "Plexiglas".
Modern instruments seem to be made in a way that x-rays just can not escape during regular operation. And if the instrument is so far out of alignment that x-rays do escape, then it would also be out of alignment to the extent that it would not be possible to hold a high vacuum and therefore even get high voltage and a beam. So the end result is again, there is no chance of escape of x-rays.
Radiation badges seem to give some peace of mind with regard to the kinds of massive kinds of leaks described above. However, the leaks that might be occurring, such as when using adjustment knobs on the column, and which would result in exposure to the fingers and hands, would not be registering typically on a radiation badge. Hence one has to guard against the radiation badge becoming a false sense of security. And one should pay particular attention to radiation that might be emanating from the adjustment knobs on a column.
My firm offers no products or services in the radiation physics area. These are my own views and opinions as a result of 35 years in the world of EM. I am told by some that I over exaggerate the hazards of a modern SEM or TEM. But not everyone uses modern EMs and also, any laboratory is vulnerable to someone doing something dumb, like replacing a broken glass window with something other than the manufacturer's recommended replacement part (and without anyone even knowing about it). That is why I believe that any EM instrument, whether required by law or not, be checked periodically for radiation leakage, just to be sure.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The cheap (!) FARGO thermosub printers also print with CMY, CMY+Overlay (same ribbon) OR with a CMYK ribbon OR with a K ribbon (b/w). Resolution 300x600 max. We own one, it works well, but I don't have possibility to compare it with others, so can't state anything about that. We bought it because of tests in computer journals (don't remember which) and our limited money.
you wrote:
} But if I were to wish for perfection, it would be that it printed with } a CMYK ribbon rather than to create grayscale images with CMY. I don't } know that any dye-sub will print grayscale without a tint if their } ribbon doesn't have the K transfer. The only dye-sub I'm aware of with } includes K with CMY is Tektronix ... I'd like to know of others. (... } BTW ... All dye-subs will offer a K ribbon ... but switching between } color and grayscale printing is not practical ...)
Dr. Arthur Schuessler University of Heidelberg Zellenlehre Im Neuenheimer Feld 230 D-69120 Heidelberg Germany
We were inspected by the National Radiological Protection Board (our official Radtiation Advisers, in UK) who found nothing of concern but started us wondering. For about two years thereafter, we wore radiation monitoring badges initially which showed nothing. Then we posted badges at strategic place around the column (held on by sticky tape). We never found anything on them, although sensitive radiation counters could occasionally pick up counts from certain areas (such as around where the x-ray detectors enter the column!). The worst problem was the plaster (skim coating) on the wall of the room, the natural background in the building is relatively much higher!.
I would not think that you have any real problem. These bulbs are used generally in microscopy, slide projectors, car headlights and probably other uses too. All lamps will produce a spectrum of radiations but not excessively in the UV band unless designed to do so specifically, like high pressure mercury lamps for fluorescence microscopy. Most simple light microscopes don't incorporate UV filtering in their setup.
Common sense says don't spend long looking at it i.e. don't expose your eyes directly to the light!
I would be more concerned about the electrical safety. You need to make sure the unit checks out ok in that area. Don't risk electrocution!
In many states, it is. The funny thing is that the 'soft' radiation emitted by an SEM (should there be a breach in the shielding) is often not detectable by the survey equipment required. In most cases, you'll get higher readings from an EDS monitor.
As I recall (a fragile thing), Michigan is indeed a state that requires such testing. Your institution can likely provide the certification through their plant maintenance department.
TEM operators should be more attentive to these requirements. The higher operating voltages produce 'harder' radiation that can be more penetrating. In addition, there are more opportunities for emission through the column and chamber accesses.
Brings to mind a TEM manufacturer who, many years ago, supplied instruments with viewport glass that was not leaded per spec. Many old time TEM operators took to wearing their radiation badges after that.
} } Is SEM considered radiation creating instrument that has to be } registered? } ******************************************************************* } Pnina Ari-Gur, D.Sc. } Materials Science and Engineering } Western Michigan University Office: (616) 387-3372 } Kalamazoo, MI 49008 FAX: (616) 387-6517 } email: arigurp-at-wmich.edu or pnina.ari-gur-at-wmich.edu } also: arigur-at-lab2.cc.wmich.edu } ******************************************************************* } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Chuck made some interesting comments on radiation leaks from EM's. I am sure we have all come across very dangerous situations over the years. I can remember my first job in a hospital pathology lab where the Osmium was mixed on an open bench because the main operator had destroyed his sense of smell. Thankfully I moved on quickly !
There was one potential source of X-Rays that was not mentioned. With side arm insertion on TEM's if the arm is left out the beam may be switched on for alignment etc. ). This produces X-Rays at eye level. ( We use this to test our Geiger counter ! ) One to watch.
Colin Reid Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
However, the leaks that might be occurring, such as when using adjustment knobs on the column, and which would result in exposure to the fingers and hands, would not be registering typically on a radiation badge. Hence one has to guard against the radiation badge becoming a false sense of security. And one should pay particular attention to radiation that might be emanating from the adjustment knobs on a column. {snip} --------------------------------------------------------------- If such a concern exists, TLD* finger rings are available.... *- ThermoLuminescent Dosimeter
I use them from time to time. Not to measure dose from my SEM. but to check the dose from some of the samples I must prepare!
Please allow me to inform you of a Minisymposium on SEMICONDUCTOR RADIATION DETECTORS which will be held during the 34th Conference on Microelectronics, Devices and Materials (MIDEM) in Rogaska Slatina, Slovenia, September 23-25, 1998.
In the framework of the minisymposia, several invited speakers will present different aspects of the chosen topic, thus offering the audience valuable complete information. The minisymposium invited speakers will be: Dr. Gerhard Lutz from Max-Plank Institute fur Physic, Munich, Germany; Dr. Joseph Kemmer, KETTEK Gmbh, Germany, Dr. Peter Weilheimer, CERN, Switzerland; Dr. Walter Bonvicini, Instituto Nazionale di Fisica Nucleare, Trieste,Italy
Complete information regarding submission of papers, topics of invited lectures, conference location etc. can be found on Web page http://pollux.fer.uni-lj.si/midem/conf98.htm
(Deadline for submission of papers is May 15th)
Hope to see you in Rogaska Slatina!
With best regards, Dejan Krizaj
*********************************** Dejan Krizaj, Ph.D. Laboratory for Electron Devices Faculty of Electrical Engineering University of Ljubljana Trzaska 25 1000 Ljubljana SLOVENIA
Email: dejank-at-fe.uni-lj.si ************************************ Disclaimer: This email was sent to addresses obtained through various sources. We apologize if you or your institution is not involved in activities covered by the Conference and Minisymposium. If, however, you know for persons that might be interested in this information, please forward them this message.**********************************
Please allow me to inform you of a Minisymposium on SEMICONDUCTOR RADIATION DETECTORS which will be held during the 34th Conference on Microelectronics, Devices and Materials (MIDEM) in Rogaska Slatina, Slovenia, September 23-25, 1998.
In the framework of the minisymposia, several invited speakers will present different aspects of the chosen topic, thus offering the audience valuable complete information. The minisymposium invited speakers will be: Dr. Gerhard Lutz from Max-Plank Institute fur Physic, Munich, Germany; Dr. Joseph Kemmer, KETTEK Gmbh, Germany, Dr. Peter Weilheimer, CERN, Switzerland; Dr. Walter Bonvicini, Instituto Nazionale di Fisica Nucleare, Trieste,Italy
Complete information regarding submission of papers, topics of invited lectures, conference location etc. can be found on Web page http://pollux.fer.uni-lj.si/midem/conf98.htm
(Deadline for submission of papers is May 15th)
Hope to see you in Rogaska Slatina!
With best regards, Dejan Krizaj
*********************************** Dejan Krizaj, Ph.D. Laboratory for Electron Devices Faculty of Electrical Engineering University of Ljubljana Trzaska 25 1000 Ljubljana SLOVENIA
Email: dejank-at-fe.uni-lj.si ************************************ Disclaimer: This email was sent to addresses obtained through various sources. We apologize if you or your institution is not involved in activities covered by the Conference and Minisymposium. If, however, you know for persons that might be interested in this information, please forward them this message.**********************************
Look into the Codonics 1600 and 1660. Quality is excellent, company support is great and cost per print is certainly comperable if not less than many other brands. It is built using a Kodak engine but has the most advanced software of any printer that I have heard of in this price catagory ($9000-13,000). This printer permits adjustment of gamma and contrast at the printer....a great feature when printing from drawing programs which do not permit these changes. We do a lot of labeling of images and gels in a drawing program and want to print directly with ability to adjust brightness and contrast of image if necessary.
Also it is very easy to send multiple files to a single sheet of paper with the printer doing the scaling to fit. You can instruct the printer to divid a sheet into 4-6 or more sections and then send that number of images to it. This is great for making quick hard copies of digital SEM images at low cost.
I have requested a number of feature updates since we purchased our printer 4 months ago and the company has been very receptive to addressing these issues. And if there is a mechanical problem with a printer that cannot be fixed over the phone, they are not opposed to drop shipping a new printer so your down time is minimal.
Debby Sherman, manager Microscopy Center in Agriculture Purdue University
--------------------------------------
We are in the market for a dye-sub printer - primarily for SEM images, Electron Probe Maps, TEM images, transparencies, .... The Sony UP-D8800 was recommended but I would appreciate comments on other printers concerning the print quality, reliability, etc.
Thanks, steve rozeveld
} Steve Rozeveld } } Dow Chemical Co. } 1897 Bld. Door E-43 } Midland, MI 48667 } sjrozeveld-at-dow.com } % (517) 636-5167 } Fax: (517) 638-6443
P.S. My apologies if I am covering an old topic.
}
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This came to me this morning via EduCom's edupage technology digest. Since CD-ROMs are most folks choice for archiving data and digital images, I thought this would be appropriate to pass on. Doug
DIGITAL ISN'T FOREVER "Digital information lasts forever, or five years -- whichever comes first," says a senior computer scientist at RAND Corp. The problem is that computer experts are finding out that under less-than-optimal conditions, digital tapes and disks, including CD-ROMs, can deteriorate in as little as five to 10 years. And the decay, although it happens gradually, isn't evident until it's too late, says the founder of Voyager Co., which makes commercial CD-ROM books and games. "CDs have a tendency to degrade much faster than anybody, at least in the companies that make them, is willing to predict." At the same time, as data is ported from an antiquated platform to a newer system, often there are bits that fail to make the transition. Sometimes it's just a matter of footnotes disappearing, but sometimes whole categories of data are lost. "It's like playing the child's game of Telephone. It doesn't take many translations from one media to another before you have lost significant aspects of the original data." (Business Week 20 Apr 98) ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
1. V1 doesn't work in auto mode -Control circuit is malfunction i.e. IC1(vac control),IC8, Tr5. V1 won't work in manual. -check V1 and solinoid valve, air pressure 2. if the control circuit and V1 working properly at 190 UA V1 should be open to get HVAC. 3. All valve will operate manually while they are running in Auto mode because we by pass the control circuit .
4. V5 doestn't operate in auto
-check airlock door swich
regards,
Paiboon Nuannin ---------------------------------------------------------------------------- Director Scientific Equipment Center Prince of Songkla University Hatyai 90112 Thailand
} } } ---------- Forwarded message ---------- } Date: Mon, 23 Mar 1998 10:03:47 -0400 } From: Brian McIntyre {mcintyre-at-optics.rochester.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: JEOL35 vacuum problem folowup } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } hi- } } thanks to all who sent info on about testing the vacuum system. } } seems i may have more problems: } } 1) the leak rate from HVac is {3uA/min } atm=250uA } Hvac=25uA on vacuum meter test points (OK) } } 2) V1 doesn't work in auto mode, and many times won't work in manual } } 3) DP heater warms up (light on) in about 20minutes } } 4) R4 vacuum control pot will not make V1 open when vacuum meter {190uA } while in auto mode } } 5) solenoid valve for the closing side of V1 will not release pressure } so that the opening side of the valve (when pressurized) will force the } valve to open.... most of the time (related to #2 above) } } 6) some of the valves (like v4) will actuate manually when the } controls are } in the auto mode (is this normal...or an indication that the logic is } all messed up?) } } 7) V5 (airlock) will not open in auto mode, but does in manual (as } does the } associated leak valve) } } i'm really confused about this since it seems to be a moving target with } many appearent "failures". is there one circumstance or condition that } would explain these observations???? } } thx in advance! } brian } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627 } } 716-275-3058 } 716-244-4936(fax) } } } }
At 12:32 17/04/1998 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to know what is your EM. As a field engineer since 22 years, I can tell you NO SEM and NO TEM from any manufacturer has Xray leakage The problem is coming and at that point I agree with Mr Garber, when the people has modified or replacing original parts with local supply for example. Speaking about the microscope I know, Jeol, It's for example impossible to generate beam on a 2010 (200kv) with no sample holder, there is an opto coupler detector. Of course if you insert something that simulate the holder you can have the beam. But this is a wrong manipulation. Like to remove a Gamma source from it's lead castle. You can do it but you have to think about that because you have be warned! Same for electron microscopy. May be you can check the statistics about cancer in the EM environment, for myself since all that years, all my collegues and competitors are still living and I'm sure that cigarettes cause more painfull than EM radiation. This reply is not for starting a polemic but just to tell that ,we, manufacturers of microscopes are responsible people and it's also our health when we are working on theses machines. Cheers.
=================================== Jacky Larnould JEOL Europe SA Service Manager mailto:larnould-at-worlnet.fr mailto:larnould-at-mnet.fr WEB: {http://www.jeol.com} Phone:33 (0)4 67 72 28 26 Fax :33 (0)4 67 79 54 90
I am looking to purchase a video camera that can be mounted on a light microscope to display microscope sections via a video projector. The video projector is a Sharp model XGE1200U. The resolution of this projector is 800X600 for PC or 832X624 for Mac. I want to be able to project real time pictures from a light microscope using this same system. Last fall I tried this with an older video camera mounted on this microscope (640 x 480 resolution) but the images were too pixilated to be really useful. However, images projected from a Macintosh computer were fine. Does anyone know of a high line resolution camera (video rate) that they have used for a similar purpose that might help me out?
Kenneth A. Taylor, Ph.D. Office phone: 850-644-3357 Institute of Molecular Biophysics Lab phone: 850-644-4104 Florida State University EM room phone: 850-644-8769 Tallahassee, FL 32306-4380 Fax: 850-561-1406 E-mail: taylor-at-bio.fsu.edu Home pages: http://www.sb.fsu.edu/~taylor/ http://www.fsu.edu/~biology/faculty/Taylor/kat.html
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Leica, the first name in ultramicrotomy, Diatome US, and Electron Microscopy Sciences announce another in a series "ultramicrotomy of materials" workshops.
Bring Your Own Samples!!!
Focus on hands-on participation of the following:
Embedding of industrial samples Specimen trimming
Ultramicrotomy of hard materials SEM applications of microtomy
Ultramicrotomy of polymers Section collection & handling
Low temperature sectioning
Course Instructors:
Dr. Tom Malis Mr. Helmut Gnagi Materials Characterization Group Leader Hard Materials Specialist Canadian Federal Laboratory Diatome, Ltd. Switzerland
Mrs. Ani Issaian, MS Ms. Kathy Johnson California Inst. of Technology Materials Analyst-SEM Polymer Scientist Gates Rubber Co.-Denver
Where: University of Colorado, High Voltage EM Facility Boulder, CO
When: May 19-21, 1998
Tuition: $1,100.00 USD
For more information contact:
Paul DeGeorge 800-248-0665 X2279 Diatome US 215-646-1478
id sma001490; Fri Apr 17 14:08:01 1998 Received: from elba.wadsworth.org (elba [172.16.1.100]) by newton.wadsworth.org (8.8.6.Beta3/8.7.1) with ESMTP id OAA27471; Fri, 17 Apr 1998 14:01:50 -0400 (EDT)
Dear Jacky, } } I would like to know what is your EM. } As a field engineer since 22 years, I can tell you NO SEM and NO TEM from } any manufacturer has Xray leakage } The problem is coming and at that point I agree with Mr Garber, when the } people has modified or replacing original parts with local supply for example.
Here is where Charles Garber's points can be particularly well-taken. We had an old JEOL JEM200 (no longer in service) which had a radiation leak at the side-entry airlock. This leak was very directional, and with the operator seated, there was no exposure. However, if the operator were to stand up with the beam on, (s)he would, indeed, be exposed to x-rays. This occurred with the original equipment--there were no modifications. It must be added that this scope was manufactured before ANSI standards for radia- tion-producing equipment were in effect, and finding this sort of leak was part of the impetus for developing those standards.
} Speaking about the microscope I know, Jeol, It's for example impossible to } generate beam on a 2010 (200kv) with no } sample holder, there is an opto coupler detector. Of course if you insert } something that simulate the holder } you can have the beam. But this is a wrong manipulation. Like to remove a } Gamma source from it's lead castle.
This is a good example of an improvement of the safety of EM's. However, the old instrument would leak x-rays whether a specimen was in or not.
} You can do it but you have to think about that because you have be warned! } Same for electron microscopy.
Yes. There's only so much that the manufacturer can do. One must take responsibility for one's own safety. (Of course, someone must provide appropriate safety instructions, but it is up to the individual to follow them.)
} May be you can check the statistics about cancer in the EM environment, for } myself since all that years, all my } collegues and competitors are still living and I'm sure that cigarettes } cause more painfull than EM radiation.
Even if the EM produces some radiation, there is likely to be a larger effect from concrete walls (potassium-40) or living at altitude, which increases radiation exposure 24 hours a day, than from the equipment. And, yes, cigarettes are much more carcinogenic.
} This reply is not for starting a polemic but just to tell that ,we, } manufacturers of microscopes are responsible people and it's also our } health when we are working on theses machines.
I'd say that EM manufacturers are well above average in this respect. Yours, Bill Tivol
"ribbon doesn't have the K transfer. The only dye-sub I'm aware of with includes K with CMY is Tektronix ... I'd like to know of others. "
Silly, commment! The Kodak 8600 and 8650 series both can utilize the CMYK (and K only) ribbons. Afterall this is the engine upon which the N16xx's are built!
Next comment: ...and this is a really scary one. Since we couldn't afford the nice interface of the Codonics NP1600/1660 we just got delivery of a Kodak DS 8650 PS ($7k after current $1500 Kodak rebate). O.k. Last fall in desperation for something to print color we bought an Epson Photo-stylus for $325.00. After printing the exact same image (digital composed at true 800 dpi resoultion) to both the Kodak ($3.50 / page - 300dpi) and the Epson (glossy 'film' media $2.20/page - 720 dpi) a series of comparisons were made, by experience EM users and photographers here with the result being that (1) beyond 12" you can not see a difference other than the fact that the Espon print has a little sepia tint to it, (2) "Up-close / In-your-face" the ink dots are detectable in the Epson prints and there is a slight presence of the 'printer head lines' in the deepest solid black areas (i.e. print something solid black on a laser printer and look for horizontal lines), (3) final difference Kodak's extralife coating has an archive life expectancy of } 50yrs (well its lasted 5 days so far! We'll just have to wait and see.), (4) both printers take approximately the same amount of time to print (O.k., the Condonics is "the fastest on the market", and wasn't run in a side by side comparison).
Now on the cheaper papers (Media) for the Ink jets you get a very nice image quality (albeit not that'll match the dye-sub, but still very acceptible) you start dropping the price/page alot, i.e. $0.75/page, $0.24/page, $0.10/page (...then you hit laser printers, anyway). And for work prints, reviewer copies, etc. these are very good.
Lets see $14,000 printer = $325 Epsons x 43 units ....
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
In reference to your conversation on printers... You are right that the Epson printer has more bang for the bucks in my opinion. I have been selling the dye-subs for some years now. After doing umteen side by side demos you can hardly tell the difference between the two. i let the customer do the comparison... By far the Epson Stylus 800 and not the Epson photo Stylus is better. it does 1440 DPI.. I suggest the Photo glossy paper from Hammermill. Others such has the new paper from polaroid are sort of yellowish to start. That is kind of hard to notice unless you do a hands on comparison. The Epson Stylus 800 going now for about $350 with the paper running about a buck a print... Check it out, You'll be amazed....CP -----Original Message-----
"ribbon doesn't have the K transfer. The only dye-sub I'm aware of with includes K with CMY is Tektronix ... I'd like to know of others. "
Silly, commment! The Kodak 8600 and 8650 series both can utilize the CMYK (and K only) ribbons. Afterall this is the engine upon which the N16xx's are built!
Next comment: ...and this is a really scary one. Since we couldn't afford the nice interface of the Codonics NP1600/1660 we just got delivery of a Kodak DS 8650 PS ($7k after current $1500 Kodak rebate). O.k. Last fall in desperation for something to print color we bought an Epson Photo-stylus for $325.00. After printing the exact same image (digital composed at true 800 dpi resoultion) to both the Kodak ($3.50 / page - 300dpi) and the Epson (glossy 'film' media $2.20/page - 720 dpi) a series of comparisons were made, by experience EM users and photographers here with the result being that (1) beyond 12" you can not see a difference other than the fact that the Espon print has a little sepia tint to it, (2) "Up-close / In-your-face" the ink dots are detectable in the Epson prints and there is a slight presence of the 'printer head lines' in the deepest solid black areas (i.e. print something solid black on a laser printer and look for horizontal lines), (3) final difference Kodak's extralife coating has an archive life expectancy of } 50yrs (well its lasted 5 days so far! We'll just have to wait and see.), (4) both printers take approximately the same amount of time to print (O.k., the Condonics is "the fastest on the market", and wasn't run in a side by side comparison).
Now on the cheaper papers (Media) for the Ink jets you get a very nice image quality (albeit not that'll match the dye-sub, but still very acceptible) you start dropping the price/page alot, i.e. $0.75/page, $0.24/page, $0.10/page (...then you hit laser printers, anyway). And for work prints, reviewer copies, etc. these are very good.
Lets see $14,000 printer = $325 Epsons x 43 units ....
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
Halogen bulbs in general use do produce a significant amount of UV radiation. The halogen lamps sold for home use now all have a simple UV filter to reduce the exposure. UV is readily filtered and as long as you are not looking at the light directly there should not be a problem. If you will be spending a lot of time looking at the light directly, that is not filtered by passing through some other material first, it might be a good idea to place a simple UV fiter in front of the bulb.
**** Keith Ryan wrote: I would not think that you have any real problem. These bulbs are used generally in microscopy, slide projectors, car headlights and probably other uses too. All lamps will produce a spectrum of radiations but not excessively in the UV band unless designed to do so specifically, like high pressure mercury lamps for fluorescence microscopy. Most simple light microscopes don't incorporate UV filtering in their setup.
Common sense says don't spend long looking at it i.e. don't expose your eyes directly to the light! **** Phil Dahlstrom PLDahl-at-aol.com
I have just completed putting the Microscopy & Microanalysis'98 Search Engine on-line at:
http://www.msa.microscopy.com
Just follow the links to the Microscopy & Microanalysis'98 Meeting Page. Using this engine you may search the program by Author, Keyword, Symposium and/or day of the week. The data base is not perfect, so if you find any errors please report them to me off-line.
The Program Committee has once again done an excellent job and it promises to be an great meeting as always. See you there....
My comment on the potential source of X-Rays from the side-arm of our TEM was not a recommendation for people to operate in this manner. For the record our TEM is 10 years old and the tip was given to us by an experienced TEM engineer from the manufacturer. We were testing for radiation and were not getting any counts ( except from the Manager's old watch ! ). He suggested this as a means to confirm that the test equipment was working. I have no idea whether any Jeol TEM's of a similar age would perform in the same way. Perhaps an owner of a JEM 1200 would like to test this ? I do agree that the manufacturers have been extremely safety conscious in the development of modern EM's. Older EM's unfortunately were not as safe. Also I am sorry to be the barer of bad news but some of your colleagues are not in the best of health, since a number of them that I have known personally have passed away over the last number of years. One of my own colleagues died from cancer in her 30's. Electron Microscopy, & associated preparative techniques, can be a hazardous environment. All we can do is be as safety conscious as possible and try and minimise the risks.
Colin Reid
Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
} Hello } } I would like to know what is your EM. } As a field engineer since 22 years, I can tell you NO SEM and NO TEM from } any manufacturer has Xray leakage } The problem is coming and at that point I agree with Mr Garber, when the } people has modified or replacing original parts with local supply for example. } Speaking about the microscope I know, Jeol, It's for example impossible to } generate beam on a 2010 (200kv) with no } sample holder, there is an opto coupler detector. Of course if you insert } something that simulate the holder } you can have the beam. But this is a wrong manipulation. Like to remove a } Gamma source from it's lead castle. } You can do it but you have to think about that because you have be warned! } Same for electron microscopy. } May be you can check the statistics about cancer in the EM environment, for } myself since all that years, all my } collegues and competitors are still living and I'm sure that cigarettes } cause more painfull than EM radiation. } This reply is not for starting a polemic but just to tell that ,we, } manufacturers of microscopes are responsible people and it's also our } health when we are working on theses machines. } Cheers. } } =================================== } Jacky Larnould } JEOL Europe SA Service Manager } mailto:larnould-at-worlnet.fr } mailto:larnould-at-mnet.fr } WEB: {http://www.jeol.com} } Phone:33 (0)4 67 72 28 26 } Fax :33 (0)4 67 79 54 90 }
We have been using an Epson 800 for a few months now and have produced images on Epson Glossy Film which have been accepted by customers in plac= e of conventional photos. Excellent quality but shame about the price (=A31.80/sheet) ! I have just tested some paper from a supplier ( "own = brand paper" ) here. The paper is 140g Glossy at a cost of 7p/sheet ( about 1= 0 cents ). The quality is excellent but it crinkles slightly. Still for the price !
Colin Reid Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
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--Society for Ultrastructural Pathology--
Ultrapath IX will be held in beautiful Asheville, North Carolina, August 2-7, 1998. As in years past, a variety of topics will be covered and participation will be solicited from those attending. Those wishing to contribute to the scientific program are urged to contact Allan Tucker at the address below. Evening case presentations and poster board sessions will be held. A syllabus containing a summary of each presentation will be distributed to all participants. Among others, Dr. Feroze Ghadially of Canada and Dr. Doug Henderson of Australia are now confirmed speakers. The scientific program will emphasize the applications of immunocytochemistry, electron microscopy, in situ hybridization and related techniques in diagnostic pathology. The registration fee is $495.00 and includes the fee for attendance at all lectures and case presentation sessions, CME credit, syllabus, coffee breaks, five breakfasts, two luncheons, opening reception, excursion trip and two dinners.
The course will be held at the beautiful Grove Park Inn Resort in Asheville, NC (http://www.groveparkinn.com). The resort features an 18 hole championship golf course, as well as six outdoor and three indoor tennis courts, an indoor sports center, and indoor and outdoor swimming pools. The resort is located in the Blue Ridge Mountains of western North Carolina. Hiking, fishing, white water rafting, horse-back or mountain bike riding and other activities are available nearby. Daily jet service via the Asheville Regional Airport is available. A variety of recreational activities are planned for registrants and their families.
For further information, please contact either:
J. Allan Tucker, M.D. Department of Pathology University of South Alabama 2451 Fillingim Street Mobile Al 33617 Tel:334-471-7473 or 334-471-7799 Fax: 334-471-7884 or 334-660-5576 email: JTucker-at-usamail.usouthal.edu
or:
John Shelburne, M.D., PhD Department of Pathology Box 3712 Duke University and VA Medical Centers Durham NC 27710 Tel: 919-286-6979 or 919-286-6925 Fax: 919-286-6818 or 919-684-8689 email: shelburne.john-at-forum.va.gov
URL for Online Information: http://sup.ultrakohl.com/ultrapat.htm
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Opinions aside (and there are many!), the only true test of printer comparisons is for world-wide colleagues to take a digital image and find a vendor which can generate sample prints from the various print technologies available: inkjet, laserjet, dye-sublimation and hybrid. Also, relating the printer technology to what is applied in the darkroom will yield an understanding of what constitutes an acceptable image. Personally I have done the aforementioned with a series of digitally-captured TEM images. Basically, for our personal needs, TEM negatives have been printed using a high-contrast point light source. Diffuse-light sources generate softer edges (less contrast) which for us has been acceptable for black & white LM histology images but not for TEM. The technology of dye-sub printers (subliming the dye to a gaseous state which permeates the fibers of the print media) results in quality similar to using a diffuse-light source with TEM negatives--everything has a soft edge. Inkjets are high-contrast technologies and the high-resolution Epson (1440 dpi on photo-quality glossy paper) results in truly remarkable, near-photographic images (held at some distance away. Close examination reveals pixelation). The Epson also more closely matched in color balance a true TEM photographic print (print options set to BLACK ONLY--not COLOR). Finally, the best bang for the buck is outputted from the Fujix Pictrography, a hybrid type of technology that uses silver halide for image formation--so it acts like a Polaroid for image processing, but the results are darkroom photography! However, due to cost factors, we seem to be headed in the direction to print TEM image "thumbnails" on our dye-sub (Black Ribbon ONLY-not color) and generating manuscript prints on the Pictrography (both TEM and color LM). There--now I can get down off the Dye-Sub soapbox!
Best regards,
Walt Bobrowski Subcellular Pathology Parke-Davis Research Ann Arbor, MI 48105
The cheap FARGO PrimeraPro thermosub printer also prints with CMY, CMY+Overlay (same ribbon) OR with a CMYK ribbon OR with a K ribbon (b/w). Resolution 300x600 max. We bought one, it works well, but I don't have possibility to compare it with others, so can't state much about that. We bought it because of tests in computer journals (don't remember which) and our limited money.
wrote:
} But if I were to wish for perfection, it would be that it printed with } a CMYK ribbon rather than to create grayscale images with CMY. I don't } know that any dye-sub will print grayscale without a tint if their } ribbon doesn't have the K transfer. The only dye-sub I'm aware of with } includes K with CMY is Tektronix ... I'd like to know of others. (... } BTW ... All dye-subs will offer a K ribbon ... but switching between } color and grayscale printing is not practical ...)
Does anyone have experience with the Alps dry-ink printers? I'm interested in the Epson 800 or 1520 printers to take the burden off of our Tek Phaser IIsdx. They are listed at 720 dpi, yet with less bleed they might rival higher dpi inkjets, and the ink is water resistant.
TIA, -- Glen MacDonald Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax The box said "Requires Windows95 or better". So I bought a Macintosh.
Dear all, Please excuse the slightly non-microscopy topic of this posting.
I have an Epson Stylus Color 800 inkjet printer (with which I am very happy). The cost of Epson's own brand A4 OHP transparency slides is pretty high, though (} =A31 each). Are there aternative brands that work just as well but cost less or am I best of sticking to the manufacturers own brand?
Thanks for any advice, opinions etc. that you can give.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
It was not my original intention to post a summery but I have received more request for the summery than I have responses from users. The # of responses was small ( {10) but then I only asked for experiences on systems less than a year old.
Owners & users of video systems were generally happy with their systems. People with slow scan cameras that were in operational order were also generally content with the system performance. The catch here is "operational order" By far the major complaint in dealing with Gatan is that of timely delivery & getting online when promised. One respondent stated that delays were so great that they nearly lost their funding prior to delivery. Apparently field service leaves much to be desired. Salesman failing to return voice & e-mail was complaint. Salesman also seemed to be oblivious to the physical constraints of mounting their hardware in our confined spaces. I guess just like in a lot of other industries salesman are the problem, the 1st source of disinformation & false expectation.
A few comments on specific pieces: Model 789, a side mt. video camera "quality of the images is barely adequate" "not publication quality" " used primarily for demonstrations"
Model 673/III , video camera "expensive but extremely useful " "sensitive ...used for low-dose cryo" "low res & less than 8 bit gray scale, effectively"
Model 791, MultiScan camera " happy with hardware & software" " a very polished package"
MultiScan camera, model unknown. Gatan can't get software configured on G3 MAC, tech left & has yet to return.
Locally, Gatan has failed miserably in the tech support & software compatibility areas. I was a bit surprised at this report but knowing the source I considered it valid. My primary purpose for surveying the user group is to find out if this is a local or global problem. It seems global. Based primarily on this perception we have elected not to pursue a Gatan Multi-scan camera this year. Perhaps when Gatan is in better light we will reconsider, after all this type of system is a part of the modern hi-res TEM. In general people are happy with Gatan once installed & operational but the transition from salesman blowing air to operational seems riddled with delays. No one is impressed by the cost of the systems or upgrades. Again once operational there eq. performs as expected. For some applications Gatan is the way to go. Retrospectively I am sure most respondents will look closer at competitors next time around. For a lot of applications one can throw down 5-10K$ for a good digital negative scanner. Then there is a broad choice in software.
One thing I had hoped for but got no feed back on is Gatan's PC based software. I thought it had been released., maybe not. Can anyone address this subject?
Cell Vision Subscription Price List 1998 (Now included in Index Medicus and Medline)
Cell Vision publishes peer-reviewed scientific, medical and technical = articles in the fields of pathology, morphology, anatomy, histology, = microscopic sciences, etc., 6 times per year.
For individuals: $130 per year For institutions: $150 per year Shipping and handling: within USA add $6, international add $20.
Special price as a member of the International Society of Molecular = Morphology (Oklahoma City, OK, USA; information: Phone: (609) 802-0242, fax: (609) = 802-0245, e-mail: ismmcvjg-at-uscom.com)
Please make check payable to: Cell Vision, Institute of Molecular Morphology, 560 Fellowship Road, = Suite 407, Mount Laurel, New Jersey 08054, USA Phone: (609) 802-0242, fax: (609) 802-0245, e-mail: ismmcvjg-at-uscom.com
CELL VISION: NEW INTERNET WEB-SITE:
http://www.cbba.org/cbba/Default.htm Still under construction
Thank you for your interest. Yours respectfully
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
DISCLAIMER
Disclaimer: The views expressed in this e-mail message do not necessarily represent = the official views of the Inst. Anatom. Pathology LKA, from which this = message was conveyed. No commercial interest in products/product lines, company/-ies, = societies, if such names are mentioned or such are refered to. No = liability for incidentally incorrect information, no warranty: I'm = sharing only my daily experience.
I have some users who want to make high resolution scans of 4x5 TEM negatives. The 45 series scanners from Nikon, Polaroid, et al run US$8,000-9,000. However, I've been told that they are barely capable of 3.0 OD at 4"x5". The Imacon flatbed will deliver 3.9 at 4"x5", but it is about US$16,000. What are others using for scanning negatives?
TIA, -- Glen MacDonald Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax The box said "Requires Windows95 or better". So I bought a Macintosh.
We have an Alps printer and are very happy with it. We have it for the same reason you would like to have one-- to remove some of the pressure on our Tek Phaser IIsdx.
Patty Jansma Tel:520-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
On Mon, 20 Apr 1998, Glen MacDonald wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anyone have experience with the Alps dry-ink printers? I'm } interested in the Epson 800 or 1520 printers to take the burden off of } our Tek Phaser IIsdx. They are listed at 720 dpi, yet with less bleed } they might rival higher dpi inkjets, and the ink is water resistant. } } TIA, } -- Glen MacDonald } Virginia Merrill Bloedel Hearing Research Center } Box 357923 } University of Washington } Seattle, WA 98195-7923 } glenmac-at-u.washington.edu } (206) 616-4156 } (206) 616-1828 fax } The box said "Requires Windows95 or better". So I bought a Macintosh. }
I have an AMRAY 1830 SEM (with joystick for stage control) and EDAX 9900. Now I am thinking about getting a PC interface for the SEM and EDX using my very, very limited funding available. The features must include SEM image acquisition and storage(SEM control and image processing are not needed), stage control (in X,Y), unattended EDX acquisition and X-ray mapping(with Be window open/close control if possible). Anybody has such experience and would suggest a solution? Any response is appreciated.
Tong
-------------------------------------------------------------- Tong Wang email: tong-at-jlab.org MS 58, Jefferson Lab URL: http://www.ee.odu.edu/~xxu 12000 Jefferson Avenue fax: (757)269-7658 Newport News, VA23606 tel: (757)269-7334 --------------------------------------------------------------
I believe that TEM film has a density range of about 2.8-2.9. If this is true, any scanner with a range of over 3 should work.
Henk
} } I have some users who want to make high resolution scans of 4x5 TEM } negatives. The 45 series scanners from Nikon, Polaroid, et al run } US$8,000-9,000. However, I've been told that they are barely capable of } 3.0 OD at 4"x5". The Imacon flatbed will deliver 3.9 at 4"x5", but it } is about US$16,000. } What are others using for scanning negatives?
} TIA, } -- Glen MacDonald } Virginia Merrill Bloedel Hearing Research Center } Box 357923 } University of Washington } Seattle, WA 98195-7923 } glenmac-at-u.washington.edu } (206) 616-4156 } (206) 616-1828 fax } The box said "Requires Windows95 or better". So I bought a Macintosh. } } Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
We use flatbed scanners from either Perkin-Elmer or Zeiss. Both, however, are fairly expensive, say well over 50 k$ :-(. But, they deliver for that kind of money (of corse they'd have to ...). What kind of scans are the users interested in? And, on the OD, most scanners that I know of begin to run of headroom at 3 OD. J.
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Mon, 20 Apr 1998, Glen MacDonald wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have some users who want to make high resolution scans of 4x5 TEM } negatives. The 45 series scanners from Nikon, Polaroid, et al run } US$8,000-9,000. However, I've been told that they are barely capable of } 3.0 OD at 4"x5". The Imacon flatbed will deliver 3.9 at 4"x5", but it } is about US$16,000. } What are others using for scanning negatives? } } TIA, } -- Glen MacDonald } Virginia Merrill Bloedel Hearing Research Center } Box 357923 } University of Washington } Seattle, WA 98195-7923 } glenmac-at-u.washington.edu } (206) 616-4156 } (206) 616-1828 fax } The box said "Requires Windows95 or better". So I bought a Macintosh. }
About a month ago I posted a request for information regarding resellers = of used optical microscopes. Since then, I've gotten a number of = requests to summarize the information I received. So, without further = ado:
Websites:
LabX www.labx.com Labequip www.labequip.com Montana Microscope www.montanamicroscope.com Conneaut Lake Scientific www.wwweb-pro.com/cls Used Mechanical Equipment www.execpc.com/ume Lehman Scientific www.lehmanscientific.com Capovani Brothers www.capovani.com Scientific Equipment Exchange www.sci-equip-ex.com Bid Service www.bid-service.com
Plus, a web search of "used microscopes" using your favorite search = engine will turn up a few others.
Phone Contacts:
Martin Microscope Co (864) 242-3424 or (864) 859-2688 Bob Martin Mel Sobel 1-888-ALL-SCOPES or (516) 935-7794 Technical Instruments (415) 431-8231 Rick Staples=20 Bay Optical (415) 431-8711 Tom Henry ARC Instruments (606) 498-1345 Phil Hutcheson
There's a ton of junk equipment out there, but after searching long and = hard, we were able to come up with what we were looking for.
Many thanks again to all who responded to my original post for = information!!!
Tom
Thomas C. Isabell, Ph.D. Research Scientist E.A. Fischione Instruments, Inc. tc_isabell-at-fischione.com webpage: www.fischione.com
I have to buy a new vacuum gauge, (heads and controller), and my short list is either Leybold or Balzers (having eliminated Edwards because of their frequent model changes, which lead to "sorry, no service now available for that model" after not many years). Oh for the good old analogue-and-repairable-for-ever days! I have no experience with either brand, anyone got any good or bad news about either?
Confidentiality guaranteed for personal replies.
thanks
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
I haven't timed it for a file that size. An A4 page with two of my imag= es would be approx. 2MB and would print in about 2-3 mins at top resolution. Routinely I print the greyscale images on a 600dpi laser and only use the Epson for higher quality. The configuration of your computer will help = to a certain extent. I am lucky in that it is attached to my spare 200MHZ = PC so I don't have to sit and watch it.
Colin Reid Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
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I have to buy a new vacuum gauge, (heads and controller), and my short list is either Leybold or Balzers (having eliminated Edwards because of their frequent model changes, which lead to "sorry, no service now available for that model" after not many years). Oh for the good old analogue-and-repairable-for-ever days! I have no experience with either brand, anyone got any good or bad news about either?
Confidentiality guaranteed for personal replies.
thanks
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
we decided on a different set-up: we purchased a MicroLumina camera with a high frequency lightbox. We wanted to scan not only TEM negatives but also X-ray films and a series of old 35 mm slides. Besides we are using the set-up for macro-photography as well. The camera is mounted on a photographic stand, which allows one to adapt the camera-film distance always to have the full size negative/object in view and therefore the full camera resolution on the object. The camera resolution is 3000x2600 pixels and sells for less the US$ 9k. Again, as the camera is adjusted to the object size, this resolution is available for a TEM negative as well as a 35 mm slide. The only drawback is each scan takes about 1-2 minutes as the camera is based on a line array.
Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
} ---------- } From: Glen MacDonald[SMTP:glenmac-at-u.washington.edu] } Sent: Monday, April 20, 1998 5:12 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Negative Scanners } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We use all three, Codonics, Epson Stylus, and Fargo Primera Pro printer. We are most happy with these printers in the order listed. Quality of print is very important to us, but time is, too. The two latter printers have taken upwards of 30 minutes to print many of our requested TEM and Confocal images. The Codonics never takes that long for tif, bmp, or postcript images. The Epson and Fargo both require that you have specialized software like Adobe,for example and drivers that will make their printer work. They print only a limited image type. The Codonics has all the conversion hardware/software built in and can be placed any where on the network ! A tremendous feature for us. Since the entire University can send prints to it, and they can use almost any format, bmp, tif, gif, eps, ...ETC. with almost no further need to adjust their image. Since we support 1000's of prints every year, and sometimes 1000 monthly, near grant writing time, we find the Codonics indispensable. We find the 300 pixel per inch resolution image is very similar to the 1200 dot per inch image you get off other printers, but there is no pixelation when looking closely at the image (if you started with a good image). The reproducibility is astounding. The first of 20 images looks just like the last one. And it takes about 20 minutes to do 20 images. None of the others would do that for us. For example, although the Epson comes close, it would takes us about 6 hours or more to do 20 images at high resolution, so we did not even try to do that. The other problem, is that there does not seem to be an easy way to send an image and get 20 prints from the Epson or Fargo printers, a software problem I guess. We can not have someone stand by and send a print every 30 minutes. Small things, but that costs us time. We now use the Epson for fast-low-res images (at 300 dpi) when needed since it has that capability and we can use plain paper. But the low cost per print is still about 35 cents. The Codonics, price per print has recently gone down for us, bulk buying, to about $2.00 print. We are hoping to see improvements on this possibility. After an extensive survey about two years ago and after multiple copies (over 800) we were overwhelming advised by research faculty (99%) to purchase the Codonics. We are very happy with it, and the service record is also outstanding. They have always found a way to get us support QUICKLY whenever we needed it. Supplies are readily available and are expected to be for a long time. KODAK is the major supplier for their machines and for Codonics with the Kodak engine. However, Codonics, does re-package the ribbon and paper and supports their product 100%. Feel free to contact me for a more detailed discussion if you wish. Technical Coordinator for Microscopy, Tom Baginski, 301 295 5691 or email:tombg-at-bictom.usuf1.usuhs.mil DOD/USUHS/BIC, Bethesda, MD
We have an ALPS MD-2300 which does dye sub and microdry printing. The dye sub technique produces images good enough for publication but is quite slow, requiring nearly 3/4 hr for an 8"x10" print. The standard microdry printing is fine for rapid display of CMYK images if one uses their special paper. The printer is finicky about other, less expensive paper, but can be gotten to work. I find their monochrome black non-dithered half toning a little annoying even at 600 dpi, but usable. Hope this helps.
Len } Does anyone have experience with the Alps dry-ink printers? I'm } interested in the Epson 800 or 1520 printers to take the burden off of } our Tek Phaser IIsdx. They are listed at 720 dpi, yet with less bleed } they might rival higher dpi inkjets, and the ink is water resistant. } } TIA, } -- Glen MacDonald } Virginia Merrill Bloedel Hearing Research Center } Box 357923 } University of Washington } Seattle, WA 98195-7923 } glenmac-at-u.washington.edu } (206) 616-4156 } (206) 616-1828 fax } The box said "Requires Windows95 or better". So I bought a Macintosh. } } Leonard Zablow Howard Hughes Medical Institute 722 West 168 St. New York, N.Y. 10032
Jon, A few years ago I ordered a Beseler negative carrier #8343, 4x5 anti-newton from Brandons, Inc. in Jacksonville, FL (800-874-5273). It has glass for supporting the film. I use it for my TEM negatives. I think they special ordered it for me. I would also suggest searching the web for Beseler and try to buy it directly from them.
best regards, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
We use all three, Codonics, Epson Stylus, and Fargo Primera Pro printer. We are most happy with these printers in the order listed. Quality of print is very important to us, but time is, too. The two latter printers have taken upwards of 30 minutes to print many of our requested TEM and Confocal images. The Codonics never takes that long for tif, bmp, or postcript images. The Epson and Fargo both require that you have specialized software like Adobe,for example and drivers that will make their printer work. They print only a limited image type. The Codonics has all the conversion hardware/software built in and can be placed any where on the network ! A tremendous feature for us. Since the entire University can send prints to it, and they can use almost any format, bmp, tif, gif, eps, ...ETC. with almost no further need to adjust their image. Since we support 1000's of prints every year, and sometimes 1000 monthly, near grant writing time, we find the Codonics indispensable. We find the 300 pixel per inch resolution image is very similar to the 1200 dot per inch image you get off other printers, but there is no pixelation when looking closely at the image (if you started with a good image). The reproducibility is astounding. The first of 20 images looks just like the last one. And it takes about 20 minutes to do 20 images. None of the others would do that for us. For example, although the Epson comes close, it would takes us about 6 hours or more to do 20 images at high resolution, so we did not even try to do that. The other problem, is that there does not seem to be an easy way to send an image and get 20 prints from the Epson or Fargo printers, a software problem I guess. We can not have someone stand by and send a print every 30 minutes. Small things, but that costs us time. We now use the Epson for fast-low-res images (at 300 dpi) when needed since it has that capability and we can use plain paper. But the low cost per print is still about 35 cents. The Codonics, price per print has recently gone down for us, bulk buying, to about $2.00 print. We are hoping to see improvements on this possibility. After an extensive survey about two years ago and after multiple copies (over 800) we were overwhelming advised by research faculty (99%) to purchase the Codonics. We are very happy with it, and the service record is also outstanding. They have always found a way to get us support QUICKLY whenever we needed it. Supplies are readily available and are expected to be for a long time. KODAK is the major supplier for their machines and for Codonics with the Kodak engine. However, Codonics, does re-package the ribbon and paper and supports their product 100%. Feel free to contact me for a more detailed discussion if you wish. Technical Coordinator for Microscopy, Tom Baginski, 301 295 5691 or email:tombg-at-bictom.usuf1.usuhs.mil DOD/USUHS/BIC, Bethesda, MD
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Listserve members, I will be traveling to P.R. of China in a few weeks and will, with my husband, be visiting Biology Departments at a university in Beijing and a university in Heifei where I will be meeting with microscopists. I would like to bring along a few inexpensive items which may be difficult to get in China for our hosts. One friend has already asked for double-sided scotch tape. Any suggestions for other small items useful in microscopy labs would be appreciated. Please reply directly to me as I don't know if this would be of general interest to the whole list.
Debby Sherman, manager Phone: 765-494-6666 Microscopy Center in Agriculture Fax: 765-494-5896 Purdue University E-mail: sherman-at-aux.btny.purdue.edu W. Lafayette, IN 47907 or: emcenter-at-btny.purdue.edu
by huey.cadvision.com (8.8.5/8.8.5/DCE/TRI) with ESMTP id HAA55576 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 21 Apr 1998 07:36:10 -0600 Message-Id: {199804211336.HAA55576-at-huey.cadvision.com}
We currently use an older Leaf 45 film scanner for film format from 35mm to 4 x 5. These scanners are often available from digital image equipment supply houses such as B & H in New York (212) 444-5008 0r Graphtronix (612) 461-5151. Although I have never had dealings with these companies myself, they seem to have good reputations in the digital imaging communities. A 4 x 5 scan on our leaf produces a file size of ~82Mb. The dpi is 1200 and the image is 6000 x 4740 pixels. These are for a landscape orientation. For portrait, the figures are 1200 dpi, 4760 x 4740 pixels and a file size of ~67Mb. The scanner accommodates a standard Beseler 4 x 5 negative carrier. If anyone in the research community is interested, we will be glad to scan a few negatives for experimental purposes. Just send the neg. along with either a Zip or Jazz disk and we will send back some scanned files. Since we do this for a commercial venture, please only academics or research inquiries. If you want a commercial price please contact us off of this list server at our e-mail.
Jon McGovern J. P. McGovern and Associates e-mail: jcgover-at-cadvision.com (403)291-3196 (403)291-1423 Fax
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Hi there
I have to buy a new vacuum gauge, (heads and controller), and my short list is either Leybold or Balzers (having eliminated Edwards because of their frequent model changes, which lead to "sorry, no service now available for that model" after not many years). Oh for the good old analogue-and-repairable-for-ever days! I have no experience with either brand, anyone got any good or bad news about either?
Confidentiality guaranteed for personal replies.
thanks
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
by mail.unixg.ubc.ca with smtp (Exim 1.71 #1) id 0yRgHA-0000SX-00; Tue, 21 Apr 1998 09:52:08 -0700 Message-Id: {2.2.32.19980421165018.008df1a0-at-pop.interchange.ubc.ca} X-Sender: mager-at-pop.interchange.ubc.ca X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Dear Jonathan, There is a special negative holder for Polaroid 4" X 5" negatives, made by Beseler. We use it for the TEM negatives, turned sideways. When the original broke, we purchased a new one from our local photography store. The other one we have looks homemade, before my time, which has a hinged holder consisting of two metal plates that fits the Beseler, with two glass plates held in cutouts. The negative is sandwiched between the glass plates. It holds up to 4" X 5". The disadvantages is that you must make sure your negative and 4 sides of glass are clean. You wrote: } Hi: } } We inherited a Beseler 45M enlarger, but no negative holder for 3 1/4" x 4" } TEM film. } } We use a crude, homemade holder now. A real one would be nice. Anyone have } an extra or know where I can get one? } } Thanks } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
I am looking for information to procure proceedings from the Pfefferkorn Conferences. The 1983 proceedings was published by Scanning Electron Microscopy, Inc. Does anyone have information on these proceedings?
I am interested in comparing volcanic ash from different sources. If you have ash samples that you would be willing to part with, please email me directly. Thank you!
Bob Willis ManTech Environmental Technology, Inc. willis.robert-at-epamail.epa.gov
you will find there representatives (addresses and phone/e-mailnumbers)
Another www-page I could offer to you is:
http://www.pfeiffer-vacuum.de=20
(German branch of PFEIFFER VACUUM TECHNOLOGIES, perhaps or certainly you = will find on their web-page connections to representatives in your area)
hope this helps, best of luck
Wolfgang MUSS
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: Ritchie Sims[SMTP:r.sims-at-auckland.ac.nz] Gesendet: Mittwoch, 22. April 1998 11:53 An: microscopy-at-sparc5.microscopy.com Betreff: Re: Vacuum Gauges TEM Hardware: vacuum SEM/TEM
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Hello
Does anyone from Edwards High Vacuum USA read this list? If so, please=20 contact me. Alternatively, does anyone have an email address for them?
thanks
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 =20 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 =20 Auckland New Zealand
I have a Leitz Laborlux D microscope at home and I'm trying to find a 'C' mount and a 2.5 or 3.3X photoeyepiece for it. If anyone has thes and is willing to part with them I'd like to buy them from you. Yes, I'm aware that I could probably get them from Leica but dollers (mine) are a concern.
TIA
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
This response is to an inquiry concerning a particular high resolution sputter coater and a reply by its first user, Mel Dickson. That posting is a few days old but requires a response and I sought and have included here comment from EMITECH (UK). There is nothing wrong with Mel's comments, except that without further discussion the topic is treated one-sidedly. Clearly, the Xenosput works well, but its suitable for very small samples only; its expensive to purchase and the Xenon gas which is heavier than argon and hence more efficient, is also rather expensive to buy. There is an amazing range of coating equipment on the market, all with their particular pros and cons.
"High resolution SEM has created a requirement for high resolution coating. Typically chromium is now used as the target material, primarily because chromium has a much finer grain size than gold. It also does not readily form coagulated islands, as is the case with gold and that is the limiting factor of conventional gold sputter coatings in high resolution SEM.
However, for Cr sputtering the vacuum system must be kept clean and the energy levels for sputtering increased to allow sputter cleaning of the target prior to sputtering. This has resulted in a new generation of sputter coaters like our Emitech K Series, which includes the K575 and the K675. The K575 can readily coat evenly an area of 76mm diameter, and the K675 which was designed specifically for the semi conductor market's 203mm wafers, can coat up to 253mm diameter. Such instruments are turbo pumped and fully automatic and employ a flushing and cleaning protocol to ensure a clean vacuum. They may also have a shutter assembly to protect the specimen during the sputter clean operation. This results in very good quality and repeatable deposition. As well as chromium targets other materials such as platinum should be available. Other desirable features of such instruments are a fully automatic control, with argon as process gas and nitrogen as purge gas. David Robinson, EMITECH Ltd"
Please note that ProSciTech has an interest in EMITECH equipment, since we distribute this equipment, however, we are only a regional distributor for this company (Australia, NZ, parts of SE Asia). Considering our profile in the home market, I don't see any commercial advantage from posting this item here. I just like to contribute to this topic which deserves fuller treatment. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** -----Original Message-----
The address is correct. Thanks for adding me to the list.
Hello: We are using MagOp Disks on Power Macs to archive images in our lab, and I am interested in knowing if anyone has found a Disk Repair Program which repairs or retrieves files from the occasional MagOp disk which starts to have problems.
I need information to educate the administration on how close to "cost neutrality" a microscopy facility can get. I currently supervise a facility with a part-time employee to assist in repairs. Much of the equipment is 20-25 years old and is constantly breaking down. I am the sole revenue generater with a few students who have been trained to use the microscope and have beamtime hours. Within a year or so of operation the facility went from generating $0.00 in revenue to ~$60K maybe 70K/year. This doesn't pay all the bills of an EM Facility with 2 TEMs and an SEM as well. My question is, is their any facility that is COMPLETELY cost neutral with revenue only for service work provided? Are the numbers I describe for a medium sized University reasonable? I would greatly appreciate any input. If their is a University that spends only what they bring in for doing samples in a multi-user environment please contact me I be happy to know how you did this.
Best Regards,
Kirk J. Czymmek, Ph.D. Biological Electron Microscopy Facility University of Delaware Newark, DE 19716 kirk-at-udel.edu
We would be interested in suggestions and collective experience with measuring the energy spread of a FEG-TEM by a method other than using a PEELS spectrometer, i.e. by some type of imaging/diffraction analysis technique. We have been using the standard method of measuring the contrast-transfer envelope and associated information limit by diffractogram analysis of amorphous Ge. However, after some work the accuracy of this method for resolving differences in energy spread even as large as 100% is not clear to us. Assessing the limits of the envelope function by seeing how well the microscope passes various fringe spacing in the 1.2 to 0.9 A range (i.e. out on the limits of the envelope function) seems reasonable, but what are the pitfalls? Other methods using beam tilt effects on diffractograms exist in the literature but remain to be checked out.
Input from you FEG-TEM artists out there would be appreciated.
Roy Christoffersen Materials Science Research and Engineering Center University of Houston 3201 Cullen Houston, TX 77204-5932 roy-at-bayou.uh.edu (713) 743-8273 FAX: (713) 743-2787
We have had good sucess from 1. Taking an EDTA tube of blood and spin it down 2. Take a long nosed pipet and collect the buffy coat area, roughly. 3. Take a Wintrobe tube ( tall skinny tube used in hematology) , place rough buffy coat in this tube) 4. Spin down the wintrobe tube. 5. Remove supernant, then collect Buffy coat with a thin glass pipet. 6. Layer gently in bottom of a vial of Karnovsky's and let set 20-30 min.
* At this point it should look like a fat miniture snake at the bottom of the vial.
7. Process gently through the rest of the EM procedure. __ should the buffy coat break up, place in ependorf tube and gently spin before each change in a microcentrifuge tube, low speed.
8. If the buffy coat is intact, by embedding time, cut with razor and seperate among embedding mold/capsules.
___ If the buffy coat is not intact, place in very small beam capsules, and put as concntrated of drops of samples as possible into the beam capsules.
__ Fill, close, and place in 1.5 ml ependorfs and spin in the microcentrifuge for 15-20 minutes at moderate speed. Polymerize as is in the capsule/ependorf assembly. At some point after fairly hard, razor off the eppendorf for better polymerization.
Hope this helps, just one way to try,
Lou Ann
Winnie Wrote: Hi There, Looking for a good technique for making a pellet of buffy coat from normal blood. Would appreciate any help. Thanks.
Winnie
***************************************-at-redfoot Lou Ann Miller Center for Microscopy & Imaging College of Veterinary Medicine Dept. of Veterinary Biosciences University of Illinois Rm 1108 Basic Sciences Bld 2001 S Lincoln Ave. Urbana, Illinois 61802
We are interested in buying a single- or double-tilt heating stage that = will fit a JEOL 2000 side-entry goniometer. I believe this goniometer is also = shared with the 1200, 200 and perhaps even their 100 series microscopes. I'm = not sure about the 4000. We would require an attainable temperature of 600=B0C. = At this time we are looking at buying a used stage, and probably won't have the = money to buy new. If you have a stage that fits this description and would = consider selling it, please contact me at the address below (email, snail mail, = phone or fax). =20
Thanks,
JSV *************************** John S. Vetrano Sr. Research Scientist Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
Does anyone know starting recipes for using cold water fish (teleost) gelatin as a blocking agent for immunocytochemistry? Does it work? Any pitfalls? We have a particularly sticky primary with lots of small punctate background appearing..controls with secondary alone are clean...presently using 0.5% BSA with 1% goat serum as block 1hr r.t....its not stopping it????
I am looking for software that can model the energy deposited by the electron beam in a thin film or bulk sample. Do any of the Monte Carlo codes do this?
I suspect that your punctate signal, when it is background indeed, may come from aggregated primary antibody molecules. In that case using more or different additives to the blocking buffer or incubation buffer will not be very helpful in improving the signal-to-noise ratio. Spinning down the antibody solution (stock, but preferably the diluted antibody) at high speed will help to get rid of aggregates. I think this is the easiest thing to try to begin with. Another explanation might be that there are indeed sticky sites in your specimen. But then one would expect different antibodies from the same animal source to give similar results. I don't know whether that is the case. If so, then fine tuning the blocking and incubation buffer composition is the way to a solution. You may find more info on our web site regarding background at http://www.aurion.nl, and in a previous series of messages to this Listserver sent in the first week of April.
Good luck,
Jan
============================= Jan Leunissen, Ph.D. AURION ImmunoGold Reagents & Accessories Managing Director Costerweg 5, 6702 AA Wageningen The Netherlands
We spin the anti-coagulated blood in a narrow test tube, then carefully remove as much as possible of the plasma without disturbing the buffy coat. Buffered 2% Glutaraldehyde is then very gently layered on top and the tube left to stand in the fridge for about a couple of hours. This gives a buffy coat which is embedded in solid plasma and can be removed from the tube with the help of a thin wooden stick or similar. The resultant disc can then be trimmed and the pieces processed to resin as you would for normal tissue. If you embed in flat moulds your specimen will have orientated layers of plasma, platelets, white cells and red cells.
It's a simple and effective technique, with only one centrigugation to give you a sample which is easily handled. The only downside is that some drugs (e.g. aspirin, I believe) may inhibit the action of glutaraldehyde on the plasma and prevent the conversion of the plasma to a solid.
James Ito Pathology Department Royal Hospital for Sick Children Yorkhill Glasgow Scotland. ---------- } From: Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: COLLECTING A BUFFY COAT FOR TEM } Date: 22 April 1998 21:50 } } Hi There, } Looking for a good technique for making a pellet of buffy coat from } normal blood. Would appreciate any help. Thanks. } } Winnie }
Its easy - if you have an administrator who does not just care about balancing the budget and finds the EM Manager easier to push about than some professor who presides over a poorly performing department. Your EM Unit costs the university real money and ought to be shut down if you supply nothing much in return. I trust that the credit side of your ledger shows that you have sufficient and happy users. In a university they would be mostly grad and some undergrad students and they require many services within an EM facility. Since the university collects money for these students, the unit should get notional or real credit. The "user pays" arguement is poor economics when the script dictates: Collect all the students funding and distribute this to administration, library and departments but give no credits to the electron microscope unit. Too often the EM user has already paid but nothing has gone to the EM Unit and the administrator is in fact looking for a second payment from the user. Kirk, our correspondent's problem is that few administrators are interested in good governance. Most just want to balance the budget along the path of least resistance. Been there, done that, I commiserate. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** } Dear Collegues, } I need information to educate the administration on how close to "cost } neutrality" a microscopy facility can get. I currently supervise a } facility with a part-time employee to assist in repairs. Much of the } equipment is 20-25 years old and is constantly breaking down. I am the } sole revenue generater with a few students who have been trained to use } the microscope and have beamtime hours. Within a year or so of operation } the facility went from generating $0.00 in revenue to ~$60K maybe } 70K/year. This doesn't pay all the bills of an EM Facility with 2 TEMs and } an SEM as well. My question is, is their any facility that is COMPLETELY } cost neutral with revenue only for service work provided? Are the numbers } I describe for a medium sized University reasonable? I would greatly } appreciate any input. If their is a University that spends only what they } bring in for doing samples in a multi-user environment please contact me I } be happy to know how you did this. } } Best Regards, } } Kirk J. Czymmek, Ph.D. } Biological Electron Microscopy Facility } University of Delaware } Newark, DE 19716 } kirk-at-udel.edu } } }
Does anybody have a spare puck (or 2) for a Complot Series 7000 Digitizer (Houston Instruments). After two buyouts of the original company the current manufacturer is not able to help out with this 10 year old (at least) tablet.
Alternately, we are in need of a replacement for the 24" x 36" digitising tablet we were using for measuring area and perimeter (via DOS software package) from projected histological images. We have tried to replace this system with a videomicroscope and image analysis package but cannot get a system to work as well, or as cheaply. Does anybody have a recommendation for tablets of this size, or perhaps 18x24, and some simple software (PC) to get the data we need? Our image analysis software (Optimas) will probably do it, but is a bit of overkill and I figure there must be something simple out there somewhere.
Thanks in advance --------------------------------------------- Rod Dilley Baker Medical Research Institute PO Box 348, Prahran 3181, Victoria, AUSTRALIA fax: +613 9521 1362 email: rod.dilley-at-baker.edu.au ---------------------------------------------
It works well, alone or in conjunction with serum, depending on the application and the nature of the secondaries. Start with a 2% (total protein) solution.
The only drawback is the thick nature of the stock gel. You will have to weight it.
good luck
Ramin Rahbari Tel. 734-622-3383
} -----Original Message----- } From: loudy.d-at-pg.com [SMTP:loudy.d-at-pg.com] } Sent: Thursday, April 23, 1998 12:25 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: cold water fish gelatin } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Does anyone know starting recipes for using cold water fish (teleost) } gelatin } as a blocking agent for immunocytochemistry? Does it work? Any } pitfalls? } We have a particularly sticky primary with lots of small punctate } background } appearing..controls with secondary alone are clean...presently using } 0.5% BSA } with 1% goat serum as block 1hr r.t....its not stopping it???? }
To all who have thus far replied and requested feedback regarding my query about microscopy facility revenue. I can't afford to FAX all the copies that I have received at this point but will mail them to all who have asked once they are compiled.
I will also provide a synopsis to submit to the listserver when I have had the chance to go through all the responses.
Incidently, all costs associated with the facility (salaries, repairs, supplies, incidentals) must be recovered via users fees according to the decree. Is salaries alone were covered, I could actually have enough money to repair or replace some of my more unreliable equipment.
Best Regards,
Kirk J. Czymmek Biological Electron Microscopy Facility University of Delaware Newark, DE 19716 kirk-at-udel.edu
} } Dear Collegues, } } I need information to educate the administration on how close to "cost } neutrality" a microscopy facility can get. I currently supervise a } facility with a part-time employee to assist in repairs. Much of the } equipment is 20-25 years old and is constantly breaking down. I am the } sole revenue generater with a few students who have been trained to use } the microscope and have beamtime hours. Within a year or so of operation } the facility went from generating $0.00 in revenue to ~$60K maybe } 70K/year. This doesn't pay all the bills of an EM Facility with 2 TEMs and } an SEM as well. My question is, is their any facility that is COMPLETELY } cost neutral with revenue only for service work provided? Are the numbers } I describe for a medium sized University reasonable? I would greatly } appreciate any input. If their is a University that spends only what they } bring in for doing samples in a multi-user environment please contact me I } be happy to know how you did this. } } Best Regards, } } Kirk J. Czymmek, Ph.D.
This is what I've been faced with for the last four years. I was able to work out a deal with the enginnering school to fund a portion of my expenses (diminishing to ~$50K/year) for classroom activities (lecture/lab for a variety of courses) with the remainder of the roughly $120K budget made up by direct charge-back to sponsored projects. I've been able to make it so far (barely), but my biggest problem is equipment replacement (which we all know is very expensive!). I think I'm currently losing some high end work to other installations with better equipment, but I hope to upgrade soon.
For reference I have these income components for the lab: Educational mission Sponsored project charge-back Unsponsored projects (gets taken from educational mission funds) Off campus "clients"
I'd like to secure some "harder" monies but those days are all but gone.
Let me know if you can come up with a better way...
Brian
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax) "Be well, do good work, and keep in touch"
I have a Philips 300 that I need to sell. She has been my teaching scope for ten years but non-the-less well cared for. It has the DP and backing tank up grade as well as a plethora of spare parts.
No reasonable offer will be refused... the esteemed Dr. Mark Farmer from the University of Georgia finished up his thesis work with this instrument, so just think of its historical value alone!
The instrument is located in the Electron Imaging Facility at Rutgers University. Please contact either through E-mail or phone {732-445-5308 est}
Wet, dreary, traffic, strange smells, thats life in the Garden State.
John Grazul Rutgers University Electron Imaging Facility
The 2-methyl butane is actally the cryogen (freezing agent)and the LN2 is what is used to bring it down the the proper temperature. The reason that agents like 2-methyl butane, propane, ethane and freons etc. are used is that they have a higher boiling point than LN2 (-196 oC). LN2's low boiling point creats a vapor barrier (Leidenfrost phenomena) to form around tissues that are immersed directly into it which prevents the rapid conduction of heat from the tissue to the cryogen. The best cryogens have high boiling points and and low freezing points.
hope that is the answer to you question
-- Begin original message -- } } Could someone tell me what exactly is the role of 2-methyl butane when used } in } snap freezing tissue in liquid nitrogen? I appreciate any insight ... } } Andrea Hooper } Dept of Pathology } NYU Medical Center
-- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
We are doing ICC of unfixed cell bits (synaptic densities) stuck to glass. In spite of blocking with bacitracin and BSA (after sticking the bits to glass freshly cleaned with conc nitric acid), there is a scattering of colloidal gold on the glass (but virtually no stuck gold with the seconday/gold conjugate alone). I am wondering if anyone has come across a blocking or glass cleaning regime that can be applied after sticking the cell bits to the glass that will inhibit the primary antibody from sticking to the glass and keep background under control. Many thanks....Tom Reese
A few hours after posting this last week we had a major power failure and our server was out many days. I have had some complaints of unavailability, and repost. Please excuse any inconvenience. ****************************************** Fellow list members, BibMic - A bibliography of books relating to Materials Microscopy, previously published on paper in Materials Characterization 36(1996)105 is now available on the net at http://bibmic.metalmat.ufrj.br It lists over 1000 books, and is searchable by author, title and keywords. Hope you find it useful Prof. Walter A. Mannheimer Dept. of Metallurgy and Materiais Eng. Federal University of Rio de Janeiro POBox 68505, 21945 Rio de Janeiro, Brazil Vox (55 21) 590-0579 Fax (55 21) 290-6626 wamann-at-metalmat.ufrj.br
I have lost track of the vendor for my favorite 3 1/4 X 4 1/4 negatives sleeves. The package has the name RPS, catalogue # S-00650, Made in Japan, Plastine Film Preservers, Acid Free. I would appreciate learning who distributes these. I am not interested in any of the other brands or types. Thank you.
} } Could someone tell me what exactly is the role of 2-methyl butane when used in } snap freezing tissue in liquid nitrogen? I appreciate any insight ... } } Andrea -
I presume you're talking about plunge freezing. If you do this directly into liquid N2, the N boils, producing a layer of gaseous N around the tissue. That's a good insulator, so you get slow freezing. So you use an intermediate coolant that will stay liquid up to ~-80, for rapid heat transfer. When I was still in the lab (I'm retired) we used Freon 22, which is no longer permitted, or propane, which explodes if you're careless.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Dear Henk, } } I believe that TEM film has a density range of about 2.8-2.9. If this is } true, any scanner with a range of over 3 should work. } This is true for most EM images; however, ED patterns can have higher OD's. For quantitating ED intensities it is better to have as great a dynamic range as possible. Yours, Bill Tivol
Does anyone know of a book or a web site that is essentially an atlas of bacterial ultrastructure. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Assistant Director, The Biotechnology Program PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Responding to the message of {3.0.2.32.19980423103550.00701d5c-at-itsa.ucsf.edu} from Larry Ackerman {mishot-at-itsa.ucsf.edu} : } } } I have lost track of the vendor for my favorite 3 1/4 X 4 1/4 negatives } sleeves. The package has the name RPS, catalogue # S-00650, Made in Japan, } Plastine Film Preservers, Acid Free. I would appreciate learning who } distributes these. I am not interested in any of the other brands or types. } Thank you.
Larry, RPS (Reeves Photo Sales, Inc.) is at 9000 Sovereign Row, Dallas, Texas 75247, as of 3-4 years ago. I don't have their phone number. They also make really nice 35mm negative jackets.
Good Luck,
Gib
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
We are looking at some cells grown on Falcon cell culture inserts (essentially polyethylene terephthalate membrane filters). We fixed and osmicated, embeded in an epon-like epoxy resin. When we cut 0.5 um thick semi-thin sections, they wrinkle up something fierce due to the membrane. This makes it impossible to get a good LM photo. Anybody have any tricks? Falcon's tech bulletin was not very informative.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
We are planning on conducting some elemental analyses of 400-year-old glass beads. Would ZAF, Proza or Bence-Albee be the preferred correction method? We have a Noran Voyager with v2.7 software.
TIA
Bob Wise
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
This centrifugation/fixation works. We do this then cut the slender tube with a razor blade above and below the buffy coat (on a piece of Parafilm), making a short log with open ends. Then with a paper clip or applicator stick (depending on the diameter of the tube) we push out the packed buffy coat. If it tends to fall apart, we then encase it in 1% molten agar to keep it together. (You can encase it without washing out the glut, but don't fix the agar-encased pellet in glut of the other solutions won't infiltrate properly. If it sticks together, you can skip the agar. We then process the pellet or encased cells as a piece of tissue.
S Miller
On Thu, 23 Apr 1998, James Ito wrote:
} Date: Thu, 23 Apr 1998 09:29:17 +0100 } From: James Ito {james.ito-at-dial.pipex.com} } To: Microscopy-at-sparc5.microscopy.com, } Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } Subject: Re: COLLECTING A BUFFY COAT FOR TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We do this occasionally in my lab. } } We spin the anti-coagulated blood in a narrow test tube, then carefully } remove as much as possible of the plasma without disturbing the buffy } coat. Buffered 2% Glutaraldehyde is then very gently layered on top and } the tube left to stand in the fridge for about a couple of hours. This } gives a buffy coat which is embedded in solid plasma and can be removed } from the tube with the help of a thin wooden stick or similar. The } resultant disc can then be trimmed and the pieces processed to resin as } you would for normal tissue. If you embed in flat moulds your specimen } will have orientated layers of plasma, platelets, white cells and red } cells. } } It's a simple and effective technique, with only one centrigugation to } give you a sample which is easily handled. The only downside is that } some drugs (e.g. aspirin, I believe) may inhibit the action of } glutaraldehyde on the plasma and prevent the conversion of the plasma } to a solid. } } } James Ito } Pathology Department } Royal Hospital for Sick Children } Yorkhill } Glasgow } Scotland. } ---------- } } From: Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } } To: Microscopy-at-Sparc5.Microscopy.Com } } Subject: COLLECTING A BUFFY COAT FOR TEM } } Date: 22 April 1998 21:50 } } } } Hi There, } } Looking for a good technique for making a pellet of buffy coat from } } normal blood. Would appreciate any help. Thanks. } } } } Winnie } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
On Wed, 22 Apr 1998 hoopea01-at-endeavor.med.nyu.edu wrote:
} } Could someone tell me what exactly is the role of 2-methyl butane when } used in snap freezing tissue in liquid nitrogen? I appreciate any } insight ... }
Replies received so far are absolutely correct and on the nail. So I'm simply adding a couple of details for the general readers ...
(1) Being old fashioned, I still call 2-methyl butane ISOPENTANE, which might help if you're doing a literature search;
(2) If you have things that don't like being dunked in a hydrocarbon (rubbers and some plastics) it is possible to get quite powerful cooling by using a roughly equal mixture of acetone and methanol, which goes down to at least -110^C and has a higher boiling point than the isopentane. The b.p. of isopentane is twenty-something, which would make it hard to store in Arizona and similar places warmer than the UK.
Mixtures naturally have lower freezing points than the pure solvents, which is the principle of Dowtherm, an industrial heat transfer fluid which is mixture of Naphthalene and Diphenyl Ether.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Your blocking sems to be fine since gold conjugates do not stick. Have you tried to do the complete incubation on glass without cell bits on the surface? If you get the same background problem, you may have to purify or further dilute the primary. Are you using additives to the incubation buffer for the primary or only for the secondary? If there is no background on glass without cell bits, could it be that antigen leaks from the unfixed material?
Regards, Jan
============================= Jan Leunissen, Ph.D. AURION ImmunoGold Reagents & Accessories Managing Director Costerweg 5, 6702 AA Wageningen The Netherlands
---------- } From: wise-at-vaxa.cis.uwosh.edu } To: Microscopy-at-sparc5.microscopy.com } Subject: ZAF, Proza or Bence-Albee? } Date: April 23, 1998 7:42 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } To all, } } We are planning on conducting some elemental analyses of 400-year-old glass } beads. Would ZAF, Proza or Bence-Albee be the preferred correction method? } We have a Noran Voyager with v2.7 software. } } TIA } } Bob Wise } } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } www.uwosh.edu/departments/biology/wise/wise.html } } I understand Bence-Albee is the best way to go for minerals, and glass (old or not) is essentially a mineral, so there you go.
Quite right, but now the discussion goes to: Which is the better cryoagent and that was a topic here a few months ago. Propane gas liquefied by cooling is a much, much better cryo-agent than is isopentane. Its easy to store in a lab a small gas cylinder with a blunt needle on a bit of tubing as the outlet. With little gas flow rub the needle over the small metal cup that is cooled by liq N2. Soon you will have a couple of ml of liquid propane. Do this in a fumehood, which is a good idea when using solvents too. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** } } On Wed, 22 Apr 1998 hoopea01-at-endeavor.med.nyu.edu wrote: } } } } } Could someone tell me what exactly is the role of 2-methyl butane when } } used in snap freezing tissue in liquid nitrogen? I appreciate any } } insight ... } } } } Replies received so far are absolutely correct and on the nail. So I'm } simply adding a couple of details for the general readers ... } } (1) Being old fashioned, I still call 2-methyl butane ISOPENTANE, which } might help if you're doing a literature search; } } (2) If you have things that don't like being dunked in a hydrocarbon } (rubbers and some plastics) it is possible to get quite powerful cooling } by using a roughly equal mixture of acetone and methanol, which goes down } to at least -110^C and has a higher boiling point than the isopentane. } The b.p. of isopentane is twenty-something, which would make it hard to } store in Arizona and similar places warmer than the UK. } } Mixtures naturally have lower freezing points than the pure solvents, } which is the principle of Dowtherm, an industrial heat transfer fluid } which is mixture of Naphthalene and Diphenyl Ether. } } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: | } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } | University of Reading {University internal extension 7867 | } | Whiteknights Fax +44 (0) 118 9750203 | } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } | England URL: http://www.reading.ac.uk/~spsolley | } +------------------------------------------------------------------------+ } } }
Tom I have had similar problems with membrane filters of all kinds. I assume its due tho the differences in compression and stretching of the two materials. I did'n get rid of the wrinkles entirely but by embedding in a flat mold with the sample elivated off the bottom so that there was resin nearly equal above and below, the sections laid flatter and straighter.
Rick Vaughyn
PS Sometimes changing the hardness of the resin also helped, usually harder.
Dear Bob, I would try both and see if they differ. Generally, if there are heavier elements around, the proza correction is better at correcting the matrix absorbtion correction for the light elements. I have no experience with the Bence-Albee. It is very useful, with glasses, to run a well-characterized standard glass first, and try the different methods. You wrote:
} } To all, } } We are planning on conducting some elemental analyses of 400-year-old glass } beads. Would ZAF, Proza or Bence-Albee be the preferred correction method? } We have a Noran Voyager with v2.7 software. } } TIA } } Bob Wise } } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } www.uwosh.edu/departments/biology/wise/wise.html Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
Frank writes ... } } } } I understand Bence-Albee is the best way to go for } minerals, and glass } (old or not) is essentially a mineral, so there you go. }
Bence-Albee was a data reduction method for older computers ... e.g., a DEC PDP-5 with 4kb memory. It was a look-up table method of reducing elemental x-ray intensities to wt% oxides in the context of oxides. It was hardly computer intensive ... and many assumptions were made regarding simple hyperbolic modeling of the look-up beta factors. These assumptions hold true for select groups of elements, but many problems exists with the hyperbolic model. The other thing to realize is that the look-up table was created with ZAF ... that is, ZAF correction factors best fitted to the hyperbolic model. This would imply you'd do better with ZAF and no hyperbolic assumptions. Now ... lets talk about which "ZAF" ...
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
I notice that propane is actually somewhat soluble in (liquid) water, and I wonder if it diffuses into the sample to cause artifacts, perhaps altering membranes or extracting lipid droplets, which are my main interest. I would assume that IF this happens, isopentane being less soluble in water would have less of this kind of effect. Has anyone noticed?
In what way is propane noticeably better? Just due to lower temp? ( I must have missed the discussion a few months ago)
Thanks Richard
} } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } Quite right, but now the discussion goes to: Which is the better cryoagent and that was a topic here a few months ago. Propane gas liquefied by cooling is a much, much better cryo-agent than is isopentane.
... } } (2) If you have things that don't like being dunked in a hydrocarbon } (rubbers and some plastics) it is possible to get quite powerful cooling } by using a roughly equal mixture of acetone and methanol, which goes down } to at least -110^C and has a higher boiling point than the isopentane. } The b.p. of isopentane is twenty-something, which would make it hard to } store in Arizona and similar places warmer than the UK. } } Mixtures naturally have lower freezing points than the pure solvents, } which is the principle of Dowtherm, an industrial heat transfer fluid } which is mixture of Naphthalene and Diphenyl Ether. } } } +------------------------------------------------------------------------+ } | Robert H.Olley
If you happen to have very little blood you can also use microhematocrit tubes. The tube is broken near the buffy coat and the cut tip immersed in glutaraldehyde until the buffy-coat hardens. It can then be removed and processed. For details of the method see: Moura Nunes. J.F., Soares, J.O. and Alves de Matos, A.P. - Micro-Buffy Coats of whole blood: a method for the electron microscopic study of mononuclear cells. Stain Technol. 54(5)257, 1979
A.P. Alves de Matos mtlopes-at-fc.ul.pt
---------- } From: Sara Miller {saram-at-acpub.duke.edu} } To: James Ito {james.ito-at-dial.pipex.com} } Cc: Microscopy-at-sparc5.microscopy.com; Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } Subject: Re: COLLECTING A BUFFY COAT FOR TEM } Date: sexta-feira, 24 de abril de 1998 2:03 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } This centrifugation/fixation works. We do this then cut the slender tube
} with a razor blade above and below the buffy coat (on a piece of } Parafilm), making a short log with open ends. Then with a paper clip or } applicator stick (depending on the diameter of the tube) we push out the } packed buffy coat. If it tends to fall apart, we then encase it in 1% } molten agar to keep it together. (You can encase it without washing out } the glut, but don't fix the agar-encased pellet in glut of the other } solutions won't infiltrate properly. If it sticks together, you can skip
} the agar. We then process the pellet or encased cells as a piece of tissue. } } } S Miller } } On Thu, 23 Apr 1998, James Ito wrote: } } } Date: Thu, 23 Apr 1998 09:29:17 +0100 } } From: James Ito {james.ito-at-dial.pipex.com} } } To: Microscopy-at-sparc5.microscopy.com, } } Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } } Subject: Re: COLLECTING A BUFFY COAT FOR TEM } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } We do this occasionally in my lab. } } } } We spin the anti-coagulated blood in a narrow test tube, then carefully } } remove as much as possible of the plasma without disturbing the buffy } } coat. Buffered 2% Glutaraldehyde is then very gently layered on top and } } the tube left to stand in the fridge for about a couple of hours. This } } gives a buffy coat which is embedded in solid plasma and can be removed } } from the tube with the help of a thin wooden stick or similar. The } } resultant disc can then be trimmed and the pieces processed to resin as } } you would for normal tissue. If you embed in flat moulds your specimen } } will have orientated layers of plasma, platelets, white cells and red } } cells. } } } } It's a simple and effective technique, with only one centrigugation to } } give you a sample which is easily handled. The only downside is that } } some drugs (e.g. aspirin, I believe) may inhibit the action of } } glutaraldehyde on the plasma and prevent the conversion of the plasma } } to a solid. } } } } } } James Ito } } Pathology Department } } Royal Hospital for Sick Children } } Yorkhill } } Glasgow } } Scotland. } } ---------- } } } From: Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } } } To: Microscopy-at-Sparc5.Microscopy.Com } } } Subject: COLLECTING A BUFFY COAT FOR TEM } } } Date: 22 April 1998 21:50 } } } } } } Hi There, } } } Looking for a good technique for making a pellet of buffy coat from } } } normal blood. Would appreciate any help. Thanks. } } } } } } Winnie } } } } } } } } } Sara E. Miller, Ph. D. } P. O. Box 3020 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-8735
I am trying to find an etchant for deep etching a Ni-W eutectic alloy (between 17.5 + 20.7 at% W).
Also, I would appreciate any suggestions for electrically thinning such a material.
thanks in advance
Fred Pearson
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
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Richard: Isopentane becomes very rubbery near liquid nitrogen temperature. The temptation is to warm it a little with a metal rod, reducing the temperature differential some more. Propane has greater freezing speeds making vitrification possible. These things where published 20 and more years ago when cryo fixation was developed. I would be surprised if propane penetrates the specimen to any extent because snap freezing is so rapid. Later the specimen is sublimed and the propane would be the first phase to go. Also propane has been used to widely that somebody would have noticed an effect on lipids - if that was greater than isopentane's. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** } I notice that propane is actually somewhat soluble in (liquid) water, and I } wonder if it diffuses into the sample to cause artifacts, perhaps altering } membranes or extracting lipid droplets, which are my main interest. I } would assume that IF this happens, isopentane being less soluble in } water would have less of this kind of effect. Has anyone noticed? } } In what way is propane noticeably better? Just due to lower temp? } ( I must have missed the discussion a few months ago) } } Thanks } Richard } } } } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } } Quite right, but now the discussion goes to: Which is the better cryoagent } and that was a topic here a few months ago. } Propane gas liquefied by cooling is a much, much better cryo-agent than } is isopentane. } } ... } } } } (2) If you have things that don't like being dunked in a hydrocarbon } } (rubbers and some plastics) it is possible to get quite powerful cooling } } by using a roughly equal mixture of acetone and methanol, which goes } down } } to at least -110^C and has a higher boiling point than the isopentane. } } The b.p. of isopentane is twenty-something, which would make it hard } to } } store in Arizona and similar places warmer than the UK. } } } } Mixtures naturally have lower freezing points than the pure solvents, } } which is the principle of Dowtherm, an industrial heat transfer fluid } } which is mixture of Naphthalene and Diphenyl Ether. } } } } } } +------------------------------------------------------------------------+ } } | Robert H.Olley }
"New Developments in Multi-photon Excitation Microscopy"
to be held, July 11 - 12, in Atlanta GA, prior to the MSA annual meeting
Organizer: Jim Pawley
Cost: Workshop $300 (all day, HANDS ON in PM) Symposium $50 (lectures only)
Multiphoton excitation microscopy is the latest wrinkle in 3D light microscopy. Compared to earlier methods such as widefield deconvolution and confocal microscopy, multiphoton excitation promises a number of advantages: reduced effects of specimen-induced light scatter, better dichroic efficiency and the confinement of the photodamage zone to only the plane of focus. These improvements add up to the ability to image features far below the surface of thick, transparent biological specimens and to do so with up to 1000x less phototoxicity and photodamage than is produced by "normal" confocal.
With 3D light microscopy being used increasingly to follow developments in living cells, embryoes and tissues and the recent introduction of several commercial multi-photon instruments, this field is displaying explosive growth. This Symposium will include not only talks by experts from all the leading labs in the field but will also include contributions of some who have just begun: people like you who have overcome the technical challenges of "femtosecond" lasers to get outstanding results!!
In addition, there will be talks by experts in fluorescent lifetime imaging, a techniques that makes even better use of the pulsed nature of the exciting light.
We expect to have at least 4 new, operating, multi-photon imaging workstations for use by those who sign up for the Workshop.
TO REGISTER, CONTACT
Annamarie Dowling, MSA Meeting Manager 7000 W. Southwest Highway Chicago Ridge, IL 60415 (708)361-6000 / FAX -6166 {MSA-at-tradeshownet.com}
AND SHE WILL SEND YOU AN APPLICATION FORM.
TENTATIVE PROGRAM
Saturday July 11, AM: The Foundations of Multiphoton
Dave Piston Vanderbilt University, "Two-Photon Excitation Imaging of In Vivo Glucose Metabolism"
Warren Zipfel Cornell University "TBA"
Dave Wokosin University of Wisconsin "Multi-mode Multi-photon Microscopy"
Vinod Subramaniam, Max Planck Institute, Goettingen "Multi-photon microspectroscopy and imaging of near-UV fluorophores by scanning near-field optical microscopy (SNOM)"
Sunney Xie Battelle Pacific Northwest Laboratory "Single-Molecule and Near-field Fluorescence Imaging with Two-Photon Excitation: New Damage Mechanisms"
Hans Gerritsen Utrecht University "Two-photon excitation fluorescence lifetime imaging."
Saturday PM: Manufacturer's presentations I
Saturday Evening: Reception.
Sunday AM: Doing it our way
Mark Cannell University of Aukland, NZ " Visualization and quantification of the transverse tubular system in Living cardiac cells by 2-photon microscopy."
Steve Potter, Cal Tech "Two photon time-lapse of dendritic spines"
Hadley Wilson Horch, Duke University "Setting up multi-photon microscopy: problems and solutions"
Montrose Johns Hopkins "Multi-photon imaging of pH and drug therapy in the gastrointestinal tract"
Rebecca Williams , Cornell University "TBA"
James Pawley University of Wisconsin "Photon efficiency and QE in 3D Microscopy"
FOR MORE INFO contact: James Pawley, 1117 Johnson Ave, Madison , WI. , 53706, USA, Tel: 608-263-3147, Fax: 608-265-5315 E-mail: jbpawley-at-facstaff.wisc.edu
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
Lou Ann -- *************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
Microscopy Home Page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/CSMS/csms.html
Personal Home Page: http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/LAM.html
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I think that in general with issues of x-ray safety, the worst should always be assumed - so, x-rays may leak from an SEM.
In addition, I think there is also an argument to be made that SEMs could be more dangerous the TEMs. The higher the energy of an x-ray, the more penetrating it is and the longer the path over which it will deposit its energy. 30 kV x-rays could deposit more energy, and hence cause more damage, in a human body than 100 kV x-rays which having a shorter wavelength will interact less. I don't know what x-ray energy level is most dangerous to humans (perhaps somebody can enlighten) but I think it is wrong to assume that the higher the energy the more dangerous it is.
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
I am afraid that Larry's reply could not be "wronger". Some people will not know better and take the misleading information on board.
A TEM, when compared with a SEM potentially is the greater radiation risk. An SEM with a maximum of say 30kV is insignificant compared with a 200kV instrument. The wavelengths of electrons is changed with accelerating voltage but the K,L,M etc. X-rays generated by these electrons are in fact defined by the minimum electron energy required for their production. Regardless of the electron beam used to generate, the most powerful Cu X-ray is 8.978keV (the eV required to generate defines these X-rays). The minimum energy to generate the most powerful gold X-rays is 80.724keV. Those X-rays have far greater penetrating power and are rather more likely to leak from a badly modified instrument. The maximum X-ray generation occurs when the eV of the beam is about 2.8x the energy of a particular X-ray. So a 200kV instrument can almost reach maximum production of the most powerful K alpha Au X-rays. If only Al and C were used in an instrument (I know its not on) we would not generate such powerful X-rays. Unfortunately the light elements have very little X-ray stopping power. As a consequence in high kV instruments the manufacturers use in the very top aperture position an array of large, apertures with large, angled edges to stop the stray electron beam and the X-rays. Three or more apertures of different metals are used in that array to stop excess X-rays and to produce a "clean" beam for X-ray analyses. To do any harm X-rays must pass through the column. Happily that is easier prevented in the design of TEM and SEM than in X-ray diffractometers, which have far more movement of main components required. Any modern SEM or TEM, as provided by the manufacturers has so little leakage that microscopists should worry about the radiation received during a single flight at high altitude and not about sitting in front of these instruments. If you must modify an instrument, I suggest that an SEM is less likely to cause real grief than a TEM. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
-----Original Message-----
Postdoctoral/Senior technician opening:
An IMMEDIATE research position is available in the Department of Cell Biology, Baylor College of Medicine, Houston, TX, to join a team studying subcellular localization of steroid receptors and their cofactors, with a keen interest in nuclear/mitotic architecture. Moreover, particular interest will be centered upon covalent post-translational processing of wild-type and disease-associated receptors/cofactors in pathways leading to their turnover. In both cases, extensive effort will be focused upon live cell imaging using bioluminescence in multiple channels. High resolution studies at the ultrastructural level will focus on immunogold labeling and resinless-section microscopy. The successful candidate will have either/or extensive experience in cellular imagining, both by light and electron microscopic methodologies, strong skills in molecular cloning techniques and excellent communication skills (written and oral). This position can be appointed at the post-doctoral level, or as senior technician (BS/MS and } 7 years relevant experience). The annual salary range will be 25-30K, commensurate with experience.
These studies will make extensive use of the Department's Integrated Microscopy Core and the Computational and Visualization Biology Laboratory, containing state of the art hardware and software, including a new deconvolution-based wide-field optical workstation. High-speed data throughput is provided by a SGI Origin 2000 (equipped with 24 R10000 processors and } 1GB RAM), thus allowing nearly 'live' deconvolution imaging of four channels. Electron microscopy is supported by two TEMs, three ultramicrotomes and ancillary equipment. The Department offers a dynamic and intensive research environment with excellent opportunity for advancement.
Baylor College of Medicine is an Equal Opportunity/Affirmative Action/Equal Access Employer.
Interested applicants should forward (preferably electronically) a CV, letters of reference and research interests to:
Michael A. Mancini, Ph.D. Assistant Professor and Director Integrated Microscopy Core Department of Cell Biology Baylor College of Medicine Houston, TX 77030 mancini-at-bcm.tmc.edu 713 798 8592 713 790 0545 (fax)
Does anybody know if any or all the short courses at the M&M98 are still open? Or can anyone give me a phone contact? thank you Content-Type: image/gif Content-ID: {002401bd7146$92c6e680$6a8bd9ce-at-default}
I would think by now, most people on the Microscopy Listserver would know to go to the MSA WWW Site to find out information about M&M 98. But is seems that may not be the case. Here is the URL
http://www.msa.microscopy.com
To register or get information about the meeting contact the M&M98 Meeting Manager.
Contact information for both the Business Office as well as the Meeting Manager are also listed on the WWW site.
Microscopy & Microanalysis Meeting Manager
The Rebedeau Group 7000 W. Southwest Highway Chicago Ridge, IL 60415 Tel:(708) 361-6000; Fax (708) 361-6166 E-mail: MSAMeetingManager-at-MSA.Microscopy.Com
Mark Tobin wrote: ============================================ Has anybody heard of a material called Collodion being used for mounting materials for microscopy? I was asked by a colleague today and have not heard of it.
Any information, UK suppliers in particular, would be greatly appreciated == ========================================== Collodion™ is manufactured by nitrating with nitric acid and sulfating with sulfuric acid relatively ordinary cotton to make it soluble (as a result of the addition of soluble nitrate and sulfate groups). When dried, the solids become what is known as "gun cotton" because of its use as gun powder . We believe that Parlodion™ is very similar but it does have a different corporate and process origin and it unlikely to be exactly identical. At the very least, one could expect that the degree of nitrating and sulfating might vary between the several different manufacturers, thereby resulting in at least some subtle variation in the final properties. This could of course help explain why one researcher might have difficulty repeating someone else's work. However, most researchers report obtaining similar results, whether the application is for the casting of a film for TEM grids or for the making of replicas on metallurgical or ceramic types of surfaces.
We believe Collodion to be similar but not necessarily identical to the products called "Celloidin" and also, "LVN" , or "low viscosity nitrocellulose". And all of the mentioned products are supposed to be less explosive than the original "gun cotton".
These materials, e.g. all of the above mentioned products based on nitrocellulose nitrate, at one time, were also used widely as embedding resins, but have been replaced in most applications by more modern materials. However, for the embedding of really large samples, a technique called "double embedding" still requires a nitrocellulose type material. However, for specific kinds of samples, there might be a preference for one or the other of the nitrocellulose based materials.
Collodion, as well as the sister nitrocellulose based materials are flammable solids and require great care in their handling.
Disclaimer: SPI has offered both Parlodion and Collodion in both solid form as well as in solutions of 2% in amyl acetate for a long time. Both the solid material or solutions can be ordered either directly from SPI in the USA or from our distributor in the UK listed on our website. Our interest is to make sure that people understand that generically, these materials seem to be similar and while it is to the first approximation, if you have one "in hand", then you already have something that is very close to the other. Our other interest is to remind everyone that these are trade names of manufacturing companies and first use on a page should include the use of a "TM".
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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} I am afraid that Larry's reply could not be "wronger". Some people will not } know better and take the misleading information on board. } } A TEM, when compared with a SEM potentially is the greater radiation risk. } An SEM with a maximum of say 30kV is insignificant compared with a 200kV } instrument. The wavelengths of electrons is changed with accelerating } voltage but the K,L,M etc. X-rays generated by these electrons are in fact } defined by the minimum electron energy required for their production. } Regardless of the electron beam used to generate, the most powerful Cu X-ray } is 8.978keV (the eV required to generate defines these X-rays). The minimum
snips ..
} If you must modify an instrument, I suggest that an SEM is less likely to } cause real grief than a TEM. } Cheers } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } Great microscopy catalogue, 500 Links, MSDS, User Notes
Which actually misses the point I was trying to make:) I don't think that at any point I said that a TEM was safer than an SEM with regard to x-ray emissions. What I said, and the point that I was making, is that just because a SEM potentially generates less energetic x-rays than a TEM, it cannot be assumed that it is safer than a TEM (as Jim suggests).
As Jim points out, a SEM is just as capable as a TEM of generating plenty of x-rays. However, just because the HV is lower, it doesn't mean it is less dangerous. In addition, if people think that because of the lower HV, it is safer to modify SEMs, they will be heading for trouble - mainly because most people make an elementary mistake with regard to shielding for x-rays.
Many people think that to stop x-rays, you need a nice heavy metal, like lead. Which is correct as far as it goes but ignores the generation mechanism - electrons. Electrons, even at high kV have about as much penetrating power as a mosquito - high energy but no momentum. So, if you put a heavy metal in the way of an electron beam, it stops the electrons and generates lots of nice, highly penetrating heavy metal x-rays. The correct way to shield is a double layer of light elements and heavy elements - if electrons are going to hit anything, make sure it is aluminium or carbon. THEN, you have the lead to stop the Al x-rays - much safer and less lead needed.
The final point that I was raising is that not all x-rays are equally dangerous to people. I don't know the detail on this and enlightenment from someone who does would be useful. The energy of an x-ray (its wavelength) determines how strongly it interacts with matter. It is that interaction which causes damage. Something that passes straight through without interaction won't cause any damage - I guess we all have pleny of high energy neutrinos passing through us every day - with minimal effect. From this, very high energy x-rays, in themselves, would not be a direct danger. On the other hand low energy x-rays would be highly interacting and easily cause damage.
However, this observation need quantification - which I can't give. It would seem that there is an x-ray energy level which would cause most damage to a human. Below AND above that energy level would both be less dangerous. Now, IF that maximum danger energy is around 10 kV, SEMs will certainly be just as dangerous as TEMs. However, if the maximum danger energy level is 80 kV, then TEMs are a significantly greater potential danger.
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
{ Mark Tobin wrote: ============================================ { Has anybody heard of a material called Collodion being used for { mounting materials for microscopy? I was asked by a colleague { today and have not heard of it.
I have not heard to use Collodion as mounting material. However it can be used in replication techniques. To find out about this method you can read for instance:
D.H.Kay Techniques for E.M. (pages 60/61) G.Thomas Transmission E.M. on Metals (pages 134 & 179) A.M.Glauert in Practical Methods in E.M. Replica, Shadowing and Freeze-Etching Techniques (page 125)
Hans Brinkies Senior Lecturer SWINBURNE, University of Technology School of Engineering and Science Hawthorn, 3122, Melbourne - Australia Hbrinkies-at-swin.edu.au
Collodion is a 5% (I think) solution of celloidin in absolute alcohol and ether(diethyl). It is sold at this concentration for a number of simple applications. Fisher Scientific sells it, cat.#C408-500.
Scanning Probe Microscopy (SPM) Master Class Center for Interfacial Engineering University of Minnesota
DATE
Friday, May 22, 1998 and Saturday, May 23, 1998 Early registration deadline: April 24, 1998 Final registration deadline: May 8, 1998 Maximum Number of Attendees - 18
SPEAKERS
Professor Stuart Lindsay, Arizona State University Professor Andrew Hillier, University of Virginia Dr. Greg Haugstad, University of Minnesota (See enclosed interests/topics)
FORMAT
Friday morning (lunch provided) - speakers Friday afternoon - 4 hours on 6 experimental stations with expert assistance Saturday morning - 4 hours on 6 experimental stations with expert assistance
LOCATION
117/119 Smith Hall and 12 Shepherd Labs, University of Minnesota, Minneapolis, MN
PRICE
• Academic: $95 for early registration, $105 for late registration • Industrial: $295 for early registration, $325 for late registration • Price includes talks, instrument time with assistance, lunch
PAYMENT
With registration; print and mail attached registration card (TIFF).
CONTACT INFORMATION
Call Characterization Facility at 612-626-7594 to reserve spot. Attendees must bring samples and have prior SPM experience
HOTELS
Radisson Hotel 1-800-333-3333 615 Washington Avenue S.E. Minneapolis, MN 55414
Metrodome Days Inn 612-623-3999 2407 University Avenue S.E Minneapolis, MN 55414
Holiday Inn Metrodome 1-800-448-3663 1500 Washington Avenue So. Minneapolis, MN 55454
ABOUT THE SPEAKERS:
Stuart Lindsay is professor of physics at Arizona State University and Vice President of Research and Development at Molecular Imaging, a scanning probe microscope manufacturer. He received his Ph.D. in physics from the University of Manchester in 1976. In addition to numerous original-research articles, he has authored 16 review articles/book chapters and received 6 patents. His research interests are in biological physics: primarily biomolecular structure and electron transfer processes, as well as the development of new SPM instrumentation to enable these studies. Specific topics include the binding of regulatory proteins that switch certain genes on and others off. Prof. Lindsay will speak about in-fluid scanning force microscopy including MAC mode (intermittent contact under magnetic cantilever modulation), DNA imaging, and stiffness measurements within ordered fluid layers
Andrew Hillier is assistant professor of chemical engineering at the University of Virginia. He received his Ph.D. in chemical engineering from the University of Minnesota in 1995. His research interests focus on the chemical engineering aspects of materials and interfaces. This involves a range of topics including electrochemistry, materials chemistry, crystallization, catalysis, and the development and application of high resolution imaging techniques for in situ characterization of interfacial processes. Specific topics include the study of intermolecular forces and ordering phenomena in molecular systems, particularly during the crystallization of pharmaceutical products. Prof. Hillier will speak about resolution limits in scanning force microscopy, double-layer force measurements in electrolyte, and AC imaging with a thermally-driven cantilever.
Greg Haugstad is research associate in the CIE Characterization Facility at the University of Minnesota. He received his Ph.D. in physics from the University of Minnesota in 1991. His research interests are in polymer physics, with primary focus on energy-dissipative phenomena such as friction and viscoelasticity. Subtopics include the response of polymers to perturbative processes in SPM, i.e. changes in molecular-scale structure and/or properties induced by local shear/tensile forces. Another subtopic examines the mechanical/adhesive character of single or countable polymer molecules. A common thread in all of his work is the development of SPM methodologies to study nanoscale phenomena. Dr. Haugstad will speak about temperature-dependent scanning force microscopy of polymer friction/wear, and dynamic mode on polymer films with amplitude and phase imaging.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
URGENT: Application Deadline extended to May 8, 1998. ADJUNCT FACULTY NEEDED FALL Semester 1998 (Aug. 12 - Dec 18, 1998)
A San Joaquin Delta College Faculty member of the Microscopy Technology = Center is planning a Sabbatical Leave for the Fall '98 semester. San = Joaquin Delta Community College is currently searching for an EM = Instructor on an Adjunct or Temporary One Semester Contract. The = Contract will be at the discretion of the district. MINIMUM QUALIFICATIONS: Bachelor's Degree plus two years of directly related experience OR an = Associate Degree plus six years of directly related experience OR a valid = credential. DESIRABLE QUALIFICATIONS: Master's or PhD Degree in a Biological = Science; Experience in teaching Electron Microscopy COURSES TO BE TAUGHT: Introductory Techniques for Transmission Electron Microscopy (EM21) This is a lecture/lab course which includes beginning Transmission = Electron Microscopy dealing with the alignment and operation of the TEM, = vacuum techniques, photographic techniques, as well as the preparation of = particles and replicas for viewing in the TEM. Includes individual = training in the use of the TEM, preparation techniques, and written and = oral reports. (Lec - 2 hrs; Lab - 3 hrs/wk) Biological Ultrastructure (EM28) Course contents include specific information about the fine structure and = function of cells and tissue at the ultrastructure level. Videos, slides = and micrograph examination will be correlated with the lectures so that = students will learn to recognize the fine structure of cells and tissues = in relationship to their function. (Lec-2 hrs/wk) Current Microscopies: Optics, Theory and Application (EM30) Course contents include information related to the physical laws and = applications of the various types of current microscopies e.g. = TEM,SEM,FIB, AFM, and confocal microscopy, as well as other current = topics e.g. asbestos analysis, lab design, etc. (Lec - 2 hrs; Lab - 3 = hrs/wk) Advanced Techniques in Biological Electron Microscopy (EM37) Course contents include lecture and laboratory which covers advanced = techniques for biological specimen preparation in TEM including an = advanced research project.( Lec - 1 hr; Lab - 6 hr/wks) TERMS OF EMPLOYMENT: Adjunct / Non-tenured track position. APPLICATION: Contact Human Resources at 209/954-5056. DEADLINE: The Screening Committee will begin to review applications on = May 12, 1998
SJDC Microscopy Technology Program Information available at = http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html/
Human Resources San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207
id KAA21448; Mon, 27 Apr 1998 10:56:38 -0600 (MDT) Received: from nestor.NMSU.Edu by NMSU.Edu (8.8.8/NMSU-1.18) id KAA07444; Mon, 27 Apr 1998 10:57:46 -0600 (MDT) Received: from Microscope.nmsu.edu (microscope.NMSU.Edu [128.123.5.85]) by nestor.NMSU.Edu (8.8.6/8.7) with SMTP id KAA07872 for {microscopy-at-sparc5.microscopy.com} ; Mon, 27 Apr 1998 10:56:32 -0600 Message-Id: {3.0.2.32.19980427105655.0068f194-at-cnmailsvr.nmsu.edu} X-Sender: rtindell-at-cnmailsvr.nmsu.edu X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.2 (32)
Hi,
We are analyzing airborne particulates in an ongoing project and one question that has arisen concerns the possibility of separating the sample from the filter matrix. One particular sample was collected on a small fiberglass-type filter, making it nearly impossible to get into the fibers with the probe and get decent EDS done on particles. We have tried to take an overall spectrum of the filter, then subtract out a spectrum taken from an unused filter under identical beam/time/etc. conditions, but we're working blind as far as knowing if these results are giving us a real picture of what's there.
Is there a way of treating these filters, perhaps with solvents and sonication, that would give separate out enough particles to mount on a stub? We will try some things here, but someone with previous experience might be able to save us considerable time. The idea is to get an "overall" composition of airborne contaminants from the site, with additional analysis of individual particles, so hopefully what we separate from the filter would be representative of what was really in the filter. (Aye, there's the rub...)
Hopefully yours, Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Browsing in (Merriam) Webster's Collegiate and Hackh's Chemical dictionary reveals many interesting tidbits, though not necessarily useful ones. Of possible interest: collodion and pyroxylin are mostly the tri and tetranitrate of cellulose, gun cotton (main ingredient of smokeless powder) the hexanitrate. Terms first used ca. 1850-1880. No hint that collodion is a trademark, though Celluloid and Cellophane are or were. Use of ethyl alcohol + ethyl ether as solvents was possibly at one time superseded by the Cellosolve family (R-CH2CH2-OH), which combines the ether and hydroxyl functions in one molecule, but this group of solvents is now considered rather toxic.
Leonard R. Corwin Fort Dodge Animal Health Cyanamid Agricultural Research Center Quaker Bridge & Clarksville Roads PO Box 400 Princeton, NJ 08543-0400 609-716-2278 609-275-5239 fax corwinl-at-pt.cyanamid.com
{fontfamily} {param} New_Century_Schlbk {/param} RESEARCH ASSOCIATE POSITION AVAILABLE
A Research Associate position is available immediately to work on the 3-D structure of muscle and muscle/cytoskeleton proteins. One project aims to determine the structure of crossbridges in contracting insect flight muscle using fast freezing/freeze substitution procedures which trap force bearing myosin crossbridges with millisecond time resolution. Electron tomography is then used to produce 3-D images that preserve the variable structure of the crossbridges for subsequent analysis. Mechanical records on stiffness and tension are recorded up to the moment of freezing. Parallel time resolved X-ray diffraction data of the diffrent contractile states is also available with which to compare the EM data with native muscle structure. This project offers a unique opportunity to learn electron tomography, correspondence analysis of 3-D motifs and atomic modeling. The project involves primarily computing to obtain the 3-D images and computer modeling to interpret the structure. =20
The second project involves structure of cytoskeletal proteins on lipid monolayers, among these are alpha-actinins from many different species and tissues. The alpha-actinin crystals are large in extent and to date yeild structural information to at least 10 Angstroms resolution.=20 In addition, we have obtained crystals of a wide range of other proteins as well. Project is funded through January 2001. This project offers the opportunity to learn protein crystallization on lipid monolayers and methods for producing multiprotein complexes for 3-D imaging. =20
Applicants for both positions must have a PhD degree. Salary and relocation funds are negotiable based on relevent experience. =20
The successful applicant will become a member of the Structural Biology program at Florida State University that includes 3 protein X-ray crystallography groups, three macromolecular NMR groups, 2 EPR groups and 2 electron microscopy groups. Facilities for electron protein crystallography include a Philips CM300-FEG, a Philips CM120, 3 Gatan cryotransfer systems, a Gatan wide angle TV camera, a Perkin-Elmer PDS1010M densitometer, 2 surveying optical diffractometers. Inquiries and applications should be made to Dr. Kenneth Taylor, Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306-4380. Tel: (850) 644-3357. FAX (850) 561-1406. E-mail:=20 taylor-at-bio.fsu.edu. Applicants should provide a CV and the names and addresses of 3 former mentors or knowledgeable individuals who can provided reference letters. =20
Recent Pertinent Publications:
K. A. Taylor & D. W. Taylor. Projection image of smooth muscle {/fontfamily} {fontfamily} {param} Symbol {/param} a {/fontfamily} {fontfamily} {par= am} New_Century_Schlbk {/param} -actinin from 2-D crystals formed on positively charged lipid layers.=20 {italic} J. Mol. Biol. {/italic} {underline} 230 {/underline} , 196-205 (1993).
K. A. Taylor & S. Varga. Similarity of 3-D microcrystals of detergent solubilized (Na {smaller} + {/smaller} ,K {smaller} + {/smaller} )-ATPase from pig kidney and Ca {smaller} 2+ {/smaller} -ATPase from skeletal muscle sarcoplasmic reticulum. {italic} J. Biol. Chem.=20 {/italic} {underline} 269 {/underline} , 10107-10111 (1994). =20
H. Schmitz, C. Lucaveche, M. K. Reedy & K. A. Taylor. Oblique section 3-D reconstruction of relaxed insect flight muscle reveals the crossbridge lattice in helical registration. {italic} Biophys. J. {/italic} {underline} 67 {/underline} , 1620-1633 (1994). =20
H. Winkler & K. A. Taylor. 3D reconstruction by combining data from sections cut oblique to different unit cell axes.=20 {italic} Ultramicroscopy {/italic} {underline} 55 {/underline} , 357-371 (1994). =20
K. A. Taylor & D. W. Taylor. Formation of 2-D complexes of F-actin and crosslinking proteins on lipid monolayers: demonstration of unipolar {/fontfamily} {fontfamily} {param} Symbol {/param} a {/fontfamily} {fontfamily} {par= am} New_Century_Schlbk {/param} -actinin-F-actin crosslinking. {italic} Biophys. J. {/italic} {underline} 67 {/underline} , 1976-1983 (1994)
H. P. Erickson, D. W. Taylor, K. A. Taylor & D. Bramhill. Bacterial cell division protein FtsZ assembles into protofilament sheets and minirings; structural homologs of tubulin polymers. {italic} Proc. Nat. Acad. Sci. USA. {/italic} {underline} 93 {/underline} , 519-523 (1996)
Holger Schmitz, Mary C. Reedy, Michael K. Reedy, Richard T. Tregear, Hanspeter Winkler, Kenneth A. Taylor. Electron tomography of Insect =46light Muscle in Rigor and AMPPNP at 23=B0C. {italic} J. Mol. Biol. {/italic} 264, 279-301 (1996)
H. Schmitz, M. C. Reedy, M. K. Reedy, R. T. Tregear, H. Winkler, K. A. Taylor. Tomographic 3-D Reconstruction of Insect Flight Muscle Partially Relaxed by AMPPNP and Ethylene Glycol. {italic} J. Cell Biol. {/italic} {underline} 139 {/underline} , 695-707 (1997)
Kenneth A. Taylor, Jinghua Tang, Yifan Cheng & Hanspeter Winkler. The use of electron tomography for structural analysis of disordered protein arrays. {italic} J. Struct. Biol. {/italic} {underline} 120 {/underline} , 372-386 (1997)
} Mark Tobin wrote: } ============================================ } Has anybody heard of a material called Collodion being used for mounting } materials for microscopy? I was asked by a colleague today and have not } heard of it. } } Any information, UK suppliers in particular, would be greatly appreciated } ============================================
Collodion, as a mounting and particle manipulation medium for light and electron microscopy as well as elemental analysis, is widely discussed in the print Particle Atlas as well as the more recent Particle Atlas Electronic Edition. The sections are too extensive to quote here, but a couple of excerpts may be helpful:
} From Chapter 3, Particle Handling Techniques:
"Another mounting medium is flexible collodion. A dilute solution of particles in amyl acetate plus collodion is spread on a glass slide and allowed to dry, leaving the particles suspended in a clear film. Even though the refractive index of this medium is low, it disperses particles well. This medium is used primarily if there are very small particles ( {5 micrometers) to remove for further analysis. Collodion permits easy removal of single, very small particles by cutting out small squares of film after location microscopically."
Later, in the same chapter:
"A good example of how collodion can be used to separate particles from a matrix is a recent problem solved for a color TV manufacturer. Color TV tubes were being rejected because of tiny particles that produced green spots on the TV screen. A particle could not be picked up directly because it rested on a {1 micrometer aluminum film with a soft layer of phosphor beneath. The slightest touch of the needle would break the aluminum film, and the particle would be lost in the phosphor layer beneath. A 1 mm drop of collodion plus solvent was placed on the contaminated area and allowed to harden. The aluminum film, thus strengthened, was then lifted with a tungsten needle dipped in water. This wetted and held the loose phosphor beneath so that the collodion-aluminum film could be lifted, placed on a glass slide and softened with a drop of amyl acetate. An examination in transmitted light revealed a tiny hole in the aluminum film, with a 5 to 10 micrometer contaminant in the center. The particle was readily separated for microprobe and x-ray analysis with a No. 3 tungsten needle and just enough amyl acetate to allow the particle to flow slowly in the collodion."
For more information on the Particle Atlas Electronic Edition, contact me off-list at sshaffer-at-microdatawrae.com .
Hello, I am in search of a good protocol for the negative staining and TEM of microtubules assembled in vitro in the presence of taxol. Specifically, I need to know the following: Concentration of MTs/taxol Fix used and concentration (if any) Time of fix Duration of sample application to grid Duration of uranyl acetate application to grid
Please cc response to my email address: rcm7-at-psu.edu
Thanks, Rich Moore Penn State University rcm7-at-psu.edu
#################################################### Rosemary Walsh Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:rw9-at-psu.edu ####################################################
Larry Stoter wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am afraid that Larry's reply could not be "wronger". Some people will not } } know better and take the misleading information on board. } } } } A TEM, when compared with a SEM potentially is the greater radiation risk. } } An SEM with a maximum of say 30kV is insignificant compared with a 200kV } } instrument. The wavelengths of electrons is changed with accelerating } } voltage but the K,L,M etc. X-rays generated by these electrons are in fact } } defined by the minimum electron energy required for their production. } } Regardless of the electron beam used to generate, the most powerful Cu X-ray } } is 8.978keV (the eV required to generate defines these X-rays). The minimum } } snips .. } } } If you must modify an instrument, I suggest that an SEM is less likely to } } cause real grief than a TEM. } } Cheers } } Jim Darley } } } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } Which actually misses the point I was trying to make:) I don't think that } at any point I said that a TEM was safer than an SEM with regard to x-ray } emissions. What I said, and the point that I was making, is that just } because a SEM potentially generates less energetic x-rays than a TEM, it } cannot be assumed that it is safer than a TEM (as Jim suggests). } } As Jim points out, a SEM is just as capable as a TEM of generating plenty } of x-rays. However, just because the HV is lower, it doesn't mean it is } less dangerous. In addition, if people think that because of the lower HV, } it is safer to modify SEMs, they will be heading for trouble - mainly } because most people make an elementary mistake with regard to shielding for } x-rays. } } Many people think that to stop x-rays, you need a nice heavy metal, like } lead. Which is correct as far as it goes but ignores the generation } mechanism - electrons. Electrons, even at high kV have about as much } penetrating power as a mosquito - high energy but no momentum. So, if you } put a heavy metal in the way of an electron beam, it stops the electrons } and generates lots of nice, highly penetrating heavy metal x-rays. The } correct way to shield is a double layer of light elements and heavy } elements - if electrons are going to hit anything, make sure it is } aluminium or carbon. THEN, you have the lead to stop the Al x-rays - much } safer and less lead needed. } } The final point that I was raising is that not all x-rays are equally } dangerous to people. I don't know the detail on this and enlightenment from } someone who does would be useful. The energy of an x-ray (its wavelength) } determines how strongly it interacts with matter. It is that interaction } which causes damage. Something that passes straight through without } interaction won't cause any damage - I guess we all have pleny of high } energy neutrinos passing through us every day - with minimal effect. From } this, very high energy x-rays, in themselves, would not be a direct danger. } On the other hand low energy x-rays would be highly interacting and easily } cause damage. } } However, this observation need quantification - which I can't give. It } would seem that there is an x-ray energy level which would cause most } damage to a human. Below AND above that energy level would both be less } dangerous. Now, IF that maximum danger energy is around 10 kV, SEMs will } certainly be just as dangerous as TEMs. However, if the maximum danger } energy level is 80 kV, then TEMs are a significantly greater potential } danger. } } -- } Larry Stoter } 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK } email: LPS-at-teknesis.demon.co.uk } Phone/Fax: +44 (0)1825 767967
Larry,
The main point that you have missed is that there is virtually nowhere for x-rays to escape from an SEM. The chief point of generation is the gun area and I have not seen any with less than about 1/2" of steel (often 301 stainless) that must be penetrated. 30keV won't do it.
By the time you get down to the chamber, you are generating very few x-rays.
You know the x-ray film that the dentist exposes through your cheek, Jaw and teeth in 1/25 to 1/10 second? He's using a 70keV 15-25mA source. I'm not sure what the target metal is, but it may be copper. When I use that film to align a wavelength spectrometer, I am exposing it at a distance of about a foot from the beam with an absorbed current of 100nA on copper at 30keV. (This is a very high current for an SEM. A typical current for EDS work is 200-400pA and high res imaging is 1-10pA.) exposure is on the order of 5-10 MINUTES!
Most SEMs don't have anything but steel around the chamber. Some have glass ports for specimen airlocks, but the HV is generally interlocked. I had a customer in Canada who made a Plexiglass port and often ran 12-18 hour EDS scans. He put a film badge inside the metal cover for this port and left it there for a month. He got no report of dosage back after turning in the badge.
Some day I plan to survey an SEM with a light element detector, then survey the color monitor that goes with the EDS. I'm betting that there is nothing detectable from the SEM and considerable emission from the monitor. Any takers?
TEMs ARE inherently dangerous because they CAN generate very hard x-rays that are difficult to contain, and you are looking directly at the impact point of a lot of very energetic electrons. The column is also far more complex, making it harder to shield.
You're correct about harder x-rays spreading their energy over a greater volume of tissue, but you miss the point that the soft x-rays can't leave the system. Your color TV at home is probably a lot more dangerous than any SEM I've ever seen.
Ken Converse Quality Images Delta, PA third party SEM service
} The main point that you have missed is that there is virtually nowhere } for x-rays to escape from an SEM.
want a bet:)
} The chief point of generation is the gun area
so what do all those electron streaming down the column do when they hit apertures, valves, etc. Create fluffy bunnies?
} and I have not seen any with less than about 1/2" of steel } (often 301 stainless) that must be penetrated. 30keV won't do it. } } By the time you get down to the chamber, you are generating very few } x-rays.
I'd agree that the greatest intensity of x-rays is likely to be generated around the gun - SEM or TEM. However, x-rays can be generated elsewhere. And remember this question related to modifications - if somebody starts making modifications anywhere they risk letting x-rays out.
} You know the x-ray film that the dentist exposes through your cheek, Jaw } and teeth in 1/25 to 1/10 second? He's using a 70keV 15-25mA source. } I'm not sure what the target metal is, but it may be copper. When I use
I know my dentist runs for cover everytime he x-rays my teeth. I can't help wondering as I sit there with the tube against the side of my head and the dentist 20 feet away behind a lead-lined wall if there is an international conspiracy by dentists to take over the world:)
} that film to align a wavelength spectrometer, I am exposing it at a } distance of about a foot from the beam with an absorbed current of 100nA } on copper at 30keV. (This is a very high current for an SEM. A typical } current for EDS work is 200-400pA and high res imaging is 1-10pA.) } exposure is on the order of 5-10 MINUTES!
But you keep assuming that everything is working correctly, in an unmodified SEM. I bet the guys at Three Mile Island just KNEW there was nothing to worry about.
} Most SEMs don't have anything but steel around the chamber. Some have } glass ports for specimen airlocks, but the HV is generally } interlocked. I had a customer in Canada who made a Plexiglass port and } often ran 12-18 hour EDS scans. He put a film badge inside the metal } cover for this port and left it there for a month. He got no report of } dosage back after turning in the badge.
Which I for one find very worryinging.
} Some day I plan to survey an SEM with a light element detector, then } survey the color monitor that goes with the EDS. I'm betting that there } is nothing detectable from the SEM and considerable emission from the } monitor. Any takers?
Try leaving your film badge on top of the TV at home over the week end. A standard SEM will certainly allow fewer x-rays to escape than any TV or monitor. However, we aren't talking about standard SEMs - we are talking about ones that users modify.
Which reminds me - if you ever get the chance, try a radiation check in a long haul airliner. I had a colleague who one took a flight from Germany over to California, to do some radiation checks on an instrument before it was shipped to Germany, He hand-carried his monitoring instruments in the cabin. In mid-Atlantic, the inside of the plane broke the German radiation exposure limits - not by just a little bit but by a factor of more than double.
} TEMs ARE inherently dangerous because they CAN generate very hard x-rays } that are difficult to contain, and you are looking directly at the } impact point of a lot of very energetic electrons. The column is also } far more complex, making it harder to shield.
Please, point out precisely where I said that TEMs weren't dangerous. I'm making a point that SEMs as well have the potential to be dangerous and that dismissing this danger because the kV is lower and won't harm you, might just be a triffle foolish. The english language is actually capable of greater subtleties than simple right/wrong arguments.
} You're correct about harder x-rays spreading their energy over a greater } volume of tissue, but you miss the point that the soft x-rays can't } leave the system.
In principle, always provided that the original manufacturers configuration has not been modified - which is where the discussion started.
} Your color TV at home is probably a lot more } dangerous than any SEM I've ever seen. } } Ken Converse } Quality Images } Delta, PA } third party SEM service
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
I wonder if somebody out there can help. We are looking for a power regulator board for a Cambridge S200 SEM, part number 852594. Leo can make one up for us, but it will take up to six months to deliver as it is an old model. Please contact us if you can supply.
Luc Harmsen Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Looking for source/supplier to get Freon 12. This is required for HT Tank } of JEOL TEM 2000FX. Any help will be greatly appreciated. } } Zia ur Rahman } Materials Characterization Facility, } University of Central Florida, } Orlando, Florida, } USA } } zur-at-mmae.engr.ucf.edu } Hi,
The original freon gas used in the gun and HT tank of JEOL microscopes is no longer used as it is a CFC and environmentally damaging. JEOL reccomend that it be replaced with SF6 (sulphur hexaflouride) gas and certainly contacted UK users with details of a modification to do this. The SF6 has to run at a slightly higher pressure: 0.18 to 0.20 kg/cm2 in the tank and 2.8 to 3.0 kg/cm2 in the gun for our 2010 on SF6 compared to 0.05 to 0.15 kg/cm2 in the tank and 1.75 to 1.95 kg/cm2 in the gun for freon. This just involves a change of the gas fittings on the tank to enable a SF6 cylinder to be used for the gas fill, however, the gun requires a change in gas fittings, a new (higher range) pressure gauge and replacing the overpressure valve (or fitting one if not fitted). You also need to be careful when checking for gas leaks as the pressure is higher then previously used, make sure that all pipes and fitting that you reuse are good enough for the higher pressure.
Either contact JEOL or sort it out yourselves, we have converted 2 JEOL 4000s and a 2010 successfully and will convert our 200CX when it next needs to be serviced in either the tank or gun. We have found that the characteristics of the HT have changed a little. If we have a flashover there is no longer a carbon track to be cleaned but the HT set prefers to be allowed to recover for a while before applying the maximum voltage again (usually 10 mins or maybe 1 hour for large flashover at 400KV).
Make sure that you read the safety data for SF6 gas, before you convert your machine, and take the appropriate local action. A local (to you) company makes a very good leak detector for freon or SF6. Leak-seeker model L-790a from CPS Products Inc. Hialeah, FL 33013 (Phone (305) 687-4121). (No interests in JEOL or CPS)
Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
We are analyzing airborne particulates in an ongoing project and one question that has arisen concerns the possibility of separating the sample from the filter matrix. One particular sample was collected on a small fiberglass-type filter, making it nearly impossible to get into the fibers with the probe and get decent EDS done on particles. ... Is there a way of treating these filters, perhaps with solvents and sonication, that would give separate out enough particles to mount on a stub? ---------------------------------------------------------------------------- ------------------------------- You mentioned that this is an ongoing project so I assume you will be taking more samples. I would highly recommend that further sampling for airborne particulate be done with polycarbonate membrane filters. Gelman and Nuclepore are two brand names that come to mind. The environmental services labs which monitored for asbestos used these extensively. The polycarbonate membrane is solid film with sub-micron, radiation induced holes etched in them which makes them ideal for generating carbon film replicas for TEM analysis. The particles of interest remained embedded in the carbon film while the filter was dissolved away. In contrast, larger particles can be analyzed directlyon the filter in the SEM or easily picked off of the filter surface.
Regarding your fiberglass filter, I'm sorry I can't help you too much. Whatever you know about the air particulate should guide you. Is it mineral or organic?
Dear Larry, Ken, et al., } } } The chief point of generation is the gun area } The chief points of generation in our high-voltage TEM are below the accelerator between the intersystem valve and the 1st condenser aperture.
} so what do all those electron streaming down the column do when they hit } apertures, valves, etc. Create fluffy bunnies? } Mostly, they create brehmsstrahlung (continuous energy spectrum) radiation.
} } and I have not seen any with less than about 1/2" of steel } } (often 301 stainless) that must be penetrated. 30keV won't do it. } }
The half-thickness for absorption is on the order of 100 mg/cm^2, which is about 0.1 mm of steel. A negligable fraction of 30 keV x-rays will get through 1 cm.
} And remember this question related to modifications - if somebody starts } making modifications anywhere they risk letting x-rays out. } The modification must involve a sufficiently short path so that a significant portion of the radiation can pass through. A relatively thin port would do the job, however.
} I know my dentist runs for cover everytime he x-rays my teeth. I can't help } wondering as I sit there with the tube against the side of my head and the } dentist 20 feet away behind a lead-lined wall if there is an international } conspiracy by dentists to take over the world:) } The major reason (s)he runs for cover is that (s)he exposes many x-ray films per day; whereas you are subjected to this procedure about once per year. In your case, the benefits outweigh the exposure risks, but the dentist receives no benefit from x-ray exposure, so trys to keep that exposure "as low as reasonably achievable". } } But you keep assuming that everything is working correctly, in an } unmodified SEM. I bet the guys at Three Mile Island just KNEW there was } nothing to worry about. } They were more-or-less correct. The guys at Chernobyl probably felt the same way, but were definitely incorrect. The estimated biolog- ical effect of Three Mile Island is about one excess cancer in the total exposed population. TMI was a disaster only to the utility company. Chernobyl was the real disaster with many people killed from acute radia- tion sickness and millions more exposed to significant radiation doses, which will cause latent effects including cancer and genetic abnormalities.
} } Most SEMs don't have anything but steel around the chamber. Some have } } glass ports for specimen airlocks, but the HV is generally } } interlocked. I had a customer in Canada who made a Plexiglass port and } } often ran 12-18 hour EDS scans. He put a film badge inside the metal } } cover for this port and left it there for a month. He got no report of } } dosage back after turning in the badge. } } Which I for one find very worryinging. } We have put thermoluminescent dosimeters, sensitive to millirad doses, around our microscope and have seen only background radiation. There is more radiation from the concrete in the building than from the microscope. Since we are always concerned about the effects of the modifications we have made over the years, we have taken care not to provide leakage pathways and have monitored this closely. However, lack of this sort of attention can, indeed, lead to situations where there are radiation leaks. Yours, Bill Tivol
Dear Larry, et al., } } } A TEM, when compared with a SEM potentially is the greater radiation risk. } } An SEM with a maximum of say 30kV is insignificant compared with a 200kV } } instrument. The wavelengths of electrons is changed with accelerating } } voltage but the K,L,M etc. X-rays generated by these electrons are in fact } } defined by the minimum electron energy required for their production. } } Regardless of the electron beam used to generate, the most powerful Cu X-ray } } is 8.978keV (the eV required to generate defines these X-rays). The minimum } Most of the x-rays generated are brehmsstrahlung x-rays, which have a continuous energy spectrum. A 200 keV electron can produce an x-ray with the same energy (it is stopped by a large mass--like a heavy atom--which conserves the momentum, and the energy is carried by the photon). The argument about characteristic x-ray generation, while correct, is not the whole story. Another complication is that characteristic x-rays are emit- ted isotropically, while brehmsstrahlung x-rays are mostly forward-directed. Since, however, electrons can be elastically scattered in various directions before producing brehmsstrahlung, the distrubution of such radiation produ- ced by a microscope depends on what the electrons are likely to hit as they go through the column.
} The correct way to shield is a double layer of light elements and heavy } elements - if electrons are going to hit anything, make sure it is } aluminium or carbon. THEN, you have the lead to stop the Al x-rays - much } safer and less lead needed. } Beryllium is also very good in this regard. One needs the lead to stop not only the Al x-rays, but the potentially more energetic brehms- strahlung x-rays.
} The final point that I was raising is that not all x-rays are equally } dangerous to people. I don't know the detail on this and enlightenment from } someone who does would be useful. The energy of an x-ray (its wavelength) } determines how strongly it interacts with matter. It is that interaction } which causes damage. Something that passes straight through without } interaction won't cause any damage - I guess we all have pleny of high } energy neutrinos passing through us every day - with minimal effect. From } this, very high energy x-rays, in themselves, would not be a direct danger. } On the other hand low energy x-rays would be highly interacting and easily } cause damage. } The half-thickness for the absorption of tens-of-keV x-rays in tissue is on the order of a centimeter. Any x-radiation which is pro- duced by a microscope will, therefore, penetrate far enough to lose most of its energy in the living part of a person, and would be almost comple- tely absorbed before passing through that person. A 1 MeV x-ray has a half-thickness of about 10 cm in tissue, so most of its energy will also be absorbed before passing through. These values come from a figure in Friedlander et al., Nuclear and Radiochemistry.
} However, this observation need quantification - which I can't give. It } would seem that there is an x-ray energy level which would cause most } damage to a human. Below AND above that energy level would both be less } dangerous. Now, IF that maximum danger energy is around 10 kV, SEMs will } certainly be just as dangerous as TEMs. However, if the maximum danger } energy level is 80 kV, then TEMs are a significantly greater potential } danger. } Unfortunately, the danger vs energy is a pretty flat curve with a broad maximum. Any scope (or other equipment) from which x-rays escape has the potential to be dangerous. All the more so since most of us work around the equipment for a large part of the day. Yours, Bill Tivol
could anybody send me an electronic version of the article "de Ruijter (1995) Imaging properties and applications of slow-scan charge-coupled device cameras suitable for electron microscopy, Micron, 26, 247-276" asap or indicate the web address where it can be found. Thanks a lot. With best regards,
Dmitry Cherny
Current address: MPI for Biophysical Chemistry, dept. of Molecular Biology am Fassberg 11, D-37077 Gottingen, Germany tel: +49(0) 551 201 1765; fax: +49(0) 551 201 1467 e.mail: dtcherny-at-mpc186.mpibpc.gwdg.de dtcherny-at-img.ras.ru
Wanted to put in my 2 cents worth before this thread closes. Obviously both sides have their points, and have made the larger, more important point, which is if you don't know how sharp the fangs and claws are, don't play with the cat. This goes for many pieces of equipment around the EM lab, not just limited to the electron microscopes but also including sputtering devices, ion mills, evaporators(electron beam especially), and critical point driers, to name only a few. All are capable of inflicting injuries as we have seen in the past. Repairs and modifications should be done by, or at least in consultation with experts such as Jim, Kenneth, Larry or the manufacturer, who recognize the dangers and can give advice to avoid them. A couple of additional things, I wanted to mention; 1. Any time that the high voltage exceeds 10 kV, regular glass is no longer capable of shielding against emitted x rays. Lead glass or metal is necessary. 2. It needs to be mentioned, although we all realize it, the ability to adequately detect radiation decreases as the energy decreases. A Geiger counter that is ok at 100kV does not usually truly indicate the magnitude of the radiation at 10 kV. Special equipment is needed at this level. I also understand film badges and dosimeters also fall off in their ability to detect low energy x-rays and special devices are needed. 3. Antidotally, I have never detected a radiation leak in a SEM but easily can find some area in most TEMs that will give some counts. (Just to add some perspective, you can usually detect some counts above background, around color computer monitors which usually operate around 30 kV and at about an order of magnitude higher current then SEM columns.) In the TEM's the radiation mostly emanates as beams. Usually the manufacturer covers these areas with lead shielding. This brings up the 4th item, make sure that all parts are returned to the microscope and in the proper position while doing repairs or cleaning. The innocent looking washer or plate may be shielding the manufactured included to prevent just these same radiation beams or leaks. Turning on a TEM without the lap shield in place or looking into the oil fill hole on top of an SEM gun while the HV is operating can subject you to unnecessary radiation exposure. We have several shields on an older microscope that can shift out of place easily during cleaning and need to be checked frequently.
Dear Randy, I don't know of any way to dissolve fiberglass, short of HF, so you should use a dissolvable filter in the first place. Nucleopore filters are my preference, since they are flat, as the particles sit up on top instead of down in the mesh and their EDS background is just polycarbonate (C and O). They can be dissolved, if necessary, in a Jaffe washer in chloroform for 48 hours. You wrote: } Hi, } } We are analyzing airborne particulates in an ongoing project and one } question that has arisen concerns the possibility of separating the sample } from the filter matrix. One particular sample was collected on a small } fiberglass-type filter, making it nearly impossible to get into the fibers } with the probe and get decent EDS done on particles. We have tried to take } an overall spectrum of the filter, then subtract out a spectrum taken from } an unused filter under identical beam/time/etc. conditions, but we're } working blind as far as knowing if these results are giving us a real } picture of what's there. } } Is there a way of treating these filters, perhaps with solvents and } sonication, that would give separate out enough particles to mount on a } stub? We will try some things here, but someone with previous experience } might be able to save us considerable time. The idea is to get an } "overall" composition of airborne contaminants from the site, with } additional analysis of individual particles, so hopefully what we separate } from the filter would be representative of what was really in the filter. } (Aye, there's the rub...) } } Hopefully yours, } Randy } } } Randy Tindall } Electron Microscope Laboratory } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } } rtindell-at-nmsu (work) } nrtindall-at-zianet.com (home) Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
Thanks for reminding us of the issue of brehmsstrahlung.
snips ...
} } However, this observation need quantification - which I can't give. It } } would seem that there is an x-ray energy level which would cause most } } damage to a human. Below AND above that energy level would both be less } } dangerous. Now, IF that maximum danger energy is around 10 kV, SEMs will } } certainly be just as dangerous as TEMs. However, if the maximum danger } } energy level is 80 kV, then TEMs are a significantly greater potential } } danger. } } } Unfortunately, the danger vs energy is a pretty flat curve with } a broad maximum. Any scope (or other equipment) from which x-rays escape } has the potential to be dangerous. All the more so since most of us work } around the equipment for a large part of the day. } Yours, } Bill Tivol
Which does add considerable weight to the point I was making - just because the kV of an SEM is lower than that of a TEM, it should not be assumed that it is less dangerous.
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
} Larry, } } The main point that you have missed is that there is virtually } nowhere for x-rays to escape from an SEM. The chief point of } generation is the gun area and I have not seen any with less than } about 1/2" of steel (often 301 stainless) that must be penetrated. } 30keV won't do it. } } By the time you get down to the chamber, you are generating very few } x-rays. } } You know the x-ray film that the dentist exposes through your cheek, } Jaw and teeth in 1/25 to 1/10 second? He's using a 70keV 15-25mA } source. I'm not sure what the target metal is, but it may be copper. } When I use that film to align a wavelength spectrometer, I am } exposing it at a distance of about a foot from the beam with an } absorbed current of 100nA on copper at 30keV. (This is a very high } current for an SEM. A typical current for EDS work is 200-400pA and } high res imaging is 1-10pA.) exposure is on the order of 5-10 } MINUTES!
Ken,
You and I come from the common point of ETEC SEMs. I try to use a wavelength alignment using 400nA, although that can be hard to achieve. At that point in the system, you are dealing only with a small subtended angle of the x-ray radiation emitted from the sample, after passing through a magnetic apeture that removes any electrons.
Regardless, consider the case where an ETEC has lost the small aluminum plug that covers the rectangular hole machined at the top of the objective lens. I would not leave that uncovered as it offers a path for a wide angle of x-rays emitted by the absorption of electrons in the electron-optics.
Regarding exposure to any ionizing radiation, such as x-rays, care must be taken to minimize long-term exposure to them. As an analogy, the current law suits brought by airline flight attendants regarding secondary tobacco should be seriously compromised by their constant exposure to the higher background radiation levels at the flight levels they encountered.
Ionizing radiation is an insidious problem, particularily at the energies present in an EM. It becomes not a problem of either-or, but a statistical problem of how-much, how-long. Normally unmeasured levels of radiation can over the long term produce potential problems.
Your later comments about the EDS monitor are correct. A typical survey of the equipment will show a substantial emission from the EDS monitor, and little or no emission from the SEM. However, you can not infer that this result means that there is no potential problem from the SEM.
Computer monitor manufacturers have recently realized the emissions from their equipment, and have taken steps to reduce them. SEM manufacturers have always been aware of the problems, and should have taken mechanical design steps to prevent a problem. But when those systems are being modified by the user, it is the user who must be aware of the problems inherent in those modifications.
} Most SEMs don't have anything but steel around the chamber. Some } have glass ports for specimen airlocks, but the HV is generally } interlocked. I had a customer in Canada who made a Plexiglass port } and often ran 12-18 hour EDS scans. He put a film badge inside the } metal cover for this port and left it there for a month. He got no } report of dosage back after turning in the badge. } } Some day I plan to survey an SEM with a light element detector, then } survey the color monitor that goes with the EDS. I'm betting that } there is nothing detectable from the SEM and considerable emission } from the monitor. Any takers? } } TEMs ARE inherently dangerous because they CAN generate very hard } x-rays that are difficult to contain, and you are looking directly } at the impact point of a lot of very energetic electrons. The } column is also far more complex, making it harder to shield. } } You're correct about harder x-rays spreading their energy over a } greater volume of tissue, but you miss the point that the soft } x-rays can't leave the system. Your color TV at home is probably a } lot more dangerous than any SEM I've ever seen. } } Ken Converse } Quality Images } Delta, PA } third party SEM service } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Hi - Perhaps I am missing something vital in my understanding of X-ray generation in EMs. Why should the bremsstrahlung be lower in SEM? I thought that bremsstrahlung is the result of electron interaction with the nucleus rather then electrons. A high continuum (versus defined X-rays) is typical of a spectrum generated by low atomic number elements. There is not much difference in the of elements used in SEM versus TEM.
Larry's argument that a SEM is potentially as dangerous as a TEM is at odds with my understanding of the subject and no valid argument has been advanced to change my view. I note that normally the various radiations are well contained in both instrument types, but consider what is happening within a 200kV TEM and a 30kV SEM.
The maximum energy of Pt (78; apertures) X-rays is 73.4keV and that of lead (82; shielding) is 88keV. It requires about 2.8x of the X-rays defining energy to maximise the production of those X-rays. In the 30kV SEM maximum emission of Zn (30; brass parts) with maximum of 10.4keV is possible; some emission of Mo (42; apertures) with maximum energy X-rays of 20.0keV. Clearly, a 200kV beam is sufficient to generate at lot of much more powerful - and I understand that these are also the most dangerous - X-rays.
If my understanding is wrong, still nothing would change to make the SEM as or more dangerous than the TEM. The TEM's powerful X-rays and primary and secondary electrons in turn exite other decreasingly powerful X-rays in a cascading reaction. Consequently the TEM not only generates powerful radiation within but also greater numbers of X-rays of numerous energy levels down to the ultra-soft.
As far as quantity of radiation is concerned: It is true that TEM's are commonly operated on 15 microamp emission and SEM at 80, but . . . . Without knowing actual numbers I am certain that a TEM operated at 200kV and 15uA produces not only rather more powerful X-rays but also greater total numbers of X-rays of all sorts of intensities when compared with a SEM operated at 30kV and 80 mA emission.
Many years ago I had to assume the additional role of an analyst. I am mostly self-tought in the relevant physics; I am happy to learn and not worried that I might be proven wrong. I think it is important that electron microscopists have some understanding of this topic. My conclusion is: If you are going to modify an SEM think and consult. If you are going to modify a TEM, think and consult twice. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** } Thanks for reminding us of the issue of brehmsstrahlung. } } snips ... } } } } However, this observation need quantification - which I can't give. It } } } would seem that there is an x-ray energy level which would cause most } } } damage to a human. Below AND above that energy level would both be less } } } dangerous. Now, IF that maximum danger energy is around 10 kV, SEMs will } } } certainly be just as dangerous as TEMs. However, if the maximum danger } } } energy level is 80 kV, then TEMs are a significantly greater potential } } } danger. } } } } } Unfortunately, the danger vs energy is a pretty flat curve with } } a broad maximum. Any scope (or other equipment) from which x-rays escape } } has the potential to be dangerous. All the more so since most of us work } } around the equipment for a large part of the day. } } Yours, } } Bill Tivol } } Which does add considerable weight to the point I was making - just because } the kV of an SEM is lower than that of a TEM, it should not be assumed that } it is less dangerous. } } Regards, } } -- } Larry Stoter } 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK } email: LPS-at-teknesis.demon.co.uk } Phone/Fax: +44 (0)1825 767967 } } }
We are attempting to prepare TEM cross-section samples of LaAlO3 single crystal subtrate with a few thousand angstroms of YBCO film deposited on the surface. So far we haven't been successful using ion beam thinning techniques.
Any help would be greatly appreciated.
Thanks in advance
Fred Pearson email: eoptics-at-mcmaster.ca
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
We've been very successful preparing YBCO on CeO2 on sapphire using the small-angle cleavage technique (see M.W. Denhoff and J.P. McCaffrey, "Epitaxial Y1Ba2Cu3O7 Thin Films on CeO2 Buffer Layers on Sapphire Substrates", J. Appl. Phys. 70 (7), p. 3986 (1991)). There is an excellent recent update of the technique in: S.D. Walck, J.P. McCaffrey, "The small-angle cleavage technique: an update", Mat. Res.
Soc. Symp. Proc. Vol.480, pp.149-171 (1997).
If you can't get a copy of the MRS proceeding, drop me a note offline and I'll send you some material plus our newest video on the technique, featuring Igor and Dr. Cleavinstein!
Cheers John
John P. McCaffrey Institute for Microstructural Sciences National Research Council of Canada M-50 Montreal Rd. Ottawa, Ontario K1A 0R6 Canada
Tel: 613-993-7823 Fax: 613-990-0202 email: john.mccaffrey-at-nrc.ca ---------------------------------------------------------------------------- -- REPLY FROM: McCaffrey, John (IMS) Microsoft Mail v3.0 (MAPI 1.0 Transport) IPM.Microsoft Mail.Note
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Fellow Microscopists:
We are attempting to prepare TEM cross-section samples of LaAlO3 single crystal subtrate with a few thousand angstroms of YBCO film deposited on the surface. So far we haven't been successful using ion beam thinning techniques.
Any help would be greatly appreciated.
Thanks in advance
Fred Pearson email: eoptics-at-mcmaster.ca
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
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I have ordered a read write cd rom to back up my digital images. The unit I ordered is backordered for another month and I am reading some reports that it is not a good unit. I have a chance to change my order for another brand which is a La Cie 4X CD recorder, from Teac, I believe. Does anyone have any experience with this make our could anyone give me some advice or comments on what they are using. Thank you very much. Gary Zajic gzajic-at-amgen.com
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RE} TEM:YBCO Sample Prep 4/29/98
I made a similar cross-section sample with YBCO film on LaAlO3. The trick is ion-milling only from the back side of the film (substrate side). To do this, you need a pair of shields mounted on both top and bottom plates of the ion mill holder, leaving an opening about 90 degree. When mounting sample, put substrate side towards opening and film side away from opening. The ion milling time is much longer than normal, but it gives great results.
Chengyu Song National Center for Electron Microscopy Lawrence Berkeley National Laboratory 510-486-6751
by orac.early.com (8.8.7/8.8.7) with SMTP id QAA17562 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 29 Apr 1998 16:59:33 -0400 (EDT) Message-ID: {004a01bd73b1$1e052060$1cd6aacc-at-rafael}
I work in a materials testing lab (minerals, steels, refractories, polymers, papers and pigment coatings) using SEM, TEM and EDS methods. We are forging into the ISO Guide 25 culture and need to be involved in proficiency tests and/or round robins. We currently report elemental composition and particle size data. I've never been involved in this type of testing before and will appreciate info and comments about the process.
Nan Laudenslager Specialty Minerals, Inc. nhl-at-early.com Phone 610-250-3094 Fax 610-250-3206
This note is in response to a recent posting in this list server quoting sections of an April 20th Business Week article addressing the lifetime of data on digital media. The author of the posting indicated that CD storage was a popular choice for the readers of the list server, so I wanted to clarify some statements in the BW article.
First off, the quoted BW comments about the lifetime of CD media address CD-ROM, not recordable CD-R. The differences between the two media are significant (see http://www.kodak.com/daiHome/techInfo/permanence1.shtml for lots of information on the permanence, care, and handling of CD-R media).
Secondly, although some CD-ROM media have shown limited life in accelerated keeping studies, studies published by 3M substantiate an estimated life of their CD-ROM media of over 50 years (William P. Murray, NMLBITS, Newsletter of the National Media Laboratory, 2(2), 4, October 1992).
Thirdly, Kodak has invested considerable effort and expense to develop a CD-R medium with excellent data life which extensive accelerated testing allows us to estimate as exceeding 100 years when properly stored (Doug Stinson et al., NMLBITS, Newsletter of the National Media Laboratory, 9(1), 1, January/February 1995).
The bottom line is that data longevity is a media quality parameter. Like anything else, some CDs (and some CD-Rs) are made to last longer than others. If data longevity is important to you, then choose a supplier that meets your needs.
Job Vacancy in the Department of Materials Science and Engineering at Lehigh University
The Microscopy Center at Lehigh University is looking for an innovative individual to develop and maintain one of the top facilities in the nation for electron microscopy. Specific responsibilities include developing microscopy techniques related to computation and developing remote operation of microscopes for teaching and research. In addition, individual will maintain instrument and computer equipment and make networking and image transfer a standard for all our instruments.
To qualify, you should have a BS in computer science, electronics or electrical engineering and at least three years related experience. Experience with electron microscopy or other scientific instruments required. Good communication and interpersonal skills, a background in laboratory and classroom environment and PC/MAC software preferred. Overtime may be required.
For consideration, please send resume before May 15, 1998 to: Dr. Alwyn Eades, Materials Science and Engineering Department, Lehigh University, 5 East Packer Avenue, Bethlehem, PA 18015. Lehigh University is committed to recruiting, retaining, and tenuring women and minorities.
Sharon L. Coe Conference Coordinator Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem, PA 18015 Phone: 610/758-5133 Fax: 610/758-4244 e-mail: slc6-at-lehigh.edu http://www.lehigh.edu/~inmatsci/Microscourses.html
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have ordered a read write cd rom to back up my digital } images. The unit I ordered is backordered for another } month and I am reading some reports that it is not a good } unit. I have a chance to change my order for another } brand which is a La Cie 4X CD recorder, from Teac, I believe. } Does anyone have any experience with this make our could } anyone give me some advice or comments on what they are } using. Thank you very much. } Gary Zajic } gzajic-at-amgen.com } }
The stability and longevity of images stored on CD-R media is a constant worry both for myself and I am sure a good number of microscopists world-wide. We have been archiving images using CD-R for five years now and have approx. 25,000 images stored at present. In the archive I have had no failures yet, but a small number of discs which I have supplied to customers have failed within a year. The CD's in question were stored in normal conditions and the only difference was that they were carried around. There was no visible damage on any of the failures. The situation at present is that we are considering installing a CD duplicator to make back-up copies of the archive. It worries me when I see lifetimes of 50-100 years mentioned in relation to CD-R media. It places a great deal of faith in dye technology which in my experience should be treated with caution. Unwittingly people reading about lifetimes like this may store important data which may be irretrievably lost.
Colin Reid Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message-----
I am an honors student, whose project is on the growth mechanism of filiform corrosion in Aluminium. I'm finishing up my literature search, and as such am covering all my bases. Is there anyone, who is doing or has done work in this field who can recommend texts papers, internet sites, videos etc that they found useful and informative.
We are currently investigating the possibility of replacing our Cambridge S-250 SEM with a new instrument. All of my experience the past 22 years as a hands-on user has been with conventional SEM's equipped with tungsten filaments but not having cryo-stages. EDS has been an extensively used option. I welcome any comments from users or vendors comparing tungsten, LaB 6, and field emission electron beam sources. Our samples are 60-70% textiles, the remainder biologicals (fungi, botanical, etc.).
Thank you in advance.
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124
Kovex Corporation announces the introduction of a new Confocal technology designed specifically for the industrial materials market. Please see our Web site at Kovexcorp.com or contact us directly at 612-486-9830/ kovex-at-spacestar.net for more details on KovexVision.
Bruce E. Batten, Ph.D. Kovex Corporation 3711 Lexington Ave. Shoreview, MN 55126 phone- 612-486-9830 fax- 612-486-9785
{HTML} Kovex Corporation announces the introduction of a new Confocal technology designed specifically for the industrial materials market. Please see our Web site at Kovexcorp.com or contact us {B} directly {/B} at 612-486-9830/ kovex-at-spacestar.net for more details on KovexVision.
} I have ordered a read write cd rom to back up my digital } images. The unit I ordered is backordered for another } month and I am reading some reports that it is not a good } unit. I have a chance to change my order for another } brand which is a La Cie 4X CD recorder, from Teac, I believe. } Does anyone have any experience with this make our could } anyone give me some advice or comments on what they are } using. Thank you very much. } Gary Zajic } gzajic-at-amgen.com }
We recently purchased a 4x12x CD-R from a different OEM (ClubMac) which also uses the Teac mechanism, and have had no problems with it. The system only a few months old, so I can't speak to long-term reliability or maintenance, but it was easy to set up and use and has yet to unsuccessfully burn a CD. We use Astarte Toast software from a PowerMac 8100/100 with an AV hard drive dedicated to CD mastering.
-Paul
Paul Voyles Loomis Laboratory of Physics University of Illinois at Urbana-Champaign 1110 W. Green St. Urbana, IL 61801 pvoyles-at-physics.uiuc.edu
I use a Yamaha CDR 4260, 2x4x6x. They make some of the most reliable drives on the market. "Smart and Friendly" uses the Yamaha drive for their product line.
} We are using a "Smart and Friendly" 2X6 CD recorder CD-R 2006 Plus and } find it ideal for backing up and saving digital images. } } --------------------------- } Miss Peta Clode } Zoology Department } LaTrobe University } Bundoora, Victoria } Australia. 3083. } } Ph (03) 9479 2177 / 2279 } Fax (03) 9479 1551 } --------------------------- } } On Wed, 29 Apr 1998, Gary H. Zajic wrote: } } } } } I have ordered a read write cd rom to back up my digital } } images. The unit I ordered is backordered for another } } month and I am reading some reports that it is not a good } } unit. I have a chance to change my order for another } } brand which is a La Cie 4X CD recorder, from Teac, I believe. } } Does anyone have any experience with this make our could } } anyone give me some advice or comments on what they are } } using. Thank you very much. } } Gary Zajic } } gzajic-at-amgen.com } } } }
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
CD-RW media are not readable on regular CD drives. Only CD-Rs can be read on regular CD drives.
Michael Bode Soft Imaging System Corp.
} ---------- } From: Peta Clode[SMTP:pclode-at-zoo.latrobe.edu.au] } Sent: Wednesday, April 29, 1998 9:16 PM } To: Gary H. Zajic } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: CD Burner } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } We are using a "Smart and Friendly" 2X6 CD recorder CD-R 2006 Plus and } find it ideal for backing up and saving digital images. } } --------------------------- } Miss Peta Clode } Zoology Department } LaTrobe University } Bundoora, Victoria } Australia. 3083. } } Ph (03) 9479 2177 / 2279 } Fax (03) 9479 1551 } --------------------------- } } On Wed, 29 Apr 1998, Gary H. Zajic wrote: } } } } ---------------------------------------------------------------------- } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------- } -. } } } } I have ordered a read write cd rom to back up my digital } } images. The unit I ordered is backordered for another } } month and I am reading some reports that it is not a good } } unit. I have a chance to change my order for another } } brand which is a La Cie 4X CD recorder, from Teac, I believe. } } Does anyone have any experience with this make our could } } anyone give me some advice or comments on what they are } } using. Thank you very much. } } Gary Zajic } } gzajic-at-amgen.com } } } } } }
Hello world, I would like any recommendations for the purchase of a new critical point dryer. We do biological materials (botanical, animal, microbiological, etc.), but not by the Gallon!! So, something nice and small and user friendly is preferable! Thanks!! Tracey Tracey M. Pepper Bessey Microscopy Facility Iowa State University Ames, IA 50011
Hi there, In our lab we are trying to accumulate all the MSDS's for the chemicals we use and put them into a 'condensed' form where if there was an emergency, the appropriate information could be easily found. The main point of this was for the clean up of spillages.
We have been looking at our MSDS for Agar 100, an epoxy resin, and it mentions in its MSDS (from the Manufacturer) that if there is a major spillage of the first 3 components (Agar, DDSA, MNA), to treat with an emulsifier then flush with water. Then if there is a major spillage of the accelerator (BDMA), totreat with a dilute mineral acid then flush with water.
We contacted the supplier to find out exactly what emulsifier to use and exactly what mineral acid to use. Even their own representative in our area didnt get a reply to his faxes (depsite replies to others!) so we figure that they arent too sure.
Does anyone know what 'emulsifier' we should have in the lab? What mineral acid should we have? How dilute? How would we clean a major spill of the resin when mixed up?! (epoxies and acclerator)
I would imagine there are other people out there in microscopy labs who try to decifer the sometimes 'vague' nature of MSDS's.
HELP!
Rich.
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
Our medical photography department is going to purchase a high end color printer- for use by the entire hospital and research facilities.
The choice seems to be between the Codonics and the Fujix Pictrography 3000. I have been hearing alot about the Codonics, but I would like to hear from some users of the Pictrography- ease, speed, resolution for EM, cost, ect.
Responding to the message of {862565F6.005E21D8.00-at-mta1.imation.com} from "Terry D. Krueger" {tdkrueger-at-imation.com} : } } Thanks for clearing up the confusion about CD-Roms. As an engineer working } in CD ROM production, I was alarmed that people might be led into thinking } that CD-ROM's were short lived media. Our guarantee is for 30 years, and } we haven't had any complaints yet. 3M doesn't make CD-Roms anymore-that } was handed off to Imation. Now Imation is handing it off to another } company (one of four prospective buyers). Some people make crappy discs } that only last 5 years, but we don't. The main problem with longevity is } the relaxing of electrical test parameters, especially bler. People should } know what the specification for Bler is before they make a decision on who } to use as their CD-ROM supplier. I am not on the Microscopy List so, could } you post this for me?
Please reply directly to the above engineer.
--
Darryl Krueger Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX.
For information on CD-R, I recommend the Adaptec site. They run a CD-R list, and maintain searchable archives, with a home page at:
http://www.adaptec.com/support/cdrlist/index.html
There has been a lot of discussion about which drives are good, and how to solve problems with some drives.
There is a new standard for CD-ROM drives, called MultiRead, which CAN read CD-RW. Older CD-ROM drives cannot. For more information, see the Adaptec site. They have an excellent glossary and other reference information.
Karen Wetmore
At 02:42 PM 29-04-98 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Bruce, I would reinstall your Cas 3.10 program. The problem is occurring on your PC and has nothing to do with the Mac's, or B.) blame the zip drive, where you a getting write errors. This can be caused by a bad scsi cable, the zip scsi card, or a bad mother board. Also, you could test this with Cas 2.5 instead of 3.10. If the problem continues it is the zip and computer mother board. You could also test this by ftping files from the PC hard drive to a central server then download. If these open without any problems, it points to the zip as the problem. I personally hate zips. They tend to be very problematic, so I have banned them from my facility. We use a large central file server, that serves as a temporary holding site for all images. Users download these over the network via ftp. Works great and serves all platforms. Our archival is done on 4.6 Gig optical disks, 600 opticals, or 640 MOs. These are far more reliable, and I have not lost any data from these in over 5 years.
Joe Goodhouse Confocal / EM Core Facility Molecular Biology Princeton University
} } Most SEMs don't have anything but steel around the chamber. Some have } } glass ports for specimen airlocks, but the HV is generally } } interlocked. I had a customer in Canada who made a Plexiglass port and } } often ran 12-18 hour EDS scans. He put a film badge inside the metal } } cover for this port and left it there for a month. He got no report of } } dosage back after turning in the badge. } } Which I for one find very worryinging. } }
It seems to me that I remember that for some reason or other, the film badges run from clear to black with increasing exposure BUT then back to clear upon extrememly high exposure - which isn't usually a problem since by the time the badges hit the re-clearing range radiation burns are clear evident and death of the wearer is soon to follow; thus, the need of the film badge (low level detection) has become moot.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
A friend (who has been asked to set this system up) asked me to post the following question:
"For under $10K, which is the best B&W Cooled, Integrating CCD Camera for the money? The camera will be attached to an Olympus BH 200. Unfortunately, I can't specify what fluorescent staining will be used but my guess is that the specimen will be neurons with GABA receptors. Thanks, Ed."
If you reply to the listserver or directly to me, I will gladly pass on the information. However, you can also respond directly to Ed at: bsgphy4-at-uconnvm.uconn.edu
Thanks in advance.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
If you are going out for a new SEM, do look into cryo stages. We don't use it often because of the time commitment but sometimes it is the only way to go. Cryo is especially useful for embyonic tissue which is so very delicate and often critical point dries poorly. Fungi also does quite well with cryo-SEM. Food scientists, Bio-medical engineers, and pharmacists have used it for such samples as starch gels and other types of gels which cannot be dehydrated without loosing integrity of the material.
Debby Sherman, Manager Microscopy Center in Agriculture Purdue University West Lafayette, IN 47907
--------------------------------------
We are currently investigating the possibility of replacing our Cambridge S-250 SEM with a new instrument. All of my experience the past 22 years as a hands-on user has been with conventional SEM's equipped with tungsten filaments but not having cryo-stages. EDS has been an extensively used option. I welcome any comments from users or vendors comparing tungsten, LaB 6, and field emission electron beam sources. Our samples are 60-70% textiles, the remainder biologicals (fungi, botanical, etc.).
Thank you in advance.
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124
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It seems to me to be important to comment that there are two ways to look at radiation issues in an electron beam instrument; i.e., 1) from an intellectual viewpoint, understanding what is taking place in the microscope, and 2) from the point of view of abiding by the various rules and regulations regarding radiation safety in force in the region/country/administration in which you are working. This thread, I'm sure, is more an intellectual discussion, but perhaps we should emphasize that: ***UNLESS YOU ARE ABSOLUTELY CERTAIN YOU KNOW WHAT YOU ARE DOING, DO NOT MAKE PHYSICAL ALTERATIONS OF ANY SORT TO YOUR ELECTRON BEAM INSTRUMENT***. The ways that x-rays can escape from a column can be quite subtle. It is very important to check, both when an instrument is new and when any alterations are made or accessories added, that it is within regulatory limits for x-ray emission. Even highly reputable manufacturers have been known to make design errors - this is especially an issue for a prototype accessory. The checks should be made in the worst-case conditions as well as normal operating conditions, and with the column deliberately misaligned (for example, x-rays may be generated by the mis-aligned beam hitting a column liner tube; poor shielding may allow them to escape, but this would not be detected if the instrument is only checked with an aligned beam).
This said, I will now continue with my main comments, which are more intellectual (at least, I hope they are!) I'm not taking issue with what has gone before - just adding my own thoughts, in no particular order.
At least half of the x-rays generated by electron interactions are Bremmstrahlung. The energy of the Bremmstrahlung x-rays extends up to the beam voltage, roughly according to the equation: I= Io.(Eo-E)/E, where I is the intensity at energy E, Eo is the incident electron energy and Io is a "constant" (which depends upon the beam energy itself, the material and thickness of the target, the beam current, etc.) In a TEM, the measured background (from a thin sample) does approximate this equation quite well, but in the SEM you also have to account for the loss of energy of the electrons as they propagate through the bulk sample, as well as the absorption of the x-rays as they leave the sample.
Characteristic x-rays, of course, have an energy threshold - that is, a minimum electron energy capable of generating them at all. Take the example of iron K x-rays. The threshold energy is a little over 7 KeV. Electrons below this energy cannot excite iron x-rays. As the electron energy is increased, the probability of exciting the x-ray increases, quite rapidly at first, eventually reaching a broad peak at something like three times the threshold energy. As the electron energy increases more, the probability of generating the x-ray falls slowly. Not shown in the way I have discussed the Bremmstrahlung above (where I only indicate the dependence of the emission on the emitted x-ray energy) is the fact that the Bremmstrahlung probability (at a given x-ray energy) also falls with increasing electron energy, rather faster than the probability of producing a characteristic x-ray. This is one of the main reasons for using high electron energies in analytical TEMs - the peak-to-background ratio for most characteristic lines improves with increasing electron energy.
Absorption of x-rays by materials is highly energy-dependent. It gets quite difficult to prevent high-energy x-rays from escaping from a microscope. By the same token, fewer of them are stopped by any biological material (for example, someone's body) they may encounter. However, those that are absorbed deposit more energy per event, and I imagine (though this is not in my field) that the energy of a particular x-ray is a rather crucial determinant in the amount of harm it can do to tissue. Also, there is more chance for the x-ray to be absorbed in some crititcal internal organ (at lower energies, they would be absorbed in tissue near the skin, presumably).
Almost any reasonable thickness of steel, brass, copper, etc. will provide shielding for 30KeV x-rays generated in an SEM. However, aluminum should not be used. A substantial fraction (I did the calculation once, and forget the exact answer) of 30Kev x-rays penetrate 3mm of Al. Even more penetrate a 3mm. quartz viewport. Similarly, a component like, for example, a thin-walled bellows is quite unsuitable as an x-ray shield for an SEM.
Interestingly, a FEG microscope generates fewer x-rays than a thermionic scope. This is because in a thermionic scaope, all of the emitted electrons are accelerated to the high voltage. A large fraction are intercepted by the first anode, where, of course, they generate x-rays. In contrast, in the FEG, the majority of the emitted electrons hit the extractor electrode with an energy of a few KeV, leaving only a small fraction to reach the full beam energy. This comment is only of minor interest, because the gun area is invariably well-shielded by the manufacturer, and tends not to be a part of the microscope that is modified later.
Sorry about the length, but hope my comments are interesting!
ZEISS EM-10A TRANSMISSION ELECTRON MICROSCOPE A 100 kV instrument with magnification of 200,000 X. Having the = following capabilities: Electron Diffraction=20 Goniometer stage and controls Television camera and monitor Both 3 1/4 X 4 1/4 sheet film and 70 mm roll film cameras Anti-contamination cold finger Nestlab water chiller Complete set of manuals including service manual and schematics
The EM-10A is located in Moscow Idaho. Contact Randal Ownbey at 208-885-2091=20 or Russ Foster at 208-885-6255=20
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.2106.6"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} ZEISS EM-10A TRANSMISSION ELECTRON MICROSCOPE {/DIV} {DIV} A 100 kV instrument with magnification of 200,000 X. Having = the=20 following capabilities: {BR} Electron Diffraction {BR} Goniometer stage and =
controls {BR} Television camera and monitor {BR} Both 3 1/4 X 4 1/4 sheet = film and=20 70 mm roll film cameras {BR} Anti-contamination cold finger {BR} Nestlab = water=20 chiller {BR} Complete set of manuals including service manual and=20 schematics {BR} {BR} The EM-10A is located in Moscow Idaho. {BR} {FONT = size=3D+1} {A=20 href=3D"mailto:surplus-at-uidaho.edu"} Contact Randal Ownbey {/A} {/FONT} at=20 208-885-2091 {BR} {FONT size=3D+1} {A = href=3D"mailto:surplus-at-uidaho.edu"} or Russ=20 Foster {/A} {/FONT} at 208-885-6255 {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D+1} {A = href=3D"http://www.dfm.uidaho.edu/surplus/Bidlist.htm"} UI=20 Surplus Bid List {/A} {/FONT} {/DIV} {DIV} {FONT size=3D+1} {DIV} {FONT size=3D+1} {A=20 href=3D"http://www.dfm.uidaho.edu/surplus/images/5044.jpg"} View Image of =
We have an Emscope SC500 coater if anyone wants it. It hasn't been used for several years, so I can't personally vouch for its integrity. Rumour has it that the only problem with it is vacuum-related....but it would at least be useful for someone as a source of spare parts, if there are any still in use.
All we ask is that you pay to ship the beast, and it is yours! If you are the least bit interested, please let me know - the boss really wants to toss this. So the offer only stands for 2 weeks, then the coater becomes scrap metal.
Luc Harmsen wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi all } } I wonder if somebody out there can help. } We are looking for a power regulator board for a Cambridge S200 SEM, part number 852594. } Leo can make one up for us, but it will take up to six months to deliver as it is an old model. } Please contact us if you can supply. } } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za
Hello,
What is the condition of your regulator board? You are rather far from me but I could probably rebuild your board, if you wish to send it to my company. I would guess about a two week turnaround and I coulld send you a quotation after evaluating the board.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. Number 499, Post Office Box 19400 Austin, Texas 78760
We are a start-up laboratory consisting of one science dropout (after 4 postdocs) who got tired of the politics and is now doing carpentry for a living, and one frustrated-by-circumstance physicist who is now a millwright. We bought an SEM in an online auction and are attempting a return to science, or at least a way to combat our midlife crises.
We want your sputter coater, and will gladly pay shipping and handling. Thank you, kind people. May the wind be always at your back, and may you find a critical point dryer to give away also.
Dear all, Does anyone know how to do purify chitinases from Pseudonocardia spp. properly? I tried to use ammonium sulfate to precipitate protein firstly,then dissolve in distilled water and dialysis against Tris -HCl . After detection chitinase by p-DAB method, we found negative result even this culture showed clear zone on chitin agar medium widely after 4 days of incubation time. Any suggestions please don't hesitate to reply me :busaya-at-kanate.su.ac.th
Free web-based email, Forever, From anywhere! http://www.mailexcite.com
Dear all, Does anyone know how to do purify chitinases from Pseudonocardia spp. properly? I tried to use ammonium sulfate to precipitate protein firstly,then dissolve in distilled water and dialysis against Tris -HCl . After detection chitinase by p-DAB method, we found negative result even this culture showed clear zone on chitin agar medium widely after 4 days of incubation time. Any suggestions please don't hesitate to reply me :busaya-at-kanate.su.ac.th
Free web-based email, Forever, From anywhere! http://www.mailexcite.com
Dear all, Does anyone know how to do purify chitinases from Pseudonocardia spp. properly? I tried to use ammonium sulfate to precipitate protein firstly,then dissolve in distilled water and dialysis against Tris -HCl . After detection chitinase by p-DAB method, we found negative result even this culture showed clear zone on chitin agar medium widely after 4 days of incubation time. Any suggestions please don't hesitate to reply me :busaya-at-kanate.su.ac.th
Free web-based email, Forever, From anywhere! http://www.mailexcite.com
Dear all, Does anyone know how to do purify chitinases from Pseudonocardia spp. properly? I tried to use ammonium sulfate to precipitate protein firstly,then dissolve in distilled water and dialysis against Tris -HCl . After detection chitinase by p-DAB method, we found negative result even this culture showed clear zone on chitin agar medium widely after 4 days of incubation time. Any suggestions please don't hesitate to reply me :busaya-at-kanate.su.ac.th
Free web-based email, Forever, From anywhere! http://www.mailexcite.com
Dear all, Does anyone know how to do purify chitinases from Pseudonocardia spp. properly? I tried to use ammonium sulfate to precipitate protein firstly,then dissolve in distilled water and dialysis against Tris -HCl . After detection chitinase by p-DAB method, we found negative result even this culture showed clear zone on chitin agar medium widely after 4 days of incubation time. Any suggestions please don't hesitate to reply me :busaya-at-kanate.su.ac.th
Free web-based email, Forever, From anywhere! http://www.mailexcite.com
The Chesapeake Society for Microscopy, a local affiliate society of the Microscopy Society of America, will hold it's annual Poster Meeting at the Uniformed Services Univ for the Health Sciences (USUHS) in Bethesda, MD, on Tuesday, May 19, from 6-9 PM. Program info is available at the website:
http://162.129.38.210:8001/csm_home.html
For poster entry registration and details contact: Brian Andrews sba-at-helix.nih.gov
This is a list of some of the responses I received to my query regarding opinions on components of image analysis systems. Thanks to all who responded, you have been helpful.
MY REQUEST: Re: Image Analysis Systems (microscopes, CCD cameras, and software). I am interested in hearing anyone's views or preferences as to the components/manufacturers. I have lists of microscope and camera manufacturers, but would appreciate comments from your experience with the use, servicing, etc, of such equipment (and recommended software).
RESPONSES: These are summaries of the responses that I still have.
1) I would like to point your interest to our website:
http://www.soft-imaging.de
where you may find some useful nformation on our products. If you are interested, I would be pleased to send some more printed information to you. If you would like to receive it, please fill out the form on our website.
Best regards, Christian Mellen Soft Imaging System GmbH
2)
Working out what the best image analysis system is is quite a can of worms. From my experience the best thing to do is to fit the system to the type of images you need to look at. The mistake many people make when approaching image analysis is to think the computer will solve all their problems. If you cannot easily distinguish objects the computer probably will not either, if you can distingush them then the computer may still have problems. Often the hard work in any image analysis problem is getting the image into a state so that measurments can be easily carried out. The choise of a system often depends on how much effort you want to put into developing automatic routines vs semi-automatic/interactive operation - the latter would generally be cheaper both in cost and development time.
I would hesitate to recommend any single system. I have been reletively happy with our set up (now 6 years old) of monochrome and 3 chip colour video cameras, Matrox imaging card and Optimas software. One suggestion in making a purchase is to have a clear set of parameters the system must be able to meet and a return clause if it fails. Our original (very expensive) software purchase failed in a number of aspects and was replaced by the cheaper Optimas alternative.
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
3)
Greetings from DVC Company... Yes we are a monochrome CCD digital and analog camera manufacturer and supply the framegrabbers and boards......... See all info below and on our web http://members.aol.com/dvcco Let us work with you and with Acrobat reader you can download our files off the web for review and printout...manual.pdf and datasht.pdf alternate site for DVC10 series cameras only www.optics.org/dvc For reference you can chat with : David Beaglehole Beaglehole's Ellipsometry Site Physics Department http://www.vuw.ac.nz/~db/bes/bes.html Laby Building, Victoria University of Wellington, POBox 600, Wellington, New Zealand Tel 64 4 471 5347, Fax 64 4 495 5237
DVC Company is a US Camera Manufacturer and systems house for frame grabbers and software, that offers: All units are monochrome outputs. Color can be had with our Tuneable RGB sequencial filter / non real time, if needed.
DVC does the custom digital RS-422 cables for various frame grabber boards.
The DVC-10 is a monochrome 10 bit real time analog and digital CCD Area array cameras that are very sensitive with signal to noise rating of } 62dB with capability of 30dB of gain if needed and simultaneous digital RS-422 and RS-170 outputs. We specialize in microscopy and will work with your people. DVC is also a complete systems house suppling 15 lines of frame grabber boards including Matrox !!! boards, and software. We cover PCI bus for PC and MAC, SUN and also SGI Web sites have downloadable files via Acrobat 3.0 reader / 3 files.. and complete camera manual. We are on any web browser via the sites listed below.
http://members.aol.com/dvcco This web site has been updated with new DVC-1300 info and pricing...Check this one first.
Feel free to call us with any questions Please pass our web site on......your e-mail system if you choose.
Regards,
Richard Klotsche DVC Company / San Diego, CA 619-444-8300 619-444-8321-fax
4)
Dear Listservers,
I am going to use UMAX Powerlook III scanner with transparency attachment to digitise 3 1/4 x 4" (8.3 x 10.2 m) TEM negatives. Does anyone know of a supplier of a film holder that can be used with the scanner? I contacted UMAX 10 days ago, but they have not responded so far, perhaps they do not have one to accommodate this size. Would the scanner recognise a home made film holder?
Thanks, Alex ______________________________ Alexander Titkov Millennium Inorganic Chemicals PO Box 245 Bunbury WA 6231 Australia Ph: (08) 97 808 505 FAX: (08) 97 808 500 E-mail: atitkov-at-micl.com.au
We are often used by clients, either in the run up to a purchase, or for the total task in an advisory capacity. Having worked for several microscope marketing organisations as demonstrators and sales specialist much of the "sales and demonstration hype" we see and overcome. In this way the purchase procedure is not muddied through the tricks we all know the clients fall for and the sales team thrive, often rely, upon.
Important points are set out below and expanded in the attached.
It is all to easy to follow the "we know how brand X work lets buy their new one" syndrome when considering a new electron microscope, unfortunate= ly you may not always best serve your unit's clients with this approach! =
Remember the SEM chamber geometry sets the information that you see, different shaped chambers will image the same specimens in a different wa= y. There will be one instrument that will handle your specimens better than=
any other, the clever trick is to find it!
Bring together all the people who are likely to use the instrument and dr= aw up a list of your requirements. Grade them in a 1 to 10 "priority list".= =
Obtain ALL the brochures and sift through the ones that are nowhere near your requirements. Do not stick to the ones you know that you can afford= , look at those you feel may be a little out of reach too.
Select a maximum of four specimens from the users, specimens with a wide range of different requirements. Go and see all of the manufacturers on your short list. the demo is the most important day remember:- 1. You are buying the microscope, not the demonstrator (?) help them= =2E 2. Stick rigidly to looking at your specimens 3. Follow a precise procedure that you will use for each demonstrati= on 4. Time the production of your results from start to finish of each specimen 5. If you have done all your work let the demonstrator show you some=
of the other areas of the instruments that may be of interest.
After the demo collect all the results and have the group of users look a= t all of them and grade the results. If you do not have an almost identica= l range of results in truth the demonstration was a waste of time! Remembe= r, if you buy an instrument that fails to perform as good as the demonstration, a purchase set against a good comparative series of photographs is easier to use as a standard than gut feelings? Use the photographs, and if the installed instrument is poor get it fixed or changed, use just gut feelings to select and you are dead! EM Quality starts by getting the best instrument for the task and by making sure tha= t it performs to specification; insist you have 100% satisfaction.
Select the instruments that come out on top with relation to the results,=
from now on only consider the first two instruments. Talk to the manufacturers and see if you can bargain with them to obtain more for you= r money. The best bargain you can obtain, balanced against the results it achieved, should be your purchase. Sounds simple but we will expand on t= he important areas often missed by prospective buyers.
Remember more information is attached.
Steve Chapman Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Buying A New Instrument? =0D How do you start?=0D =96 Collect ALL the brochures AND Prices=0D =96 Form a view of the new facilities available=0D =96 If you do not understand them ASK the salesman=0D Check ALL possible users desired facilities?=0D =96 Those who use the instruments now?=0D =96 Those who may use them in the future if you buy certain accessories?=0D=
Formulate a purchase specification=0D =96 Price Range - Also look at one level HIGH=0D =96 Set essential Instrument specifications=0D =96 Set desired instrument specifications=0D Remember a consultant may be able to help-there approach covers=0D =96 Interdepartmental feuds that could spoil the case=0D =96 Asking the appropriate questions in a different, less pointed way, ma= y bring different conclusions=0D =96 Their far greater knowledge of the subject over a wider base than mos= t scientists will bring out points which others may miss=0D =96 After talking to all interested parties features will be given a desi= rability factor=0D =96 "Irrelevant" features will be excluded and punch features highly rate= d =0D =96 Aim at one instrument higher than the budget - he will know "the E.M.= business"=0D How Do YOU Handle a Demonstration?=0D How NOT to do it!=0D =96 Do NOT -Select YOUR favourite specimens=0D =96 Do NOT - Take enough specimens to keep you going for 6 hours or more=0D=
=96 DO NOT -Try out some totally new techniques=0D =96 Particularly those you have NOT ever looked at before=0D =96 DO NOT FALL INTO THE TRAP - Of not telling the demonstrator in advanc= e what you intend to do, if you do then do not change your mind on the da= y=0D =96 DO NOT FALL ONTO THE TRAP - You act as if the demonstrator is the ene= my=0D =96 DO NOT FALL INTO THE TRAP - He has to find out so do not help him at = all=0D =96 DO NOT FALL INTO THE TRAP - See if he is clever enough to find soluti= ons in one day to the problems that took you 5 years to overcome=0D =96 DO NOT -Try to develop a new research project during the day, use the= ir film its cheaper=0D =96 DO NOT FALL INTO THE TRAP - Do write notes, you will not remember the= detail of the demonstration.=0D How Will the Demonstrator Handle YOU?=0D Do NOT let him!=0D =96 Talk all day about the wonderful instrument features - you came for a= demonstration!=0D =96 Dominate the demonstration - it is YOUR demo=0D =96 Demonstrate how wonderful the alignment system is=0D =96 Show you the fantastic number of stage memory points you can remembe= r=0D =96 Show you HIS favourite specimen=0D =96 Play with his favourite gimmick on the instrument=0D =96 Take you out of the demonstration room when you have just given him a= new specimen=0D =96 Fill you with food and drink during a 2 hour lunch break=0D Before the Demo=0D =96 Select three or four specimens that are important to your laboratory=0D=
=96 Specimens that will test an instrument be it low or high magnificatio= n, no matter=0D =96 Develop a demonstration criteria - a specification for each specimen=0D=
=96 Provide each company with an exact demonstration programme=0D During the demo=0D =96 Time the demonstrator from start to finish with each specimen e.g. t= ake pictures at 2,000X, 4,000X, 16000X, and 32,000X=0D =96 Encourage the demonstrator not only to use your operating specificati= on for a task but also to try those he feels are best for this instrument= in these circumstances=0D =96 Prevent the demonstrator doing what he wants if this has no bearing u= pon your applications=0D After the Demo=0D =96 Layout the results=0D =96 Compare instrument with instrument on each specimen relating image qu= ality to time taken to produce the results.=0D You have to assumer that ALL the demonstrators were of equal caliber, do = not use gut feeling!=0D =96 Produce a short list =0D =96 Compare the short list against the "desirability" assessment=0D Research=0D =96 Look at the organisations who would provide the instruments=0D =96 Their service record in a number of laboratories nation wide=0D =96 Their service record in a number of laboratories in your area=0D =96 The reputation of the production company=0D The Purchase=0D =96 Decide upon the instrument you need and its reserve.=0D =96 Decide upon the specifications required.=0D =96 Approach the appropriate manufacturers, get the deals in writing incl= uding ALL the odds and ends mentioned.=0D =96 Haggle, get THE FINAL deal in writing including ALL the odds and ends= mentioned.=0D =0D Published by Protrain, Chinnor, Oxford, England=0D Tel & Fax 44 (0)1844 353161=0D E-Mail protrain-at-compuserve.com=0D Web http:/ourworld.compuserve.com/homepages/protrain=0D
} I am going to use UMAX Powerlook III scanner with transparency attachment } to digitise 3 1/4 x 4" (8.3 x 10.2 m) TEM negatives. Does anyone know of a } supplier of a film holder that can be used with the scanner? I contacted } UMAX 10 days ago, but they have not responded so far, perhaps they do not } have one to accommodate this size. Would the scanner recognise a home made } film holder?
Does the holder have any effect on resolution? I doubt that it does more than just hold the transparency in place. In which case, just cut one out of a sheet of plastic. I don't usually bother with a holder at all.
MAMAS (Mid-Atlantic Microbeam Analysis Society) and the Surface and Microanalysis Science Division,NIST Meeting at the National Institute of Standards and Technology Gaithersburg, MD on Thursday, May 14, 1997 10:30 am- 3:00 PM Lecture Room C, Administration Bldg.
10:30am Coffee and Doughnuts
10:45am S. Brian Andrews, National Institutes of Health "Recent Advances in Biological Electron Probe Microanalysis"
12 noon Lunch
1:15pm Joe Michael, Sandia National Laboratory "Phase Identification using Electron Backscatter Diffraction in the SEM: an Alternative to Electron Diffration in the TEM"
The Minnesota Microscopy Society's annual Spring Symposium is tommorrow, Tuesday May 5th, at the Sheraton Midway hotel St. Paul (I94 and Hamline)
The topic is "Microscopy of Biomaterials"
Agenda
8:30 Registration
9:00 Microscopy of BioMaterials: An Overview Patrick Parks, 3M BioMaterials Technology Center
9:45 Aligned Artificial Collagen Systems Ted Tower, Dept. Chemical Engineering & Materials Science, University of Minnesota
10:30 Break / Vendor Displays
11:15 The Application of Correlative Microscopy to the Study of Biological - Biomaterial Interactions Ralph Albrecht University of Wisconsin, Madison
12:00 Lunch
1:15 Structure of Teeth Professor W. Douglas Center for Biomaterials and Biomechanics, University of Minnesota
2:00 Break / Vendor Displays
2:45 Cellular Performance of Biomaterials: A Macroscopic and Microscopic Assessment Maura Donovan, Medtronic, Inc.
3:00 Vendor Displays
Symposium Fee: $25.00 current regular MMS members 97/98 (dues paid since 9/1/97), $35 non-member (includes regular membership), $10.00 student members 97/98 (dues paid since 9/1/97) $15.00 non-member students (includes student membership). Fees payable at the door, but registration preferred to Mike Coscio at (612) 569-1331, E-mail: mike.coscio-at-medtronic.com
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
I need to add a digital camera to our light microscope: either a Polaroid PDC 2000 (1600 x 1200) or a Kodak DC120 (1280 x 960) are being considered. Cost: $1,500 - 700, respectively. Anyone have any experience with these attached to LM's? Suggestions? Budget restriction: $2,000. Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Hello, We are running out of lab (and storage) space, and thus must let the following items go.
Durst Laborator S-45 EM enlarger for 4X5 or smaller negatives and plates. It has: Durst Laborator 138 condenser lens: 1 Latico 85 1 Latico 110 2 Latico 130 1 Latico 160 1 Latico 200 3 Latico 240
Final lens of 80, 105, 135 mm Negative holders- 2 Nega 138 (5X7), 1- 35mm assembly This is a point source set-up, it used to be our main system until we bought a new one. It is in working condition.
Next A Kevex Unispec system 7000 (EDS). It has a 77 main frame, 4800 ratemeter, 5130 EDC, and 4505P pulse processor. I had hoped to use the electronics in conjuction with some 4pi products to support a detector, but demand doesn't justify the budgetary outlay. The electronics were functional when it was placed in storage, but the display was having difficulties (bad yoke?).
Interested parties should contact myself or Kenneth Moore (kenneth-moore-at-uiowa.edu). -- Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
Kovex Corporation, a rapidly growing high-tech manufacturer of surface inspection equipment specializing in the development of state of the art microscopy techniques is looking for an Optical Systems Scientist. If you would like to know more about this position please check out our web page at www.kovexcorp.com
Winnie Westbrook wrote: } } Hi There, } Looking for a good technique for making a pellet of buffy coat from } normal blood. Would appreciate any help. Thanks. } } Winnie To Everyone Who Replied, Thanks for the info concerning the buffy coat collection technique. My difficulty had to do with the amount of sample I was given. It sure makes it easier with more blood sample.
As previously announced, MMMS will hold a Materials Science meeting on = May 22, 1998, at the University of Illinois at Chicago. The speakers and t= heir titles are:
8:45 Dr. Nigel Browning UIC Dept. Physics Introduction 9:00 Prof. Manfred Ruehle MPI-Stuttgart "Interface Science - More Information about Less" 9:30 Dr. Stephen Pennycook Oak Ridge Nat. Lab. "A Combined Experimental=
and Theoretical Approach to Atomic Scale Characterization of Interfa= ces"
10:00 - 10-30 COFFEE BREAK
10:30 Prof. Laurie Marks Northwestern Univ. "Who Needs Images? Solving Structures to 1 Angstrom Resolution Using Electron Diffraction" 11:00 Prof. Vinayak Dravid Northwestern Univ. "Dynamics of Electrostati= c Potential Barriers at Interfaces" 11:30 Prof. Ian Robertson UI-Champaign/Urbana "Design and Application o= f a Controlled Environment Transmission Electron Microscope"
12:00 - 1:30 LUNCH/POSTERS
1:30 Prof. John Silcox Cornell Univ. "Electron Spectrosocpy at the Atom= ic Scale" 2:00 Prof. Chas. Lyman Lehigh Univ. "Microanalysis of TEM Specimens: I= s 300kV Necessary?" 2:30 Prof. Ondrej Krivanek U. Washington "Aberration Correction in the = STEM"
3:00 COFFEE
3:30 Prof. Pedro Montano UIC/Argonne "X-Ray Resonant Reflectivity: A To= ol to Characterize Interfaces" 4:00 Prof. Tom Kelly U W Madison "3D Atomic Scale Analysis with a Loca= l Electron Atom Probe"
4:30 TOUR OF MICROSCOPE FACILITIES
For further information and travel instructions, contact jane.a.fagerland-at-abbott.com =
A little off topic, but I would like to know if any of you out there have experience doing 3-D reconstruction from cryosectioned tissue. In the literature, I've read articles on tissue distortion from cutting paraffin embedded tissue and mention of potential problems using internal fiducials.
However, my preliminary results using cryosectioned tissue seem pretty good (goal: to quantify optical density sections in relation to 3-D reconstructed anatomical structures).
Details: 20 micron thick rat brain tissue sections with embedded fiducial points, that is, I use small needles to penetrate the whole tissue using a stereotactic grid prior to embedding into OCT and sectioning, then use the holes as reference to register the digitized sections without warping the digital image.
Just wondering what others might have found. Thanks,
Brian Tryon MD/PhD Student Dept. Neurobiology & Anatomy Allegheny University of the Health Sciences 3200 Henry Avenue Philadelphia, PA 19129 USA
Currently, there are three systems utilizing or adapting midrange digital camera technologies to the microscope. If anyone wishes to add to this list, please do so. Kodak uses a stock version of the DC120 camera($700) for their MDS120 system ($2100) Polaroid adapted their PDC2000 ($1500) camera to create the DMC2000 ($5995). While you are hunting in this range of price and capability you should also consider the Pixera Professional ($1200). A standard c-mount lens turns this unit into a digital camera that must remain tethered to a computer. Last November in a similar thread, I gave a side by side comparison of these systems. Search the archives under "digital camera" or send me an email and I will send a summary.
I think you'd get a lot of argument against using any of these systems for critical imaging applications. See the current thread on the confocal listserv for the next step up in digital imaging from these systems. However, if any of these satisfy your needs they are certainly much more affordable.
I have been using the Kodak MDS120 system for a few months. I chose it for two reasons. First, they used the stock camera which can be easily unmounted and used for other tasks around the lab. It is a great digital camera. The second reason is that I needed the microscopy imaging system for fluorescence work. It is the only system which doesn't immediately warn against using the system for low light applications. In fact, I was delighted with the sensitivity of the camera for this application. General microbiological techniques such as AO or Dapi direct counts are well within its capabilities. I have also used for much lower intensity techniques such as DFA and even rRNA targeted oligo probes.
I remain interested in the Pixera system. For the price, it is certainly worth checking out. As far as the DMC2000 goes, I'll say the same thing I did last time. If I had that much to spend, I could probably rationalize and afford to leap to that next level of digital imaging capability.
Usual Disclaimer: I have a financial interest in getting out of Grad school but not in any of the aforesaid vendors.
Kevin Brent Smith University of Louisville Biology Dept. Louisville, KY 40292 Phone: (502) 852-6773 Fax: (502) 852-0725
I am looking for information on triple-labelling techniques for fluorescent secondary antibodies (including nuclear, cellular and extracellular matrix). In particular, procedures which use AMCA, CY2, CY3 and CY5 and what conditions favours the use of combinations of these markers.
I am looking for information on triple-labelling techniques for fluorescent secondary antibodies (including nuclear, cellular and extracellular matrix). In particular, procedures which use AMCA, CY2, CY3 and CY5 and what conditions favours the use of combinations of these markers.
Dear all, =20 I have used tilt correction in the past but am stuck on one specific=20 application. I have a tilted sample (70 degrees) and want to image the equiaxed=20 grains clearly visible at 0 degree tilt.... At 70 degrees, these appear elongated and checking the box "tilt=20 correction" on the Leo 435 (also specifying the tilt angle) does not=20 make a difference... Isn't it working because the image i want consists of equiaxed grains=20 and haven't a square shape? =20 Thanks =20 F.
Are you looking at (etched) topography (SE) or using "channeling contrast" (BSE)? If the latter, the actual contrast for each grain will change (with the incident beam angle). Should this be the case, no amount of correction can help...
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Dear all,
I have used tilt correction in the past but am stuck on one specific application. I have a tilted sample (70 degrees) and want to image the equiaxed grains clearly visible at 0 degree tilt.... At 70 degrees, these appear elongated and checking the box "tilt correction" on the Leo 435 (also specifying the tilt angle) does not make a difference... Isn't it working because the image i want consists of equiaxed grains and haven't a square shape?
I have just finished up with a group of freshman undergraduates with an introductory SEM course. We are trying a new idea by posting their projects on the web. If any of you are interested in reviewing these projects please go to:
There is a form block for submission of your comments at the end of each project.
Thanks!
Brian
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax) "Be well, do good work, and keep in touch"
Someone here wants to pick up plant root cryosections onto Gallium coated silicon windows. He chose Gallium to hold the sections down and for its conductivity for SIMS and SEM. The sections will be Au/Pd coated. Not being familiar myself with Gallium metal, does anyone know if the low melting point (29 degrees C) will cause a contamination problem under the SEM beam? Thanks Wallace Ambrose Dental Research Center University of North Carolina Chapel Hill, NC
A.O. 100x/1.25 oil Plan Acro cat # 1024, Needed. Any sources out there? Please email price and availability or repair costs to tosborn-at-csubak.edu. Thanks Tom
My apologies to the list, I just forgot to put in my department (molbio)
The Correction is below.
To all interested microscopists in Confocal and Biological EM, I am pleased to announce our new site on the web. Point your browser at http://www.molbio.princeton.edu/confocal/CF-EM-HOME.html to see what we do, and what we do it with. Much more to come in the near future.
Regards,
Joe Goodhouse Confocal / EM Core Facility Department of Molecular Biology Princeton University
What matters is not the melting point, but the vapor pressure. Surprisingly, the vapor pressure of most materials does not change discontinuously upon melting. The vapor pressure of gallium is reasonably low at near room temperature (the CRC, 60th ed., lists it as 1 mm Hg at 1350 degrees C). A more precise value for the vapor pressure at room temperature can be obtained from the text "A User's Guide to Vacuum Technology" by John O'Hanlon (Wiley, 1980). A good text on vacuum practices, like O'Hanlon's, may have more to say about gallium. Frankly, I don't see a problem for a short term experiment with a coated sample. I suppose it depends on your tolerance for contamination with a hazardous heavy metal.
********************************************************* Jeffrey A. Fortner, Ph.D. Chemical Technology Division Argonne National Laboratory 9700 S. Cass Avenue Argonne, IL 60439-4837
} ---------- } From: Ambrose, Wallace } Sent: Tuesday, May 5, 1998 1:17 PM } To: 'microscopy-at-sparc5.microscopy.com' } Subject: Gallium in the SEM } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Someone here wants to pick up plant root } cryosections onto Gallium coated silicon windows. He chose Gallium to } hold the sections down and for its conductivity for SIMS and SEM. The } sections will be Au/Pd coated. Not being familiar myself with Gallium } metal, does anyone know if the low melting point (29 degrees C) will } cause a contamination problem under the SEM beam? } Thanks } Wallace Ambrose } Dental Research Center } University of North Carolina } Chapel Hill, NC }
Gallium is one of those wonderful materials that breaks with intuition. Although the material has low melting point, the vapor pressure is extremely low. The vapor pressure versus temperature curve is between manganese and beryllium,
We have developed an honesty problem with the users of our Kodak 8650 Dye sub printer. The media is costly, we need to know who is using the printer and how much media they have consumed. The computer is already on NT and everyone has a password, but the 8650 is not networked {it takes around 5 minutes to print a 20meg image besides clogging up the server} and we use Photoshop 4.01 for all of our imaging.
What I need to know is if there is a mechanism in photoshop that can monitor the export usage, we have not found it if it is there; is there a device that we can install on or in the printer that can number the images used per person...or be creative like the venerated Dr. Mark Farmer from the University of Georgia who suggested the "Jersey Method" which he claims to have developed in the mid 1980s.
Any help would be appreciated.
Funny colors at sunset, strange smells, dreary, damp, dank
John Grazul Rutgers University Electron Imaging Facility
Regarding tilt correction, here is something general which applies whenever it is used.
In SEM, tilt correction is applied to the 2-dimensional picture of a 3-dimensional object. If, for example, you are imaging small spheres sitting on a plane surface, then however the specimen is tilted, without correction these would give circular outlines. With tilt correction these will be drawn out into ellipses, and the spheres will appear as if they ellipsoids (Rugby or American football).
Cubical grains will also suffer in a similar way.
Mathematically, this is quite similar to what happens in pictures taken with an ordinary wide-angle lens. If you took a group of people each holding a spherical football, the effect would show up at the edges of the picture. In this respect a fish-eye lens introduces less distortion.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Some years ago, some standard asbestos samples were developed that became known as the UICC "standards". It is a long story, but at some point in time, we ended up with either some or all of the remaining material from this well characterized set of samples.
The problem is this: We have run out of the crocidolite standard and people keep asking us for it. Does anyone out there have any of the UICC crocidolite material sitting around that they would be willing to part with, sell, trade, or whatever?
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I'm just curious as to the policies followed by multi-user imaging and microscopy facilities regarding establishment of protocols for specimen preparation and analysis. Specifically, do people follow the idea that the researcher/client should establish protocols by literature searches and experimentation and then present these protocols to the lab to be executed? Or is it expected that EM facility personnel, being the microscopy experts, will establish protocols, follow them and present the results and written protocols to the researcher/client on an independent basis? Or something in between?
We sometimes see situations where specimens are brought in for processing, and when the client is questioned as to how they want them processed, we're told "I don't know. Do it like you did it before.", even though different lab personnel were involved, records are mislaid or nonexistent (I know, I know...) etc. In other words, it appears that work is being done and results published when the primary researchers aren't aware of the materials and methods used in the process, or whether the techniques are optimal for their particular needs. These things are sometimes left to the complete discretion of lab technicians, with all the varied backgrounds lab technicians have.
Now remember, I'm discussing multi-user service facilities, not specialized labs dedicated to a single area of research. We're doing a rather radical reorganization of our facility at the time, and I'm very interested in other's thoughts on this. It will aid in setting some policy guidelines.
Thanks. If anyone wishes, I'll be happy to post a summary of the replies.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
I have a major Problem with our new HPD-CPx software controlling the Hamamatsu C4742-95 camera. When storing aquired single images using the SAVE AS... command the program crashes. This problem occurs unregularly but relatively often. Unfortunately I cant reproduce the conditions under which the failure occurs. HPD-CPx is running under Win NT 4.0 here.
Has someone made the same experience and can perhaps give me some advice how to fix this problem? Hamamatsu says it occurs only on my computer, so they assume a hardware problem (but which one???).
It is tough to have A policy. In general most samples that come in can be processed by one of a few routine techniques. From experience we already know how to adapt SOPs to a different situation, i.e., plants may need to be handled slightly differently from animals etc. In most cases our users do not have enough background to make decisions about protocols and rely on us to make the best choice. When we get a project that we know from experience, or just intuitively feel, might need special processing we will ask the investigator to provide us with a successful protocol from the literature on that sample or one related to it. We always at least get a lead to the literature if not a specific procedure. This is especially important if they want immunolabelling.
If it takes several trials to get a successful outcome, the users are billed accordingly and are warned in advance that this might occur. If we screw up, we don't charge them for repeats.
No matter what the sample, we encourage the user to talk to us before bringing the sample so that we can head off problems in advance. We may even be able to help them with their experimental design in order to optimize the results. People who show up unannounced on on short notice are warned that they are putting us at a disadvantage and that may be reflected in the outcome. We also politely give them hell for being so presumptuous to think that we operate like McDonalds drive up window.
When technical staff needs help in making this decision, that will consult with the Scientific Director of the lab. I would hope that such a person exists for your facility if it isn't you. We have also been know to get on the phone and consult with whomever we think might be helpful on a sticky project.
Hope this has been informative.
I should that we do about 500 samples a year for about 100 different investigators, almost all bringing biological samples.
Greg Erdos
} } } } } } } } } } } } } } } } } } } } } } } .
} Hi, } } I'm just curious as to the policies followed by multi-user imaging and } microscopy facilities regarding establishment of protocols for specimen } { { {snip} } } } } ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Assistant Director, The Biotechnology Program PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Dear Randy, } } I'm just curious as to the policies followed by multi-user imaging and } microscopy facilities regarding establishment of protocols for specimen } preparation and analysis. Specifically, do people follow the idea that the } researcher/client should establish protocols by literature searches and } experimentation and then present these protocols to the lab to be executed? } Or is it expected that EM facility personnel, being the microscopy } experts, will establish protocols, follow them and present the results and } written protocols to the researcher/client on an independent basis? Or } something in between? } We consult with prospective users and suggest protocols. The users can then send us grids which we examine and evaluate, and we send represen- tative pictures back. When the user is satisfied that the features of in- terest are clear, that is the protocol used. We want everyone who comes to our facility--a NIH-funded biotechnological resource--to have productive visits, so we take pains that their specimens be properly prepared for the HVEM, IVEM or other instruments at our site. Yours, Bill Tivol
} Imageers, } } We have developed an honesty problem with the users of our Kodak 8650 } Dye sub printer. The media is costly, we need to know who is using } the printer and how much media they have consumed. The computer is } already on NT and everyone has a password, but the 8650 is not } networked {it takes around 5 minutes to print a 20meg image besides } clogging up the server} and we use Photoshop 4.01 for all of our } imaging. } } What I need to know is if there is a mechanism in photoshop that can } monitor the export usage, we have not found it if it is there; is } there a device that we can install on or in the printer that can } number the images used per person...or be creative like the } venerated Dr. Mark Farmer from the University of Georgia who } suggested the "Jersey Method" which he claims to have developed in } the mid 1980s.
We have a similar setup but ours is networked to and from one NT Workstation. That is, the workstation owns the printer. We do not have a problem with the server because TCP/IP routes it directly back to the workstation; it never passes thru a server and with the network card you can use PS or raster mode. No print takes more than 70 seconds once it is sent to the printer. I have never seen anything in PhotoShop to track prints. When you say everyone has a password, do you mean they each have a password or do you mean they all use the same password? If you aren't using user profiles you can't do much. If you are using profiles you will need audit software like W3 (http://www.winwhatwhere.com).
NT can track printing by individual users if you are using user profiles. But that is for printing, not exporting. It is done like this: In User Manager/Policies/Audit select "File and Object Access". In your Printer control panel select the printer, then right click and select Properties/Security/Auditing. Then you must add your users (or their group) into the "Name:" window. Next, select the events to audit. That would be Print, Success and Fail. Save all this and then when the Print command is used it will be recorded in your System Log which you view with your Event Viewer.
Good luck.
Rick A. Harris, Director Microscopy and Image Analysis Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax raharris-at-ucdavis.edu
I run a multi-user facility at Emory University. Our policy is to follow the protocols for all general samples which we have written into a lab manual . Special protocols are developed and used only when the routine is not adequate for the researcher's sample. No lab personnel are permitted to modify protocols without direct supervision by the researcher requiring changes ( and only then by my permission).
Your observation that the materials and methods are seldom known by the remote user is quite true. To eliminate this problem, all our outside users are provided with a written copy of the protocol used to prepare their samples. Any specific unusual observations noted or steps taken during the preparation are written on the forms as the procedures are completed. This may sound like additional paperwork, and it is, but we have eliminated many arguments when samples do not turnout the way a researcher had envisioned.
I hold to the premise that lab protocols are static for the majority of sample preparation needs, therefore, follow a fixed-written routine everytime. Our consistancy of results is a testament to the truth of this premise.
We usually take the middle road: if it is a sample we have prepared before we try to follow previous procedures. However, if it is something new, I often ask the investigator (faculty or student) to do the legwork in finding previous papers with EM protocols, especially those showing the type of information they are after. This is important because it may not be clear to me whether the project is feasible at all (as you know, non-EM people sometimes are wildly optimistic about what we can detect and what we can't). I have had varying success with this approach, but it usually separates the customers who are serious about getting good results, and are willing to put in some time, from those who are just "fishing".
As for record keeping, we do keep records, but inevitably some details end up stored in the head of the person who does the work. I am sometimes horrified to find out months later that the study has been reviewed and published with a Materials and Methods section that bears only passing resemblance to what was actually done. A surprising number of people never bother to ask us to review what they have written even though they have little understanding of the techniques. The same people (fortunately!?) usually don't bother to put us in the acknowledgements, either.
Marie
} Hi, } } I'm just curious as to the policies followed by multi-user imaging and } microscopy facilities regarding establishment of protocols for specimen } preparation and analysis. . .
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
A TN 5500 EDS system is available for sale as replacement for existing system parts. The system was operable before it was dismantled last week. The system does not include the detector. Also, the printer gives some trouble if it is not frequently serviced and cleaned. The system is supposed to be complete with imaging, and analysis options. If you have interest please contact me or send me email. will sell for best offer.
thank you
== Said A. Mansour == Purdue University == School of Materials Engineering == 1289 MSEE Bldg. == W. Lafayette, IN 47907-1289 == # (765) 494-6405 Fax (765) 494-1204
I concur with Greg Erdos that we try to get the investigator to talk to us before the experiment then determine if an SOP will fit. If not then I ask the investigator if they have done a literature search. If there is a technique for their project I determine if we have the capabilities to perform it or if it realy is just a "conveniance" modification that the paper used and one of ours will indeed work. Ihave on Computer the different techniques we perform so when it comes to manuscript time I just print out a copy and any modifications for the investigater. Like Greg we get the occasional person who thinks EM is a few hour job and says here"s the tissue. Something else that I'd like to add or know is how many labs break up their services and subsequent charges into say proceesing, thicks, thins, scope time (by EM staff or investigator), and photography, or do they charge as a lump project? We break it all down and even alow the investigater to perform what ever steps they think they can. I'd like to see a survey of prices for these things too.
Rick Vaughn
PS Thanks to all the advice on stereo pairs. Hopefully you'll see them at the 99 Portland meeting.
I can't help you with your honesty problem and I can't help you with your network problem, but I can suggest a method that worked for me at Wright Patterson. We had a Kodak 8650 at Wright Patterson and there were both Mac and PC users. I collected and processed images on both platforms. It was not on the network, but was dedicated to one computer that was on the network. Other dye sub printers that were on the network had problems with usage/misusage and a lot of Mac users not knowing what printer they were printing to.
I would suggest that you take the printer off-net and put it on one computer and then have users come to the machine. That solves the network clogging problem and the waste due to accidental print jobs. Users could also buy and use their own paper. That solves the honesty problem. The computer that you use would have to have the software that you use to print. Incidentally, I typically process my images in Photoshop and save them in TIF mode and print them in Powerpoint. Regardless, Powerpoint and Photoshop are both programs that are very similar and compatible across platforms. With Powerpoint, you can easily arrange different prints and print multiple copies. I could see no difference in quality of the prints when Powerpoint and Photoshop were used. I would suggest that you use a PC running Windows and use Conversions Plus so that it can recognize and use both Mac and PC formatted disks. (It is best to use PC formatted disks and then quick format them to Mac. This method preserves the long format names on both platforms.)
Just my two cents.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
We have developed an honesty problem with the users of our Kodak 8650 Dye sub printer. The media is costly, we need to know who is using the printer and how much media they have consumed. The computer is already on NT and everyone has a password, but the 8650 is not networked {it takes around 5 minutes to print a 20meg image besides clogging up the server} and we use Photoshop 4.01 for all of our imaging.
What I need to know is if there is a mechanism in photoshop that can monitor the export usage, we have not found it if it is there; is there a device that we can install on or in the printer that can number the images used per person...or be creative like the venerated Dr. Mark Farmer from the University of Georgia who suggested the "Jersey Method" which he claims to have developed in the mid 1980s.
Any help would be appreciated.
Funny colors at sunset, strange smells, dreary, damp, dank
John Grazul Rutgers University Electron Imaging Facility
The schedule of charges for the EM Unit must be documented and reviewed in accordance with the requirement of the (insert name eg EM Unit) Laboratory Manual.
The schedule of charges must be review by Management at {insert times}
Secondly, the path is to full cost recovery going hand in hand with ISO 9002 + GLP (in Aust AS Stnd) + (in Aust, NATA requirements).
My view again is of a Laboratory Manual + Laboratory Procedures + Operating Instructions (three levels).
Whenever a potential user contacts me, I ask if they have a reference with a method. We then try to follow that method if it is feasible and safe, simply so that our users can produce similar to the literature because this can save a lot of time in interpretation. If they only have old references then I may suggest that we prepare the sample with an up to date protocol and the original method. I keep a reasonable selection of specimen preparation texts/manuals and have the usual basic standard protocols.
All samples are then logged into our specimen book when they arrive in the lab with a unique specimen number which incorporates the year (eg 123/98), with the person's name, date of arrival in the lab, basic method (eg 2.5%G; 1%OsO4; in 0.1M cacod pH 7.2; ethanol; Spurr's) with details of the specimen including the outside lab's specimen numbers. If the details are brief you can also fit in a word or two of comment about the sample. This all fits on one line of two facing pages of an A4 book. If there is any substantial modification to a method this goes into the back of the same book with method and reference then the page of the method is written on the same line as the specimen details. It's compact, low tech, simple and cheap and the only flaw is that we must NEVER LOSE THE SPECIMEN BOOK (ie it doesn't leave the lab and we try to photocopy it from time to time.
This works well for the 300 or so samples that may come to us a year and is particularly useful for negative stains which we often need to adjust or modify slightly. The only samples that do not fit well into this system are those that have been prepared outside of the lab but I simply ask if the user has the details so that we can record them if they want and I make a note - processed elsewhere.
I still haven't decided what to do about the handwritten Millenium Bug ( ie300/99 -} 1/00 and beyond) but as electron microscopes, the lab and I may not be around when we come full circle, I don't think it's a problem. I may just get a smaller pencil to label my specimens.
Malcolm Haswell Electron Microscope Unit University of Suderland UK ----------
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Barry Searle wrote: =============================================== In search of........
Would someone have a copy of the SPI SIRA documentation that comes with the standard that I could have? ================================================ Sorry, we did not know you needed a set. We will FAX it to you immediately. Tell us what FAX number to FAX it to. We anticipate this information will also be up on our website shortly.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We have a fee schedule for package deals as well as individual parts of the project. We too allow user participation where tehy are competent and adjust prices accordingly.
I am attaching our fee schedule for "in-house" users. We recover only about 20-25% of the true costs. We are required to charge our rare external users for the full cost.
If you cannot read the attachment, let me know and I can fax to you
******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Assistant Director, The Biotechnology Program PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
I would suggest that you contact Tom Kubic at TAKA. I don't have a phone no. handy but know that they are on Long Island, in the Nassau County area (516 area code).
Best regards,
Barbara Foster
At 09:23 AM 5/6/98 -0500, Garber, Charles A. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are studying the origin of the two-length-scale diffraction phenomena in SrTiO3 single crystal (Phys.Rev.Lett.80,2370(1998)), and would like to chemical-polish or electro-polish the samples.
Does anyone have a recipe ? Any suggestion and comment are appreciated.
Thanks.
******************************** Dr. Yimei Zhu Materials Science Division Brookhaven National Laboratory Upton, Long Island, NY 11973 USA Tel. (516)344-3057 Fax. (516)344-4071 ********************************
for reference purpose I am searching for near-edge spectra (ELNES or XANES) of the K-edges of beta-Si3N4 (P6 3/m). If anybody know any publications concerning this structure, please email me directly.
Here is some information I have on the VCR Group. They also have an 800 number that I can't find right now.
Dale
VCRVINCE-at-aol.com
Vince Carlino VCR GROUP Incorporated 250 East Grand Avenue Ste. 70 South San Francisco, CA 94080
On Thu, 7 May 1998, Susanne Pignolet Brandom wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } If anyone has information on how to contact the VCR Inc in the USA, please } forward it directly to: } } Kai Tang } kaitang-at-us.imb.com } } Thank you } } Susanne } Susanne Pignolet Brandom Ph.D. } 239 Old Littleton Road } Harvard, MA 01451 } 978-456-3100 } email: spb-at-mwrn.com } } MicroWorld Resources and News http://www.mwrn.com/ } } }
Dale Shumaker G9 WBSB 725 N Wolfe Street Baltimore, MD 21205
Wilson Lab; Johns Hopkins University School of Medicine, Cell Biology and Anatomy 410-614-2654 410-955-4129 (fax)
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I'm working with material samples and I'm looking for a good TEM sample prep. class. I've been to the great class at GATAN and found it very helpful. Lehigh University is not offering a sample prep class this year. I would appreciate any other suggestions.. Thanks. D.McLean
My school has been offered a JEOL JEM 100U TEM. We may look for a non- profit buyer for this instrument. We would be grateful for information regarding its approximate value.
Hi, I wish to do some TEM on the embryo's of c.elegans, particularly at the 4 cell stage. I understand the shell is quite resilient and it has been suggested it would need to be cracked by a laser in glutaraldehyde. Does any one know of a protocol??
a copy of the Manual for the Concept EDM Model 111;
or
direct me to the current company (details of) handling this type of equipment - it was originally called Concept EDM Ltd, in England. I have no further details. This equipment is of the 1970's vintage.
We routinely do EM of c. elegans in our laboratory.
We mainly look at x-section. So what we do is cut both ends of the worm with razor blade, whick helps in infiltration. With embreyo's this is tough to do given its relative size. However, if enlongate all your normal processing protocol, and I have done that, you can get a with some good result.
Now, if you have laser, yes that's the way to go.
If you're interested in a processing protocol, I can supply you with the one I use.
On Tue, 5 May 1998, Patrick Merritt wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } I wish to do some TEM on the embryo's of c.elegans, particularly at } the 4 cell stage. I understand the shell is quite resilient and it has } been suggested it would need to be cracked by a laser in } glutaraldehyde. Does any one know of a protocol?? } } Thank You } } Patrick } } }
******************************************************** * Raj Patel * * Dept. of Pathology * * Robert Wood Johnson Medical School * * 675 Hoes Lane, Piscataway, NJ 08854 * * * * voice (732) 235-4648; Fax -4825 * * E-Mail rpatel-at-umdnj.edu * ********************************************************
Researcher in the area of "Electron microscopy in plant research". Reference number 1716/98-4599.
At the Electron Microscopy Unit, Department of Plant Breeding Research, The Swedish University of Agricultural Sciences, Sval=F6v/Alnarp, Sweden. The EM Unit will be a common resource for the university's departments in Southern Sweden.
The candidate must have a doctor's degree, primarily not earlier than five years ago. Experience of work with plant material is a merit. Of importance are the scientific, pedagogic, adminstrative and other capacities of relevance for this occupation. Of importance also is the capacity to inform about scientific research in a popular way.=20
The appointment is first for two years, followed by another period of two years and the possibility for a further prolongation. The salary is individually determined (at least 20 000 Swedish crowns a month, before taxes). Women are engouraged to apply.
Together with the application, a short account on the applicant's scientific and pedagogic activities should be given. The application marked with the above-mentioned reference number, together with a certified C.V., merit and publication lists, and other documents (including scientific and pedagogic works) should be sent in two copies so that they reach the "Registrator, SLU, Box 7070, S-750 07 Uppsala, Sweden" at the latest on June 5, 1998.
For further information contact professor Waheeb Heneen, tel +46 418 667064 or fax +46 418 667081.
Patrick Merritt wrote: } } Hi, } I wish to do some TEM on the embryo's of c.elegans, particularly at } the 4 cell stage. I understand the shell is quite resilient and it has } been suggested it would need to be cracked by a laser in } glutaraldehyde. Does any one know of a protocol?? } } Thank You } } Patrick
Patrick:
You might want to try "phase partition fixation" which has been successful on things like Drosophila eggs with a hydrophobic barrier. See Stain Technol. 52:89, 1977 or J. Histochem. Cytochem. 34:795-800; 1986.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
MME offers customized, on-site training in all areas of microscopy, including sample preparation. If you are interested in this alternative, please contact me directly.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
At 02:42 PM 5/7/98 -0600, Dorrance McLean wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Contact Don Riddle in the Dept of Biological Sciences at U. of Missouri-Columbia. His lab has been doing TEM of C. elegans for many years including serial sectioning. Phone # are 573-882-6363 or 882-2816.
Debby Sherman, manager Microscopy Center in Agriculture Purdue University W. Lafayette, IN 47907
--------------------------------------
Hi, I wish to do some TEM on the embryo's of c.elegans, particularly at the 4 cell stage. I understand the shell is quite resilient and it has been suggested it would need to be cracked by a laser in glutaraldehyde. Does any one know of a protocol??
Thank You
Patrick
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The phone # of VCR Group, Inc. is: 1-800-536-1827
Ya Chen
IMR U-Wisconsin
Ya Chen
======================================================================== \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL: 608-263-8481 \/ / / University of Wisconsin-Madison FAX: 608-265-4076 / / / 1675 Observatory Drive #159 / /__/_ Madison, WI 53706 Email: ychen14-at-facstaff.wisc.edu ======================================================================== IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
A user in our lab has started a project that requires the staining of many TEM grids, dozens, or more, at a time. She is really in mass production mode and is frustrated by keeping track of drops in dishes.
She saw a commercial automatic grid stainer ($10K, choke) and thought we should get it. That's kind of pricey for me to consider without some other feedback.
I would like to help her out. I am not sure most users in our lab (a fairly low volume central campus, general EM lab) would ever need an automatic stainer. Maybe you could give me some ideas about how useful and practical they are for routine use.
If they are not what we need, is there anything I could get to help her with this project? How useful are the little gizmos in some of the catalogs? What is your favorite? Have you tried any of them and been happy, or not?
Thanks
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Maybe one of you can save me turning my office upside down looking for info. on embedding and sectioning rocks for TEM. I know I have all this stuff somewhere but its location is somewhere deep in my memory banks.
I have a user who wishes to look at pieces of rock from a diamond anvil cell experiment.
The piece is very small, but not thin enough to go directly into the TEM. We are trying lots of things to prepare the sample, sectioning is next. I know how to section and the small size of the sample is no problem. I just can't remember if there are any tricks or recommended procedures for getting a little piece of rock to infiltrate and stick in the plastic. While I'm asking, any plastic or a special one?
Any other ideas on how to help on this project would be wonderful. As always your kindness and sage advice are much appreciated
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
I am a new SEM user and I would be very grateful for any suggestions to the following problem. I have microbeads that cells have been cultured on, they adhere tightly to the outer surface of the bead. I examine them at lower powers (X1,000 or less) with the SEM, all is well until I dry them down (I use coverslips) the beads/cells are so light and fragile that after gold coating they just do not 'stick' to the coverslip and the slightest movement sends them all over the coverslip. I would like to find a way to make them adhere to the slide but still look 'clean'. Thank you for any help, Gill
Gillian Rittman Research Associate, University of Texas - Houston Research Office 4.109 DBB 6516, John Freeman Ave, Houston, TX 77030. Phone (713)500-4359 FAX (713)500-4372
Hi everyone, I am looking for any information on chemically etching nylon. I have a nylon barrier which has low adhesion on one side, and really good adhesion on the other. The manufacturer swears that the two side of the barrier are the same and the adhesion systems are the same. I would prefer not to microtome the sample so I was wondering about etching followed by examination by SEM.
{fontfamily} {param} Geneva {/param} {bigger} {bigger} I am in the process of updating the MSA SEM Outreach list for the Education Committee. This particular list will include anyone who has some kind of education outreach program which involves electron microscopes, especially scanning electron microscopes.
The information I need is: Name, address, phone number, fax number, e mail address, what kind of program do you have (lab tours, hands on microscope operation, in school seminars and lab work, etc.), and what age students are involved (high school, jr. High school, elementary students, etc.)
Please send your response to my e-mail address: gardnerj-at-acs2.byu.edu
I look forward to hearing from you. Thanks for your assistance.
} I have a user who wishes to look at pieces of rock from a diamond anvil } cell experiment. We are trying lots of things to prepare the sample, } sectioning } is next. I just can't remember if there are any tricks or } recommended } procedures for getting a little piece of rock to infiltrate } and stick in the } plastic. While I'm asking, any plastic or a special one?
} Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA Hi, Jon -
"Rock" isn't very helpful. Porosity makes a big difference. So does mineral content & Moh hardness. If it's a bit porous, try LR White, hard grade. I once gave that advice to a postdoc with a copper-containing mineral, and the stuff dissolved in the LR White overnite, making it a beautiful blue. Araldite worked well enough for her that she got the Diatome award from MSA (and her mom was living in Switzerland!). Look at these refs:
Csencsits, R., Schooley, C., and Gronsky, R. (1985) An improved method for thin sectioning of particulate catalysts. J.E.M. Technique 2:643-644. Ulan, J.G., Schooley, C., & Gronsky, R. (1990) Microtomy of large particle zeolites for TEM. Mat. Res. Soc. Symp. Proc. 199:153-156 Ulan, J.G., Schooley, C., & Gronsky, R. (1990) Modified embedment procedure for microtomy of large particle zeolites. J.E.M. Technique 16:254-255
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Frank Karl wrote: ======================================================= I am looking for any information on chemically etching nylon. I have a nylon barrier which has low adhesion on one side, and really good adhesion on the other. The manufacturer swears that the two side of the barrier are the same and the adhesion systems are the same. I would prefer not to microtome the sample so I was wondering about etching followed by examination by SEM. ======================================================== You did not mention which nylon specifically you are working with, but we have found one "quick and dirty" way to look at the question of asymmetries of films is to look at the two surfaces by Pt/C surface replication and TEM, and then see to what degree the two sides really are different. Indeed very rarely have we found the two different surfaces to be the same. So your thought that the two sides might in fact not be the same could very well be quite correct.
But if you want to etch one surface in a controlled way, why not try plasma etching? I am talking about isotropic etching (as opposed to anisotropic etching) using oxygen in a small barrel table top unit, like our own SPI Plasma Prep II unit or ones made by several other firms such as Denton Vacuum. A polycarbonate membrane filter, with oxygen, is etched completely away in about 30 minutes and I would expect any of the nylons to etch at approximately the same rate. It sounds to me you are talking about a one or two minute etch to remove that surface barrier layer you think is present.
More information about plasma etching can be found on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
RMC, Tucson, AZ, USA will hold it's 5th Materials Science Ultramicrotomy Course on September 29-October 2, 1998 in Tucson.
This course covers all aspects of microtomy of materials for TEM, SEM, SP= M, and LM including: types of resins, how to choose; all types of knives and their selection; etching, staining, sectioning of polymers of all Tg's; sectioning or preparing polished SPM/SEM faces on hard materials such as semiconductors= , metals, ceramics.
The emphasis of the course is morning lectures followed by afternoon lab sessions where students actually DO the work, not just watch it demonstrated. You will return to your lab and be able to do the work you= r organization needs to attack difficult geometry samples like particulates=
and fibers. You will know how to select from all the different resins and=
knives. You will see how to use microwave processing for 2 hour turnarou= nd on sample embedding when speed is important.
The course price includes manual, lab supplies, tuition, hotel, meals and=
banquet night for $1950 USD.
Please see our web page at WWW.RMC-Scientific.com/microtomes/
For a course announcement please email to: RMC-at-RMC-Scientific.com attn: A= nn Nadeau or call 520-903-9366, Fax 520-903-0132
Responding to the message of {199805081759.KAA29444-at-cats.ucsc.edu} from jmkrupp-at-cats.ucsc.edu (Jon Krupp): } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } A user in our lab has started a project that requires the staining of many } TEM grids, dozens, or more, at a time. She is really in mass production } mode and is frustrated by keeping track of drops in dishes. } } She saw a commercial automatic grid stainer ($10K, choke) and thought we } should get it. That's kind of pricey for me to consider without some other } feedback.
SNIP
SNIP
} How useful are the little gizmos in some of the } catalogs? What is your favorite? Have you tried any of them and been happy, } or not?
Some of the gizmos are very useful and inwexpensive. The one I like to use is the Synap Tek GridStick kit, available from Ted Pella, for around $25. Kit includes 5 gridstick bars which hold 11 grids each in a simple pipet. Only small volumes of stains are required, almost no contact with air during staining, can do rinses with them, easy to use and they don't make a mess. I don't use the flow-limiting plugs that come with them, tho. Just be careful to not intake or expell liquids too fast, to avoid turbulent flow. These work great for staining ultra-thin epoxy or acrylic sections, not sure sure about thick sections.
Another kit that allows you to process even more grids simultaneously, is the Hiraoka Grid Staining Kit, available from Polysciences. Its does up to 40 grids at once, and uses a trough for stains over which you invert a plastic square with slots in that hold the edge of the grids by pressure. I've not used it as I never have to do that many grids at a time, care is required to avoid a mess, but any careful, patient, motivated person who needs to stain lots of grids would be able to use it successfully.
I have no financial interest in Pella or Polysciences, just a satisfied user of these two products.
I'd like to hear about what others use as alternatives to the ol' classical method of drops on Parafilm surrounded by a ring of sodium hydroxide pellets, inside of a covered Petri dish.
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
Jonathan Krupp wrote, with regard to grid stainers: =============================================== ..............How useful are the little gizmos in some of the catalogs? What is your favorite? Have you tried any of them and been happy, or not? =============================================== An often overlooked publication, Microscopy Research and Technique 26:177- 179 (1993), by Ming H. Chen, Medicine/Dentistry Electron Microscopy Unit, Surgical-Medical Research Institute, 1074 Dentistry Pharmacy Bldg., U. of Alberta, at least from my perspective, describes the ultimate "gizmo". It holds up to 100 grids and was designed specifically to reduce the amount of expensive immuno reagents needed to "prime" it and get it working. It can be operated with a volume as small as 2 ml in fact.
We were so impressed with this little "gizmo" that we have been offering it as our "SPI Stain 'n Wash™ Grid Staining System" for some several years. It is in use in a number of laboratories worldwide. It is fully described with a number of photos and other graphics on our website below. And it is very low cost.
Hope this is not too commercial sounding. But if one is working especially with expensive reagents, this is a really useful (and money saving) item.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Jon, We have an LKB ultrostainer in our lab, I think now supplied by Leica. We cannot recommend it enough!, especially in a multiuser facility like ours, (and yours). Can stain 38-40 grids at a time, low contamination of sections, no exposure to nasty chemicals and very easy to use. The small plates which come with the stainers only hold about 20 grids, we actually buy one from another supplier which holds 40, which is better for us (Hiraoka staining plate). The only problem sometimes, especially with our end of the world is supply of the stain bags. But over i your neck of the woods, it probably isnt too bad. We calcualted I think, that in the first 2 years of use, it had saved about 6months worth of staining time!
Hope this is of some help, as I mentioned, if you can get one, and you have lots of grids, go for it! OUrs is about 10-12 years old now, and have had minimal troubles with it. Not like some of the newer equipment we have recently purchased in our lab!
All the best,
Rich.
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
We use the flexible plastic piece from the Hiraoka kit, but put a large puddle of stain over the sections rather than inverting the holder over the square dish of stain. For washing we support the plastic by a wire frame over a large beaker, and dribble water over it through a Pasteur pipette connected through a hose to a water container.
Sally Stowe
---------------------------------------------------------------------- Dr Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: Box 475 ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 (0)2 6249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, Australia 2601 http://online.anu.edu.au/EMU/home.htm |AUSTRALIA 0200
We have been given the go ahead to upgrade our electron microscope preparation laboratory area including the choice of installing new fumehoods and/or biohazard cabinets.
At present we have two fume hoods and no biohazard cabinets. We are thinking of asking for four fume hoods or, alternatively, three fume hoods and one biohazard cabinet. Our feelings were that the biohazard cabinet might be safer considering the human biopsy material we deal with, but maybe it would be no safer than a good fume hood.
My question is: should we consider a biohazard cabinet in place of a fumehood given that many of the biohazards dealt with in tha lab are also in toxic fixatives or solvents? Do other EM labs use biohazard cabinets in preference to fume hoods?
(By fume hoods I mean that the fumes are extracted from the room and released outside whereas a biohazard cabinet filters the air and returns it to the room).
Many thanks for any thoughts you might have on this matter.
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
With reference to the problem of etching nylon. Get hold of thr book "Polymer Microscopy" by Linda Sawywe and David Grubb Chapman and Hall 1987 ISBN 0-412-25710-6. It's got everything you would ever want to know about polymers microscopy.
Patrick Echlin Multi-Imaging Centre School of Biological Sciences University of Cambridge
On Fri, 8 May 1998 fskarl-at-goodyear.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi everyone, } I am looking for any information on chemically etching nylon. I have a nylon } barrier which has low adhesion on one side, } and really good adhesion on the other. The manufacturer swears that the two } side of the barrier are the same and the adhesion systems are the same. I } would prefer not to microtome the sample so I was wondering about etching } followed by examination by SEM. } } Thanks } } Frank Karl }
Dear Colleagues, will you please let me know if you happen to have a chemical formula for Vestopal-W?
I would like to calculate the average value of Z^2/A needed in quantitative electron microprobe analysis (Z=atomic number, A=relative atomic weight). I have embedded some organic compounds in Vestopal-W (to be used as concentration standards).
I am doing ultrastructural localisation of a mitochondrial membrane protein using ultramicrotomy and immunogold labelling with human liver biopsy specimens;fixed with formaldehyde(4%) and glutaraldehyde(0.1%) in phosphate buffer pH7,4.My basic problem is that I do not get good preservation of the mitochondria and the tissue appears to be scattered; though the labelling seems to be working.Please help me if you have done some work with liver tissue. Kwanele B. Siziba Email:kwanele-at-uctgsh1.uct.ac.za
Dear colleagues, will you please tell me what is the chemical formula of Vestopal-W? I would like to calculate Z^2/A factor needed for electron microprobe X-ray analysis of Vestopal-embedded material. Thanks very much, Sincerely, Radek Pelc (Mr.) (Prague)
P.S. I use my lab colleague's account to reach Microscopy discussion. +---------------------------------------------------------------+ Oldrich Benada Acad. Sci. CR Phone: +420-2-4752399 Institute of Microbiology Fax: +420-2-4715743 Electron Microscopy Group E-mail: benada-at-biomed.cas.cz Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +---------------------------------------------------------------+
I would go with one biocabinet and rest fume hoods. You mention the tissue you deal with: once they are fixed in glutaraldehyde (formalin) they no longer need to be handled as biohazardous ( in the real world). But they do need to be handled in a fume hood after fixation. The only time I can think when a sample needs to be completely handled in a biohood, is with unfixed viral or bacterial negative stain procedures. In a past job, our lab was remodeled which included a biohood, I only used it to store things in, never as a biohood.
Best of Luck, Ed Calomeni Dept. Pathology Medical College of Ohio 3000 Arlington Ave. Toledo, OH 43614
419-383-3484 ecalomeni-at-mco.edu
} } } Richard Easingwood {richard.easingwood-at-stonebow.otago.ac.nz} 05/10 10:41 pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello microscopists,
We have been given the go ahead to upgrade our electron microscope preparation laboratory area including the choice of installing new fumehoods and/or biohazard cabinets.
At present we have two fume hoods and no biohazard cabinets. We are thinking of asking for four fume hoods or, alternatively, three fume hoods and one biohazard cabinet. Our feelings were that the biohazard cabinet might be safer considering the human biopsy material we deal with, but maybe it would be no safer than a good fume hood.
My question is: should we consider a biohazard cabinet in place of a fumehood given that many of the biohazards dealt with in tha lab are also in toxic fixatives or solvents? Do other EM labs use biohazard cabinets in preference to fume hoods?
(By fume hoods I mean that the fumes are extracted from the room and released outside whereas a biohazard cabinet filters the air and returns it to the room).
Many thanks for any thoughts you might have on this matter.
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
I take rings-shaped slices of BEEM capsules(slices are about 4-5 mm high); cut perpendicular slits, evenly spaced, along one side of the ring, leaving room on the other side for pinching the ring ,which opens the slits. Then you place grids into the slits, grabbing just the rim of the grid. When you let go of the pinching fingers, the grids are held in place. It takes a bit of practice to get the slits spaced right and to learn how to get several grids into the slits without losing the first ones you put in. Then I stain using 10ml beakers, submersing the grid rings in the stain, and rinsing by bobbing the rings up and down in water, holding the rings with tweezers. You can blot excess water between the wet grids with points of filter paper.It takes more stain but I think the results are cleaner. Sometimes I produce a "lucky" ring that will hold 6 grids for me!
Julie Gross Dept. of Anatomy UCONN Health Center Farmington, CT 06029
I would appreciate hearing from anyone who has experience using=20 Tetrahydrofuran (THF) for dissolution of PVC filter membranes and=20 redeposition of particules remaining in the THF solution onto silver=20 membrane filters=2E This is done by vacuum filtration=2E =20 =20 What types of vacuum pumps can safely be used for this purpose and can= =20 THF be sonicated=2E =20 =20 Need to know safety issues such as handling, processing issues as=20 above, storeage and disposal of waste=2E We are assuming that all wor= k=20 will be done in a fume hood=2E =20 Thanks for any assistance =20 John Humenansky Braun Instertec 6875 Washington Ave=2E So=2E Minneapolis, MN 55439 612-942-4822
The number you have for Concept EDM is correct, it is also their fax number +44 (0) 1628 639854. I don't use them but a quick call was answered this afternoon (UK time).
Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Gill, I have processed cells on beads a while ago. I used to not attached them to coverslip until I dried them ( I would process them in folded , stapled filter paper). After drying I would pick them up on stubs with sticky tape on it and then metal coat them. Hope this helps, Lilith
I am a new SEM user and I would be very grateful for any suggestions to the following problem. I have microbeads that cells have been cultured on, they adhere tightly to the outer surface of the bead. I examine them at lower powers (X1,000 or less) with the SEM, all is well until I dry them down (I use coverslips) the beads/cells are so light and fragile that after gold coating they just do not 'stick' to the coverslip and the slightest movement sends them all over the coverslip. I would like to find a way to make them adhere to the slide but still look 'clean'. Thank you for any help, Gill
Gillian Rittman Research Associate, University of Texas - Houston Research Office 4.109 DBB 6516, John Freeman Ave, Houston, TX 77030. Phone (713)500-4359 FAX (713)500-4372
A while ago I posted a question as to which is the best program for doing morphometry on bone sections. There were few programs suggested (Bioquant being the most popular one). After studying the programs we found out that neither of them count trabeculae. We are doing it by hand and it is a lot of work. We haven't purchased a system yet and are still hoping to find a program that will cover all our needs (which are the most common ones) including the trabecular count. Thanking you in advance, Lilith
Responding to the question of Richard Easingwood {richard.easingwood-at-stonebow.otago.ac.nz}
{My question is: should we consider a biohazard cabinet in place of a {fumehood given that many of the biohazards dealt with in tha lab are {also {in toxic fixatives or solvents? Do other EM labs use biohazard cabinets {in {preference to fume hoods?
{(By fume hoods I mean that the fumes are extracted from the room and {released outside whereas a biohazard cabinet filters the air and returns {it {to the room).
There are several types of biosafety cabinet you should consider for use with toxic vapours. The basic Class I (negative pressure) pulls air in and upwards inside the hood and works in the same way as a chemical fume hood. The exhaust goes through a hepa filter and is exhausted outside the building. This has the same problems as conventional fume hoods, when using heavy vapours that are denser than air, you need enough draught to keep them in and flowing upwards. The disadvantage of this is that the inside is not a sterile environment for tissue culture etc.
The class II biosafety hoods have a vertical laminar flow. Thus contaminated air is drawn downwards, through a mesh at the front, under the worksurface and upwardsthrough ducting at the back of the hood. Some of these type IIs vent hepa filtered air back into the lab, others vent to the outside and are therefore suitable for use with toxics and volatiles. The other main difference is whether you want to keep the inside sterile for cell culture or if you just want to keep biohazards and solvents away from the operator. Because of the downward draft in front of the worker, a type II B2 biosafety cabinet was chosen for our new EM lab. Heavy vaour will tend to go straight down into the duct. This is also suitable for tissue culture since the air inside remains sterile. If you are just going to fix things in the hood, then you only need a class I hood or a class II B1 hood neither of which filter the air entering the workspace, and are therefore less expensive, and might also give higher air flow rates. However, if you want to take samples from a tissue culture at various time intervals, you want to keep the inside strerile as well.
As with all hoods, the actions of operator can interfere with the air flow causing the protection to be less than 100 per cent. If you need to use large amounts of solvents, obviously a dedicated fume hood tends to have better air flow rates as there is no requirement for a laminar flow and also the airflow is not slowed down by passing through a HEPA filter. For this reason we also got two conventional chemical fume hoods as well.
Dr Timothy F. Booth Canadian Food Inspection Agency National Centre For Foreign Animal Disease Suite T2300 1015 Arlington St. Winnipeg Manitoba R3E 3M4 CANADA http://www.hc-sc.gc.ca/main/lcdc/web/bmb/fedlab_e.html#toc email tbooth-at-em.agr.ca Tel 204 789 2022 Fax 204 789 2038
We also use the UltroStainer and are very happy with it. It is now about = 5 years old and receives daily use. Although we did have a problem with = it while it was relatively new, it has not given us any problems since. = It is reliable, clean, and almost maintenance free. Several years ago = there was problems with the stain bags, but the company seems to have = fixed that now. One caution about autostainers, they do stain both sides = of the grid. And I am not convinced that this is the cheapest way to = proceed.
Normal disclaimers apply: no financial interest, and the opinions = expressed are my own.
} } } Jon Krupp {jmkrupp-at-cats.ucsc.edu} 2:05:01 PM 5/8/98 } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html=20=
A user in our lab has started a project that requires the staining of many TEM grids, dozens, or more, at a time. She is really in mass production mode and is frustrated by keeping track of drops in dishes.
She saw a commercial automatic grid stainer ($10K, choke) and thought we should get it. That's kind of pricey for me to consider without some other feedback.
I would like to help her out. I am not sure most users in our lab (a = fairly low volume central campus, general EM lab) would ever need an automatic stainer. Maybe you could give me some ideas about how useful and practical they are for routine use.
If they are not what we need, is there anything I could get to help her with this project? How useful are the little gizmos in some of the catalogs? What is your favorite? Have you tried any of them and been = happy, or not?
Thanks
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu=20
******************************************************* I am looking for any information on chemically etching nylon. I have a nylon barrier which has low adhesion on one side, and really good adhesion on the other. The manufacturer swears that the two side of the barrier are the same and the adhesion systems are the same. I would prefer not to microtome the sample so I was wondering about etching followed by examination by SEM. ********************************************************
Nylon is a "swine" as regards liquid chemical etching of any kind. The problem is that nylons are soluble in acid. Some people claimed to have "etched" nylon with formic acid, but what they have in fact done is to swell the surface layer into jelly, which has recrystallized once the formic acid is removed.
There are gentle solvent treatments which do not actually dissolve the polymer, but differentially swell the crystalline and amorphous layers. See, for example,
Bartosiewicz,I & Mencik,Z. J.Polym.Sci. Polym.Phys.Edn. 1974, v 12. pp 1163-75
But more severe solvent treatments can produce recrystallized layers with spurious morphology.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I also concur with Sally's Hiraoka kit, which we also use in the same manner. Except that to wash the grids, we first dump the stain, and then grab the plastic with locking hemostats and then vigorously shake it up and down in water through 3 beakers of water, 60 times per beaker. In order to ensure that the sections don't float off, we first dry the sections on the grids for 5 minutes in a drying oven. Then there is no force in the universe which can remove the sections.
For the Lead Citrate stain, we use a glass petri dish with 4 KOH pellets to remove the CO2. We hold the Hiraoka a bit higher by supporting it on a plastic frame used for paraffin embedding.
The method works well.
Garry
} We use the flexible plastic piece from the Hiraoka kit, but put a } large puddle of stain over the sections rather than inverting the } holder over the square dish of stain. } For washing we support the plastic by a wire frame over a } large beaker, and dribble water over it through a Pasteur pipette } connected through a hose to a water container. } } } Sally Stowe } } } ---------------------------------------------------------------------- } Dr Sally Stowe |Email: stowe-at-rsbs.anu.edu.au } Facility Coordinator |Post: Box 475 } ANU Electron Microscopy Unit |ANUEMU (RSBS) } Ph 61 (0)2 6249 2743 |Australian National Univ. } FAX 61 6 249 4891 |Canberra, Australia 2601 } http://online.anu.edu.au/EMU/home.htm } |AUSTRALIA 0200 } } }
Polysciences used to make a device called the "Multiple Grid Staining Unit" it would stain 24 grids at a time it took about 8 mls of stain and was very handy. They quit production but keep promising to make it again. It's on page 85 of their 95-96 catalog. I figure if enough people call them & ask them about it they'll realize there is a demand and make them again. It's a very neat device and makes life easy for the lab that stains a lot of grids at one time. So give 'em a call & let's get this thing made again!
Paula :-)
p.s. I have no other interest in this than wanting to buy new ones to replace the ones we have that are wearing out.
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Does anyone out there have a glass knifemaker that they no longer use and are willing to part with for a modest cost...or for free if I pay shipping? I am looking for either an LKB model 7801A or 7801B, or other manufacture that will take the standard 6.4x25x400 millimeter dimensions glass bar stock.
This knifemaker, along with one of our ultra-microtomes, will be donated to a project in Morocco that works on plant virus diseases using TEM as a tool.
Thanks for any assistance you can give,
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
I use the SynapTek grid staining system, also, to stain dozens of grids. To overcome the problem of turbulence created in the staining pipettes, a clever person who preceeded me melted down the pipette tip, presumably with a Bunsen burner, to create a larger opening at the tip's end.
Missy Josephson Eleanor Josephson Department of Anatomy MC-3405 263 Farmington Ave. Farmington, CT 06030-3405
What are the imaging issues in your trabeculae application? Is it the three-dimensionality, their size, the ability to segment just the spaces? There are several very good programs on the market. If you can email me a file or two, maybe I can make a suggestion.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
At 09:36 AM 5/11/98 -0400, Barry, Lilith wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Looking for standards for TEM image and diffraction calibration that are tracable to NIST for certification purposes. We would prefer to just purchase ready made specimens.
Thanks,
Mark A. Wall L-350 Lawrence Livermore National Lab C&MS dept. 7000 East ave Livermore,CA USA 94550
The standard you need is the MAG*I*CAL TEM Calibration Standard.
The MAG*I*CAL is a TEM calibration standard that performs all of the thr= ee major instrument calibrations for a TEM: image magnification; camera constant for indexing diffraction patterns; and image/diffraction patter= n rotation for relating crystal directions to features in the image. =
MAG*I*CAL consists of an electron transparent cross-sectional TEM sample made from a MBE grown, single-crystal semiconductor wafer. When the calibration structure is viewed in a TEM, it appears as a series of light=
and dark layers where the layer thicknesses are accurately known. The calibrated thickness measurements of these light (silicon) and dark (SiGe=
alloy) layers are based on careful TEM measurements of the {111} lattice=
spacing of silicon which is visible on the calibration sample itself, and=
are supported by x-ray diffraction measurements. The layer spacings are designed so that the sample can be used to calibrate the entire magnification range in a TEM - from 1,000X to 1,000,000X. As the sample = is also a single crystal of silicon, the calibrations requiring electron diffraction information such as the camera constant and image/diffraction=
pattern rotation can also be performed easily and unambiguously. One single calibration sample can therefore be used to provide all three of t= he major TEM instrument calibrations at all magnifications and all camera lengths.
With regard to the traceability and certification of the MAG*I*CAL(TM) calibration sample, each sample is grown on {001} oriented single crystal=
silicon, and all spacings on the sample are directly referenced to the =
cross-sectional (111) lattice spacing of silicon. This spacing is visibl= e by lattice imaging on the sample itself, giving each sample the capabilit= y of being self-calibrating. Each unit comes with a numbered certificate,=
the =
text of which is included below. This certificate has been used for ISO 9000 certification, with the argument that to our knowledge, this is the highest quality TEM sample available anywhere in the world at this time. = =
The MAG*I*CAL (TM) calibration sample consists of sets of thin, nominally=
10 nm alloy layers of Si0.81Ge0.19 alternating with 10 nm pure silicon layers, on a single crystal silicon {001} substrate. These electronic device quality layers were grown by Molecular Beam Epitaxy (MBE) as strained layers, i.e., the alloy layers have a slightly different crystal=
lattice constant, but are strained to conform to the lattice spacing of pure silicon, so that the material remains single crystal. Lattice image= s should therefore be taken in the region of the sample containing no Ge, b= ut other measurements are unaffected. The layer thickness variation across t= he wafer was measured by double crystal x-ray diffraction (DCXRD) mapping as= { 1.0%. =
All four sets of the five thin Si0.81Ge0.19 alloy layers and alternating pure silicon layers (superlattices) were directly calibrated by high resolution transmission electron microscopy (HREM) with the cross-section= al (111) =
lattice spacing of the single crystal silicon substrate, equal to 0.31354= 3 nm [1]. These measurements are also supported by (DCXRD). =
The error in all spacings in the superlattices is one atomic layer:
=A6t =3D +0.3 nm or approximately +3% The larger, nominally 1.0 micron silicon spacings were calibrated against=
these superlattices. The total error across the entire calibration sampl= e is given as:
=A6t =3D + 3%
[1] CRC Handbook of Chemistry and Physics, CRC Press, Inc., Boca Raton,=
Florida 33431
South Bay Technology, Inc. supplies the MAG*I*CAL and so I have a defini= te financial interest in promoting its use. I also have copies of other research papers that have been written by the developer, John Mccaffrey, which will provide you with much greater detail. If you have an interest= , please let me know and I'll forward the information to you.
As a matter of additional interest, we can provide batches of the MAG*I*C= AL which are all made from the same wafer which provides the ultimate in calibration uniformity. This has proven to be an ideal solution to large=
organizations who are looking for a "company standard" calibration technique. Please inquire for more information on this service.
I think this should suit your requirements. If you require any additiona= l information, please feel free to contact me.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Mark Wall } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Looking for standards for TEM image and diffraction calibration that are tracable to NIST for certification purposes. We would prefer to just purchase ready made specimens.
Thanks,
Mark A. Wall L-350 Lawrence Livermore National Lab C&MS dept. 7000 East ave Livermore,CA USA 94550
Hi, I am looking for an inexpensive used SEM. Must have EDX or WDX capabilities. If anyone know of one for sale please e-mail me. Thanks Sincerely, Ijaz Rauf
*******************DO NOT REPLY DIRECTLY TO THIS E-MAIL******************** ****************USE THE ADDRESS LISTED AT THE END OF THE AD****************
UES, Inc. Air Force Research Laboratory Materials & Manufacturing Directorate Microstructural Characterization Facility WPAFB Dayton, Ohio
POSITIONS AVAILABLE
Microprobe/OIM Scientist Application of analytical scanning electron probe techniques to microstructural problems in materials science. Experience with electron microprobe analysis and scanning electron microscopy in conjunction with wavelength dispersive x-ray spectroscopy, energy dispersive x-ray spectroscopy, and orientation imaging microscopy. Minimum of a M.S. in Materials Science and 5 years of professional experience
TEM Scientist Application of basic analytical electron microscopy and conventional TEM techniques to microstructural problems in materials science. Experience with parallel electron energy loss spectroscopy, energy dispersive x-ray spectroscopy and convergent beam electron diffraction analysis as well as selected area diffraction analysis, bright-field, dark- field, weak-beam microscopy. Minimum of a B.S. in Materials Science and 2 years of professional experience.
Metallography/OM Scientist Application of metallographic and optical microscopy techniques to microstructural problems in materials science. Experience with metallographic preparation and subsequent microstructural analysis, via optical microscopy, of high temperature ceramic, intermetallic and metallic materials. Minimum of a B.S. in Materials Science and 2 years of professional experience.
To meet requirements for the Joint Comission, I have been asked by the laboratory manager to come up with Age Specific Competencies as part of our Position Description Format for each employee. To my knowledge, there are no age specifiic competencies for histotechs as they have no patient contact.
Can anyone help me with this? Thanks!!
Kathy Pelton -Henrion Supervisor of Histology SUNY HSC at Syracuse
} } I would appreciate hearing from anyone who has experience using } Tetrahydrofuran (THF) for dissolution of PVC filter membranes and } redeposition of particules remaining in the THF solution onto silver } membrane filters. This is done by vacuum filtration. } } What types of vacuum pumps can safely be used for this purpose and can } THF be sonicated. } } Need to know safety issues such as handling, processing issues as } above, storeage and disposal of waste. We are assuming that all work } will be done in a fume hood. } John -
There are a few OSHA procedures for preparing air samples collected on PVC filters for analysis by x-ray diffraction. If I recall correctly, both the crystalline free silica and the vanadium pentoxide methods suggest using THF as a solvent for dissolving the filter. My understanding is that not all of the PVC filters available on the market can be prepared using this method.
Mike Rose at OSHA's Salt Lake City Technical Center has done a great deal of work in this area. You may want to consider contacting him to discuss this in greater detail. I think he can be reached at 801-487-0267 or you may be able to place an inquiry at their web site {http://www.osha-slc.gov} .
Keith Rickabaugh Manager, Materials and Particle Characterization {krickabaugh-at-rjlg.com}
RJ Lee Group, Inc. 350 Hochberg Road Pittsburgh, PA 15146 ph: 724-325-1776 {www.rjlg.com}
Does anyone out there have, or know of, a used cryostat microtome for sale?
It does not have to be a late model; an old mechanical one in good working order (including the refrigeration unit) is all that is needed. IEC, TissueTek, Jung, etc., ok., need to use at minus 20 C.
Prefer someone in the southern hemisphere - anyone in New Zealand or Australia, but might consider shipping further.
Please respond to me directly.
Thanks in advance,
Heather Heather K. Smith, Ph. D. Dept. of Sport and Exercise Science University of Auckland Private Bag 92019 Auckland, New Zealand
Jon- there used to be two type of multi stainers on the market, I've used:
1) Hiroka stainer- basicly it is a soft piece of plastic with small slits cut in it, when you bend the plastic the holes open and release grids, when flat holes close and hold grids. flip it upside down in staining solution ... and usually all grids stay in and get stained. 2) Polysciences (I think) multi grid stainer, basicly a grid box with holes in the back side of the holder (inner part) and holes on the front side of the cover (outer part) these work very well, and can probably be made quite easily with the appropriate drill bits and some grid boxes...
good luck -Mike
On Fri, 8 May 1998, Jon Krupp wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } A user in our lab has started a project that requires the staining of many } TEM grids, dozens, or more, at a time. She is really in mass production } mode and is frustrated by keeping track of drops in dishes. } } She saw a commercial automatic grid stainer ($10K, choke) and thought we } should get it. That's kind of pricey for me to consider without some other } feedback. } } I would like to help her out. I am not sure most users in our lab (a fairly } low volume central campus, general EM lab) would ever need an automatic } stainer. Maybe you could give me some ideas about how useful and practical } they are for routine use. } } If they are not what we need, is there anything I could get to help her } with this project? How useful are the little gizmos in some of the } catalogs? What is your favorite? Have you tried any of them and been happy, } or not? } } Thanks } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } } }
We have gradually aquired a cloud around the outer edge of our negatives processed in an Arkay developer. The problem does not occur in trays or when processing a single hanger. When you process three hangers all nine negatives are cloudy.
We cleaned the developer tank with old fixer and ferricyanide, the hangers and fixer tank with 5% nitric acid and scrubbing. We made up new solution. And the problem continues.
We cannot seem to locate Omega-Arkay. Any advice or location of Arkay is appreciated.
The only calibration standard which I know of that does both image and calibration is the MAG*I*CAL from South Bay Technology. Try David Henriks: PH: 714-492-2600 Website: www.southbaytech.com e-mail: henriks-at-southbaytech.com They will be covered in the upcoming July article in Am. Lab: "Focus on Microscopy: What's New at Microscopy & Microanalysis '98".
Moxtek also makes electron microscopy standards. Try Doug Hansen at (801)220-0930 (Orem, UT).
I think both offer NIST calibration services.
Disclaimer: MME is not commercially involved with either of these companies.
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
At 11:26 AM 5/11/98 -0700, Mark Wall wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I also use the Hiraoka Grid stainer (use caution when removing it from the plate holder or grids can pop out). Instead of the staining container that comes with the kit I use disposable plastic weigh boats for holding both the stains and water rinses. You can put the entire unit inside of a glass petri dish with NaOH pellets to remove CO2 during lead staining. The weigh boats measure 4 cm on each side and hold about 8 mls. I often use a small weight to hold the flexible plastic grid holder down so that no air gets in. JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94022
The bone morphometry research in the fluorosis on the x ray radiograms was made at the Foundry Reseach Institute in Krakow Poland by me and dr Edward Czerwinski from Collegium Medicum - Ortopedic Departmenet Jagiellonian University - Krakow. I used for the x-ray bone analysis special program made by me on the Quantimet 570 with macroviver or microscopy.
The more medical data from this research was publish at the medical report.
About this problem write to;
Dr Edward Czerwinski Collegium Medicum - Head Ortopedic Department Jagiellonian University ul. Kopernika 19a, 31-501 Krakow Poland fax +48 12 4214046
best regards for all
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
On Mon, 11 May 1998, Barry, Lilith wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } A while ago I posted a question as to which is the best program for doing } morphometry on bone sections. There were few programs suggested (Bioquant } being the most popular one). After studying the programs we found out that } neither of them count trabeculae. We are doing it by hand and it is a lot of } work. We haven't purchased a system yet and are still hoping to find a } program that will cover all our needs (which are the most common ones) } including the trabecular count. } Thanking you in advance, } Lilith } } Lilith Ohannessian-Barry } NRC, IBS, } Ottawa, Ont. K1A 0R6 } Tel;613-993-6460 } Fax;613-941-4475 } e-mail; lilith.barry-at-nrc.ca }
On Mon, 11 May 1998 jhumenansky-at-brauncorp.com wrote:
} I would appreciate hearing from anyone who has experience using } Tetrahydrofuran (THF) for dissolution of PVC filter membranes and } redeposition of particules remaining in the THF solution onto silver } membrane filters. This is done by vacuum filtration. } } What types of vacuum pumps can safely be used for this purpose and can } THF be sonicated.
We have often filtered suspensions in organic solvents onto membrane filters in order to observe the particulates. We find a simple Buchner type water pump sufficient. The vacuum may not look good compared to a mechanical pump, but in terms of simple pulling power it is about 90% as strong.
We have used a Millipore or Sartorious demountable 25 mm membrane filter apparatus - the Neoprene bung at the bottom simply corks into the mouth of the Buchner flask. When filtration is done, pull it straight out before turning off the water tap, to avoid suck-back.
} Need to know safety issues such as handling, processing issues as } above, storeage and disposal of waste. We are assuming that all work } will be done in a fume hood.
As regards general handling, THF is a moderately toxic, highly flammable, clean burning organic solvent. No dreadful horrors associated with it in that respect. The ONE THING you must beware of is (1) distilling it or (2) letting quantities of it evaporate dry. If allowed to stand around (especially if it is not stabilized with hydroquinone or similar antioxidant) it undergoes AUTOXIDATION, which gives rise to organic peroxides. These concentrate in the residue, and are liable to detonate.
It can be distilled, but certain precautions have to be taken. If wanted pure and particle free, purification in a column of something like activated alumina, followed by filtration through a fine membrane filter, could be appropriate. But for your application this may be using a sledgehammer to crack a nut - a good quality reagent straight out of the bottle could well be good enough. But store the bottle in the dark, and don't use the very last dregs.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Researcher in the area of "Electron microscopy in plant research". Reference number 1716/98-4599.
At the Electron Microscopy Unit, Department of Plant Breeding Research, The Swedish University of Agricultural Sciences, Sval=F6v/Alnarp, Sweden. The EM Unit will be a common resource for the university's departments in Southern Sweden.
The candidate must have a doctor's degree, primarily not earlier than five years ago. Experience of work with plant material is a merit. Of importance are the scientific, pedagogic, adminstrative and other capacities of relevance for this occupation. Of importance also is the capacity to inform about scientific research in a popular way.=20
The appointment is first for two years, followed by another period of two years and the possibility for a further prolongation. The salary is individually determined (at least 20 000 Swedish crowns a month, before taxes). Women are engouraged to apply.
Together with the application, a short account on the applicant's scientific and pedagogic activities should be given. The application marked with the above-mentioned reference number, together with a certified C.V., merit and publication lists, and other documents (including scientific and pedagogic works) should be sent in two copies so that they reach the "Registrator, SLU, Box 7070, S-750 07 Uppsala, Sweden" at the latest on June 5, 1998.
For further information contact professor Waheeb Heneen, tel +46 418 667064 or fax +46 418 667081.
} } To meet requirements for the Joint Comission, I have been asked by the } laboratory manager to come up with Age Specific Competencies as part of our } Position Description Format for each employee. To my knowledge, there are } no age specifiic competencies for histotechs as they have no patient } contact. } } Can anyone help me with this? } Thanks!! } } Kathy Pelton -Henrion } Supervisor of Histology } SUNY HSC at Syracuse
Well, knowing college students, you might not want to let anyone under the age of 21 near the absolute ethanol.
Charlie Ginsburg NCC
_________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
One minor error in the previous discussion: The Mag-i-cal standard is produced by John McCaffrey and not South Bay Technology; the latter, like at least half a dozen other vendors (includ. ProSciTech) are suppliers.
The other consideration when choosing a standard is its efficacy. Some standards are suitable for several calibrations, but most users are mostly, or only concerned with magnification calibration. Probably all "standards" sold for TEM or SEM calibration are sufficiently accurate when properly used.
It is difficult to calibrate a TEM with a greater accuracy than 5% +/- and an SEM is a little worse. If you really "go to town", one could calibrate a TEM possibly with some certainty to 3% accuracy. To use this accuracy, all relevant conditions, especially objective lens current, Z and hysteresis would have to be very accurately reproduced. In SEM especially, specimen tilt and topography undermine highest accuracy.
The Point: A simple grating replica I expect is accurate to better than 1% over a few spaces, the best standards may be, say 4x more accurate. With the instruments almost impossible to usefully calibrate to 3%, the additional accuracy of the standard is likely to only give a false sense of accomplishment. The limitations are reading where the standard's lines start and finish and to reproduce instrument conditions when making measurements. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
At 06:57 PM 5/11/98 EDT, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Very often when hangars are used, the problem turns out to be too much agitation of the negatives during processing. I don't know what your "cloudiness" looks like, but agitation problems often show up as darker edges on negatives, leaving lighter edges on prints. This is caused by increased turbulence of the developer as it flows around the hangers, causing in turn greater development of the film areas near the points of contact.
Quite often, this problem shows up differently depending on who does the processing, because people usually develop their own agitation patterns (a lot like life?). Some folks really like to swish the negatives around, while others move them just enough to ensure that fresh chemicals contact the film on a once-a-minute or so basis.
It's hard to say if this is your problem, but you might try varying your agitation routine to see if it helps. Kodak used to recommend lifting the hangars and tipping first to one side and then another for a few seconds, once per minute, then letting the hangar remain still until the next cycle comes around.
Hope this helps.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Thanks to everyone who responded to my question about lab policies regarding protocol development. The many replies were extremely informative and I will make a summary available to those who requested it as soon as I can get to it. Replies ranged from "The investigator should do as much as possible." to "We know the EM stuff, so we should develop the protocols." Most of the responses, as I suspected, ranged along the middle ground.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
I quite agree with Randy Tindall's assessment of the problem, see my additional comments below:
} } Don Gantz wrote:
} } We have gradually aquired a cloud around the outer edge of our negatives } } processed in an Arkay developer. The problem does not occur in trays or } when } } processing a single hanger. When you process three hangers all nine } negatives } } are cloudy. } } } } We cleaned the developer tank with old fixer and ferricyanide, the hangers } and } } fixer tank with 5% nitric acid and scrubbing. We made up new solution. And } } the problem continues. } } } } We cannot seem to locate Omega-Arkay. Any advice or location of Arkay is } } appreciated.
} Randy Tindall wrote:
} Very often when hangars are used, the problem turns out to be too much } agitation of the negatives during processing. I don't know what your } "cloudiness" looks like, but agitation problems often show up as darker } edges on negatives, leaving lighter edges on prints. This is caused by } increased turbulence of the developer as it flows around the hangers, } causing in turn greater development of the film areas near the points of } contact.
Exactly. I went through this some years ago. Too much agitation creates uneven turbulent flo near the edges of the hangers, resulting in overdevelopment there. (Gib)
} Quite often, this problem shows up differently depending on who does the } processing, because people usually develop their own agitation patterns (a } lot like life?). Some folks really like to swish the negatives around, } while others move them just enough to ensure that fresh chemicals contact } the film on a once-a-minute or so basis.
I develop Kodak TEM negs (*emulsion #4489) in full strength D-19 for 3.0 minutes. I agitate up and down,lifting the rack completely out of developer on the upstroke, twice when I first insert film into developer, then JUST ONCE every 45 seconds thereafter until time is up. I get even development doing this. (Gib)
SNIP!
} Randy Tindall } Electron Microscope Laboratory } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } } rtindell-at-nmsu (work) } nrtindall-at-zianet.com (home) }
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
Thanks to everyone who volunteered ideas about how to help a user in my lab stain many grids. I received nearly 30 replies and each one added greatly to my ability to help my user. I now feel like the world's expert on multiple grid staining.
Isn't this list idea wonderful? Three cheers to all of you and a special thanks to Nestor for his efforts to keep it working.
Thanks again.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
can I get some sources for a cathodeluminescence detector for a Hitachi s2700 SEM?
thanks
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
} } Hello, } } We are trying to get a 3-dimensional picture of a plant meristem using our Confocal Microscope. The plant that we are studying is very hairy and seems to be very resinous or waxy. We are having trouble getting fixatives into the tissue and also clearing the tissue. The stains we have tried are Saffranin and Fast green. These both fluoresce but neither is getting into the tissue very well. I have read that Coryphosphine O is a good plant cell wall stain that fluoresces, but I have not had any success in tracking down a supplier. If anyone can help me with a supplier for this or has done confocal work on this sought of sample I would very much appreciate some advice. I have done work on maize meristems that work quite well with propidium iodide. This only stains DNA/RNA etc so is not suitable for our purpose with the other tissue as we need to see cell walls. } Please, can anyone Help? } } Thankyou } } Liz }
We use a Nitrogen Burst system and this ensures an even agitation of the developer. The Nitrogen flow rate can be set to a level where the right amount of turbulence occurs around the negative. This is an operator independent solution.
We did have a problem with patchy negatives a number of years ago. This was identified as an Ilford negative problem. We changed to Agfa and have had no problems since.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland.
Tel: 353-1-6081820 Fax: 353-1-6770438
----------------------------------------------------------------------. } } I quite agree with Randy Tindall's assessment of the problem, see my additional } comments below: } } } } Don Gantz wrote: } } } } We have gradually aquired a cloud around the outer edge of our negatives } } } processed in an Arkay developer. The problem does not occur in trays or } } when } } } processing a single hanger. When you process three hangers all nine } } negatives } } } are cloudy. } } } } } } We cleaned the developer tank with old fixer and ferricyanide, the hangers } } and } } } fixer tank with 5% nitric acid and scrubbing. We made up new solution. And } } } the problem continues. } } } } } } We cannot seem to locate Omega-Arkay. Any advice or location of Arkay is } } } appreciated. } } } } Randy Tindall wrote: } } } Very often when hangars are used, the problem turns out to be too much } } agitation of the negatives during processing. I don't know what your } } "cloudiness" looks like, but agitation problems often show up as darker } } edges on negatives, leaving lighter edges on prints. This is caused by } } increased turbulence of the developer as it flows around the hangers, } } causing in turn greater development of the film areas near the points of } } contact. } } Exactly. I went through this some years ago. Too much agitation creates uneven } turbulent flo near the edges of the hangers, resulting in overdevelopment there. } (Gib) } } } } Quite often, this problem shows up differently depending on who does the } } processing, because people usually develop their own agitation patterns (a } } lot like life?). Some folks really like to swish the negatives around, } } while others move them just enough to ensure that fresh chemicals contact } } the film on a once-a-minute or so basis. } } I develop Kodak TEM negs (*emulsion #4489) in full strength D-19 for 3.0 } minutes. I agitate up and down,lifting the rack completely out of developer on } the upstroke, twice when I first insert film into developer, then JUST ONCE } every 45 seconds thereafter until time is up. I get even development doing this. } (Gib) } } SNIP! } } } Randy Tindall } } Electron Microscope Laboratory } } Box 3EML } } New Mexico State University } } Las Cruces, NM 88003 } } } } rtindell-at-nmsu (work) } } nrtindall-at-zianet.com (home) } } } } } } Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor } Electron Optical Facility, University of Minnesota, Dept. Plant Pathology } 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 } 612-625-9728 FAX, giba-at-puccini.crl.umn.edu } } "Theory and practice are the same in theory, but different in practice." } }
Does anyone still use this technique or seen this discussed elsewhere? I'm also interested in finding any non-biological applications. Jeff Day,( "JD") Mesquite, Texas WA5EKH-at-JUNO.COM
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
Researcher in the area of "Electron microscopy in plant research". Reference number 1716/98-4599.
At the Electron Microscopy Unit, Department of Plant Breeding Research, The Swedish University of Agricultural Sciences, Sval=F6v/Alnarp, Sweden. The EM Unit will be a common resource for the university's departments in Southern Sweden.
The candidate must have a doctor's degree, primarily not earlier than five years ago. Experience of work with plant material is a merit. Of importance are the scientific, pedagogic, adminstrative and other capacities of relevance for this occupation. Of importance also is the capacity to inform about scientific research in a popular way.=20
The appointment is first for two years, followed by another period of two years and the possibility for a further prolongation. The salary is individually determined (at least 20 000 Swedish crowns a month, before taxes). Women are encouraged to apply.
Together with the application, a short account on the applicant's scientific and pedagogic activities should be given. The application marked with the above-mentioned reference number, together with a certified C.V., merit and publication lists, and other documents (including scientific and pedagogic works) should be sent in two copies so that they reach the "Registrator, SLU, Box 7070, S-750 07 Uppsala, Sweden" at the latest on June 5, 1998.
For further information contact professor Waheeb Heneen, tel +46 418 667064 or fax +46 418 667081.
Hello All, I must disagree with Jim Darley on the calibration accuracy you can get from a TEM. We routinely get about 1% or better accuracy in magnification of our JEOL 120CX, and use it several times a week to measure III-V layer thicknesses for laser structures. All you need to do is: 1) make sure the sample is at eucentric height (although mag is very slow changing with this, you can be quite a way off and there's no difference in the image); 2) take all lenses in the imaging system to their maximum setting and back just before taking the picture (all the knobs end up in the about same place each time - we quickly noticed the reference batteries dying this way, since the standard position began to move); 3) only use the central part (~3cm diameter) of the image (distortions of up to 5% can occur close to the edges). When the calibration errors are combined with measurement errors (typically 0.05 mm with a 10x lupe and 'ruler' with 0.1mm lines - if the interfaces are sharp!) our measurements usually have errors of 1-2%. This has been checked with 'blind' tests of standard samples over several years. We have found diffraction grating replica standards to be of little use since a) it is hard to make sure the grating is not tilted in the microscope, and b) there are variations in the spacings of up to 5%.
Richard Beanland
GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK
_____________________________________________________________________________ } One minor error in the previous discussion: The Mag-i-cal standard is } produced by John McCaffrey and not South Bay Technology; the latter, like at } least half a dozen other vendors (includ. ProSciTech) are suppliers. } } The other consideration when choosing a standard is its efficacy. Some } standards are suitable for several calibrations, but most users are mostly, } or only concerned with magnification calibration. Probably all "standards" } sold for TEM or SEM calibration are sufficiently accurate when properly } used. } } It is difficult to calibrate a TEM with a greater accuracy than 5% +/- and } an SEM is a little worse. If you really "go to town", one could calibrate a } TEM possibly with some certainty to 3% accuracy. To use this accuracy, all } relevant conditions, especially objective lens current, Z and hysteresis } would have to be very accurately reproduced. In SEM especially, specimen } tilt and topography undermine highest accuracy. } } The Point: A simple grating replica I expect is accurate to better than 1% } over a few spaces, the best standards may be, say 4x more accurate. With the } instruments almost impossible to usefully calibrate to 3%, the additional } accuracy of the standard is likely to only give a false sense of } accomplishment. The limitations are reading where the standard's lines start } and finish and to reproduce instrument conditions when making measurements. } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } Great microscopy catalogue, 500 Links, MSDS, User Notes } **************************** www.proscitech.com.au *****
If you are planning to attend the MMMS Materials meeting to be held at the University of Illinois-Chicago on May 22, please RSVP the organizer, Dr. Nigel Browning at: BROWNING-at-UIC.EDU
Vendors: If you are planning to attend and wish to have table space available for your literature, please E-mail the Corporate Laison, Chris Wadelton at: CWADELTON-at-AOL.COM.
Please reply directly to the above addresses, not to the listserver.
Regards, Chris Wadelton Corporate Laison KLA-Tencor/Amray division
Greetings, Liz Nickless wrote: } I have read that Coryphosphine O is a good plant cell } wall stain that fluoresces, but I have not had any success in tracking down } a supplier. Coriphosphine O is (or at least used to be) sold by Polysciences. (If you don't have a contact number for them I can probably find their fax for you.) Yes, it stains plant cell walls nicely, and is excitation and emission characteristics are close enough to rhodamine for all practical purposes. I have no idea about its "penetrability", etc. Good luck, Tobias Baskin
I would be interested in some thoughts on digital cameras and digital systems in general. We are trying to upgrade our metallurgical failure analysis lab to incorporate digital imaging and hopefully eliminate the need for Polaroid and 35mm film. We haven't purchased anything yet.
We have three input devices to connect to the computer workstation: 1) Polaroid MP4 macro stand, 2) Leica stereo microscope and 3) Leco metallograph. I was hoping use the Kodak DCS 410 w/ Nikon N90 camera body on the macro stand and interchange the DMC2000 on the two microscopes. I am not set on these choices and I'd appreciate any comments you have on these cameras or comparable ones.
The other problem I have is which printer to use. The reports we generate are typically 10-15 pp of B&W text followed by 50-75 pp of B&W and color images. We distribute 12-15 copies of each report. I've seen some encouraging results from the Epson Stylus 800, but I'm concerned about its speed and ability to handle the volume. What about color laser printers? Are they worth the 5-6K? I have some demos printed on the Tektronics Phaser 560 - the color prints looked decent, but the B&W prints were not as good as the HP LaserJet 5 at 600 dpi. I looked at dye subs also, but the cost/page seemed high. Thermal wax and solid ink didn't have the image quality we were looking for.
You might look into the Pixera for a lower cost camers ($1200) We have recently installed one on our optical microscope and find it very good for most applications. It is not adequate for low light applications.
We have also just purchased an Espon Stylus Photo printer. It was under $300. I think and produces outstanding, near photo quality prints. Much better than a 1200 DPI laser printer. It is slow but it is cheap. We are ordering several more for our EM work. When it comes time for publications quality prints we use our dye-sub printer
Greg Erdos ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Assistant Director, The Biotechnology Program PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
I have been asked by a colleague to look into performing SEM on a hair sample from a patient with a rare disease called pili bifurcati. The problem is that the sample with the pathology of interest has been mounted on a glass slide and coverslipped with the non-aqueous mounting media, permount.
There are some free hairs for analysis, however none of them show the pathology of interest. Can the sample on the slide be placed in alcohol to remove the permount, rinsed and dried for SEM prep? Or does anyone have any suggestions on how to salvage this mounted hair for analysis, if possible? Any suggestions would be appreciated (I am a TEM person by trade, sorry if Ive left out any SEM info needed for this question)
Thanks much,
Susan Danielson, MS Neuromuscular Laboratory Coordinator Medical College of Wisconsin / Froedtert Hospital
I wonder does anyone has an experience in operating with both BioFilter and Leo912 microscopes especially in spectroscopic modes. These microscopes have appeared few years ago on the market, however there is still very few publications indicating the using one of them. I would like to know the difference in microscopes’ performance when operated in a spectroscopic mode and coupled with a dark-field mode, in particular. Is there any significant difference in image quality depending on GIF or Omega filters especially for phosphorus elemental analysis. Or does the quality and resolution of the acquired images strongly depend on slow-scan CCD cameras used either by built-in GIF100 or bottom-attached Leo 912 from various manufactures.. I would be deeply appreciated for any information related to the posted questions.
The only age-specific competency standard that comes to mind for our = Histology is that we paraffin block only tonsils of children under 12 = unless otherwise indicated (clinical, or at time of gross). Full work-up = for tonsils of children 12 y and up.
You may want to post on the Histonet to see what response you get there.
Regards,
Peter O. Steele, Ph.D., All Children's Hospital St. Petersburg, Fl.
Normal Disclaimer Applies: Views expressed are my own and not necessarily = that of All Children's Hospital etc., etc.
} } } "Kathleen Pelton-Henrion (Kathleen Pelton-Henrion)" {PELTONK-at-mailbox.hs= csyr.edu} 6:17:08 PM 5/11/98 } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html=20=
To meet requirements for the Joint Comission, I have been asked by the laboratory manager to come up with Age Specific Competencies as part of = our Position Description Format for each employee. To my knowledge, there are no age specifiic competencies for histotechs as they have no patient contact.
Can anyone help me with this? Thanks!!
Kathy Pelton -Henrion Supervisor of Histology SUNY HSC at Syracuse
Depending on the nature of the pathology, about which I know nothing, the answer is "yes". You'll have to remove the Permount by soaking in xylene, not alcohol (leastways I could never soak off Permount with alcohol, only xylene), but after the hairs have been removed from the slide, and given a gentle swirl in a change or three of xylene to remove any residual Permount, take them down to EtOH through a 1:2 1:1 2:1 EtOH:xylene series (1:1 only might work, but I'm paranoid), give them a change or three in 100% EtOH, then dry for SEM.
I assume any damage that xylene would do to the hairs was already done before mounting them in the Permount. Or were they just stuck in Permount and coverslipped? If this is the case, because of possible damage by xylene, I would pop off the coverslip and Permount with hairs with dry ice or liquid nitrogen (as for freeing resin sections that have been re-embedded). Then try carefully chipping away the Permount from the hairs while submerged in liquid nitrogen.
Phil } } I have been asked by a colleague to look into performing SEM on a hair } sample from a patient with a rare disease called pili bifurcati. The } problem is that the sample with the pathology of interest has been } mounted on a glass slide and coverslipped with the non-aqueous mounting } media, permount. } } There are some free hairs for analysis, however none of them show the } pathology of interest. Can the sample on the slide be placed in alcohol } to remove the permount, rinsed and dried for SEM prep? Or does anyone } have any suggestions on how to salvage this mounted hair for analysis, } if possible? Any suggestions would be appreciated (I am a TEM person by } trade, sorry if Ive left out any SEM info needed for this question) } } Thanks much, } } Susan Danielson, MS } Neuromuscular Laboratory Coordinator } Medical College of Wisconsin / Froedtert Hospital }
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 5037 Station A Champaign, IL 61825-5037 (217) 355-1143 oshel-at-shout.net or poshel-at-hotmail.com ***** looking for a job *****
Very nicely stated. The most important thing with respect to setting up the microscope and this includes SEM's as well is to set the microscope up the same every time. I've been surprised at the number of experienced SEM uses that don't know what Eucentric is.
I've used a calibration sample in the past that was a calibrated square grating replica with calibrated polystyrene balls. The grids weren't exactly square, but the balls did have a very consistent size to them. However, when I used both sizes in the mag range that both were usable, there was a significant difference in the mag found between them. When I got to PPG, I had to use the Mag standards that were here because of our QS9000 certification. I bought the Mag-i-cal sample and redid the mags. The agreed fairly well, but the Mag-i-cal sample is much easier to use and seems to be very consistent. Plus, it gives me all the calibrations that I need in one sample.
But again, the most important thing is to set up the microscope the same way each time. For imaging, make the sample eucentric and focus. For critical cases, all the lenses should be brought to the same setting from the same way to avoid hysteresis as Richard says. For diffraction, there are two ways to set up the Intermediate 1 lens (i.e. diffraction focus knob): 1) after the image is eucentric and focus and then switched over to diffraction mode, fully spread the condenser lens to make the incident beam parallel and focus the transmitted spot to the smallest spot with the diffraction focus. This method does not take the Back focal plane image of the lens for the image plane of the first intermediate lens. or 2) after the image is eucentric and focus and then switched over to diffraction mode, condense the beam so that it is at crossover for convergent beam mode and then either focus HOLZ lines if they are visible or focus on the shadow image of the condenser aperture. This method does grab the back focal plane for the image plane of the first Intermediate lens.
I've measured layer thicknesses in III-V systems and have gotten similar accuracies as Richard. In addition, the values that I get are consistent with other measurements on the same samples using XRD, ellipsometry, and growth rates.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Hello All, I must disagree with Jim Darley on the calibration accuracy you can get from a TEM. We routinely get about 1% or better accuracy in magnification of our JEOL 120CX, and use it several times a week to measure III-V layer thicknesses for laser structures. All you need to do is: 1) make sure the sample is at eucentric height (although mag is very slow changing with this, you can be quite a way off and there's no difference in the image); 2) take all lenses in the imaging system to their maximum setting and back just before taking the picture (all the knobs end up in the about same place each time - we quickly noticed the reference batteries dying this way, since the standard position began to move); 3) only use the central part (~3cm diameter) of the image (distortions of up to 5% can occur close to the edges). When the calibration errors are combined with measurement errors (typically 0.05 mm with a 10x lupe and 'ruler' with 0.1mm lines - if the interfaces are sharp!) our measurements usually have errors of 1-2%. This has been checked with 'blind' tests of standard samples over several years. We have found diffraction grating replica standards to be of little use since a) it is hard to make sure the grating is not tilted in the microscope, and b) there are variations in the spacings of up to 5%.
Richard Beanland
GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK
________________________________________________________________________ _____ } One minor error in the previous discussion: The Mag-i-cal standard is } produced by John McCaffrey and not South Bay Technology; the latter, like at } least half a dozen other vendors (includ. ProSciTech) are suppliers. } } The other consideration when choosing a standard is its efficacy. Some } standards are suitable for several calibrations, but most users are mostly, } or only concerned with magnification calibration. Probably all "standards" } sold for TEM or SEM calibration are sufficiently accurate when properly } used. } } It is difficult to calibrate a TEM with a greater accuracy than 5% +/- and } an SEM is a little worse. If you really "go to town", one could calibrate a } TEM possibly with some certainty to 3% accuracy. To use this accuracy, all } relevant conditions, especially objective lens current, Z and hysteresis } would have to be very accurately reproduced. In SEM especially, specimen } tilt and topography undermine highest accuracy. } } The Point: A simple grating replica I expect is accurate to better than 1% } over a few spaces, the best standards may be, say 4x more accurate. With the } instruments almost impossible to usefully calibrate to 3%, the additional } accuracy of the standard is likely to only give a false sense of } accomplishment. The limitations are reading where the standard's lines start } and finish and to reproduce instrument conditions when making measurements. } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } Great microscopy catalogue, 500 Links, MSDS, User Notes } **************************** www.proscitech.com.au *****
I have four associates looking for a JEOL preferably an 840 series. I also have three associates looking for an "in lens" field emission SEM such as the Hitachi S-5000.
I wonder does anyone has an experience in operating with both BioFilter and Leo912 microscopes especially in spectroscopic modes. These microscopes have appeared few years ago on the market, however there is still very few publications indicating the using one of them. I would like to know the difference in microscopes=ED performance when operated in a spectroscopic mode and coupled with a dark-field mode, in particular. Is there any significant difference in image quality depending on GIF or Omega filters especially for phosphorus elemental analysis. Or does the quality and resolution of the acquired images strongly depend on slow-scan CCD cameras used either by built-in GIF100 or bottom-attached Leo 912 from various manufactures.. I would be deeply appreciated for any information related to the posted questions.
I'm interested in comments from users of the Pixera Professional camera such as Greg ). We are considering purchasing one because of the low cost/high resolution. I am a little worried about the small chip size, lack of interpolation, and X4 resolution enhancement. They don't really explain how this works, except to say that they are real pixels. Does it work ? I would be very interested if any users would mail a sample image materials preferably ) so that I could look at the image quality and try print-outs here.
Thanks Kirk for relaying my request for a local supplier of the Pixera. I have been in touch with Grant johnson ( ISS ) in the UK.
On the subject of printing we have had a Xerox4900 colour laser for about four years and do not find it good enough for producing images. It is great for solid blocks of colour but does not reproduce the colours very well and colour images proved very difficult to output correctly. I am sure that things have improved in today's colour lasers but at a price. We recently bought a couple of Epson Stylus 800's and now use them for all colour work and high quality B&W prints. Using their photo paper it produces near photo quality prints which are accepted by our customers in place of conventional photos. The speed ( & media costs ) is a major problem though and we only use it for high quality prints, with most of our throughput on a HP Laserjet 600 dpi. We recently found a low cost supplier of photo quality paper, but the Epson paper feed can leave puncture tracks which shows up on some printouts.
Colin
Colin Reid, Electron Microcope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. -----Original Message-----
Colin: Our online has quite a lot of info on digital photography in general and Pixera's triple pass advantage in particular. A number of images are also at that site; they are not bad but note the internet is limited to 72 ppi and its useless to reproduce images larger than common screen sizes. Also tiff files suffer a bit when reduced into JPEG's. ProSciTech markets Pixera and I am happy to declare that interest. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} } Hi, } } I'm interested in comments from users of the Pixera Professional camera } such as Greg ). We are considering purchasing one because of the low } cost/high resolution. I am a little worried about the small chip size, } lack of interpolation, and X4 resolution enhancement. They don't really } explain how this works, except to say that they are real pixels. Does it } work cut ....
} Colin Reid, } Electron Microcope Unit, } Trinity College Dublin, } Dublin 2, } Rep. of Ireland.
Hello- We are trying to examine cracks in etched nylon by SEM. We wish to look at them on edge to see the depth and morphology. We are concerned with embedding them in epoxy, as there may be insufficient contrast between the nylon and the filler. Does anyone have any experience or ideas that they might share on this subject? Thank You.
I'm sure many noted Scott Walck's comment on how "surprised" he was that so many users (TEM and SEM) - when asked, I guess, - did not not know what eucentric means. I am one of those users who doesn't know, but after Scott's message I took a little time to "TRY" and educate myself about this "topic". To that end, I turned to Goldstein et. al (1992), but to no avail. (eucentric is not in the index). Next I tried "SEM: A User's Manual for Materials Science" by Gabriel - again to no avail. (not in the index). Next "Electron Optical Systems for Microscopy, Microanalysis and Microlithography" edited by Hren et. al - you got it, to no avail! Finally, as a last resort I turned to my Webster's Collegiate Dictionary for at least a general definition (figuring it was an old term from light-optical miroscopy)....nothing!!
} From this, I can only assume it is either a very new issue, perhaps unique to TEMs and some modern SEMs, or, that it is a very obscure issue, but one that needs attention because of its emergence as a topic on this list. SO, can soemone out there shed some light on this subject or direct me to a reference on it?
Thanks, in advance, for those of you who take the time to respond. Also, yes, I will attempt a synopsis of the answers which will be circulated at a later date.
Winton Cornell
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
A eucentric stage on either an SEM or a TEM is one in which there is an adjustment to change the height of the sample relative to the pivot point of the tilt axis. If a sample is at the eucentric height, then it is at the same height physically with respect to the objective lens and hence the focal plane of the lens when the sample is focussed will be at exactly the same place. You can tell when the sample is not eucentric when you tilt the sample and it sweeps across the screen. When you go through the eucentric point, the sample will sweep the other way. At the eucentric point, it will not move, but just change the apparent width of features in a direction perpendicular to the tilt axis.
Once a sample is eucentric and then focused, subsequent areas of that sample or new samples can be made eucentric by not touching the focus knob and focussing with the eucentric height adjustment.
I think that most of the newer SEMs have eucentric stages. Some older ones do not. some have you adjust the height of the sample in the sample holder prior to inserting them into the microscope. All TEMs have eucentric stages.
Your manual for the microscope should have the eucentric adjustment described in it.
I'm sure many noted Scott Walck's comment on how "surprised" he was that so many users (TEM and SEM) - when asked, I guess, - did not not know what eucentric means. I am one of those users who doesn't know, but after Scott's message I took a little time to "TRY" and educate myself about this "topic". To that end, I turned to Goldstein et. al (1992), but to no avail. (eucentric is not in the index). Next I tried "SEM: A User's Manual for Materials Science" by Gabriel - again to no avail. (not in the index). Next "Electron Optical Systems for Microscopy, Microanalysis and Microlithography" edited by Hren et. al - you got it, to no avail! Finally, as a last resort I turned to my Webster's Collegiate Dictionary for at least a general definition (figuring it was an old term from light-optical miroscopy)....nothing!!
} From this, I can only assume it is either a very new issue, perhaps unique to TEMs and some modern SEMs, or, that it is a very obscure issue, but one that needs attention because of its emergence as a topic on this list. SO, can soemone out there shed some light on this subject or direct me to a reference on it?
Thanks, in advance, for those of you who take the time to respond. Also, yes, I will attempt a synopsis of the answers which will be circulated at a later date.
Winton Cornell
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
In our free publication "Invitation to the SEM World" we devote a half page to this subject wherein it is defined as "A specimen stage designed so that the specimen tilt axis agrees with the observation point, is called the eucentric stage. Its' feature is that the field of view is not shifted and the focus is not largely changed by specimen tilting." There are also diagrams and sample micrographs showing both eucentric and non-eucentric stages.
This is a non-commercial, educational booklet. If anyone would like a copy (or copies) please email me your address and I would be happy to provide it.
We also have quicktime or avi movies showing this function which are part of our multimedia CD. The CD is commercial of course but if there were a lot of interest we could cut that part out and include it in the download section of our web site as a strictly educational offering. Again let me know.
Stephen Hamilton Tel: 978-536-2270 JEOL USA, Inc. Fax: 978-536-2205 11 Dearborn Road Email: hamilton-at-jeol.com Peabody, MA 01960 WWW: http://www.jeol.com
} -----Original Message----- } From: Winton Cornell [mailto:wcornell-at-centum.utulsa.edu] } Sent: Thursday, May 14, 1998 10:32 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Eucentric - is there a good overview out there? } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Listers: } } I'm sure many noted Scott Walck's comment on how "surprised" he } was that so } many users (TEM and SEM) - when asked, I guess, - did not not know what } eucentric means. I am one of those users who doesn't know, but after } Scott's message I took a little time to "TRY" and educate myself } about this } "topic". To that end, I turned to Goldstein et. al (1992), but to } no avail. } (eucentric is not in the index). Next I tried "SEM: A User's Manual for } Materials Science" by Gabriel - again to no avail. (not in the } index). Next } "Electron Optical Systems for Microscopy, Microanalysis and } Microlithography" edited by Hren et. al - you got it, to no } avail! Finally, } as a last resort I turned to my Webster's Collegiate Dictionary for at } least a general definition (figuring it was an old term from light-optical } miroscopy)....nothing!! } } } From this, I can only assume it is either a very new issue, } perhaps unique } to TEMs and some modern SEMs, or, that it is a very obscure issue, but one } that needs attention because of its emergence as a topic on this list. SO, } can soemone out there shed some light on this subject or direct me to a } reference on it? } } Thanks, in advance, for those of you who take the time to respond. Also, } yes, I will attempt a synopsis of the answers which will be } circulated at a } later date. } } Winton Cornell } } } } } Dr. Winton Cornell } Senior Research Associate & Supervisor, Microanalysis Laboratory } Department of Geosciences } The University of Tulsa } 600 South College } Tulsa, OK 74104-3189 } } phone: 918-631-3248 } fax: 918-631-2091 } e-mail: wcornell-at-centum.utulsa.edu } }
Winton asks: } } I'm sure many noted Scott Walck's comment on how "surprised" } he was that so } many users (TEM and SEM) - when asked, I guess, - did not } not know what eucentric means. ... }
I am aware of it being an expensive option for an SEM, but probably is very useful for a TEM. It isn't new, and I believe it allows for tilting and rotation without the object "swinging" out of the field of view.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
Eucentric is a purely geometric term and thus can apply to any situation which involves geometry (not just TEMs!). Specifically, "eu-" means "good" or "true": the word means a system which has multiple axis of rotation, which intersect at a point (i.e. a true centre of rotation). Consider a matched set of goniometers (such as described in any optics catalog): these are said to be designed to "ensure that they rotate about a common point". That is "eucentric". In microscopy applications it ensures that you stay focussed at the same point as you tilt, twist and rotate the sample (always a useful feature).
does anyone know what an appropriate oil alternative is for a JEOL35C DP (DP4E) that has a tag that says "use LION-S" or is this type pretty common?
thx brian
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax) "Be well, do good work, and keep in touch"
We prepared some iodine-doped epoxy for mounting coal. The iodine raised the atomic number enough that we got plenty of BSE contrast, but it would also give SE contrast.
The process involves dissolving iodoform in the epoxy resin above room temperature and then treating the doped resin much the same as regular epoxy resin except for a little more care in handling. Iodoform is not good for you.
I forget who first describe this procedure back in the early to mid 80s, but could dig up a reference in needed.
At 07:04 AM 5/14/98 -0700, you wrote: } } Hello- } We are trying to examine cracks in etched nylon by SEM. } We wish to look at them on edge to see the depth and morphology. We are } concerned with embedding } them in epoxy, as there may be insufficient contrast between the nylon } and the filler. } Does anyone have any experience or ideas that they might share on this } subject? } Thank You. } } } Charles Hubka } AlliedSignal Inc.
We routinely mount powders(metal or ceramic) in 1.25 inch diameter mounts of Bakelite or epoxy, then examine the polished surface in the light microscope and then the SEM. Sometimes we will even send the mounts to another facility for microprobe analysis. I would like to know if anyone has a good method for locating specific areas of the mount, similar to the finder grids for TEM, so that we could easily locate multiple areas as we switch between instruments.
F. Scott Miller Electron Microscopy Lab smiller-at-umr.edu University of Missouri-Rolla 223 McNutt Hall voice: 573 341 4727 Rolla, MO 65409 USA fax: 573 341 6934
There will be an opening for an SEM operator in Tucson Arizona sometime in the coming months. This position is to replace me. Interested parties should contact me directly via e-mail or phone. A background in electrochemistry is preferred. Thank you.
Stephanie Wind McCray Process Chemist & SEM Lab Mngr. Moltech Corp. 9000 S Rita Rd, Bldg 61 Tucson, AZ 85747 520-799-7631 (office/lab) wind-at-moltech.com
To assist those with the meaning of the term Eucentric here follows the = answers.
A eucentric stage allows the operator of am EM to tilt the sample = without loosing the area of interest . In most stages, as you tilt the = stage the area of interest will move out of site. This means that as you = tilt the sample, you need to correct the X and Y stage drives, to keep = the same area of interest in view. The eucentric stage allows you to tilt and still see the same area at = the new angle without adjusting the X or Y drives. However most Eucentric stages are only Eucentric at a certain working = distance. As you move away from this point the stage acts more like a = normal stage.
ISI / Topcon SEM's have offered this as standard for about15 years. Most = other manufacturers can also offer this type of stage as an option.=20 We have a LEO S440 user with a eucentric stage. However they never need = to tilt their samples, so I can't say how well it works.
I hope this helps.
Cheers. Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
-----Original Message-----
Listers:
I'm sure many noted Scott Walck's comment on how "surprised" he was that = so many users (TEM and SEM) - when asked, I guess, - did not not know what eucentric means. I am one of those users who doesn't know, but after Scott's message I took a little time to "TRY" and educate myself about = this "topic". To that end, I turned to Goldstein et. al (1992), but to no = avail. (eucentric is not in the index). Next I tried "SEM: A User's Manual for Materials Science" by Gabriel - again to no avail. (not in the index). = Next "Electron Optical Systems for Microscopy, Microanalysis and Microlithography" edited by Hren et. al - you got it, to no avail! = Finally, as a last resort I turned to my Webster's Collegiate Dictionary for at least a general definition (figuring it was an old term from = light-optical miroscopy)....nothing!!
} From this, I can only assume it is either a very new issue, perhaps = unique to TEMs and some modern SEMs, or, that it is a very obscure issue, but = one that needs attention because of its emergence as a topic on this list. = SO, can soemone out there shed some light on this subject or direct me to a reference on it?
Thanks, in advance, for those of you who take the time to respond. Also, yes, I will attempt a synopsis of the answers which will be circulated = at a later date.
Winton Cornell
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
Actually, it's not new or obscure, even if it has apparently been omitted by some book authors! You will find it in D.B. Williams & C.B. Carter, Transmission Electron Microscopy, Plenum Press, 1996. Hope this helps.
Gill Bond Dept Materials & Met. Eng. New Mexico Tech
On Thu, 14 May 1998, Winton Cornell wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Listers: } } I'm sure many noted Scott Walck's comment on how "surprised" he was that so } many users (TEM and SEM) - when asked, I guess, - did not not know what } eucentric means. I am one of those users who doesn't know, but after } Scott's message I took a little time to "TRY" and educate myself about this } "topic". To that end, I turned to Goldstein et. al (1992), but to no avail. } (eucentric is not in the index). Next I tried "SEM: A User's Manual for } Materials Science" by Gabriel - again to no avail. (not in the index). Next } "Electron Optical Systems for Microscopy, Microanalysis and } Microlithography" edited by Hren et. al - you got it, to no avail! Finally, } as a last resort I turned to my Webster's Collegiate Dictionary for at } least a general definition (figuring it was an old term from light-optical } miroscopy)....nothing!! } } } From this, I can only assume it is either a very new issue, perhaps unique } to TEMs and some modern SEMs, or, that it is a very obscure issue, but one } that needs attention because of its emergence as a topic on this list. SO, } can soemone out there shed some light on this subject or direct me to a } reference on it? } } Thanks, in advance, for those of you who take the time to respond. Also, } yes, I will attempt a synopsis of the answers which will be circulated at a } later date. } } Winton Cornell } } } } } Dr. Winton Cornell } Senior Research Associate & Supervisor, Microanalysis Laboratory } Department of Geosciences } The University of Tulsa } 600 South College } Tulsa, OK 74104-3189 } } phone: 918-631-3248 } fax: 918-631-2091 } e-mail: wcornell-at-centum.utulsa.edu } }
} } does anyone know what an appropriate oil alternative is for a JEOL35C DP } (DP4E) that has a tag that says "use LION-S" or is this type pretty common? } thx } brian
Brian
A few years ago I went thru the same exercise, then discovered that Lion S is available from JEOL, for about half the price of Santovac. I don't know what it is, though, does anyone else out there? I replaced the Fomblin oil (which I had inherited in my old JEOL probe) with Lion S and was immediately rewarded with a better vacuum (mind you, the Fomblin was pretty old) and faster pumping. I think this says something about pumps being optimised for particular oils.
On this subject, though, is anyone out there successfully using Santovac in an Edwards 306 coater? Did you have to change the heater?
Ritchie
} Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Brian McIntyre wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } hi all- } } does anyone know what an appropriate oil alternative is for a JEOL35C DP } (DP4E) that has a tag that says "use LION-S" or is this type pretty common? } } thx } brian } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627 } } 716-275-3058 } 716-244-4936(fax) } "Be well, do good work, and keep in touch"
Lion-S is pretty common. I purchase mine from Topcon 800.538-6850. Their part number is 210 136.
Brian McIntyre wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } hi all- } } does anyone know what an appropriate oil alternative is for a JEOL35C DP } (DP4E) that has a tag that says "use LION-S" or is this type pretty common? } } thx } brian } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627 } } 716-275-3058 } 716-244-4936(fax) } "Be well, do good work, and keep in touch" Hello Brian, I would recommend Kurt J. Leskers "Diffoil 20" or Inland 20. I believe these oils to be actually superior to the origional in that they will provide you with less backstreaming and at the same time work well for your purpose. Inland Vacuum is Churchville, N.Y. Phone 800/962-8099 & Kurt Lesker Co. is in Pennsylvania Phone 800/245-1656.
Other companies make similar products and my company is in no way affiliated with the makers of these products. Either of these companies offer nice catalogs and excellent service.
Alexander Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. Number 499, Post Office Box 19400 Austin, Texas
Electron Microscope Repair Phone: 512/282-5507 FAX 512/280-0702
We have learned more about the Jeol Jem100U unit (40 years old, w/ goniometer stage, good condition, w/ vacuum evaporator, no longer serviced by Jeol) since I first inquired about the value of this scope that was offered as a donation to our high school.
It's likely that this scope may not be worth much. If any potential user of this instrument feels otherwise, please contact me with your expression of interest in its acquisition.
Thanks.
Tom DeVries tomdevrie-at-aol.com 206-463-9171 x314
Philips produced a side-entry eucentric stage for their EM 200 TEM in the late 1960s. The point being that you could vary the position of the rod holding the specimen so the major tilt axis could be positioned to lie in the front focal plane of the objective and also intersect the axis of the lens. Or be centred on the axis. Then any tilting of the specimen around the major tilt axis did not result in disappearance of the field of view. But it was only "eucentric" for the major axis and tilting on the secondary axis in a double tilt holder could not be so compensated. Siemens built a completely eucentric stage in the 70s but the price was more than the market could take.
Most side entry tilt stages are direct derivatives of the original Philips design and are only partly eucentric. Philips now produce a computer controlled fully eucentric stage in which both axes of tilt can be brought to intersect the principal lens axis in the focal plane.
SEM eucentric stages also aim to be able to locate the specimen on the major tilt axis so tilting does not result in loss of the field of view.
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
I know of a used Hitachi S-7080 for sale. This is a fully automated semiconductor wafer analyzing tool. Vacc=0.1-3.0kv handles 6-8 inch wafers more info available on request
Allen Johnson donjuan-at-juno.com
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
Brian McIntyre wrote: =============================================== does anyone know what an appropriate oil alternative is for a JEOL35C DP (DP4E) that has a tag that says "use LION-S" or is this type pretty common? =============================================== The firm CVC Products, Inc. tradenamed their dioctylsebacate Octoil®-S some years ago, it was around as early as about 1970, perhaps even earlier. When we took delivery of our JSM-U3 SEM in early 1970, the recommended diffusion pump fluid by JEOL USA, Inc. was Octoil-S. We don't know if this was the very first dioctylsebacate D. P. fluid, but it was the first that came to my attention.
Over the years, others have offered their own brand of dioctylsebacate XXXXX -S, Lion™ -S being one of them. Another one that was visible was Invoil™ - S (Inland Oil Company). I do not know the owner of the trade name Lyon-S, however since JEOL has been the source of the Lyon-S fluid it might very well be their trade name. Bottles labelled Invoil-S started appearing on the scene in the mid-1970's, and the JEOL service engineers started using those bottles instead of bottles marked Octoil-S. The same engineers at a later date started using bottles labelled Lyon-S. Without proof, of course, it was always our belief that generically at least all three brands of pump fluids were equivalent. At least we never saw any change in performance of our SEM with the conversion of one brand to another brand.
Over the years, our own SPI packaged bottles of Octoil-S have been used without incident in all SEMs that had been previously using any of of the other XXXX-S dioctylsebacate diffusion pump fluids. The use of dioctylsebacate in this application has been on the decline since Santovac® 5 came onto the scene. Although it is more expensive, it has far greater resistance to decomposition when someone does something not quite so smart, or there is a failure of the vacuum system somewhere, and air gets into the hot diffusion pump fluid.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I have just learned that NIST has released the entire source code for Desktop Spectrum Analyzer (DTSA) which was Developed by Chuck Fiori, Carol Swyt-Thomas, and Bob Myklebust as FREEWARE. I realize that many of you may not be interested in the code itself, but I thought you might like to know it is available at:
http://micro.nist.gov/dtsa/dtsa.html
Here is a brief synopsis taken from the NIST Site
Desktop Spectrum Analyzer (DTSA)
Developed by Chuck Fiori, Carol Swyt-Thomas, and Bob Myklebust
A MACINTOSH ONLY! based x-ray spectral analysis and manipulation software package.
The NIST/NIH Desktop Spectrum Analyzer generates, interprets and analyzes x-ray spectra from specimens under electron bombardment. This remarkable software/database package simulates the experimental environment and emulates specimen properties to generate spectra reflecting the relevant physics, chemistry and statistics of a real world application. DTSA incorporates many widely accepted x-ray data analysis procedures including those developed over many years at the National Institute of Standards and Technology (NIST) in Gaithersburg, MD and the National Institutes of Health (NIH) in Bethesda, MD.
Hi all, On the subject of DP oils, may I just warn users to just check with the = manufacturer of the DP pump as to what oil is recommended. In the past a local technician decided to use the Dow Corning 702 , 705 = and 704 oils, which according to the supplier would work much better. After a six month period on each system where he had changed the DP oil, = we found Si peaks in all the spectra they collected on their ED systems. Dow corning oils are Si based and this oil impregnates the Be window and = then gives the Si peaks. Lion S seems to be a Japanese DP oil as we find that most Topcon Jeol = and Hitachi systems use this. Santovac is only used by the Topcon DS 130 Sem and most Edwards system. It is very expensive but also very resilient.
Cheers Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
I am pleased to announce the website of the EUREM 12
12th EUROPEAN CONGRESS ON ELECTRON MICROSCOPY to be held in Brno, Czech Republic July 9 - 14, 2000
at http://www.eurem2000.isibrno.cz/
Best regards,
Petr Schauer +---------------------------------------------------------------------+ | Dr. Petr Schauer, Secretary of the tel.: (+420 5) 41514313 | | Czechoslovak Soc. for Electron Microscopy fax : (+420 5) 41514404 | | ACADEMY OF SCIENCES OF THE CZECH REPUBLIC (+420 5) 41514402 | | INSTITUTE OF SCIENTIFIC INSTRUMENTS e-mail: petr-at-isibrno.cz | | Kralovopolska 147, CZ-612 64 Brno csem-at-isibrno.cz | | Czech Republic www: http://www.isibrno.cz/csem/ | +---------------------------------------------------------------------+
To all who like to observe stereo images: a TEM answer and a SEM question:
The basic principle of eucentricity is the same for SEM and TEM, but in practice there are some very strong differences:
On our TEM (Philips 301) it is easy to adjust a grid carrying a heavy metal shadowed carbon replica to the eucentric height: then one can tilt a few degrees either side about the main axis of the goniometer stage to get images viewable under an appropriate binocular viewer. However, if the angle of application of the heavy metal shadow (the azimuth, not the altitude) differs significantly from the tilt axis, then the contrast will increase when rocked one way, and decrease on the other.
The SEM (Philips 515) is different. If one has the stage in normal (not dropped) position and adjusts Z so that the Filament Working Distance to sample is 12 mm, then one is at the eucentric. But tilting is about the other axis, and gives the impression of an aircraft looking diagonally down at the surface. So one will not get proper stereo pairs simply by tilting. One could apply rotation and tilting, but that requires some clever spherical trigonometry - does anybody know the answer? Or is there a mechanical solution?
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
} } I would appreciate hearing from anyone who has experience using } Tetrahydrofuran (THF) for dissolution of PVC filter membranes and } redeposition of particules remaining in the THF solution onto silver } membrane filters. This is done by vacuum filtration. } } What types of vacuum pumps can safely be used for this purpose and can } THF be sonicated. } } Need to know safety issues such as handling, processing issues as } above, storeage and disposal of waste. We are assuming that all work } will be done in a fume hood. } John -
There are a few OSHA procedures for preparing air samples collected on PVC filters for analysis by x-ray diffraction. If I recall correctly, both the crystalline free silica and the vanadium pentoxide methods suggest using THF as a solvent for dissolving the filter. My understanding is that not all of the PVC filters available on the market can be prepared using this method.
Mike Rose at OSHA's Salt Lake City Technical Center has done a great deal of work in this area. You may want to consider contacting him to discuss this in greater detail. I think he can be reached at 801-487-0267 or you may be able to place an inquiry at their web site {http://www.osha-slc.gov} .
Keith Rickabaugh Manager, Materials and Particle Characterization {krickabaugh-at-rjlg.com}
RJ Lee Group, Inc. 350 Hochberg Road Pittsburgh, PA 15146 ph: 724-325-1776 {www.rjlg.com}
The choice of DP oils depends on numerous tradeoffs. Price, chemical stability, resistance to oxidation or cracking, vacuum performance, and how it behaves in an electron beam can be relevant for SEM/TEM/Microprobe usage. LION-S is one of many brands of dioctylsebacate oil, a low cost and readily available diffusion pump oil. Santovac-5 is one of several brands of polyphenyl ether DP oils. Fomblin is one of several brands of perfluorinated DP oils. Numerous silicone based DP oils are common. Silicone oils are very rugged and very forgiving of air exposure/ oxidation/cracking and are very chemically stable. Unfortunately, silicone oils form insulating silicon-based deposits when exposed to an electron beam and contaminates the column, apertures, and system and is rarely recommended for use in SEM/TEM/Microprobes for this reason. Dioctylsebacate pumps well and is very reasonably priced but is not very forgiving of air leaks or vacuum accidents. It also readily forms organic "carbon deposits" or polymers when exposed to electron beams and is often replaced by other pump fluids to minimize this. Polyphenyl ethers and perfluorinated oils are considerably higher priced but offer significantly higher resistance to oxidation/air leaks/ cracking/vacuum accidents. Electron beam interactions don't deposit as much organics with polyphenyl ethers as they do with dioctylsebacate. Perfluorinated DP fluids deposit even less when exposed to electron beams, and offer the advantage of the deposits being relatively conducting and don't affect column performance as much as they build up. Also perfluorinated fluids are even more resistant to degradation or air exposure. However, the heater power requirements in a DP pump with perfluorinated fluids is about 50% power (70% voltage) compared to most other oils, and usually requires the use of an adjustable autotransformer to get proper pumping. Not reducing the power will increase backstreaming and degrade ultimate vacuum. I have been using perfluorinated DP fluids for many years on a number of systems successfully.
These comments are my own opinions and do not reflect the endorsement of my employer or of any of the products mentioned.
Garber, Charles A. wrote: } =20 } -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } =20 } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } =20 } Brian McIntyre wrote: } =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D } does anyone know what an appropriate oil alternative is for a JEOL35C D= P } (DP4E) that has a tag that says "use LION-S" or is this type pretty com= mon? } =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D } The firm CVC Products, Inc. tradenamed their dioctylsebacate Octoil=AE-= S some } years ago, it was around as early as about 1970, perhaps even earlier. = When } we took delivery of our JSM-U3 SEM in early 1970, the recommended diffu= sion } pump fluid by JEOL USA, Inc. was Octoil-S. We don't know if this was = the } very first dioctylsebacate D. P. fluid, but it was the first that came = to my } attention. } =20 } Over the years, others have offered their own brand of dioctylsebacate = XXXXX } -S, Lion=99 -S being one of them. Another one that was visible was In= voil=99 - } S (Inland Oil Company). I do not know the owner of the trade name Lyon= -S, } however since JEOL has been the source of the Lyon-S fluid it might ver= y } well be their trade name. Bottles labelled Invoil-S started appearing = on } the scene in the mid-1970's, and the JEOL service engineers started usi= ng } those bottles instead of bottles marked Octoil-S. The same engineers = at a } later date started using bottles labelled Lyon-S. Without proof, of co= urse, } it was always our belief that generically at least all three brands of = pump } fluids were equivalent. At least we never saw any change in performan= ce of } our SEM with the conversion of one brand to another brand. } =20 } Over the years, our own SPI packaged bottles of Octoil-S have been used } without incident in all SEMs that had been previously using any of of t= he } other XXXX-S dioctylsebacate diffusion pump fluids. The use of } dioctylsebacate in this application has been on the decline since Santo= vac=AE } 5 came onto the scene. Although it is more expensive, it has far great= er } resistance to decomposition when someone does something not quite so sm= art, } or there is a failure of the vacuum system somewhere, and air gets into= the } hot diffusion pump fluid. } =20 } Disclaimer: SPI offers Octoil-S dioctylsebacate and Santovac 5 polyph= enyhl } ether diffusion pump fluids. } =20 } Chuck } =20 } =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } =20 } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D Santovac 5 uses a higher DP temperature than the Octoil S and, therefore, is not a direct repalcement for Octoil S or Invoil. I have interchanged the invoil with the lion S and have consistently achieved better pressure with the lion s. I am told that both are similar in chemistry but are not similar in results.
I purchase lion s from topcon although JEOL I am sure has stock. Topcon's direct number is 201. 261-9450.
Dear Robert, You wrote: (snip) } then one is at the eucentric. But tilting is about the } other axis, and gives the impression of an aircraft looking diagonally } down at the surface. So one will not get proper stereo pairs simply by } tilting. One could apply rotation and tilting, but that requires some } clever spherical trigonometry - does anybody know the answer? Or is there } a mechanical solution?
Yes, when you do a stereo pair, the scan must be rotated 90 degrees to put the tilt axis in line with the horizonal, since horizontal is how you will view the stereo. I always think of a line, representing the direction of tilt, being drawn straight through both images as they are viewed. In fact, it is easier to correct this if the stage is not eucentric, as you use the sweep of one part of the image to get your raster rotation right on the horizontal. Hope this helps.
Regards, Mary Mager
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
Dear Brian, The LION-S" is a Japanese DP oil and I have successfully used Santovac-5 as a replacement in all my Hitachi microscopes. I believe most other quality DP oils will also work. You wrote: } hi all- } } does anyone know what an appropriate oil alternative is for a JEOL35C DP } (DP4E) that has a tag that says "use LION-S" or is this type pretty common? } } thx } brian } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
If you have one or know of a Philips-made single tilt heating holder which fits a Philips 430, I would be interested in finding out details for purchase or trade. Please contact me directly.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Is everyone REALLY SURE that Lion-S oil is similar in composition and properties to Octoil-S, and the other "-S" oils??
The question of the nature of Lion-S DP oil came up on this circuit almost exactly a year ago. Like most others, I had always assumed that it was similar to the other "-S" oils on the market such as Octoil-S, Invoil-S, Diffoil-S, etc, in being the synthetic compound di-(2-ethyl-hexyl)-'S'ebacate. HOWEVER, there remained a bit of uncertainty in my mind, and so I pestered Mike Kersker of JEOL long enough and hard enough to finally get him to check with sources within his company, and he came back with the information that Lion-S oil is an alkylnaphthylene compound. He also supplied the address of the Lion Oil Company (1-22-2 Yokoami, Suymida, Tokyo, Japan; Tel: 03-3621-6684) - but the company did not respond when I wrote a letter asking for specs on their oil.
The only other diffusion pump oil I could find that might be similar to Lion-S, if indeed it is an alkylnaphthalene compound, is Alcatel-200 fluid (obviously marketed by Alcatel). This fluid is reported to be eicosyl naphthalene, which is indeed an alkynaphthalene compound. The properties of this fluid are summarized on pp. 182-3 of my book 'Vacuum Methods in Electron Microscopy' (see: http://www.cccbi.chester.pa.us/spi/catalog/books/book48.html or http://www.bookshop.co.uk/portland/). As shown there, the Alcatel-200 fluid has a substantially lower vapour pressure (below 10-7 Pa at 20=B0C) than Octoil-S (about 3x10-6 Pa at 20=B0C). That is, the vapour pressure characteristics of Alcatel-200 resemble those of Santovac-5 and DC-705 much more closely than those of Octoil-S. Thus, IF(!!) Lion-S resembles Alcatel-200 more closely than di-(2-ethyl-hexyl)-Sebacate, then replacing it with one of the di-(2-ethyl-hexyl)-Sebacate fluids could lead to a degradation in the performance of your pump. Instead, Santovac-5 or the Alcatel-200 fluid would be a better replacement.
Since this matter of replacing Lion-S oil in diffusion pumps seems to keep coming up fairly frequently, I will write to the Lion Oil Co. again, and try to get reliable information from them. In the meantime, if any of you have such info, I would appreciate knowing about it.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
I have completed the design and construction of my latest attachemnt for the HRTEM at Oak Ridge Nat'l Lab, and will be going down there to check it out over the week of May 17th, and then going on to South Carolina to visit friends there. I expect to be back in the office on Monday, June 1st.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Dear Listers, A client has asked me to size graphite particles treated with approx. 0.2% picric acid using the SEM. The material has been handled quite a bit but the MSDS fire and explosion hazard and reactivity make me wary of proceeding. The samples (in dried powder form) were deposited onto double sticky C tape in a fume hood. I would appreciate advice from chemists, previous experience, precautions and suggestions re: accv. Rosemary
#################################################### Rosemary Walsh Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:rw9-at-psu.edu ####################################################
Now that 111-Trichloroethane (Inhibisol, Genklene etc.), and Trichlorotrifluoroethane (Arklone) are no longer being manufactured, does anyone know of any other solvents available which are suitable for cleaning EM vacuum systems and internal column parts?
Over the years I have used Acetone, Diethyl Ether and 40/60 Petroleum Ether, but all these carry a fire risk, and Acetone softens paintwork.
Any information on available alternatives would be appreciated.
Bob Phillips microservis-at-dial.pipex.com http://dspace.dial.pipex.com/microservis/
} Now that 111-Trichloroethane (Inhibisol, Genklene etc.), and } Trichlorotrifluoroethane (Arklone) are no longer being manufactured, } does anyone know of any other solvents available which are suitable } for cleaning EM vacuum systems and internal column parts? } } Over the years I have used Acetone, Diethyl Ether and 40/60 Petroleum } Ether, but all these carry a fire risk, and Acetone softens paintwork. } } Any information on available alternatives would be appreciated. } We have just completed our annual dis-assembly & cleaning of the HVEM lens column, and we use a detergent (Alconox) solution, several rinses with distilled water and ethanol. For the more recalcitrant gunk we have used acetone, a freon, Inhibisol, or chloroform. We have a very good air-circulating system--the whole Wadsworth Center is at negative pressure with respect to the outside--and the air is completely changed in about 20 minutes. We have not, therefore, had any fire problems. Ethanol is very clean, and it is our solvent of choice (assuming it dis- solves whatever is on the scope parts); and it makes a good finishing wash even when another solvent must be used first. Yours, Bill Tivol
Can anyone supply me with reference(s) to consult when planning an EM laboratory? I am particularly concerned about isolation from vibration and climate control. Thank you.
Does anyone have any experience with the newest Olympus inverted scope? For brightfield or fluorescence...pro, anti, indifferent, whatever? I was asked and haven't a clue - haven't even seen one - but I said I'd ask around.
Yes, I obviously know about the Email Spam's from the Mass Market Emailer.. They are relayed from different computers over different network connections every time.
Just so you know the Email address of the Listerserver is posted on a number WWW sites around the world and shall we call them "creative individuals" have written Internet search robots to cruise the WWW and look specifically for Email addresses. These addresses are then added to mailing lists. Which are sold or used. That is likely the method that these spammers have found us.
I am looking at different ways to get rid of this junk mail, all of which involve either alot of work, or compromises in the way this system runs.
For now, it is certainly true that the mail frequently does not contain text in the subject line, and it's orginator is usually an alphanumeric string which is not a "name" and has not yet been a subscriber. The hosts unfortunately are not always the same site so I cannot block the mail by simply putting a denial of service block on a particuliar DNS.
One of the options I am considering is writing an Email filter that will automatic trash any Email message that has a blank subject line. So please, make sure you use a subject line at all times.
Also as long as I have your attention, we are still getting the occassional posting of messages for Microscopy at the administrative address. I manually forward all of these to the correct address but as a reminder they are:
Messages to the Group: Microscopy-at-MSA.Microscopy.Com Administrative Messages: Listserver-at-MSA.Microscopy.Com
Subscribe/Unsubscribe/Help should go to Listserver-at-... Posting of Questions to the Group should go to Microscopy-at-...
You can also use the WWW Site address documented at the top of this message to Subscribe/Unsubscribe.
I have a question for those of you with JEOL 2010F microscopes (or anyone else with a view on the matter). We want to know if an uninteruptable power supply (UPS) is necessary to maintain the gun operation in the event of a power failure.
We know of at least one lab who have installed a UPS on their new 2010F, but we have received advice from a colleague that it shouldn't be necessary. What is the general concensus?
If a UPS was to be employed, what components of the microscope operation should it maintain and how should it be configured to operate?
Any feedback would be greatly appreciated. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
At 18:43 15/05/98 EDT, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The reference to use is "Design of the Electron Microscope Laboratory" 1975
Ronald H Alderson
Vol 4 in "Practical Methods in Electron Microscopy" Audrey M. Glauert, Editor,
Library of Congress # 73-94298
North-Holland ISBN 0 7204 4259 1
/Elsevier ISBN 0 444 10807 6
I have used it in designing 3 laboratories. If the architects / engineers had only read the copies I gave them, all would have gone much better!
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Dear All I am trying to find a supplier of uranyl formate to use as a negative stain. I would prefer a UK supplier for convenience but would like to hear from anyone who could supply.
Many thanks
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
I'm not sure if it would help in this particular situation, but some of you may be interested in a web site that provides translations. The address is http://babelfish.altavista.digital.com/cgi-bin/translate? You type in the text, or the URL of a web page that needs to be translated, and it will provide translations from French, German, Italian, Portugese or Spanish into English, and vice versa. I don't know how good it is at handling technical material.
Leslie Eibest Zoology Dept., Box 90325 Duke University Durham, NC 27708 USA (919) 684-2547 leibest-at-duke.edu
In many countries the solvents you mention are either banned or simply no= t available. As a result the service engineer has to improvise. A very go= od "solvent" for the less waxy cleaning agents is good old hot soapy water! = I clean with Silvo or the wadding metal polish Duraglit which wash away nicely, using cotton buds when an internal wipe is required..
I have used this technique around the world there seems to be only one problem. When you take the parts to the sink to clean them make sure you=
return with the same number of parts. I guess I am the world's expert on=
taking apart sink systems that have not been apart since they were installed a million years ago. Not a nice job but often the only way to find that missing fixed aperture. =
Provided you use a suitable cleaning media (like those mentioned above) I=
find dilute ammonia (5 to 10% solution in water) a solvent absolutely ide= al for stainless steel parts. It is very good for a second reason, it is al= so a solvent for tungsten.
Steve Chapman
Senior Consultant Ed.MA. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
George, You might want to try Alta Vista's translation service at: http://babelfish.altavista.digital.com/cgi-bin/translate? As far as I know it is a free service and the few times that I have tried it I have gotten credible results, although it probably won't handle the scientific terms well. Greg
George Theodossiou wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi all, } } I've got a paper but its in French and I need the english translation } if it exists. CAN ANYONE HELP ME!! } } The paper is: } } Title: Cerium Pretreatments To Prevent Filiform Corrosion Of Painted } Aluminium Alloys } } Author: V.Poulain J.-P Petitjean } } Source: Mater. Sci. Forum (Switzerland), Materials Science Forum, } vol. 217-222, pt.3, p. 1641-8 } } ISSN: 0255-5476 } G. Theodossiou } Dept Applied Physics } RMIT } Email: George-at-bunyip.ph.rmit.edu.au } ph:+61 3 9925 3394 } fax:+61 3 9925 5290
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Hi there, I dont know if this is the right place to look for responses, but; Does anyone have a good reference that should be looked at to learn how to pull your own micropipettes/needles and cell holders that you would use for micromanipulation work. Specifically the removal of single cell contents from plants? although I'm sure those used for mammalian cells would work. Or is the experience that it's just better to buy them (Im in Australia)?
Any hints would be greatly appreciated.
(im doing this for a colleague, so I dont know the specs of the equipment, but I could find out. Yes we do have a microforge and other pieces of equipment, we just haven't the experience to use them properly.)
Hi there, I dont know if this is the right place to look for responses, but; Does anyone have a good reference that should be looked at to learn how o pull your own micropipettes/needles and cell holders that you would use for micromanipulation work. Specifically the removal of single cell contents from plants? although I'm sure those used for mammalian cells would work. Or is the experience that it's just better to buy them (Im in Australia)?
Any hints would be greatly appreciated.
(im doing this for a colleague, so I dont know the specs of the equipment, but I could find out. Yes we do have a microforge and other pieces of equipment, we just haven't the experience to use them properly.)
I am posting this question for a student. Please reply directly to her if you can help with her problem.
Her name is Nazima Shahnoor and her e-mail address is:
naz-at-csd.uwm.edu
I have muscle tissue fixed in formaldehyde that was washed with ethyl alcohol then stored in 70% EtOH at room temperature for about 1 year. I have successfully used Phosphotungstic acid/hemotoxalin (PTAH) for staining A bands in formaldehyde-fixed tissue (not stored in EtOH), however for this tissue the PTAH protocol is giving poor results. Is there an alternative protocol for observing A bands in this tissue, or is there any way to "rescue" this tissue so that PTAH will work?
Thank you for any help.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
Dear all, =20 =20 I'm after a list of atomic positions for the unit cell of an=20 intermetallic phase called CHI (x) often encountered in stainless=20 steels. I am not sure about the space group (I-43m?) and would be=20 grateful if someone could help me on this. =20 F.
One thing that has been missed from the discussion is the effect of sample height on X-ray counts if you are doing EDX. At least for TEMs, the EDX quantification assumes that one is at the eucentric height, errors can be introduced if one is not at the eucentric height.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Does anyone know of a source for 12mm diameter and 11mm X22mm (approximately) glass #1.5 coverslips? TIA Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine Houston, TX
Responding to the message of {v03102802b185fa6074a3-at-[147.188.152.52]} from Ian MacLaren {I.MacLaren-at-BHAM.AC.UK} : } } One thing that has been missed from the discussion is the effect of sample } height on X-ray counts if you are doing EDX. At least for TEMs, the EDX } quantification assumes that one is at the eucentric height, errors can be } introduced if one is not at the eucentric height. } I am not sure that this is strictly true. The position of the sample for EDX analysis should be the intersection of the electron beam with the axis of the spectrometer. Only in this case is the X-ray take-off angle (and therefore all the quantitative calculations) correct. Deviations from this height obviously matter more with low take-off angle detectors.
Whilst it is convenient to have this height coincident with the eucentric height it is not essential.
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
Hi Liz, sorry I didn't reply earlier to this, I was out of town. Did anyone suggest that you try calcofluor white to stain cell walls? It fluoresces pale blue upon excitation below 400 nm. I can provide you with staining protocol if you desire. Cheers, John
} } We are trying to get a 3-dimensional picture of a plant meristem using our } Confocal Microscope. The plant that we are studying is very hairy and seems } to be very resinous or waxy. We are having trouble getting fixatives into } the tissue and also clearing the tissue. The stains we have tried are } Saffranin and Fast green. These both fluoresce but neither is getting into } the tissue very well. I have read that Coryphosphine O is a good plant cell } wall stain that fluoresces, but I have not had any success in tracking down } a supplier. If anyone can help me with a supplier for this or has done } confocal work on this sought of sample I would very much appreciate some } advice. I have done work on maize meristems that work quite well with } propidium iodide. This only stains DNA/RNA etc so is not suitable for our } purpose with the other tissue as we need to see cell walls. } } Please, can anyone Help? } } } } Thankyou } } } } Liz } }
================= C. John Runions, Ph.D Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
I would like to know how medical EM labs handle specimens that potentially harbor Creutzfeldt-Jakob or other prion mediated diseases. We get very few calls to work up such specimens, but when we do, there is an awkward time deciding how to handle it, or whether to handle it at all. There is enough material in the literature on their resistance to fixatives and other agents that render bacterial and viral specimens safe to handle that people are quite reluctant to work with such specimens. I would like to hear from medical EM labs telling me whether or not you accept such specimens; and if you do, what special procedures do you use?
I understand that this might be a subject that has already been exhausted on the list server. However I have been off-line a while. If anyone remembers the consensus, I would appreciate a review. .........There HAS been a consensus on this server, hasn't there?
Joiner Cartwright, Jr., Ph.D. Director, Electron Microscopy Lab Department of Pathology
The Chi phase is also known as the A15 phase or alpha-Mn phase. This is according to a monograph by A. Sinha (Prog. Mater. Sci vol 15, p. 79 (1972)). The atom positions are given there.
Incidentally the work cited is a fine reference on the group of compounds known as topologically close-packed compounds.
Wharton
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu http://www.numis.nwu.edu/internet/Staff/wharton/wharton.html
On Mon, 18 May 1998 frank.sarrazit-at-avestasheffield.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear all, } } } I'm after a list of atomic positions for the unit cell of an } intermetallic phase called CHI (x) often encountered in stainless } steels. I am not sure about the space group (I-43m?) and would be } grateful if someone could help me on this. } } F. }
any other additions to this discussion are welcome
At 12:52 PM 5/18/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Hi, I only speak English, so I don't know how accurate Babelfish is, however I= have spent some time amusing myself translating quotes to other languages and back to English. For example:
"There is a theory which states that if ever anyone discovers exactly what= the Universe is for and why it is here, it will instantly disappear and be rep= laced by something even more bizarrely inexplicable. There is another theory which states that this has already happened."
Translated to Portugese becomes: "H=E1 uma teoria que indique que se sempre qualquer um descobrirem exatamente o que o universo =E9 para e porque ele est=E1 aqui, ele desaparecer=E3o e ser=E3o substitu=EDdos imediatamente por algo mesmo mai= s bizarrely inexplicable. H=E1 uma outra teoria que indique que esta tem acontecido j=E1 "
Translated back to English is: "He has a theory that he indicates that if always any one to discover accu= rately what the universe is for and because it is here, it will disappear and wil= l be substituted by something exactly more bizarrely immediately inexplicable. He has one another theory that he indicates that this has happened already= . "
Pearson: Handbook of Lattice Spacings and Structures of Metals, also gives atom positions for alpha-Mn (and many other) phases.
Larry Thomas Washington State University email: thomas-at-mme.wsu.edu or Larry.Thomas-at-pnl.gov
---------- From: Wharton Sinkler Sent: Monday, May 18, 1998 6:01 PM To: frank.sarrazit-at-avestasheffield.com Cc: Microscopy-at-Sparc5.Microscopy.Com Subject: Re: CHI-Phase
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Frank,
The Chi phase is also known as the A15 phase or alpha-Mn phase. This is
according to a monograph by A. Sinha (Prog. Mater. Sci vol 15, p. 79 (1972)). The atom positions are given there.
Incidentally the work cited is a fine reference on the group of compounds known as topologically close-packed compounds.
Wharton
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu http://www.numis.nwu.edu/internet/Staff/wharton/wharton.html
On Mon, 18 May 1998 frank.sarrazit-at-avestasheffield.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear all, } } } I'm after a list of atomic positions for the unit cell of an } intermetallic phase called CHI (x) often encountered in stainless
} steels. I am not sure about the space group (I-43m?) and would be
} grateful if someone could help me on this. } } F. }
Thankyou to everyone that responded to my query on confocal microscopy of plant meristems. With our Confocal microscope we are restricted to excitation ranges of 488nm, 568nm and 620 so the low range stains will not work on our system. Thanks to the person that gave us the contact for Coryphosphine O ie. Polysciences. I have looked this company up on the internet and now have a contact number. We are also looking at some of the references we were given for other stains. Thanks again.
} } Post-Doctoral Research Associate } } } } } } } } A post-doctoral research associate position is available with the } } Industrial Associates Program in Transmission Electron Microscopy at the } } Center for High Resolution Electron Microscopy at Arizona State University. } } The Industrial Associates Program consists of member companies with an } } interest in advanced transmission electron microscopy. The successful } } candidate will gain experience working on real industrial materials } } problems in an academic setting. This unique perspective provides } } excellent training for individuals interested in expanding and broadening } } their skills. } } } } This position is sponsored by major chemical company and offers } } opportunities to work on a wide range of commercial and model heterogeneous } } and homogeneous catalyst system using the most advanced state-of-the-art } } transmission electron microscopy techniques. The primary focus of this } } research will be the development and application of environmental electron } } microscopy to industrially relevant catalyst systems. Areas of particular } } interest include the study of phase transformations under reaction } } environments, in situ polymerization, mobility and sintering of small metal } } particles and dynamic microstructural changes. } } } } The ideal candidate will have a Ph.D. in material science, material } } physics, solid state chemistry or chemical engineering, with extensive } } experience in catalyst characterization by transmission electron } } microscopy. Experience in the areas of catalyst synthesis, testing and } } characterization is preferred. An ability to interact well with others and } } assist industrial scientists in materials problem solving is essential. } } } } Applicants should submit their resume together and the names of 3 } } referees to: } } } } Dr. Peter A. Crozier } } Industrial Associates Program } } Center for Solid State Science } } Arizona State University } } Tempe, AZ 85287-1704 } } } } Peter A. Crozier } } Industrial Associates Program } Center for Solid State Science } Arizona State University } Tel: 602 965 2934 } Fax: 602 965 9004 } } Website: http://www.asu.edu/clas/csss/IAP/
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
************************************************* * Bill Hardy, President * * American Nuclear Systems, Inc. * * 1010 Commerce Park Dr., Suite G * * Oak Ridge, TN 37830-8026 * * (800) 980-9284 FAX: (423) 482-6253 * * www.qtmsys.com Email: bhardy-at-qtmsys.com * *************************************************
Patricia Glazebrook wrote that the choice between high-end color printers seemed to be between Codonics and the Fujix Pictrography 3000. A good alternative that not many microscopists know about is the Sienna FotoPrint.
The FotoPrint produces true digital photographs for about 25 cents per 8.5 x 11 inch print. This includes all chemicals and media. Printing a 5 x 7 inch print can further reduce costs.
The FotoPrint uses any conventional photographic paper to produce photographic quality 360dpi color prints. Because of this, not only is the cost per print very low, but the print will have the same durability, archivability, and feel of a real photograph, because it is a real photo. Aside from the quality and price advantages, the Sienna FotoPrint is also fast, with the ability to output up to 100 prints per hour.
At facilities that require true photographic quality, extremely low cost, and do the volume to justify the purchase or lease price, the FotoPrint is a very viable alternative. Matt Irwin ElectroImage, Inc. 277 Nothern Blvd. Suite 101 Great Neck, NY 11021 Phone: 516-773-4305 Fax: 516-773-2955 E-mail: sales-at-electroimage.com
I would agree with Melvyn on both points. We have just moved in to our new laboratory and used Audrey's book as the starting basis for our design. The layout has worked perfectly but the execution of the building plan left something to be desired.
We provided the architects with a comprehensive list of the requirements and they ignored every point. In a large building they managed to surround us with the air-conditioning for the building, an IT switch-room, a lift, and to finish off they ran the mains electricity cable along our outside wall we are in the basement ). We have just discovered that the mains water pipe ( metal & under high pressure ) comes in directly over our Leo S440. We have no windows, the air-conditioning doesn't work, the sump provides sewer smells regularly, and we are below sea-level. Apart from that everything is fine !
The point is that the plan is important but getting the architects & builders to complete the construction successfully is another thing altogether especially when you are just a minor part of the floor space in a new building.
Colin
Colin Reid, Electron Microcope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message-----
Melvyn we were lucky. We found out about 50% of the problems and nipped some of them in the bud prior to completion. We managed to get anti-vibration platforms fitted for the microscopes & moved the power cable back from the wall. It must have cost the builders a packet ? I still look up in amazement at the water pipe though. The sucker's above my head, as I type, and he's big !
Our tactic was to send a letter early on describing the planning of the construction as a disaster, and stating quite clearly that we felt we would not be in a position to move in since it was unlikely to pass a site survey. It had an immediate effect and they even employed a consultant to measure the fields during construction. The microscopes are happy, but their needs are different to the operators who need air & light.
Regards,
Colin
Colin Reid, Electron Microcope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message-----
} Can anyone supply me with reference(s) to consult when planning an EM } laboratory? I am particularly concerned about isolation from vibration } and climate control. Thank you. } Paul Grover
Our laboratory used to be in the cellar and once we got the possibility to participate in the planning of the reconstruction of our faculty-building I started very early to draw the floor-plans for our premises. I did not want to go back to the cellar and reserved space for us on the fourth and fifth floors (the supporting structure of the building is cast on-site and very steady).=20 I had many fights with the architects etc. but got most of my wishes done. I have to admit that there were real fights - I had to threat them with= court. After all that fight and almost living in the construction site we got a laboratory which we have been fairly satisfied. Naturally there have been changes afterwards but is there any laboratory without changes during 15 years? If anyone is interested you are wellcome to pay a visit to our laboratory.
Regards, Jouko
Jouko K. M=E4ki, Ph.D., Laboratory Manager University of Turku, Laboratory of Electron Microscopy Kiinamyllynkatu 10 FIN-20520 TURKU FINLAND Tel: +358 (0)2 333 7318 GSM: +358 (0)40 505 2521 FAX: +358 (0)2 333 7380 http://www.utu.fi/med/em/index.html
We're analysing mineral fibres using SEM and would like to know what type of filter or substrate people would recommend. The type we presently use have far too much structure in them - they're not easy on the eye or on the image analysis system! Any recommendations would be most welcome.
In Europe I would recommend Glaswarenfabrik Carl Hecht Gmbh &Co. KG which is located in Rhon (o is umlauted), Germany.
email: hecht-at-swin.baynet.de.
Be careful that is an N at the end of the word swin in the email address.
I won't send telephone numbers as email normally gets a very quick reply and is a LOT cheaper.
Good luck, Azriel Gorski
On Mon, 18 May 1998, Hank Adams wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anyone know of a source for 12mm diameter and 11mm X22mm } (approximately) glass #1.5 coverslips? } TIA } Hank Adams } Cell Biology Integrated Microscopy Core } Baylor College of Medicine } Houston, TX }
Simon - I have enjoyed success on a couple fiber analysis projects using membrane filters. The easiest one to use in my experience is the "mixed cellulose ester" Gelman product - trade named Metricel. Available pore sizes are 0.45 and 0.8 microns. This filter has the additional nice feature of being "black" - actually gray, but fairly dark especially when water wet. This can help with any ancillary light microscopy you might do along with your SEM analysis. The filters are quite stable under the e-beam. The other filter I've used when the acetone solubility of the cellulosics ruled them out is Whatman's nylon membrane filter (cat #7402-004 at 0.2 micron pore size). This filter is thicker and drains more slowly than the cellulosic but it also behaves reasonably (with light Au sputter coating) in the SEM.
Dave Calvert Eastman Chemical Co. P.O. box 1972 Lincoln Street Kingsport, TN 37664 voice: (423) 229-4943 fax: (423) 229-4558 calvert-at-eastman.com
} -----Original Message----- } From: Simon.Dumbill-at-aeat.co.uk [SMTP:Simon.Dumbill-at-aeat.co.uk] } Sent: Tuesday, May 19, 1998 6:44 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: SEM - filters for fibre analysis } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Dear All, } } We're analysing mineral fibres using SEM and would like to know } what } type of filter or substrate people would recommend. The type we } presently use have far too much structure in them - they're not } easy } on the eye or on the image analysis system! Any recommendations } would } be most welcome. } } Thanks in advance } } Simon } } } ###################################################################### } } Dr Simon Dumbill } AEA Technology Tel: +44 1235 434245 } 220, Harwell Fax: +44 1235 435941 } Didcot Email: } Simon.Dumbill-at-aeat.co.uk } Oxfordshire OX11 0RA } UK } }
A brief question: Has anyone out there used hexadecene as a cryoprotectant? Could you advise me on optimal times, and whether it is a permeating or non-permeating agent?
Cryogenically yours
James Wesley-Smith Electron Microscope Unit University of Natal Durban, South Africa
I am interested in hearing comments from owners/users of Philips CM200's on their instrument's reliability.
Our situation is that we received our CM200 (twin lens, new Philips designed HT tank, EDAX XEDS, Gatan GIF) in Oct'96 and resolution was demonstrated using a tungsten filament in Dec'96. We then had the LaB6 filament installed and have had problems ever since.
Problems started with a misadjusted high tension cable at the tank causing arcing and carbon tracking on the insulator, then a SF6 leak from the bottom of the HT tank. After adjusting the cable and replacing the HT tank the HT began arcing in the emmision chamber between the Wehnelt aperture and first anode every 2 minutes to hours. This would leave craters in the Wehnelt aperture. Since March'97 we have had the accelerator replaced three times, the HT cable replaced twice, the HT tank once(sent with HT board set for factory test condition), the emmision chamber replaced once(had cross threaded anode from factory and an aperture left out, after six week wait), and the entire HT chain (HT tank, cable, emmision chamber) replaced as a tested unit from Einhoven, still not fixing the arcing problem.
After moving the instrument to our renovated lab(Jan'98) and we were still having the arcing problem. Then, all of the power supplies feeding the HT circuitry were replaced, version 12 software installed and the arcing problem seems to be fixed.
After reinstalling the XEDS and GIF we have had problems with image cropping due to problems in the lens program requiring replacement of PROM's and digital to analog converters.
We have also had other annoying problems such as; four camera jams(in the dozen times we have been able to use the instrument), numerous critical backing pump(CBP) errors, and a failed 25V lens power supply(twice).
I met with Philips national sales and service managers in Aug'97 and was told that these problems were not typical and that they would be resolved.
Again I would appreciate any comments and experiences you have had with you CM200's.
David R. Hull NASA Lewis Research Center Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
Dear Fellow Subscribers: I am new to the listserver, but I know that this subject has been addressed at considerable length in the past. However, I would be very grateful if someone could point me towards a compilation of the previous discussions concerning the pros and cons of the various TEM negative scanners currently available on the market. Thank you for your kind indulgence. Maureen Barcza EM Supervisor Dept. Pathology SUNY HSC at Syracuse
i was looking for a listserve with discussions on spray/atomization/droplet evaluation of different materials....by any chance, has anyone come across this yet??? any assistance would be appreciated.
Michael Mandanas Particulate Materials Center Penn State University email: mxm67-at-email.psu.edu
Dear Rosemary, You didn't mention if the graphite has been washed after picric acid treatment. Picric acid is very water soluble and may have already been removed. You could try washing some in distilled water. The picric acid is a strong yellow, so if you don't see any colour you should be alright. If you do, maybe you can wash the samples in distilled water before you test them. Picric is no danger if it is wet and its reactivity is generally low if it has not been reacted with metals. You wrote: } } Dear Listers, } A client has asked me to size graphite } particles treated with approx. 0.2% picric acid } using the SEM. The material has been handled } quite a bit but the MSDS fire and explosion hazard } and reactivity make me wary of proceeding. } The samples (in dried powder form) were deposited } onto double sticky C tape in a fume hood. } I would appreciate advice from chemists, previous } experience, precautions and suggestions re: accv. } Rosemary } } #################################################### } Rosemary Walsh } Electron Microscope Facility for the Life Sciences } The Biotechnology Institute for Research and Education } 1 South Frear Lab } University Park, PA 16802 } 814-865-0212 email:rw9-at-psu.edu } #################################################### } Regards and good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
We are a university materials science department looking for ways to make the EM training of our students (both graduate and undergraduate) more efficient. We have a TEM and a couple of SEM's, all from Philips (software controlled), and an old JEOL SEM with EPMA (mostly manually controlled). Currently the training of new users is handled by the chief technician and is based on one-to-one 'hands-on' instruction. However, with the number of users increasing rapidly, this is becoming more and more of a burden on the chief technician. Therefore, we are looking for ways to give the training to more people at the same time and/or to give training without occupying the instrument. Our first idea was to look for instructional software packages, preferably of the multimedia type (CD ROM based). If you have experience with such software or know of information sources about such software, your comments will be greatly appreciated. Other solutions to our training problem may of course also be suggested. Offers from commercial parties (software vendors/EM vendors) are welcome. We are also interested in instructional software on other microscopic techniques (LM, AFM, AEM) and EDS and WDS, but this interest is of a somewhat less urgent nature.
Thank you,
------------------------------------------------------------ dr. ir. Siegfried V.N. Jaecques K.U. Leuven Dept. Metallurgy and Materials Engineering (MTM) de Croylaan 2 B-3001 Leuven BELGIUM phone +32 16 32 1278 (direct) or +32 16 32 1260 (secretary) fax +32 16 32 1991 ------------------------------------------------------------
Dear Bob, Some other cleaners you might consider are: a strong (10 to 20 %) solution of oxalic acid, hot or boiling, will loosen the hard, baked-on brown residue of the beam path on stainless steel parts. I use Wenol paste to polish parts, acetone to remove the Wenol, then clean, not denatured, ethanol for a final rinse, then blow-dry. Another good abrasive cleaner is Zud, which contains oxalic acid and seems to rinse away quite cleanly. You wrote: } Now that 111-Trichloroethane (Inhibisol, Genklene etc.), and } Trichlorotrifluoroethane (Arklone) are no longer being manufactured, } does anyone know of any other solvents available which are suitable } for cleaning EM vacuum systems and internal column parts? } } Over the years I have used Acetone, Diethyl Ether and 40/60 Petroleum } Ether, but all these carry a fire risk, and Acetone softens paintwork. } } Any information on available alternatives would be appreciated. } } Bob Phillips } microservis-at-dial.pipex.com } http://dspace.dial.pipex.com/microservis/
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
An investigator is having a problem with his procedure on corrosive casting of rat kidney. After flushing the kidney, it is infiltrated with methyl methacrylate. After it is harden, the tissue is dissolved with conc. KOH. He is getting crystals after dissolving. Has not had this problem before. Does anyone have an idea what the crystals are and how to prevent them from forming. Thanks in advance for any help.
George Lawton Microscopy and Imaging Service Center UT Southwestern Medical School at Dallas
} Over the years, others have offered their own brand of dioctylsebacate XXXXX } -S, LionOE -S being one of them.
Are you sure of this, or is it an educated guess?
} Another one that was visible was InvoilOE - } S (Inland Oil Company). I do not know the owner of the trade name Lyon-S, } however since JEOL has been the source of the Lyon-S fluid it might very } well be their trade name.
I have a 1-litre can of "Lion-A Diffusion Pump Oil", with "Lion Fat and Oil Co" and some Japanese characters on it, this is probably the source (ignoring the presumed typo "Lyon").
} Bottles labelled Invoil-S started appearing on } the scene in the mid-1970's, and the JEOL service engineers started using } those bottles instead of bottles marked Octoil-S. The same engineers at a } later date started using bottles labelled Lyon-S. Without proof, of course, } it was always our belief that generically at least all three brands of pump } fluids were equivalent. At least we never saw any change in performance of } our SEM with the conversion of one brand to another brand.
Surely someone over the years must have done an analysis on these? If someone would care to mail me a few drops of Octoil-S I'll run an IR on it and on Lion-S (which I have). Or maybe someone from JEOL could enlighten us?
} Over the years, our own SPI packaged bottles of Octoil-S have been used } without incident in all SEMs that had been previously using any of of the } other XXXX-S dioctylsebacate diffusion pump fluids. The use of } dioctylsebacate in this application has been on the decline since Santovac { { } 5 came onto the scene. Although it is more expensive, it has far greater } resistance to decomposition when someone does something not quite so smart, } or there is a failure of the vacuum system somewhere, and air gets into the } hot diffusion pump fluid.
But can you just substitute directly for Lion-S, or do you need to change the heater element?
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
So, I would like to know if anyone has successfully (and without changing heater elements), made the following substitutions:
1 Octoil-S or Santovac instead of the recommended Lion-S in an old (how old?) JEOL diff pump on a SEM, TEM, or EPMA?
2 Octoil-S or Santovac instead of the recommended Dow Corning 704 in an Edwards 306 coater?
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Ritchie Sims asks ... } } So, I would like to know if anyone has successfully (and without } changing heater elements), made the following substitutions: } } 1 Octoil-S or Santovac instead of the recommended Lion-S in an old } (how old?) JEOL diff pump on a SEM, TEM, or EPMA? } } 2 Octoil-S or Santovac instead of the recommended Dow Corning 704 } in an Edwards 306 coater? } } ...
I've successfully replaced an existing DP oil (... not knowing what it was ...) with Santovac 5 ... but with considerable cleaning and rinsing with hexanes. Regarding the heater elements, I think you can still use the same heater elements unless they can't achieve the boiling point for S5, ... but I had one bad experience with Santovac 5's apparently higher boiling point ... that being the higher temperature destroyed a heater support with was made out of a relatively low melting point alloy. Replacing the support with a better metal solved the problem.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
We are interested in whether anyone has done any histology (LM and/or TEM) on the nerve cells types of the lepodoptera (butterfly) brain. If so, what stains did you use (LM) or procedures for TEM? Also, if you have done any SEM, we would be interested in any information you have on preparations or references you may have.Thank you! Sincerely, Barbara Plowman and Maria Mejia
University of the Pacific School of Dentistry Smith and Kettlewell Institute of Visual Research San Francisco 415-929-6692 email: Bplowman-at-uop.edu
} Our first idea was to look for instructional software packages, } preferably of the multimedia type (CD ROM based). } If you have experience with such software or know of information } sources about such software, your comments will be greatly } appreciated.
There are two basic instructional packages out of the University of Western Australia in Perth you should look at:
Virtual SEM and Virtual EDS the person to contact is
Brendan J. Griffin Centre for Microscopy and Microanalysis The University of Western Australia Nedlands, WA, AUSTRALIA 6907 ph 61-8-9380-2739 fax 61-8-9380-1087 Email: bjg-at-cyllene.uwa.edu.au
I believe that a paper is being presented on Virtual EDS at this years Microscopy & Microanalysis '98 Meeting in Atlanta. (http://www.microscopy.com/MSAMeetings/MMMeeting.html) . Virtual EDS was if my memory serves me correctly is a joint project between U. of Sydney and UWA, Perth.
Virtual SEM was presented at the Microscopy & Microanalysis 96 meeting in Minneapolis. You can go to those proceedings to look up details.
There is also a set of EDS CD's by one of the major EDS manufacturers. I believe it is Oxford/Link but am not 100% positive. You should call your local sales rep and find out from them.
Nestor Your Friendly Neighborhood SysOp.
--------------------------------------------------------------------------- I have no financial interest in any of these products, however, it is true that both Brendan and the Oxford people have bought me an occasional pint from time to time. Hmmm... come to think of it I've probably bought them a few too! Oh well so much for disclaimers.....
At 18:23 19/05/98 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Siegfried, I appreciate your problems. I am unsure of the benfits of software packages for instruction in how to use an EM. But there are enthusiasts who have made such programs (e.g. Brendon Griffin, University of West Australia). Would you use a software package to learn how to ride a bicycle? My solution to the time demands for one-on-one instruction at the actual microscope is to use tape cassette (CD?) instruction. Just like the audio guides you get in good museums to the collection of pictures.
You give each student a player and sit them at the microscope and leave them alone with it. Tell them to do everything the tapes say but nothing else; if they have problems, to call you.
The audio cassettes have the benefits of:
They apply to the real machine and the student is sitting in front of it
Both hands and eyes are free to touch and look - no need to refer to notes
No frowning supervisor is there to intimidate the student
The student can rewind the instructions where they rarely care to admit failure to understand to a supervisor
They can borrow the same tape until they really understand
As well as pointing out the location of the controls, directions can be given to do simple actions. Turn on the high tension: observe the deflection of the beam current meter: turn up the filament heat: what do you see on the screen?: and this can be programmed to build confidence rapidly because the student is doing these things on their own.
It takes a really experienced user to make the master tape, but once made, they are good for years.
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
One of the electrolytic polishing solutions used in the Tenupol Apparatus is a mixture of 30% conc. nitric acid and 70% methanol.
Excluding MSDS on each component - which contain some useful info see Methanol "Safe Handling Procedures", what I am searching for is some current practices for the proper storage of this mixture.
Volume = 1000-1500 mL in a winchester
and properly labelled .(*********************)
How are other Users storing this chemical in the Laboratory?
I am trying to obtain an MSDS of this mixture from Sturers - maybe it will take a few days, if available at all.
Dear Friends, Singapore will be the host for the 7th APEM, 26 to 30 June 2000. We have created a website and it will be regularly updated with information. You can register on-line too. The address is: http://www.med.nus.edu.sg/micsoc/7apem {http://www.med.nus.edu.sg/micsoc/7apem}
If you need more details, feel free to e-mail to me or the web editor. Thank you.
There are actually three CD's from Oxford, two on EDS & one on operation of the Isis system. I have seen the CD's in action and they are reasonably good with very good graphics helping to explain the actions of the various parts of the SEM. The second CD is a bit corny however with a "private eye" directing the investigation of an unknown sample. They will probably appeal to students. Oxford had an advert for the CD's in the latest issue of Microscopy & Analysis magazine (UK).
Best wishes,
Colin
Colin Reid, Electron Microcope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message-----
We have a range of training courses available on floppy disk (we often se= ll into areas that do not have CD ROMs), our Portfolio, a kind of correspondence course, as well as our well known "in house " training courses. In the latter we visit your laboratory and train students in groups of up to 4 covering SEM, TEM and EDX in different courses. We cov= er training on any make of instrument.
If you are carrying out your own training the "disk"courses combined with=
our "Protrain Portfolio" will offer not only instruction but a structured=
practical procedure to bring the student through to a good level of competence.
"Portfolio" is available for TEM and SEM with a range of experiments and test specimens through which the student is taken with guide line texts a= nd practical advice. One "Portfolio" may be used over and over again to ensure all students reach the same standard.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
MULTIMEDIA from Protrain=0D =0D We are now, after considerable demand from our clients, making freely ava= ilable the well =0D known Protrain electron microscopy course "slide shows", demonstrations t= hat we have used in =0D our courses for the last eight years.=0D =0D The slide shows, which in the past were pictures only, now include sound = which makes them a =0D complete lecture course that you may run for yourself over and over again= =2E Because we use =0D these courses ourselves we occasionally see a need for better explanation= s and make changes to =0D try to improve the way we pass on information, these updates will also be= made available to =0D users of the M series of Protrain Courses.=0D =0D NOW READY for Windows 95 on 3.5" disks=0D at =A349.50 each including post and packing and free updates for the firs= t two years =0D (=A358.16 inc VAT)=0D =0D M1 An Introduction to Scanning Electron Microscopy=0D This course covers - the CRT, magnification, image formation, spot size l= imitation, the electron gun, =0D lenses, deflection coils, the condenser system, focus & astigmatism, aber= rations, saturation methods, =0D spot size control, working distance, aperture alignment, image recording,= operating procedures, guide =0D lines to operation covering kV-spot size-magnification-working distance.=0D=
=0D M2 The SEM Specimen Preparation and Imaging=0D This course covers - the object of specimen supports, specimen mounts, fi= xing specimens in place, high =0D angle mounts, cross sections, reasons and materials for coating, sputter = coating, carbon coating, beam-=0D specimen reactions, reaction paths, reaction volumes, Monte Carlo simulat= ions, Everhart Thornley =0D detector, BSE detectors, image contrasts, image formation, specimen charg= e, =0D =0D M5 An Introduction to Transmission Electron Microscopy=0D This course covers - transmitted images, lens problems and resolution, el= ectron-specimen reactions, the =0D electron gun, the electron column, magnification and demagnification, def= lection coils, the condenser =0D system, beam coherence, condenser focus, astigmatism, image formation, fo= cus; diffraction, aberrations, =0D vacuum system, saturation, instrument alignment, eucentricity, photograph= ic calibration, the =0D photographic procedure, developing technique, an operating procedure.=0D =0D Available soon:-=0D M3 Operating Variables on the Scanning Electron Microscopy=0D M4 Advanced Scanning Electron Microscopy=0D M6 An Introduction to X-Ray Energy Analysis=0D M7 Intermediate Transmission Electron Microscopy=0D =0D These courses when combined with our popular "Protrain Portfolio" practic= al courses now make =0D it possible to "teach yourself" electron microscopy. TEM or SEM Portfoli= o contain all you need =0D (text book-instructions-specimens) to run a series of practical experimen= ts that will take the =0D student through intermediate and advanced instrument operating procedures= =2E =A3150 plus VAT =0D (=A3176.25 inc VAT)=0D =0D Check out our latest course and Multimedia offerings on the Web -=0D http://ourworld.compuserve.com/homepages/protrain=0D or e-mail us on protrain-at-compuserve.com=0D
With reference to EM training packages on CD.Contact Peter Goodhew. Mat Sci and Engineering at Liverpool Unin UK e-mail {goodhew-at-liv.ac.uk} who has some good stuff.
Patrick Echlin Multi-Imaging Centre University of Cambridge
On Tue, 19 Ma y 1998, Siegfried Jaecques wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear all, } } We are a university materials science department looking for } ways to make the EM training of our students (both graduate and } undergraduate) more efficient. We have a TEM and a couple of } SEM's, all from Philips (software controlled), and an old JEOL } SEM with EPMA (mostly manually controlled). } Currently the training of new users is handled by the chief } technician and is based on one-to-one 'hands-on' instruction. } However, with the number of users increasing rapidly, this is } becoming more and more of a burden on the chief technician. } Therefore, we are looking for ways to give the training to more } people at the same time and/or to give training without } occupying the instrument. } Our first idea was to look for instructional software packages, } preferably of the multimedia type (CD ROM based). } If you have experience with such software or know of information } sources about such software, your comments will be greatly } appreciated. Other solutions to our training problem may of } course also be suggested. Offers from commercial parties } (software vendors/EM vendors) are welcome. } We are also interested in instructional software on other } microscopic techniques (LM, AFM, AEM) and EDS and WDS, but this } interest is of a somewhat less urgent nature. } } Thank you, } } ------------------------------------------------------------ } dr. ir. Siegfried V.N. Jaecques } K.U. Leuven } Dept. Metallurgy and Materials Engineering (MTM) } de Croylaan 2 } B-3001 Leuven } BELGIUM } phone +32 16 32 1278 (direct) or +32 16 32 1260 (secretary) } fax +32 16 32 1991 } ------------------------------------------------------------ }
} One of the electrolytic polishing solutions used in the Tenupol Apparatus is } a mixture of 30% conc. nitric acid and 70% methanol.
Should such a mixture be stored, rather than made fresh each time? I do know that certain mixtures of ETHANOL and conc. nitric acid are rather unstable, decomposing after some minutes to give nitrogen dioxide among other things.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
John et al I made those pippettes for single cell injection as a junior tech well over 30 years ago. I guess they still use glass and that has not changed - much. Pull two Pasteur pipettes using about 120mm of capillary tubing. Hole may be 1-2mm diameter and wall thickness at least 2mm. Heat the tubing accross the narrow side of a fishtail burner as this will result in less heated area and a shorter tapered area. When sufficiently hot, hold the rod vertical and quickly pull to produce two pipettes. Heat the tip of the pipettes in a small flame to produce a small hook. To produce the final micro-pipette from these capillary pipette a simple commercial apparatus was used. A pipette was mounted vertically in a holder, a small weight was afixed to the hooked end. An exposed filament was moved very close to the the glass fibre capillary. The small weight could only drop by about 20 or so mm. The exposed filament would be brought to a certain heat setting and the weight would drop as the glass softened. I cannot remember how the end of the micro-pipette was finished off to produce a smooth tip. Maybe its just left rough but I suspect it is heated near that filament, while viewed under a low power scope. Hope that is some help. Its easy to make micro-pipettes, but to make them excellent is an artform. True of so much. Oh, since the question come from Australia, note we mounted the hook and weight down, because gravity sucks in the southern hemnisphere. Cheers Jim Darley
ProSciTech microscopy supplies and instruments email: service-at-proscitech.com.au fax: 61 7 47892313 online catalogue and information site: over 500 links www.proscitech.com.au catalogue, user notes, MSDS
} I dont know if this is the right place to look for responses, but; } Does anyone have a good reference that should be looked at to learn how o } pull your own micropipettes/needles and cell holders that you would use for } micromanipulation work. Specifically the removal of single cell contents } from plants? although I'm sure those used for mammalian cells would work. } Or is the experience that it's just better to buy them (Im in } Australia)? } } Any hints would be greatly appreciated. } } (im doing this for a colleague, so I dont know the specs of the equipment, } but I could find out. Yes we do have a microforge and other pieces of } equipment, we just haven't the experience to use them properly.) } } } }
Further to the point by Robert Olley, the ethanol/ nitric acid is unstable! It reacts after a short time rather explosivly. I used to employ this to clean coverslips for tissue culture; but times have changed. If you must use this method, place a 100ml beaker with about 5ml of nitric in a fumehood. Slowly add ethanol in 5ml glugs, pause when the beaker contains about 20ml total. When it turns yellow, draw the fumehood down. You can vary the type of alcohol used some more but the likely outcome is a quick trip to the hereafter. Stick with methanol/nitric for the original application please. Jim Darley
ProSciTech microscopy supplies and instruments email: service-at-proscitech.com.au fax: 61 7 47892313 online catalogue and information site: over 500 links www.proscitech.com.au catalogue, user notes, MSDS
} On Wed, 20 May 1998, Barry Searle wrote: } } } One of the electrolytic polishing solutions used in the Tenupol Apparatus is } } a mixture of 30% conc. nitric acid and 70% methanol. } } Should such a mixture be stored, rather than made fresh each time? I do } know that certain mixtures of ETHANOL and conc. nitric acid are rather } unstable, decomposing after some minutes to give nitrogen dioxide among } other things. } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: | } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } | University of Reading {University internal extension 7867 | } | Whiteknights Fax +44 (0) 118 9750203 | } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } | England URL: http://www.reading.ac.uk/~spsolley | } +------------------------------------------------------------------------+ } }
I am pleased to announce the website of the EUREM 12
12th EUROPEAN CONGRESS ON ELECTRON MICROSCOPY to be held in Brno, Czech Republic July 9 - 14, 2000
at http://www.eurem2000.isibrno.cz/
Please, forward this message to microscopists all over the world.
Best regards,
Petr Schauer +---------------------------------------------------------------------+ | Dr. Petr Schauer, Secretary of the tel.: (+420 5) 41514313 | | Czechoslovak Soc. for Electron Microscopy fax : (+420 5) 41514404 | | ACADEMY OF SCIENCES OF THE CZECH REPUBLIC (+420 5) 41514402 | | INSTITUTE OF SCIENTIFIC INSTRUMENTS e-mail: petr-at-isibrno.cz | | Kralovopolska 147, CZ-612 64 Brno csem-at-isibrno.cz | | Czech Republic www: http://www.isibrno.cz/csem/ | +---------------------------------------------------------------------+
just got around to fixing the vacuum situation on the 35. seems the problems were related to :
bad temperture sensor on DP badly broken down DP oil miscalibrated vacuum logic blackened and "plugged" DP stack
after some messy work (and an hour trying to figure out the jeol flange bolt puzzle!) it works well again.
Thanks to all who helped! This has been a good example (for me) of how we can collectively benefit using the net.
brian
btw: i just had a problem fixed on my cambridge S200 that has driven me crazy for awhile. seems the head amplifier on the 2-collector would breakdown after some period of usage....the image would deteriorate and become unusable. but as soon as you turned the scope off it dropped the supply to the amp and it would reset and be OK for some time again. thought it was the scintillator, lightpipe, apertures, sample, faraday cage bias, power supply, etc, etc, before stumbling onto the head amp...works wonderfully again!
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax) "Be well, do good work, and keep in touch"
Dear James, According to Szczesny, Walther and Mueller's paper in Current Eye Research (1996) the hexadecene is used to exclude gas bubbles (during High Pressure Freezing) and not as a cryoprotectant. They explain that hexadecene does not mix with water and therefore does not act as a cryoprotectant. It is suppose to be non-permeating or penetrating. In the literature, others do refer to it as a cryoprotectant (one example: Meindl et al, Protoplasma (1992) 170: 104-114). I have more references if you need them. beth
} A brief question: } Has anyone out there used hexadecene as a cryoprotectant? Could you advise } me on optimal times, and whether it is a permeating or non-permeating } agent? } } Cryogenically yours } } } James Wesley-Smith } Electron Microscope Unit } University of Natal } Durban, South Africa
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Dear Ritchie, I have made the Lion-S to Santovac-5 substitution on my Hitachi TEM (1982) and SEM (1986) successfully. It was recommended by the Hitachi Serviceman. I would watch the the 704 substitution, however, as 704 has a lower boiling point. I had problems trying to substitute 705 for 704 because the oil wouldn't boil. Check the poiling points in a vacuum catalogue such as Edwards or Precision. You wrote: } } So, I would like to know if anyone has successfully (and without } changing heater elements), made the following substitutions: } } 1 Octoil-S or Santovac instead of the recommended Lion-S in an old } (how old?) JEOL diff pump on a SEM, TEM, or EPMA? } } 2 Octoil-S or Santovac instead of the recommended Dow Corning 704 } in an Edwards 306 coater? } } thanks } } Ritchie } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
Dear Barry, This mixture of high concentration nitric acid in methanol is also used as liquid rocket fuel, so it is very flammable and dangerous. It can cause a runaway exothermic reaction, otherwise known as an explosion, as you mix it, so make it by slowly adding acid to methanol in a cooled bath and monitor the temperature to keep it cool ( {25 deg. C). I would personally never make such a large volume, if I could possibly help it. Store in a stoppered plastic bottle (not too tight) in a cool place and beware that these mixtures are prone to "let go" i.e. explode, for no reason at the back of the cupboard, so don't keep it too long ( {six months). When polishing, make sure you turn off the potential before removing the sample, to prevent any sparks between the sample and the bath. Does a lovely, fast job of polishing copper, though. You wrote: } } Fellow microscopists: } } In search of.................... } } One of the electrolytic polishing solutions used in the Tenupol Apparatus is } a mixture of 30% conc. nitric acid and 70% methanol. } } Excluding MSDS on each component - which contain some useful info see } Methanol "Safe Handling Procedures", what I am searching for is some current } practices for the proper storage of this mixture. } } Volume = 1000-1500 mL in a winchester } } and properly labelled .(*********************) } } How are other Users storing this chemical in the Laboratory? } } I am trying to obtain an MSDS of this mixture from Sturers - maybe it will } take a few days, if available at all. } } Does anyone have an MSDS on this mixture? } } } Thankyou for your help. } } } Barry } EM UNIT } UNSW Regards and good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
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I am not aware of a MSDS on nitric/methanol solutions.
This mixture is known to be unstable in some concentrations. If I remember correctly, the higher the nitric acid per cent the more unstable it becomes. It can be explosive.
We frequently use a mixture of 75% Methanol, 25% Nitric acid for chemical thinning. We do not store this reagent for more than one week. It is kept in a hood at all times, and never mixed with other chemicals.
Before disposing of the solution I would suggest you consult with your safety department regarding the best method of disposal.
If I can be of any further assistance, please feel free to contact me.
Carol
} } Fellow microscopists: } } In search of.................... } } One of the electrolytic polishing solutions used in the Tenupol Apparatus is } a mixture of 30% conc. nitric acid and 70% methanol. } } Excluding MSDS on each component - which contain some useful info see } Methanol "Safe Handling Procedures", what I am searching for is some current } practices for the proper storage of this mixture. } } Volume = 1000-1500 mL in a winchester } } and properly labelled .(*********************) } } How are other Users storing this chemical in the Laboratory? } } I am trying to obtain an MSDS of this mixture from Sturers - maybe it will } take a few days, if available at all. } } Does anyone have an MSDS on this mixture? } } } Thankyou for your help. } } } Barry } EM UNIT } UNSW
_______________________ Carol M. Garland, Member of the Professional Staff MC138-78 California Institute of Technology Pasadena, CA 91125
Could somebody help me resolve an issue that has developed in our lab recently? I realize this topic has been covered, but I can't seem to find a reference that addresses my specific question. It has been suggested that for purposes of ease that we store our EM samples in our glutaraldehyde fixative until we can process them. This suggestion truly unnerved me, because I have always been told that you should NEVER do such a thing for fear of 'over fixation'. Unfortunately when pressed, I could not provide an example of what overfixation means. All I have to go on is my gut feeling, the gut feelings of several of my colleagues, and conflicting information in a number of textbooks. Can anybody point me to a source that will definitively address this issue of overfixation in glutaraldehyde? Does overfixation really occur? If so, what are the specific artifacts caused by it? Or, is this all a myth? Your help will be greatly appreciated. Thanks, Laura
Laura M. Patrone, Ph.D. Wyeth-Ayerst Research Biomedical Imaging 641 Ridge Road Chazy, NY 12921 (518) 846-6318 e-mail: patronl-at-war.wyeth.com
George Lawton wrote: } } An investigator is having a problem with his procedure } on corrosive casting of rat kidney. After flushing the } kidney, it is infiltrated with methyl methacrylate. After } it is harden, the tissue is dissolved with conc. KOH. } He is getting crystals after dissolving. Has not had } this problem before. } Does anyone have an idea what the crystals are and } how to prevent them from forming. } Thanks in advance for any help. } } George Lawton
Dear George,
I bounced your question off Dr Fred Hossler, who is doing a corrosion casting session in Atlanta, and he offers the following:
I can only guess at the casting problem you passed on, but it sounds like a problem with salt precipitation either from the KOH or from his water source. Most casters now use 5% KOH (not concentrated KOH) to macerate tisseu becauseit was shown in a recent publication that (maybe Sims and Albrecht, but I forget the authors) this concentration actually macerates jsut as fast ashigher concentrations. I would also check if there are some mineral salts in the water that might be precipitatiing. In other words use distilled water for the maceration fluids. Let me know what happens. I am always interested in problems and solutions regarding casting.
I hope this helps. I can get you Dr. Hossler's e-mail address if you have any further questions.
JD Arnott Ladd Research
DISCLAIMER: Ladd Research sells corrosion casting materials
I recently read about a paper that was published concerning the determination of carbon coating thicknesses on EM samples without a thickness monitor. The paper stated that when utilizing a gold or polished brass sample, and certain parameters, the color change on the sample was indicative of a specific thickness in Angstroms. I was interested in finding this paper or any information related to this topic and would appreciate it if anyone who has knowledge of this topic would pass it on to me. Thanks.
Hi, Our laboratory is just starting to use Lowicryl for immunostudies. We need to buy a cryochamber. Question: How important is it to have a chamber with temperature control? Could we make do with a simple, less expensive chamber which uses dry ice without electronic temperature controls? What could we not do without accurate temp controls? Thanks, Hildy Crowley University of Denver
} Recently it came to my attention that he had processed a brain biopsy from } a suspected Creutzfeldt-Jakob disease patient. What really concerned me was } that : } 1) We had not been informed that the sample was in the lab } 2) The hospital technician did not appear to be aware of the high-risk } nature of the disease. } } My questions are: } } How do other labs that are have high risk samples going through them } monitor the risk of the samples coming into the lab? } } How do other labs handle and dispose of high risk biohazards? } Dear Allan & Richard, Working at a state health department gives us a leg up on such things. There are procedures in place for proper notification, handling & disposal. We know what samples are being brought in for examination, and since it is usually a physician who brings in any potentially danger- ous specimens, we are made aware of any potential hazards. Usually, the specimen has already been fixed, stained and embedded, which eliminates the hazard. There are containers all around our lab for the disposal of biohazards, and the safety office is responsible for their proper dispo- sal. Having the hospital technician give you a list of specimens and their condition (tissue, blocks, sections, etc.) before bringing them in would be an obvious step. I can think of rationalizations for not doing this, but maybe you can insist. If you have a safety office, by all means get them involved. Good luck. Yours, Bill Tivol
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
In a message dated 98-05-20 19:05:33 EDT, you write:
{ { Question: How important is it to have a chamber with temperature control? Could we make do with a simple, less expensive chamber which uses dry ice without electronic temperature controls? What could we not do without accurate temp controls? } }
Hildy,
Sure, you can use any kind of contraption as long as it will hold the desired temperature. We even used a styrofoam ice chest with dry ice in it. Get a large metal block that will fit in the ice chest, and drill holes in the metal that are big enough to hold small vials which would contain the specimens, alcohols, resin, etc. When the large metal block is pre-cooled it will help stabilize the temperature of the vials. You will have to do a little trial- and-error to find out how much dry ice to use and where to put it to hold the desired temperature.
If you have an old refrigerator the freezer compartment should be adjustable in the general range of temperatures for Lowicryl. Chest freezers also work well for this. Whatever you use, just make sure it's used exclusively for low-temp work with Lowicryl. The resin will permeate *everything*!
Agitation during dehydration and infiltration is fairly important. If you are working with an ice chest and metal block, just reach in and give a few manual "swirls" to each vial (or stir with a toothpick) about once every hour.
If you are working in a chest freezer, you can leave the door open and cut a sheet of building insulation the same size as the door. Presto, an instantly removable door. Then you can run a long axle at an angle from a slow-speed stirring motor through a hole in the insulation. On the end of the axle inside the chamber you can place a circular disk with clips on it for the specimen vials. In this way the stirring motor will stay outside the freezer at room temperature. It's probably not a good idea to put an entire specimen agitator in the freezer because water will condense and ice will form inside the agitator (electrical hazard!).
Consistent, stable low temperature and agitation are the keys. I've used all of the above and a few other configurations with good success.
Best wishes,
Bob ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Leica (Wild, Leitz, Bausch&Lomb, Cambridge, AO, Reichert-Jung) / Technical Instrument Company / American Volpi / Fostek / Stocker and Yale / AEI North America / OptiQuip / Dolan-Jenner / Osram / G.E. / Philips / Ushio / Boeckler Instruments / Heidenhain / Narishige / Colorado Video / Visual Environments of California, Inc. / Kinetic Systems / Pacific Precision Laboratories, Inc. / Pryor Scientific / Compumotor / Sutter Instrument Co. / Advanced Database Systems / Cohu / Javeline Electronics / Optronics / Diagnostic Instruments, Inc. / Dage MTI / Hitachi / Panasonic / Polaroid / Kodak / Mitsubishi / Sony
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Att. Sales / Export Department Re: Request for FIXATIVES
A "Request for supply" for FIXATIVES which, to the best of our knowledge are being offered by you, was placed with us by one of our clients.
We are a world wide sourcing firm and we are paid by our clients to find them suitable suppliers.
To you, our service is totally FREE OF CHARGE !!! The information we will get from you will not only be immediately sent to this particular client but also to other clients looking for same or similar items.
Since the whole operation is activated by our computers and unique software YOU HAVE TO REGISTER THE PRODUCTS ,MATERIALS, EQUIPMENT OR SERVICES you offer. Please REGISTER using our Internet interface at: http://www.thebol.com
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Att. Sales / Export Department Re: Request for FIXATIVES
A "Request for supply" for FIXATIVES which, to the best of our knowledge are being offered by you, was placed with us by one of our clients.
We are a world wide sourcing firm and we are paid by our clients to find them suitable suppliers.
To you, our service is totally FREE OF CHARGE !!! The information we will get from you will not only be immediately sent to this particular client but also to other clients looking for same or similar items.
Since the whole operation is activated by our computers and unique software YOU HAVE TO REGISTER THE PRODUCTS ,MATERIALS, EQUIPMENT OR SERVICES you offer. Please REGISTER using our Internet interface at: http://www.thebol.com
Once registered,the system will forward to you automatically the relevant to you requests.
At http://www.thebol.com you can also get more information about us and our FREE for SUPPLIERS SERVICE.
We are looking forward to serve you to the best of our ability.
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Att. Sales / Export Department Re: Request for FIXATIVES
A "Request for supply" for FIXATIVES which, to the best of our knowledge are being offered by you, was placed with us by one of our clients.
We are a world wide sourcing firm and we are paid by our clients to find them suitable suppliers.
To you, our service is totally FREE OF CHARGE !!! The information we will get from you will not only be immediately sent to this particular client but also to other clients looking for same or similar items.
Since the whole operation is activated by our computers and unique software YOU HAVE TO REGISTER THE PRODUCTS ,MATERIALS, EQUIPMENT OR SERVICES you offer. Please REGISTER using our Internet interface at: http://www.thebol.com
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Dear Microscopists, My apologies for the the message which just appeared on this list re CJD. This was an old message from Bill Tivol in response to my question about CJD risks for microscopists which I sent to this listserver about two years ago. I was sending my archived CJD messages to Joiner Cartwright (in response to his similar question here a couple of days ago) and didn't notice one of the messages had a 'Cc: microscopy-at-sparc5.microscopy.com' (copy to be sent to listserver). Oops.
Sorry also to Bill Tivol for broadcasting his old mail - but it his was a helpful message!
Incidentially, I haven't seen any responses to Joiner's message re CJD risks in microscopy labs this time around. His question was whether a consensus was reached in past listserver discussions on how to handle suspected CJD specimens - I don't think there was one. Our lab simply refuses to handle suspected cases but there must be labs out there that do do EM of CJD-infected tissue. It would be interesting to know what safety protocols they follow (for instance, do they do their ultramicrotomy in a fume hood?).
Best regards to all.
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
I don't know about the original paper, but Stephen Reed's book "Electron Microprobe Analysis" of 1975 has the following table for polished brass, quoting Kerrick, Eminhizer and Nakamura 1973: Amer. Mineral. 58 920
Thickness Colour (nm)
15 Orange 20 Indigo red 25 Blue 30 Bluish Green 35 Greenish Blue 40 Pale green 45 Silver gold
On Wed, 20 May 1998 16:23:28 -0400 "Wintonick, Steven" {WintonickS.bpd-at-ci.boston.ma.us} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I recently read about a paper that was published concerning the } determination of carbon coating thicknesses on EM samples without a } thickness monitor. The paper stated that when utilizing a gold or polished } brass sample, and certain parameters, the color change on the sample was } indicative of a specific thickness in Angstroms. I was interested in } finding this paper or any information related to this topic and would } appreciate it if anyone who has knowledge of this topic would pass it on to } me. Thanks. } } Steve
Institute of Physics 76 Portland Place, London W1N 4AA, UK. Tuesday 22 September 1998 IoP headquarters
Organizer: Dr U Bangert, Department of Physics, UMIST, Manchester M60 1QD, UK.
EMAG committee of the Institute of Physics
Abstract: Most technologically important materials are internally inhomogeneous, beit due to their 'natural' microcrystallinity (e.g. metal-alloys, complex oxides) or due to deliberate structuring (semiconductor nano-structures, nano-clustered materials). In fact, increasing complexity has been the key trend underpinning the development of electronic materials and devices for the past several decades. Hence there is an urgent requirement for the development of localized spectroscopies to work alongside localized structural determination. Depending on the scale of the material substructure spectroscopies down to the nm scale are desirable. Techniques based on modern scanning electron microscopes and scanning tunnelling microscopes are approaching this resolution, and it is hoped that this will contribute, for example ,towards an understanding of the electronic structure of grain boundaries, defects and nanostructures. At this meeting, experts in the field of spectroscopy based on the scanning (transmission) electron microscope and scanning tunneling microscope will report on new advances, with emphasis on the spatially resolved aspects.
Invited Speakers:
o R Brydson (Leeds) ' Chemistry on the nanoscale in the STEM'
o G Amaratunga (Liverpool) ' Electronic band-gap structure of amorphous diamond like and nano-structured carbon nanotubes determined by EELS in a STEM'
o C Norman (Toshiba, Cambridge), 'Cathodoluminescence of semiconductor films and device structures',
o B Hamilton (UMIST) 'Dopant and band gap imaging of semiconductors using STM'
o Struan Gray (Lund, Sweden) 'STM based photo-excitation microscopy'
o P Moriarty (Nottingham) 'Tunneling into single quantum dots'
o P Laitenberger (Oxford instruments) 'Low temperature scanning tunneling spectroscopy - probing solid state properties on the atomic scale'
Poster contributions are extremely welcome.
For further details please contact: Rebecca Chapple, IoP conference office (fax: 0171 470 4848, e-mail: rebecca.chapple-at-iop.org or Uschi Bangert, tel: 0161 200 3185, fax: 0161 200 3941, e-mail: uschi-at-fs2.phy.umist.ac.uk
_____________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
} I recently read about a paper that was published concerning the } determination of carbon coating thicknesses on EM samples without a } thickness monitor. The paper stated that when utilizing a gold or polished } brass sample, and certain parameters, the color change on the sample was } indicative of a specific thickness in Angstroms.
This sounds like D.M.Kerrick, L.B.Eminhizer & J.F.Villaume, "The role of carbon film thickness in electron microprobe analysis", American Mineralogist (1973), 58, 920-925
Using a piece of polished brass, they give a table of carbon coating thickness (in Angstroms) against interference colour:
150 - Orange 200 - Indigo red 250 - Blue 300 - Bluish green 350 - Greenish blue 400 - Pale Green 450 - Silver Gold
We have successfully used this technique in our laboratory for the last 20 years or so, and it's quite easy to judge the thickness to within about 25 Angstroms.
Norman
================================================= Dr. Norman Charnley Department of Earth Sciences University of Oxford Oxford OX1 3PR, UK.
Dear Sir: I am now doing some strain analysis in the herostructure ( one called LOCOS). At the moment, I only have result based on bulk materials. Do you know some good way to take into account the relaxation in this kind of semiconductor?
Hi, I'm just starting to use Quetol 561. I don't have any experience with this resin. I will be very appreciated with any imput that you can give me. Thanks, in advance.
FJ Iborra Sir William Dunn School of Pathology Oxford University Dr. FJ Iborra Sir William Dunn School of Pathology Oxford University South Parks Rd. Oxford OX1 3RE Tel. (+44/0) 1865 275527 Fax (+44/0) 1865 275501
We don't handle tissue of this type but I am surprised that glutaraldehyde does not sufficiently denature prions. Can anyone give me a key reference.
I had always assumed that normal tissue fixative (2.5-4% glutaraldehyde for 1+ hours) would render safe everything small and biological apart from some with impervious coatings (eg bacterial endospores, nematode eggs, some insects - tstse flies can swim in it for more than a day). Has anyone got a list of potential risks/awkward samples or a good source of information?
thanks
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ----------
I would like to know how medical EM labs handle specimens that potentially harbor Creutzfeldt-Jakob or other prion mediated diseases. We get very few calls to work up such specimens, but when we do, there is an awkward time deciding how to handle it, or whether to handle it at all. There is enough material in the literature on their resistance to fixatives and other agents that render bacterial and viral specimens safe to handle that people are quite reluctant to work with such specimens. I would like to hear from medical EM labs telling me whether or not you accept such specimens; and if you do, what special procedures do you use?
I understand that this might be a subject that has already been exhausted on the list server. However I have been off-line a while. If anyone remembers the consensus, I would appreciate a review. .........There HAS been a consensus on this server, hasn't there?
Joiner Cartwright, Jr., Ph.D. Director, Electron Microscopy Lab Department of Pathology
by piva.ucs.mun.ca (8.8.8/8.8.7) with ESMTP id JAA21053; Thu, 21 May 1998 09:57:50 -0230 (NDT) Received: from minnie.esd.mun.ca (minnie.esd.mun.ca [134.153.118.19]) by sparky2.esd.mun.ca (8.8.6/8.8.6) with SMTP id JAA17374; Thu, 21 May 1998 09:53:22 -0230 (NDT) Message-Id: {199805211223.JAA17374-at-sparky2.esd.mun.ca} X-Sender: rmason-at-sparky2.esd.mun.ca X-Mailer: QUALCOMM Windows Eudora Pro Version 4.0
Ritchie,
I substituted Santovac for the previous oil in a JEOL JXA 50A microprobe about 3 years ago. I did not change the heater element. The earlier oil (can't recall its name) tended to cause clogging of the diff. pump guts and to oxidise over a period of time: Santovac seems not to suffer from these deficiencies and I obtain a good vacuum.
Roger Mason
At 09:52 AM 20/05/98 +0000, Ritchie Sims wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
1630 - 1645 D.A. Jans Quantification of subcellular transport in single living cells using confocal microscopy: perforin-dependent nuclear entry of granzyme B in CTL targets precedes apoptosis
1645 - 1700 Y.E. Korchev Spontaneous and induced membrane dynamics: imaging with a scanning ion conducted microscope
1700 Finish
* Invited speaker
Registration Please contact the Royal Micorscopical Society for details of registration . Tel. 01865 248 768 Fax 01865 791 237, e-mail info-at-rms.org.uk or check our web-site at http://www.rms.org.uk
My full address is currently:
Dr. G. R. Coulton Molecular pathology Division of Biomedical Sciences Imperial College School of Medicine 8th Floor Lab Block Charing Cross Campus St. Dunstan's Road London W6 8RF
tel. 0181 846 7043 fax 0181 846 7099
e-mail g.coulton-at-cxwms.ac.uk
PLEASE NOTE THAT MY E-MAIL ADDRESS HAS CHANGED AND YOU SHOULD NOW MAIL ME AT
g.coulton-at-ic.ac.uk
HOWEVER, I WILL STILL RECEIVE MAIL TO THE CXWMS ADDRESS FOR A LIMITED PERIOD UP TO AUGUST 1998.
I am looking for a high magnification finite focus high NA water immersion objective preferably a Nikon PlanApo 40X to 63X. (Not a dipping lens) If anyone out there has converted to infinity optics. I will consider other manufacturers but would want to check it out first to make sure it will work.
Please contact me directly
Mike Esterman mikee-at-lilly.com 317-276-4247 7:30am - 4pm CDT
No viable information is in print about overfixation that specifically addresses the conditions of the tissue with glutaraldehyde. Several references allude to the problems, but fail to explain them. Articles in Ultrastructural Pathology and atlases of Pathology for Diagnostics have pointed out suspected artifacts.
For the record, based soley on my 30 years of working experience, when tissue is left in glutaraldehyde for extended periods, it becomes more difficult to section. The problem is identical to the difficulty with sectioning harder materials, i.e. skin, leading one to assume that 'overfixation' has created a hardened tissue condition. The presumed modification of artifactual appearance of the tissue in the microscope, when compared to tissue fixed for two hours or less, varies from none to moderate leaching of detail. Staining is less intense when compared to the routine times.
Our routine protocol calls for one hour fixation in 1% in PBS (350mOs, pH 7.2) then storage until processed in PBS (350mOs, pH 7.2). 90% of our samples are tissue culture. I would recommend 2% glut for minced animal or human tissue.
I need adress for manufactures for 120/220 roll film holder for optical microscopy - not polaroid !
best regards for all
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
Recently, I came across an article that discussed the handling of suspected Creutzfeldt-Jakob (sp?) disease specimens in a histology laboratory. It was an eye-opening article and mentioned a number of references that dealt with infectivity after fixation in formalin and paraffin embedding. Unfortunately, I have misplaced the article, but I do remember that it was printed in the newsletter edited by Don Grimes, called Microscopy Today, possibly from February or March 98. The web address for the newsletter is: www.microscopy-today.com.
Regards, Vicky Madden
Victoria J. Madden Dept. of Pathology and Laboratory Medicine University of North Carolina-Chapel Hill vmadden-at-med.unc.edu
With reference to cheap, easy to build cryochambers for Lowicryl, see pages 251-256 of my book "Low Temperature Microscopy and Analysis" Plenum Press New York 1992
Patrick Echlin
On Wed, 20 May 1998, HILDEGARD CROWLEY wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } Our laboratory is just starting to use Lowicryl for immunostudies. We } need to buy a cryochamber. Question: How important is it to have a } chamber with temperature control? Could we make do with a simple, less } expensive chamber which uses dry ice without electronic temperature } controls? } What could we not do without accurate temp controls? } Thanks, } Hildy Crowley } University of Denver } }
} I am looking for a high magnification finite focus high NA } water immersion objective preferably a Nikon PlanApo 40X } to 63X. (Not a dipping lens) If anyone out there has } converted to infinity optics. I will consider other } manufacturers but would want to check it out first to make } sure it will work.
I have a Zeiss Plan-Neofluar 63x/1.2 W in excellent condition.
You do not mention which Lowicryl you are planning to use, or during which stage (polymerization, infiltration, dehydration) you wish to control temperature.
We currently use a Reichert AFS freeze substitution device for controlling temperture for all these steps, but for Lowicryl K4M I think it is probably overkill. Dehydration and infiltration on ice (or in a -20C freezer after 50% EtOH) will probably work fine. For polymerization, we prefer to embed samples in flat molds for purposes of orientation. It is imperative not to expose the media to atmosphere, so we took an idea from Kent McDonald (Berkeley) and use a plastic pipette tip holder (it has a chamber on the bottom for holding dry ice, a perforated platform on which a beem flat embedding mold is placed, and a cover to keep the CO2 in and atmosphere out). This whole thing goes into a freezer and is exposed to UV for 24 hours. The dry ice sublimes over a 14 hour period or so, plenty of time for the media to polymerize enough so that oxygen is no longer inhibitory.
Good luck!
Doug
On Wed, 20 May 1998 13:43:55 -0600 (MDT) HILDEGARD CROWLEY {hcrowley-at-du.edu} wrote:
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- Send Email } to ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } Our laboratory is just starting to use Lowicryl for } immunostudies. We need to buy a cryochamber. Question: } How important is it to have a chamber with temperature } control? Could we make do with a simple, less expensive } chamber which uses dry ice without electronic } temperature controls? } What could we not do without accurate temp controls? Thanks, } Hildy Crowley University of Denver }
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
HELP! Does anyone have a technique for polychrome staining of TAAB embedded tissues? I am trying to find a connective tissue polychrome stain which I can use to stain semi-thins of cardiac valve before I do the good old TEM thing on the ultras. Any help will get a really nice mention in my thesis!
Hildy et al: In my opinion one should surely be able to scrounge up a cryochamber without spending money unless you simply want to do huge amounts ar have extra money. For example you could scrounge an old cryostat which is not in use and adapt it for lowicryl use. Or it may be possible to use a cold room depending on which formula of lowicryl you are using. It IS IMPORTANT to have some heat sink in your arrangment and this is where having a drilled aluminum block for your curing capsules can be very useful. We have in fact seen some incredibly high temperatures generated in the curing process when done with specific formulae and UV light distances with suspended block curing. Once you find a system that works well for you stick with it. Lowicryl is very noxious and toxic so bear that in mind when you construct your own set-up. Good luck in your construction. RM
A colleague would like to know if there is a way to convert his MRI files ( .FID ) that are in binary format to raw numbers for input into a spreadsheet. I don't know if this is the proper listserver to address MRI issues, but if it is not, maybe somebody could direct him to any known MRI listservers. He is using a Varian MRI with a SUN Solaris Sparc 20 workstation that is using Solaris 2.5.1 software. Please direct any respones to the Email address below:
jcsteve-at-ppco.com
Any information will of course be greatly appreciated.
Has anyone come across a methodology for determination of distribution of wax in particleboard, or know where I could search for such information? Apparently this can be done by using a wax soluble tracer and then using fluorescent microscopy, but I was wondering if there is any other method. Thanks for the help.
Jackie Terry ORTECH Corporation "Anything is possible when knowledge is shared."
"Handling Creutzfeld-Jakob Disease Tissues in the Histology Laboratory." by Michael Titford and Frank O. Bastian. MT, Feb/March '98 (#98-2), reprinted from the Journal of Histotechnology, Sept. 1983, vol. 12 #3.
Phil } } Recently, I came across an article that discussed the handling of suspected } Creutzfeldt-Jakob (sp?) disease specimens in a histology laboratory. It } was an eye-opening article and mentioned a number of references that dealt } with infectivity after fixation in formalin and paraffin embedding. } Unfortunately, I have misplaced the article, but I do remember that it was } printed in the newsletter edited by Don Grimes, called Microscopy Today, } possibly from February or March 98. The web address for the newsletter is: } www.microscopy-today.com. } } Regards, } Vicky Madden } } } } Victoria J. Madden } Dept. of Pathology and Laboratory Medicine } University of North Carolina-Chapel Hill } vmadden-at-med.unc.edu
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 5037 Station A Champaign, IL 61825-5037 USA oshel-at-shout.net or poshel-at-hotmail.com
The seriousness in the handling of CJ diseased tissues (survival of the agent after aldehyde treatment) is certainly evident after reading the following references. Anyone doubting the survival of such agents would do well to discuss this with Nobel Laureat, Prusiner, who discovered prions in the first place.
Take care with these tissues. One mistake could be fatal!!!
Prusiner, S.B.: The prion diseases. Sci Am 272:70-77, 1995
Telling, G.C., Scott, M., Hsiao, K.K., Foster, D., Yang, S.-L.l, Torchia, M., Sidle, K.C.L., Collinge, J., DeArmond, S.J., Prusiner, S.B.: Transmission of Creutzfeldt-Jakob disease from humans to transgenic mice expressing chimeric human-mouse prion protein. Proc. Natl. Acad. Sci. USA 91:9936-9940, 1994.
Cohen, F.E., Pan, K.-M., Huang, Z., Baldwin, M., Fletterick, R.J., Prusiner, S.B.: Structural clues to prion replication. Science 264:530-531, 1994.
Westaway, D., DeArmond, S.J., Cayetano-Canlas, J., Groth, D., Foster, D., Yang, S.-L., Torchia, M., Carlson, G.A., Prusiner, S.B.: Degeneration of skeletal muscle, peripheral nerves, and the central nervous system in transgenic mice overexpressing wild-type prion proteins. Cell 76:117-129, 1994.
Carlson, G.A., Ebeling, C., Yang, S.-L., Telling, G., Torchia, M., Groth, D., Westaway, D., DeArmond, S.J., Prusiner, S.B.: Eveidence for isolate specified allotypic interactions between the cellular and scrapie prion proteins in congenic and transgenic mice. Proc. Natl. Acad. Sci. USA 91:5690-5694, 1994.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I hate to admit that I forgot to add the catalyst to Spurr's epoxy prior to an attempted polymerization at 70C for 17 hours. The blocks have the consistancy of gumdrops now. If you have also done such a thing and salvaged your samples, would you please share with me what you did to make it right? I expect that several days at 80C should do the trick, but I have no experience.
Thank you,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
Jackie Terry wrote: ============================================= Has anyone come across a methodology for determination of distribution of wax in particleboard, or know where I could search for such information? Apparently this can be done by using a wax soluble tracer and then using fluorescent microscopy, but I was wondering if there is any other method. Thanks for the help. ================================================ We had to face this particular problem some number of years ago.
Actually we came up with a fairly simple kind of solution: We had our client make up two special waxes, one with dispersed carbon black particles and the other with a colloidal alumina (I think it was Ludox ® from DuPont). The particle board was then formulated with these two "special" waxes (which in those days were called low MW PE). By thin section TEM, the carbon black showed up better but it did not disperse as well. The Ludox dispersed better but it was not as easy to see (by TEM). But between the two approaches, it was possible to get some good insight as to where the low MW phase was going and to what degree the cross-section of the particle board was or was not asymetric.
I guess there was some argument as to what degree the carbon black or alumina added would have changed the rheology and therefore the viscosity of the wax. Valid arguments of course but we could not come up with any other bright ideas beyond these. The results for both methods seemed to be consistent and since there was an order of magnitude difference in size of the carbon black vs. Ludox, then what ever rheological change did occur must not have impacted on the final results.
There were some other issues such as the way the sample was embedded, etc., I would be happy to fill in some of the other details if you were interested .
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Regarding CJD and prion diseases, the handling of these tissues in the laboratory (and the mortuary!!) has excercised the minds of neuropathologists in Britain and Europe for some time. There is a "Consensus Report" from European neuropathologists regarding this subject (1), and the UK Advisory Committee for Dangerous Pathogens has also produced several reports (2). The Creutzfeld-Jakob Disease Surveillance Unit, in Edinburgh, also have protocols for handling these materials, including transportation and emergency response protocols.
(1) H Budka et al. Tissue handling in suspected Creutzeldt-Jakob disease (CJD) and other spongiform encephalopathies (prion diseases). Brain Pathology 5: 319-22 (1995).
(2) Advisory Committee on Dangerous Pathogens. Precautions for work with human and animal transmissible spongiform encephalopathies. HMSO, London (1994).
Richard Bonshek Dept of Pathological Sciences University of Manchester
I'm investigating microtwins in HgCdTe and CdTe as part of my M.Sc. project, but I'm having a little trouble indexing the twin spots in the diffraction patterns. I've used the method as shown in Hirsch, Howie and Whelan (1967), but I can't seem to get the method to work for some of the beam directions of the DP's. I was wondering whether anybody has an alternative approach or any suggestions to help me, since I am getting close to my wit's end with these things. The problem is that these materials are fcc and therefore twins should form on the {111} planes; analysis of these is supposed to be simple. Not so!
Any help/suggestions/accusations of stupidity etc will be appreciated. Thanks!
Doug, One of our techs did the very same thing. No amount of time in the heat will make it work. Our tech had to remove the specimen from the gummy Spurr's, make up fresh Spurr's and re-embed. Good luck!
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas ----------
I hate to admit that I forgot to add the catalyst to Spurr's epoxy prior to an attempted polymerization at 70C for 17 hours. The blocks have the consistancy of gumdrops now. If you have also done such a thing and salvaged your samples, would you please share with me what you did to make it right? I expect that several days at 80C should do the trick, but I have no experience.
Thank you,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
Any suggestions for maximizing the shelf-life of lithium-drifted silicon EDS-detectors during periods of time when keeping them filled with liquid nitrogen is not an option? For example, would keeping them cooler than room temperature help, and are there electronic ways to cool them at temperatures near LN2 (albeit still too warm to operate)?
Responding to the message of {B0000946530-at-gwarlmime.hmhs.com} from "Chism, Sharron" {SharronChism-at-hmhs.com} : } } Doug, } One of our techs did the very same thing. No amount of time in } the heat will make it work. Our tech had to remove the specimen from } the gummy Spurr's, make up fresh Spurr's and re-embed. Good luck! } } Sharron G. Chism HT (ASCP) } Electron Microscopy Lab } Harris Methodist Hospital } Fort Worth, Texas } ---------- } From: Doug Keene } To: Microscopy } Subject: Spurrs w/no catalyst } Date: Thursday, May 21, 1998 7:11PM
Sharon, Just what did your tech do ro remove the Spurr's resin, use a solvent, acetone? If you can't cure the resin without catalyst that resides in the sample, then must remove it somehow, then reinfiltrate with resin & catalyst. Any details you can give would be appreciated,
Thanks,
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
=========================================================== Jiri Spinka Faculty of Electrical Engineering and Computer Science Department of Electrotechnology Technical University of Brno EEEEEE TTTTTT Antoninska 1, B R N O EE TT Czech Republic EEEE TT Tel. 42-5-753741, Fax. 42-5-41211135 EE TT e-mail: spinka-at-uete.fee.vutbr.cz (Internet) EEEEEE(hi) TT ===========================================================
I've encountered a small 3-4 micron particle contaminate of metallic titanium in one of our processes. The source is unknown. TEM/EDX analysis of a cross section of this particle reveals a significant level of argon associated with the titanium and no oxygen. No, it's not a Ti escape peak, it's real. Can titanium contain sufficient dissolved Ar to register a significant peak? If this is true does it say anything about the source of this titanium or it's history? Thanks, Russ Russ_Gillmeister-at-wb.xerox.com
=========================================================== Jiri Spinka Faculty of Electrical Engineering and Computer Science Department of Electrotechnology Technical University of Brno EEEEEE TTTTTT Antoninska 1, B R N O EE TT Czech Republic EEEE TT Tel. 42-5-753741, Fax. 42-5-41211135 EE TT e-mail: spinka-at-uete.fee.vutbr.cz (Internet) EEEEEE(hi) TT ===========================================================
Modern Si(Li)s can be warmed without damage as long as they are not under bias. Should last for years at room temperature. The biggest concern is the cold finger may have condensed gasses on it. When the gasses evaporate upon warming the pressure may rise in the vacuum enough to blow the window. This would only happen if there has been a lot of gas, since the pressure would have to rise above atmospheric pressure to blow the window.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
Russ, Do you have a Ti sublimation pump or ion pump in the process? If so, the particle could have contained both elements as debris from the pump. However, I would have expected to detect oxygen, nitrogen and other gases as well.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
Russ writes ... } } I've encountered a small 3-4 micron particle contaminate of metallic } titanium in one of our processes. The source is unknown. TEM/EDX } analysis of a cross section of this particle reveals a significant } level of argon associated with the titanium and no oxygen. No, it's } not a Ti escape peak, it's real. Can titanium contain sufficient } dissolved Ar to register a significant peak? ...
I identified a similar particle in my microprobe. As it turned out the Ar was due to a leak in my flow detector and the ionization vacuum guage was gettering the argon inside its sensor ... creating metallic flakes with imbedded Ar.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
I know there have been MANY threads lately on digital cameras, which have been very informative. But there is such a boom going on out there! Does anyone have a good handle on the differences between the "megapixel" models which are marketed for consumer/professional photography markets vs. the scientific models discussed on this list??
Specifically, a colleague recently challenged me as to why I would want to "waste" $6000 on a Minolta RD-175 when I could get even better resolution from some of the Olympus, Kodak, Konica, Canon, etc etc. products out there for $1000 or less. After some web site searching, it seems like there might be an issue of true color representation (1CCD vs. 3CCD's) and perhaps lens mounts, auto-white balance w/o manual override, compression, or other things. But I'm not sure per particular model. Anyone want to get into some of these details??
NOTE: my needs are for Both hand-held close up work and possibly microscope-mount, to be used in subsequent image analysis of colors & densities as well as morphology. Low-light sensitivity not essential.
Some of the suggested models were:
High End: Minolta RD-175 Kodak DCS 420, 410, and 460 [anyone know a good vendor in Minnesota?] Polaroid DMC2000
Lower End or Unknown: Sony DKCST5 [again, any vendors in Twin Cities area?] Kodak DC120 or MDS120 Konica Q-M100 Olympus D-600L Canon EOS DCS 1 Pixera Professional [I have this] many others I haven't looked up yet
I hate to beat a dead horse, but this is more like cutting up a starfish. The issues just seem to multiply! Any input appreciated, Karen
-- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
Braun wrote: Dear colleagues , I am interested in some advices to prepare tungsten tips suitable for micromanipulation techniques. Who can help ?
Braun, a description to produce tungsten tips for micromanipulation tools can be found on pages 226 and 227 in volume I of the Particle Atlas (authors McCrone and Delly). If you do not have this book, send me your FAX number and I can send you a copy of the procedure.
Jackie Terry, ORTECH Corporation "Anything is possible when know-how is shared."
I am trying to make the stage of my Nicolet FTIR microscope positionable to within 10=B5m, and reproducible at this scale. The IR microscope is essentially built around an Olymbpus BH optical microscope platform, and uses Olympus' mechanical X-Y stage and slide holder ((U-SVRD) which has 1mm (0.1 mm) vernier calipers on X and Y axes that are locatable to only 100 =B5m. I would like to improve on this to 10=B5m or better. I do not really need (and cannot afford) an expensive stage with stepper motors and controllers, that typically give 1 =B5m accuracy. Does anyone have an idea on how to obtain a mechanical stage that is movable in 10=B5m or better increments? It can be manual or motorized, commercial or homemade, so long as it can be repositioned without actually having that location in view. I need only 20 mm of movement in either direction. Under $2,000 for purchase or modification would be preferable.
Thanks. Peter Michael
Please address any comments to me at pjm-at-utulsa.edu
Peter J. Michael Dept. of Geosciences 600 S. College Ave. Tulsa, OK 74104
Sorry, I left out the particulars! What she DID was to just re-embed in fresh Spurr's. There was SOME infiltration of the new Spurr's into the specimen, but not as good as it should have been. What SHOULD have happened (in my mind, at least) was to take the specimen out of the gummy Spurr's and back-track through a couple of changes of 100% ETOH, then a 1:1 Spurr's/100% ETOH step and then into a couple of changes of the new Spurr's before the final embedding step. I was on vacation at the time, so I have no way of knowing if that would have worked for sure ... but it sounds like it should.
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Hospital Fort Worth, Texas
Responding to the message of {B0000946530-at-gwarlmime.hmhs.com} from "Chism, Sharron" {SharronChism-at-hmhs.com} : } } Doug, } One of our techs did the very same thing. No amount of time in } the heat will make it work. Our tech had to remove the specimen from } the gummy Spurr's, make up fresh Spurr's and re-embed. Good luck! } } Sharron G. Chism HT (ASCP) } Electron Microscopy Lab } Harris Methodist Hospital } Fort Worth, Texas } ---------- } From: Doug Keene } To: Microscopy } Subject: Spurrs w/no catalyst } Date: Thursday, May 21, 1998 7:11PM
Sharon, Just what did your tech do ro remove the Spurr's resin, use a solvent, acetone? If you can't cure the resin without catalyst that resides in the sample, then must remove it somehow, then reinfiltrate with resin & catalyst. Any details you can give would be appreciated,
Thanks,
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
You might try the Desktop Microscopist program for the Macintosh. Among other things, it calculates single-crystal electron diffraction spot patterns for any crystal, and can use explicitly entered orientation relationships to calculate patterns from oriented crystallites.
You can check it out at http://www.easystreet.com/~lacuna/
I have no commercial interest in this program, but am a long-time user and sometime beta tester.
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA USA tel: 509 335-4860 email: thomas-at-mme.wsu.edu or (currently) Larry.Thomas-at-pnl.gov
---------- From: Alistair Douglas Reply To: phbaad-at-upe.ac.za Sent: Friday, May 22, 1998 9:16 AM To: Microscopy-at-Sparc5.Microscopy.Com Subject: Twin spots in TEM DP's
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Hello all
I'm investigating microtwins in HgCdTe and CdTe as part of my M.Sc. project, but I'm having a little trouble indexing the twin spots in the diffraction patterns. I've used the method as shown in Hirsch, Howie and Whelan (1967), but I can't seem to get the method to work for some of the beam directions of the DP's. I was wondering whether anybody has an alternative approach or any suggestions to help me, since I am getting close to my wit's end with these things. The problem is that these materials are fcc and therefore twins should form on the {111} planes; analysis of these is supposed to be simple. Not so!
Any help/suggestions/accusations of stupidity etc will be appreciated. Thanks!
You didn't mention what your process was, but it's not uncommon to find a significant quantity of included process gas in thin film depositions. I first observed this in my graduate work with rf sputtered depositions of Germanium. Sputtering Germanium with Argon, under certain conditions, can result in films with greater than 5% argon (at %). Any similar process including plasma processes, sputter depositions, ion bombardment, anomalous arcing, etc. may result in coatings or particulates with included process gas.
Phil Swab Advanced Coatings Division/ART Buffalo, NY
} ---------- } From: Gillmeister,Russ[SMTP:Russ_Gillmeister-at-wb.xerox.com] } Sent: Friday, May 22, 1998 10:27 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Titanium } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } I've encountered a small 3-4 micron particle contaminate of metallic } titanium in one of our processes. The source is unknown. TEM/EDX } analysis of a cross section of this particle reveals a significant } level of argon associated with the titanium and no oxygen. No, it's } not a Ti escape peak, it's real. Can titanium contain sufficient } dissolved Ar to register a significant peak? If this is true does it } } say anything about the source of this titanium or it's history? } Thanks, Russ Russ_Gillmeister-at-wb.xerox.com }
I once found argon in a metal substrate which had been subjected to some sort of vacuum depostion process. I don't remember any of the details (age or feeble mindedness?) except that I tested a lot of samples and did a lot of soul-searching before I let anyone know that's what I found. As it turns out, argon was used in the process and I assume that some argon was trapped in the surface layer.
Your particle may be due to some such deposition process.
Thanks Robert A. Carlton Robert.Carlton-at-RP-Rorer.com Tel. 610-454-3949 Fax 610-454-5990
As sometimes happens, some people can't get money to attend courses at the last minute while others don't hear about them until it is too late to apply.
Now may be your chance!
Three late cancellations at the UBC Live-Cell course mean three opportunities for those of you who have ten days free this June.
We expect to have an international faculty of 15, and on-site equipment that includes 2-photon systems from BioRad, Zeiss, and Olympus (and maybe Nikon and Leica). There will also be single-photon confocals from Bio-Rad (2), Olympus, Noran, Nikon (2), deconvolution set-ups from API, AutoQuant and Vaytek as well as laser tweezers, micromanipulation and sundry other delights.
These systems are not just to look at but to use for over 45 hours or organized 2D and 3D live-cell laboratory sessions, and 20 hours of evening sessions for group live-cell projects.
Although the eight "groups-of-3" have already been set up and have chosen their "individual projects," we are willing to accept 3-2 more students as long as their backgrounds will fit into any of these groups.
which has the rest of the story, including the program.
Then fill out the Registration Form from the site to tell us about you, Fax it to me and we will see if we can fit you in.
Hope that you can join us in Vancouver this June 17-28.
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
I wrote: I'm going to stick my neck out here, but I was able to produce this in a sputtering system. Surprisingly, burning a hole(melting) in the top of blisters caused the argon signal to appear and disappear. I believe we were playing with an experimental RIE(Reactive Ion Etching) system. I know what some of you are thinking(me too),even though I observed this and repeated this several different ways I somehow still have trouble believing there wasn't another explanation. -Anyone else? Jeff Day -"JD" Mesquite, Texas
note: Well, it seems I must explain how deposition occurred in RIE, before someone(else) thinks that I might not know what I'm talking about. Our experiments appeared to suggest that under certain circumstances(targets of multiple elemental components and some poorly chosen vacuum components) both etching and re-deposition were taking place in RIE especially in an argon environment and generally at higher powers. I don't think this is really new information to anyone with RIE experience.
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
I have attempted and failed twice to store these detectors at room temperature. Additional to the problem indicated by Mark Lund, the contaminants collected by the absorbing material is released at room temperature and this contaminates the Si(Li) wafer, making it later unusable. I had been successful re-pumping, including giving the heat treatment (which should remove contaminants) a "soft detector", but storage for some months proved unsuccessful. This was with detectors produced over 15 years ago. Has there been a change in detector technology that would affect storage? Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** - } Modern Si(Li)s can be warmed without damage as long } as they are not under bias. Should last for years } at room temperature. The biggest concern is the } cold finger may have condensed gasses on it. When } the gasses evaporate upon warming the pressure } may rise in the vacuum enough to blow the window. } This would only happen if there has been a lot } of gas, since the pressure would have to rise } above atmospheric pressure to blow the window. } } best regards } mark } } Mark W. Lund, PhD } Director } } Soft X-ray Web page http://www.moxtek.com { { } MOXTEK, Inc. } 452 West 1260 North } Orem UT 84057 801-225-0930 FAX 801-221-1121 } lundm-at-xray.byu.edu } } "The state is good at simple tasks, like killing people and seizing their } wealth. It has far more trouble reaching inside individuals and making them } good." Doug Bandow } }
I was looking for a listserve with discussions on condensed matter. Do not respond to the listserver, respond to the mail cited below. Thank you in advance.
Gabriel A. Rosa e-mail : micros-at-biolo.bg.fcen.uba.ar FAX : (0054-1) 788-6770
Dear Karen, There are good reviews of digital cameras in several magazines: Try Popular Photography (Dec. 1997), PEI - Photo Electronic Imaging (May 1998) www.peimag.com, and PC Magazine (February 1998).
We are thinking of getting the Olympus D-600L. Please note that PC Mag reviews the D-500L not the 600.
Once we narrowed it down to the D-600L I got price quotes from National Graphics Supply 800-223-7130, B&H 800-947-5516, and Wolf Camera (the local Wolf Camera would only quote me list price- so poop on them).
Hope this helps, beth
} To the digital die-hards out there: } } I know there have been MANY threads lately on digital cameras, which } have been very informative. But there is such a boom going on out } there! Does anyone have a good handle on the differences between the } "megapixel" models which are marketed for consumer/professional } photography markets vs. the scientific models discussed on this list?? } } Specifically, a colleague recently challenged me as to why I would want } to "waste" $6000 on a Minolta RD-175 when I could get even better } resolution from some of the Olympus, Kodak, Konica, Canon, etc etc. } products out there for $1000 or less. After some web site searching, it } seems like there might be an issue of true color representation (1CCD } vs. 3CCD's) and perhaps lens mounts, auto-white balance w/o manual } override, compression, or other things. But I'm not sure per particular } model. Anyone want to get into some of these details?? } } NOTE: my needs are for Both hand-held close up work and possibly } microscope-mount, to be used in subsequent image analysis of colors & } densities as well as morphology. Low-light sensitivity not essential. } } Some of the suggested models were: } } High End: } Minolta RD-175 } Kodak DCS 420, 410, and 460 [anyone know a good vendor in Minnesota?] } Polaroid DMC2000 } } Lower End or Unknown: } Sony DKCST5 [again, any vendors in Twin Cities area?] } Kodak DC120 or MDS120 } Konica Q-M100 } Olympus D-600L } Canon EOS DCS 1 } Pixera Professional [I have this] } many others I haven't looked up yet } } I hate to beat a dead horse, but this is more like cutting up a } starfish. The issues just seem to multiply! } Any input appreciated, } Karen } } -- } Karen Zaruba, kszaruba-at-mmm.com } BioMaterials Technology Center } 3M Center Bldg. 270-1S-01 } St. Paul, MN 55144 } } *The opinions above are my own, not necessarily my employer's*
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I am interested in obtaining floorplans and/or suggestions for an efficient, functional forensic microanalysis/trace evidence unit. I've gotten the EM lab design info that was mailed lately amongst the group. Thankyou.
Alice Ammen Montana Forensic Science Division 554 West Broadway, 6th floor Missoula, MT 59802
Does anyone know of and have any positive experience with any good sources of 3-D reconstructive software for serial section data? Shareware or Commercial? If you are a vendor I suppose this list server would prefer that you respond to my Email directly. I'm also interested in some secondary manipulative software that can rotate and change(distort-attenuate or amplify) axis in three(3) or five(5), ie. two( 2) or more rotational axis. Any other suggestions? Jeff Day/ "JD" Mesquite, Texas Email: wa5ekh-at-juno.com
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
Hi all, We have looked at this problem of mothballing detectors for clients = going on long leave or over the Christmas period, and in some cases for = indefinite periods. Realising that you get this outgassing of the molecular sieve as the = detector warms up, we made up a T piece pumping port adapter for the = detectors.
The secret to storing the detectors is the pumping of the detector when = you allow it to cool down. This results in you pumping off all the = contaminants in the dewar before it can settle on the crystal and FET. If you look on the side of the detector, you will see a red or blue = cap. Open that and you will see the plug that needs to be removed to = pump down the detector. The pumping port adapter can be made up by your local engineering = workshop.=20 One warning, we suggest you start the pumping down the minute you throw = off the LN2. On two occasions we have had the vacuum plug blow out about = 30 minutes after pouring off the LN2 . So in some detectors it seems = that as they warm up, they build up a positive pressure. You would = expect the Be window to blow first, but not so. Exactly why we are not = sure.
The pumping system must be able to attain at least 10-5 torr so either = diff or turbo systems will do. If there is a LN2 trap in the system, = that must be used and is ideal. If the SEM is also not going to be used for a week or two, you could = make a blanking port with a vacuum port for the SEM chamber and this = would allow you to use the SEM as the pumping system. Not ideal but it = is a way round it. Connect the vacuum system to the detector and once the system has = reached 10-1 torr, remove the vacuum plug on the detector. We generally = open the plug with only the rotary pump and not under diff pump = pressures. This is because some detectors have a really poor vacuum and = you could damage the diff pump if you opened the detector on to it. = Leave it to pump for a minimum of 24 hours. Once the detector is up to room temperature, you can fill the dewar with = boiling water. Watch the vacuum gauges at this point. In cases where the = dewar is really contaminated you will see a drop off in vacuum as soon = as the warm water is poured into the dewar. You can repeat this process = until the vacuum does not change when you add hot water. Once the water in the dewar has returned to room temperature, you can = now carefully close the pumping port to the detector and store the = detector. This is a quick description of what is involved in the process of = storing detectors. I hope it make sense. Failing this you can simply ask your supplier to do this for you and PAY = THEM FOR IT. Good luck
Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
-----Original Message-----
I have attempted and failed twice to store these detectors at room temperature. Additional to the problem indicated by Mark Lund, the contaminants collected by the absorbing material is released at room temperature and this contaminates the Si(Li) wafer, making it later unusable. I had been successful re-pumping, including giving the heat treatment = (which should remove contaminants) a "soft detector", but storage for some = months proved unsuccessful. This was with detectors produced over 15 years ago. Has there been a change in detector technology that would affect = storage? Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** - } Modern Si(Li)s can be warmed without damage as long } as they are not under bias. Should last for years } at room temperature. The biggest concern is the } cold finger may have condensed gasses on it. When } the gasses evaporate upon warming the pressure } may rise in the vacuum enough to blow the window. } This would only happen if there has been a lot } of gas, since the pressure would have to rise } above atmospheric pressure to blow the window. } } best regards } mark } } Mark W. Lund, PhD } Director } } Soft X-ray Web page http://www.moxtek.com { { } MOXTEK, Inc. } 452 West 1260 North } Orem UT 84057 801-225-0930 FAX 801-221-1121 } lundm-at-xray.byu.edu } } "The state is good at simple tasks, like killing people and seizing = their } wealth. It has far more trouble reaching inside individuals and making them } good." Doug Bandow } }
Question: Looking for a quick, proven recipe for fixation (for TEM) of cells in a suspension culture. They're a line of chronic myelogenous leukemia in IMDM media w/ 10% FBS. Problem is, I'd like to avoid any spinning down/resuspension techniques as I am attempted to capture the cells after they have been induced into apoptosis. There's a lot of interesting morph. changes in the cell membrane (blebbing of apoptotic bodies) that are very fragile and I'd like to preserve them if possible. Right now, I've been embedding the suspension in agar and processing the agar blocks. I just haven't had any decent results yet. Any experts out there?
charles j day wrote: } } Does anyone know of and have any positive experience with any } good sources of 3-D reconstructive software for serial section data? } Shareware or Commercial? If you are a vendor I suppose this list server } would prefer that you respond to my Email directly. I'm also interested } in some secondary manipulative software that can rotate and } change(distort-attenuate or amplify) axis in three(3) or five(5), ie. } two( 2) or more rotational axis. Any other suggestions? } Jeff Day/ "JD" } Mesquite, Texas } Email: wa5ekh-at-juno.com
Hi Jeff,
I don't know of any specific software for serial section reconstruction, but You could certainly do it with a powerful image processing package. AVS is a commercial one and Khoros is probably the most powerful free one (at least the current version 2.2 is still free). You can stack sections, align them, distort them, change scale and rotate them, but You have to make your own algorithms by piecing together processing tools in a workspace. (This is similar to AVS) Khoros handles up to 5-dimensional data so you can make colour-movies of 3D-models if you need to. Look at: www.khoral.com for more information. There's also a very active mailing list and news group to help you. -- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
---------- From: Alistair Douglas Reply To: phbaad-at-upe.ac.za Sent: Friday, May 22, 1998 9:16 AM To: Microscopy-at-Sparc5.Microscopy.Com Subject: Twin spots in TEM DP's
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Hello all
I'm investigating microtwins in HgCdTe and CdTe as part of my M.Sc. project, but I'm having a little trouble indexing the twin spots in the diffraction patterns. I've used the method as shown in Hirsch, Howie and Whelan (1967), but I can't seem to get the method to work for some of the beam directions of the DP's. I was wondering whether anybody has an alternative approach or any suggestions to help me, since I am getting close to my wit's end with these things. The problem is that these materials are fcc and therefore twins should form on the {111} planes; analysis of these is supposed to be simple. Not so!
Any help/suggestions/accusations of stupidity etc will be appreciated. Thanks!
Kind regards Alistair Douglas
I'd like to make the following suggestions that might help:
Besides the book of Hirsch et al. published in 1967, please have a look at the followings: - the same book, but in its new version published in 1977 at Robert E.Krieger Publ.Co., pp.143-148 & 532-534 & 545-546; - the book of Gareth Thomas and Michael J.Goringe "TEM of Materials", published at Wiley Interscience (I don't know the year of issue), pp.94-100; - C.Boulesteix, J.van Landuyt and S.Amelinckx, "Identification of Rotation and Reflection Twin by Diffraction and Contrast Experiments in the Electron Microscope", published in Physica Status Solidi, A33, 595 (1976);
and I can give you a much longer bibliographical list on twinning if you are interested in.
As concerns the software mentioned by Larry Thomas in his public message, I have no doubt it is useful, but you have to buy it !
I put before you a proposition: I have recently developed a computer code for simulation of the ED patterns of any complexity (including the effect of rod-spiking - described by Hirsch et al., in their book above mentioned by yourself) and in case you agree to send me all the geometrical data concerning the spots distribution in your ED pattern and the crystallographic data of your crystalline compounds HgCdTe and CdTe I can try to give a solution to your problem. Of course, I might send you my soft - in the executable form (it runs on IBM-PC under MS-DOS), but it is a little too much complicated to explain you how to use it. My computer program can simulate by calculation and display graphically a complex ED pattern which can include also twinning effects, these complex patterns being generated by superposition of simpler ones, according to any twinning model you can imagine (for any arbitrary crystal orientation, beam direction etc.). I am quite sure that the software recommended by Larry Thomas works in the same way as mine.
Corneliu Sarbu National Institute for Materials Physics POBox MG-7 R-76900 Bucharest-Magurele Romania Fax: +40-1-423.1700
A couple of months back a message you posted to the listserver said that you had an assortment of addresses and quotes pertaining to EDS upgrades. I would appreciate it very much if you could send that information to me.
We are looking for an unused quantity of a Chemicon antibody (anti-Tenascin (Hu - polyclonal) Cat. No. AB1906) which has been discontinued. We have been getting good localisations with it in IHC experiments using rat tissues. Chemicon have informed us they do not have a substitute for it, so we are basically back to square one. If anyone has some left in a fridge somewhere we would be happy to pay for it plus express freight costs. Also if anyone has experience with this antibody and have since found a replacement or any other information, please let us know.
Also does anyone have any experience with SPARC (osteonectin)?
This is a question for the microscopists who deal with paper samples:
Does someone have experience of using Monte Carlo simulations for determining the penetration depth of the beam into such a complex system like paper is? Do you see any advantage in using it? I'm interested in backscatter imaging of coated paper surface and I'd like to know what depth I'm actually looking at.
Thanks a lot.
Tiina Hallamaa, M.Sc (Eng) The Finnish Pulp and Paper Research Institute Paper Science Center email: Tiina.Hallamaa-at-kcl.fi
I have had good results using T3D (formerly called Slicer) from Fortner Research, LLC. (100 Carpenter Drive, Sterling, VA 20164, (703)478-0181, www.fortner.com) to visualize serial sections.
What I like about T3D is that it will automatically read in a sequentially numbered set of TIFF files (you open the first one and T3D will find the rest). You can independently adjust the scale on the three axes to adjust for aspect ratios and slice thicknesses. T3D can also create AVI animations of the visualization.
Everett Ramer U.S. Department of Energy P.O. Box 10940 Pittsburgh, PA 16236-0940
A summary of 3-D imageing and reconstruction programs was posted on the list about a year ago. Here it is again. I hope the original author of this doesn't mind but I have found this to be very useful information
First I want to thank everybody who responded to my question on suitable software to reconstruct the 3D-structure of organs (based on serial sections). I think it's a good idea to summarize all the tips I have got the last week. The list below is far from beeing complete (sorry to everybody who thinks that I have forgot something important).
NIH-IMAGE This popular freeware programme for image analyses is originally written for the Mac, but now is also available as Win95 program. Download: http://www.zippy.nimh.nih.gov/ (Mac)or http://www.scioncorp.com/ (Win95) Many informations e.g. online manual, macros, example-files and additional download possibilities can be found at: http://rsb.info.nih.gov/nih-image/ As an example animated reconstructions of plant cells (based on semithin sections) can be found on the homepage of Gary Chinga: http://www.nvg.unit.no/~gary Informations about the NIH-Image mailing list can be found at http://www.soils.agri.umn.edu/infoserv/lists/nih-image/
NEUROLUCIDA Informations about Neurolucida are available at http://www.microbrightfield.com/microb Although Neurolucida is primarily designed for the needs of neurobiologists, it has all the facilities which I need for the planed reconstruction based on serial tissue sections (Data entry by camera or digitizing tablet, alignment, tracing of areas of interest, 3D-modelling and volume measuring).
VOXBLAST Voxblast is available for Unix, Mac and Windows-Sytems. Informations about this software, e.g. instructive demo-versions and a quick-reference-quide can be found at http:/www.vaytek.com/ In order to transfer and prepare (enhancement and alignment of the slices) the images for the use with Voxblast, suitable image analyses software, e.g. NIH-image or ImageProPlus from Mediacybernetics (http://www.mediacy.com/) is needed.
T3D (formerly SLICER) Included in the data-management-package NOESYS of Fortner research. Product informations can be found at http://www.fortner.com/
SLICER DICER Informations about this volumetric data visualization software of Visalogic (available for MAC, Win95 and WinNT) could be found at http://www.visualogic.com/
MONTAGE This 3D reconstruction package is a UNIX-programm, but works also on a PC using the free Unix-system LINUX. It's available by anonymous FTP at ftp://retina.anatomy.upenn.edu:/pub/mont.linux.tgz Data entry is simply by a Summagraphics Digitizing Tablet.
A list of links to the distributors of the above mentioned software and many other programs for image analyses can be found at: http://ddsdx.uthscsa.edu/dig/sites.html
This is all I get up to now from the internet and of course I haven't tested all the programs listed above for their suitability.
Sincerely,
Jens Buecking
--------------------------------------------------------------- Dr. Jens Buecking Tel.: +49-(0)421-218 3745 University of Bremen Fax.: +49-(0)421-218 4042 Department of Biology email: jbueck-at-biologie.uni-bremen.de Leobener Str. - NW2 D- 28359 Bremen Germany ---------------------------------------------------------------
Dr. Heike Buecking University of Bremen UFT Physiological Plant Anatomy Leobener Str. D 28359 Bremen Germany TEL: +49-421-218-2954 or TEL: +49-421-218-7283 FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de
Sorry, don't have the ab, but have you checked Linscott's Directory of Immunbological and Biological Reagents ISBN: 0-9604920-4-6, 4877 Grange Rd., Santa Rosa, CA 95404, 707 544-9555.
I highly recommend purchasing a copy of this book. It lists scads of antibodies and the companies that produce them.
I have no commercial interest in marketing this reference.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Greetings All! Does anyone have a procedure for the Von Kossa Stain that will work on Spurr's embedded tissue? We have a bone specimen that has not been put into decal solution, but has been processed and embedded in Spurr's. I have cut a "thick" section and have a slide ready to stain, but need a procedure for plastic embedded tissue.
Thanks in advance,
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Hospital Fort Worth, Texas
} Does anyone know of and have any positive experience with any } good sources of 3-D reconstructive software for serial section data? } Shareware or Commercial? If you are a vendor I suppose this list server } would prefer that you respond to my Email directly. I'm also interested } in some secondary manipulative software that can rotate and } change(distort-attenuate or amplify) axis in three(3) or five(5), ie. } two( 2) or more rotational axis. Any other suggestions? } Jeff Day/ "JD" } Mesquite, Texas } Email: wa5ekh-at-juno.com }
My name is Patrick Guerin. I work for VayTek where we produce and sell imaging software.
I'm responding to a question from Jeff Day posted on the list. I'm responding on the list because this question comes up from time to time, as membership to the list is dynamic, there are always new members joining. This informationmay be usefull for others.
} Does anyone know of and have any positive experience with any } good sources of 3-D reconstructive software for serial section data? } Shareware or Commercial? If you are a vendor I suppose this list server } would prefer that you respond to my Email directly. I'm also interested } in some secondary manipulative software that can rotate and } change(distort-attenuate or amplify) axis in three(3) or five(5), ie. } two( 2) or more rotational axis. Any other suggestions? } Jeff Day/ "JD" } Mesquite, Texas } Email: wa5ekh-at-juno.com
We have a 3D Volume Visualization and Measurement software that performs what you asked and more. There are several products that can take a stack of 2D images and create a volume that can be rotated.
Our product called VoxBlast has many unique features and functions that allow you to cut, fade, hide, rotate, translate, shade, extract, measure 2D and 3D, animate, and much more. We also have a module that can take the images from a scanner and automatically normalize the brightness, cute out the slides, center them and generate the appropriate volume file for VoxBlast. I would love to explain all these in detail, but fear that I would tax the patience of many members.
If you have an interest in such software, please contact us directly or visit our web site: WWW.VAYTEK.COM
Best regards
Patrick Guerin Customer Technical Support Engineer VayTek, Inc. 305 West Lowe Avenue, Suite 109 PO Box 732 Fairfield Iowa 52556-0732 Tel : 1-515-472-2227 ext. 103 Fax : 1-515-472-8131 WWW.VAYTEK.COM E-mail : pguerin-at-vaytek.com
Chris MacLean, Ph.D. VayTek, Inc. 305 West. Lowe, Suite 109 P.O. Box 732 Fairfield, Iowa, 52556-0732 Tel: 515-472-2227 ext.105 Fax: 515-472-8131 E-Mail: cmaclean-at-vaytek.com Web: www.vaytek.com -----Original Message-----
CENSUSCD+MAPS REVOLUTIONIZES DEMOGRAPHIC MAPPING
GeoLytics compresses 75 CD-ROMs of demographic data and boundaries onto ONE easy-to-use Windows disc with complete mapping software.
East Brunswick, NJ, March 31, 1998 - GeoLytics announces the release of CensusCD+Maps--a demographics and mapping software product combining 50 gigabytes of data with advanced thematic mapping capability.
CensusCD+Maps lets anyone create colorful thematic data maps, down to the neighborhood level of census block groups, with no mapping or GIS experience required. All of the data, boundaries, and software to create results are on the one disc. CensusCD+Maps eliminates the hassles of importing data and boundaries required by most mapping software.
The one Windows (95, NT, or 3.1x) CD-ROM of CensusCD+Maps provides:
- Estimates (1997) & Projections (2002) of Demographics (65 variables for each year) and Consumer Spending (32 product categories for each year) for 5 geographic levels (block group, tract, zip, county & state)
- 1990 Census (STF-3) demographics, over 3,400, for each of 375,000+ geographic units including block groups, tracts, and zip codes
- Integrated Thematic Mapping with complete boundaries (from Tiger 95) down to block group geographic level. A built-in map viewer creates and displays thematic maps of .DBF files generated by the program. The process is seamless for the end user. Maps are easily customized (themes, colors, ranges, scales, etc.) on the fly. A virtual variable calculator lets user perform mathematical functions (incl. Boolean) then automatically maps results. Viewer exports boundaries in ArcView & MapInfo, and maps as BMP. It thematically maps any imported .DBF file with an appropriate area key.
- County Time Series Statistics - 3,000+ variables in 26 categories for all 3,141 US Counties. Data is from US Govt. Agencies on topics such as Federal Spending (AFDC, SSI, Grants, Contracts, Payroll, etc..), Crime (FBI stats), Agriculture, Business (types, earnings, employment, payroll), Banking, Building Permits, Vital Statistics (births, deaths, etc.) and much more.
- Historical Census Counts (Total Population 1790 - 1990) by US County
- Geocoding of an address to its corresponding census block group to extract neighborhood information. Also allows for radius reporting around a specific address for any number of miles
GeoLytics first product, CensusCD 1.1, holds the complete results of the 1990 US Census long form (STF-3), originally issued on 67 CDs. GIS World noted in a March, 1997, review of CensusCD 1.1, "Performance of the demographic queries is outstanding. If all databases of this size running on CD-ROM performed this well, the information systems world would be a much better place." GeoLytics improved its compression technology to guarantee CensusCD+Maps lives up to the same performance standards.
GeoLytics sells CensusCD+Maps directly for $249.95 for a single user license; $750 for LAN/CD-Tower license. Please visit the web site at http://www.censuscd.com/cdmaps/censuscd_maps.htm for more detailed information.
GeoLytics can be reached at 800-577-6717 or by e-mail at info-at-censuscd.com.
Is there a fluorescent marker that will stain the cell membranes ( rater than the nuclei) ? Thank you, Lilith -------------------------------------------- Lilith Ohannessian-Barry NRC, IBS, Ottawa, Ont. K1A 0R6 Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
Hi, Is anyone using an air purifying system in their lab or darkroom? A local business gave me a Bora Air Purifier unit called Living Air (it makes ozone) to try in the darkroom. I have become somewhat sensitive to Fixer. What about HEPA filter systems? Your opinion or advice on this subject is greatly appreciated. best regards, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I know that Zeiss is sponsoring a "wet" workshop on June 20th during the New Mexico state histology meeting. You can contact Pat Barnes, (505) 262-7000 #7131 for more information. Denise Hardy ARUP
---------- From: Barry, Lilith[SMTP:Lilith.Barry-at-nrc.ca] Sent: Tuesday, May 26, 1998 1:12 PM To: Histonet; microscopy Subject: Microwave techniques course
Is there a hands-on course offered somewhere on microwave techniques (preferably research oriented )? Thank you, Lilith ------------------------------------------- Lilith Ohannessian-Barry NRC, IBS, Ottawa, Ont. K1A 0R6 Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
Hi, We have been using Voxblast by Vaytek Inc. to reconstruct LM deconvolved images and confocal images. I haven't used other programs but we have been very pleased with the intuitive user friendly operation. The movies and animations are fantastic. I'm a user not a vendor.
Bob Underwood Derm Imaging Center Univ. of Wash.
On Sun, 24 May 1998, charles j day wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anyone know of and have any positive experience with any } good sources of 3-D reconstructive software for serial section data? } Shareware or Commercial? If you are a vendor I suppose this list server } would prefer that you respond to my Email directly. I'm also interested } in some secondary manipulative software that can rotate and } change(distort-attenuate or amplify) axis in three(3) or five(5), ie. } two( 2) or more rotational axis. Any other suggestions? } Jeff Day/ "JD" } Mesquite, Texas } Email: wa5ekh-at-juno.com } } _____________________________________________________________________ } You don't need to buy Internet access to use free Internet e-mail. } Get completely free e-mail from Juno at http://www.juno.com } Or call Juno at (800) 654-JUNO [654-5866] }
Thank you to all who responded with ideas for saving partially-polymerized blocks of Spurrs resulting from leaving the out the catalyst.
I am happy to know that there really are others who share my experience, though most volunteered that it was someone else in the lab who made the mistake! The vast majority mentioned A) the blocks would not polymerize by the application of additional heat and B) they could be saved by solubilizing the non-polymerized Spurrs in propylene oxide or acetone, then re-infiltrating.
My story has a conclusion. The batch of Spurrs, to which I neglected the addition of catalyst, was made in the same vessel which contained a few mills of Spurrs from a mixture made earlier in the day, which did have catalyst (someone else in the lab made that batch!). Additional heat (75C) over the long weekend fully polymerized the blocks.
Thank you for your help!
Doug
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
I have noted an increasing use of the Listserver to voice complaints against specific vendors, with a thinly veiled attempt at soliciting replies to justify the posting. This year alone both Oxford and more recently Philips have become the brunt of negative postings. In the recent past other manufacturers (JEOL, EDAX and others) have been similiarly "bashed" by disgruntled users.
I will remind all of you that this forum is for a discussion of microscopy and microanalysis not a venue to list grievances to any particuliar vendor or company. If a specific problem does exist then certainly use this forum to ask a question and , solicit suggestions for its solution. However listing a series of problems which may or may not have been fixed is not only unprofessional it is counter-productive and, I might add, potentially libel especially if the problem was solved by the manufacturer.
Just as we ask vendors not to post advertisements, it is equally improper to use the listserver to impugn a commerical organization regardless of the perceived effect on any individual. The correct procedure would be to contact the National Sales and Service Managers and should that fail the Company CEO together with your Site Procurement Officer. Similiarly it would also be completely improper for a competitor to use any negative complaints posted herein in it's dealings with customers over a product without a complete disclosure of all the details of the problem and solutions which were applied.
We can all document service problems, I for one have had vexing problems with literally every microscope and accessory I use in the lab (and I have instruments from all the major manufacturers' including: Edax, Gatan, JEOL, Hitachi, IonTech, Ortec, Oxford, Philips, SBT, SPI, VG, as well as a number of other companies). In all cases, I have received timely and courteous treatment (including the service organizations of Philips, Oxford, JEOL and Edax). Yes, it is also true that some problems took longer than others to solve but they were all addressed by the respective service organizations.
Realistically, given the volume of Email which I receive daily I do not read every posting to the Listserver and should anyone find a posting which violates the intent of the Listserver I, as SysOp, would appreciate your bringing it to my attention.
As always, should you have a question as to the appropriateness of a posting I will be glad to review it and reply/comment privately on it's suitability before you send it out to the Listserver.
We embedded marsupial testicular and epididymal tissues into LR white resin (made by London Resin Company Limited) for immunogold labelling. The ultra-thin sections have numerous holes which are mainly located on areas without tissues. We though the holes were caused by water moisture, and dehydrated samples with 100% AR Grade ethanol before the embedding. However, holes are still appeared. Could anyone kindly give us some ideas to overcome this problem? Many thanks.
************************************************************************** |Dr Minjie Lin | _-- |\ | Dept of Biological | |biml-at-cc.newcastle.edu.au | / |_\ | Sciences | |Phone:(02)49215707 | \_.--. / | University of Newcastle | |FAX: (02)49216899 | v | Callaghan NSW 2308 | | AUSTRALIA | | | | The CRC for Conservation and Management of Marsupials | | -http://www.newcastle.edu.au/department/bi/birjt/jrcrc/ | **************************************************************************
I've been recommended to use a product called Vertrel XS to clean my detector. I believe it's made by Du Pont. I'm told that it can be used without warming the detector up. Methanol had been previously recommended on a warm detector.
Has anyone used this ? How do you clean yours ? Does anyone know of a source for this product ( preferably UK ) ?
Thanks,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message-----
} membership to the list is dynamic, there are always new members joining. } This informationmay be usefull for others. } } } } Does anyone know of and have any positive experience with any } } good sources of 3-D reconstructive software for serial section data? } } Shareware or Commercial? If you are a vendor I suppose this list server } } would prefer that you respond to my Email directly. I'm also interested } } in some secondary manipulative software that can rotate and } } change(distort-attenuate or amplify) axis in three(3) or five(5), ie. } } two( 2) or more rotational axis. Any other suggestions? } } Jeff Day/ "JD" } } Mesquite, Texas } } Email: wa5ekh-at-juno.com } } } We have a 3D Volume Visualization and Measurement software that performs } what you asked and more. There are several products that can take a stack } of 2D images and create a volume that can be rotated. } } Our product called VoxBlast has many unique features and functions that } allow you to cut, fade, hide, rotate, translate, shade, extract, measure 2D } and 3D, animate, and much more. We also have a module that can take the } images from a scanner and automatically normalize the brightness, cute out } the slides, center them and generate the appropriate volume file for } VoxBlast. I would love to explain all these in detail, but fear that I } would tax the patience of many members. } } If you have an interest in such software, please contact us directly or } visit our web site: WWW.VAYTEK.COM } } Best regards } } Patrick Guerin } Customer Technical Support Engineer } VayTek, Inc. } 305 West Lowe Avenue, Suite 109 } PO Box 732 } Fairfield Iowa 52556-0732 } Tel : 1-515-472-2227 ext. 103 } Fax : 1-515-472-8131 } WWW.VAYTEK.COM } E-mail : pguerin-at-vaytek.com } } Chris MacLean, Ph.D. } VayTek, Inc. } 305 West. Lowe, Suite 109 } P.O. Box 732 } Fairfield, Iowa, 52556-0732 } Tel: 515-472-2227 ext.105 } Fax: 515-472-8131 } E-Mail: cmaclean-at-vaytek.com } Web: www.vaytek.com } -----Original Message----- } From: charles j day {wa5ekh-at-juno.com} } To: Microscopy-at-Sparc5.Microscopy.Com {Microscopy-at-Sparc5.Microscopy.Com} } Date: Monday, May 25, 1998 2:56 AM } Subject: Sources of Software for Reconstruction of Serial Section } Information } } } }
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Hi,
I've been recommended to use a product called Vertrel XS to clean my detector. I believe it's made by Du Pont. I'm told that it can be used without warming the detector up. Methanol had been previously recommended on a warm detector.
Has anyone used this ? How do you clean yours ? Does anyone know of a source for this product ( preferably UK ) ?
Thanks,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message-----
} membership to the list is dynamic, there are always new members joining. } This informationmay be usefull for others. } } } } Does anyone know of and have any positive experience with any } } good sources of 3-D reconstructive software for serial section data? } } Shareware or Commercial? If you are a vendor I suppose this list server } } would prefer that you respond to my Email directly. I'm also interested } } in some secondary manipulative software that can rotate and } } change(distort-attenuate or amplify) axis in three(3) or five(5), ie. } } two( 2) or more rotational axis. Any other suggestions? } } Jeff Day/ "JD" } } Mesquite, Texas } } Email: wa5ekh-at-juno.com } } } We have a 3D Volume Visualization and Measurement software that performs } what you asked and more. There are several products that can take a stack } of 2D images and create a volume that can be rotated. } } Our product called VoxBlast has many unique features and functions that } allow you to cut, fade, hide, rotate, translate, shade, extract, measure 2D
} and 3D, animate, and much more. We also have a module that can take the } images from a scanner and automatically normalize the brightness, cute out } the slides, center them and generate the appropriate volume file for } VoxBlast. I would love to explain all these in detail, but fear that I } would tax the patience of many members. } } If you have an interest in such software, please contact us directly or } visit our web site: WWW.VAYTEK.COM } } Best regards } } Patrick Guerin } Customer Technical Support Engineer } VayTek, Inc. } 305 West Lowe Avenue, Suite 109 } PO Box 732 } Fairfield Iowa 52556-0732 } Tel : 1-515-472-2227 ext. 103 } Fax : 1-515-472-8131 } WWW.VAYTEK.COM } E-mail : pguerin-at-vaytek.com } } Chris MacLean, Ph.D. } VayTek, Inc. } 305 West. Lowe, Suite 109 } P.O. Box 732 } Fairfield, Iowa, 52556-0732 } Tel: 515-472-2227 ext.105 } Fax: 515-472-8131 } E-Mail: cmaclean-at-vaytek.com } Web: www.vaytek.com } -----Original Message----- } From: charles j day {wa5ekh-at-juno.com} } To: Microscopy-at-Sparc5.Microscopy.Com {Microscopy-at-Sparc5.Microscopy.Com} } Date: Monday, May 25, 1998 2:56 AM } Subject: Sources of Software for Reconstruction of Serial Section } Information } } } }
I am interested in knowing if there is any "public domain" macros that will do image analysis of the following using the KS400 software from Zeiss:
1) Particle size (clustered particles of powders) 2) Filter fibers 3) Grain boundaries 4) Fiber cross-sectional area 5) High aspect ratio glass fibers (length) 6) Area fraction
Also, if anyone is fluent in the KS400 language, I would like to chat with you. I have a little programing experience and would like help on how to use the system more.
The detector is a UTW type ( Ultra Thin Window ). I think it is a thin polymer material & definitely not as tough as Be.
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message-----
Dear friends. I have a Microzoom II optical Microscope. This is a very old design, originally by Cambridge Instruments, and until last year, was still being manufactered by Leica. It has the very useful feature of allowing a Very Long Working distance, more than 11 mm with a 50 X objective ( resulting in 1000 X optical magnification with standard 10X eyepiece and 2X Zoom ). I have noticed a problem with the image, which I have also seen in other ( completely different ) microscopes: For samples with large contrast ( for instance a black aborptive spot on an otherewise mirror-like Aluminum coating ), or when looking at a patterened lithography mask ( That's essentially a glass plate with alternate regions transparent or coated with a black emulsion), we see a "ghost" of the image, shifted slightly from it.
This happens Both with transmitted and incident illumination. Of course in the case of transmitted illumination, it can only be seen in those "black-on-glass" samples.
I have tried all sorts of adjustments , and cannot get rid of it ? Is anyone out there familiar with this sort of problem ?
By the way, this is not INTRINSIC to this model of microscope: I have put the same samples under other microscopes of the same model , and have not seen the "ghost". It coould be inherent to the particular series ( the other ones I tested were older ), but that I have no way of knowing.
I will be extremely thankful for help in fixing this. If possible, please respond straight to me:
jrsenna-at-las.inpe.br
Thanks Jose Roberto Senna LAS-INPE CP 515 12201-970 Sao Jose dos Campos SP BRAZIL
Since we make most of these windows I guess I should put my oar in here. We haven't played around with many solvents but have developed a process using methanol. Let the methonal run across the surface of the window, but don't squirt it directly, just the mount. As for other solvents I'll have to take a conservative stance here. The biggest concern is mechanical damage to the window. The slightest brush with a cotton swab tends to puncture it.
The next concern is the aluminum film that seals the polymer from gas and water vapor permeation. Anything that attacks the aluminum should be avoided, since it is only a few hundred atoms thick.
We could do some experiments here if other solvents are deemed necessary.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
I have recently begun embedding tissues in Unicryl for immunostaining for TEM and I have found that while the results are good the ease of sectioning the same block varies greatly from one day to another. I assume this variability is due to changes in temperature and humidity in the lab and was wondering if anyone else had encountered this problem and found a solution.
Thanks in advance,
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-medcor.mcgill.ca
} I will remind all of you that this forum is for a discussion of } microscopy and microanalysis not a venue to list grievances to } any particuliar vendor or company. If a specific problem does } exist then certainly use this forum to ask a question and , } solicit suggestions for its solution. However listing a series of } problems which may or may not have been fixed is not only unprofessional } it is counter-productive and, I might add, potentially libel especially } if the problem was solved by the manufacturer.
I respectfully disagree with this position. Of course, this is your list, so you can do what you want with it. However, a journeyman and even a *master* is affected by the quality of his or her tools. It is not inappropriate or "unprofessional," I think, to discuss those tools and those who vend and service them.
Quite the opposite. I think that if a company has a consistent problem, then it would be an important *service* of this group to provide consumer information. It would be a service not only to the consumers, but also to the companies involved. I know from experience with other companies that they encourage discussion on related newsgroups as one particular method of quality control and monitoring.
I personally don't think that a company has much to fear from a single disgruntled customer. If a company provides good products and excellent service, then those customers who benefit from it will respond to those who trash the company. If a company does *not* provide good products and excellent service, then I don't think that this list should serve the purpose of protecting them from criticism for that lack.
Finally, the threat of libel is, I think, empty. The truth is, as always, an absolute defense against libel. Certainly, folk should not lie about companies they interact with. They should, however, be free to provide information about the quality of those interactions.
If you are going to outlaw criticism of companies, then you must also outlaw praise, and you must outlaw requests for recommendations and responsese to those requests. After all, if only *good* news and positive statementous about a company or product are allowed, then recommendations and reviews in this group are essentially worthless.
I am looking in to Freeze Substitution and embedding in Lowicryl devices and can't find Reichert on the net. Does anyone know where or who they are now?
Also, does anyone have any comments from experience with these devices.
If you want to remove condensed oil from the detector's window, then =
Petroleum Ether or Hexane could be a viable option. I have cleaned two SUTW's with these solvents (one with Petroleum =
Ether and one with Hexane) and the detectors performed very well afterwards.
I do not know if this method is equally effective if the condensate is fr= om Santovac.
The cleaning, as Mark Lund pointed out, has to be done very carefully, without ever exercising any mechanical force on the window itself, e.g. =
do NOT squirt the solvent against the window and do NEVER blow air agains= t it !! The best way (IMHO) is to fill a pipette (10 to 25 ml) with the Petroleum=
Ether, =
hold the pipette close to the upper rim of the window (not touching it !!= !) and let the solvent run down the window by gravity alone (taking the finger off =
the upper opening of the pipette, thereby releasing the liquid to flow). =
This can be done two to three times, and your window should be reasonably= =
clean. The pipette tip should be fairly fine in order to avoid too much pressure =
generated by the flow. You may want to collect the solvent in a petri dish, as it drops off the detector.
Warning: I've done this with the detector at room temeperature! I don't know if =
it works with a cooled detector! =
Also, both Petroleum Ether and Hexane are very volatile and flammable, so= =
take care not to have any open flames, sparks or cigarettes nearby.
Dear Colin, I presume you mean to clean the window of the detector. The Kevex instructions I have for my detector specify clean iso-propanol, trickled very gently down the window, to remove any oil. The detector does not need to be warmed. I have used this several times on both a Be window and thin-window detector. You wrote: } Hi, } } I've been recommended to use a product called Vertrel XS to clean my } detector. I believe it's made by Du Pont. I'm told that it can be used } without warming the detector up. Methanol had been previously recommended } on a warm detector. } } Has anyone used this ? } How do you clean yours ? } Does anyone know of a source for this product ( preferably UK ) ? } } Thanks, } } Colin } } } Colin Reid, } Electron Microscope Unit, } Trinity College Dublin, } Dublin 2, } Ireland.
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
The Reichert ultramicrotome line (and I believe all the other Reichert products as well) now belongs to Leica, and are marketed under the Leica brandname.
We are trying to look at a distribution of fine particles in a polymer using TEM. Grids are coated with the polymer and allowed to dry to a thin film. This works in some cases and is inadequate in others. Does anyone know of a procedure to produce consistent thin films? We do not have access to a microtome. TIA- Mike
Thomas Moninger moninger-at-emiris.iaf.uiowa.edu University of Iowa Central Microscopy Research Facility http://www.uiowa.edu/~cemrf Views expressed are mine.
On Wed, 27 May 1998, Robert Underwood wrote: } } Hello Microscopists, } } I am looking in to Freeze Substitution and embedding in Lowicryl devices } and can't find Reichert on } the net. Does anyone know where or who they are now? } } Also, does anyone have any comments from experience with these devices. } } Bob } Derm Imaging Center } U of Wash. }
Robert Underwood wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Microscopists, } } I am looking in to Freeze Substitution and embedding in Lowicryl devices } and can't find Reichert on } the net. Does anyone know where or who they are now? } } Also, does anyone have any comments from experience with these devices. } } Bob } Derm Imaging Center } U of Wash.
Reichert is now part of Leica. Company address is:
Leica Reichert Division der Aktiengesellschaft Hauptstrasse 219 A-1171 Wien AUSTRIA Tel: (0222) 46 16 41 0 Fax: 46 03 26 Reichert-Jung optical metallographic microscopes.
Henrik -- Henrik Kaker SEM-EDS Laboratory Metal Ravne d.o.o. Koroska c. 14 2390 Ravne Slovenia Tel: +386 602 21 131 Fax: +386 602 20 436 SEM-EDS Lab http://www2.arnes.si/guest/sgszmera1/index.html MVD Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
Hermann Reese wrote ... } } If you want to remove condensed oil from the detector's window, then } Petroleum Ether or Hexane could be a viable option. } ...
I don't know that I'd use these solvents ... like acetone they are stronger than needed and they also have a very large affect of cooling when evaporating ... i.e, thermal contraction may disrupt the window seal. The only reccommended solvent I've seen is etOH while avoiding physical contact with the window ... i.e., rinse without pressure.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
William R. Oliver wrote: } } On Tue, 26 May 1998, Nestor J. Zaluzec wrote: } } } I will remind all of you that this forum is for a discussion of } } microscopy and microanalysis not a venue to list grievances to } } any particuliar vendor or company. If a specific problem does } } exist then certainly use this forum to ask a question and , } } solicit suggestions for its solution. However listing a series of } } problems which may or may not have been fixed is not only unprofessional } } it is counter-productive and, I might add, potentially libel especially } } if the problem was solved by the manufacturer. } } I respectfully disagree with this position. Of course, this is your } list, so you can do what you want with it. However, a journeyman } and even a *master* is affected by the quality of his or her tools. } It is not inappropriate or "unprofessional," I think, to discuss those } tools and those who vend and service them. } } Quite the opposite. I think that if a company has a consistent } problem, then it would be an important *service* of this group } to provide consumer information. It would be a service not only } to the consumers, but also to the companies involved. I know from } experience with other companies that they encourage discussion } on related newsgroups as one particular method of quality control } and monitoring. } } I personally don't think that a company has much to fear from a } single disgruntled customer. If a company provides good products } and excellent service, then those customers who benefit from it } will respond to those who trash the company. If a company does *not* } provide good products and excellent service, then I don't think } that this list should serve the purpose of protecting them } from criticism for that lack. } } Finally, the threat of libel is, I think, empty. The truth is, } as always, an absolute defense against libel. Certainly, folk } should not lie about companies they interact with. They should, } however, be free to provide information about the quality of } those interactions. } } If you are going to outlaw criticism of companies, then you must } also outlaw praise, and you must outlaw requests for recommendations } and responsese to those requests. After all, if only *good* news } and positive statementous about a company or product are allowed, } then recommendations and reviews in this group are essentially } worthless. } } billo
Bravo!
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Hello Hildegard. Go to the homepage for the Center of Cell Imaging at Yale University,
http://info.med.yale.edu/cellimg
This page is made by PhD Paul Webster, and among lots of techniques etc. you'll also find 'Freeze substitution for the poor scientist' Might be worth looking into.
Good luck.
Randi
Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso MH-Breivika N-9037 TROMSO NORWAY
"We are trying to look at a distribution of fine particles in a polymer using TEM. Grids are coated with the polymer and allowed to dry to a thin film. This works in some cases and is inadequate in others. Does anyone know of a procedure to produce consistent thin films? We do not have access to a microtome".
------- Have you tried casting the film in a liquid (e.g. water ?) just as it is done for collodion ?.
you might also try casting on a carbon coated grid or on a glass slide coated with formvar and then floating this in water.
The plastic probably didn't infiltrate into undecalcified bone, so the routine von Kossa should work on sections. Lesley Weston.
On Tue, 26 May 1998, Chism, Sharron wrote:
} Greetings All! } Does anyone have a procedure for the Von Kossa Stain that will } work on Spurr's embedded tissue? We have a bone specimen that has not } been put into decal solution, but has been processed and embedded in } Spurr's. I have cut a "thick" section and have a slide ready to stain, } but need a procedure for plastic embedded tissue. } } Thanks in advance, } } Sharron G. Chism HT (ASCP) } Electron Microscopy Lab } Harris Hospital } Fort Worth, Texas } }
Dear Lilith and all Histonetters- the September 1998 meeting of the National Society of Histotechnology in Salt Lake City Utah will be hosting several microwave workshops including but not limited to-
Saturday Sept 12, Microwave Staining by Chuck Churukian
Sunday Sept 13, Quality Assurance in Microwave-accelerated Procedures by Gary Login
Sunday Sept 13, Antigen Retrieval by Shanrong Shi
Sunday Sept 13, Microwave Processing by Cheryl Crowder and Rick Giberson
Wednesday Sept 16, Antigen Retrieval by K. Wheeler ---------------------------------------------------------------- } Is there a hands-on course offered somewhere on microwave techniques } (preferably research oriented )? } Thank you, } Lilith } -------------------------------------------
Questions in advance regarding the Quality Assurance Workshop can be directed to me at:
Gary R. Login, D.M.D., D.M.Sc. Dept. Pathology Beth Israel Deaconess Medical Center 330 Brookline Avenue Boston, MA 02215
I have experienced holes in sections embedded in LR White and I was told that A. The resin was too old ( It had been around for awhile) and/or 2. I had put the tops on the capsules too soon and the air bubbles had not all risen to the top before I put the capsules into the embedding oven.
} Is there a hands-on course offered somewhere on microwave techniques (preferably research oriented)?
The only such course of which I am aware is a microwave staining class given by Skip Brown. I'm away from my office, but, if anyone is interested, Sonja White at EBS (800-992-9037) can refer you to Skip.
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
Listserver as well as many others places on the Internet is a public place. The owner of the "physical part" of Listserver (like hardware, software) may apply some restrictions on the content of this media (as it happens when we signed for Microscopy Listserver). But I think it is not a good idea to teach everybody what he/she may writes and what may not. The price of "purity of content" on Listserver is too high. Receiving "directives" punishes everybody. For instance, I never posted anything on this Listserver. Why I should hold on my PC a bunch of instructional letters and moreover read those?
I think, everybody who for some reason posted information on this Listserver is responsible for content. If the content is contradicted to agreement, this person should be notifying (personally) about violation of Listserver's rules. Sanctions against violators should be included in the agreement and executed without wild discussion on the Listserver. If we will put such recommendation in practice we will have an well-ordered military-type very plain server without violators and, probably, without subscribers. Is such "order" is Listserver's administration target? If so, I will unsubscribe first.
William R. Oliver wrote: } } On Tue, 26 May 1998, Nestor J. Zaluzec wrote: } } } I will remind all of you that this forum is for a discussion of } } microscopy and microanalysis not a venue to list grievances to } } any particuliar vendor or company. If a specific problem does } } exist then certainly use this forum to ask a question and , } } solicit suggestions for its solution. However listing a series of } } problems which may or may not have been fixed is not only unprofessional } } it is counter-productive and, I might add, potentially libel especially } } if the problem was solved by the manufacturer. } } I respectfully disagree with this position. Of course, this is your } list, so you can do what you want with it. However, a journeyman } and even a *master* is affected by the quality of his or her tools. } It is not inappropriate or "unprofessional," I think, to discuss those } tools and those who vend and service them. } } Quite the opposite. I think that if a company has a consistent } problem, then it would be an important *service* of this group } to provide consumer information. It would be a service not only } to the consumers, but also to the companies involved. I know from } experience with other companies that they encourage discussion } on related newsgroups as one particular method of quality control } and monitoring. } } I personally don't think that a company has much to fear from a } single disgruntled customer. If a company provides good products } and excellent service, then those customers who benefit from it } will respond to those who trash the company. If a company does *not* } provide good products and excellent service, then I don't think } that this list should serve the purpose of protecting them } from criticism for that lack. } } Finally, the threat of libel is, I think, empty. The truth is, } as always, an absolute defense against libel. Certainly, folk } should not lie about companies they interact with. They should, } however, be free to provide information about the quality of } those interactions. } } If you are going to outlaw criticism of companies, then you must } also outlaw praise, and you must outlaw requests for recommendations } and responsese to those requests. After all, if only *good* news } and positive statementous about a company or product are allowed, } then recommendations and reviews in this group are essentially } worthless. } } billo
Dr. Sergey Ryazantsev Department of Biological Chemistry UCLA School of Medicine 10833 Le Conte Avenue Los Angeles, CA 90024-1737 E. mail: sryazant-at-ucla.edu http://www.ben2.ucla.edu/~sryazant
Zeiss/Kontron has a CD called KS Expert with a large number of macros for almost any project. I think there are macros for all of the applications you mentioned.
Thanks Robert A. Carlton Robert.Carlton-at-RP-Rorer.com Tel. 610-454-3949 Fax 610-454-5990
Hermann Reese wrote: ..... : I do not know if this method is equally effective if the condensate is from : Santovac. ..... I have cleaned Be window from Santovac-5 by toluol with good results.
Victor Sidorenko, ANTRON Co. Ltd, scientific service, Moscow, Russia antron-at-space.ru
Hi Jordi Contact your local agent and ask for the KS Expert CD. It has examples of macros for a large number of applications. It is either free of charge, or I got lucky!!
James Wesley-Smith EM Unit University of Natal Durban, South Africa
----------
Dear Fellow Image Analysists:
I am interested in knowing if there is any "public domain" macros that will do image analysis of the following using the KS400 software from Zeiss:
1) Particle size (clustered particles of powders) 2) Filter fibers 3) Grain boundaries 4) Fiber cross-sectional area 5) High aspect ratio glass fibers (length) 6) Area fraction
Also, if anyone is fluent in the KS400 language, I would like to chat with you. I have a little programing experience and would like help on how to use the system more.
Seeing the reference to Reichert and Mager Scientific on the List today prompts me to ask if anyone knows the whereabouts of Jo Ellen. We met in 1986 in Seefeld, Tirol and there's still some pictures on my office wall of us in Innsbruck! Memories of good cryo-courses!
} } I agree with William R. Oliver. } } Listserver as well as many others places on the Internet is a public place. } The owner of the "physical part" of Listserver (like hardware, software) } may apply some restrictions on the content of this media (as it happens } when we signed for Microscopy Listserver). But I think it is not a good } idea to teach everybody what he/she may writes and what may not. The price } of "purity of content" on Listserver is too high. Receiving "directives" } punishes everybody. For instance, I never posted anything on this } Listserver. Why I should hold on my PC a bunch of instructional letters and } moreover read those? } } I think, everybody who for some reason posted information on this } Listserver is responsible for content. If the content is contradicted to } agreement, this person should be notifying (personally) about violation of } Listserver's rules. Sanctions against violators should be included in the } agreement and executed without wild discussion on the Listserver. If we } will put such recommendation in practice we will have an well-ordered } military-type very plain server without violators and, probably, without } subscribers. Is such "order" is Listserver's administration target? If so, } I will unsubscribe first. }
Um, while I thank you for your support, please note that my posting was in disagreement with my perception of Dr. Zaluzec's decision on one point, and *not* a claim that a) the list should not be moderated, or b) that he was doing a bad job.
In fact, I do *not* support this list as a venue for free speech. I *do* support the imposition of controls. I think that Dr. Zaluzec is doing work above and beyond the call of duty in doing this what is, frankly, dirty and boring work for our benefit.
Before I start off a lot of Libertarian flagwaving around here (and, by the way, my bona fides in that area are well established) I think that folk should remember that this is *not* a public forum. Nobody, by limiting speech here, is imposing any governmental restrictions on our inalienable rights of expression and thought. Different venue, different rules. This list is more like a professional panel discussion or other organized meeting -- a church service, committee meeting, whatever.
Accordingly, rules of decorum and subject matter *are* very appropriate. I *don't* want to see a lot of MAKE MONEY FAST posts here. I *don't* want to see advertisements for "hot young nude teens just waiting for my call." Those who really *do* want an unmoderated list are quite within their rights to set one up in competition with this one. Folk will vote with their feet.
I don't want to sound like I'm criticizing you too much, here. I think we are probably very close in our general respect for freedom of expression. However, not *every* venue is a soapbox for random rantings, nor should it be.
Dr. Zaluzek is *right* in saying that folk should not use this list as a place for posts like:
"Hey, Acme Microscope Fumigators were a day late in getting a job done. They are scum-sucking pigs. Their mothers are whores. If you have an Acme store in your town, blow it up!!"
I just don't want to throw the baby out with the bath water here. There should be a place for things like:
"Hey, I'm thinking about having my microscopes de-loused. Any recommendations?"
"Yeah, Beta Bug-B-Gone has always done a good job for me, but Acme has had a habit of being a little slow and expensive, in my experience. I have been de-lousing the microscopes in my lab about twice a year, and over the past 10 years, out price-benefit analysis has favored Beta. Of course, neither company can deal with the ringworm problem that keeps popping up on my objectives"
Lilith, We will have 2 Microwave workshops at the NSH Region I meeting next week in Albany, NY. Skip Brown is presenting Standardization of Histochemical Procedures Through Microwave Technology and Steve Slap is presenting Microwave Applications. Registration is still open, feel free to contact me if you need more information. Mary Georger Astra Arcus USA---------- } From: Hardy, Denise[SMTP:HARDYDD-at-arup-lab.com] } Sent: Tuesday, May 26, 1998 3:54 PM } To: Histonet; microscopy; 'Barry, Lilith' } Subject: RE: Microwave techniques course } } I know that Zeiss is sponsoring a "wet" workshop on June 20th during } the New } Mexico state histology meeting. You can contact Pat Barnes, (505) } 262-7000 } #7131 for more information. } Denise Hardy } ARUP } } } ---------- } From: Barry, Lilith[SMTP:Lilith.Barry-at-nrc.ca] } Sent: Tuesday, May 26, 1998 1:12 PM } To: Histonet; microscopy } Subject: Microwave techniques course } } Is there a hands-on course offered somewhere on microwave } techniques } (preferably research oriented )? } Thank you, } Lilith } ------------------------------------------- } Lilith Ohannessian-Barry } NRC, IBS, } Ottawa, Ont. K1A 0R6 } Tel;613-993-6460 } Fax;613-941-4475 } e-mail; lilith.barry-at-nrc.ca } }
Dear Colleagues I am in the market for an electropolishing unit for TEM specimen preparation. I would appreciate receiving comments/experiences on different units that are out there. Thanks, AG
The Midwest Microscopy and Microanalysis Society will hold its Biologic= al Science meeting on Wednesday, July 1, 1998 at the Blood Center of Wisco= nsin in Milwaukee. The first session will begin at 10:30 am, so start your holiday weekend early by joining us in Milwaukee!
The meeting will focus on the application of various microscopy technol= ogies used to solve biological questions, and here's the slate of speakers an= d their general topics:
R. Albrecht Correlative Microscopy M. Mosesson STEM and Gold-labeling of Fibrinogen J. Harb Transmission Electron Microscopy in Diagnostic Pathology H. Owen Characterization of Pollen Development (molecular biology, EM,=
fluorescence) J. Kinnamon Electronic Image Handling (including ethical aspects of im= age processing) J. Kinnamon Imaging Workshop
Limited space will be available for posters, so here's a great incentiv= e to finish your MSA posters early! Please let me know if you'd like to pre= sent a poster, so we can plan for space needs.
A mailing with further program details and travel information will be s= ent to MMMS members in the next few weeks. If you're not a member and wish to=
attend the meeting, please send me an E-mail message that includes your= name and a mailing or fax address. More detailed information will be sent t= o you towards the end of June.
Hope to see you all in July!
Jane A. Fagerland, Ph.D. President, MMMS jane.a.fagerland-at-abbott.com =
} Hello Hildegard. } Go to the homepage for the Center of Cell Imaging at Yale University, } } http://info.med.yale.edu/cellimg } } This page is made by PhD Paul Webster, and among lots of techniques etc. } you'll also find 'Freeze substitution for the poor scientist' Might be } worth looking into. } } Randi Olsen
You'll also find suggestions in:
Schooley, C. (1986) A poor persons' guide to cryotechnique. Part I EMSA Bulletin 15:98-100; Part II ibid, 16:111-117.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Has anyone out there ever used or purchased a Columbia Instruments optical (aka light) microscope? If so can you contact me directly at rgriffin-at-eng.uab.edu?
I sent this message out to the listserver sometime ago. Here it is again. I hope that it helps.
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I am continuing on the subject below in seeking someones opinion on the difference betweeen the ordinary Struers Tenupol3 and a less known equipment called 550D from South Bay Technology. Are those two comparable and can 550D be used for routine work.....
Old message I am interested in your opinion about electrolyhtic thinning apparatues and their performance. I am planning to buy a jet polishing machine to be used for making thin foils from 3mm disks. The material I am interested in = is High strength Aluminium alloys, steels, stainless steels.
Best wishes Sten Johansson
Link=F6ping University Department of Mechanical Engineering
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Dear Colleagues I am in the market for an electropolishing unit for TEM specimen preparation. I would appreciate receiving comments/experiences on different units that are out there. Thanks, AG
Seeing the reference to Reichert and Mager Scientific on the List today prompts me to ask if anyone knows the whereabouts of Jo Ellen. We met in 1986 in Seefeld, Tirol and there's still some pictures on my office wall of us in Innsbruck! Memories of good cryo-courses!
Keith Ryan Plymouth Marine Lab., UK
We as a purchaser of a Reichert ultramicrotome through Mager Scientific a number of years ago received a courtesy letter from Jo Ellen more than a year ago indicating that she was leaving Mager and moving to Washington State. She did not indicate what she would be doing other than that her husband took a new job out there.
} ..................................................... I think } that Dr. Zaluzec is doing } work above and beyond the call of duty in doing this what is, frankly, } dirty and boring work for our benefit.
} Dr. Zaluzek is *right* in saying that folk should not use this } list as a place for posts like: } } "Hey, Acme Microscope Fumigators were a day late in getting a } job done. They are scum-sucking pigs. Their mothers are whores. If } you have an Acme store in your town, blow it up!!" } } I just don't want to throw the baby out with the bath water } here. There should be a place for things like: } } "Hey, I'm thinking about having my microscopes de-loused. Any recommendations?" } } "Yeah, Beta Bug-B-Gone has always done a good job for me, but Acme } has had a habit of being a little slow and expensive, in my experience. } I have been de-lousing the microscopes in my lab about twice a year, } and over the past 10 years, out price-benefit analysis has favored Beta. } Of course, neither company can deal with the ringworm problem that keeps } popping up on my objectives" }
I agree
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
This generator was mistakenly bought by our lab, so we desperately need to unload it. It is brand new and has never been used and was purchased for $6100.00. Any reasonable offer will not be refused.
Operating Characteristics: Rated Output: 1.0 lbs/ day on air 2.5 lbs/day on oxygen
Rated Concentration: 2.0% by weight on air 3.0% by weight on oxygen
Air Flow: 20 SCFH for air feed 32 SCFH for oxygen feed
Air Pressure: 15 PSIG Dew Point: -60 degree F or lower
If interested email me at this address or call.
Kevin MacKinnon Micro Analytical Laboratories, Inc. 5900 Hollis St., Suite M Emeryville, CA 94608 (510) 653-0824
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
Try contacting Rick Gibberson at Ted Pella 916-243-2200 there is also an 800 number. Rick has given or been part of many Microwave Workshops and has great handouts!
Dear All, I need some input on choosing the right image analysis system for my application. I am doing non-fluorescent, Streptavidin-Biotin-AlkalinePhosphatase-New Fuchsin labeling of cell surface molecules of inflammatory cells in paraffin embedded tissue (Zinc-Tris fixed, Haematoxylin counterstained). I have treated and control inflamed tissue sections and need to compare these for staining intensity, to see if the treatment decreased the number of cell surface markers. I have an image analysis system on trial with a Sony CDX950 3 chip color camera and Northern Eclipse software. I could exchange this camera for a SPOT 12bit CCD, which is much slower, but have higher resolution and possibly truer colors than the Sony analogue camera. Question #1: Is there anybody doing intensity measurements on immunohisto applications I could talk to? #2. Which camera is better for this application? With the Sony camera I am unable to achieve images similar to what I see through the microscope: red and blue becomes less distinguishable, everything turns a bit orange in colour (I have blue and/or heat filters in, but without them it is even worse). Would the SPOT camera be better? #3 How valid is an intensity measurement with New Fuchsin (or DAB)? There are only a few publications out there.... What controls do I need? What colors are better for counterstain - stain (blue-red or blue-green)? #4 What do you measure for intensity: hue, saturation, or value of the pixel?
Any quick help is appreciated, I have left time only until next Tuesday to evaluate the System. Thank you: Leslie
We try to prepare x-sections of cobalt on diamond interfaces=8A not an easy = work!
Besides breaking or crushing the sample, do anybody know a good way to, or reference where to learn how to i) cleave thin chips or tiny pieces of diamond ii) prepare thin slices for ion milling iii) prepare a thin edge or slice (=89 50 - 100 micrometer in thickness) suitable for further FIB thinning? Best regards
Philippe Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
Steve Wintonick asked on the way to measure carbon film thickness without thickness monitor by interference colors on gold or polished brass substrate.
I have a copy of a Balzers report that was given to the TEM users in the mid 1960s: Electron Microscopical Specimen Preparation Techniques from S. Bohler. There are two tables for thickness estimation during coating with carbon and SiO2:
Carbon on an evaporated gold film/test glass (thick enough to behave the "gold color"): approx 150=C5 is orange approx 200=C5 is indigo-red approx 240=C5 is blue approx 350=C5 is blue-green approx 430=C5 is green-silver
SiO2 on an evaporated chromium film/test glass gives a sharp color change: approx 1000=C5 purple approx 1250=C5 blue
Of course, if you place your sample and the test glass at different distances d and D from the source (of small diameter with respect to d and D). the thicknesses will be in the ratio D^2/d^2.
Best regards
Philippe Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
I am posting this message for a colleague who is not a member of the list. Is there anyone on this list who has had experience with staining Bacterial cells with cationized ferritin?
If so, does the stain react with Osmium Tetroxide and to what extent?
In addition does the stain (cationized ferritin) react with Teichoic acid or Teichuronic acid on the bacterial cell surface?
With Best Wishes,
Bill Monroe EM Center Mississippi State University
Liz - This from F.W.D. Rost's "Fluorescence Microscopy Vol 2."
Sulforhodamine G - Excitation=529- Emission=555nm - he also notes a blue (450-490nm) excitation Phloxine B - "Green excitation....absorption max. 546-548nm, shoulder at 510nm" Rose Bengal - "presumably green (546) or UV excitation".
Dave Calvert Eastman Chemical Co. P.O. box 1972 Lincoln Street Kingsport, TN 37664 voice: (423) 229-4943 fax: (423) 229-4558 calvert-at-eastman.com
} -----Original Message----- } From: Liz Nickless [SMTP:E.M.Nickless-at-massey.ac.nz] } Sent: Thursday, May 28, 1998 7:38 PM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: confocal stains } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Does anyone know the excitation and emmision peaks for the following } dyes: } } Sulforhodamine G } Phloxine B } Rose Bengal } } I have a student wanting to try these out on our confocal microscope } for } staining fungal groeth in plants. We have an argon/krypton laser. } } Appreciate any advice. } } Thanks } } Liz
Hi Liz {underline} , {/underline} when I need to know stuff like this I find it helpful to consult the database at http://www.probes.com/probes Other fluorochrome manufacturers surely have the same types of online databases by now? I found the following info. when I searched the names of the three stains you asked about: {underline}
Stain Excitation Emission {/underline}
Sulforhodamine G 529 548
Rose bengal diacetate 313 none?
No luck on Ploxine B though. Per usual, I have no financial interest in the company listed above (I'd have to have some finances first wouldn't I?).
A post-doctoral position will soon available in the Air Pollution laboratory (INRA) in Nancy, France, for a molecular-microscopist biologist. The candidate will study the localization of antioxidant (SOD, GR, and other enzymes) by immunocytochemistry on leaves of different species of trees after fumigation by ozone. Experience in immunolocalization, protein purification and antibodies production is required. The candidate must be highly motivated to succeed Send curriculum vitae, letter of interest, list of articles and references by e-mail before 8 June. Salary will be about 8000FF.
-------------------------------------------- Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
CD burners regularly require whole disk images to be present for burning.
It might be of interest to you, that Retrospect Version 4.0 implements packet writing (specified in the Orange Book , according to the Retrospect manual). Practically, this means that anyone with an Apple Macintosh with an asynchronous SCSI port (PowerPC, Quadra 840AV, and perhaps some more machines) can have their CD burners (supported devices: see http://www.dantz.com) toast away unattended, using all of the Retrospect inclusion/exclusion/compression features.
My setup includes a Powermac 8600, and a Panasonic 7502 CD writer. Running Retrospect is pure bliss for someone relying on backup of large data. A thorough backup strategy has helped me running the machine (and have it up and running 15 minutes after a harddisk crash again) many times more than any other security strategy I tried. Since I started processing larger image data on my machine (scanned macro images, micro images, MR images), this has become a considerably large problem.
Just in case someone wanted to know. Wolf
------------ } In surveys with a total of 24,874 respondents (desktop publishing, web } authors, digital video, animation, etc.), spanning five years of } research, creative professionals reported their actual net profits from } authoring activities by platform. The Macintosh returned the highest } profitability of any platform, over UNIX, Windows, OS2, Sun, and SGI. } Why? Mac-based creative professionals spent more time in creative } activities and less time futzing with "digital administrivia." } } - February 1997, Methodology GISTICS Incorporated, Larkspur, } California
--------------------------------------------------------------------------- Wolf Schweitzer MD, Research Fellow/ Registrar, Melbourne, Australia mailto:wschweitzer-at-access.ch?subject=clicked_on_mail http://www.acess.ch/private-users/wschweitzer/cyberpage.html
As many of you know, I write aggressively about our technologies in a number of trade journals. We have received a number of requests for images which can be used on covers, postcards, etc. Some of these come with "goodies", some just with the opportunity to have one of your images in a prominent location. At the moment, we have a critical need for fluorescence images. If you have something really eye-catching and are willing to share it with the world, please send via 35mm slide or email, with the following information:
Specimen, magnification, dye/probe, filter set specifications and manufacturer
Not microscopy fixation, but there are so many clever people who know about fixing cells on the list, so I thought I would ask.
I often encounter older books with a moldy odor, and these books can prove a severe allergic reaction in me and others. I don't know the toxic material, whether metabolites or spores or something else. Does anyone know a treatment that can remove the odor and kill the fungal cells? A friend has had particl success with charcoal to reduce the odor, but I doubt this would kill remaining cells, which could make more toxins when the weather gets moist in the summer. Thanks for any suggestions.
Leonard R. Corwin Fort Dodge Animal Health Princeton, NJ 08543-0400
A while back someone posted a note saying there was an EM facility in Turkey. Would that person please e-mail me? I have a student who needs to know what type of equipment you have and if she would be able to use it while she was doing research in Turkey. She needs an ESEM because she is working on archealogical samples that can not leave the country and can't be sputter coated.
So PLEASE contact me as soon as you can so we can see if you can help her. Thank you!
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I consulted our Station Mycologist, Nancy Nickerson, and this is what she said:
"His best bet would be to keep the books as dry as possible, because the molds won't grow without moisture, and they aren't likely to produce toxins unless they're actively growing. Certainly he should be working with the books in a well-ventilated area if he has allergy problems. "
Hope this helps.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
I would have sent this info to the original poster directly, but I ran across it after I cleaned out my mailbox. Go to: http://aleph.lib.ohio-state.edu/www/hasafran/hasafran480.html to read about several methods of dealing with mold in books. If you don't have web access I'll e-mail it upon request.
I am interested in your comment that "you need an ESEM because you can no= t coat the specimen". True in many cases specimens will charge at "normal"=
operating kilovoltages and beam currents. Have you tried the gentle approach, "low charge techniques", with the type of specimen in question= =2E
1. Lower kV, try 2 to 5kV for example on an instrument up to 10 year= s old, on a newer instrument you could probably get down to 0.5 kV, it all depends on the resolution that you require. 2. Lower the probe size, hence beam current, less electrons means a greater possibility of those you are using getting away to earth. 3. Change the WD (and tilt), in different instruments different WD (and tilt) give different signals to the ET detector there will always be= a position of minimum charge effect.
We are often in the situation that you describe and as ESEM are few and f= ar between we use low charge techniques on whatever instrument we happen to have. All this is even better with LaB6 or a FEG system.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
This was discussed in the Safety newsgroup quite some time ago - perhaps early last year. There were a lot of ideas put forward from professionals in the book business. Their archives are at
http://siri.org/mail and at http://list.uvm.edu/archives/safety.html
If you can't find the discussion and would like info on subscribing, let me know. Diana.
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
This was discussed in the Safety newsgroup quite some time ago - perhaps early last year. There were a lot of ideas put forward from professionals in the book business. Their archives are at
http://siri.org/mail and at http://list.uvm.edu/archives/safety.html
If you can't find the discussion and would like info on subscribing, let me know. Diana.
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
Anybody out there who can tell me the email address of the University of Moscow, Russia. Dept of Electron optics or maybe physics. Any assistance appreciated Peter Earl Toronto,Canada
Just a quick note on gold, carbon thickness monitors. The gold can be= =20 deposited on the smooth side of Aluminum foil or we use 3-4 mil smoo= th=20 thin sheet for stiffness. It can then be cut to a convenient size ea= sily. ---------- =46rom: philippe.buffat-at-epfl.ch To: microscopy-at-Sparc5.Microscopy.Com; daniele.Laub-at-epfl.ch; =20 fabienne.bobard-at-epfl.ch; gaston.peter-at-epfl.ch; Bernard.Garoni-at-epfl= .ch
Steve Wintonick asked on the way to measure carbon film thickness wit= hout thickness monitor by interference colors on gold or polished brass substrate.
I have a copy of a Balzers report that was given to the TEM users in = the mid 1960s: Electron Microscopical Specimen Preparation Techniques fro= m S. Bohler. There are two tables for thickness estimation during coating with car= bon and SiO2:
Carbon on an evaporated gold film/test glass (thick enough to behave = the "gold color"): approx 150=C5 is orange approx 200=C5 is indigo-red approx 240=C5 is blue approx 350=C5 is blue-green approx 430=C5 is green-silver
SiO2 on an evaporated chromium film/test glass gives a sharp color ch= ange: approx 1000=C5 purple approx 1250=C5 blue
Of course, if you place your sample and the test glass at different distances d and D from the source (of small diameter with respect to = d and D). the thicknesses will be in the ratio D^2/d^2.
Best regards
Philippe Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time= ) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 __________________________= _
This is a reminder that the Early Registration Deadline for Microscopy and Microanalysis '98 is approaching. This deadline has been extended to June 15, 1998, so be sure to get your registration in by that date to save money! Registration forms are available in the Call for Papers, or on the MSA Web site, www.MSA.Microscopy.com. Information on the Technical Program is also available on the Web site. MSA members will also be receiving the Program Book and a Meeting Information Pamphlet in the mail within the next week.
All indications from exhibit sales and early registration are that Microscopy and Microanalysis '98 will match or exceed the successes of M&M'96 (Minneapolis) and M&M'97 (Cleveland). Hotels in Atlanta are filling up quickly, so be sure to also get your reservations in (hotel reservation forms are also available in the Call for Papers, the Meeting Information Pamphlet, and the Web site).
For general information about the meeting, contact the Meeting Manager at 708-361-6000, or at msa-at-tradeshownet.com
A colleague has asked me to inquire if any listmembers have an SEM that will accommodate a small object about 3.5 inches thick (large Z capability). The colleague works in the Kansas City area, but the object can travel.
Thank you.
James Martin Analytical Services and Research, Williamstown Art Conservation Center Research Scientist, Williams College tel: 413.458-5741 fax: 413.458-2314
The Silicon Electronics Research Lab at Bell Labs, Murray Hill, New Jersey, is looking for a TEM technician. Our research interest is in analytical and High Resolution TEM, mainly of silicon based devices and structures.
In our lab, we have a JEOL 4000 EX and a Philips EM420, and will soon purchase an FEG machine. In addition, we have a Micrion FIB. For sample prep, we have the mainly Gatan equipment (Dual Mill, PIPS).
Recently, the TECHNICIAN who looked over the TEMs and the sample preparation facilities retired. We would like to fill this position again.
We are looking for a person who might be interested in helping us running the TEMs and the sample preparation, preferably someone who is able to do TEM specimen preparation and some investigations on his own? (Or who would be willing to learn.) The work would be rather technical than a scientific ("more like tinkering with equipment").
Interested persons may reply by email to frieder-at-lucent.com.
Hi there, a quick question, has anyone had any experience with mounting a Gatan 622 image intensified TV camera on to a JEOL 4000EX (or FX) that already has a Gatan Imaging Filter installed? We have just installed the GIF and also want to use a TV rate camera at lower mag. We dont really care if it has to be mounted off axis or at a weird angle (we do need to get close enough to the microscope to operate it!!). Thanks.
If no one has experience and/pr it is not possible, does anyone want to buy our 622 TV camera?
Note new Area Code (734)
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Go to our Links. Click on references and find international universities (or use browser's search function for "international" or "Demello". That will give you several homepages for Moscow universities. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} } Anybody out there who can tell me the email address of the University of } Moscow, Russia. } Dept of Electron optics or maybe physics. } Any assistance appreciated } Peter Earl } Toronto,Canada } } }
Please bring this technical position opening to the attention of your colleagues. We will accept applications until the position is filled. Thanks!
Bob Summers Director, Confocal Microscopy and 3D Imaging Laboratory SUNY Buffalo ------------------------------------------
SENIOR RESEARCH SUPPORT SPECIALIST: IMAGING LABORATORY State University of New York at Buffalo
DESCRIPTION OF DUTIES: Manage the daily activities of a campus wide biological imaging resource and associated satellite facilities operated by the School of Medicine and Biomedical Sciences. General areas of usage of the laboratory relate to the preparation of biological images for research, publication and instruction. Duties include instruction of faculty, students and technical personnel in the use of digital and analog imaging equipment including confocal microscopes, molecular imagers and scanners, 35mm slide making equipment, video capture and production equipment, photography equipment and printers. Must be familiar with research standards and practices to maintain quality control and advise users on appropriate protocols. Responsible for daily and long-term maintenance and calibration of equipment. Must hire, train and supervise graduate assistants and assign them to assist users of the imaging resource. Develops and manages budget, maintains financial records of the laboratory and collects user fees from users of the laboratory. Should work closely with Director and Faculty Advisory Committee to establish needs, set priorities and develop policies within the laboratory.
QUALIFICATIONS: MS or MA in Biological Sciences. Research experience that includes microscopic and macroscopic biological imaging, graphics illustration and photography. Preparation of biological specimens for microscopy. Experience in use of computer hardware to include Apple, PC and Unix platforms and operating systems, scanners, imagers and digital microscopes. We desire experience with computer software including PhotoShop, Corel Draw, Canvas, PowerPoint, Persuasion, Desktop Publishing and Word Processing Programs. Experience in installation of software and maintenance of computer hardware. Must have experience with using computer networks. Personal skills should include working with large number of researchers and/or educators to expeditiously handle work requests and to attain customer satisfaction.
SALARY commensurate with qualifications. SUNY Buffalo is an EOAA Employer
FOR INFORMATION OR TO APPLY CONTACT: Dr. Robert G. Summers, 716.829.2911 (phone) or fax credentials to 716.829.2915 or email to: summers-at-confocalsgi.med.buffalo.edu
I'm looking for a software (freeware or shareware) for diffraction patterns analysis.
Thanks !
******************************************** * Paulo C. Soares Jr. * * LaMaV - Laboratory of Vitreous Materials * * Federal University of Sao Carlos * * Sao Carlos - Brazil * ********************************************
There will be a Photomicrograph Exhibit held in conjunction with the 14th International Congress on Electron Microscopy in Cancun this August 31 - September 4. If you are attending the meeting, please consider bringing up to 3 micrographs for inclusion in a Photomicrograph Exhibit. From this exhibit, images will be selected for inclusion in a post conference publication. See details below and if you have questions contact either the meeting headquarters in Mexico City (info below) or myself.
Photomicrograph Exhibition
The 14th ICEM will include a micrograph exhibition. The organizers' intentions are to provide a showcase for outstanding micrographs displaying a combination of art and science.
The exhibit is open to all forms of microscopic imaging. A panel of judges will select entries based on artistic merit for inclusion in a post-congress publication of images. Submitted micrographs much be accompanied by a brief description, not only of their scientific content, but also of their technical aspects. If images have been digitally processed or altered, the digital processing should be described as well. All submitted micrographs will be displayed although entries not regarded by the judges as artistically significant will be excluded from the publication of images. Please read the following rules carefully:
Only registered attendees at the 14th ICEM are eligible to submit micrographs.
Any individual may submit up to three micrographs.
Entries must be 27.5 cm x 35.0 cm, may be mounted vertically or horizontally, and must be affixed to a stiff lightweight support, such as 10 mm foam board. Micrographs may either be flush mounted or have borders so long as the overall dimensions of the entry are 27.5 cm x 35.0 cm.
Entries must be brought to the meeting and mounted on the display boards by the entrant or his/her delegate. Velcro will be provided. Entries must be mounted between 12:00 and 17:00 on Monday, August 31st and those micrographs not selected for publication must be removed between 12:00 and 15:00 on Friday, September 4th. Micrographs remaining on the display boards after then will be discarded. Micrographs selected for publication will not be returned to the entrant and will become the property of the 14th ICEM.
Selected micrographs will be incorporated into a post-congress publication of images. Those selected for inclusion will be announced during the Thursday evening reception.
To enter: Send your name, address, telephone and fax numbers, and email address plus a description of 200 words or less for each entered micrograph (maximum of 3) to: Secretariat 14th International Congress on Electron Microscopy Amsterdam 46-202 Col. Hipodromo Condesa C.P. 06100, Mexico, D.F. MEXICO Phone: (525) 553-4507; Fax (525) 553-4500 email: icem-at-icem.inin.mx
Entry information must be received by August 1, 1998. Entries will be acknowledged promptly. Do not send micrographs, you must bring them or have them brought to the Meeting.
Meeting Office staff will print your description(s) in standard form and prepare them to be mounted along with your micrograph(s) at the Meeting.
___________________________________________________________________________ Barbara Reine, Botany Dept. Box 351330 Univ. of Washington, Seattle, WA 98195-1330 e-mail: reine-at-u.washington.edu; Phone: (206) 543-1955; Fax: (206) 543-3262 ____________________________________________________________________________
Hi, Does any of the listers know any ER specific or Golgi specific antibodies for immuno-gold or confocal staining? Thanks.
Li-Tzu Li Dept. Medical Technology College of Medicine National Taiwan University Taipei, Taiwan Phone: 886-2-2397-0800 ext 6907 Fax: 886-2-2381-7083 Email:ltl-at-ha.mc.ntu.edu.tw
If you have a reasonably quick web connection, why don't you try EMS On-Line at http://cimewww.epfl.ch/ . This does loads of things which also includes image simulation of TEM images. I have found it very useful for indexing SAD patterns recently.
Hope this helps
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
TECHNICAL ASSISTANT: LIGHT AND ELECTRON MICROSCOPY AND X-RAY SPECTROMETRY
This half-time position will oversee the light and electron microscopy facility including scanning and transmission electron microscopes, a laser scanning confocal microscope, an energy dispersive X-ray spectrometer, digital image analysis equipment, diverse sample preparation equipment, and two darkrooms. The facility serves Biology and Geology, as well as Art, Astronomy, Biochemistry, and Neuroscience faculty, research students and teaching labs.
Responsibilities: oversee the day-to-day operation of the facility, including routine maintenance and troubleshooting of instrumentation, ordering supplies, and general laboratory and darkroom maintenance; provide training to faculty and students in specimen preparation, proper use of the various microscopes and detectors, digital image processing, and traditional darkroom techniques; prepare laboratories for specific courses; maintain microscope inventories; assist with other designated instructional tasks. The applicant must have excellent communication skills, provide expertise to people of diverse skills and research interests, be able to set priorities and work without direct supervision, and bring enthusiasm to the job. The position carries the possibility of both additional teaching responsibilities with added compensation and use of the facility for research.
Requirements: Master's degree in science with considerable experience in the areas of electron and confocal microscopy or X-ray microanalysis. Computer literacy (preferably in both the Macintosh and Windows environment) is desirable. Annual starting salary is $17,500 and includes a comprehensive benefits package.
Review of applications will begin July 1 and continue until the position is filled. Forward a letter of application, resume and the names and telephone numbers of three professional references to: Richard Briggs, Department of Biological Sciences, Smith College, Northampton, MA 01063. Smith College is an equal opportunity employer encouraging excellence through diversity.
Hi all. Apologise for the lack of chit chat. I too have been struck by the = influenza virus doing the rounds.=20
Again as part of the ongoing promotion of MSSA and microscopy in SA. We = have a small portable Hitachi SEM available to us which could be used to = do demo's at schools. It is a very old system and imaging is not the best, but it should be = interesting for school students and teachers to see a working SEM.
The best way to promote microscopy and the E.M.unit at universities, is = to involve both, and so by letting a biology student from a university = in the region, for example, demo the SEM, would be the ideal way of = going about having such a visit to a school. The SEM is here in our workshops in JHB, but we can make a plan to get = it around the country. To connect it up takes a 15A 220v Ac mains plug and a water supply. It = will fit on one standard table and fit in the boot of a car.
So if any body would like to assist with the organising of such a visit, = please let me know.
At this stage I would need to know who is interested and which schools = are interested, then we can start to arrange for dates.
Look forward to your responses.
P.S. this system is not for sale.=20 Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
One of the "classic" ER-staining antibodies seems to be anti-BiP (immunoglobulin binding protein).
See for more information and additional references the following paper:
Terasaki M, Reese TS. 1992. Characterization of endoplasmic reticulum by co-localization of BiP and dicarbanocyanine dyes. J. Cell Sci. 101, 315-322.
Scott Gens gens-at-biodec.wustl.edu
On Tue, 2 Jun 1998, Li-Tzu Li wrote: } Hi, } Does any of the listers know any ER specific or Golgi specific } antibodies for immuno-gold or confocal staining? Thanks.
with SMTP (Eudora Internet Mail Server 1.2); Tue, 2 Jun 1998 11:33:09 -0500 Message-Id: {v0151010cb199d6d02266-at-[137.99.40.57]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Does anyone know where I can get a product called "Coat-quick G"? It is mentioned in a protocol for coating grids which are to be subjected to resin etching and immuno labelling. I assume it is to aid adhesion of the sections to the grid during these procedures. If you do not have a source for this product, perhaps you know of a similar one. Thanks.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
--------------------------------------------------------- The Electron Microscopy Laboratory at New Mexico State University has an open position for an Electron Microscopy Specialist. The laboratory is a multi-user facility that provides TEM, SEM, and light microscopy services for the university research community and a few external organizations.. Qualifications: MS Degree minimum, Ph.D. Degree desirable, with at least three years of electron microscopy experience. The preferred candidate will be skilled in transmission electron microscopy and immunohistochemical techniques as applied to the study of plant and animal (nervous system; cell culture) cells. Knowledge of fluorescence microscopy, SEM, digital image analysis, and the use of networks in computing are assets. He or she also should be experienced at teaching microscopy fundamentals and in managing a multiuser teaching and research laboratory. The ability to work well with research faculty, other EML staff and students is essential. Duties: operation and routine maintenance of microscopes and associated equipment, sample preparation, coordination of projects, billing, course instruction, supervision of student research projects, some educational outreach activities, technical support for EML components of sponsored grants, training of laboratory users. Salary: $33,544-$40,500 DOE plus 26% fringe benefits. Review of applications will begin June 19 and continue until the position is filled. Applications should include a resume, a letter of interest and three letters of reference.
Apply to: Dr. Reed Dasenbrock Associate Dean/Director Arts and Sciences Research Center New Mexico State University MSC RC, Box 30001 Las Cruces, NM 88003 rdasenbr-at-nmsu.edu
For more information about the NMSU EML access: http://www.nmsu.edu/Research/artsci/public_html/eml/index.html#staff
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Position Available: 2-year postdoctoral fellowship in plant pathology employing histology and electron microscopy.
A research position is available for a postdoctoral fellow to work in cereal pathology under the NSERC Visiting Fellowship program.
The research will be conducted in the Crop Science section at the Agriculture and Agri-Food Canada Research Centre in Lethbridge, Alberta, Canada. The successful applicant for this position would be working as part of a research team with Dr. R. L. Conner (cereal pathologist) and E.G. Kokko (electron microscopist).
The study will compare the development and spread of different black point fungi in resistant and susceptible wheats. The investigation will involve a histological examination of wheat tissue using light and electron microscopy. The overall objective is to clarify the factors involved in grain discolouration caused by this disease and the mechanisms controlling disease resistance.
The candidate for this position must have a PhD in Botany, Plant Science or Plant Pathology from a recognized University. Previous experience in light microscopy as demonstrated in research publications is essential. A strong background in plant biochemistry is also required. Familiarity with procedures used in scanning and transmission electron microscopy would also be an asset.
The appointment is for a two year period and salary will be according to current rates for the NSERC Visiting Fellowship Program. Current starting salary is $35,184.00 (Canadian dollars).
The Lethbridge Research Centre is looking for candidates for this position. Applicants should submit a brief resume describing their education background, research activities, and a list of publications along with at least two professional references. Only those who are considered qualified will be given further consideration and contacted personally.
For further details, contact:
Eric Kokko Agriculture and Agri-Food Canada Lethbridge Research Centre P.O. Box 3000, Lethbridge, Alberta CANADA T1J 4B1
Dr. Cantino, Coat Quick "G" is sold by Electron Microscopy Sciences, Fort Washington, PA, 800-523-5874 (EMS Catalog XII, p.68). -------------------------------------------------------- Winston W Wiggins, Supr. 6/2/98 3:41:06 PM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org --------------------------------------------------------
Fellow Microscopists, Recently our lab purchased a second TEM for asbestos analysis. Can anyone give me some advice on screen design for a JEOL 100CX specifically used for asbestos analysis? Currently we are using a Hitachi 7000 with an (11cm. max) concentric circle design. The only problem I find with this is trying to determine the width/diameter of a fiber for a weight % analysis. The smallest division is 0.25 cm., which makes it very difficult to measure individual fibers at a mag of 20,000X. Any input would be greatly appreciated.
Cheers,
Kevin MacKinnon Micro Analytical Laboratories, Inc. Emeryville, CA 94608 (510) 653-0824
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
Hello, my name is Jeff Pierce. I am a microbiologist working for Zeneca in Wilmington, Delaware and have been given an assignment to photograph ( by SEM ) basic pictures of bacteria, fungi and yeasts to be then colorized and used as photographs in technical literature. ( Zeneca manufactures specialty biocides for industrial applications such as coatings, polymers, pigments, etc. ) Anyway, I did a lot of TEM work at Penn State and only really dabbled in SEM. To the question at hand: does anyone have any neat ideas regarding making interesting SEM pictures of bacteria, yeats or fungi? I used to simply filter cultures thru a nucleopore membrane and photograph the membrane with the bugs trapped on the surface. However, the background ( the filter ) takes away from the bacteria. Does anyone have any suggestions that may help? Also, where can I find a current SEM dehydration/sample prep technique for these types of specimens? Any help/comments/suggestions would be greatly appreciated. Thanks in advance, -Jeff Pierce.
I got a few responses to my questions on the "megapixel" consumer digital cameras vs. scientific digital cameras. Many issues are still fuzzy, but since there was some interest in the responses I have paraphrased them below.
But first - would the person from Kodak who responded please re-send your E-mail? (I accidentally deleted it)
RESPONSES:
1. The grade of CCD chip used can differ greatly between the lower and higher priced models. They are graded on their defects (either cluster or column/row defects), which affect the ability to record data. Chips with few or no defects are rare and reserved for the highest scientific grade cameras, from manufacturers like Princeton Instruments and Xilliger [and possibly others not mentioned by the respondent]. A single-chip B/W camera in this grade might cost $40k or more for 1kx1k resolution. The application dictates the need - intensity and color analyses being one application where high quality chips would be desired. The respondent also mentions that Princeton Instruments (no connection) has further information on chip defects.
2. Some references for reviews of digital cameras: Popular Photography (Dec. 1997), PEI - Photo Electronic Imaging (May 1998) www.peimag.com, PC Magazine (February 1998). This person chose the Olympus D-600L after receiving quotes from National Graphics Supply 800-223-7130, B&H 800-947-5516, and Wolf Camera.
3. A recommendation for the MicroLumina (couldn't tell if from a vendor or not??) with 3kx2k resolution, 36 bit color, and standard Nikon 35mm lenses or microscope adapter. Web page http://www.electroimage.com
4. A response from DVC company recommending their web site Q&A/FAQ section on digital cameras at http://members.aol.com/dvcco Noted that camera differences have to do with dynamic range/ bit depth and speed.
5. Responses from other vendors who simply left their phone numbers and offered to discuss the issue.
If anyone wants specific details or vendor phone numbers please E-mial me privately.
Karen
} } ORIGINAL QUESTION: } ------------------------------------------------- } To the digital die-hards out there: } } I know there have been MANY threads lately on digital cameras, which } have been very informative. But there is such a boom going on out } there! Does anyone have a good handle on the differences between the } "megapixel" models which are marketed for consumer/professional } photography markets vs. the scientific models discussed on this list?? } } Specifically, a colleague recently challenged me as to why I would want } to "waste" $6000 on a Minolta RD-175 when I could get even better } resolution from some of the Olympus, Kodak, Konica, Canon, etc etc. } products out there for $1000 or less. After some web site searching, it } seems like there might be an issue of true color representation (1CCD } vs. 3CCD's) and perhaps lens mounts, auto-white balance w/o manual } override, compression, or other things. But I'm not sure per particular } model. Anyone want to get into some of these details?? } } NOTE: my needs are for Both hand-held close up work and possibly } microscope-mount, to be used in subsequent image analysis of colors & } densities as well as morphology. Low-light sensitivity not essential. } } Some of the suggested models were: } } High End: } Minolta RD-175 } Kodak DCS 420, 410, and 460 [anyone know a good vendor in Minnesota?] } Polaroid DMC2000 } } Lower End or Unknown: } Sony DKCST5 [again, any vendors in Twin Cities area?] } Kodak DC120 or MDS120 } Konica Q-M100 } Olympus D-600L } Canon EOS DCS 1 } Pixera Professional [I have this] } many others I haven't looked up yet } } I hate to beat a dead horse, but this is more like cutting up a } starfish. The issues just seem to multiply! } Any input appreciated, } Karen
-- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
FEI Company has the following position available at their Mahwah, NJ location:
FEI Company Job Opportunity Mahwah, NJ
National Technical Support Specialist - SEM
Summary: Provide technical support to ensure customer satisfaction and profitability. Provide first line support to both Philips customer service engineers and customers via the technical assistance center.
Duties and Responsibilities: * Oversees policies and processes which motivate and develop talent to provide technical support with the product group. * Acts as a liaison between supply center and field personnel to resolve technical problems. * Provide means of Technical Assistance Center (TAC) support to assist in customer telephone fix or call avoidance. * Evaluates field Customer Service Engineer's (CSE) for training requirements and makes recommendations to National Customer Service Manager. * Writes service bulletins to field as needed. Interfaces with bulletin boards as needed. * Prepares and conducts service training courses as required. * Reviews and evaluates all new products and documentation. Monitors new product installation. * Develops corrective action plans to resolve complex/long-term product reliability problems through use of MFG-Pro data analysis. * Assists to completion all field installations, upgrades and escalations via telephone daily. * Establishes initial stock levels for spare parts. * Provides technical assistance via telephone to all X-Ray personnel including review of orders as required. * Provides on-site technical assistance to CSE's when required. * Ensures the adherence to company policy, suitability of work instructions, implementation of corrective action and maintenance/accessibility/retention of quality documentation. * Ensure that ISO 9000 directives that pertain to this area of responsibility are fully complied with.
Required Knowledge, Skills and Experience: * BS/BA or AAS Electronic Technology or equivalent. * Must have excellent mechanical and electronic aptitude * 5+ years experience with SEM's (scanning electron microscopes) * Must possess excellent communication and problem solving skills * Must have a working knowledge of software operating system.
Interested candidates should FAX or email resume to Lisa Olivia FAX: 503.640.7509 email: lolivia-at-feico.com
Lisa Olivia Recruiter FEI Company PH) 503.844.2601 FAX) 503.640.7509 email: lolivia-at-feico.com
#03171-Lab Specialist Senior - CISAT Assists in all aspects of procedures required for the prepapration of tissue for immunohistochemical analysis (whole mounts as well as sections), fluorescent and light microscopy, transmission and/or scanning electron microscopy. Competitive candidates must possess knowledge of techniques for immunological staining, sectioning, mounting tissue, evaluating sections and taking and developing publicaiton quality photomicrographs. Applicant must possess demonstrated ability to operate: ultramicrotome, vibratome, cryostat, and other TEM/SEM equipment. Applicant will be responsible for maintenance of specimen records, chemical supplies, reagents, etc in the histological laboratory and for independently modifying lab procedures to obtain optimal results. Interpretation of findings and ability to direct the work of others also necessary. Four year degree in Biology or related field supplemented by post-graduate experience in microbiology and/or neuroscience preferred. Salary range: $24,337-37,995.
Send resume and all correspondence to:
Brenda M. Ryals, Ph.D Professor Communication Sciences and Disorders James Madison University Harrisonburg, VA 22807 ryalsbm-at-jmu.edu
We are looking for a used TEM (in reasonable working condition) that anyone might have for sale or donation. The machine will be used for routine imaging. Please contact me offline if you have any information.
Thanks,
Mohan Kalyanaraman
Sr. Staff Material Scientist Mobil Technology Company PO Box 480 Paulsboro, NJ 08066 mohan_kalyanaraman-at-email.mobil.com
This message is in MIME format. The first part should be readable text, while the remaining parts are likely unreadable without MIME-aware tools. Send mail to mime-at-docserver.cac.washington.edu for more info.
I, Dr Yogis Naidoo a research officer in the Electron Microscope Unit at University of Durban-Westville, South Africa am interested in a post doctoral research position. I will be able to provide my own funding. My field of specialisation is ultrastructure and cytochemistry salt glands of halophtyic plants. My current project focuses on investigating the ultrastructure and cytochemistry of salt glands of halophytic grasses along the South Farican coastline. I am also interested in glands of other related plants. I will be willing to give lectures on plant pathology.
Thank you Yours sincerely Y. Naidoo Electron Microscope Unit University of Durban-Westville Private Bag X54001 Durban 4000 Tel: (031) 2044360 Fax: (031) 2044809
Would anyone be able to recommend us on what diamond knife angle would be a better choice for cutting polymer (polymer-metal/glass fiber composite)? 45 degree or 35 degree or doesn't matter?
I was told by Diatome that 35 degree knife would be a better choice to cut polymers, i.e., it reduces the curling problem. But we are currently using the 45 degree knife. Would it be wise to resharpen the one we have or buy a new one that is 35 degree?
"Analytical Electron Microscopy of Oxynitride Glass Ceramics"
A post-doctoral position is available in the Division for Microscopy= and Microanalysis at Chalmers University of Technology within the framework= of the Training and Mobility of Researchers (TMR) Programme. Qualified candidates should have a Ph.D. in materials science, physics or chemistry, and a documented background in analytical electron microscopy. It= will be considered an advantage if he or she has some experimental experience= of any of the following techniques: electron energy loss spectroscopy= (EELS), convergent beam electron diffraction (CBED) or high resolution lattice imaging. The applicant must be a national of a Member State of the European Community. The starting date of the appointment may be= discussed, and the appointment may last up to 16 months.
The position is funded by the TMR networks research project "Structure= and Properties of New M-Si-Al-O-N Oxynitride Glass Ceramics (M=3DY,Ln)"= which is a collaboration between seven partners in the United Kingdom, Ireland, =46rance, Sweden and Belgium. The collaboration within the established network will give the post-doctoral fellow good contacts with a number= of research groups in Europe and a good knowledge of a variety of preparative / analytical / property measurement techniques relevant to the evaluation of new ceramic materials as well as other materials outside the field= of ceramics.
The aim of the research project is to determine crystal structures= and properties of currently uncharacterised new glass-ceramic phases= in yttrium and rare earth sialon systems. These materials are potential refractory grain boundary phases in Si3N4-based ceramics, and have an interest= also as refractory surface coatings (glazes) and as interface materials in nitride-oxide and nitride-metal joints.
The analytical transmission electron microscopy work carried out= at Chalmers will focus on the chemistry and structure of glasses and= their crystallisation products, and on microstructural development during nucleation and growth processes. Microanalytical work on glass ceramics subjected to creep testing and oxidation / corrosion experiments= may also be carried out.
The major part of the microscopy work will be carried out on a Philips CM200 Supertwin transmission electron microscope (TEM) with a field emission gun (FEG) and surrounding interactive instrumentation. = The FEG TEM is equipped with the Gatan imaging filter (GIF) which produces= energy filtered electron images and diffraction patterns as well as electron energy loss spectra, a Gatan off-axis CCD camera for high resolution imaging and a Link Isis energy dispersive X-ray (EDX) system for qualitative and quantitative elemental analysis including the light elements. There are also other electron microscopes in the laboratory, e.g. a Jeol 2000-FX TEM/STEM/SEM and a CamScan SEM.
=46or further details, please contact: Associate Professor Lena Falk, Department of Experimental Physics, Chalmers University of Technology, SE-412 96 G=F6teborg, Sweden e-mail: lklfalk-at-fy.chalmers.se; fax: +46 31 772 3224; phone: +46= 31 772 3321
Does anyone have a really goood fixation for lm of very hard seeds. We would like to section buckthorn seeds.
Debbie Lietz Electron Microscopy Suite Department of Biology Trent University Peterborough, Ontario K9J 7B8 Telephone: (705)748-1486 Fax: (705) 748-1205 email: dlietz-at-trentu.ca
There has been a lot of discussion recently about substituting one DP oil for another. Here is a summary of the properties of some of the common brands of oil: MM VP BT HV
MM = Avg molecular mass; VP = vapor pressure at 20 C (Pa); BT = boiling temperature (deg C) at 100 Pa (typical DP boiler pressure); HV = heat of vaporization (kJ/mole)
These values are probably not accurate to a high degree, because it is very difficult to measure the physical properties of very viscous, low vapor pressure fluids such as these; however, they should be good enough to give you an idea of the relative characteristics of the various fluids.
In general, if you are going to substitute one oil for another, the two should be as similar as possible with regard to boiling temperature and heat of vaporization. Rough values of boiling temps are given above; however, I could not obtain values for the heats of vaporization that I could feel confident enough about to publish in my book. The values I did come up with are given above, and are only VERY ROUGH. Most manufacturers do not give heat of vaporization data, even though it is a very important property. It is possible to calculate it from data giving variation in vapor pressure with temperature (see VM in EM p 181) but the vapor pressure data are usually not precise enough to yield results one can really trust.
More details on diffusion pump fluids can be found in Sect. 5.4 of my book 'Vacuum Methods in Electron Microscopy' (http://www.2spi.com/catalog/books/book48.html ; http://www.bookshop.co.uk/portland/)
Incidentally; concerning the character of Lion-S Oil, two weeks ago I wrote to the Lion Oil Company requesting information about this oil, but have not yet received a reply.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
As I have mentioned on several previous occasions, it is quite possible to clean most parts of the vacuum systems of EMs and other common laboratory apparatus WITHOUT the use of organic solvents.
The use of grease-based polishing compounds, such as are commonly used throughout the field, makes no sense at all. A major reason for cleaning parts in the first place is to get organic contaminants such as pump oils and greases, fingerprints,etc., off them. Why then do people go about doing so by coating the parts with a greasy polishing compound? This is like taking a bath in a mud puddle! Not only does the greasy polishing compound thoroughly coat the parts, but it gets embedded in every crack and crevice in them. To remove it you then have to wash the parts repeatedly in organic solvents which are expensive to purchase and also to dispose of.
Methods for cleaning vacuum parts without using greas-based polishing compounds are discussed in some detail in Sect. 2.10.4c (p. 69) of my book "Vacuum Methods in Electron Microscopy" (for info see: http://www.2spi.com/catalog/books/book48.html and http://www.bookshop.co.uk/portland/).
Since I have outlined these methods generally on this list server on a couple of previous occasions, I will not do so again. However, I would like to mention a very useful cleaning agent that is not discussed there: Tilex Soap Scum Remover. This is a combination of diethylene glycol butyl ether (a very effective solvent) and powerful detergents. It is usually possible to clean most parts simply by scrubbing them with TSSR (treating ultrasonically if desired), rinsing with hot running tap water (again treating ultrasonically if desired), rinsing with reagent grade isopropyl alcohol, and drying with a gas blaster. Check my book for more details and other useful cleaning materials.
A word of caution: the pole pieces of magnetic lenses are usually made of a soft iron alloy which corrodes rather easily, and so should not be cleaned with water; in fact, they should not be cleaned at all unless it is absolutely necessary to do so.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Wil Bigelow writes ... } } There has been a lot of discussion recently about } substituting one DP oil for another. Here is a summary of } the properties of some of the common brands of oil ...
First of all, thanx for the summary. However, I wish you would have included one other factor ... that being, whether the oil is silicone or hydrocarbon based. My impression is EM manufacturers do not suggest silicone based oils because the contamination is difficult to remove and is non-conductive. It ^has^ been a long time since having researched this issue ... I'll expect to be corrected/enlightened if I'm wrong ... it may even be possible you've listed only hydrocarbon based DP oils ...
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
at National University of Ireland, Galway ------------------------------------------------------- Papers, Posters, Workshop, Trade Exhibition -------------------------------------------------------
Guest Speakers
Professor Brigid Heywood, Keele University, England
"Bio-inspired strategies for materials fabrication"
Professor Tom Fleming, Southhampton, England
"Biological application of confocal microscopy to the study of early mammalian development"
Student prizes for papers and posters in Materials sciences and Biological sciences
------------------------------------------------------ Further details from Professor Jean Folan-Curran, Department of Anatomy, National University of Ireland, Galway. Tel: (+353) 091-750305 Fax: (+353) 091-750520 mailto:j.folan-curran-at-ucg.ie
Rather than make my way over to the library, I thought I'd poll the expertise of the group. How could I tell if some intracellular blobs are primarily lipid, primarily protein, or both/neither? Not a real analysis, but just in general. You know the things - "apears to be a lipid-like structure". This would be in plant material embedded in resin (Spurr or LX-112), and the spores are too small to see well with LM, so it rules out doing PAS at that level.
Any suggestions gratefully accepted!
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I have been freezing copepods (small planktonic crustaceans) by plunging into freezing propane, and then cryosubstituting with 2% OsO4 in anhydrous acetone (switching to methanol next week). The holder I have jury-rigged I think is not an adequate sink to slow the warming, and so I would like to have our shop mill out an aluminum block for me. I know there have been at least a couple of papers giving the dimensions of such a block and it's thermal properties, but I can't lay my hands on them at the moment. I would appreciate hearing from anyone who can suggest a reasonable start.
Also, I would like to hear from anyone with experience with ultrarapid freezing of marine (wet and salty) organisms, both plant and animal. Any particular problems or successes?
Thanks, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I am looking for prepared TEM grids for a variety of different samples (vertebrate tissues, invertebrate, plants, bacteria, viruses) that anyone may be willing to part with for the cost of shipping. I currently operate a functional Zeiss 9A TEM which I demonstrate to our undergrad. students but we are still several months away from having the ability to prepare our own grids. Thanks in advance for your help.
Mike
Michael A. Palladino, Ph.D. Instructor of Biology Brookdale Community College Lincroft, NJ 07738 Voice: (732) 224-2871 E-mail: mpalladino-at-brookdale.cc.nj.us
I am looking for prepared TEM grids for a variety of different samples (vertebrate tissues, invertebrate, plants, bacteria, viruses) that anyone may be willing to part with for the cost of shipping. I currently operate a functional Zeiss 9A TEM which I demonstrate to our undergrad. students but we are still several months away from having the ability to prepare our own grids. Thanks in advance for your help.
Mike
Michael A. Palladino, Ph.D. Instructor of Biology Brookdale Community College Lincroft, NJ 07738 Voice: (732) 224-2871 E-mail: mpalladino-at-brookdale.cc.nj.us
Hi to all fellow microscopists, I have appealed via a colleague before about this problem and thought I had solved it, but to no avail. We are looking for anyone selling parts from an Hitachi H-600 TEM. We need to replace the gun and cable and as yet have not had a promising response. The agents will not even give a price, as I believe this part is not stocked anymore.HELP from anyone who has information that I can use Bruce Dando
} Hello, all } } Rather than make my way over to the library, I thought I'd poll the } expertise of the group. How could I tell if some intracellular blobs are primarily lipid, primarily protein, or both/neither? Not a real analysis, but just in general. You know the things - "apears to be a lipid-like structure". This would be in plant material embedded
Tina:
In animal material lipid droplets are not surrounded by a membrane while protein and carbohydrate granules are. See the chapter on Adipose tissue in almost any edition of "A Textbook of Histology" by Bloom and Fawcett for an micrograph of this (taken by one of my mentors, Eunice Wood).
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Some time ago there was a thread regarding using a gold coin as a replacement for the manufacturer's gold target on sputtering units.
If there is anyone familiar with the topic, I could use a little help. Standard gold coins are 99.9% gold, can I assume this is pure enough? Do the coins need to be smoothed out? And can they be soldered in, or is brazing prefered?
Any additional hints would be greatly appreciated.
Daraporn Arayasantiparb wrote: =============================================== Would anyone be able to recommend us on what diamond knife angle would be a better choice for cutting polymer (polymer-metal/glass fiber composite)? 45 degree or 35 degree or doesn't matter?
I was told by Diatome that 35 degree knife would be a better choice to cut polymers, i.e., it reduces the curling problem. But we are currently using the 45 degree knife. Would it be wise to resharpen the one we have or buy a new one that is 35 degree? ================================================ We have cut samples of this type for more years than I would care to admit. You did not mention the metal but let me assume it is something not too hard like aluminum or copper. The hardness of the metal and its size is the single most important factor in assessing possibilities of sectioning this particular kind of system.
It is a fact that the smaller knife angle does give all kinds of advantages, another one being that it will also be easier and faster to get "acceptable sections" however you want to measure "acceptable".
But what is left unsaid is the accelerated wear rate of the knife edge. It has at least been our own experience that a 35 deg. relative to 45 deg. will "wear out" far faster, perhaps a factor of ten faster. On the other hand, if you are experienced with the microtomy of these kinds of samples, or once you become experienced, you can overcome this issue to the point that you can (usually) obtain samples that are quite acceptable fairly quickly with the 45 deg. edge. But there will always be some samples at the margin where this is not going to be the case. There will be some samples that just can not be cut at 45 deg. but which can be cut at 35 deg. Now we love these kinds of samples since they can consume a lot of diamond knives in a short period of time.
We also find that this kind of sample is more easily cut at RT than cryo. Those at the "margin" can't be cut cryo at all.
We use in our own laboratory our own SPI Materials Science Diamond Knife, 45 deg. edge for this type of sample. Depending on the hardness of the metal particles and the adhesion of the filler to matrix, there can be some pull out. But the sections are basically usable and the knife does not wear out in an afternoon. The hardness of the metal will determine how fast there is developed a pattern of "knife marks" at the edge. The next most important factor is the diameter of the glass fibers.
So the real answer to your question is a "we don't know". But if you have the 45 deg. knife anyhow, it would seem that you should not assume the worst case scenario, have it sharpened and see what you can do with it.
Disclaimer: SPI has offered a special "materials science diamond knife" with standard 45 deg. angle for some years so we have a vested interest in seeing more use of diamond knives for this kind of sample. And, yes, we are a competitor of Diatome.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Dear Listers, Wil Bigelow recently posted a message on the properties of various DP oils. We are using a fluorinated oil called Krytox 1525, which was not one of the oils discussed. Wil, do you (or anyone else) know the MM, VP, BT & HV of this oil? TIA. Yours, Bill Tivol
Early versions of the JEOL 100CX TEM use 16.2volt and 8.1volt mercury batteries as reference voltage source. There is also a 29.7volt mercury battery in the objective lens circuit.
Now that mercury batteries are no longer available, can anyone tell me if there are any suitable substitutes available, preferably in the UK?
Regards,
Bob Phillips
MicroServiS
11 Grafton Close, St. Ives, Huntingdon, http://dspace.dial.pipex.com/microservis/ Cambs., PE17 6DL
} } Some time ago there was a thread regarding using a gold coin as a } replacement for the manufacturer's gold target on sputtering units. } } If there is anyone familiar with the topic, I could use a little help. } Standard gold coins are 99.9% gold, can I assume this is pure enough? Do } the coins need to be smoothed out? And can they be soldered in, or is } brazing prefered? }
Any half way competent high street jeweller can hammer out a pure solid gold target for your sputter coater.
Ours are 0.5 mm thick, 70 mm diameter.
They can start with a gold coin, or will make it out of any weight of pure gold you specify.
Once it is worn through, you take the remnant back to the jeweller and ask him to make it up back to the original weight.
It is never wasted........
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
I've been asked to put out a call for help in sourcing a cheap (as possible) X100 air spaced objective with a minimal working distance of 0.4mm but preferably much longer (ideal 4mm) and a high NA greater than 0.65 (double would be good).
This seems a lot to ask but i'm sure someone out there can detail a best fit to these requirements.
Many thanks
Dr. Mark Osborne
Department of Chemistry Uni of Cambridge Lensfield Rd UK CB2 1EW
All you need to make a piece of metal the target of your coater is to fix=
it in place such that it makes a good contact with the power source. =
Provided the base unit upon which it is mounted is aluminium you will not=
have any sputtering from the base. Hammering the coin flat will give you= a bigger source area of course.
Many targets are clipped in place, a poor contact may be improved with th= e use of conducting paste the same as you would use to fix a specimen to to=
stub.
The material should be more than adequate for Tungsten SEM applications.
For the first sputter do not use a specimen but let the current run so as=
to clean the target.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
I have had to repair HT cables in many countries of the world where it wa= s rare for a new HT cable to be on hand. It is not the problem that you m= ay be expecting.
There are many other industries that use HT cables, main power lines are = a good example. Look in your local Yellow pages for organisations in the fields of - power cables, x-ray generators, capacitor manufacturers, hig= h voltage transformer manufacturers etc.
They will be able to use your Hitachi cable ends and simply replace and r= e seal the cable.
Best of luck
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Our electronics workshop on campus designed two circuits which reduce the dependance of the JEOL 100CX on expensive batteries. Please let me know if you would like to receive info about the circuits, which have served our 100CX reliably for more than 5 years.
Tony
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Private Bag X01, Scottsville 3209, KwaZulu-Natal, South Africa. Tel +27 (0)331 260 5155, Home +27 (0)331 962676 Fax +27 (0)331 260 5776 email: bruton-at-emu.unp.ac.za
Depending on the current and stability requirements... You might want to check out replacing the batteries with active solid state devices. Although more stable references are available, the LM317 adjustable voltage regulator (for less than $2.00) may do the job. The output voltage is set using a two resistor voltage divider and an internal reference. Be sure to use stable resistors - wirewound or metal film.
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello fellow subscribers,
Early versions of the JEOL 100CX TEM use 16.2volt and 8.1volt mercury batteries as reference voltage source. There is also a 29.7volt mercury battery in the objective lens circuit.
Now that mercury batteries are no longer available, can anyone tell me if there are any suitable substitutes available, preferably in the UK?
Regards,
Bob Phillips
MicroServiS
11 Grafton Close, St. Ives, Huntingdon, http://dspace.dial.pipex.com/microservis/ Cambs., PE17 6DL
id AA17924 for Microscopy-at-Sparc5.Microscopy.Com; Fri, 5 Jun 98 05:02:25 MST Received: from msgphx3.sps.mot.com by mogate.sps.mot.com (4.1/SMI-4.1/Email-2.0) id AA16550 for Microscopy-at-Sparc5.Microscopy.Com; Fri, 5 Jun 98 05:02:24 MST Received: from email.sps.mot.com ([221.105.10.56]) by msgphx3.sps.mot.com (Netscape Messaging Server 3.01) with ESMTP id AAA29662; Fri, 5 Jun 1998 05:02:22 -0700 Message-Id: {3577DDD4.7D454685-at-email.sps.mot.com}
Bob,
I had the same problem with my cameras, the substitution that we made was a silver/air or zinc/air cell. It's about 5:00 AM here and I don't recall clearly which metal system is used. The cameras are elsewhere so I can't check. These cells have the same voltage output, my cameras work fine with this substitution. The life of these cells isn't quite as good, one opens an air seal on the battery to start the cell. Hope this somewhat fuzzy information helps!!
Bob Phillips wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello fellow subscribers, } } Early versions of the JEOL 100CX TEM use 16.2volt and 8.1volt mercury } batteries as reference voltage source. There is also a 29.7volt mercury } battery in the objective lens circuit. } } Now that mercury batteries are no longer available, can anyone tell me if } there are any suitable substitutes available, preferably in the UK? } } Regards, } } Bob Phillips } } MicroServiS } } 11 Grafton Close, } St. Ives, } Huntingdon, http://dspace.dial.pipex.com/microservis/ } Cambs., } PE17 6DL
Hello all, Since the source of gold is up, I'll toss in my 2 cents worth. Years ago we too thought of the gold coin. Looking into it we found that most gold coins were not near 99.9%. It's just too soft & of course folks wanted more than the gold value for their coins. The specialty suppliers can be quite expensive (say a foctor of 3-4). The deal of the day was to purchase "gold shot". This is the stock from which jewelers make their alloys (10kt, 18kt, etc.) The purity was claimed to be 99.99% & it cost us the current exchange rate +~$10US.
Position announcement: Member, Clinical Studies Staff
Medjet Inc., a publicly held research and development firm specializing in the design and development of ophthalmic surgical devices, is now interviewing for a position available within its Department of Clinical Studies. Preferred candidates will have completed a bachelors degree in either a biological science or biomedical engineering. Candidates will be expected to have prior training and/or experience with all phases of light, scanning, and transmission electron microscopy, photography, and animal handling. Prior experience with medical device design and manufacturing is a plus. The individual will assist in the coordination, execution, and documentation of research activities including clinical trials involving animal and human subjects.
Headquarters are located in Edison, NJ. Candidates will be considered until the position has been filled. Selected candidates will be expected to begin employment during early July and be flexible considering the potential for prolonged travel stays. Salary range is between $27,500-$32,500 per year in addition to a complete benefits package. Interested applicants should forward a resume with cover letter to the attention of Daniel Caruso.
Medjet Inc. Suite 301 1090 King George Post Road Edison, NJ 08837
As a result of my recent comments over this list server, I have received several requests for additional information about diffusion pump fluids. Here are the answers to those inquiries:
The chemical compositions of the oils I discussed previously are: Octoil-S: di(2-ethylhexyl)-sebacate DC-704: tetraphenyl-tetramethyl-trisiloxane DC-705: pentaphenyl-trimethyl-trisiloxane Santovac-5: mixture of five-ring polyphenylethers with MM = 454 Alcatel 220: eicosyl naphthalene
The common Krytox diffusion pump fluids are high molecular weight perfluorinated-alkylpolyether (perfluoroalkyl ether) fluids, manufactured by the DuPont Company, with approximately the following properties (in the same units given previously):
I would repeat, all this information, and many other goodies, can be found in my book "Vacuum Methods in Electron Microscopy" (for info about it see: http://www.2spi.com/catalog/books/book48.html and http://www.bookshop.co.uk/portland/).
I might also comment that another source has indicated that Lion-S DP oil is eicosyl naphthalene, similar to the Alcatel-200 fluid, rather than di-(2-ethylhexyl)-sebacate, like Octoil-S.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Earlier I received many helpful comments regarding the different types of SEM's. My question now is what experiences do users of FE-SEM's have with cold cathode field emitters vs. thermal field or Schottky field emitters. Please include comments on resolution, vacuum problems and requirements, and interference from building and/or room environmental conditions. Samples will include both textiles and biologicals. Thank you in advance.
Bruce F. Ingber Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124
} Earlier I received many helpful comments regarding the different types of } SEM's. My question now is what experiences do users of FE-SEM's have with } cold cathode field emitters vs. thermal field or Schottky field emitters. } Please include comments on resolution, vacuum problems and requirements, } and interference from building and/or room environmental conditions. } Samples will include both textiles and biologicals. Thank you in advance. } } Bruce F. Ingber } Biologist- Electron Microscopy } USDA-ARS, SRRC } 1100 Robert E. Lee Blvd. } New Orleans, LA 70124 } } (504) 286-4270; fax (504) 286-4419 } bingber-at-nola.srrc.usda.gov }
Thank you for the replies on the microwave techniques course and fluorescent cell membrane marker information requests. For the people who are interested, on florescent cell membrane labeling, DiI DiO and NBD or BODIPY-labeled phospholipids were suggested. It was strongly recommended to refer to Molecular Probes catalogue, which indeed is a very good source of information. Thank you for help again, Lilith --------------------------
What is a good procedure for rotationally aligning the upper Wollaston prism on a Zeiss microscope's DIC optical system? (By "upper" is meant the one closest to the light-source. This element is a part of the condensor assembly).
Many thanks for a workable answer.
Robley Williams (robley.williams-at-vanderbilt.edu)
I am trying to use a SIS slow scan CCD (Biocam) with the Philips CM spotscan module to image viruses at low dose. I wonder if anyone has any experience with this combination of microscope, camera and spot-scanning? Could you give me advice or tips as to how to do it?
Many thanks,
Dr Timothy F. Booth Canadian Food Inspection Agency National Centre For Foreign Animal Disease Suite T2300 1015 Arlington St. Winnipeg Manitoba R3E 3M4 CANADA http://www.hc-sc.gc.ca/main/lcdc/web/bmb/fedlab_e.html#toc email tbooth-at-em.agr.ca Tel 204 789 2022 Fax 204 789 2038
A fellow called Bob was asking about innovative solutions to expensive battery replacements in the JEOL 100CX. We posted a solution to this on the server some time ago but couldn't have seen it. His mail will not respond to me -
Bob - if you are out there then get in touch with me directly and I will tell you how we tackled the same problem with great success.
Tony
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Private Bag X01, Scottsville 3209, KwaZulu-Natal, South Africa. Tel +27 (0)331 260 5155, Home +27 (0)331 962676 Fax +27 (0)331 260 5776 email: bruton-at-emu.unp.ac.za
Your message was sent a few weeks ago and it took me a while to find the = references that I wanted to send to you. You can look at the strain in = a heterostructure with convergent beam electron diffraction analysis. = However, several years ago, Hamish Frazier and company published some = work in the MSA proceedings regarding the errors of doing this with CBED = on XTEM because of surface relaxation in the sample. You should look it = up, it was about 5-6 years ago. Doug Perovic et al. also wrote two = excellent papers on this topic. In these papers, they modeled image of = the layers and included the contribution of surface relaxation effects = to the contrast in the image. I'm sure that there are other papers on = this topic, but these are what I am readily aware of and they are very = good. Here are the references:=20
"On the electron diffraction contrast of coherently strained = semiconductor layers", D. D. Perovic et al., Phil. Mag. A,, 64(1), pp. = 1-28, 1991.
"On the electron microscope contrast of doped semiconductor layers," D. = D. Perovic et al., Phil. Mag. A,, 63(4), pp. 757-784, 1991.
} microscopy-at-MSA.microscopy.com } ---------- } From: feng-at-iris.lamel.bo.cnr.it } To: microscopy-at-sparc5.microscopy.com } Subject: relaxation in TEM specimen } Date: Thursday, May 21, 1998 5:44AM } } =20 } ------------------------------------------------------------------------=
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Dear Sir: } I am now doing some strain analysis in the herostructure ( one } called LOCOS). At the moment, I only have result based on bulk } materials. Do } you know some good way to take into account the relaxation in this kind } of } semiconductor? } } Thanks inadvance. } } Sincerely, } } Feng Wu } =20
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 = HTML//EN"} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} Your message was sent a few weeks ago and it took me a while to = find the=20 references that I wanted to send to you. You can look at the = strain in a=20 heterostructure with convergent beam electron diffraction = analysis. =20 However, several years ago, Hamish Frazier and company published some = work in=20 the MSA proceedings regarding the errors of doing this with CBED on XTEM = because=20 of surface relaxation in the sample. You should look it up, it was about = 5-6=20 years ago. Doug Perovic et al. also wrote two excellent papers on = this=20 topic. In these papers, they modeled image of the layers and = included the=20 contribution of surface relaxation effects to the contrast in the = image. =20 I'm sure that there are other papers on this topic, but these are what I = am=20 readily aware of and they are very good. Here are the=20 references: {/DIV} {DIV} {/DIV} {DIV} "On the electron diffraction contrast of coherently strained=20 semiconductor layers", D. D. Perovic et al., {U} Phil. Mag. A, {/U} ,=20 {U} 64 {/U} (1), pp. 1-28, 1991. {/DIV} {DIV} {/DIV} {DIV} "On the electron microscope contrast of doped semiconductor=20 layers," D. D. Perovic et al., {U} Phil. Mag. A, {/U} ,=20 {U} 63 {/U} (4), pp. 757-784, 1991. {/DIV} {DIV} {/DIV} {DIV} {/DIV} {DIV} > {A=20 href=3D"mailto:microscopy-at-MSA.microscopy.com"} microscopy-at-MSA.microscopy.c= om {/A} {BR} >=20 ---------- {BR} >From: {A=20 href=3D"mailto:feng-at-iris.lamel.bo.cnr.it"} feng-at-iris.lamel.bo.cnr.it {/A} {B= R} >To:=20 {A=20 href=3D"mailto:microscopy-at-sparc5.microscopy.com"} microscopy-at-sparc5.micros= copy.com {/A} {BR} >Subject:=20 relaxation in TEM specimen {BR} >Date: Thursday, May 21, 1998=20 5:44AM {BR} > {BR} >=20 {BR} >-----------------------------------------------------------------= ------- {BR} >The=20 Microscopy ListServer -- Sponsor: The Microscopy Society of=20
Your message was sent a few weeks ago and it took me a while to find the = references that I wanted to send to you. You can look at the strain in = a heterostructure with convergent beam electron diffraction analysis. = However, several years ago, Hamish Frazier and company published some = work in the MSA proceedings regarding the errors of doing this with CBED = on XTEM because of surface relaxation in the sample. You should look it = up, it was about 5-6 years ago. Doug Perovic et al. also wrote two = excellent papers on this topic. In these papers, they modeled image of = the layers and included the contribution of surface relaxation effects = to the contrast in the image. I'm sure that there are other papers on = this topic, but these are what I am readily aware of and they are very = good. Here are the references:=20
"On the electron diffraction contrast of coherently strained = semiconductor layers", D. D. Perovic et al., Phil. Mag. A,, 64(1), pp. = 1-28, 1991.
"On the electron microscope contrast of doped semiconductor layers," D. = D. Perovic et al., Phil. Mag. A,, 63(4), pp. 757-784, 1991.
-Scott Walck
******************************************** Scott D. Walck, Ph.D. (Work Email: Walck-at-PPG.com) Karen L. Walck Adam S. Walck Brian D. Walck=20 4718 Denbigh Ct. Allison Park, PA 15101 (412) 492-8127 ********************************************=20 } microscopy-at-MSA.microscopy.com } ---------- } From: feng-at-iris.lamel.bo.cnr.it } To: microscopy-at-sparc5.microscopy.com } Subject: relaxation in TEM specimen } Date: Thursday, May 21, 1998 5:44AM } } =20 } ------------------------------------------------------------------------=
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Dear Sir: } I am now doing some strain analysis in the herostructure ( one } called LOCOS). At the moment, I only have result based on bulk } materials. Do } you know some good way to take into account the relaxation in this kind } of } semiconductor? } } Thanks inadvance. } } Sincerely, } } Feng Wu } =20
******************************************** Scott D. Walck, Ph.D. (Work Email: Walck-at-PPG.com) Karen L. Walck Adam S. Walck Brian D. Walck 4718 Denbigh Ct. Allison Park, PA 15101 (412) 492-8127 ********************************************
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {DIV} Your message was sent a few weeks ago and it took me a while to = find the=20 references that I wanted to send to you. You can look at the = strain in a=20 heterostructure with convergent beam electron diffraction = analysis. =20 However, several years ago, Hamish Frazier and company published some = work in=20 the MSA proceedings regarding the errors of doing this with CBED on XTEM = because=20 of surface relaxation in the sample. You should look it up, it was about = 5-6=20 years ago. Doug Perovic et al. also wrote two excellent papers on = this=20 topic. In these papers, they modeled image of the layers and = included the=20 contribution of surface relaxation effects to the contrast in the = image. =20 I'm sure that there are other papers on this topic, but these are what I = am=20 readily aware of and they are very good. Here are the=20 references: {/DIV} {DIV} {/DIV} {DIV} "On the electron diffraction contrast of coherently strained=20 semiconductor layers", D. D. Perovic et al., {U} Phil. Mag. A, {/U} ,=20 {U} 64 {/U} (1), pp. 1-28, 1991. {/DIV} {DIV} {/DIV} {DIV} "On the electron microscope contrast of doped semiconductor=20 layers," D. D. Perovic et al., {U} Phil. Mag. A, {/U} , = {U} 63 {/U} (4),=20 pp. 757-784, 1991. {/DIV} {DIV} {/DIV} {DIV} -Scott Walck {/DIV} {DIV} {/DIV} {DIV} {/DIV} {DIV} {/DIV} {DIV} {DIV} {/DIV} {DIV} {FONT color=3D#000000=20 size=3D2} ******************************************** {BR} Scott D. Walck, =
Ph.D. (Work Email: {A=20 href=3D"mailto:Walck-at-PPG.com"} Walck-at-PPG.com {/A} ) {BR} Karen L. = Walck {BR} Adam S.=20 Walck {BR} Brian D. Walck {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} 4718 Denbigh Ct. {BR} Allison Park, = PA =20 15101 {BR} (412)=20 492-8127 {BR} ******************************************** {/FONT} {/DI= V} {/DIV} {DIV} > {A=20 href=3D"mailto:microscopy-at-MSA.microscopy.com"} microscopy-at-MSA.microscopy.c= om {/A} {BR} >=20 ---------- {BR} >From: {A=20 href=3D"mailto:feng-at-iris.lamel.bo.cnr.it"} feng-at-iris.lamel.bo.cnr.it {/A} {B= R} >To:=20 {A=20 href=3D"mailto:microscopy-at-sparc5.microscopy.com"} microscopy-at-sparc5.micros= copy.com {/A} {BR} >Subject:=20 relaxation in TEM specimen {BR} >Date: Thursday, May 21, 1998=20 5:44AM {BR} > {BR} >=20 {BR} >-----------------------------------------------------------------= ------- {BR} >The=20 Microscopy ListServer -- Sponsor: The Microscopy Society of=20
Ph.D. (Work Email: {A=20 href=3D"mailto:Walck-at-PPG.com"} Walck-at-PPG.com {/A} ) {BR} Karen L. = Walck {BR} Adam S.=20 Walck {BR} Brian D. Walck {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} 4718 Denbigh Ct. {BR} Allison Park, = PA =20 15101 {BR} (412)=20 492-8127 {BR} ******************************************** {/FONT} {/DI= V} {/BODY} {/HTML}
Journal : Micron Volume/issue : 29/2-3, Year : 1998, ISSN : 0968-4328 Cover Date : April-June 1998
The combination of high-accelerator epoxy resin and antigen retrieval to obtain more intense immunolabeling on epoxy sections than on LR-White sections for large proteins S-H Brorson
Low energy loss electron microscopy of chromosomes MMG Barfels, X Jiang, YM Heng, AL Arsenault, FP Ottensmeyer pp. 105-111 Experimental model to study sedimentary kidney stones F Grases, A Llobera
Pseudo-aberration free focus condition for atomic resolution electron microscope images H Hashimoto, H Endoh, M Hashimoto, ZP Luo, MF Song
The life cycle of human immunodeficiency virus type 1 T Goto, M Nakai, K Ikuta
Transmitted color and interference fringes for tem sample preparation of silicon JP Mccaffrey
Cryo-negative staining M Adrian, J Dubochet, SD Fuller, JR Harris
Symmetry in the 2.25 MDa homomultimeric phosphoenolpyruvate synthase from Staphylothermus marinus. Analyses of negatively stained preparations G Harauz
Vesicle to micelle structural transitions involved in the interaction of dodecylbetaine with liposomes: transmission electron microscopy and light scattering studies A De La Maza, L Coderch, O Lopez, JL Parra
TEM, STM and AFM as tools to study clusters and colloids GL Hornyak, S Peschel, T Sawitowski, G Schmid
High-spatial resolution compositionally-sensitive imaging of metallic particles using plasmon energy-loss electrons in TEM EM Hunt, ZL Wang, ND Evans, JM Hampikian
Atomic force microscopy imaging of the growth features on the surface of rutile F Czerwinski, JA Szpunar
The ultrastructure of yeast: Cell wall structure and formation M Osumi
The effect of lithium treatment on collagenous tissues: an electron microscope study M Tzaphlidou, E Kounadi
Colour atlas of basic histology JR Harris
The Golgi apparatus, edited by EG Berger and J Roth JR Harris
Dear helpful readers!
I was told there are five different kinds of target available for sputtering unit and they are Gold, Gold-Palladium, Platium, Nikel and Silver
Could anyone please explain for me about the use of these targets.
What are the advantage and disadvantage on each of them? What are the effects on the quality of the image when we use the "wrong" target material? Which target material is most suitable for which kind of sample?
Any information about target material would be greatly appreciated
Thanks
Khanh Tran Khanh Tran Deakin University 662 Blackburn Road CLAYTON, VIC. 3168 AUSTRALIA
I was told there are five different kinds of target available for the sputtering unit and they are Gold, Gold-palladium, Platium, Nikel and silver
Could anyone please explain for me about the use of these targets.
What are the advantage and disadvantage on each of them? What are the effects on the image when we use the "wrong" target material? Which target is most suitable for which kind of sample?
Any information about target material would be greatly appreciated
Thanks
Khanh Tran Khanh Tran Deakin University 662 Blackburn Road CLAYTON, VIC. 3168 AUSTRALIA
there are even more. Carbon, chromium, tantalum etc.
Coatings are applied to make the sample conductive. The necessary conductivity depends on your SEM. Field-emission SEMs do not need much, so that we do not sputter most of our samples.
The disadvantage is of course that these coatings are like covers - hiding the underlying sample. You need a minimum thickness which is 2-3nm of Au-Pd or 8-12nm of chromium for most of our samples. The coating is often not smooth but shows islands. The coating smoothness depends on the sputter conditions (vacuum, sample cooling, sample rotation) and on the material. Alloys are often better than pure metals because they do not crystallize so easily. We see Au-Pd as a very good compromise (having more structure than Cr but needs much less thickness).
Khanh Tran wrote:
} I was told there are five different kinds of target available for the } sputtering unit and they are Gold, Gold-palladium, Platium, Nikel and silver } } Could anyone please explain for me about the use of these targets. } } What are the advantage and disadvantage on each of them? } What are the effects on the image when we use the "wrong" target material? } Which target is most suitable for which kind of sample?
I am interested in any information as to the use of latex microspheres for low mag. calibration. I am particularly interested in the shelf-life and stability of diluted suspensions (being able to use the diluted suspension more than one time).
First I will apologize right away to those who feel that this is off topic for the mailing list. I feel it is important to small vendors and those users who feel that it is important for vendor participation at meetings.
I known that several vendors lurk this list so I want to raise a concern about some information that has been brought to my attention. The following is included with the general information for vendors that are exhibiting at MAS/MSA '98:
"NOTE: All displays must be in the process of being installed by 6:00PM, Saturday, July 11. After that time, any unattended booths with crated displays will be set up at the discretion of M&M '98 Show Management, and all expenses will be charged to the exhibitor. Any empty booths will be reassigned by Show Management at 4:00PM Saturday, July 11 (without prior written notification). All displays must be fully installed by 11:00PM Sunday, July 12."
As a small vendor, this is totaly unacceptable and will not be tolerated. Many small vendors routinely arrive late on Saturday and setup on Sunday. To place such restrictions is not acceptable. I'm paying good money and time to support the exhibition and will not be dictated to as to when we arrive and setup. Even more so when that information arrives after we have already made travel plans. These types of stupid restrictions are exactly why vendor participation is decreasing. I realize that the Show Management would like the crates removed as soon as possible so they can install carpet and such. But to say that empty booths will be reassigned by 4:00PM on Saturday. This is going much too far.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
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Dear Friends,
I am currently looking at the growth/ interface studies of oxygen on silver. Does anyone know of work/ publications in this area.
regards
Parmjit
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begin: vcard fn: Parmjit S. Flora n: Flora;Parmjit S. org: University of Alberta adr: University of Alberta, Department of Physics, Faculty of Science;;412 Avadh Bhatia Physics Laboratory;Edmonton;Alberta;T6G 2J1;Canada email;internet: pflora-at-laser.phys.ualberta.ca title: Dr. tel;work: 403-492-4123 tel;fax: 403-492-0714 tel;home: 403-482-5840 x-mozilla-cpt: ;-1 x-mozilla-html: FALSE version: 2.1 end: vcard
I am interested in any information as to the use of latex microspheres for low mag. calibration. I am particularly interested in the shelf-life and stability of diluted suspensions (being able to use the diluted suspension more than one time).
I came across this article in hardcopy form while waiting for a recent dentist appointment. Its a rather longish look at ethical concerns regarding nature photography (e.g., National Geographic, and others). It seems microscopists aren't the only group grappling with where the fine line is between cleaning up an image for publication and manipulating the data.
"Photography in the Age of Falsification" by: Kenneth Brower, Atlantic Monthly, May 1998
http://www.theatlantic.com/issues/98may/photo.htm
Yours, Doug ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
Hello! I am working on a Murdock Grant this summer and am having a difficult time locating information on TEM specimen preparation for frog abdominal skin. A colleague and I are studying acid toxicity of frog epithelial tissue and need electron micrographs to observe damage done to the tight junctions due to increased pH. I have a few books on order, but am worried that they won't have the information that I need. I also need to know how to adjust my buffer systems for we are studying the ion permeability of the skin at a number of acidic pH's. I am using a glutaraldehyde/osmium tetraxide fixative and a Ringer's solution for a buffer. If you could provide me with any info, references, tips, etc. I would be very appreciative.
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Dear Friends,
I am currently looking at the growth/ interface studies of oxygen on silver. Does anyone know of work/ publications in this area.
regards
Parmjit
--------------EB0A8AC9A2353FBEE181809A Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Parmjit S. Flora Content-Disposition: attachment; filename="vcard.vcf"
begin: vcard fn: Parmjit S. Flora n: Flora;Parmjit S. org: University of Alberta adr: University of Alberta, Department of Physics, Faculty of Science;;412 Avadh Bhatia Physics Laboratory;Edmonton;Alberta;T6G 2J1;Canada email;internet: pflora-at-laser.phys.ualberta.ca title: Dr. tel;work: 403-492-4123 tel;fax: 403-492-0714 tel;home: 403-482-5840 x-mozilla-cpt: ;-1 x-mozilla-html: FALSE version: 2.1 end: vcard
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Parmjit S. Flora wrote:
} Dear Friends, } } I am currently looking at the growth/ interface studies of oxygen on } silver. Does anyone know of work/ publications in this area. } } regards } } Parmjit } } ------------------------------------------------------------------------ } } Parmjit S. Flora {pflora-at-laser.phys.ualberta.ca} } Dr. } University of Alberta } } Parmjit S. Flora } Dr. {pflora-at-laser.phys.ualberta.ca} } University of Alberta } University of Alberta, Department of Physics, Faculty of Science Work: 403-492-4123 } 412 Avadh Bhatia Physics Laboratory Fax: 403-492-0714 } Edmonton Home: 403-482-5840 } Alberta Netscape Conference Address } T6G 2J1 } Canada } Additional Information: } Last Name Flora } First Name Parmjit S. } Version 2.1
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begin: vcard fn: Parmjit S. Flora n: Flora;Parmjit S. org: University of Alberta adr: University of Alberta, Department of Physics, Faculty of Science;;412 Avadh Bhatia Physics Laboratory;Edmonton;Alberta;T6G 2J1;Canada email;internet: pflora-at-laser.phys.ualberta.ca title: Dr. tel;work: 403-492-4123 tel;fax: 403-492-0714 tel;home: 403-482-5840 x-mozilla-cpt: ;-1 x-mozilla-html: FALSE version: 2.1 end: vcard
Does anyone know of a way to visualize the tempering on a piece of window = safety glass? I've tried cross sectioning followed by etching with 10% = HFwith no indication of any differences through the thickness of the = glass.
The only thing I've noticed so far is a difference in the fracture = surface pattern near the edge of the glass (about 1 mm on each side). = Near the edge a ridge pattern runs perpendicular to the glass surface = while in the center the fractured surface is mostly smooth with some = ridging running roughly parallel to the surface in the middle. (Total = thickness of the glass is about 4 mm.)
Am I seeing tempering in the 1 mm of perpendicular ridges? Does anybody = have a ball park number for how thick a surface layer is affected by the = tempering?
Does anyone know of a way to visualize the tempering on a piece of window = safety glass? I've tried cross sectioning followed by etching with 10% = HFwith no indication of any differences through the thickness of the = glass.
The only thing I've noticed so far is a difference in the fracture = surface pattern near the edge of the glass (about 1 mm on each side). = Near the edge a ridge pattern runs perpendicular to the glass surface = while in the center the fractured surface is mostly smooth with some = ridging running roughly parallel to the surface in the middle. (Total = thickness of the glass is about 4 mm.)
Am I seeing tempering in the 1 mm of perpendicular ridges? Does anybody = have a ball park number for how thick a surface layer is affected by the = tempering?
Are there any advantages to using cacodylate buffers in fixatives as opposed to phosphate buffers? Are there any disadvantages in addition to the toxicity of cacodylate buffers?Specifically, I am working with human corneas. Thanks for your replies.
drose-at-wlgore.com wrote the following: =================================================== I am interested in any information as to the use of latex microspheres for low mag. calibration. I am particularly interested in the shelf-life and stability of diluted suspensions (being able to use the diluted suspension more than one time). ==================================================== You did not mention whether for SEM or TEM but I will assume it is SEM. Your question is particularly hard to answer since the surfactant systems vary so widely but generally speaking, the big concern is if there has been any kind of coagulation of the latex. If you shake up the vial, and don't "see" anything floating, then it is likely that it is OK. We have had diluted material at times years old which still seemed to be OK. We have diluted other latex and it seems to destabilize in just a few days.
But as a word of caution, the lower the magnification, the larger the latex sphere and the larger the sphere, the larger the expected standard deviation and also, large spheres will become unstable faster than will small ones.
Unless you have some really out-of-the-ordinary need, you might be better off using one of the calibrated SEM mounts made by electron beam lithography methods, etched into silicon. You can deposit small samples on that kind of sample which is the most useful in fact at "low magnifications" and you also have a form of built in calibration "yardstick". It can also be recleaned and reused.
You can see one such example on our website at URL http://www.2spi.com/catalog/standards/stndcal1.html
We are not the only supplier for this kind of product, most of the main suppliers of EM consumables offer their own versions which at least to the first approximation, will do the same thing.
Disclaimer: SPI Supplies offers both calibrated microspheres and a calibrated SEM mount, however, we do believe that for low magnification SEM work, the calibrated SEM mount is a better choice.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
1. Cacodylate stocks will keep longer, not allowing fungi to grow.=20 2. Cacodylate has aditional fixing activity of its own 3. Phosphate may originate some precipitates in some materials.
Disadvantages: Will precipitate with uranyl acetate (as well as phosphate). So, replace = the buffer before using uranyl acetate "in block" staining.
Dr. A.P. Alves de Matos mtlopes-at-fc.ul.pt
----- Mensagem original ----- De: Dr. Gernot Richter [SMTP:mdjt-clin-at-monmouth.com] Enviada em: Ter=E7a-feira, 9 de Junho de 1998 2:47 Para: Microscopy-at-sparc5.microscopy.com Assunto: Cacodylate buffers
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Hello everyone,
Are there any advantages to using cacodylate buffers in fixatives as opposed to phosphate buffers? Are there any disadvantages in addition to the toxicity of cacodylate buffers?Specifically, I am working with human corneas. Thanks for your replies.
1. If you add calcium to your fixative/buffer, cacodylate would avoid precipitation of insoluble calcium phosphate.
2. Micro organisms do not grow in cacodylate buffer as they do in phosphate buffer. So you must prepare fresh phosphate more frequently...you may also store phosphate buffer in the freezer and thaw as needed.
On Mon, 8 Jun 1998, Dr. Gernot Richter wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello everyone, } } Are there any advantages to using cacodylate buffers in fixatives as } opposed to phosphate buffers? Are there any disadvantages in addition to } the toxicity of cacodylate buffers?Specifically, I am working with human } corneas. Thanks for your replies. } } Dan Caruso } Medjet Inc. } } }
************************************************************************** Delilah F. Wood Tel: 510-559-5653 USDA - ARS - WRRC Fax: 510-559-5777 800 Buchanan St. Email: wood-at-pw.usda.gov Albany, CA 94710 **************************************************************************
First, let me apologize to the vast majority of you, who are not affected by exhibitor procedures for Microscopy and Microanalysis '98, for bothering you with this, but Scott Davilla raised some concerns yesterday in this forum that need a response. Hopefully this will end this discussion.
Scott was correctly objecting to a specific section of the instructions to exhibitors concerning booth set-up times. This was inadvertently included in the M&M'98 instructions by our new Meeting Manager and does not apply to M&M'98. Any exhibitor concerned about this or any other aspect of M&M'98 is encouraged to contact the Meeting Manager via phone (708-361-6000), fax (708-361-6166) or e-mail (msa-at-tradeshownet.com). We are committed to resolving all issues in a satisfactory manner.
M&M'98, sponsored by MSA and MAS, is proud of its reputation as both the premier yearly Microscopy exhibition, and also as a meeting that cares deeply about the satisfaction of its exhibitors and scientific attendees. Speaking for MSA, we have involved the Sustaining Members Committee in all recent decision-making about the M&M meetings, and both MSA and MAS have members from exhibiting companies on their governing Councils.
Any member or attendee who has concerns about any aspect of the Meeting or membership (for MSA) can contact the Meeting Manager, the MSA Business Office, or, if that is not satisfactory, MSA Council directly. All of these groups can be accessed from the MSA WWW site, www.msa.microscopy.com.
Finally, one point raised by Scott requires correction and clarification. Building on the great success of M&M'96 in Minneapolis, the Cleveland meeting (M&M'97) broke all records for exhibit space for a recent M&M meeting. It now appears that M&M'98 in Atlanta will meet or exceed the Cleveland numbers - in fact, the Meeting Manager has just had to add 10 new booth spaces to the original plan. So we look forward to seeing you in Atlanta for what promises to be an exciting and enriching meeting.
Ernie Hall Secretary, MSA
_____________________________________________________________________________________________________ ________________________ Ernest L. Hall Manager, Microstructure and Microanalysis Program Characterization & Environmental Technology Laboratory GE Corporate Research and Development Building K-1, Room 2C12 PO Box 8 (1 Research Circle) Schenectady, NY 12301 (12309) Phone 518-387-6677; Fax -6972; Dialcomm 8-833-6677 Email: hallel-at-crd.ge.com
Does anyone know of a way to visualize the tempering on a piece of window = safety glass? I've tried cross sectioning followed by etching with 10% = HFwith no indication of any differences through the thickness of the = glass.
The only thing I've noticed so far is a difference in the fracture = surface pattern near the edge of the glass (about 1 mm on each side). = Near the edge a ridge pattern runs perpendicular to the glass surface = while in the center the fractured surface is mostly smooth with some = ridging running roughly parallel to the surface in the middle. (Total = thickness of the glass is about 4 mm.)
Am I seeing tempering in the 1 mm of perpendicular ridges? Does anybody = have a ball park number for how thick a surface layer is affected by the = tempering?
there is an excellent review of the "buffer in EM"-problem in G. Griffith's book "Fine structure in Immunocytochemistry", pp.57 Springer-Verlag, Berlin, 1993.
Hope that helps.
Heinz
} Hello everyone, } } Are there any advantages to using cacodylate buffers in fixatives as } opposed to phosphate buffers? Are there any disadvantages in addition to } the toxicity of cacodylate buffers?Specifically, I am working with human } corneas. Thanks for your replies. } } Dan Caruso } Medjet Inc.
*********************************************************************** Dr. Heinz Fehrenbach Institute of Pathology University Clinics "Carl Gustav Carus" Technical University of Dresden
I came across this article in hardcopy form while waiting for a recent dentist appointment. Its a rather longish look at ethical concerns regarding nature photography (e.g., National Geographic, and others). It seems microscopists aren't the only group grappling with where the fine line is between cleaning up an image for publication and manipulating the data. "Photography in the Age of Falsification" by: Kenneth Brower, Atlantic Monthly, May 1998 http://www.theatlantic.com/issues/98may/photo.htm
6/9/98 Reply:
For users of this listserv who haven't taken a speed reading course the above linked article covers the ethics of artistic license when applied to nature photography within this era of digital enhancement. I color microscans professionally and can easily understand the article's application to microscopy.
1-when does a digitally manipulated image become a fake. 2-how far do you go to satisfy the public's appetite for exciting images. 3-artistic expression - adding and subtracting from the image. 4-should a picture be labeled as digitally enhanced. 5-digital capabilities, is it progress or over the top.
Evidently when ethics are discussed, philosophical gray areas arise. I absolutely, without question, appreciate the importance of resolution and accuracy in the microscopist' work. However, I do believe colored scans provide entertaining education to the public. The public may not pick up on black and white photos and therefore it not available. Sometimes we don't know what color the microscopic specimens are and I am concerned about their representation. But, brown and gray is not appealing. I am also concerned about adding or subtracting parts of a subject in a microscan to gain a balanced composition. I have avoided the temptation. To lighten and darken really helps. It is now possible to do anything to an image; turn it inside out and spin it around in 3-D animation. I hope ethical discussions continue in order to provide the best scientific communication possible. Photographic digital enhancement is able satisfy public taste and how we use it is exciting new territory for communication, education, and art. All subjects that cannot be controlled but you can watch carefully.
Renee Recker Digital Design Communication Design for Biologists Web, Graphic, and Colored Microscans 16 West 16th St. NYC, NY 10011 212-675-1665 heyrenee-at-renorex.com http://www.renorex.com
Responding to the message of {Pine.SUN.3.96.980609052113.24561A-100000-at-aggie} from "Delilah F. Wood" {wood-at-aggie.pw.usda.gov} :
SNIP!
} 2. Micro organisms do not grow in cacodylate buffer as they do in } phosphate buffer.
SNIP!
One would certainly think so. However, I've noticed that 0.2 M caco. buffer, pH=7.0, after long storage times (I'm looking at a bottle of 100 ml mixed up two years ago) accumulates a bit of fluffy stuff at the bottom that is a mid-tone grey in color, but not a solid black. It kinda looks like fungal stuff, like the stuff that will grow in phosphate buffer after extended times, but I've not checked into it. I do not use such contaminated buffer in any preps I do.
Has anybody else noticed this, and if so got any idea what it is? Should I add 0.02% sodium azide as a preservative?
Thanks for any insight you can give,
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
We routinely use cacodylate buffer for glutaraldehyde and osmium because it gives a finer grain than phosphate.
Do remember that if you use uranyl acetate en bloc, you need to wash out the cacodyoate or the phosphate before adding the uranium because it is not soluble, and small precipitates will make the image very grainy. We use uranyl acetate in veronal acetate buffer; thus, we wash once in phosphate buffer after osmium, then twice (15-20 min each wash) with veronal buffer before adding the stain. Some folks use aqueus uranyl acetate.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} } } } } } } } accumulates a bit of fluffy stuff at the bottom that is a mid-tone } grey in color, but not a solid black. It kinda looks like fungal stuff, } like the } stuff that will grow in phosphate buffer after extended times, but I've not } checked into it. I do not use such contaminated buffer in any preps I do. } } Has anybody else noticed this, and if so got any idea what it is? Should } I add } 0.02% sodium azide as a preservative?
I have found the material to be fungal (light microscopy) in most cases. Sometimes, however, no hyphae may be encountered but simply amorphous material suggesting some chemical reaction (oxidation/reduction) taking place with the arsenate buffer.
I would hesitate to add azide as it is a potent poison and may affect ultrastructure - especially of mitochondria. Also, therte are the dangers of pouring azide down the drain (formation of explosive lead azide with plumbing solder).
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Renee Recker asks ... } } On 6/8/98 Doug Cromey wrote: } } regarding ... } "Photography in the Age of Falsification" } by: Kenneth Brower, Atlantic Monthly, May 1998 } http://www.theatlantic.com/issues/98may/photo.htm } } 6/9/98 Reply: } } ... I color microscans professionally and can easily } understand the article's application to microscopy. } } 1-when does a digitally manipulated image become a } fake. } 2-how far do you go to satisfy the public's appetite } for exciting images. } 3-artistic expression - adding and subtracting from the } image. } 4-should a picture be labeled as digitally enhanced. } 5-digital capabilities, is it progress or over the top. } } Evidently when ethics are discussed, philosophical gray } areas arise. ...
If I were to condense the article and the points you make (... or maybe its the questions you raise ...), it would come down to the layman's perceptions of "photography" ... and the degree we are all ignorant. I think we all tend to put too much weight on photography being a "window on reality". It is true that many photographer's have given us accurate photos and have truly documented reality. On the other hand, because the digital darkroom has provided more tools, doesn't mean the ethical issues are all of a sudden more important ... after all, the "unsharp mask" was originally created in the darkroom. It has always been possible to remove complexity, if the photographer wanted the viewer to focus on a specific object. And, it has always been possible to remove specific objects and skew a population. Therefore, one point is that it is easier now, but digital tools are not the problem. If I wanted to make any point in particular it is that our faith in photography should be shaken only enough to ask questions. That having been said, it therefore follows the photographer be available for questions. It is interesting in this context how much more easily we can communicate with computer tools, but I don't necessarily mean photographers distribute their e-mail addresses with all their work (... altho that is one aspect of making yourself available ...). A more important aspect of being available for questions is simply accepting other peoples' doubts because photography is NOT reality ... that is, don't take it personally if someone questions your window on reality.
... my $0.02 :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
I am interested in purchasing a cryo-transfer stage for our JEOL 6300F SEM and as far as I know there are just two major manufacturers: Oxford and Emitech. As well as your valued comments any information regarding the interchangability of such systems with other microscopes would be appreciated.
Bugs *will* grow in cacodylate, particularly fungus. You can add a minute pinch of sodium azide to prevent unwanted growth in any buffer. We just throw in a few grains into 1 liter--like about a 1 mm pile on the end of a small spatula. If you need to measure to check your pile the first time, the azide should be about 0.002 M.
Remember this stuff is toxic to people as well as bugs, don't eat it!
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
This question might have been addressed previously, but unfortunately there doesn't seem to be a CGI-based search engine for the MSA archives and there isn't time to download them all and search with a text processor.
I'm doing physiological studies of cultured cells using an inverted microscope. The cells are plated on a coverslip which forms the bottom of a laminar flow chamber. The upper part of the chamber is plastic (polycarbonate, I believe) and the coverslip is sealed to it using a small bead of vacuum grease. At the end of my experiments I need to perform Ab identification of the cells that were measured using ion indicator dyes. I'd like to apply the antibodies (primary and secondary; for intracellular as well as extracellular antigens) in the chamber immediately after the experiment and record the Ab-labelled image for comparison with the ion indicator dye images. I don't want to use aldehydes to fix the tissue because I'm worried about residual aldehyde contamination of the chamber. I'm aware of several options, each with drawbacks:
1. Plate the cells on CELLocate coverslips from Eppendorf (or the equivalent from Bellco), then at the end of the experiment remove the coverslip from the chamber, fix as usual in aldehydes, apply Abs, etc. and relocate the same field of cells using the coverslip coordinates. The problem is that the amount of coverslips sold per minimum order is probably far more than I'd ever need, and they're rather expensive. Does anyone know of another supplier? Alternatively, I heard that Leica once made a small diamond knife mounted to a microscope objective "blank"; this could be moved into place underneath a coverslip by rotating the lens turret and then raised until it just touched the glass. The knife tip was slightly off-center, and it could be rotated by turning the collar on the barrel of the "objective" to inscribe a circle on the coverslip. Does anyone know about this device, or whether something like it is still available?
2. At the end of the experiment fix the cells in the chamber using cold methanol or acetone, apply Abs, etc. One problem I would anticipate is fairly severe shrinkage of the cells with these fixatives, probably to the point where a 1-to-1 image comparison would involve a degree of imagination on the part of a skeptical viewer. I'm not concerned with the ultrastructure of the tissue, which I know can be adversely affected by methanol or acetone. Several questions come to mind, however. I assume that using methanol or acetone would avoid problems of contaminating the plastic chamber for subsequent use with live tissue, but this might be erroneous; any comments? How might I keep the coverslip cold during the period of fixation on the microscope stage? Is it necessary to maintain a low temperature, or can the tissue come to room temperature during fixation (about 15 minutes)? Note: It would be important not to move the stage during the procedure.
Any alternative ideas for solving this problem would be greatly appreciated.
} 2. Micro organisms do not grow in cacodylate buffer as they do in } phosphate buffer.
SNIP!
One would certainly think so. However, I've noticed that 0.2 M caco. buffer, pH=7.0, after long storage times (I'm looking at a bottle of 100 ml mixed up two years ago) accumulates a bit of fluffy stuff at the bottom that is a mid-tone grey in color, but not a solid black. It kinda looks like fungal stuff, like the stuff that will grow in phosphate buffer after extended times, but I've not checked into it. I do not use such contaminated buffer in any preps I do.
Has anybody else noticed this, and if so got any idea what it is? Should I add 0.02% sodium azide as a preservative?
Thanks for any insight you can give,
Gib Ahlstrand
There is a short article published in Stain Technology, vol. 55 #3,1980, pp 191-192, that states that the fungus is Penicillium stoloniferum, a common labatory contaminant. Doug Bray
{P} } 2. Micro organisms do not grow in cacodylate buffer as they do in {BR} } phosphate buffer.
{P} SNIP!
{P} One would certainly think so. However, I've noticed that 0.2 M caco. buffer, {BR} pH=7.0, after long storage times (I'm looking at a bottle of 100 ml mixed up two {BR} years ago) accumulates a bit of fluffy stuff at the bottom that is a mid-tone {BR} grey in color, but not a solid black. It kinda looks like fungal stuff, like the {BR} stuff that will grow in phosphate buffer after extended times, but I've not {BR} checked into it. I do not use such contaminated buffer in any preps I do.
{P} Has anybody else noticed this, and if so got any idea what it is? Should I add {BR} 0.02% sodium azide as a preservative?
{P} Thanks for any insight you can give,
{P} Gib Ahlstrand {BR}
{P} There is a short article published in Stain Technology, vol. 55 #3,1980, pp 191-192, that states that the fungus is {I} Penicillium stoloniferum {/I} , a common labatory contaminant. {BR} Doug Bray {/HTML}
I just wanted to thank all the people who wrote and warned me about ozone generators. The majority recommended better ventilation or use of a HEPA and charcoal activated filter system. I'd like to share with you some of the information that I received: ***** I just read your post. Do not use an ozone generator in a room you are occupying! They generate toxic levels of ozone, which if you are already somewhat sensitive to chemicals can make you worse. They should only be used to kill mold or destroy other alergens in an unoccupied room, then turned off and the room vented well before being occupied. To run while you are in a lab or darkroom, I'd use a charcoal and HEPA filter such as an Envirocaire or Austinaire that I use in my office and bedroom respecively. Cost about $300-400 for a good one, or $3000 for a whole-lab air purification system. ***** Ozone generators are a health risk. Please refer to the following link http://www.epa.gov/iaq/pubs/ozonegen.html"} EPA- "Ozone generators that are sold as air cleaners" ***** Do not use anything that would create OZONE in a darkroom. I worked with photographic based geophysical plotting systems back in the 80's and some people placed OZONE producing equipment in the darkrooms. This OZONE would cause the silver on boards to tarnish and cause circuit falured in equipment placed within the rooms.
Try HEPA filters first and also do not have airducts blowing directly across the top of equipment. The standard difusers did not work properly and some people would place shields at the bottom of the air difusors so the air would blow toward the walls... ***** I'm doubtful about ozone; granted that you will probably not approach the levels once often found in Los Angeles smog, I would prefer to avoid introducing it intentionally. You might have a little sulfur dioxide from the fixer, and the main thing ozone would do would be to convert it to sulfur trioxide, which becomes sulfuric acid.
Effective ventilation would probably be less expensive than good HEPA filtration, and a lot more helpful. ***** Ozone generator? We have a problem with excess ozone on our building (high voltage units, fluorescent ballasts, etc) rotting everything made of rubber (pipette bulbs, tubing, latex gloves, you name it).
Best bet for sensitivity to chemicals is to have an exhaust system (like the hood over a cooking range) to completely remove the chemicals rather than trying to oxidize them. If you are alergic to powdered chemicals, the HEPA might help but fumes will not be removed without using activated charcoal. If you have a unit with both HEPA and activated charcoal, that would do it. ***** Again, thanks! I hope to see many of you at MSA! best regards, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Position available for a 3D Light Microscopist. This individual will oversee the day-to-day operation of a microscopy facility housed in the Laboratory of Receptor Biology and Gene Expression within the National Cancer Institute. The facility will include a confocal microscope, a deconvolution-based microscope and a third instrument yet to be decided, possibly a two-photon microscope or a high-speed low-light level imaging system. The successful candidate will become involved in collaborative research activities with users of this instrumentation, and will play a key role in future decisions to upgrade the facility. Responsibilities of the position will include the assistance and training of users, preventive maintenance of the equipment, and organization of the facility. Candidates must have an ability to work well with biologists and should possess superior organizational skills. Hands-on experience in either confocal or deconvolution microscopy or both is highly desirable. Experience with computers and image processing is also a plus. Candidates with a Ph.D. are preferred. The position is permanent civil service, GS 11/12, $39-61K annual salary. Reply to this message (mcnallyj-at-dce41.nci.nih.gov) with inquiries.
{fontfamily} {param} Times {/param} {bigger} {bigger} {bigger} POSTDOCTORAL POSITIONS IN INTERFACE PHYSICS
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
{/bigger}
Three postdoctoral positions are available in the Interface Physics Group at the University of Illinois at Chicago (UIC). Research in the Interface Physics Group focuses on the use atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale.=20 Current research programs involve ceramics, high-Tc superconductors and optoelectronic/high-power semiconducting materials and devices. The experimental facilities to perform this research are comprehensive: a JEOL 2010 Field-Emission STEM/TEM featuring a 1.4=C5 probe size, "drift free" stage, high-angle annular dark-field detector (Z-contrast), Gatan Imaging Filter, and Noran EDS; a VG HB501A Field-Emission dedicated STEM with EDS, EELS and Auger spectrometers; a JEOL 3010 conventional TEM with digital imaging capabilities and EDS; a JEOL 6320 =46ield-Emission SEM with EDS and Cathodoluminescence; a JEOL JXA733 microprobe; and a Topometrix AFM/STM. In addition to the electron microscopes, specimen preparation facilities include a Gatan Duo-mill, =46ischione precision ion-mill, SouthBay plasma cleaner and Leica Ultramicrotome. The Interface Physics Group has a Silicon Graphics R10000 Power Indigo workstation with a Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The physics department has additional workstations and access to the UIC Convex Exemplar Supercomputer and the National Center for Supercomputing Applications at UIUC. The three research positions are as follows:
(I) Atomic Resolution Analysis of Defects in Ionic Conductors
This position is a joint postdoctoral appointment with Professor Susanne Stemmer in the Department of Physics at UIC. Research performed by the successful candidate for this position will involve the investigation of grain boundaries and defect structures in ionic and mixed ionic/electronic conducting oxide ceramics. The aim of the program is to incorporate experimental results into comprehensive atomic scale models for ionic/electronic transport in these materials.=20 It is anticipated that this position will involve a significant amount of industrial collaboration.
(II) Atomic Resolution Studies of Defects in GaN
This position is a joint postdoctoral appointment with Dr Stephen Pennycook in the Solid State Division at Oak Ridge National Laboratory (ORNL). The successful candidate for this position will be based at UIC to make full use of the extensive MBE and film characterization facilities, but have access to the 300kV VG STEM at ORNL with its 1.26 =C5 probe and soon to be installed PEELS system for imaging and spectroscopy of dislocation core structures. The main aim of this work is to develop an atomic scale understanding of the nucleation mechanisms and electronic properties of dislocations in GaN and related materials. =20
(III) Grain Boundaries in High-Tc Superconductors
This position is a joint postdoctoral appointment with Dr Stephen Pennycook in the Solid State Division at Oak Ridge National Laboratory (ORNL). The successful candidate for this position will be based at ORNL to make full use of the extensive facilities for fabrication and characterization of high-Tc materials. The main aim of this research is to investigate the structure-property relationships at grain boundaries and defects in YBCO, correlating atomic resolution imaging and EELS with macroscopic transport properties. This will involve close collaboration with growth groups in the Solid State Division and focus on two areas: acceptor/donor doped grain boundaries and defects in the RABiTs materials.
Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevent materials issues and an ambition to be part of a developing program pushing at the frontiers of interface physics. Please send a resume and publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. =20 However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Positions are for one year initially, normally renewed for a second year with possibilities existing for further years. Salary is commensurate with experience.=20 UIC is an equal opportunity employer.
{fontfamily} {param} Times {/param} {bigger} {bigger} {bigger} POSTDOCTORAL POSITIONS IN INTERFACE PHYSICS
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
{/bigger}
Three postdoctoral positions are available in the Interface Physics Group at the University of Illinois at Chicago (UIC). Research in the Interface Physics Group focuses on the use atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale.=20 Current research programs involve ceramics, high-Tc superconductors and optoelectronic/high-power semiconducting materials and devices. The experimental facilities to perform this research are comprehensive: a JEOL 2010 Field-Emission STEM/TEM featuring a 1.4=C5 probe size, "drift free" stage, high-angle annular dark-field detector (Z-contrast), Gatan Imaging Filter, and Noran EDS; a VG HB501A Field-Emission dedicated STEM with EDS, EELS and Auger spectrometers; a JEOL 3010 conventional TEM with digital imaging capabilities and EDS; a JEOL 6320 =46ield-Emission SEM with EDS and Cathodoluminescence; a JEOL JXA733 microprobe; and a Topometrix AFM/STM. In addition to the electron microscopes, specimen preparation facilities include a Gatan Duo-mill, =46ischione precision ion-mill, SouthBay plasma cleaner and Leica Ultramicrotome. The Interface Physics Group has a Silicon Graphics R10000 Power Indigo workstation with a Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The physics department has additional workstations and access to the UIC Convex Exemplar Supercomputer and the National Center for Supercomputing Applications at UIUC. The three research positions are as follows:
(I) Atomic Resolution Analysis of Defects in Ionic Conductors
This position is a joint postdoctoral appointment with Professor Susanne Stemmer in the Department of Physics at UIC. Research performed by the successful candidate for this position will involve the investigation of grain boundaries and defect structures in ionic and mixed ionic/electronic conducting oxide ceramics. The aim of the program is to incorporate experimental results into comprehensive atomic scale models for ionic/electronic transport in these materials.=20 It is anticipated that this position will involve a significant amount of industrial collaboration.
(II) Atomic Resolution Studies of Defects in GaN
This position is a joint postdoctoral appointment with Dr Stephen Pennycook in the Solid State Division at Oak Ridge National Laboratory (ORNL). The successful candidate for this position will be based at UIC to make full use of the extensive MBE and film characterization facilities, but have access to the 300kV VG STEM at ORNL with its 1.26 =C5 probe and soon to be installed PEELS system for imaging and spectroscopy of dislocation core structures. The main aim of this work is to develop an atomic scale understanding of the nucleation mechanisms and electronic properties of dislocations in GaN and related materials. =20
(III) Grain Boundaries in High-Tc Superconductors
This position is a joint postdoctoral appointment with Dr Stephen Pennycook in the Solid State Division at Oak Ridge National Laboratory (ORNL). The successful candidate for this position will be based at ORNL to make full use of the extensive facilities for fabrication and characterization of high-Tc materials. The main aim of this research is to investigate the structure-property relationships at grain boundaries and defects in YBCO, correlating atomic resolution imaging and EELS with macroscopic transport properties. This will involve close collaboration with growth groups in the Solid State Division and focus on two areas: acceptor/donor doped grain boundaries and defects in the RABiTs materials.
Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevent materials issues and an ambition to be part of a developing program pushing at the frontiers of interface physics. Please send a resume and publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. =20 However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Positions are for one year initially, normally renewed for a second year with possibilities existing for further years. Salary is commensurate with experience.=20 UIC is an equal opportunity employer.
We have need for a double tilt - low background - cold stage for a Philips EM430 TEM. If anyone knows of one that someone is willing to part with, for free or otherwise, please contact me.
Thanks.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor Mechanical, Materials & Aerospace Engineering University of Central Florida 4000 Central Florida Blvd., PO Box 162450 Orlando, FL 32816-2450 phone: 407 823-5770,fax: 407 823-0208 lag-at-ucf.edu
Can you help us? A student is growing very small colonies of cells and she wishes to view their ultrastructure. There are about 50 cells per colony and about 4 colonies per 3cm polystyrene petri dish. Our problem is that the colonies are growing between two layers of agar. The bottom layer is 0.5% agar in PBS and the top layer is 0.33% agar in PBS. The colonies break up and float away during processing and the agar just moves around. The colonies are not attached/embedded in the agar. We have tried (1)cutting around the colonies and sucking the whole lot up (agar + colony) and treating it as a pellet but the cells are impossible to find in the resin, and (2)we have tried to just gently fix the mass and process it as a whole but we end up with no cells as the colony breaks up and the cells disperse. Is there anyone out there who could offer a suggestion. Thanks
Sarah Ellis
Research Division Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne, Victoria 3000 Australia
Dear All, One of our users intends to do image analysis on color images, taken from histological sections, immunostained with DAB together with an other chromogen like for instance NBT or Fast Red. Since we are aware of the fact that we may encounter problems when comparing images taken at different days or even at different time points during a session (lamp-intensity and -color fluctuations, exposure time etc., etc.) we would welcom ANY comments and suggestions from people already doing this kind of analysis. ALL information on: * the camera system used * (automatic) correction for possible differences in imaging conditions * the image analysis software used * and other points which may be important is greatly appreciated. Many thanks in advance. Lauran Oomen *****------------------------**********----------------------------***** Lauran Oomen, The Netherlands Cancer Institute, Dept. Biophysics (H-0) Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands oomen-at-nki.nl, TEL +31-20-5121898, FAX +31-20-5121893 WWW page: http://www.nki.nl/nkidep/biofys/microscopy/digmiclo.htm
Paul Geroir asked: } I am interested in purchasing a cryo-transfer stage for our JEOL 6300F } SEM and as far as I know there are just two major manufacturers: Oxford } and Emitech. As well as your valued comments any information regarding } the interchangability of such systems with other microscopes would be } appreciated.
In addition to Oxford and Emitech, VG Microtech manufactures an SEM cryopreparation system as part of its "Polaron Range" of EM specimen preparation instrumentation. If any of you are interested in additional information, please e-mail me your postal address, and I will be pleased to send you current literature on this product.
One of these systems is currently resident in the applications lab at JEOL UK. I will be organizing a cryo-preparation seminar at JEOL USA later this year, and will let the group know the details once they have been firmed up.
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
POSTDOCTORAL POSITION - RENAL CELL PHYSIOLOGY I have an opening for a Postdoctoral Fellow in my laboratory available after October 1, 1998. The research is concerned with the cellular and molecular biology of renal xenobiotic excretion. We use a variety of techniques including, isolated renal proximal tubules, renal cells in culture, fluorescent substrates and confocal microscopy to follow substrates across epithelial cells and to characterize the processes involved and the signal transduction pathways regulating transport. Candidates should have a Ph.D. or M.D. degree, less than 5 years postdoctoral experience and a background in cellular physiology, membrane transport or renal physiology. This is a non-tenure track position (NIH IRTA Fellow).
_____________________________ Dr. David S. Miller Laboratory of Pharmacology & Chemistry NIH/NIEHS Research Triangle Park, NC 27709
I have an old brass microscope, with two eyepieces and three objectives. I have had it since my childhood and my memory is that my father had used it in college/high school, in Seattle c. 1916-1924, and that it was second hand at that time. Since he was an economist, he did not use it professionally (no, he was not a microeconomist). The microscope has engraved on it's base, in large letters, "A. S. Aloe & Co, St. Louis, Mo." It also has, in a different font, "Diagnostician," which I take to be the model name of the instrument. There are no other identifying marks that I can discern on the body/base. The eyepieces and two of the objectives are stamped "A.S. Aloe," the third objective is inscribed "Gundlach Optical Co, Rochester N.Y." Because this third objective appears to be of the same form/fit/function/material/finish as the other two, my suspicion is that Aloe was a distributor and Gundlach was the manufacturer, but the evidence is weak. I would greatly appreciate any suggestions for tracing the history of this instrument, and particularly if there is any record of an A.S. Aloe & Co. in St. Louis, and a Gundlach Optical Co. in Rochester, both in the late 1800's or early in this century. Thx for your attention, and help.
Robert. F. Rowntree
Bob & Esther Rowntree 808 Murray Ave San Luis Obispo, CA 93405 (805)549-9107
[skip] } It has always been } possible to remove complexity, if the photographer wanted the viewer to } focus on a specific object. And, it has always been possible to remove } specific objects and skew a population.
Not to mention that by varying the f-stop, speed, or lighting conditions the original photograph can be chosen to highlight what the photographer wants. Yours, Bill Tivol
I routinely use an image analysis system to do exactly what you describe below. So here are the specifics:
* the camera system used For the last 6 years I've used the Sony DXC-151 CCD/RGB camera which exports the image directly into the frame grabber board in my computer. It should be noted that the system I use is compatible with just about any kind of camera system and I often work with images that were made with other camera systems (both digital and video) which are on diskettes in tiff or jpeg format.
* (automatic) correction for possible differences in imaging conditions Corrections for different conditions can be made either directly after the image is 'grabbed' or while setting the color thresholds, most of the time I've found that it's really not necessary.
* the image analysis software used BioQuant TCW by: R&M Biometrics 5611 Ohio Ave. Nashville, TN 37209 615-350-7866
They have a web site, search for bioquant.com
Cost (including computer) is under US$30,000
* and other points which may be important Make sure that the system you get is open ended enough so that other applications such as densitometry, stenography, area and line measurements etc. can also be performed. You might be surprised at how much you will want to use these additional applications. Also, it should be relatively simple to transfer the collected data to a spreadsheet. BQ does all of the preceding.
No Suggestions I am simply a very satisfied customer and have no financial or business interest in R&M Biometrics
-- Begin original message --
Dear All, One of our users intends to do image analysis on color images, taken from histological sections, immunostained with DAB together with an other chromogen like for instance NBT or Fast Red. Since we are aware of the fact that we may encounter problems when comparing images taken at different days or even at different time points during a session (lamp-intensity and -color fluctuations, exposure time etc., etc.) we would welcom ANY comments and suggestions from people already doing this kind of analysis. ALL information on: * the camera system used * (automatic) correction for possible differences in imaging conditions * the image analysis software used * and other points which may be important is greatly appreciated. Many thanks in advance. Lauran Oomen *****------------------------**********----------------------------***** -- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
Dear Sarah, } } Can you help us? A student is growing very small colonies of cells and } she wishes to view their ultrastructure. There are about 50 cells per } colony and about 4 colonies per 3cm polystyrene petri dish. Our problem } is that the colonies are growing between two layers of agar. The bottom } layer is 0.5% agar in PBS and the top layer is 0.33% agar in PBS. The } colonies break up and float away during processing and the agar just } moves around. The colonies are not attached/embedded in the agar. } We have tried (1)cutting around the colonies and sucking the whole lot } up (agar + colony) and treating it as a pellet but the cells are } impossible to find in the resin, and (2)we have tried to just gently } fix the mass and process it as a whole but we end up with no cells as } the colony breaks up and the cells disperse. } Is there anyone out there who could offer a suggestion.
I am most certainly not an expert on this, but could you try cut- ting around the colonies, cooling the dish to 0 C (to harden the agar, but not freeze the cells or medium), then extract the agar + cells? If this gives you the intact colony between the agar layers, you could then trim away some of the agar. Good luck. Yours, Bill Tivol
Gernot, Cacodylate is probably one of the most cytoplasmic extracting buffers where as pbs or phosphate is better but must be replaced before osmication. My rule is PBS or phosphate buffers for immuno-EM and cacodylate for standard fixations. Check out Hyat or Glauret's text series on fixation/buffers.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Dear Sarah, } } } } Can you help us? A student is growing very small colonies of cells and } } she wishes to view their ultrastructure. There are about 50 cells per } } colony and about 4 colonies per 3cm polystyrene petri dish. Our problem } } is that the colonies are growing between two layers of agar. The bottom } } layer is 0.5% agar in PBS and the top layer is 0.33% agar in PBS. The } } colonies break up and float away during processing and the agar just } } moves around. The colonies are not attached/embedded in the agar. } } We have tried (1)cutting around the colonies and sucking the whole lot } } up (agar + colony) and treating it as a pellet but the cells are } } impossible to find in the resin, and (2)we have tried to just gently } } fix the mass and process it as a whole but we end up with no cells as } } the colony breaks up and the cells disperse. } } Is there anyone out there who could offer a suggestion. } } I am most certainly not an expert on this, but could you try cut- } ting around the colonies, cooling the dish to 0 C (to harden the agar, but } not freeze the cells or medium), then extract the agar + cells? If this } gives you the intact colony between the agar layers, you could then trim } away some of the agar. Good luck. } Yours, } Bill Tivol
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
We are looking to upgrade our Cambridge S250 SEM with a slow scan image digitiser. Specifically we are looking for a system to grab the image from the photo tube at fairly high resolution (at present we can only grab at video res from the internal digital image store which does not work for BSE images). I do not think we can afford a system which controls the beam scan itself - besides this is probably beyond our requirements.
If anybody has recently acquired such a system and has an opinion on what is best or how to go about it I would be grateful.
Please reply to me direct and I will provide the list with a summary.
Many thanks,
Stu
P.S. This morning's headline in the Independent newspaper in the UK:
"...at 4:30pm today a man will kick a ball" ---------------------- Stuart Kearns stuart.kearns-at-bris.ac.uk
Dear Friends, I was disappointed last year when Victawet, the release agent I used on my carbon evaporator, was taken off the market because the manufacturers didn't want to deal with the safety labeling issues, so I was told. I had found Victawet a very effective release agent, and I have to clean my bell jar quite frequently in our busy SEM/EDX lab. I have been using the spray-on release agents since then and I find them expensive and very ineffectual, making the bell-jar cleaning process a long, hard scrub. Last month, in frustration, I lathered up my hands with the bar soap I keep at my sink and smeared it inside my bell-jar. I figured it couldn't be worse than the spray-on. It worked beautifully! Yesterday I wiped it with a wet sponge and everything lifted off with no effort. It does not seem to affect the high vaccuum performance and you can't beat the price. Makes me wonder what I have been spending all my scarce money on all this time.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
To all Midwest microscopists in Central States or MIKMAS societies.
The Ballot sent out this week had an incorrect fax #.
The correct Fax # for Lou Ann is: 217-244-1652
Thanks,
Lou Ann
***************************************-at-redfoot Lou Ann Miller Center for Microscopy & Imaging College of Veterinary Medicine Dept. of Veterinary Biosciences University of Illinois Rm 1108 Basic Sciences Bld 2001 S Lincoln Ave. Urbana, Illinois 61802
We are looking to upgrade our Cambridge S250 SEM with a slow scan image digitiser. Specifically we are looking for a system to grab the image from the photo tube at fairly high resolution (at present we can only grab at video res from the internal digital image store which does not work for BSE images). I do not think we can afford a system which controls the beam scan itself - besides this is probably beyond our requirements.
If anybody has recently acquired such a system and has an opinion on what is best or how to go about it I would be grateful.
Please reply to me direct and I will provide the list with a summary.
Many thanks,
Stu
P.S. This morning's headline in the Independent newspaper in the UK:
"...at 4:30pm today a man will kick a ball"
---------------------- Stuart Kearns stuart.kearns-at-bris.ac.uk
Data Translation (http://www.datx.com) sells a slow scan board, the DT3152. It accepts just about any video signal. Setting it up for analog X-Y rasters can be a chore but the images (up to 4Kx4K I believe) are very good.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
You might try first infiltrating the cells and agar in situ with a protein like bovine serum albumin (or possibly gelatin) and then fixing the entire petri dish contents by gently overlaying some 4% glutaraldehyde. The glut should not mix with the albumin but should layer on top of it. Allow this to stand for several hours. The glut will diffuse into the albumin, cross link it to a firm configuration that you will be able to cut with a razor blade into cubes. You will probably have to re-fix the blocks that you initially trim out to firm them up even more. An additional hour in glut should do this.
You should experiment with this first to get the times and proper concentrations of protein, but we used it successfuly a number of years ago. You might even use microwaving to accelerate the fixation.
A second possibility would be to do a freeze substitution fixation. But this will be more complicated. Try the easier one first.
Keep us all informed as to your results.
} Can you help us? A student is growing very small colonies of cells and } she wishes to view their ultrastructure. There are about 50 cells per } colony and about 4 colonies per 3cm polystyrene petri dish. Our problem } is that the colonies are growing between two layers of agar. The bottom } layer is 0.5% agar in PBS and the top layer is 0.33% agar in PBS. The } colonies break up and float away during processing and the agar just } moves around. The colonies are not attached/embedded in the agar. } We have tried (1)cutting around the colonies and sucking the whole lot } up (agar + colony) and treating it as a pellet but the cells are } impossible to find in the resin, and (2)we have tried to just gently } fix the mass and process it as a whole but we end up with no cells as } the colony breaks up and the cells disperse.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We're considering the feasibility of purchasing a new transmission scope and are gathering info to decide initially which styles and makes of scopes would be most appropriate for our needs. We currently have a Phillips 300 which works beautifully (thanks to Mark Huller, our excellent local Phillips service tech), however, we've become aware that replacement parts are quickly becoming unavailable in the event of a major repair. Since the institute is planning construction of a new research building, it only makes sense to plan the purchase of a new scope as well.
To date, all our specimens have been biological in nature and analyses involving EELS, X-ray, diffraction, etc are not likely (but not outside the realm of possibility). Usage isn't expected to be particularly heavy (although that is also certainly subject to change). We've made the usual paper prints from our negatives but more recently have simply scanned the negs and "processed" them using Adobe Photoshop. Ideally, should the technology be adequate, we would like the capability to electronically capture the images directly from the scope as well as continue with conventional darkroom techniques.
Should anyone have any suggestions or recommendations with regards to manufacturers, models, and imaging capability and quality, I'd be grateful. Communications may be sent directly to me.
Thanks,
Jaclynn M. Lett, Research Assistant
jmlett-at-cid.wustl.edu
Central Institute for the Deaf Center for the Study of the Biology of Hearing and Deafness 818 S. Euclid Ave. St. Louis, MO 63110
Stuart Kearns wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Folks, } } We are looking to upgrade our Cambridge S250 SEM with a } slow scan image digitiser. Specifically we are looking for } a system to grab the image from the photo tube at fairly } high resolution (at present we can only grab at video res } from the internal digital image store which does not work } for BSE images). I do not think we can afford a system } which controls the beam scan itself - besides this is } probably beyond our requirements. } } If anybody has recently acquired such a system and has an } opinion on what is best or how to go about it I would be } grateful. } } Please reply to me direct and I will provide the list with } a summary. } } Many thanks, } } Stu } } P.S. This morning's headline in the Independent newspaper } in the UK: } } "...at 4:30pm today a man will kick a ball" } ---------------------- } Stuart Kearns } stuart.kearns-at-bris.ac.uk I've been evaluting GW's "printerface". Works great for the money.
I think you have wasted your money by buying in the past special cleaning agents. I have been in the EM field for more years than I like to remember (} 35 years). I have never used any special cleaning agents and clean the bell yar of the evaporator after usage with alcohol and paper towels. I experienced no problems and the bell yar is as clean as it was when we bought the unit.
Hans Brinkies Swinburne, University of Technology School of Engineering and Science P.O.Box 218 HAWTHORN. Vic. - 3174 - Melbourne, Australia
they have a very good section on historical microscopes.
Hans Brinkies Swinburne, University of Technology School of Engineering and Science P.O.Box 218 HAWTHORN. Vic. - 3174 - Melbourne, Australia Hans G Brinkies Senior Lecturer Swinburne, University of Technology School of Engineering and Science Electron Microscopy Hawthorn, 3122, Melbourne - Australia
Funny that Mary. I've been applying a thin coating of detergent, well dried before use, on vacuum bell jars for about 30 years. Neither did I invent that particular wheel: The Girl Guides have used soap/detergent on the outside of saucepans to help with the clean-up for at least 60 years. Which Journal would publish such a simple gem? Thanks to the microscopy server. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} Dear Friends, } I was disappointed last year when Victawet, the release agent I used on my } carbon evaporator, was taken off the market because the manufacturers didn't } want to deal with the safety labeling issues, so I was told. I had found } Victawet a very effective release agent, and I have to clean my bell jar } quite frequently in our busy SEM/EDX lab. I have been using the spray-on } release agents since then and I find them expensive and very ineffectual, } making the bell-jar cleaning process a long, hard scrub. Last month, in } frustration, I lathered up my hands with the bar soap I keep at my sink and } smeared it inside my bell-jar. I figured it couldn't be worse than the } spray-on. It worked beautifully! Yesterday I wiped it with a wet sponge and } everything lifted off with no effort. It does not seem to affect the high } vaccuum performance and you can't beat the price. Makes me wonder what I } have been spending all my scarce money on all this time. } } Regards, } Mary } } Mary Mager } Electron Microscopist } Metals and Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } fax: 604-822-3619 } e-mail: mager-at-interchange.ubc.ca } }
Mary Mager wrote: =============================================== I was disappointed last year when Victawet, the release agent I used on my carbon evaporator, was taken off the market because the manufacturers didn't want to deal with the safety labeling issues, so I was told. I had found Victawet a very effective release agent, and I have to clean my bell jar quite frequently in our busy SEM/EDX lab. I have been using the spray-on release agents since then and I find them expensive and very ineffectual, making the bell-jar cleaning process a long, hard scrub. Last month, in frustration, I lathered up my hands with the bar soap I keep at my sink and smeared it inside my bell-jar. I figured it couldn't be worse than the spray -on. It worked beautifully! Yesterday I wiped it with a wet sponge and everything lifted off with no effort. It does not seem to affect the high vacuum performance and you can't beat the price. Makes me wonder what I have been spending all my scarce money on all this time. ================================================== I can assure you that Victawet® is very much alive and well, or at least it is here at SPI where we have what we project to be a lifetime supply. And so far as I know, it is also available from our competitors, such as Ladd Research and perhaps from others. There are different "grades" and there are different ways of preparing it, so the end product itself might not be the same in appearance.
You can find Viactawet listed on our website.
This product is indeed one of those items that once you use it, it is almost impossible to imagine life with out it. And for the price of a 5 gm vial, which is not that much more in cost than that of several bars of soap, you get what would be a near life time supply for many laboratories.
Your point that there are alternatives to Victawet is well taken. But there are other applications, such as serving as a release agent for carbon films and Pt/C replicas for which so far as I know there are no alternatives.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I need the instruction manual for the Cryo-ultra mocrotomes - LKB I and - LKB II Does anybody know where I can get them? Unfortunately, the company LKB does not exist any more, so I do not know who I can contact. Any help will be highly appriciated. Thank you very much in advance.
Manuela
---------------------- Manuela Finke University of Bristol Department of Oral and Dental Science Dental Materials and Biomaterials Group Lower Maudlin Street Bristol BS1 2LY tel:0117/9284537 fax:0117/9284780 e-mail:M.Finke-at-bris.ac.uk
We have need for a double tilt - low background - cold stage for a Philips EM430 TEM. If anyone knows of one that someone is willing to part with, for free or otherwise, please contact me.
Thanks.
************************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor Dept. of Mechanical, Materials, and Aerospace Eng.
Director, Cirent/UCF Materials Characterization Facility President, Florida Society for Microscopy
University of Central Florida phone (407) 823-5770 PO Box 162450 fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA MCF (407) 275-4354 ----------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain *************************************************************************
I need the instruction manual for the Cryo-ultra mocrotomes - LKB I and - LKB II Does anybody know where I can get them? Unfortunately, the company LKB does not exist any more, so I do not know who I can contact. Any help will be highly appriciated. Thank you very much in advance.
Manuela
---------------------- Manuela Finke University of Bristol Department of Oral and Dental Science Dental Materials and Biomaterials Group Lower Maudlin Street Bristol BS1 2LY tel:0117/9284537 fax:0117/9284780 e-mail:M.Finke-at-bris.ac.uk
I assume that you mean the LKB 14800 cryokit (when you say LKB I). This was the basic grey plastic box without foam insulation and some of the cryo-tools. I have the instructions which I could photocopy and mail to you in a few days time - please let me know.
However I don't know what you mean by cryo-ultramicrotome LKB II - is this just the updated grey box or do you mean the Cryo-Nova? In any event I can't help with that.
Have you tried contacting Leica UK (tel. 01908 666 663) who took over from Cambridge Instruments who took over from Reichert.
Malcolm Haswell Electron Microscopy School of Health Sciences University of Sunderland Sunderland SR1 3SD UK tel 0191 515 2872 ----------
Dear all,
I need the instruction manual for the Cryo-ultra mocrotomes - LKB I and - LKB II Does anybody know where I can get them? Unfortunately, the company LKB does not exist any more, so I do not know who I can contact. Any help will be highly appriciated. Thank you very much in advance.
Manuela
---------------------- Manuela Finke University of Bristol Department of Oral and Dental Science Dental Materials and Biomaterials Group Lower Maudlin Street Bristol BS1 2LY tel:0117/9284537 fax:0117/9284780 e-mail:M.Finke-at-bris.ac.uk
by cap2.capitalnet.com (8.8.5/8.8.5) with SMTP id IAA07709 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 11 Jun 1998 08:09:40 -0400 (EDT) Message-Id: {3.0.2.32.19980611081411.00a39584-at-capitalnet.com} X-Sender: dpurdy-at-capitalnet.com X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.2 (32)
About six months ago, I posted a message on this group indicating I was searching for a used phototube for a Wild M7A or M3Z stereomicroscope. I was also searching for an inclined binocular tube for either the M7A or M3Z.
I am still looking for these two parts. If anyone has a spare unit in good condition they would like to sell please reply to this message with a description, price and your location.
As usual in all scientific work, the integrity, education, background and reputation of the individual generating the images is the bottom line when it comes to "truth" contained within the images, or any other work one does. The characteristics aren't exclusive, a "good" scientist can still make judgmental and emotional errors, but one has to start somewhere. If the images contain information that is clear and well documented I don't see that it is terribly important to know exactly how they were generated ( except possibly from the point of view of replicating the work).
Manuela, LKB was bought out by Leitz (as was Reichert) and they renamed themselves "Leica" - not after the Russian space-dog, but the best early 35mm camera. I believe that Leica, if you find the right person, can and will help. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} } Dear all, } } I need the instruction manual for the Cryo-ultra mocrotomes } - LKB I and } - LKB II } Does anybody know where I can get them? } Unfortunately, the company LKB does not exist any more, so } I do not know who I can contact. } Any help will be highly appriciated. Thank you very much in } advance. } } Manuela } } ---------------------- } Manuela Finke } University of Bristol } Department of Oral and Dental Science } Dental Materials and Biomaterials Group } Lower Maudlin Street } Bristol BS1 2LY } tel:0117/9284537 } fax:0117/9284780 } e-mail:M.Finke-at-bris.ac.uk } } }
Forgive my ignorance, but if the freezing point of isopentane is 113 K and that of propane is 84 K, how come a mixture of these freezes at temperatures below 77 K (boiling LN2)?
Thanks you.
James Wesley-Smith EM Unit University of Natal Durban South Africa
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On June 11, 1998 James Wesley-Smith wrote:
"Forgive my ignorance, but if the freezing point of isopentane is 113 K a= nd
that of propane is 84 K, how come a mixture of these freezes at =
temperatures below 77 K (boiling LN2)?"
Reply:
Mixtures have lower freezing points due to "freezing point depression." =
The familiar example is mixing salt and water to give a brine solution at=
-15 deg C, used for making ice cream.
In the Handbook of Chemistry and Physics, a table shows that depressions of 50 to 100 degrees C are common in mixtures of organic molecules. So, this is likely why isopentane and propane together freeze below 77K, just= 7 degrees below propane.
[If you want to get quantitative, arm yourself with the freezing point depression constant for either of these compounds. You can then calculat= e the freezing point of the mixture. Ref. "Physical Chemistry" by WJ Moore= ]
Cheers,
Nathan Haese Walnut Creek, California Nathan_Haese-at-compuserve.com
The MSA Golf Tourney will be held on July 12 at 9:00 am at the Atlanta International Golf course. Attached is the registration form in Word 97 .doc format. Call Mark Rigler at 800-421-8451 if you need additional info.
Does anybody know of a manufacturer or supplier of magnetic field containment loops. We are having problems with our Hitachi S-800FE SEM - getting possible structurally transmitted magnetic interference from lab two floors up. Our SEM is located in a basement with the structural columns (steel reinforced concrete) directly above and to the side of the SEM. Interference manifests itself in the form of CRT screen "wobble" of set frequency especially noticable at high mag (} x3K). Equipment in other lab determined to be the source of the problem is a Microwave plasma reactor (interference occurs when heater is on) and an Electro-cyclotron resonance (ECR) instrument. Need urgent help.Cheers Dr Alan Templeton Centre for Physical Electronics and Materials SEEIE South Bank University 103 Borough Road, London SE1 0AA TEL 44 171 815 7521 /7571 FAX 44 171 815 7599 email templea-at-sbu.ac.uk
Our laboratory has recently started receiving a few skin biopsies to screen by EM for evidence of Ehlers Danlos disease. This is a new entity for us therefore, we have no guidelines in place for proper evaluation. What are the observations by LM? I have seen examples of the ultrastructural alterations of the collagen fibrils but am wondering how much of the collagen is affected, overall? Any feedback would be greatly appreciated!
Pat
Patrice Abell-Aleff Electron Microscopy Core Facility Mayo Clinic 200 1st Street SW Rochester, Minnesota 55905 e-mail: abellaleff.patrice-at-mayo.edu phone: 507-284-3148 fax: 507-284-9349
For those of you who cannot pull up the MSA registration form in the Word format, I have attached a tif file for your download. Call me if you have a problem at 800-204-6402.
} Forgive my ignorance, but if the freezing point of isopentane is 113 K and } that of propane is 84 K, how come a mixture of these freezes at } temperatures below 77 K (boiling LN2)? } Granted that dissolving a solid like salt in a solution leads to freezing point depression. However, I think the freezing point depression in this case might be better compared to a solution of two miscible liquids. I am used to the phenomenon being discussed with application to metals. A solution of tin and lead is liquid below the freezing points of the individual metals. With a "eutectic" mixture you encounter the maximum freezing point depression. I suppose the same principle applies here, even though the compounds are normally liquids (or even gases) at 'normal' conditions.
Spicer Consulting 10 Wentworth Drive Bedford MK41 8BB UK
Phone (+44)/(0) 234-346419
It was actually sold at the time by Agar in the UK and by Ted Pella in the US.
This was something like 6 years ago, and I cannot vouch for how up-to-date this information is. I can tell you though, that it does work!
Tony Garratt-Reed
At 04:24 PM 6/11/98 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
Hi there, Just a quick note to let everyone know that they should send in their registrations as soon as possible for hotel rooms in Atlanta. For those of you who practice "brinkmanship" and leave everything to the last minute, I would like to warn you that there is an NAACP conference in the city from the 7th to the 19th of July and so the hotel rooms downtown are hard to come by and also there is a fairly large convention at the Merchandize Mart too. I didnt get my hotel resrvations in to the convention bureau and so had to make my own reservations and some of the hotels are saying they are fully booked. This of course doesnt mean that the convention center doesnt have a large block of rooms held for us, but it does mean that you should make reservations asap. If you have troubles the following hotels are near the Georgia World Congress Center:
Atlanta Hilton and Towers The Ritz Carlton Atlanta The Howard Johson Suites The Confort Inn Downtown The Best Western American Hotel
or you can check:
http://www.acvb.com/ac_stay.asp
Just a reminder.....
John F. Mansfield
Note new Area Code (734)
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Bob Lawrence wrote: } I don't see that it is terribly important to know } exactly how they (images) were generated
This may be true in general. However, there is one type of image that should only be published with full information about exactly how it was generated, and that is a simulated HRTEM image. Each such image should have associated with it all the parameter values used to generate it as well as the name and version number of the software used. I am surprised at the number of papers I am called on to referee that do not contain such information. The parameter values should be given to inform the reader of what conditions would be necessary to obtain such an image. The software should be identified in order to allow for any peculiarities of particular software, or even errors that may be revealed later. An early version of SHRLI (Simulated High-Resolution Lattice Images) inverted beam indices in all of its diffraction patterns (SHRLI80), and a version of EMS imposed incident beam convergence at twice the input value (see 51st Proc MSA (1993) 974-975).
-Mike O'Keefe
-------------------------------------------------------------- Bob Lawrence wrote:
} Gentlefolk, } } As usual in all scientific work, the integrity, education, } background and reputation of the individual generating the images is the } bottom line when it comes to "truth" contained within the images, or any } other work one does. The characteristics aren't exclusive, a "good" } scientist can still make judgmental and emotional errors, but one has to } start somewhere. If the images contain information that is clear and } well documented I don't see that it is terribly important to know } exactly how they were generated ( except possibly from the point of view } of replicating the work).
TO: Nancy Zjaba National Semiconductor South Portland, ME
Soft Imaging System GmbH has image analysis software called analySIS. We also have specific modules for grain analysis. If you like more information, please visit our web site http://www.soft-imaging.de or contact us at sales-at-soft-imaging.com or myself and we will be glad to talk to you and send you brochures.
Kuni Tatsumoto kt-at-soft-imaging.com (888) FIND-SIS
Dr. Templeton: Below is the Web site for Spicer Consulting. They make an excellent mag field canceling system. I've used 3 of their units, the most recent purchased just 3 weeks ago.
http://www.spiceruk.co.uk/
Good luck! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 KFAB PFA Lab Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Alan Templeton wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anybody know of a manufacturer or supplier of magnetic field } containment loops. We are having problems with our Hitachi S-800FE } SEM - getting possible structurally transmitted magnetic interference } from lab two floors up. Our SEM is located in a basement with the } structural columns (steel reinforced concrete) directly above and to the } side of the SEM. Interference manifests itself in the form of CRT } screen "wobble" of set frequency especially noticable at high mag } (} x3K). Equipment in other lab determined to be the source of the } problem is a Microwave plasma reactor (interference occurs when } heater is on) and an Electro-cyclotron resonance (ECR) instrument. } Need urgent help.Cheers } Dr Alan Templeton } Centre for Physical Electronics and Materials } SEEIE } South Bank University } 103 Borough Road, London } SE1 0AA } TEL 44 171 815 7521 /7571 } FAX 44 171 815 7599 } email templea-at-sbu.ac.uk
On Thu, 11 Jun 1998, Alan Templeton wrote: Hi Alan,
We have have been very interested in this problem as well. We have had trials of, and heard other good reports of, the field cancelling system sold by Oxford Instruments. Contact them at: Oxford Instruments, Research Instruments, Tubney Wood, Abingdon, Oxon. OX13 5QX. 01865-393200. Ask for Judith Brock. e-mail:- judith.brock-at-oxinst.co.uk
Unfortunately I have no connections or financial interests.
Good luck, Ron } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anybody know of a manufacturer or supplier of magnetic field } containment loops. We are having problems with our Hitachi S-800FE } SEM - getting possible structurally transmitted magnetic interference } from lab two floors up. Our SEM is located in a basement with the } structural columns (steel reinforced concrete) directly above and to the } side of the SEM. Interference manifests itself in the form of CRT } screen "wobble" of set frequency especially noticable at high mag } (} x3K). Equipment in other lab determined to be the source of the } problem is a Microwave plasma reactor (interference occurs when } heater is on) and an Electro-cyclotron resonance (ECR) instrument. } Need urgent help.Cheers } Dr Alan Templeton } Centre for Physical Electronics and Materials } SEEIE } South Bank University } 103 Borough Road, London } SE1 0AA } TEL 44 171 815 7521 /7571 } FAX 44 171 815 7599 } email templea-at-sbu.ac.uk }
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Dear Microscopists, Can anybody tell me where to get urease antibodies? I don't know the origin of the urease (bacterial or fungal), is there an antibody recognizing both? It is for fluorescence immunolokalisation, many thanks, Arthur
PS: anybody knows interesting papers about fungal urease, role of urea etc? I didn't find very much.
Dr. Arthur Schuessler TU Darmstadt FB10 Botanik, AG Kluge Schnittspahnstr. 10 64287 Darmstadt
Integrated Dynamics Engineering 84 Cummings park Woburn, MA 01801 tel: 617-938-5120 fax: 617 938 5122 100125.1130-at-compuserve.com
} Steve Rozeveld } } Dow Chemical Co. } 1897 Bld. Door E-43 } Midland, MI 48667 } sjrozeveld-at-dow.com } % (517) 636-5167 } Fax: (517) 638-6443
} ---------- } From: Ron Doole[SMTP:ron.doole-at-materials.oxford.ac.uk] } Sent: Friday, June 12, 1998 2:08 AM } To: Microscopy Community } Subject: Re: SEM - problems with magnetic interference } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-Just my guess as to what happens. We constanly have problems trying to adjust different Monitors, software, and operating systems to match the same colors when printing out color images. We now do this subjectively, because the color balance seems to be set differently for each system. A color Standard "slide" would be a terrific start, and would at least allow the same starting point and settings to be made on hardware. Although I would expect CRT color-monitor variations over time, I expect the software will interpret color balance with precision each time, until a new version or a different process algorythm is employed. I would be greatful to try this slide. We actually have used the Television production color screen to pre-set our color monitors and synchronize them with a video camera. I am not sure how much the video lens alters the colors. You can see such a color screen on the television networks in early AM. However, even those colors screens fade due to the ambient lighting causing more need for color correction over time. So many factors come into play, resolution, screen display, printer interpretation, digital acquisition, balance, intensity, reflectivity, absorbance, transmittance, black level, that it will be almost impossible to get precise color balance, but it will be much closer than not having one. Tom Baginski Technical Coordinator for Microscopy
In response to: -a good standard available, mounted on a traditional 1"x3" slide, at reasonable price ($35-50), would a "color test plate"
Thanks for the names and addresses of companies selling field containment systems- have got a few now to be getting on with but if you know of any others please let me know - should prove very useful in solving this problem.
Cheers alan templeton. ------------------ Dr Alan Templeton Centre for Physical Electronics and Materials SEEIE South Bank University 103 Borough Road, London SE1 0AA TEL 44 171 815 7521 /7571 FAX 44 171 815 7599 email templea-at-sbu.ac.uk
There are a number of standards to look at, and it is possible to order all of them from the various standards committees. One of the most common is the IT8 target, which you can see at http://www.imagequality.com/colmgt.html.
However, there are many calibration charts around for different imaging systems, and the IT8 is not the only one that might be appropriate.
One good place to start looking is the Colour Technology Forum web page at
Another place to look is the Colour Science and Technology Web Page at http://ziggy.derby.ac.uk/web/colour.html.
billo
On Thu, 11 Jun 1998, Barbara Foster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } .....Which brings us to an interesting question regarding color } rendition.... } } } If there were a good standard available, mounted on a traditional 1"x3" } slide, at } } reasonable price ($35-50), would a "color test plate" be of interest to } most people } } who did color video/digital photography? } } } Barbara Foster } } Consortium President } } {bold} {italic} {color} {param} 0000,8080,0000 {/param} Microscopy/Microscopy } Education } } {/color} {/italic} {/bold} 125 Paridon Street Suite 102 } } Springfield, MA 01118 } } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } } Visit our web site: { {http://www.MME-Microscopy.com/education} } } ****************************************************** } } {bold} {italic} {bigger} {bigger} MME: {/bigger} {/bigger} {/italic} {/bold} } America's first national consortium dedicated to } } customized on-site training in all areas of } } microscopy, sample preparation, and image analysis. } } {color} {param} 0000,8080,0000 {/param} Our goal: immediate growth in your productivity! } } {/color} } } } } } } At 05:10 AM 6/11/98 -0700, Bob Lawrence wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Gentlefolk, } } } } } } As usual in all scientific work, the integrity, education, } } } background and reputation of the individual generating the images is the } } } bottom line when it comes to "truth" contained within the images, or any } } } other work one does. The characteristics aren't exclusive, a "good" } } } scientist can still make judgmental and emotional errors, but one has to } } } start somewhere. If the images contain information that is clear and } } } well documented I don't see that it is terribly important to know } } } exactly how they were generated ( except possibly from the point of view } } } of replicating the work). } } } } } } } } } } } } } } } } }
In my previous note about the IT8.7 target, I didn't tell you where you could get one. Sorry. They are sold by most folk who manufacture scanners, etc. I know that Kodak and Agfa sell them.
billo
On Thu, 11 Jun 1998, Barbara Foster wrote:
} } If there were a good standard available, mounted on a traditional 1"x3" } slide, at } } reasonable price ($35-50), would a "color test plate" be of interest to } most people } } who did color video/digital photography? } }
Any idea is good one at this time. However, I forsee two issues that prevents standards to be set, the example here being in making "color test plates"as a slide reference to a digital colorist. 1) control of setting the color microscan standards: the contol of what is sellable does not belong with the microscopist or colorist. In my case the control belongs to the selling agent. That person serves the public buying interest. This may be contrary or compatable to "scientific truth". 2) every scan is different: I have colored insects and bacterias, etc. differently for each scan in order to make a greater selling pool. Accentuating the perspective of the subject can change scan to scan. Renee
........................................................................................ Renee Recker Digital Design Communication Design for Biologists Web, Graphic, and Colored Microscans 16 West 16th St. NYC, NY 10011 212-675-1665 heyrenee-at-renorex.com http://www.renorex.com
Barbara Foster wrote:
} ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------. } .....Which brings us to an interesting question regarding color rendition.... } } If there were a good standard available, mounted on a traditional 1"x3" slide, at } reasonable price ($35-50), would a "color test plate" be of interest to most people } who did color video/digital photography? } } Barbara Foster } Consortium President } Microscopy/Microscopy Education } 125 Paridon Street Suite 102 } Springfield, MA 01118 } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } Visit our web site: {http://www.MME-Microscopy.com/education} } ****************************************************** } MME: America's first national consortium dedicated to } customized on-site training in all areas of } microscopy, sample preparation, and image analysis. } Our goal: immediate growth in your productivity! } } At 05:10 AM 6/11/98 -0700, Bob Lawrence wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Gentlefolk, } } } } As usual in all scientific work, the integrity, education, } } background and reputation of the individual generating the images is the } } bottom line when it comes to "truth" contained within the images, or any } } other work one does. The characteristics aren't exclusive, a "good" } } scientist can still make judgmental and emotional errors, but one has to } } start somewhere. If the images contain information that is clear and } } well documented I don't see that it is terribly important to know } } exactly how they were generated ( except possibly from the point of view } } of replicating the work). } } } } } } } } } }
This is a good analogy, but not really a direct comparison. The mechanisms behind the freezing point depression are different between cooling eutectic alloys and cooling soluble chemical mixtures. In eutectic alloys cooling is controlled by the pooling of pure element regions within the mixture. The smallest pools eg. largest amount of pure metal pooling occurs at the eutectic composition. Then to ask why that condition causes the lowest "freezing" point is another question altogether (best answered by people with big big materials science degrees). In soluable chemical mixtures I don't believe you see any such pooling of the respective organics, so the mechanism for that is different. You must think about each percentage mixture as a bulk chemical. I am sure that there are many organic mixtures that have cooling point spikes as well which is not the case for eutectic alloys. Correct me if I'm wrong.
At 03:26 PM 6/11/98 +-200, you wrote:
} Forgive my ignorance, but if the freezing point of isopentane is 113 K and } that of propane is 84 K, how come a mixture of these freezes at } temperatures below 77 K (boiling LN2)? } Granted that dissolving a solid like salt in a solution leads to freezing point depression. However, I think the freezing point depression in this case might be better compared to a solution of two miscible liquids. I am used to the phenomenon being discussed with application to metals. A solution of tin and lead is liquid below the freezing points of the individual metals. With a "eutectic" mixture you encounter the maximum freezing point depression. I suppose the same principle applies here, even though the compounds are normally liquids (or even gases) at 'normal' conditions.
Dear colleagues, I have a Leaf MicroLumina Scanning Camera which we run on a Windows NT 4.0 System. The camera runs as a Twain plug in device under Adobe Photoshop 4.0. Here is the problem. We start the system up, capture the first image, and then save it to disk. We then go to collect a second image by reselecting the import and selecting the twain device ( MicroLumina), and here there is no response. The program has frozen. One then has to go to the NT Task manager, where you see that Adobe is not responding, and have the system end the Task. Upon restarting the Adobe program a second time, the Lumina runs fine, and no longer it or Photoshop does not hang up. Scitex Corporation is not able to give me any answers to this problem, and has no idea what is causing it. I have specifically asked if this camera was tested on NT 4.0 and With Adobe Photoshop 4.0 and was told that other systems are out there with no problems, but was not given a direct answer to my question. The format we save the file in does not appear to have anything to do with this. The Photoshop program has become hung up, as one cannot even open any other image file in PhotoShop. Has anyone experienced this with their MicroLumina on a Windows NT 4.0 system. It would not surprise me if this is related to Adobe 4.0, as version 4.0 has had some significant changes, with many not for the better. Hope to hear some good news.
Regards,
Joseph Goodhouse Confocal / EM Core Lab Manager Department of Molecular Biology Princeton University jgoodhose-at-molbio.princeton.edu 609-258-5432
Info and Images at http://www.molbio.princeton.edu/confocal/CF-EM-HOME.html
ok, ok, OK, ALREADY. Sorry about the tif posting and any trouble it caused. If you are not going to play or don't need the info- don't look at the file.
Also, in reference to the $75 fee on the recent form, some of the people who have replied have already registered at the cost of $65. That amount ($65) willl be honored and anyone who has already registered at the higher fee will receive a $10 refund.
The story behind the change had to do with the club we were going to play at originally. That club's membership voted to prohibit outside tournament play - - AFTER we had already given the info to MSA for their printing. So, we were stuck with having to find another club on short notice and thus have higher expenses because of it. In any event, we will simply bear the costs and work with the current club.
Call if you have any additional needs or questions.
Thanks, Mark W. Rigler, Ph.D., MAS Inc. 800-421-8451
Hi, does anyone knows where I can get confocal operation training? or is there any workshop for confocal operation? Your help and infomation will be greatly appreciated. Thank you.
Does anyone have experience looking at proliferating cells labelled with BrdU at the EM level? What resin is best, and is DAB/OsO4 localization adequate or are gold/silver probes necessary? Thanks, in advance, for any assistance.
Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219
TO: Nancy Zjaba National Semiconductor South Portland, ME
Soft Imaging System has image analysis software called analySIS. We also have specific modules for grain analysis. If you like more information, please visit our web site http://www.soft-imaging.de or contact us at sales-at-soft-imaging.com or myself and we will be glad to talk to you and send you brochures.
Kuni Tatsumoto kt-at-soft-imaging.com (888) FIND-SIS
This is a good analogy, but not really a direct comparison. The mechanisms behind the freezing point depression are different between cooling eutectic alloys and cooling soluble chemical mixtures. In eutectic alloys cooling is controlled by the pooling of pure element regions within the mixture. The smallest pools eg. largest amount of pure metal pooling occurs at the eutectic composition. Then to ask why that condition causes the lowest "freezing" point is another question altogether. Apparently the heat capacity of that composition is greatest for the given system.
In soluable chemical mixtures I don't believe you see any such pooling of the respective organics, so the mechanism for that is different. You must think about each percentage mixture as a bulk chemical. Of course you get a cooling curve because the heat capacity of neighboring mixtures in a chemical system vary in small amounts at a time. I am sure that there are many organic mixtures that have cooling point spikes as well which is not the case for eutectic alloys. Correct me if I'm wrong.
At 03:26 PM 6/11/98 +-200, you wrote:
} Forgive my ignorance, but if the freezing point of isopentane is 113 K and } that of propane is 84 K, how come a mixture of these freezes at } temperatures below 77 K (boiling LN2)? } Granted that dissolving a solid like salt in a solution leads to freezing point depression. However, I think the freezing point depression in this case might be better compared to a solution of two miscible liquids. I am used to the phenomenon being discussed with application to metals. A solution of tin and lead is liquid below the freezing points of the individual metals. With a "eutectic" mixture you encounter the maximum freezing point depression. I suppose the same principle applies here, even though the compounds are normally liquids (or even gases) at 'normal' conditions.
After a long use of the gold coater, the gold target is thinner and thinner and finally there is a hole showing up. I believe it is the time to replace the gold target.
My question is whether I can temporally cover the hole with a small piece of gold plate so I can still use the existing gold target, before I replace it with a new one. Any suggestions about how to connect two gold plates together (glue them with carbon paste ?). Thanks.
If you are interested in specific video test images, you might look at the Hans Irtel's site at the University of Mannheim, particularly the color CRT page and the Video Test Image page.
Go to http://www.uni-mannheim.de/fakul/psycho/irtel/cvd.html
and take the link to Video Test Images.
You might also look at the CIE site, which deals with standards quite a bit.
Hope this helps.
billo
On Fri, 12 Jun 1998, Thomas A Baginski wrote:
} differently for each system. A color Standard "slide" would be a } terrific start, and would at least allow the same starting point and } settings to be made on hardware.....
We are looking for a TEM tissue processor similar to the histological instruments that are available but one that would allow the automated (or nearly so ) fixation, dehydration, embedding and polymerization into various resins. I believe BAL-TEC used to make a freeze substitution unit that appeared to allow a number of resins and protocols, the FSU 010 but I can't find any sources of information regarding this or similar instruments. Any leads or comments would be appreciated.
I wanted to second John Mansfield's suggestion that those of you who are planning to attend Microscopy and Microanalysis '98 and have not yet made hotel reservation do so as soon as possible. As of our most recent information the Marriott and the Westin still have rooms available in the M&M'98 block. You will need to contact the hotels directly at:
In some cases, we have been told that the hotels report being sold out, although there are M&M rooms available. You may want to check with the M&M contact people:
As a last resort, you can call the M&M Meeting Manager (Bud, Mary Beth, or AnnaMarie) at 708-361-6000 for help.
ALSO PLEASE REMEMBER THAT MONDAY, JUNE 15 IS THE FINAL DEADLINE FOR EARLY REGISTRATION. Registration forms are available on the MSA website or from the Meeting Manager. They need to be faxed to 708-361-6166.
I am not a microscopist, but hope that someone on the list might share some insights regarding sample preparation for wood composite materials. I am trying to prepare thin sections for optical microscopy of waferboard type composites. My need is to obtain sections perpendicular to the wood flake direction. So far, I end up with sawdust. I have tried to impregnate with spur and ultra spur epoxies - to no avail. The costraint I am bound by is that I cannot introduce any moisture or solvent into the matrix which may redistribute its components (adhesive, wax, additives). The typical composite has between 3% and 7% moisture content, an equal amount of isocyanate adhesive, and less than 1% each of wax and other additives. These additives can be water and/or organic solvent soluble.
All suggestions are welcome. I am currently at a loss as to how to proceed.
Regards,
Glenn M. Larkin Graduate Assistant Institute of Wood Research Michigan Technological University Houghton, MI 49931-1295 USA (906) 487-3316
} Does anyone have experience looking at proliferating cells labelled with } BrdU at the EM level? What resin is best, and is DAB/OsO4 localization } adequate or are gold/silver probes necessary? } Thanks, in advance, for any assistance. } } Ronnie Houston } Cytochemistry & Molecular Pathology } Texas Scottish Rite Hospital for Children } 2222 Welborn Street } Dallas, TX 75219
Try Histochemistry 95:491-494, 1991.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Can someone please help Matija with this question?
} Date: Fri, 12 Jun 1998 11:37:17 -0400 (EDT) } From: Matija Maretic {matija-at-gravity.phys.utk.edu} } To: paul.perkes-at-asu.edu } MIME-version: 1.0 } } Dear Mr. Perkes, } } My name is Matija Maretic and I am doing research at University of } Tennessee at Knoxville. I know that Kodak carries a number of } scientific glass plates which can produce high quality positive image, but } I am wondering if they carry a plate which can produce a high quality } black & white negative. } Do you know where I can find this kind of information? } } Thank you, Matija Maretic }
Thanks,
Paul R. Perkes (602) 965-5218 Senior Applications Systems Analyst (602) 965-9004 FAX Center for Solid State Science (602) 952-9583 Wireless Arizona State University paul.perkes-at-asu.edu Box 871704 Tempe, AZ 85287-1704
I suggest you go to the MSA national metting this camming July 12-16. There are a lot of vendors which will be very helpful answering all your questions.
where can an individual purchase stains and chemicals for use under the microscope. Biological supply houses will not sell to me since i am not a company or school.
Thx for info. What is good source for narrowing down its decade of origin?
I have a tremor -- a shaky hand -- so I don't mouse so I don't do WINDOWS or any kind of graphics; sorry. Send me a snailmail address & I'll send you a photo.
Bob & Esther Rowntree 808 Murray Ave San Luis Obispo, CA 93405 (805)549-9107
On Wed, 10 Jun 1998 COURYHOUSE-at-aol.com wrote:
} Gundlach is the mfr! } Good scope! } Wish it was in our collection! } } Take a photo, scan it and share it with all of us! } } } Ed sharpe, archivist smecc }
} Date: Fri, 12 Jun 1998 12:53:05 -0700 } From: Paul R. Perkes {paul.perkes-at-asu.edu} } To: csssano-at-asu.edu } Cc: Matija Maretic {matija-at-gravity.phys.utk.edu} , } Microscopy-at-sparc5.microscopy.com } Subject: Kodak Plate Question } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Can someone please help Matija with this question? } } } Date: Fri, 12 Jun 1998 11:37:17 -0400 (EDT) } } From: Matija Maretic {matija-at-gravity.phys.utk.edu} } } To: paul.perkes-at-asu.edu } } MIME-version: 1.0 } } } } Dear Mr. Perkes, } } } } My name is Matija Maretic and I am doing research at University of } } Tennessee at Knoxville. I know that Kodak carries a number of } } scientific glass plates which can produce high quality positive image, but } } I am wondering if they carry a plate which can produce a high quality } } black & white negative. } } Do you know where I can find this kind of information? } } } } Thank you, Matija Maretic } } } } Thanks, } } } Paul R. Perkes (602) 965-5218 } Senior Applications Systems Analyst (602) 965-9004 FAX } Center for Solid State Science (602) 952-9583 Wireless } Arizona State University paul.perkes-at-asu.edu } Box 871704 } Tempe, AZ 85287-1704 } } Why don't you call Kodak. They have excellent tech reps. The number on their chemical catelog is 800 225-5352; you might have to get another number from them for the film division.
If this doesn't get you what you want, write me with specific info on exactly how you want to use these. There are ways to make reversal images.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We have moved our lab (groan) and the new site is not ideal for our scanning microscope. It looks like we need a vibration isolation table on the column section. I'm looking for recommendations for good vendors.
Many of the facility's users who do negative staining with uranyl acetate end up with an artifact that is most easily described as bubbles. These round areas of no stain are not rimmed and do not appear to be any kind of stuff, but merely absence of stain. Unfortunately, when one is looking for small virus particles and the background is covered with these un-spots, it's horribly distracting.
Can anyone shed some light on this artifact? Before I tear my hair out?
Thanks in advance,
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I remember reading somewhere that we should not wrap contianers of uranyl acetate in aluminum foil. Why is that? Is there a danger of bremstrunghumuhumunukunukuapuaa radiation? (Sorry, it's Friday afternoon and I clearly don't know what I'm talking about.)
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
In response to: where can an individual purchase stains and chemicals for use under the microscope. Biological supply houses will not sell to me since I am not a company or school.
John
There are local science stores that have limited its of slides and chemicals for use with " toy" type microscopes. I would imagine the chemicals would work fine. though.
In Phx. we have one called the 'imaginarium' that sells such things.
Ask Kodak by email (they may be on the web, Kodak have a large excellent site) for the film's processing instructions. I expect that they provide for a reversal procedure (develop, expose, bleach, re-develop, fix). I expect that by using a normal procedure (develop, rinse, fix, wash, dry) these plates will behave as a normal negative film. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} } } } My name is Matija Maretic and I am doing research at University of } } Tennessee at Knoxville. I know that Kodak carries a number of } } scientific glass plates which can produce high quality positive image, but } } I am wondering if they carry a plate which can produce a high quality } } black & white negative. } } Do you know where I can find this kind of information? } } } } Thank you, Matija Maretic } } } } Thanks, } } } Paul R. Perkes (602) 965-5218 } Senior Applications Systems Analyst (602) 965-9004 FAX } Center for Solid State Science (602) 952-9583 Wireless } Arizona State University paul.perkes-at-asu.edu } Box 871704 } Tempe, AZ 85287-1704 } } }
Dear friend, Andrei Chuvilin asked me to send you some references in O/Ag systems. I transfer to you a block of references of one of the last my papers in Catal.Letters. 47(1997)111. What is your interest in this field? because oxygen/silver system is very complicated system. With best wishes, Dr. Andrei Boronin
} In response to: } where can an individual purchase stains and chemicals for use under the } microscope. Biological supply houses will not sell to me since I am not a } company or school. } You're too pessimistic. Companies such as Carolina Biological Supply will sell to anyone. Call 800-334-5551; there are lots of stains listed in their "Science & Math" catalog
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
The Laboratory of Neurobiology, NINDS, NIH (Bethesda, MD) has an opening for an experienced electron microscopy technician. Applicants should be knowledgable concerning the theory, principles, practices and techniques of biological electron microscopy, including operation of transmission EMs and methods for specimen preparation (fixation, embedding, staining, vacuum evaporation, immunolabeling). Competence in cryo-preparation techniques (rapid freezing, cryo-ultramicrotomy, freeze-fracture, freeze-substitution) is especially desirable. Experience with specialized fields of elemental mapping and analysis (x-ray microanalysis, electron energy loss spectroscopy, STEM, and energy-filtering TEM) would be an additional advantage. The NIH offers competitive salary and benefits packages; US citizenship required. For more information contact Brian Andrews at "sba-at-helix.nih.gov".
The phenomenon that you describe has been called champaigning. It is thought to be caused by a number of factors (none of which have been tested and proven to be the cause, however). This includes: hydrophobicity of the substrate - usually Formvar/carbon, improper spreading of the suspension, inadequate drying of the specimen on the grid, inclusion of substances - like sucrose or salts - that boil in the beam, actual particles such as ribosomes or glycogen or viral capsomeres.
Here are some things to try: - a different stain (one of the tungstates) to rule out "real" particles versus bubbles, - glow-discharge treating the grids to eliminate hydrophobicity, - inclusion of a wetting agent such as bovine serum albumin to help spreading of the specimen, - allow the grid to dry for at least 20 minutes (preferably at 45-60 C), - do not focus the beam on the specimens (causes boiling), - make sure you have a thin spread (not clumped), - offer a sacrifice to Pele or Vulcan.
Cheers, John
########################### Dr. John Bozzola, Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901 Phone: 618-453-3730 Fax: 618-453-2665 ###########################
UrAc is thought to be sensitive to light - especially UV type of light found in fluorescent light and sunlight. Causes breakdown of the stain and possible precipitation. Very little radioactivity is present - but some is there, so be careful.
JB
########################### Dr. John Bozzola, Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901 Phone: 618-453-3730 Fax: 618-453-2665 ###########################
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Long Liang wrote: ================================================ After a long use of the gold coater, the gold target is thinner and thinner and finally there is a hole showing up. I believe it is the time to replace the gold target.
My question is whether I can temporally cover the hole with a small piece of gold plate so I can still use the existing gold target, before I replace it with a new one. Any suggestions about how to connect two gold plates together (glue them with carbon paste ?). Thanks. ================================================= This is a novel suggestion. It might even work. However, there should be high conductivity between the "patch" and the rest of the cathode, and for that, we would use the same silver filled epoxy that we recommend when the cathode has to be glued onto the "head". Our prediction is that the carbon paste probably would not have sufficient conductivity. If silver paste was used, we would think that the silver would start to sputter away. When you do your "glue" treatment, make sure you put some weights onto the assembly so that you end up with a very uniform layer of the silver filled epoxy.
Disclosure: SPI offers all of the above discussed items as well as new "replacement" cathodes.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
This is a request for experienced users of Image Analysis software. I would appreciate if some of you are able to spare me some information on the types of software avaliable.
I have been using OPTIMAS at present, and I am interested in other programs which may be easier to use, and/or offer more capabilities. The ability to do FFT would be good.
I know also of "analySIS" and "SigmaScanPro" from thier web pages, but would also like to know some opinions of these from people who have tried the software.
I am also keen to find out prices of such software if they are availiable. I am looking at intermetallic particles in Aluminium Alloys, and currently using B/W backscatter images from the SEM.
We have made excellent experience with a three-axis system sold by
Integrated Dynamics Engineering 84 Cummings park Woburn, MA 01801 tel: 617-938-5120 fax: 617 938 5122 100125.1130-at-compuserve.com (address already mentioned)
The location of our FE-SEM (JEOL 6300F) is far from being ideal. There are a number of large machinery, elavators, printing machines etc next door.
Best regards, Dr. Peter Reynders +-----------------------------------------------------------------+ | Merck KGaA - Pigments & Cosmetics Division | | Bldg M 18, 64271 Darmstadt / Germany | +-----------------------------------------------------------------+ | e-mail: reynders-at-merck.de (work) reynders-at-compuserve.com (home) | | homepage: http://ourworld.compuserve.com/homepages/reynders/ | +-----------------------------------------------------------------+
Rozeveld, Steve (SJ) wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Another possibility: } } Integrated Dynamics Engineering } 84 Cummings park } Woburn, MA 01801 } tel: 617-938-5120 } fax: 617 938 5122 } 100125.1130-at-compuserve.com } } } Steve Rozeveld } } } } Dow Chemical Co. } } 1897 Bld. Door E-43 } } Midland, MI 48667 } } sjrozeveld-at-dow.com } } % (517) 636-5167 } } Fax: (517) 638-6443 } } } ---------- } } From: Ron Doole[SMTP:ron.doole-at-materials.oxford.ac.uk] } } Sent: Friday, June 12, 1998 2:08 AM } } To: Microscopy Community } } Subject: Re: SEM - problems with magnetic interference } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } On Thu, 11 Jun 1998, Alan Templeton wrote: } } Hi Alan, } } } } We have have been very interested in this problem as well. We have } } had trials of, and heard other good reports of, the field cancelling } } system sold by Oxford Instruments. Contact them at: } } Oxford Instruments, } } Research Instruments, } } Tubney Wood, } } Abingdon, } } Oxon. } } OX13 5QX. } } 01865-393200. Ask for Judith Brock. } } e-mail:- judith.brock-at-oxinst.co.uk } } } } Unfortunately I have no connections or financial interests. } } } } Good luck, } } Ron } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Does anybody know of a manufacturer or supplier of magnetic field } } } containment loops. We are having problems with our Hitachi S-800FE } } } SEM - getting possible structurally transmitted magnetic interference } } } from lab two floors up. Our SEM is located in a basement with the } } } structural columns (steel reinforced concrete) directly above and to the } } } side of the SEM. Interference manifests itself in the form of CRT } } } screen "wobble" of set frequency especially noticable at high mag } } } (} x3K). Equipment in other lab determined to be the source of the } } } problem is a Microwave plasma reactor (interference occurs when } } } heater is on) and an Electro-cyclotron resonance (ECR) instrument. } } } Need urgent help.Cheers } } } Dr Alan Templeton } } } Centre for Physical Electronics and Materials } } } SEEIE } } } South Bank University } } } 103 Borough Road, London } } } SE1 0AA } } } TEL 44 171 815 7521 /7571 } } } FAX 44 171 815 7599 } } } email templea-at-sbu.ac.uk } } } } } } } =========================================================================== } } Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk } } Department of Materials, phone +44 (0) 1865 273701 } } University of Oxford, fax +44 (0) 1865 283333 } } Parks Road. } } Oxford. OX1 3PH. UK. } } ============================================================================ } }
We have a Phillips EM 420 TEM. It is getting on in years, but is still a lovely machine. We have a slight problem, though. The op-amp on the board for the auto-exposure system seems to blow after only two to three weeks use. We are not sure what the problem is and have been told by the local Phillips guys that ours is the only machine that does this. Short of trying to get a new/replacement board, are there any suggestions/plans/calls of sympathy out there? Any help will be appreciated. Thanks!
A colleague of mine is preparing x-sectional TEM specimens of CdZnTe/CdHgTe epilayers. We are using a new Gatan PIPS ion mill. The problem is that some specimens come out very nicely, but others are completely polycrystalline. The specimens are all prepared under the same/closely similar conditions, but we are unable to make reproducible TEM samples. I was therefore wondering whether there are any research facilities out there with a FIB machine that might have some time available on the machine to make some samples for us. Please advise as to the availability of the machine, and rates which apply for use, etc.
The Lynx Tissue Processor (now sold by Leica) is an instrument I have used for nearly 13 years-both here and at my previous employer. Operationally, the Lynx (and the RMC processor-a descendent of the old LKB processor) provides 20 stations that carry reagents up to pure resin. Both are carousel designs (with some differences) that have vials for reagents. I have used these for osmium through pure resin. A number of programs can be entered into memory, and the processing schedule tailored for the type of resin and the different dehydrants and en bloc staining protocols. It is theoretically possible to use the machine for immunocytochemistry or enzyme cytochemistry protocols, but I have not utilized them for those procedures.
The original instrument (purchased from Lynx before Leica took it over) was reliable and operated smoothly for the 4 years I was in that lab. I cannot give as ringing an endorsement to the unit we purchased here in 1990. Over the first 4 or 5 years, although there was low demand for use, we maintained the unit and did routine test runs. However, it has been necessary to send the unit back to Leica more times than I can (or care to) remember. It has run quite consistently for the last six months, during which time I have needed to process large numbers of samples daily for several weeks (I'd really rather not think about it). Hopefully, after nearly 8 years, the bugs have been worked out.
I have no experience with the RMC unit, and have been unable to get them to provide a demo unit for actual lab testing. That is frustrating. I wish you the best in your quest.
Roger Moretz, Ph.D. Dept of Toxicology
-----Original Message----- From: John N. Wright [SMTP:johnw-at-uts.cc.utexas.edu] Sent: Friday, June 12, 1998 2:40 PM To: Microscopy Community Subject: TEM Tissue Processors
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We are looking for a TEM tissue processor similar to the histological instruments that are available but one that would allow the automated (or nearly so ) fixation, dehydration, embedding and polymerization into various resins. I believe BAL-TEC used to make a freeze substitution unit that appeared to allow a number of resins and protocols, the FSU 010 but I can't find any sources of information regarding this or similar instruments. Any leads or comments would be appreciated.
Good timing. I have in front of me the latest copy of "The Microscope Book" (The Cambrex Group Mail Center, Suite 2, 112 Washington St, Marblehead, MA 01945-3554). There are three companies in this issue:
Fabreeka International, Inc. 1-800-FABREKA www.fabreeka.com {http://www.fabreeka.com}
Herzan 30251 Golden Lantern Laguna Niguel, CA 92677 949-363-2905 73071.3720-at-compuserve.com {mailto:73071.3720-at-compuserve.com}
TMC is the only one I know (personally) that makes anti-vibration units for EMs, but the others might be worth checking out.
Roger Moretz, Ph.D. Dept of Toxicology
-----Original Message----- From: Ron L'Herault [SMTP:lherault-at-bu.edu] Sent: Friday, June 12, 1998 8:02 PM To: microscopy-at-sparc5.microscopy.com Subject: vibration isolation tables
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We have moved our lab (groan) and the new site is not ideal for our scanning microscope. It looks like we need a vibration isolation table on the column section. I'm looking for recommendations for good vendors.
The first thing I would do is to check out the power supply for spikes and check grounding. Since the problem seems intermittant, it may prove difficult to "catch" it misbehaving. Another possibility may be inductive "kick-baqck". If there are any relays or solenoids associated with the circuit, a "snubber" diode across the coil may have gone open. That would permit a high voltqge spike to superimpose on the power supply.
In any case, you might try protective zener diodes. If the opamp uses +- voltage, one would be needed for each polarity feed. Choose a Zdiode with a rating a volt or two over the P.S. voltage. Connect from power feed to ground AT the opamp. Be sure to observe correct polarity.... If the added capacitance and the tiny leakage current will not be a problem, the same can be done for the inputs and output. If the I/O is bipolar, then two Zdiodes, wired in series - cathode to cathode should be used. The voltage rating of each these diodes should be slightly greated than the expected excursion of the I/O line it is protecting
Good Luck! Woody ------------------------------------------------ Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
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Hallo All
We have a Phillips EM 420 TEM. It is getting on in years, but is still a lovely machine. We have a slight problem, though. The op-amp on the board for the auto-exposure system seems to blow after only two to three weeks use. We are not sure what the problem is and have been told by the local Phillips guys that ours is the only machine that does this. Short of trying to get a new/replacement board, are there any suggestions/plans/calls of sympathy out there? Any help will be appreciated. Thanks!
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Hi Everyone, A long time ago (about 13 years) we used to use 3 1/4"X4" glass negative plates made by Kodak their catalog number was 185-5881. Maybe you could contact Kodak and inquire if they still make them. I hope this information helped. Jane Glamp PPG Inds.
Can someone please help Matija with this question?
} Date: Fri, 12 Jun 1998 11:37:17 -0400 (EDT) } From: Matija Maretic {matija-at-gravity.phys.utk.edu} } To: paul.perkes-at-asu.edu } MIME-version: 1.0 } } Dear Mr. Perkes, } } My name is Matija Maretic and I am doing research at University of } Tennessee at Knoxville. I know that Kodak carries a number of } scientific glass plates which can produce high quality positive image, but } I am wondering if they carry a plate which can produce a high quality } black & white negative. } Do you know where I can find this kind of information? } } Thank you, Matija Maretic }
Thanks,
Paul R. Perkes (602) 965-5218 Senior Applications Systems Analyst (602) 965-9004 FAX Center for Solid State Science (602) 952-9583 Wireless Arizona State University paul.perkes-at-asu.edu Box 871704 Tempe, AZ 85287-1704
You should try the small angle cleavage technique for these samples. No ion milling at all. You just need a delicate touch. Look up John McCaffrey and my paper in the TEM sample prep IV book from MRS, vol 480, 1997. This is a detailed, pictorial outline of how to do it. Send me your address and I'll send you a reprint. We are also going to have are poster at M&M98 on the technique. South Bay Technology has a kit that would cost you about the same as a day on an FIB.
What is the substrate? If it is a II-VI, and you use the technique, I would recommend a thickness of about 100-110 um for the final thickness. If you have problems, instead of the 15 degrees, you might want to go a little higher, say 18 degrees.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
A colleague of mine is preparing x-sectional TEM specimens of CdZnTe/CdHgTe epilayers. We are using a new Gatan PIPS ion mill. The problem is that some specimens come out very nicely, but others are completely polycrystalline. The specimens are all prepared under the same/closely similar conditions, but we are unable to make reproducible TEM samples. I was therefore wondering whether there are any research facilities out there with a FIB machine that might have some time available on the machine to make some samples for us. Please advise as to the availability of the machine, and rates which apply for use, etc.
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} } The Lynx Tissue Processor (now sold by Leica) is an instrument I have } used for nearly 13 years-both here and at my previous employer. } Operationally, the Lynx (and the RMC processor-a descendent of the old } LKB processor) provides 20 stations that carry reagents up to pure } resin. Both are carousel designs (with some differences) that have } vials for reagents. I have used these for osmium through pure resin. A } number of programs can be entered into memory, and the processing } schedule tailored for the type of resin and the different dehydrants and } en bloc staining protocols. It is theoretically possible to use the } machine for immunocytochemistry or enzyme cytochemistry protocols, but I } have not utilized them for those procedures. } } The original instrument (purchased from Lynx before Leica took it over) } was reliable and operated smoothly for the 4 years I was in that lab. I } cannot give as ringing an endorsement to the unit we purchased here in } 1990. Over the first 4 or 5 years, although there was low demand for } use, we maintained the unit and did routine test runs. However, it has } been necessary to send the unit back to Leica more times than I can (or } care to) remember. It has run quite consistently for the last six } months, during which time I have needed to process large numbers of } samples daily for several weeks (I'd really rather not think about it). } Hopefully, after nearly 8 years, the bugs have been worked out. } } I have no experience with the RMC unit, and have been unable to get them } to provide a demo unit for actual lab testing. That is frustrating. I } wish you the best in your quest. } } Roger Moretz, Ph.D. } Dept of Toxicology } } } -----Original Message----- } From: John N. Wright [SMTP:johnw-at-uts.cc.utexas.edu] } Sent: Friday, June 12, 1998 2:40 PM } To: Microscopy Community } Subject: TEM Tissue Processors } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } We are looking for a TEM tissue processor similar to the } histological } instruments that are available but one that would allow the } automated (or } nearly so ) fixation, dehydration, embedding and polymerization } into } various resins. I believe BAL-TEC used to make a freeze } substitution } unit that appeared to allow a number of resins and protocols, } the FSU 010 } but I can't find any sources of information regarding this or } similar } instruments. Any leads or comments would be appreciated. } } Thanks in advance, } John N. Wright }
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
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Dear fellow microscopists and friends, At San Joaquin Delta College, Microscopy Technology Center, we are a = 2 yr microscopy training program, as many of you know. Recently we = posted a position for an Adjunct for Teaching Fall semester 1998 while I = take a sabbatical. Unfortunately the person that signed the contract = for it, decided Friday that he no longer wants to do it. This leaves us = in a real bind. If any of you have ANY ideas for getting these four courses taught = this fall (the position info is listed below with the courses) in = Stockton, CA, please e mail me directly. If you want to apply, contact = Human Resources directly at 209/954-5056 for an application. Perhaps you know someone who is just finishing a post doc and might = like it for the semester, or someone who could take a leave for a = semester from industry or educational institution. OR perhaps you know someone who could teach one of the courses and = is nearby. We are open to any kind of suggestion at this point. The job = offers full benefits and a good salary for the semester. The courses would not require development of new courses as these = courses have been fully developed. Course materials are available to aid = in the instruction. My sabbatical is locked in for Fall as I must beat the bushes to make = $3 million for our new building to match the state's funding of $4.5 = million. Any ideas are welcome. If you could teach the courses, you can get an = application at Human Resources whose phone number is listed below. Any help is appreciated. If this note sounds a bit desperate, IT IS. = Time is of the essence.
Thanks Judy Murphy
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
URGENT: ADJUNCT FACULTY NEEDED FALL Semester 1998 (Aug. 12 - Dec 18, 1998)
A San Joaquin Delta College Faculty member of the Microscopy Technology = Center is planning a Sabbatical Leave for the Fall '98 semester. San = Joaquin Delta Community College is currently searching for an EM = Instructor Adjunct / Temporary One Semester Contract for Fall 98 (Aug. 12 = - Dec 18, 1998).
MINIMUM QUALIFICATIONS: Bachelor's Degree plus two years of directly related experience OR an = Associate Degree plus six years of directly related experience OR a valid = credential.
DESIRABLE QUALIFICATIONS: Master's or PhD Degree in a Biological = Science; Experience in teaching / practice of Electron Microscopy
COURSES TO BE TAUGHT:
Introductory Techniques for Transmission Electron Microscopy (EM21) This is a lecture/lab course which includes beginning Transmission = Electron Microscopy dealing with the alignment and operation of the TEM, = vacuum techniques, photographic techniques, as well as the preparation of = particles and replicas for viewing in the TEM. Includes individual = training in the use of the TEM, preparation techniques, and written and = oral reports. (Lec - 2 hrs; Lab - 3 hrs/wk)
Biological Ultrastructure (EM28) Course contents include specific information about the fine structure and = function of cells and tissue at the ultrastructure level. Videos, slides = and micrograph examination will be correlated with the lectures so that = students will learn to recognize the fine structure of cells and tissues = in relationship to their function. (Lec-2 hrs/wk)
Advanced Techniques in Biological Electron Microscopy (EM37) Course contents include lecture and laboratory which covers advanced = techniques for biological specimen preparation in TEM including an = advanced research project.( Lec - 1 hr; Lab - 6 hr/wks)
** Current Microscopies: Optics, Theory and Application (EM30) Course contents include information related to the physical laws and = applications of the various types of current microscopies e.g. = TEM,SEM,FIB, AFM, and confocal microscopy, as well as other current = topics e.g. asbestos analysis, lab design, etc. (Lec - 2 hrs; Lab - 3 = hrs/wk) ** This course is negotiable. A portion of this assignment would be = paid as an additional overload.
The above teaching load would not require development of new courses as = these courses have been fully developed. Course materials are available = to aid in the instruction. TERMS OF EMPLOYMENT: Full semester, Non-tenured track position with Full = Benefits (Medical, Dental, & Vision). SALARY for Fall 98 semester: BA/BS w/ 2 yrs experience or AA w/ 6 yrs experience$17,498 - 25,333 for Fall 98 sem
MA/MS$19, 219 - 28,137 for Fall 98 sem
PhD$ 22,528 - 31,739 for Fall 98 sem
APPLICATION: Contact Human Resources at 209/954-5056. Human Resources San Joaquin Delta College Admin. Bldg. Rm 202 5151 Pacific Ave Stockton, CA 95207
DEADLINE: Please send your cover letter of application, an application = form for employment, and copies of all transcripts as soon as possible. = The Screening Committee plans to begin the review of applications on June = 29, 1998.
Additional SJDC Microscopy Technology Program Information available at = http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html/
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Dan Caruso asked:
{Are there any advantages to using cacodylate buffers in fixatives as {opposed to phosphate buffers? Are there any disadvantages in addition to {the toxicity of cacodylate buffers?Specifically, I am working with human {corneas. Thanks for your replies.
One of the reported advantages of cacodylate buffers - Andrey Glauert dixit - is that they do not support microbial growth, e.g. fungi. This is almost true. Fungi can actually grow in glutaraldehyde fixed tissue 'eating up' the fixed protein, and traveling throughout less dense spaces, sometimes giving images suitable to illustrate science-fiction 'infections'. We reported this some years ago in
Growth of fungi in cacodilate buffer - J.F. Moura Nunes, A.P. Aguas e J.O. Soares - Stain Technology, 55: 191-192, 1980
Best wishes
J.F.Moura Nunes Lab. Electron Microscopy Portuguese Cancer Institute 1093 Lisbon Codex Portugal
I teach two upper level classes Plant Anatomy & Structure and Function of Algae and Land Plants. I am constructing web pages for them. These can be seen from my home page www.botany.hawaii.edu/faculty/webb/default.htm
I need EM images, especially for plant and algal cell ultrastructure. All algae are welcome. However, SEM images would also be welcome. I would like to post these on our server so that they could be accessed via the Internet. Authors would be credited. I have my own digitizer. Thus, you could send me a photographic slide or negative (color OR black and white) as well as Prints!!!! Otherwise, I could send you a ZIP Disk or Floppy Disks which you could use to transfer files. I will pay postage!!! Finally, I could download them from a web site. Feel free to use any of my images. I am working on Plant Anatomy this summer and that site will be expanding rapidly during July. Finally, if you know of any good websites for the above, please let me know.
In response to the recent discussions concerning victawet we'd like to make the following case:
Ladd has manufactured Vacuum Evaporators for the EM field for over 30 years. Since each one has to be factory tested upon completion it's important that we are able to make them spotless again prior to shipping After trying to accomplish this for years we settled on the following procedure:
1. Evaporate victawet from a tungsten basket which coats the bell jar
2. Test Evaporator
3. Using a soap solution and lint free cloths we then wipe down the internal parts.
This has worked well for us for years.
SUBSTRATES
We periodically do special substrates for some of our customers who insist they be as contamination free as humanly possible. To accomplish this we use victawet prior to the substrate run. This assures the elimination of any possible residual oil.
John Arnott LADD RESEARCH 13 Dorset Lane Williston, VT 05495
***************************** DISCLAIMER **************************** Ladd Research sells victawet, vacuum evaporators, bell jars, substrates and other products mentioned in this e-mail
I am about to begin a phase of a research project involving microtome sectioning of flowers and fruits for developmental studies. I am interested in finding any educational cd's, vcr's, or other resources on the process of rotary microtome sectioning. I have several books and "human" resources for guidance but it would be helpful if there are any other visual or lab teaching aids out there as well.
Thanks in advance for any leads or ideas.
please respond to: crow-at-aloha.net
_______________________________________________________________ Melany H. Chapin
To Reply Please Remove Spam-Proof "XXX" from return address.
ADDRESS: crow-at-aloha.net
After dark, all cats are leopards. (Zuni proverb) We will be known by the tracks we leave. (Dakota) When the legends die, the dreams end; there is no more greatness (Shawnee) Every animal knows more than you do. (Nez Perce) ________________________________________________________________
I don't have the schematics of that particular instrument handy, but here's a simple tip. If there are any transistors being driven by this opamp, replace them. Often in repairing a problem, a failed component will be replaced and the circuit will operate, but related components in the circuit may have been stressed to the point where they make excessive demands on the replaced component.
Another situation arises where a component may get marginal in operation, such as a transistor suffering from thermal runaway. That component may not fail but may cause others to fail. Replacing a component earlier in the circuit may correct the apparent problem, but not the cause - the replaced part will soon require replacement again.
} Hallo All } } We have a Phillips EM 420 TEM. It is getting on in years, but is } still a lovely machine. We have a slight problem, though. The } op-amp on the board for the auto-exposure system seems to blow after } only two to three weeks use. We are not sure what the problem is } and have been told by the local Phillips guys that ours is the only } machine that does this. Short of trying to get a new/replacement } board, are there any suggestions/plans/calls of sympathy out there? } Any help will be appreciated. Thanks! } } Regards } Alistair Douglas Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Many thanks to everyone who replied to my query about non-bubbles with negative staining. The replies confirm what I'd always suspected lo these more than 20 years at EM - it's not just me, no one knows what's going on with some of this stuff. It's all witchcraft.
ohn Bozzola has called the effect Champaigning, which is a lovely description. Many people sent me their pet ideas about what causes the effect, and what to do about it. I can debunk most of them, usually in the course of one staining session. Sigh.
I get it with UrAc, PTA, and uranyl formate. I don't carbon coat my grids, so hydrophobic carbon is not the culprit. There was no glycerol in the sample. Wetting agents have not helped. I can get good spreads, and last week one of the grids was gorgeous, while the other 19 spread well but had bubbles. Happens with all kinds of samples.
The main difference with the one that worked was that it dried longer while I babbled on atthe people I was training. I will pursue this line of investigation.
Sacrifices to Pele generally help, but already the microscope room is full of ti leaves. (Even our engineers get into this.)
As for my other uranyl acetate question- I know UrAc is light sensitive, and so I have always kept it in a dark bottle in the fridge where presumably the light is out when the door is closed. I can't confirm this. Anyway, for the stuff left out in room light, I had always wrapped the container with aluminum foil. I keep stain in 10 cc syringes with millepore filters on them at room temp so they are always ready for staining. I recently read that foil was a bad idea, but with no explanation. Phil Oshel offers one-
"It's worse than you think. UAc is afraid of the dark. When wrapped in foil, it starts shaking with fear, and the nuclei go into resonance, lowering the nuclear-shell transition energies. More neutrons are released, leading to a chain-reaction.
This is know as phobiafusion, and was discovered by a group of unemployeed physicists and molecular biologists in San Fransico who had set up as macromolecular psychotherapists."
It made me laugh.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Does anyone have a fax or email address for a supplier of electronic components who's likely to be able to supply me with 6 or so Burgess microswitches type V4T7-GP ? Local agents have a minumum order of 50!
cheers
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I recommend Newport Micro-Controle. Sorry, I don't know their address in the US since I dealt with their Belgian representative. They custom designed a table for my LEO/Zeiss EM912 which works perfectly. I should also point out the excellent contact and customer service I received.
Usual disclaimer: no personal interest in this company, I'm just a very satisfied customer.
Regards, Michel
**************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
Hi, I have a question on the older NIM bin Ln2 sensor / bias protection modules Tracor Northern used. Module 1236. Does someone know what device they used as the sensor that went in the Ln2. Was it a diode or a thermistor of type ? We have the module that we would like to use on another system, but have no reference to the sensor device.
Thanks Luc Harmsen Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
} } To: {FreidaC-at-aol.com} } From:tflore-at-lsumc.edu (Teresa Flores) } Subject:Re: Spanish unite with NSH } Cc:histonet-at-pathology.swmed.edu } } Frieda, thanks for all your suggestions and your genuine concern. } I thought that you knew that there was is a Sociedad de Latinoamericana de } Histotecnologia (SOLAHT), of which, I am the President and Rosemary } Velasquez is the Secretary-Treasurer. SOLAHT was founded in 1995 in Santa } Cruz, Bolivia. There are approximately over 300 members and over 100 that } I have not had an opportunity to imput or respond to (because of personal } commitments and the tremendous amount of work I have right now). These } members range from Mexico, Guatemala, Panama, Costa Rica, San Salvador, } Nicaragua, Honduras, Bolivia, Ecuador, Paraguary, Argentina, Chile (I } might have left out a country or two). I did not realize how time } consumeing it would be to keep up with the SOLAHT and sometimes I am } overwhelmed. } I believe I will send out a communication to the SOLAHT with information } on becoming an NSH member. I would need NSH International yearly dues and } what they would include. Also, if SOLAHT members want to become NSH } members and not receive the Histology journal, is there a lesser NSH fee? } Or if SOLAHT members just want the Histotechnology journal, is there a } lesser NSH fee? } Frieda, please give me all the particulars and options so that I may relay } the information to SOLAHT members. } In the communication to SOLAHT I will be requesting e-mail addresses to } try and begin a listserver. } Thanks again for all your help. Teresa. } ,} Theresa: } } } } There is no provision for corresponding members in NSH. All members outside } } of the US and Canada are International members and they pay the same dues and } } receive the same benefits as the regular active members. } } } } Hard Tissues do not have an extra day. There are special workshops and } } seminars especially slanted toward the VIR or hard-tissue group, but some are } } attended by clinical members. } } } } We would have to have a guarantee of a certain number of Spanish-speaking } } attendees in order to make special concessions. Rather than an extra day, } } translators might be a better choice. However this would probably cost a lot } } more and an extra charge would have to be made. I do not know how we could } } guarantee attendees either. } } } } If there is as much interest as you say, has there been any thought given to } } South and Central American and Mexico forming a Spanish-speaking Society? } } } } I know I am not being much help - but I do not know what to suggest. } } } } Freida }
To all MSA/MAS '98 meeting attendees (esp. SEMS, AREMS and AMS members), If you don't want to see Centennial Olympic Park or be sippin' mint juleps on the veranda we could use your help staffing the Information and Hospitality Booth. Cynthia Goldsmith and I are coordinating the Booth. If you have a few hours to spare please let me know. No prior experience is necessary and I guarantee your pay stub will have a lot of zeros in it :-).
Thanks for considering this matter. See you in Atlanta, Beth Richardson
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
The Biological Sciences meeting of MMMS will be held on Wednesday, July= 1 at the Blood Research Institute in Milwaukee, Wisconsin. Registration and=
Poster Set-up begin at 10:00 a.m.; the first scientific presentation st= arts at 10:30 a.m.
Registration is free to current members, and attendees may join the soc= iety at the meeting. Dues are $10 for regular members and $5 for students. = No pre-registration is required to attend the scientific sessions; however= , if you wish to have lunch with us, we DO need a headcount and menu selecti= ons by June 26. YOU MUST RETURN YOUR MENU SELECTIONS TO US, IF YOU WANT TO PARTICIPATE IN LUNCH!
The menu, complete agenda, and travel instructions will be sent to curr= ent MMMS members this week. For anyone else interested in attending, pleas= e contact Linda LaRonge-Snow at linda.snow.nmi-at-abbott.com OR telephone (847) 937-5251 OR fax (847) 93= 8-5027 with a fax number or mailing address (fax preferred).
Poster space is available for anyone wanting to show off their work. Available space for each poster is 3 X 5 feet. We'd like to have a rou= gh estimate of the number of posters, so please contact Jim DiOrio at diorio-at-baxter.com if you intend to display a poster. Last minute entr= ies, however, will be accepted!
The meeting agenda is as follows:
10:00 - Registration and Poster Set-up
10:30 - Using Correlative Microscopy in Biological Problem Solving; Ral= ph Albrecht, Ph.D., Univ. WI - Madison
11:15 - Localization of C-Terminal Gamma Chain Regions in Gold-Cadaveri= ne Labeled Fibrinogen by STEM; Michael Mosesson, M.D., Univ. WI Medical = School
Noon - Lunch and Poster Session; Medical College of Wisconsin
1:00 - Digital Manipulation of Acquired Images: "What Is Possible vs. W= hat is Ethical"; John Kinnamon, Ph.D., Univ. Denver; MSA/LAS sponsored speake= r
1:45 - Roses and Feathers; Joseph Harb, Ph.D., Diagnostic EM, Med. Coll= ege of Wisconsin
2:30 - Microscopic Characterization of T-DNA Tagged Male-Sterile Arabid= opsis thalianaLines; Heather Owen, Ph.D., Univ. WI - Milwaukee
3:15 - Refreshments
3:30 - Digital Imaging Workshop - John Kinnamon, Ph.D.
For further information, you can contact me at jane.a.fagerland-at-abbott.= com. Hope to see you in Milwaukee on July 1st!
Dear Tina, } } I remember reading somewhere that we should not wrap contianers of uranyl } acetate in aluminum foil. Why is that? Is there a danger of } bremstrunghumuhumunukunukuapuaa radiation? (Sorry, it's Friday afternoon } and I clearly don't know what I'm talking about.) } I am not sure why one wouldn't wrap UAc containers in Al foil, but it has nothing to do with radiation. The only kind of radiation which will penetrate the bottle to interact with the foil would be gammas from some of the daughters of the U. If one had a very thin-walled container and a very energetic beta, one might have to worry about brehmsstrahlung radiation (It's Tuesday morning & I'm still awake), but since its production goes as a power of Z, Al is one of the better metals for avoiding this problem. Be is the best mechanically sound metal for avoiding brehmsstrahlung, but has other problems. Plastic is another good material for avoiding it. Yours, Bill Tivol
Phobia*fission*! Sorry. I was so worried about the effects of a small nucleopsychotic reaction in Tina's 'fridge that I mixed up my fused and split personalities.
Phil
} As for my other uranyl acetate question- } I know UrAc is light sensitive, and so I have always kept it in a dark } bottle in the fridge where presumably the light is out when the door is } closed. I can't confirm this. Anyway, for the stuff left out in room } light, I had always wrapped the container with aluminum foil. I keep } stain in 10 cc syringes with millepore filters on them at room temp so } they are always ready for staining. I recently read that foil was a bad } idea, but with no explanation. Phil Oshel offers one- } } "It's worse than you think. UAc is afraid of the dark. When wrapped in } foil, it starts shaking with fear, and the nuclei go into resonance, } lowering the nuclear-shell transition energies. More neutrons are } released, leading to a chain-reaction. } } This is know as phobiafusion, and was discovered by a group of unemployeed } physicists and molecular biologists in San Fransico who had set up as } macromolecular psychotherapists." } } It made me laugh. } } Aloha, } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 5037 Station A Champaign, IL 61825-5037 (217) 355-1143 oshel-at-shout.net or poshel-at-hotmail.com ***** looking for a job *****
The real reason for not using aluminum foil is that the acetate ion will decompose the aluminum (if liquid dribbles down the side of the container --- gasp --- and onto the foil). This generates a crumbly mess that is contaminated with Ur to boot. If one carefully pipettes the UrAc, the contact is avoided. We have used aluminum to cover our UrAc for several decades and only once did the Al decompose (in out student laboratory).
} } I remember reading somewhere that we should not wrap contianers of uranyl } } acetate in aluminum foil. Why is that? Is there a danger of } } bremstrunghumuhumunukunukuapuaa radiation? (Sorry, it's Friday afternoon } } and I clearly don't know what I'm talking about.) } } } I am not sure why one wouldn't wrap UAc containers in Al foil, but } it has nothing to do with radiation. The only kind of radiation which will } penetrate the bottle to interact with the foil would be gammas from some } of the daughters of the U. If one had a very thin-walled container and a } very energetic beta, one might have to worry about brehmsstrahlung radiation } (It's Tuesday morning & I'm still awake), but since its production goes } as a power of Z, Al is one of the better metals for avoiding this problem. } Be is the best mechanically sound metal for avoiding brehmsstrahlung, but } has other problems. Plastic is another good material for avoiding it.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
The TN 1236 Bias Protection Module was actually designed and built by Tracor X-ray which is now Spectrace Instruments, 1275 Hammerwood Avenue, Sunnyvale, CA 94089 USA. phone 408-744-1414 www.spectrace.com Wayne Watson or Bill Drummond could help you on this. I believe the sensor was a thermistor. They may still use the same sensor in their XRF spectrometers with LN detectors.
Ronald Vane XEI Scientific
Luc Harmsen wrote: } } Hi, } I have a question on the older NIM bin Ln2 sensor / bias protection modules Tracor Northern used. Module 1236. } Does someone know what device they used as the sensor that went in the Ln2. Was it a diode or a thermistor of type ? } We have the module that we would like to use on another system, but have no reference to the sensor device. } } Thanks } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za
I have been asked to recommend a stereo light microscope and camera combination for use in a classroom situation. I turn to you, the experts, for suggestions, advice, and ideas.
The microscope will be used in a plankton class to record images of 'things'. The pictures will be used to share with other students in the class so everyone can see the 'things' everyone else collected. The pictures might also be used in student reports and possibly made into 2x2 slides for teaching. Images of the highest quality are not required, but they should be reasonably good.
I am not familiar with the latest in stereo microscopes so some pointers here would be helpful. Are there any models that stand out as particularly good candidates for this type of application?
For this application I suggested some sort of digital camera and printer. Could a Pixera system fulfill the needs of this user? How about an inexpensive color printer to have nearby?
At the moment, there is no budget for this project. When I asked 'How much is available?', I got "Well, I don't know, but, you know, there is always some end of the year money that we have to spend".
Sooooo, hit me with your best shot. I am thinking of a simple stereo scope, a good fiber optic light source, a simple digital camera, and a simple digital printer for 3" x 5" pictures. It will probably have to be on the inexpensive side and fairly bulletproof.
If you have any experience with specific combinations of components that worked well or turned out poorly, let me know and I will pass your info. on to the interested folks here.
Thanks again for your thoughtful contributions.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu **Area code changing to 831 as of 7/11/98**
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Recently there has been interest in providing TEM or SEM pictures of single celled creatures for use in educational websites, etc. But while today we have access to wonderful imagery undreamed of by our forbears, we should not overlook the wonderful achievements of the optical microscopists of the 19th century. So if you want:
DESMIDS, DIATOMS or RADIOLARIANS
I cannot suggest better than "ART FORMS IN NATURE" by ERNST HAECKEL, one of the greatest of the 19th century microscopists. This book of his drawings was published by Dover in 1974. All sorts of creatures, laid out in beautiful symmetric patterns (Maurits Escher, eat your heart out!) How high Haeckel's reputation was can be seen in this quote from "Life and its Beginnings" (1914) by Dr Helen Webb:
"There is really no end to the wonders which can be told of the varieties of little one-celled creatures. I can here give you only a few examples of them and their peculiarities. If we want to know more, as we all do, there are numbers of interesting books written about them which we can read. Even when the books themselves are very difficult the pictures are delightful to look at. Some of the loveliest are to be found in Haeckel's books, and if ever you have an opportunity I advise you to try and draw some of them.
O Lord, how manifold are Thy works ! In wisdom hast Thou made them all: ...."
There is really an irony here, because in Dr Webb's book we have a good Christian lady writing a gentle introduction to the facts of reproduction, while Dr Haeckel was a colourful and controversial character, notorious for his outspoken atheism - so much so that whenever he visited Darwin at Down House, everyone was on tenterhooks in case he came out with something over-the-top.
But Haeckel's contribution to microscopy was far greater than simply being a superb artist. Most notably, he brought to attention the features of embryonic development, for example the transitory gill-slits that appear in mammalian embryos. This he tended to over-interpret, as if the embryo were to go through a "fish stage", then a "reptile stage", before "evolving" into a mammal. The doctrine was stated as:
"Ontogeny recapitulates Phylogeny".
Haeckel has been recently criticized for seeing and drawing too much of "conserved stages" in embryonic development - in other words of seeing what he wanted to see under the microscope (long before the days of image manipulation!) On the other hand, it is argued that he got the broad picture right, and the details were sorted out later. I would say there is truth in both sides. Even with my own humble polymer spherulites, I realize that the developments I have seen under the microscope were not as regular as I at first saw or thought.
Anyway, do look at these drawings. So many "handbooks of pond life" published since must owe something to them. But I am sure that these micro forms have had an influence far beyond science, and that countless science fiction covers could trace back their ancestry to this source. But there is something even more - one is really seeing the micro world through the eyes of the 19th century. Looking at these drawings gave me a slightly vertiginous feeling, as if I had entered a fold in space-time and was really looking directly into the world of our Victorian ancestors.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Absolutely daft - but I have to ask you-all if anybody knows a source of copper gauze, so that we can make the kind of baskets on which we place grids for extracting replicas?
UK supplier would be most convenient, but I don't mind looking across the Channel or the Pond.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Micro-tools' phone number is (408) 395-1585. The part is Cat. #S-078-B and they call it a fine point diamond scribe. They should be considered expendable. This is the part number that Stacie Kirsch uses at Electron Microscopy Sciences also. I don't know what the South Bay Technology's number is. Get a minimum of 25 if you think you are going to use the technique regularly or maybe ten to try it out. When they break, you can use them for marking and for the coarse scribing step on the back side of the sample. You really need good sharp ones for the final cleaving step. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
----------
There is a company called Buckbee Mears that sells and makes all types of etched and electroformed meshes and masks in different sizes and materials. They will make items to order. Sorry, I don't have the number or the address. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Absolutely daft - but I have to ask you-all if anybody knows a source of copper gauze, so that we can make the kind of baskets on which we place grids for extracting replicas?
UK supplier would be most convenient, but I don't mind looking across the Channel or the Pond.
+----------------------------------------------------------------------- -+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley |
A short time back someone inquired about vibration isolation systems for EMs. With that in mind, I would suggest that you contact the following company:
Minus K Technology 420 S. Hindry Ave. Unit B Inglewood, CA 90301
They have some really great products. I have no financial interest in th= e product or the company - I've just seen the products and think they're pretty nifty.
To the CLSM users, Is there anyone out there using the Noran Oz Confocal system, and is doing at least double (hopefully triple label) fluorophore detection and are experiencing the following bugs: 1)If you shut down acquisition, the previous slit numbers are no longer optimal when you restart acquisition. 2)If you change the slit position numbers, the image shifts slightly (triple 0.5 um fluorescent bead overlays confirm this), which means recalibration almost everyday. 3)Uneven illumination across the field (seen especially with sections that encompass the whole field. 4)Incompatible astigmatisms especially the 633nm or cy-5 5)RF noise as seen as a horizontal line on the right hand side when the backround is high. These are our major concerns and eats up a lot of my time with constant recalibrations and checks. The company claims this is not unique to our system and are working on the problems. The also claim no one is using the system to the full capacity like we are (ie double and triple label). Has anyone seen the last demo at Woods Hole Mass? The system supposedly had a good showing. Did they use 0.5 um beads or huge pollen grains for demo. Did they do double and triple label imaging? Am I alone in my problems or are there others out there? Let me know.
Following up on an earlier posting: the Buckbee-Mears company referenced as a supplier of etched meshes etc is located in St. Paul, Minn and their phone number is 612 228-6434. -- Fred Schamber ....................... mailto:fhscham-at-SGI.NET
Hello All, I am interested in replacing an existing heating stage, (Leitz 350) which has served us well for } 10 years. If anyone has experience/comments on an appropriate replacement, ie. heating to 350 - 500C and will fit a Zeiss Axioplan microscope I'd like to hear from you.
Hi everybody, =20 we need a glow discharge unit to render carbon-coated grids=20 hydropohilic prior to negative staining. Does anyone have a) a machine= =20 which is not in use any longer that could be purchased at a reasonable= =20 price, or b) informations about companies still producing such=20 devices?=20 =20 Many thanks in advance for help, Matthias
Thank you. We at Electron Microscopy Sciences do sell Glow Discharge units stand alone as well as attachments to carbon coating units.If you send us your address we will be more then happy to give you all of the information on the units we have to offer. Thank you.
POSTDOCTORAL RESEARCHER IN ELECTRON MICROSCOPY OF DEFORMED ROCKS
Initial salary within the range =A315,462 - =A317,266 pa=20
A NERC-funded post is available for 2.5 to 3 years, from 1 October 1998 in = the=20 application of frontier electron microscopy techniques to examining deforme= d=20 rocks. In the Microstructure Research Group we have pioneered the use of=20 electron backscatter diffraction patterns, SEM orientation contrast imaging= and=20 TEM to characterise geological deformation microstructures and hence mechan= isms=20 and rheology. The post would suit geologists, materials scientists and=20 physicists with experience of electron microscopy and the ambition to devel= op=20 innovative techniques with applications within and outside Earth Sciences.
Informal enquiries to Dr J Wheeler email: johnwh-at-liv.ac.uk or Dr D Prior em= ail:=20 davep-at-liv.ac.uk. Web site=20 at http://www.pcweb.liv.ac.uk/johnwh/microstructure.html=20
Quote Ref: B/950 Closing Date: 17 July 1998=20
Further particulars and details of the application procedure may be request= ed=20 from the Director of Personnel, The University of Liverpool, Liverpool L69 = 3BX=20 on 0151 794 2210 (24 hr answerphone) or via email: jobs-at-liv.ac.uk=20
Web site at http://www.liv.ac.uk=20
John Wheeler Dept. Earth Sciences, Liverpool University, Liverpool L69 3BX, U.K. Email: johnwh-at-liverpool.ac.uk Fax: 0151 794 5170 Telephone direct: 0151 794 5172 Departmental home page on http://www.liv.ac.uk/earth_sciences/dept/general.html
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There have been requests for a summary of replies I received regarding image analysis packages. I usually delete e-mails shortly after receiving them, but here is the summary based on hard copies of e-mails I printed (recall that my application was grain sizing, so I may have disregarded packages that had primarily biological applications):
Image Pro Plus with Materials Pro plug-in: http://www.mediacy.com Princeton Gamma-Tech's Imagist: http:// www.pgt.com John Russ' Image Processing Toolkit: http://members.aol.com/ImagProcTK/index.htm Soft-Imaging Software Corp.: http://www.soft-imaging-web.de/products EMS On-Line: http://cimewww.epfl.ch/ Impuls's Vision XL/XXL: http://www.datex.de KS series from Zeiss: http://www.imas.co.uk/rep/kont/3czv.htm MetaMorph from Universal Imaging: http://www.image1.com Scott Scientific: http://www.scottscientific.com/ccad.htm Northern Eclipse from Empix Imaging: http://www.empix.com
Both Leica and Leco offer image analysis packages as well.
I also disregarded replies related to hardware, since we are only looking at software.
Hope this summary is useful.
Nancy Zjaba National Semiconductor South Portland, ME
by smith.edu (8.8.6/8.8.6/SC1.6) with ESMTP id IAA20050 for {Microscopy-at-Sparc5.Microscopy.Com} ; Thu, 18 Jun 1998 08:03:04 -0400 (EDT) Received: from SCIENCE/SpoolDir by science.smith.edu (Mercury 1.31); 18 Jun 98 08:03:05 EST Received: from SpoolDir by SCIENCE (Mercury 1.31); 18 Jun 98 08:03:03 EST
You can build on yourself quickly and inexpensively. See:
Aebi and Pollard, 1987, Journal of Electron Microscopy Technique 7:29-33.
We made one a number of years ago; I use it fairly often and it is terrific for just the task you mentioned. And when not in use in the glow discharge unit, the Tesla coil is fun to play with, but you need to be careful.
On Thu, 18 Jun 1998 matthias.morgelin-at-medkem.lu.se wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi everybody, } } we need a glow discharge unit to render carbon-coated grids } hydropohilic prior to negative staining. Does anyone have a) a machine } which is not in use any longer that could be purchased at a reasonable } price, or b) informations about companies still producing such } devices?
Matthias,
We bought a HDT200 `hydrophillic treatment device' from JEOL. It is simple to use and reliable. We use it to help prevent specimen contamination under the beam but it seems to be what you are after. I am unsure of the present cost but it was significantly less than other plasma devices available at the time.
I have no personal interest except as a satisfied customer.
Regards, Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
} Position: SEM Operator / Lab Technician } } Oz Technologies is a leading edge supplier of Test Interface products } for the Semiconductor industry, located in Hayward, CA. } } Description: } } Research and Development lab technician to perform variety of lab } related functions. Self-starter & Fast learner with an inquisitive and } exploratory mind to assist Engineers in Development of new products. } } Responsibilities: } } * Preparation and Cataloging of samples for SEM/EDS analysis, } * Executing Design-of-Experiments, } * Metrology assessment and Data Collection } * Administering Equipment (Operation, maintenance, calibration, etc.) } * Report generation and archival } } Skills required: } } * Proficient in running SEM (Variable Pressure) and EDS systems } * Proficient in cross-sectioning equipment, microhardness testers, etc. } * Ability to understand DOEs and instrument setup to execute } experiments } * Proficiency in computer skills (data collection, analysis, reporting) } is a must } * 4+ years of direct experience is desired } * Material Science (Metals, Polymers, Electro-Chemical) background is } a plus } * Measurement and Instrumentation background is a Plus } * Capability in handling multiple tasks with short turn around time. } } Organization } This position reports to Engineering Organization } } Additional notes: } Oztek is a dynamic, fast-paced company. As a result, employees are } periodically asked to perform duties outside the defined scope of their } position. We are a high tech company that manufactures } test and reliability products for the semiconductor, test and burn-in } industry. } Please visit our website for additional information about our company: } www.ozcorp.com } } If you are interesting in this position, please email me your resume. } Oztek reserves the right to change job functions as needed to meet } staffing requirements. }
} we need a glow discharge unit to render carbon-coated grids } hydropohilic prior to negative staining. Does anyone have a) a machine } which is not in use any longer that could be purchased at a reasonable } price, or b) informations about companies still producing such } devices? } } Matthias
You can do this cheaply and easily with a $50 Tesla coil. Arrange a good electrical contact with an unused feedthru in the baseplate of your evaporator, and discharge the coil during the rough pump part of the pumpdown. Or you can mount the coil on a vacuum dessicator and pump it with lab vacuum or a small rotary pump.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Several weeks back I requested information that would help educate the administration regarding the realities of "cost neutrality" at University microscopy facilities. I received about 35 responses 24 of which provided some information about how their labs were doing, the remaining 11 were interested in having a copy of the responses.
First, I am very grateful to those who took the time and responded. The compiled responses had a favorable impact on how some of the administration at my University perceived such a facility should be reasonably structured. I apologize for the delay but was waiting for some type of outcome regarding the future of my facility. I am in the process of making photocopies of the replies and mailing them to those who requested a copy. Those who requested anonymity will have any information regarding their identity removed.
There is clearly a trend at many Universities to attempt some cost recovery via a charge back system for microscopy facilities. This is not the issue, in my opinion. Microscopy facilities are expensive to run and some method for defraying the costs is a fiscally sound idea. However, based on the responses that I received, no US University generates revenue strictly from users to pay ALL of the bills (this includes all salaries for ALL staff, service contracts, repairs, supplies, etc). There were 2 non-US Universities who were able to do this but admitted that they have several large industrial contracts that made this possible. One of these still struggled every year to try to generate enough revenue.
} From those who provided such information, most microscopy (EM) facilities recovered from 20-40% of operating costs from users fees. Typically, 40-55K was generated in a year in larger institutions with several workers.
Clearly, most University microscopy facilities require subsidies in order to continue operation. A number of the facilities are fortunate to at least have salaries covered by their respective University and are only responsible for recovering service/maintenance and supplies (which is still no minor task). Others are not so fortunate and preside over facilities with ever shrinking support and an ongoing battle to justify continued subsidy. Some suggested that microscopy facilities should be considered the cost of doing business and that often such facilities are not given credit for the overhead generated from successful grant funding and money from students who take microscopy courses. Total cost recovery for microscopy facilities becomes dangerous when this is the primary focus and used as the sole basis for evaluation of the facilities success. Exploratory projects that have some risk of failure are too expensive to undertake since attempts to work out the most desirable protocol may (for example, immuno-gold labeling) still prove unsuccessful even when all technical precautions are heeded.
A few asked how I went from 0 K to about 65 K in one year. It wasnt easy. First I went to all the applicable colleges and gave a presentation on contemporary techniques in microscopy and molecular cytology. This seemed to generate a lot of interest. The rates were set at $50/hr beamtime and $25/hr for time I spent training students or preparing samples. This was affordable to most and I was quite busy. 5 months later I was required to raise rates to $75/hr beamtime (with me operating the microscope) and $50/hr preparation time. In order to offer grad students a less expensive alreally "multi-user" if only a few can afford to use it. I am addressing this issue with very good success by aggressively assisting researchers in writing the microscopy portion of their grants so that they can use the facility again. This has helped a lot. In addition, within the last several months we have incorporated a confocal microscope (which has been extremely popular) and an atomic force microscope into the facility. This is very beneficial because I can assist users with the most appropriate microscopy technique for their specific project.
In any event, the responses from a few dozen other facilities proved to have a very significant impact on at least some administrators and I am hoping that others can benefit as well. Please feel free to contact me if you have not already done so and would find the compilation useful. Also, please understand that my priority is still to keep things busy here so my response may be a bit slow but I will be happy to continue the dialog offline if anyone has more specific questions.
Best Regards,
Kirk J. Czymmek, Ph.D. Biological Electron Microscopy Facility University of Delaware Newark, DE 19716 kirk-at-udel.edu
On Wed, 22 Apr 1998, Kirk J C Czymmek wrote:
} } Dear Colleagues, } } I need information to educate the administration on how close to "cost } neutrality" a microscopy facility can get. I currently supervise a } facility with a part-time employee to assist in repairs. Much of the } equipment is 20-25 years old and is constantly breaking down. I am the } sole revenue generater with a few students who have been trained to use } the microscope and have beamtime hours. Within a year or so of operation } the facility went from generating $0.00 in revenue to ~$60K maybe } 70K/year. This doesn't pay all the bills of an EM Facility with 2 TEMs and } an SEM as well. My question is, is their any facility that is COMPLETELY } cost neutral with revenue only for service work provided? Are the numbers } I describe for a medium sized University reasonable? I would greatly } appreciate any input. If their is a University that spends only what they } bring in for doing samples in a multi-user environment please contact me I } be happy to know how you did this. } } Best Regards, } } Kirk J. Czymmek, Ph.D. } Biological Electron Microscopy Facility } University of Delaware } Newark, DE 19716 } kirk-at-udel.edu } } }
Regarding a good glow discharge unit, we also use one made very similarly to that mentioned by Dick Briggs and described in the paper: Aebi and Pollard, 1987, Journal of Electron Microscopy Technique 7:29-33. It works great and can be made in a minimally equiped shop for about $200.00. You will, however, need a vacuum pump (a considerable expense if you do not have one laying around already).
Good Luck!
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
Hello, I've been trying to embed brains from one day old rats perfused with 4% paraformaldehyde. The sections I've been getting have a number of cracks throughout the tissue. In particular, the cortex seems to separate from the rest of the brain. Below is a copy of the embedding protocol I am using. I would appreciate any suggestions/ modifications to the protocol which would improve the condition of the tissue. Thanks in advance-
Joanne
postfix for 16 hrs; PBS wash - 1 hr at 4 deg 0.85% Saline - 1 hr at 4 deg 1:1 saline/ 100% EtOH - 30 min at RT 70% EtOH (twice) - 30 min at RT 85% EtOH - 60 min at RT 95% EtOH - 60 min at RT 100% EtOH (twice) - 60 min at RT 100% Xylene (twice) - 60 min at RT 1:1 xylene/paraffin - 90 min at 60 deg. 100% paraffin (three times) - 20 min at 60 deg.
The objective stigmator for my Hitachi (H-300) transmission electron microscope has ohmed out and I am having problems locating another stigmator. Does anyone have an H-300 not in use that I might scavenge/purchase that part?
Thank you in advance,
Ginger
Ginger Baker EM Lab Manager OMS Secretary/Treasurer Research Dept., Oklahoma State University College of Osteopathic Medicine 1111 W. 17th St. Tulsa, OK 74107 Phone: (918) 561-8232 FAX: (918) 699-8629 http://osu.com.okstate.edu/dept/research/content/gbaker.htm lizard-at-osucom-fs02.ocom.okstate.edu
I apologize I just sent a posting regarding "Microscopy Facility Revenue Update" and while copying into my email a relevant portion was cut. For those who were interested and confused the following two lines are corrected below.
"$50/hr preparation time. In order to offer grad students a less expensive alreally "multi-user" if only a few can afford to use it. I am addressing"
SHOULD READ:
"$50/hr preparation time. In order to offer grad students a less expensive alternative they could be trained via an EM class or by me at $25/hr and then $50/hr if they operated the microscope without me. Once the rate change went into place most users could not afford to do EM. The facility was then being supported by 5 or 6 users with mega-grants and an occasional small EM project. This indicates that there is most definitely a threshold that one can charge users, and it becomes questionable whether a facility is really "multi-user" if only a few can afford to use it. I am addressing..."
Joanne: You don't indicate the size of your rat brain pieces which would make a difference in processing times. Basically I would think a longer processing time might be better 10 - 15 hours for the dehydration thru infiltration steps with an hour in each paraffin with vacuum applied in the last paraffin bath (58 degrees with polyfin) . If your tissue is brittle isopropyl alcohol would be preferable especially for the absolute alcohol steps. Butyl alcohol would harden (and shrink) even less when used in the absolute baths ( works wonders on muscle but takes longer to dehydrate and is very stinky). bob
We are in the process of purchasing an image analysis system for our department. We have narrowed our choices to the systems made by the following two companies:
-Buehler, Ltd. -Definitive Imaging
We need to decide on one of these two systems during the next week. Any comments about the pros & cons of each would be greatly appreciated. Also, I am not too familiar with the Definitive Imaging Company. Any information you can share about your experiences dealing with this company is also appreciated.
Thanks in advance, Viola L. Acoff, Ph.D. Assistant Professor The University of Alabama Department of Metallurgical & Materials Engineering Box 870202 Tuscaloosa, Alabama 35487-0202 phone (205) 348-3761 fax (205) 348-2164 Email VACOFF-at-COE.ENG.UA.EDU
I need some advice for the processing of very tiny wallaby embryos for TEM, without losing or damaging them. I have tried processing them from go to whoa in microfuge tubes with some success and also the V vials. I use an epon-araldite mix and it can be very hard to actually see the embryo when it is in the resin. I would really appreciate some help as these are the bane of my life TIA Joan Clark
In order to use a carbon coater w/o a thickness monitor and determine the thickness, it is important to have a highly polished brass or gold substrate as a calibration sample. I was interested in any information on manufacturers of these types of products or if any old polished brass item can be used. I have obtained info on the interference colors that correspond to different approximate thickness ranges, but in order to use this technique, there exists the need for a polished substrate as the calibration sample. Appreciate any information. Thanks, Steve
The Electron Microscopy Core Facility at the University of Missouri-Columbia has an open position for a full-time benefit-eligible senior electron microscopy specialist. Applicants should have a degree in Biology or a related field, and experience in the application of microscopy in the life sciences including plant, animal and microbial systems. Qualified applicants should have good communication skills to work with a variety of core facility users. These include faculty, staff, and students from a diversity of departments at the University.
Responsibilities will include sample processing, sectioning, and analysis with transmission and scanning electron microscopy, service of equipment, individual training and instruction for researchers, assistance in teaching a graduate laboratory course, and service to faculty and staff. Preference will be given to those demonstrating experience with immuno-electron microscopy. Applicants must also be able to supervise a full-time technician, and other part-time staff, coordinate general EM Core functions, such as scheduling user time, train others in the use of equipment, purchase and maintain equipment, maintain appropriate inventory of supplies, and participate in workshops to develop new skills. Basic personal computing skills are required. Salary is competitive and commensurate with experience. Position available immediately. Applications will be accepted until qualified candidate is identified. Please apply to:
Dr. Heide Schatten Associate Professor Department of Veterinary Pathobiology Director, Electron Microscopy Core Facility University of Missouri-Columbia Columbia, MO 65211 TEL: (573) 882-2396 FAX: (573) 884-5414 e-mail: vmhsch-at-showme.missouri.edu MU Electron Microscopy Core (EMC), Molecular Biology Program Web Site: http://www.missouri.edu/~emc/
Heide Schatten, Ph.D. Associate Professor Department of Veterinary Pathobiology Veterinary Medicine Building University of Missouri-Columbia 1600 E. Rollins Street Columbia, MO 65211 TEL: (573) 882-2396 FAX: (573) 884-5414 alternative e-mail: hschatte-at-facstaff.wisc.edu
Viola You seem to have already narrowed your choice to the two systems. However, in your initial survey, did you consider software by Scanalytics. They have a great image analysis program for both MAC and PC with additional support for gel analysis and 3-D. They were formally known as Signal Analytics but merged with another company last year to form the new Scanalytics. Their tech support is fantastic....I often get the founder and head of the company when I call for assistance....they all have amazing patience when helping guide the novice user through various procedures.
They can be contacted as follows:
Scanalytics 8550 Lee Highway, Suite 400 Fairfax, VA 22031 Phone: 703-208-2230 FAX: 703-208-1960 www.scanalytics.com
If you haven't made an absolutely final decision, do take a look at what they have to offer. They will have a booth at the upcoming MSA annual meeting in Atlanta.
Debby Sherman
++++++++++ Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
We are in the process of purchasing an image analysis system for our department. We have narrowed our choices to the systems made by the following two companies:
-Buehler, Ltd. -Definitive Imaging
We need to decide on one of these two systems during the next week. Any comments about the pros & cons of each would be greatly appreciated. Also, I am not too familiar with the Definitive Imaging Company. Any information you can share about your experiences dealing with this company is also appreciated.
Thanks in advance, Viola L. Acoff, Ph.D. Assistant Professor The University of Alabama Department of Metallurgical & Materials Engineering Box 870202 Tuscaloosa, Alabama 35487-0202 phone (205) 348-3761 fax (205) 348-2164 Email VACOFF-at-COE.ENG.UA.EDU
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Hi, We'd like to do some cryoSEM. Does anyone in the Atlanta area (or anywhere in the southeast US) have a cryostage on their SEM?
thanks, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Steven asks ... } } In order to use a carbon coater w/o a thickness monitor and } determine the } thickness, it is important to have a highly polished brass or } gold substrate } as a calibration sample. I was interested in any information on } manufacturers of these types of products or if any old } polished brass item can be used. ...
I sputter Au onto coverslips to the point of opaquecity (sp?), which I would guess is 80nM ... works perfect.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
First Circular for the 26th Scottish Microscopy Group Symposium
WEST PARK CONFERENCE CENTRE UNIVERSITY OF DUNDEE
Wednesday 11th November 1998
After the hugely successful 1997 =91SILVER ANNIVERSARY=92 of the=20 ScottishMicroscopy Symposium, we are now embarking for destination=20 =91GOLD=92. This years meeting will take place at the above venue and=20 the organising committee has arranged a Scientific Programme that=20 we hope will appeal to all microscopists.
The following topics will be presented by key-note speakers
Microscopy in studies of plant surface, structure and function=20 Chris Jeffree, Dept of Botany, University of Edinburgh.=20 =20 Apoptosis - when discretion is better than valour=20 David Harrison, Dept of Pathology, University of Edinburgh.=20 =20 Cathodal luminescence microscopy=20 Adrian Finch, Dept Environmental Sciences, University of Hertfordshire.=20 =20 Cryo Techniques=20 Alan Robbins, Oxford Instruments
These invited talks will be interspersed with short presentations and offer= s of these and/or posters can be made on the enclosed sheet.=20 A Second circular will be sent out in the near future containing the final programme and a registration form (registration cost =A320).
These meetings provide an opportunity to meet and share ideas with fellow= =20 microscopists. We look forward to seeing you.
For latest information visit our Web Site at http://www.abdn.ac.uk/~nhi691/= smg98.htm
Kevin Mackenzie EM Unit, Dept Zoology University Of Aberdeen Tillydrone Avenue Aberdeen AB24 2TZ Tel 01224-272847 Fax 01224-272396
} I was interested in any information on } manufacturers of these types of products or if any old polished brass item } can be used.
Anything you can polish should be OK. Try to choose something that you don't mind repeatedly cleaning. I ultimately had our shop mill me a brass cylinder that fit into our tilting-rotating stage so that it would be coated exactly as the stubs were, but anything that is at the same height and distance as the samples to be coated should do.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Dear Steve, Any old piece of brass will do, freshly polished with any metal polish or diamond 5 micron. I use a one inch diameter plug, 1/4 inch thick, others I know put their samples on a piece of brass sheet. I would hate to pay EM suppliers prices for such a simple item, but I'm sure Chuck Garber would sell you one.
You wrote: } In order to use a carbon coater w/o a thickness monitor and determine the } thickness, it is important to have a highly polished brass or gold substrate } as a calibration sample. I was interested in any information on } manufacturers of these types of products or if any old polished brass item } can be used. I have obtained info on the interference colors that } correspond to different approximate thickness ranges, but in order to use } this technique, there exists the need for a polished substrate as the } calibration sample. Appreciate any information. Thanks, Steve Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
What is the best scanner for digitizing high resolution transmission electron microscopy images? Both dynamic range and resolution are important to my application. Quality is more important than price to me. I would prefer HP scanners but they don't seem to make any high-end scanners.
Ching-Hwa Kiang
========================================== Dr. Ching-Hwa Kiang Visiting Assistant Professor Cram Teacher-Scholar Department of Chemistry and Biochemistry University of California Los Angeles, CA 90095-1569 Tel: (310) 206-0563 Fax: (310) 206-4038 Email: chk-at-chem.ucla.edu ==========================================
Rick Felten 06/19/98 05:05 PM If one examines the two exposed surfaces of a failed solder joint with SEM/EDS, is there a book or journal article that correlates the findings to a specific failure mode? Thanks Ric
pchouse wrote: } } I hope you get this message. It came from } http://msa.microscopy.com/MicroscopyListserver/FAQ.html } PAM } } --------------------------------------------------------------- } } Subject: vibration isolation tables } Date: Fri, 12 Jun 1998 20:01:34 -0400 } From: "Ron L'Herault" {lherault-at-bu.edu} } To: {microscopy-at-sparc5.microscopy.com} } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We have moved our lab (groan) and the new site is not ideal for our scanning } microscope. It looks like we need a vibration isolation table on the column } section. I'm looking for recommendations for good vendors. } } Thanks } } Ron } lherault-at-bu.edu I am unable to reach this person who sent a request appearently from your website of information . Messag received from receiving address is there is no such person. Your help would be greatly appreciated. Ted Cooper Scien-Tech Services / tcooper-at-sprintmail.com
A student at our university (Texas Christian University) working on her Ph.D. in neuroscience has been trying to stain dendrites using the Golgi-Cox technique. The primary reference she has been following is:
Kolb, Bryan and Jim McClimans, 1986, A process for cryosectioning of Golgi-Cox tissues, Stain Technology, 61, 379-380.
The main problem she has encountered is a difficulty in sectioning the tissue. Also, if and when a section is made it seems to dry out immediately upon contact with the slide.
Any suggestions or any other techniques you might know about would be most welcome. The student's name is Jocelyn Cooledge and her email address is: jjcooledge-at-delta.is.tcu.edu. Please reply directly to her.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I will be traveling outside the US (Europe, Australia, etc.) and want to = use my PowerBook 3400 Mac Laptop to communicate with back home and for = microscopy presentations. Question: Can one use ordinary power adapters from( 110 to 220 or = whatever) on the computer line? Do standard surge protects work or since = they are made for 110, would they be underrated? If the already = mentioned alternatives do not work, what does? Would appreciate hearing from the experienced microscopist travelers out = there in cyberspace. Thanks in advance, Judy Murphy
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
Can we have some feedback on what is involved in upgrading a JEOL 2000 series 200kV TEM from Freon to SF6?
An idea of cost would be of great interest.
TIA,
Mel Dickson ***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
I used to work for The Open University in England and I believe that there was some research work being carried out in to the above subject. For contact names and numbers see the Open University Web site: www.open.ac.uk.
The relevant information can be found in the Technology Faculty under the Materials Engineering Department.
In our vast knowledge base does anyone have any suggestions for making long edge glass knives, i.e. ~ 25mm or so as used for histology or rotary microtomes, with an LKB knife maker or glass breaking pliers?
Obviously, ideally we'd like to use a 'Ralph Knife' maker as sold by several vendors, but the $2k price tag is prohibitive presently (sorry guys) ... unless some has one gathering dust some where they'd be willing to part with....?
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
Dear Judy, I know that when a friend of mine tried that, the power conversion was the easy part, it was the phone connections that were much more difficult to resolve. Each country seems to have a different phone system and finding the converters isn't easy. Try Fry's or Radio Shack for a phone jack conversion kit.Good luck. The power was easy, since most computers have a switch to switch from 110v. to 220 v. You just need a plug converter, not a power converter. You wrote:. } } I will be traveling outside the US (Europe, Australia, etc.) and want to use my PowerBook 3400 Mac Laptop to communicate with back home and for microscopy presentations. } Question: Can one use ordinary power adapters from( 110 to 220 or whatever) on the computer line? Do standard surge protects work or since they are made for 110, would they be underrated? If the already mentioned alternatives do not work, what does? } Would appreciate hearing from the experienced microscopist travelers out there in cyberspace. } Thanks in advance, } Judy Murphy } } } Judy Murphy } Microscopy Technology Center } San Joaquin Delta College } 5151 Pacific Ave } Stockton, CA 95207 } 209/954-5284 } FAX 209/954-5600 } e-mail; jmurphy-at-sjdccd.cc.ca.us Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
There is a Hayat ref. to a paper on making Ralph knives by hand: Bennett et al., (1976) Stain Technology 51:71
I haven't tried it; don't even have the paper, but I'd like to hear from anyone who has as to how successful they were.
Tamara Howard CSHL
On Mon, 22 Jun 1998 edelmare-at-casmail.muohio.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } In our vast knowledge base does anyone have any suggestions for making long edge glass } knives, i.e. ~ 25mm or so as used for histology or rotary microtomes, with an LKB knife } maker or glass breaking pliers? } } Obviously, ideally we'd like to use a 'Ralph Knife' maker as sold by several vendors, } but the $2k price tag is prohibitive presently (sorry guys) ... unless some has one } gathering dust some where they'd be willing to part with....? } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "WE ARE MICROSOFT. } RESISTANCE IS FUTILE. } YOU WILL BE ASSIMILATED." }
I have not traveled there, but am familiar with theoretical requirements...
Depending on the circuitry and specs, some voltage converters should work. A "real" 220 to 110 (or 240 to 120) transformer should be used. Be sure to use any 120vac surge supressors on the 120 side NOT the 220 side of the transformer. ...The plugs won't match anyway...
Also, check the frequency, (unsure, but) you may be dealing with 50 Hz. rather than 60 Hz. Most modern computer switching power supplies will never know the difference, but the converting transformer should be rated accordingly. Operating a 60 Hz. (only) transformer at 50 Hz. can cause overheating.
A large transformer should not be needed. The VA rating should be about 1.4 times the computer power consumption in watts.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I will be traveling outside the US (Europe, Australia, etc.) and want to use
my PowerBook 3400 Mac Laptop to communicate with back home and for microscopy presentations. Question: Can one use ordinary power adapters from( 110 to 220 or whatever)
on the computer line? Do standard surge protects work or since they are made for 110, would they be underrated? If the already mentioned alternatives do not work, what does? Would appreciate hearing from the experienced microscopist travelers out there in cyberspace. Thanks in advance, Judy Murphy
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
by EMAIL1.BYU.EDU (PMDF V5.1-10 #23832) with SMTP id {01IYJB413TPW98G9IP-at-EMAIL1.BYU.EDU} for Microscopy-at-sparc5.microscopy.com; Mon, 22 Jun 1998 11:51:08 MDT
We are going to be offering a new service in our lab -- that of = quantitative image analysis. We are trying to decide what would be a = reasonable charge for performing this service. Would other labs = (preferably academic) offering this type of service please contact me = with their rate schedule.
Thanks,
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D Michael D. Standing BYU Microscopy Lab e-mail: MDStandi-at-bioag.byu.edu =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D
First on the connector issue..... in Europe, at least where I have traveled, I use a double female jack (american configuration), and take the plug out of the phone. MOST places I have traveled seem to have the american mail connector on the phone end of the cord. Also saves one from climbing over dressers, under tables to get to the wall jack. I will not say, don't buy the International connector kit, but do also make sure to buy the double female USA style.
Now for MY problem. I will be traveling in the USA this summer.... have paper will travel (presenting at Inter/Micro 50), and I am thinking of taking my lap top with me for email home and to keep up with the local press on the web. I know the power or the phone jacks will not be a problem, but how does one get on the web with no connections to servers in the states. Once there I can use my servers on this side of the Atlantic.
Any ideas on this appreciated...
Shalom from Jerusalem, Azriel
On Mon, 22 Jun 1998, Mary Mager wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Judy, } I know that when a friend of mine tried that, the power conversion was the } easy part, it was the phone connections that were much more difficult to } resolve. Each country seems to have a different phone system and finding the } converters isn't easy. Try Fry's or Radio Shack for a phone jack conversion } kit.Good luck. The power was easy, since most computers have a switch to } switch from 110v. to 220 v. You just need a plug converter, not a power } converter. } You wrote:. } } } } I will be traveling outside the US (Europe, Australia, etc.) and want to } use my PowerBook 3400 Mac Laptop to communicate with back home and for } microscopy presentations. } } Question: Can one use ordinary power adapters from( 110 to 220 or } whatever) on the computer line? Do standard surge protects work or since } they are made for 110, would they be underrated? If the already mentioned } alternatives donot work, what does? } } Would appreciate hearing from the experienced microscopist travelers out } there in cyberspace. } } Thanks in advance, } } Judy Murphy } } } } } } Judy Murphy } } Microscopy Technology Center } } San Joaquin Delta College } } 5151 Pacific Ave } } Stockton, CA 95207 } } 209/954-5284 } } FAX 209/954-5600 } } e-mail; jmurphy-at-sjdccd.cc.ca.us } Regards, } Mary } Mary Mager } Electron Microscopist } Metals and Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } fax: 604-822-3619 } e-mail: mager-at-interchg.ubc.ca }
Dear Azriel, I'm afraid you are out of my league. Access to your e-mail is a problem that you can best discuss with your Internet Service Provider or the computer genius at your university. You wrote: } This is almost a reversal of Judy's message. } } First on the connector issue..... in Europe, at least where I have } traveled, I use a double female jack (american configuration), and take } the plug out of the phone. MOST places I have traveled seem to have the } american mail connector on the phone end of the cord. Also saves one from } climbing over dressers, under tables to get to the wall jack. I will not } say, don't buy the International connector kit, but do also make sure to } buy the double female USA style. } } Now for MY problem. I will be traveling in the USA this summer.... have } paper will travel (presenting at Inter/Micro 50), and I am thinking of } taking my lap top with me for email home and to keep up with the local } press on the web. I know the power or the phone jacks will not be a } problem, but how does one get on the web with no connections to servers in } the states. Once there I can use my servers on this side of the Atlantic. } } Any ideas on this appreciated... } } Shalom from Jerusalem, } Azriel Best of Luck, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Eunsung Park wrote: ================================================= Does anyone know about the conductive epoxy that can stand high T (~500C)? Thank you. ================================================= I have never heard of a conductive epoxy that would withstand that high of a temperature. However, sometimes our SPI Silver Paste Plus™, which comes in a metal squeeze tube, is mistakenly described as being an epoxy. Its main use, at such high temperatures is to "glue" a silicon water to a heater head for those doing thin film coatings research.
Full information is on our website.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} ---------- } From: Tamara Howard[SMTP:howard-at-cshl.org] } Sent: June 22, 1998 11:39 AM } To: edelmare-at-casmail.muohio.edu } Cc: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Histology Knives / Ralph Knife Maker ? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Now for MY problem. I will be traveling in the USA this summer.... have } paper will travel (presenting at Inter/Micro 50), and I am thinking of } taking my lap top with me for email home and to keep up with the local } press on the web. I know the power or the phone jacks will not be a } problem, but how does one get on the web with no connections to servers in } the states. Once there I can use my servers on this side of the Atlantic.
The answer is to use a terminal at a library/university or subscribe to a US ISP for the duration of your stay. No long term obligation is necessary. (This is not a plug for ATT World-Net but I use them throughout the US when I travel although I use my university ISP at home.)
Over the past several years we have been asked many times, particularly by researchers new to corrosion casting, which color Mercox they should use, red, blue or clear. Since they are all the same except for the color, it appears the choice should be a based on personal preference and application. If anyone has any thoughts on which color should be used when, please respond and we will pass the information on to Fred Hossler so he can present it at his vascular corrosion casting sympossuim at MSA'98 in Atlanta on July 13th.
Thanks in advance,
John Arnott Chairman
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
We're not into Macs in this department, but visitors from US and Japan usually don't have any problems with our 230V 50 Hz supply as the Mac plugpacks (those that I've seen, at least) will work on anything from 100V to 250V (darned clever, these people). Check in your manual. Australia and UK are 240V 50Hz.
Ritchie
} I will be traveling outside the US (Europe, Australia, etc.) and want to use my PowerBook 3400 Mac Laptop to communicate with back home and for microscopy presentations. } Question: Can one use ordinary power adapters from( 110 to 220 or whatever) on the computer line? Do standard surge protects work or since they are made for 110, would they be underrated? If the } lready mentioned alternatives do not work, what does? } Would appreciate hearing from the experienced microscopist travelers out there in cyberspace. } Thanks in advance, } Judy Murphy }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Has anyone used Image Space Software for looking at dual channel "single section" images. Specifically using the dual channel overlays with two colours. I may be missing something but I cannot get the dual or stereo applications to work with single images. Probably this is not possible with this software it seems to be looking for depth information.
Also which antifade agent do people recommend for fluorescent antibody probes for use with confocal.
} There is a Hayat ref. to a paper on making Ralph knives by hand: Bennett } et al., (1976) Stain Technology 51:71 } } I haven't tried it; don't even have the paper, but I'd like to hear from } anyone who has as to how successful they were. } } Tamara Howard
It works rather well. You score on your "standard" glass knifemaker, and break with hand pressure. Worth trying.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
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Mel, We recently switched from Freon to SF6. You need to get a new gun assembly, to run at a higher pressure (3Kg instead of 2), and an exchange tank (rebuilt and tested in Boston -- since the changes inside are fairly extensive). Takes 3 to 5 days and costs about $28K here in the United States. -Mike O'Keefe ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hello world, } } } } Can we have some feedback on what is involved in upgrading a JEOL 2000 } } series 200kV TEM from Freon to SF6? } } } } An idea of cost would be of great interest. } } } } TIA, } } } } Mel Dickson } } ***************************************************** } } Mel Dickson, } } Director. } } Electron Microscope Unit, } } University of New South Wales. } } Sydney NSW 2052 Australia } } } } Phone (+612) 9385-6383 } } Fax (+612) 9385-6400 } } Website {http://srv.emunit.unsw.edu.au} } } ***************************************************** --------------4B8531A61E8BCF780EED5966 Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for O'Keefe, Michael A. Content-Disposition: attachment; filename="vcard.vcf"
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Dear Azriel, Check with your ISP. They may be a GRIC partner (see http://www.gric.com/). If so you can access your email account from anywhere in the world using a local phone call.
Yours sincerely,
Paul Thomson Techncial Director Thomson Scientific Instruments Web page: http://werple.net.au/~tsi/ Fax: +613 9663 3680 Tel: +613 9663 2738
First Announcement S4G International Conference on Stereology, Spatial Statistics and Stochastic Geometry Prague, Czech Republic, 21 June - 25 June 1999
under the auspices of Academy of Sciences of the Czech Republic Czech Technical University Charles University Czech Society for Cybernetics and Informatics
International Advisory Committee: J. Cwajna ( Poland ) D. Jeulin ( France ) L. Kubinova ( Czech Republic ) B. Pakkenberg ( Denmark ) D. Stoyan ( Germany )
S4G'99 Next in the series of conferences on Stereology and related sciences held in the Czech Republic and in the Slovak Republic (CMMSIA'82, CASIA'88, 6ECS'93).
Scope: This conference will be a meeting of scientists interested in spatial data evaluation, stochastic modelling and geome1rical sampling. Participation of people from applied fields is strongly desired, including quantitative microscopy and image processing.
Main topics of the conference are: Application of Stereology and image analysis in life sciences and material sciences. Application of spatial statistics in geostatistics, environmental and other sciences. Stochastic geometry and random sets. Integral geometry and theoretical Stereology. Quantitative image analysis and mathematical morphology.
Program: The scientific program will consist of keynote lectures, oral and poster presentations. No parallel sessions are envisaged The proceedings of contributed papers will be published
Venue: All scientific activities will be hosted by the Institute of Physiology, Academy of Sciences of the Czech Republic, located in the southern part of Prague with public transport connection to the center of the city. Accommodation in hotels and student hostels in Prague will be offered and cultural events for participants and accompanying persons will be organized.
Registration and important date:: If you are interested, please, return the completed reply card to the secretariat fill in the registration form on the conference WWW page. Your camera-ready abstract not exceeding 200 words on A4 sheet with wide margins should be sent to the same address not later than November 15, 1998. Alternatively you can send MS Word 6 file (with True Type fonts included) as an attachment to E-mail. Then you will receive the second announcement till the end of December. The papers for the proceedings should be submitted by February 15, 1999. Final program will be distributed during May.
Adjoining Course: Serology in Plant Anatomy 27 June - 31 June 1999
(contact: albrecht-at-prfdec.natur.cuni.cz or kubinova-at-biomed.cas.cz)
Local Organising committee: Viktor Benes, chairman Jana Albrechtova Vratislav Horalek Jiri Janacek Marie Jirkovska Lucie Kubinova Jan Rataj Ivan Saxl
Conference Secretariat: Jiri Janacek Institute of Physiology Videnska 1083 142 00 Praha Czech Republic Fax: (4202) 44472269 Phone: (4202) 4752768 E-mail: janacek-at-biomed.cas.cz WWW page: http://www.biomed.cas.cz/s4g.html
The most Important dates:
Abstract: November 15, 1998
Papers: February 15, 1999
Jiri Janacek Institute of Physiology Videnska 1083 142 20 Praha Czech Republic
Registration form:
Name:
Inst/Dept:
Address:
Phone:
Fax:
E-mail:
Title of my oral/poster presentation:
Signature:
======================================================================== Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
Next year, 1999, it will be 50 years since Philips sold its first transmission electron microscope, the EM100. In view of this special anniversary, we are trying to locate the oldest working model of this type. If you have information that can assist us, please let us know.
Your help is very much appreciated.
Bert Heijligers
FEI / Philips Electron Optics Bldg. AAE PO Box 218 5600 MD Eindhoven The Netherlands Tel: +31 40 2766225 Fax: +31 40 2766587 Email: bjh-at-eo.ie.philips.nl {mailto:bjh-at-eo.ie.philips.nl} Website: www.feic.com
Does anyone out there have experience of conditions for cleaning(mostly)cross sectional semi/super conductor specimens with a South Bay plasma cleaner?
We are setting up a new Electron Microscopy lab and have a plasma cleaner. Up to now success has been hit or miss, I would like to hear comments from other labs that have purchased such equipment. Published data on this technique seem to use a much lower bias voltage than we can achieve at 10W, although the manufacturer has explained this as a component error in the early versions.
Regards
Alan W Nicholls, PhD
NB NEW PHONE NUMBERS
Mailing Address:- University of Illinois at Chicago Research Resources Center (MC 937) 835 South Wolcott Avenue Chicago IL 60612-7341 Fax: 312 996 0539
Location Address:- Room 110, Research Resources Center - East 845 West Taylor Street Tel: 312 996 1227
I used & made this type of Ralf Knife around 20 years ago.
1. Start off with a 3 X 1 inch slide and diamond score using a small set square at the desired position.
2. Place a wood toothpick on the opposite side of your score line offset from the line by say 2mm, this distance is critical and may need to be made larger or smaller depending on the profile you achieve. Take care and wear leather gloves and protective goggles, carefully press down on the slide and you should have your Ralf Knife.
3. To use the knife on a microtome, I placed a standard steel knife in the holder with a piece of sheet metal under the steel knife to act as a ledge to support the Ralf Knife. Make your Ralf Knife a few mm. longer than the distance from the edge of the knife to the ledge at the base of the knife.
4. To fix the Ralf Knife onto the front face of the knife, paraffin wax was used by slightly heating both knife and Ralf Knife, and then waiting to both cooled down.
I hope this information is of assistance to you, please say if you need anymore information.
Best Regards.
Alan Bright
Bright Instrument Co. Ltd. St Margarets Way Huntingdon PE18 6EB England
---------- } From: edelmare-at-casmail.muohio.edu } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Histology Knives / Ralph Knife Maker ? } Date: Monday, June 22, 1998 04:29 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } In our vast knowledge base does anyone have any suggestions for making long edge glass } knives, i.e. ~ 25mm or so as used for histology or rotary microtomes, with an LKB knife } maker or glass breaking pliers? } } Obviously, ideally we'd like to use a 'Ralph Knife' maker as sold by several vendors, } but the $2k price tag is prohibitive presently (sorry guys) ... unless some has one } gathering dust some where they'd be willing to part with....? } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "WE ARE MICROSOFT. } RESISTANCE IS FUTILE. } YOU WILL BE ASSIMILATED."
"Analytical Electron Microscopy of Oxynitride Glass Ceramics"
A post-doctoral position is available in the Division for Microscopy= and Microanalysis at Chalmers University of Technology within the framework= of the Training and Mobility of Researchers (TMR) Programme. Qualified candidates should have a Ph.D. in materials science, physics or chemistry, and a documented background in transmission electron microscopy.= It will be considered an advantage if he or she has some experimental experience= of any of the following techniques: energy dispersive X-ray spectroscopy (EDX), electron energy loss spectroscopy (EELS), convergent beam= electron diffraction (CBED) or high resolution lattice imaging. The applicant= must be a national of a Member State of the European Community. The starting date of the appointment may be discussed, and the appointment may= last up to 16 months.
The position is funded by the TMR networks research project "Structure= and Properties of New M-Si-Al-O-N Oxynitride Glass Ceramics (M=3DY,Ln)"= which is a collaboration between seven partners in the United Kingdom, Ireland, =46rance, Sweden and Belgium. The collaboration within the established network will give the post-doctoral fellow good contacts with a number= of research groups in Europe and a good knowledge of a variety of preparative / analytical / property measurement techniques relevant to the evaluation of new ceramic materials as well as other materials outside the field= of ceramics.
The aim of the research project is to determine crystal structures= and properties of currently uncharacterised new glass-ceramic phases= in yttrium and rare earth sialon systems. These materials are potential refractory grain boundary phases in Si3N4-based ceramics, and have an interest= also as refractory surface coatings (glazes) and as interface materials in nitride-oxide and nitride-metal joints.
The analytical transmission electron microscopy work carried out= at Chalmers will focus on the chemistry and structure of rare earth= sialon glasses and their crystallisation products, and on microstructural development during nucleation and growth processes. Microanalytical= work on glass ceramics subjected to creep testing and oxidation / corrosion experiments may also be carried out.
The major part of the microscopy work will be carried out on a Philips CM200 Supertwin transmission electron microscope (TEM) with a field emission gun (FEG) and surrounding interactive instrumentation. = The FEG TEM is equipped with the Gatan imaging filter (GIF) which produces= energy filtered electron images and diffraction patterns as well as electron energy loss spectra, a Gatan off-axis CCD camera for high resolution imaging and a Link Isis energy dispersive X-ray (EDX) system for qualitative and quantitative elemental analysis including the light elements. There are also other electron microscopes in the laboratory, e.g. a Jeol 2000-FX TEM/STEM/SEM and a CamScan SEM.
Applications should be sent to: Associate Professor Lena Falk, Department of Experimental Physics,= Chalmers University of Technology, SE-412 96 G=F6teborg, Sweden e-mail: lklfalk-at-fy.chalmers.se; fax: +46 31 772 3224; phone: +46= 31 772 3321
We recently had such an upgrade done for a 2000ES by JEOL USA. The cost was between $25K and $35K, You would have to get a price from JEOL in Australia or Japan. We have not experienced any gun instabilities since. Both the gun and HV tank were replaced, and the upgrade took less than a week.
David Gelles Structural Materials Research Battelle Pacific Northwest National Laboratory Richland, WA USA Tel: 509 376-3141 Fax 509 376-0418 E-mail: ds_gelles-at-pnl.gov
-----Original Message----- From: Vetrano, John S Sent: Monday, June 22, 1998 9:05 AM To: Gelles, David S Subject: FW: SF6 upgrades for JEOL TEMs
Dave,
I figured you could answer this question if you get the chance. His address is: M.Dickson-at-unsw.edu.au
Thanks, JSV ---------- From: Melvyn Dickson Sent: Sunday, June 21, 1998 4:59 PM To: Microscopy-at-Sparc5.Microscopy.Com Subject: SF6 upgrades for JEOL TEMs
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
Can we have some feedback on what is involved in upgrading a JEOL 2000 series 200kV TEM from Freon to SF6?
An idea of cost would be of great interest.
TIA,
Mel Dickson ***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} 3. To use the knife on a microtome, I placed a standard steel knife in the } holder with a piece of sheet metal under the steel knife to act as a ledge } to support the Ralf Knife. Make your Ralf Knife a few mm. longer than the } distance from the edge of the knife to the ledge at the base of the knife. } } 4. To fix the Ralf Knife onto the front face of the knife, paraffin wax } was used by slightly heating both knife and Ralf Knife, and then waiting to } both cooled down. } } I hope this information is of assistance to you, please say if you need } anymore information. } } } Best Regards. } } Alan Bright } } Bright Instrument Co. Ltd. } St Margarets Way } Huntingdon } PE18 6EB } England } } e-mail: Bright-at-dial.pipex.com } Tel: +44 (0) 1480 454528 } Fax:+44 (0) 1480 456031 } } ---------- } } From: edelmare-at-casmail.muohio.edu } } To: microscopy-at-Sparc5.Microscopy.Com } } Subject: Histology Knives / Ralph Knife Maker ? } } Date: Monday, June 22, 1998 04:29 } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } In our vast knowledge base does anyone have any suggestions for making } long edge glass } } knives, i.e. ~ 25mm or so as used for histology or rotary microtomes, } with an LKB knife } } maker or glass breaking pliers? } } } } Obviously, ideally we'd like to use a 'Ralph Knife' maker as sold by } several vendors, } } but the $2k price tag is prohibitive presently (sorry guys) ... unless } some has one } } gathering dust some where they'd be willing to part with....? } } } } } } Richard E. Edelmann, Ph.D. } } Electron Microscopy Facility Supervisor } } 352 Pearson Hall } } Miami University, Oxford, OH 45056 } } Ph: 513.529.5712 Fax: 513.529.4243 } } E-mail: edelmare-at-muohio.edu } } } } "WE ARE MICROSOFT. } } RESISTANCE IS FUTILE. } } YOU WILL BE ASSIMILATED."
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
Authenticated sender is {3qt3gt-at-worldnet.att.net}
I have a client who is looking for a SEM, with EDS, that can accommodate a cable sample approximately 2 feet long without cutting it as it is involved in litigation.
Does anyone have such an instrument (and operator) for hire?
Dear Alan, } } I have a client who is looking for a SEM, with EDS, that can accommodate a } cable sample approximately 2 feet long without cutting it as it is involved } in litigation. } } Does anyone have such an instrument (and operator) for hire? } Does your client need images, or just local compositions? In the latter case, perhaps proton-induced x-ray emission (PIXE) or x-ray fluor- escence (XRF) would be suitable. I know of facilities which have done PIXE on a Gutenberg Bible, so they should be able to accomodate the size of the sample. Good luck. Yours, Bill Tivol
Marian College in Fond du Lac, Wisconsin is looking for a good used TEM for biological imaging to replace their early-60's RCA EMU3G. The closer the better, the more robust the better, and the cheaper the better. Anybody have an old scope to unload?
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have a client who is looking for a SEM, with EDS, that can accommodate a } cable sample approximately 2 feet long without cutting it as it is involved } in litigation. } } Does anyone have such an instrument (and operator) for hire? } } If so, I will pass your name and number along. } } Thanks. } } Alan Stone } ASTON
A JEOL IC848 can easily be modified to accomodate a sample that large depending upon the sample thickness. email me if you need more info.
A few thoughts that come to mind: 1) Check with Amray for info on large chambers. They have built systems that can accomodate aircraft turbine blades. 2) If the cable is part of a failure investigation, then it may be worth the expense of X-raying the entire cable section. Non-destructive analysis can be done in real time and can be video taped. 3) Since failures begin at microscopic site(s), the root cause(s) may not be revealed until the cable has been dismantled appropriately. 4) Who let the lawyers in to the lab?!
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
What is a good software to use for SEM digital image analysis?
Thanks Kalpana
************************************************************************************ Kalpana S Katti, Ph.D. Department of Polymers and Coatings North Dakota State University Fargo, ND 58105 ph:(701)231-8410 fax:(701)231-8439 email:kkatti-at-prairie.nodak.edu ************************************************************************************
Question: I have acquired an old microscope at a flea market and would like to find out what it is. It has one eyepiece and two objectives. There are quartz crystals in the optical paths. It has a rectangular box shape with a front sliding cover to access a ceramic slabe work base. Looks like it could be a binocular disecting microscope or a comparator microscope. Having acquired it in western Kentucky it may be related to analysis of coal.
A plate on the back has: Bausch & Lomb Optical Co. Rochester, NY No. 6222 Pat. Appl'd I estimate is is from about the 1920s. My wife estimates it is from late 1800s. My goal is to get the patient information so I can replace the missing objective lenses and use it. Bausch & Lomb have not been helpful. The old patient books are not much use without knowing the year.
Email: BKurnett-at-tech.puhsd.k12.ca.us Name: Bill Kurnett (High School Teacher)
School: Del Oro High School
State: California
Zip: 95650
Question: Is there anyone who can lend a small capacity mechanical vacuum pump to Del Oro High School in Loomis, CA. The pump is needed for a low-tech beam tube which can be used to demonstrate focusing by a magnetic lens. Student Joseph Navarro and Andy Hill are working on this project under the direction of teacher Bill Kurnett.
M.E. Taylor Engineering, Inc. would like to announce its new web site to = the=0Amicroscopy community.
We can be found at:
www.semsupplies.com
Please take a browse and see how we may be of service or help to you and = your=0Acompany.
M.E. Taylor Engineering, Inc. 21604 Gentry Lane Brookeville, MD 20833 Phone: 301-774-6246 =95 FAX: 301-774-6711 =95 e-mail: Metengr-at-aol.com = =95 See us=0Aon the web at www.semsupplies.com
Dear Alan, We have a large chamber (14"x14"x8") SEM with light element EDS detector, variable pressure attachment, and digital data storage. We provide analytical services for industry and and legal community. Igor Ivanov RJ Lee Group SEM/EDS,TEM,AES,XPS,SIMS,GCMS,FTIR,ICP,AFM,XRD,EPMA Analytical Services 530 McCormick Str. San Leandro, CA 94577 510-567-0480 510-567-0488 FAX
On 06/23/98 15:48:14 you wrote: } I have a client who is looking for a SEM, with EDS, that can accommodate a } cable sample approximately 2 feet long without cutting it as it is involved } in litigation. } } Does anyone have such an instrument (and operator) for hire? } } If so, I will pass your name and number along. } Thanks. } Alan Stone } ASTON
Dear Microscopy Listserver, Please change my address from : cshannon-at-fda.net to: cshannon-at-nctimes.net Thank you for your attention. Sincerely, C Shannon
Does anybody have a new or tried and true suggestion for a TEM magnification calibration standard for a mag of 30KX using a CCD camera? I would like to have better than +/- 3% accuracy. Have tried catalase--not happy with it. Latex spheres seem to shrink about 20% (according to a 1968 paper). Diffraction gratings not acceptable--only two to three lines in the image.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
I'm required to inject some sheep at 2 hourly intervals with Brdu. Most papers refer to a "magic" figure of 5mg/kg body weight, so per injection I'll need 250mg of Brdu. We seem to have a few problems dissolving this quantity of Brdu in a reasonable amount of saline (ie 20mls or less, in 20mls concentration would be 4mol/l). . Any thoughts on improving solubility would be appreciated.
Regards Peter Smith AgResearch Wallaceville Upper Hutt New Zealand
One quick idea would be us use something like 10nm colloidal gold particles and calibrate an image at high magnification using the particle lattice spacings, then make a (nominal) 30kX image and measure bulk features to get the actual mag at 30kX. You might want to track the magnification at one or more steps down the scale as you approach 30kX, to get better accuracy in the measurement.
There may also be some layered semiconductor samples that have highly accurate and known layer spacings that can be imaged nicely at 30kX. It seems to me that I have seen a paper or two on this type of sample, so a literature search might pop up with something (if the listserver doesn't...).
Larry
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
John Wheatley wrote: ============================================ Does anybody have a new or tried and true suggestion for a TEM magnification calibration standard for a mag of 30KX using a CCD camera? I would like to have better than +/- 3% accuracy. Have tried catalase--not happy with it. Latex spheres seem to shrink about 20% (according to a 1968 paper). Diffraction gratings not acceptable--only two to three lines in the image. ============================================== If you are looking for something beyond the possibilities mentioned, we have had some good reports from persons using the newly introducted MAG*I*CAL(TM) Calibration Sample. You can get a good sense of what it looks like at magnification at URL http://www.2spi.com/catalog/standards/magical.html
It is a bit more expensive that most other "calibration" samples but it is quite good and seems to be sufficiently robust that it does not fall apart after being used a few times. And it certainly is not going to be effected by the beam.
The product is available from SPI as well as several of the other main suppliers of consumables and accessories to microscopy laboratories.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We are looking for a low cost solution to replace an EDS analysis system on a JEOL 840 SEM. The new system should make use of our windowless Link 5474 detector, should be PC based (running Windows NT), and have the option for digital imaging.
Please reply directly to this address and not to the listserver. Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
John, I would argue that using a grating might still work. That is, provided you get some feature that is recognizable at a lower mag, say 10K. The larger number of lines at the low mag will give you a statistically valid number for that mag. Then the size difference between that low mag and the higher one gives you the factor to calculate the higher mag. We've done it on our scopes in just this way for quite a while and it's quite accurate. Of course, with the CCD camera you will still need to know exactly what the size of your CCD pixels is. We ran into the same problem with our CCD camera (Gatan 679) (Sherman et al, Micron 1996, 27:129-139).
Hope this helps,
Jaap
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Wed, 24 Jun 1998, John C. Wheatley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anybody have a new or tried and true suggestion for a TEM } magnification calibration standard for a mag of 30KX using a CCD camera? I } would like to have better than +/- 3% accuracy. Have tried catalase--not } happy with it. Latex spheres seem to shrink about 20% (according to a 1968 } paper). Diffraction gratings not acceptable--only two to three lines in } the image. } } John C. Wheatley } Lab Manager } Arizona State University } Center for Solid State Science } PSA-213 } BOX 871704 } Tempe, AZ 85287-1704 } } } Phone: (602) 965-3831 } FAX: (602) 965-9004 } John.Wheatley-at-ASU.Edu } } }
Does any one have an old LKB grid stainer that can be used for parts. We would like to get one for a spare. Please let me know, Tom Baginski, USUHS, Bethesda, MD
I have used the Mag-I-Cal sample that John McCaffrey and crew came up with and which South Bay Technology distributes. This sample covers the full range of Mags available in the TEM. It is done very quickly and simply and it can be used to measure the rotation calibration of the diffraction pattern and the camera constant as well. I have even used it in a High resolution SEM but its use there isn't quite as easy.
Incidentally, they have the Guiness World Record for the World's smallest ruler.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Does anybody have a new or tried and true suggestion for a TEM magnification calibration standard for a mag of 30KX using a CCD camera? I would like to have better than +/- 3% accuracy. Have tried catalase--not happy with it. Latex spheres seem to shrink about 20% (according to a 1968 paper). Diffraction gratings not acceptable--only two to three lines in the image.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
A colleague of mine is using araldite to mount samples and then performing experiments in cyclohexane. We would appreciate any information or experience of the solubility of araldite in this medium (or any others).
Thanks in advance
Dr. Giles Sanders Laboratory of Biophysics & Surface Analysis School of Pharmaceutical Sciences Nottingham University
We are looking at buying a Philips 400. Does anyone out there have any specimen holders to fit it that they would be willing to part with? Especially, does anyone have a straining stage they would part with (please, please, please!)? Double-tilt and/or low-background holders would also be of great interest. If so, please e-mail me here today or tomorrow, or, next week, e-mail me care of inal-at-nmt.edu since I will be away on travel for a while, but the people here will be able to contact me.
Thanks in advance for your time and attention.
Gill Bond Dept Materials & Met. Eng. New Mexico Tech
One last reminder - next Wednesday, July 1, is the date of the Biologic= al Sciences Meeting of the Midwest Microscopy and Microanalysis Society. = The meeting will be held at the Blood Research Institute in Milwaukee. Jim=
DiOrio, Biological Sciences Director of MMMS, has put together a great program, including a workshop on digital imaging. Tables will be avail= able for vendor literature, and there will be poster boards for any one wish= ing to display a poster.
If you intend to eat lunch with us, please remember to return your me= nu selections by tomorrow (Friday, June 26) on the forms mailed to you.
For more information, you may contact me via E-mail at jane.a.fagerland-at-abbott.com.
I'm looking forward to seeing you in Milwaukee next Wednesday!
MICROSCOPY SOCIETY OF AMERICA Sustaining Members Breakfast Monday, July 13, 1998 at 8:00 am to 9:30 am Georgia World Congress Center Room 255 W
Invited Speakers
Ralph Albrecht, MSA President Kathleen B. Alexander, 1998 Program Chair Janet Woodward, 1998 LAC Chair Charlie Meshul, 1999 LAC Chair Stuart McKernan, MSA Bulletin Brian Skepton, Springer-Verlag Philip Lesser, MSA Business Office Mary Beth Rebedeau, The Rebedeau Group
The speakers will present brief updates on important current and future Society Activities.
The favor of a reply is required.
Please R.S.V.P. to Paul Fischione, Chair - Sustaining Members Committee as soon as possible at e-mail: Paul.Fischione-at-internetmci.com or by fax: 724-325-5443
Does anyone know the correct pH for Richardsons stain used for = semithins. We are trying to match slides of skin done somewhere else in = which the ground substance (collagen, etc) of the dermis stained a purplish red.=20 TIA Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
Hi everybody, =20 we want to buy a scanner which can be used for both 35 mm and 70 mm EM= =20 film as well as 81x100 mm plates, probably with different adapters (?).= =20 Software must be compatible with Windows 95/NT 4.0 (e.g Adobe,=20 Freehand...). The higher resolution the better (at least 1200 dpi), th= e=20 cheaper the better, as usual. Any helpful comments or experience? -=20 would be highly appreciated. Thanks a lot, =20 Matthias
The Naval Research Laboratory is conducting a continuing search for a Director/Manager of the Marine Geosciences Division's Scanning Transmission Electron Microscopy (STEM) research facility. An announcement for this position is provided below. This opening is only open to US citizens.
GENERAL: The Marine Geosciences Division, Naval Research Laboratory (NRL), Stennis Space Center, MS, is seeking to fill the position of Director/Manager of the division's STEM research facility. The successful candidate will be an accomplished transmission electron microscopist with a Ph.D. degree in a closely related academic field, or possibly Ph.D. equivalent.
FACILITY MANAGEMENT AND OPERATION: The successful candidate will serve as the director and manager of the facility and will be fully responsible for: (1) competent, efficient, economical, and safe operation and maintenance of NRL's 300 kV STEM with Environmental Cell and supporting instrumentation for imaging and analyses including Scanning Mode Microscopy (STEM), Energy Dispersive X-ray Spectrometry (EDS), Electron Energy Loss Spectrometry (EELS), a slow scan, charge-coupled device (CCD) camera and other data acquisition systems, and a 100 kV Transmission Electron Microscope with Environmental Cell; (2) staffing the facility with high quality support personnel; (3) setting up operating procedures and protocols for all aspects of work in the facility; (4) supervision and training of technical personnel; (5) setting up the financial operations of the facility; (6) setting up a cataloging/archiving system for samples, images, etc.; and (7) ensuring that operations conform to all NRL/Navy health and safety regulations and standards.
RESEARCH: As the senior electron microscopy expert, the incumbent will conduct forefront basic and applied research directed towards developing state-of-the-art microscopy techniques and applying these to investigations of complex problems in marine geomaterials, geochemistry, microbiology, environmental processes, and related areas. A primary focus will be directed towards Naval problems particularly as involve understanding fundamental processes associated with the formation, deposition, and alteration of cohesive marine sediments and the impact on the geoacoustic and geotechnical properties of the sediments and behavior of these sediments in the marine environment. The inclusion of the environmental cell in the STEM provides NRL with one of the unique capabilities in the world. With the STEM and Environmental Cell the incumbent will be able to conduct forefront research on gaseous and aqueous chemical processes at the molecular scale.
FOR FURTHER INFORMATION: Contact Philip J. Valent, Code 7401, Naval Research Laboratory, Stennis Space Center, MS 39529-5004, telephone: 228-688-4650, facsimile: 228-688-4093, or e-mail: phil.valent-at-nrlssc.navy.mil.
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
The chemical resistance (as well as temp resistance & strength) of epoxies, for a given resin, depends greatly on the curing agent and cure conditions used with the resin. I suggest consulting Ciba literature (e.g., http://www.gluguru.com/Tech_Menu/Ciba_Araldite_Table/ciba_araldite_table .htm)or technical reps to see if the resin/curing agent your colleague is using meets their needs.
-----Original Message----- From: Giles Sanders [SMTP:Giles.Sanders-at-nottingham.ac.uk] Sent: Thursday, June 25, 1998 1:04 PM To: Microscopy-at-sparc5.microscopy.com Subject: Araldite contimination
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
A colleague of mine is using araldite to mount samples and then performing experiments in cyclohexane. We would appreciate any information or experience of the solubility of araldite in this medium (or any others).
Thanks in advance
Dr. Giles Sanders Laboratory of Biophysics & Surface Analysis School of Pharmaceutical Sciences Nottingham University
Does anyone know if this John McCaffery calibration standard is NIST tracable or if it has been NIST tested. Russ
-----Original Message-----
I have used the Mag-I-Cal sample that John McCaffrey and crew came up with and which South Bay Technology distributes. This sample covers the full range of Mags available in the TEM. It is done very quickly and simply and it can be used to measure the rotation calibration of the diffraction pattern and the camera constant as well. I have even used it in a High resolution SEM but its use there isn't quite as easy.
Incidentally, they have the Guiness World Record for the World's smallest ruler.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Does anybody have a new or tried and true suggestion for a TEM magnification calibration standard for a mag of 30KX using a CCD camera? I would like to have better than +/- 3% accuracy. Have tried catalase--not happy with it. Latex spheres seem to shrink about 20% (according to a 1968 paper). Diffraction gratings not acceptable--only two to three lines in the image.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
At 10:05 AM 6/26/98 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20 (?).=20 } Software must be compatible with Windows 95/NT 4.0 (e.g Adobe,=20 } Freehand...). The higher resolution the better (at least 1200 dpi),= the=20 } cheaper the better, as usual. Any helpful comments or experience? -=20 } would be highly appreciated. Thanks a lot, } =20 } Matthias ******************* If any single piece of hardware would fit your needs, I would think that it would be a top end, high $$$, high resolution flatbed scanner. However you MAY have difficulty finding a single piece of equipment that can do a really good job on such a wide range of film sizes. I am getting good results scanning 3=BC" x 4" EM negatives on an Agfa SnapScan 300/600 flatbed scanner. And I am sure that it would do a good job on your larger film sizes. However, although I have not tried it yet, I doubt that the results from scanning 35mm format on a large flatbed like that would be satisfactory. On the other hand the Polaroid SprintScan 35 does an excellent job on the smaller 35mm format. So we have both machines. Yeah, I know....$$$$$! Joiner Cartwright, Jr., Ph.D. Assistant Professor of Pathology Baylor College of Medicine Houston, Texas U.S.A.
The "blues" in Richardson"s stains are combined with sodium borate. This brings the pH into a high alkaline level (about pH 12). Most pH meters cannot measure accurately such high readings. Furthermore, it is not that important. There is no one "correct" pH for LM stain for epoxy embedded tissues. Coloration depends upon the fixation, dehydration, processing, and type of tissue components. Most importantly it depends upon the type of epoxide used for embedding. Components of Epon substitutes vary between suppliers. This can make a huge difference in coloration. Faced with trying to match a certain coloration of tissues which have been processed in another laboratory is extremely difficult. Try the following: 1. Suspend the stains (I assume they are from the same company as the original!) in several phosphate buffers of different pH ranges. (5,7,9,11?) 2. Before attempting to do anything at all, soak the sections in warm water for about 30 minutes. 3. After staining be sure to soak the sections for 30 seconds in 75% alcohol, then in 100% for 15 seconds or so. 4. Mounting media may change LM coloration! Use a neutral one such as CYTOSEAL (Fisher).
5. Depending with what and how the sections were embedded, you may have to remove unbound monomers. Contact me if you need the method for that by telephone (303-871-3026). (AM is best).
I tore my hair out (which I can hardly afford) for two years staining semi-thin skin biopsies. I wrote a couple of papers as a result. The bottom line is that LM staining of epoxide embedded sections is extremely complicated!!!!!!!! Try the first 4 items mentioned first. You might also consider giving the stained sections a quick rinse in basic fuchsin. This imparts a pinkish-purplish additive to the final slide.
This really is fun, it just does not seem like it some days! Hildy
To clarify my previous question on Araldite - my collegue is using Araldite(R) Rapide and the similar slower curing version, but what he is really looking for is an adhesive of similar consistency to these but which are inert and preferably insoluble in cyclohexane.
I am having a problem with a new IBM E56 Aptiva Computer. After only about 5 or 10 minutes after first turning it on, and trying to connect to the Internet it crashed, and subsequently doesn't recognize the hard drive. Even the Aptiva Restore CD software is unable to format the drive, and restore the software on the drive.
Someone suggested to me that perhaps the BIOS was damaged by Netscape, but I don't know how to get at this BIOS to fix the problem, and it's a pain to bring it back to the store, (with the long delay in getting back up and running)
Is there anyone who understands this BIOS and who can advise me on how to repair the damage? This is my first experience with PC's runnning windows 95, and my first experience with a crash that I could not recover from. It would be nice to be more self sufficient with regards to computer crash problems, especially if they can be quickly and easily solved right there on the spot, especially with lab computers.
Thankyou, Garry
PS: it also seemed odd to me that IBM's idea for "recovery" is to format the hard drive and reinstall all the software. I would have thought it infinitately preferable to just restore the system software causing the problem, and leave the rest of the files alone.
The timing of this question is pretty good! We have been trying for quite some time to find the right person in NIST to arrange for some form of certification for this sample, because of the frequent requests for ISO-9000-type traceability. Three days ago we finally received a usable letter - the text is included below. The biggest problem for both us and NIST has been that the MAG*I*CAL sample is made from a single crystal of silicon, with the calibration marks being directly traceable to the silicon (111) lattice spacing. This spacing is directly observable on the sample itself through lattice imaging; i.e.: the sample is internally calibrated to a "God-traceable" standard, and NIST generally doesn't bother confirming intrinsic properties of materials. Certification auditors requiring NIST-traceable samples are generally most concerned about adequate paperwork, This recent communication with NIST gives us that paperwork, I believe, as NIST acknowledges the lattice spacings of silicon as being well characterized and documented, making the MAG*I*CAL sample a NIST-recognized sample. We'll be discussing the proper NIST term to use with the MAG*I*CAL sample when Rob Gettings gets back from a business trip in a couple of weeks. In the meantime, a copy of the NIST letter will be available for users of the sample for their certification requirements. If anyone requires a copy of the NIST letter right now, they should contact South Bay Technology, the world-wide master distributor of the sample (fastest), or the vendor from which the sample was purchased (will take longer). The South Bay contact is:
David Henriks South Bay Technology, Inc. 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA TEL: 800-728-2233 (toll-free in USA) TEL: 714-492-2600 (outside USA) e-mail: henriks-at-southbaytech.com
I hope this adequately addresses your question.
Cheers John
John P. McCaffrey Institute for Microstructural Sciences National Research Council of Canada M-50 Montreal Rd. Ottawa, Ontario K1A 0R6 Canada
NIST U.S. Department of Commerce NATIONAL INSTITUTE FOR STANDARDS & TECHNOLOGY
Robert Gettings Standard reference Materials Program Bldg. 202, Rm. 212, Gaithersburg, MD 20899
June 23, 1998
Dear Mr. McCaffrey:
I am sending this communication in regards to your question concerning the calibration of Transmission Electron Microscopes (TEM) using silicon lattice spacings. NIST does not currently supply a standard for calibrating the magnification scale of TEM's, nor for the crystalline lattice spacings of silicon.
The crystalline lattice spacing is an intrinsic property of a material. For pure silicon, the spacing has been well characterized and documented by the scientific community. It is known to 6 decimal places, and can be obtained from the CRC handbook of Chemistry and Physics among other references.
Related work by NIST:
NIST is currently working on SRM 640c Silicon X-ray Diffraction Powder. This material, when finished will provide certified line positions traceable to the SI definition of length.
NIST is also working on a new standard, SRM1990, for the lattice spacing of single crystal ruby. The material will be in the form of a 0.15 mm sphere, and is expected to be complete by the end of this year.
Thank you for your interest in NIST standards.
Sincerely, (signature) Robert J. Gettings Project Manager ----------
I have used the Mag-I-Cal sample that John McCaffrey and crew came up with and which South Bay Technology distributes. This sample covers the full range of Mags available in the TEM. It is done very quickly and simply and it can be used to measure the rotation calibration of the diffraction pattern and the camera constant as well. I have even used it in a High resolution SEM but its use there isn't quite as easy.
Incidentally, they have the Guiness World Record for the World's smallest ruler.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Does anybody have a new or tried and true suggestion for a TEM magnification calibration standard for a mag of 30KX using a CCD camera? I would like to have better than +/- 3% accuracy. Have tried catalase--not happy with it. Latex spheres seem to shrink about 20% (according to a 1968 paper). Diffraction gratings not acceptable--only two to three lines in the image.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
As an E.M. specialist and ex engineer using computers to run a business y= ou cannot help getting involved with these beasts.
I too have just had a BIOS problem which was solved in a way that may be appropriate to you?
I upgraded my hard drive to find that the old BIOS could not run the upgrade! The way round the problem was to contact the manufacturer's web=
site where there were a number of BIOS upgrades available. I was running= , or trying to run on version 1.4, down loading version 1.5 now has my baby=
running and we are both happy with life.
Good luck, its worth a try!
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Larry Allard suggested: } One quick idea would be us use something like 10nm colloidal gold particles } and calibrate an image at high magnification using the particle lattice } spacings, then make a (nominal) 30kX image and measure bulk features to get } the actual mag at 30kX. You might want to track the magnification at one } or more steps down the scale as you approach 30kX, to get better accuracy } in the measurement.
I agree. We have had similar standards made for us, with a combination of colloidal gold particles of known sizes placed on a grid. Please contact me back-channel for more information.
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
In response to the following question: =============================================== Does anyone know if this John McCaffery calibration standard is NIST tracable or if it has been NIST tested. Russ ================================================ A Certificate of Calibration accompanies each unit of the product. The complete statement, as well as extensive additional information about the product, can be found at URL http://www.2spi.com/catalog/standards/mag-cert.html
The introductory paragraph that is on the Certificate is the following: ++++++++++++++ With regard to the tracability and certification of the MAG*I*CAL™ calibration sample, each sample is grown on {001} oriented single crystal silicon, and all spacings on the sample are directly referenced to the cross -sectional (111) lattice spacing of silicon on the sample itself. This spacing is visible by lattice imaging on the sample itself, allowing each sample to be self-calibrating. Each unit comes with a numbered certificate, the text of which is included below. This certificate has been used for ISO 9000 certification, with the argument that the sample is self- calibrating against the (111) lattice spacing of silicon (a fundamental constant), and to our knowledge, this is the highest quality TEM calibration sample available anywhere in the world at this time. ++++++++++++++++++++++++++++++ I hope that this information answers your question.
Disclaimer: SPI Supplies has been distributing this product (as well as others) and full details are available on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I've seen this sort of problem with several newer computers. The source of the problem is not clear. In one case the FAT (file allocation table) was corrupted necessitating a reformat. The best solution I've found is to install Norton Utilities and follow their advice for creating rescue disks, running Image on startup etc. There are many resources for information on the Web that are more appropriate (than this Microscopy forum) for discussing this sort of general computer issue.
Dear Chris: I assume you are looking for: Moxtek (soft x-ray technologies) 452 West 1260 North Orem, UT 84057 Dr. Clark Turner 801-225-0930 801-221-1121 FAX moxtek-at-moxtek.win.net
Yours Igor Ivanov Senior Scientist RJ Lee Group, Inc. Analytical Services 530 McCormick Str San Leandro, CA 94577 510-567-0480
On 06/26/98 16:04:38 you wrote: } Sorry finger slipped on the keys. } Message should be Moxtek not Mostek. } Anyone have a phone number please. } } Chris Gilpin } Biological Sciences Electron Microscope Unit } G452 Stopford Building } Oxford Road } Manchester } M13 9PT } phone +44 161 275 5170 } fax +44 161 275 5171 } http://www.biomed.man.ac.uk/biology/emunit/emhome.html } } }
Offhand, I would be suspicious of the cooling fan/system. I have seen a number of computers get flaky once the processor overheated. My first experience of that type was after installing a 486-50 overdrive processor in a PS/1. (Nobody told me it needed a cooling fan.) Things come to a halt very quickly once the processor overheats, and it doesn't take all that long to warm up. 5-10 minutes sounds like plenty long enough. I have had the same experience with trying to drive a processor a little to hard, like when I was trying to push my 120 MHz up to 133 MHz. It didn't seem like much of a push, but the manufacturers have already well pushed to the edge so that there is no room for a cooling fan to fail
If there is a cooling problem, I would not be surprised if BIOS setup also fails to function. Is there anything on it that works? Can you boot from a floppy?
If it does need a refresh of the BIOS, many companies provide files that you can download from the internet. You use them to prepare a bootable floppy which is then used to "flash" the BIOS on the subject computer. It seems daunting, but I succeeded on updating the BIOS on an HP Vectra on the first try. Fortunately, it wasn't as dead as yours, but that really shouldn't matter.
Warren
At 11:00 AM 6/26/98 -0500, Gary Burgess wrote: } } I am having a problem with a new IBM E56 Aptiva Computer. After only } about 5 or 10 minutes after first turning it on, and trying to connect } to the Internet it crashed, and subsequently doesn't recognize the hard } drive. Even the Aptiva Restore CD software is unable to format the } drive, and restore the software on the drive. } } Someone suggested to me that perhaps the BIOS was damaged by Netscape, } but I don't know how to get at this BIOS to fix the problem, and it's a } pain to bring it back to the store, (with the long delay in getting back } up and running) } } Is there anyone who understands this BIOS and who can advise me on how } to repair the damage? This is my first experience with PC's runnning } windows 95, and my first experience with a crash that I could not } recover from. It would be nice to be more self sufficient with regards } to computer crash problems, especially if they can be quickly and easily } solved right there on the spot, especially with lab computers. } } Thankyou, } Garry } } PS: it also seemed odd to me that IBM's idea for "recovery" is to format } the hard drive and reinstall all the software. I would have thought it } infinitately preferable to just restore the system software causing the } problem, and leave the rest of the files alone. }
Are you referring to the limitation of the old computers to access disks larger that 540M without an enhancement to the BIOS?
If so, most large disks (used to) come with a BIOS extender to allow access of the whole disk. Perhaps manufacturers figure the old BIOS systems are all gone and such utilities are no longer needed. But if a BIOS upgrade is available for your system to allow large disk access, then that would certainly be the better way to go.
Warren
At 12:24 PM 6/26/98 -0400, Steve Chapman wrote: } } Hi Gary, } } As an E.M. specialist and ex engineer using computers to run a business you } cannot help getting involved with these beasts. } } I too have just had a BIOS problem which was solved in a way that may be } appropriate to you? } } I upgraded my hard drive to find that the old BIOS could not run the } upgrade! The way round the problem was to contact the manufacturer's web } site where there were a number of BIOS upgrades available. I was running, } or trying to run on version 1.4, down loading version 1.5 now has my baby } running and we are both happy with life. } } Good luck, its worth a try! } } Steve Chapman
I would like to discuss anyone's experience using Raman micro-spectroscopy to get chemical information on particles. We are exploring the possibility of using the technique to provide information on nitrates, sulfates, phosphates, carbonaceous particles, hydrated particles etc., to complement present characterization of micron-sized particles by SEM and EDS.
I would also be interested in user's experiences with raman microscope systems that are commercially available. Thanks to all respondents.
Bob Willis ManTech Environmental Technology, Inc 2 Triangle Drive Research Triangle Park, NC Email: rwillis-at-epamail.epa.gov
The messages following my comments are some dated email messages regarding the Polaroid Sprintscan 45. It will do what you want it to do.
I bought one and am fairly pleased with it. I had some problems with the vendor getting it to me; it took about 4-5 months even though Polaroid had them in stock (this was told to me by a Polaroid rep). When the unit came in, many of the parts were missing: manuals, SCSI cable, negative carriers, power cord, and SCSI terminator. It took about a week to get the essential parts to get it working. It took much longer to get the TEM negative carrier and I never did get the SCSI cable that was supposed to be in, but I had another and so it was not an issue and I let the matter drop.
The TEM carrier that they eventually sent me is not the glass one with the anti-Newton glass that is described somewhere below. It is a pressed metal piece with two magnets that fits into the 4x5 holder. It is not as convenient as the 4x5 holder and the magnets crop off a little of the image. John Warren recommends that the magnets be cut long way to avoid this, but I haven't done this yet. However, I think that they are close to a good negative carrier with this metal piece if they were to replace the spring loaded plate on the 4x5 that has a rubber mat around the edges of the 4x5 hole on it with a TEM negative sized plate. This then would come down on the insert plate that they are now using with the magnets and eliminate the magnets. I think that this arrangement would be as easy to use as the original 4x5 holder. I'm planning on having our machine shop make one. This plate is held in place by four posts, springs, and snap rings on the posts.
It works very well and it is very fast. I still have a little trouble setting the correct scan conditions. I think that their scan software needs some work, though. I have had trouble getting the right scan conditions on some negatives. I like to use a histogram to set up the conditions, but it only gives a few sporadic lines and I haven't figured out why this happens. It took a little while to get it set up. I have a umax flatbed scanner on the system and the Polaroid software had trouble recognizing the device even though all of the SCSI devices, including the 45, were on line. This seems to be a minor irritant with Polaroid scanners in general because another system down the hall has a Sprintscan 35 and a Polaroid flatbed. I got that software to recognize the flatbed on that system by turning off the Sprintscan35. I basically did the same thing on our system and turned the flatbed off and it finally recognized the Sprintscan45. We solved the problem when we found that the TWAIN source selected was our umax32 source instead of the umax16 source while the TWAIN32 was using the Sprintscan45 source. Now we select the TWAIN32 device as either the umax32 or the Sprintscan45 and we can leave both devices on.
The Sprintscan 45 doesn't have a timed out feature and the light stays on all the time. That's a bit of a problem because I like to leave this system powered up all the time.
Generally, I'm very pleased with the system and use it often.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Does anyone know if a TEM negative carrier is available for the Polaroid Sprintscan 45 Negative Scanner yet? Anyone having any experience with the carrier (if it exists) and this scanner, could you send me a little message on what you think of the system?
Thanks.
-Scott Walck
Walck-at-PPG.com
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
I have had many conversations regarding digital imaging with a wide variety of end users. The points made here have been validated in those conversations. I thought that the point of view from law enforcement might be of interest.
When a photographer or law enforcement agent testifies as to what is seen in a given image, they are attesting to the fact that the image is an accurate _representation_ of what they saw and photographed at a given point in time at a given location. Based on the perceived reliability of the witness, a judge or jury determines whether that testimony is accurate.
There has been much discussion in Law Enforcement circles about digital watermarks and the like that would 'disappear' if an image were manipulated. One well known digital camera manufacturer who indicated that they had a proprietary file format that could not be manipulated was corrected by a hacker. Given the right equipment and talent, an analog film image could be scanned in, manipulated and output back to film and the best experts in imaging could not be able to tell and they will admit that. Someday, it may be possible, but not today. So, my point is that any image, digital or analog, is only as factual or accurate as the integrity of the person representing it as such.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation "See What Develops" 4525 Leonard Parkway Richmond, Virginia 23221-1809 804 254 1011 804 254 1013 Fax warrenj1-at-polaroid.com
The PDC 2000/40 has a new(45 days old) list price of $1699 and the PDC/2000/60 has a list price of $1999. There has not been a price reduction on the DMC as of yet.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation "See What Develops" 4525 Leonard Parkway Richmond, Virginia 23221-1809 804 254 1011 804 254 1013 Fax warrenj1-at-polaroid.com
Right, prices seem to be dropping. I recently visited one web site advertising the PDC-2000 for ~$2500 only to visit again a few weeks later to see it reduced to ~$1600. I haven't seen a comensurate price drop in the DMC 2000 but that is probably because I haven't recieved a recent quote. Kevin Brent Smith University of Louisville Biology Dept.
-----Original Message-----
For the record, the Polaroid PDC-2000 is available for use either "tethered" or "untethered", the latter with onboard storage for 40 or 60 images depending on the model. You will then need to connect to the computer to download your images, though. Price of the PDC-2000 is now under $2K for all models.
Hope this helps your information gathering process. For more, get back to me or check www.polaroid.com
John, As you know, I bought the Sprintscan 45. You know that I had first contact you about this and had several inputs from the microscopy listserver. I had a very long delay in getting the thing to me and when it finally arrived, there were parts missing. You had me contact Dennis Lizier and he sent out the standard parts for the scanner (sans cable, but I already had one). However, I told him that you had said that the TEM negative carrier with the antireflection glass plates would also be sent to me. I haven't received it. I have left several messages with him about this, but I have not gotten a response or a call back. The original order went out in Jan. We have paid for the unit with the good faith that the carrier would be delivered. Am I going to receive the negative carrier? Can you please help me? -Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
we want to buy a scanner which can be used for both 35 mm and 70 mm EM film as well as 81x100 mm plates, probably with different adapters (?). Software must be compatible with Windows 95/NT 4.0 (e.g Adobe, Freehand...). The higher resolution the better (at least 1200 dpi), the cheaper the better, as usual. Any helpful comments or experience? - would be highly appreciated. Thanks a lot,
I am a computer guy. Working on them, building them and repairing them is the work that I do every day so I can squander my pay on my real love -- microscopes.
If your computer comes back after turning it off an setting for a while, the bios [basic in/out system] is not the problem. You are getting information to the computer and out of the computer. It sounds as if you have a cooling fan problem. The processor gets overheated and then refuses to work. Letting it cool will allow it to function for a few more minutes and then it will refuse to work again. Depending on the CPU, you may be beyond the point of needing to replace it already. Look for extended troubles down the line. You can put a diagnostic on it and watch it slow down as it overheats.
Get the largest fan and heatsink set up you can for the CPU. Make sure it is a a ball bering fan. Sleev fans freeze up too easily. We sell very nice ones for $8.00. I am not soliciting your business, but pointing out that you should not pay an arm and a leg for a good one. $12.00 should get you the fan and heatsink you need. The main reason for this problem is dirt. Clean out your machine every so often -- three times a year. If it sets on the floor -- keep a closer eye on it. There is a lot of dust down there.
I am sure I live a long way from you, and it is difficult to diagnose problems without looking at them. Any number of problems may be occuring. If the machine flakes out on you at about the same interval without being on the internet, it may be the CPU cooling system. If the machine only fails while on the internet, it probably is something to do with the modem or the phone line or the intenet.
You are undoubtedly "a man of science and technology", apply the scientific method to it. If you are a biologist who believes in evolution rather than creation, make up you own story and set out to prove your theories correct because surely faith in the manufacturer of the piece has nothing to say about its oporation parametes.
Best of luck, Gary. Let us all know how it turned out!!
I am not lover of Microsoft though I use their products because they are the best system going. I have tried others. Please don't blame them for the fact that you do not back up. This is a very inexact science still. Back ups are the most important detail that most people ignore. MS has a good backup utility - especially on '95. Either use that or tape or disk media or best of all; with the prices of hard drive dropping drastically, put another large drive in you system and use it to back up your files.
Another piece of advice, stay away from the proprietary stuff the next time around. They can make for some wicked expensive repairs. Go to a computer shop and have them build you a clone. Every piece in it is replaceable with bigger, better and newer hardware. You will be much better pleased with the service and repair turn arounds too.
I am preparing to make microscope slides of palm flowers and fruits. Some of the fruits are quite woody as are many of the developmental stages of the flowers. I had planned to use the old parafin method of microtome preparation but I have been advised to consider resin. My funds and resources are limited, but the resin method seems to have more advantages.
Questions: 1. Can I use a manual rotary Spencer 820 American Optical Corp Microtome on resin samples?
2. What are the drawbacks to using the resin method?
3. What are the drawbacks to using the parafin method (toxicity/results/time)?
Thanks in advance for any and all responces. Please reply to my email: crow-at-aloha.net
I will be sectioning palm flowers and fruits, some of which are almost woody. I had planned to use the old parafin method. The microtome I have in my lab is a rotary Spencer 820 (manual) , American Optical Corporation. Resin preparation instead of parafin has been recommended to me.
1. Can I use the above rotary microtome to section resin prepared palm fruit material?
2. What are the benefits vs down sides of resin/parafin
3. What is the best type of prep procedure for using resin (books/references/labs?)
Thanks in advance for any ideas and experience. Please reply to my email:
crow-at-aloha.net Mattie
_______________________________________________________________ Melany H. Chapin
To Reply Please Remove Spam-Proof "XXX" from return address.
ADDRESS: crow-at-aloha.net
After dark, all cats are leopards. (Zuni proverb) We will be known by the tracks we leave. (Dakota) When the legends die, the dreams end; there is no more greatness (Shawnee) Every animal knows more than you do. (Nez Perce) ________________________________________________________________
We were not quite ready to announce this NIST information as we still hav= e some clarification left to do, but now that the word is out, everyone see= ms to want/need a copy of "the letter". With that in mind, and because of t= he number of requests I have received already, I will be posting a copy of t= he letter on our web site (http://www.southbaytech.com) within the next week= =2E =
When I have the exact page address, I will post it here.
It seems from my inspection of the BIOS that it was indeed set OK, and that Netscape didn't screw it up. However, it still would not boot up. The store was kind enough to exchange the computer for a new one though, which I appreciated, since it was only 5 minutes out of the box until I had this problem. My suspicion is that there was a bad -very bad sector on the hard drive that caused this nightmare.
Anyway, I would like to thank everyone who took the time to give me some suggestions, some of which were to get a new computer. I've learned something anyway.
Garry
} ---------- } From: Terry D. Krueger[SMTP:tdkrueger-at-imation.com] } Sent: 29 June, 1998 08:28 } To: Garry Burgess } Subject: Re: Computer BIOS Problem } } If I was sitting in front of your computer it would be a lot easier. } However, here goes. } When you turn the computer on do you see the RAM memory test and all that } stuff? You should see a message that says "Press {Delete} to enter setup." } That will get you into the Bios. Then all you can do is check to see if } the correct parameters are entered in the Bios to match the specifications } of you hard drive. The specifications of your hard drive should be printed } on a label on the drive itself, so you may have to take the box apart and } look at it. First you must make sure that a hard drive is entered at all. } You should be able to select that somewhere on the left side of the screen. } Then you must enter all the parameters on the right side of the screen such } as # of heads, # of sectors, Meagbytes or Gigabytes. My experience is with } slightly older equipment and I am assuming that there aren't any major } differences in this IBM aptiva Bios. If everything looks OK, you may just } have a loose ribbon cable going to the drive. It would be quite surprising } to have Netscape screwing up your Bios. The purpose of your Bios is just } to tell the computer what kind of hardware you have and how to use it. } There is nothing in Netscape that I have heard of that could reprogram it. } } }
Has anyone ever observed anything similar to this:????If so how did you fix it???
I have a filament image vibration in one direction at 30-60hz in a JEOL 4000. It is very subtle. I have observed this for 3 years but the amplitude has increased such that we would like to pin down the source and correct it.
Since initially observing the problem we have had a gun change, several filament changes, the high voltage tank worked on so I do not see these as probable causes. Also I do not think these would give a one direction effect. The magnetic fields of computers, monitors, and step motors do not seem to have any effect on the filament image vibration.
I am thinking that the deflector circuits would be most suspect. Usual tests on the power supplies look good. Any ideas would be appreciated. -- Dr. Roseann Csencsits Electron Microscopy Center Building 212/C217 Argonne National Laboratory 9700 South Cass Avenue Argonne, IL 60439-4838 Phone: (630) 252-4977 Fax: (630) 252-4798
Dear Roseann, } } Has anyone ever observed anything similar to this:????If so how did you } fix it??? } Our HVEM has had similar problems, which we found were caused by mechanical vibrations--the column resonates at 20 Hz. We determined that the vibrations arose from many small sources, and when we fixed every- thing we could think of (including when the bearings for the air condi- tioning fans for our 45 story building were replaced), the vibrations lessened and finally are no longer a problem.
} I have a filament image vibration in one direction at 30-60hz in a JEOL } 4000. It is very subtle. I have observed this for 3 years but the } amplitude has increased such that we would like to pin down the source } and correct it. } Does your column resonate in the 30-60 Hz region? 60 Hz would seem at first thought to be electrical, but maybe not. We had someone come in with a vibration generator, and when he swept through the fre- quency range while I was looking through the binoculars, I could see a change from quiescence to large motion as the frequency went through 20 Hz.
} Since initially observing the problem we have had a gun change, several } filament changes, the high voltage tank worked on so I do not see these } as probable causes. Also I do not think these would give a one } direction effect. The magnetic fields of computers, monitors, and step } motors do not seem to have any effect on the filament image vibration. } } I am thinking that the deflector circuits would be most suspect. Usual } tests on the power supplies look good. } Any ideas would be appreciated. } -- Sounds like electrical causes have been checked out. If your instrument is shock-mounted, it will be sensitive to acoustic vibrations, so you may have to look into air conditioning vents, etc. Good luck; tracking down this kind of subtle effect--which may arise from several sources--can be devilishly frustrating. When you fix one source, there may be little or no observable improvement; only when you fix lots of things will you notice a gradual improvement. Yours, Bill Tivol
Does anyone have information about this stain? What is its use in TEM? What is the procedure in it's use? One of the project leaders thought it would be good to use but didn't know why. Thank you for your help. PAM
I am in a mild state of confusion regarding an immuno-EM experiment where it seems that the secondary antibody (GAM 5 nm) does not recognize the primary (mouse IgG) once the primary is bound to its corresponding antigen in tissue. We expect that the antigen is present with periodicity on fibrils in the connective tissue matrix. After the tissue is emersed in antibody, we see periodic decoration of the fibrils, which indicates to us that the antibody is bound. However, secondary antibody does not bind to the tissue. We are convinced that the secondary conjugate is not defective, as we use it for other experiments. Has anyone else experienced a situation where a secondary does not recognize a primary once the primary is bound to its corresponding antigen?
Many thanks for your consideration,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
My name is Steve Short. I work for Technifab Products, Inc., a manufacturer of cryogenic equipment located in Brazil, Indiana. I am researching the use of detector dewars in electron microscopes and other microscopy related equipment.
If you have any basic information regarding the fundamental operation of electron microscopes and more specifically the role of liquid nitrogen detector dewars, please notify me as to how I may obtain that information.
Tel. 812-442-0520 e-mail to sms-at-technifab.com Fax 812-442-0891
A quick comment on your difficulty setting scan conditions for negatives: Scanner software often applies a gamma correction for negative scans, apparently to compensate for the low contrast of negative film used in conventional photography. With slow programs, you can sometimes see the scanned image change as built-in lookup table is applied after the scan is complete. Also, your scanner control software might offer a choice of film settings (Kodak, Agfa, ...). That's another tipoff that the scanner software is applying a lookup table after scanning. TEM negatives often have very high contrast, and aren't really the kind of negatives the scanner programmers had in mind. A common result of scanning TEM film as negatives is severe posterization (loss of gray levels).
To avoid this effect, try scanning the TEM film as positive transparencies (you can invert the contrast with other controls in the scanner software, or in Photoshop. Also, in the positive transparency mode (for Microtek scanner software at least), there is a further option to force a linear gamma correction. I sometimes find this mode useful for scanning extremely high contrast negatives.
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA 99352 email thomas-at-mme.wsu.edu
---------- From: Walck. Scott D. Sent: Saturday, June 27, 1998 12:41 AM To: matthias.morgelin-at-medkem.lu.se; Micro Subject: RE: negative scanner -Polaroid Sprintscan45 and collected previou s messages.
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The messages following my comments are some dated email messages regarding the Polaroid Sprintscan 45. It will do what you want it to do.
I bought one and am fairly pleased with it. I had some problems with the vendor getting it to me; it took about 4-5 months even though Polaroid had them in stock (this was told to me by a Polaroid rep). When the unit came in, many of the parts were missing: manuals, SCSI cable, negative carriers, power cord, and SCSI terminator. It took about a week to get the essential parts to get it working. It took much longer to get the TEM negative carrier and I never did get the SCSI cable that was supposed to be in, but I had another and so it was not an issue and I let the matter drop.
The TEM carrier that they eventually sent me is not the glass one with the anti-Newton glass that is described somewhere below. It is a pressed metal piece with two magnets that fits into the 4x5 holder. It is not as convenient as the 4x5 holder and the magnets crop off a little of the image. John Warren recommends that the magnets be cut long way to avoid this, but I haven't done this yet. However, I think that they are close to a good negative carrier with this metal piece if they were to replace the spring loaded plate on the 4x5 that has a rubber mat around the edges of the 4x5 hole on it with a TEM negative sized plate. This then would come down on the insert plate that they are now using with the magnets and eliminate the magnets. I think that this arrangement would be as easy to use as the original 4x5 holder. I'm planning on having our machine shop make one. This plate is held in place by four posts, springs, and snap rings on the posts.
It works very well and it is very fast. I still have a little trouble setting the correct scan conditions. I think that their scan software needs some work, though. I have had trouble getting the right scan conditions on some negatives. I like to use a histogram to set up the conditions, but it only gives a few sporadic lines and I haven't figured out why this happens. It took a little while to get it set up. I have a umax flatbed scanner on the system and the Polaroid software had trouble recognizing the device even though all of the SCSI devices, including the 45, were on line. This seems to be a minor irritant with Polaroid scanners in general because another system down the hall has a Sprintscan 35 and a Polaroid flatbed. I got that software to recognize the flatbed on that system by turning off the Sprintscan35. I basically did the same thing on our system and turned the flatbed off and it finally recognized the Sprintscan45. We solved the problem when we found that the TWAIN source selected was our umax32 source instead of the umax16 source while the TWAIN32 was using the Sprintscan45 source. Now we select the TWAIN32 device as either the umax32 or the Sprintscan45 and we can leave both devices on.
The Sprintscan 45 doesn't have a timed out feature and the light stays on all the time. That's a bit of a problem because I like to leave this system powered up all the time.
Generally, I'm very pleased with the system and use it often.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
From: Walck. Scott D. To: Micro Subject: Polaroid Sprintscan 45 Negative Scanner- Date: Wednesday, November 12, 1997 6:51PM
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Does anyone know if a TEM negative carrier is available for the Polaroid Sprintscan 45 Negative Scanner yet? Anyone having any experience with the carrier (if it exists) and this scanner, could you send me a little message on what you think of the system?
Thanks.
-Scott Walck
Walck-at-PPG.com
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
From: alline.myers To: gls4590 Subject: Polaroid Sprintscan 45 Negative Scanner-TEM film carrier Date: Thursday, November 13, 1997 11:09AM
Scott,
Hi. We have a Polaroid Sprintscan 45 that I use for scanning TEM negatives. It's a great scanner, and scans both high res. images and diffraction patterns well. We took the 2x2-inch negative adaptor, the film carrier, and a TEM negative to the shop at NIST and had them make an adaptor for us. It works fine. I don't think Polaroid has made an adaptor for TEM negtives yet. Let me know if you need more info.
As you can tell, I'm at NIST now and no longer working for Dr. Hren, although I still do TEM on samples for him. I had heard you had changed jobs a few months ago. How do you like Pittsburgh?
From: alline.myers To: Walck. Scott D. Subject: RE: Polaroid Sprintscan 45 Negative Scanner-TEM film carrier Date: Wednesday, November 19, 1997 12:23PM
} } Do you ever have problems with optical density in terms of bright and dark } areas of the negative? How long does it take to do a TEM negative at full } resolution?
The optical density is 3.4 and so far I haven't had trouble with it --- can see faint details in diffraction patterns. The problem comes when I print to the dye-sub printer; it can't reproduce everything I can see on the computer screen.
I can scan a full B/W negative at 2000dpi in 2.5 minutes; this is around a 40MB image. You need 2.5 times the image size free on your scratch disk (in Photoshop) to scan an image. If I scanned the same negative at 4000dpi (max. possible), it would be a 160 MB file!!! I haven't tried this, mainly do to space limitations on my computer. Usually I scan a 2-inch square region of interest at 2000dpi, store that, then reduce the image to 400 dpi for further work and to 150dpi to print.
From: bozzola To: Walck. Scott D. Subject: Re: Polaroid Sprintscan 45 Negative Scanner-TEM film carrier? Date: Thursday, November 13, 1997 7:53AM
} Does anyone know if a TEM negative carrier is available for the Polaroid } Sprintscan 45 Negative Scanner yet? Anyone having any experience with the } carrier (if it exists) and this scanner, could you send me a little message } on what you think of the system?
Hi Scott,
I have been evaluating one for almost a month now and I am moderately pleased with it. It consists of a composite plastic frame with a single pane of anti-Newton glass onto which you place the negs. They suggest using a sticky paper tape (as Post-It adhesive) to ensure non-movement of the neg during scanning. I was very dubious about this arrangement but it has worked out surprizingly well. I feel that a "kit" should be provided consisting of the main frame, the glass and several plastic or metal inserts to act as negative carriers. That way, one could select the best combination of components for glassless or conventional scanning.
I am very pleased with the SprintScan. The resolution is good, the plug-in software (Macintosh) works well in concert with PhotoShop and is simple to use. Although I bought the unit without ever trying it out (gulp!) I was assured that I could return it if not satisfied. Well, we're going to keep it. I can not compare it to other scanners such as the Agfa but I feel that it is a fine piece of equipment and should serve us well. Although we have a Gatan camera on our TEM, the quality of the images are inadequate for publication. So, we have opted to generate conventional negs and to by-pass the darkroom by scanning and then dye-sub printing the prints. We have been using the Fargo printer several years now but eagerly await delivery of the new Codonics 1660 printer which should be out towards the end of December. We ordered it nearly a year ago but it has been slow to materialize since it represents a new technology (direct thermal line printing) as well as a conventional dye sub unit. I hope the wait is worth it, because we have had to refer many people to Kinko's for the best quality prints.
Judy Murphy was thinking of buying one also. You might contact her at either murphy-at-inreach.com OR murphy-at-ms.sjdccd.cc.ca.us
Contact me if you have more specific questions.
John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
From: colin.veitch To: gls4590 Subject: RE: Polaroid Sprintscan 45 Negative Scanner-TEM film carrier? Date: Thursday, November 13, 1997 1:52PM
Hi Scott.
We purchased the 45 not long ago and have found it very good. The holders that come with the system are almost the right size (for our negs) and we haven't had to "modify" or build new ones yet. I imagine that building the correct sized insert for the holder would not be too difficult.
The plug-in interface to Adobe Photoshop (we are running it on a PC) is good, with several different ways of altering scan characteristics ie brightness contrast etc..
Scan times are good, less than a minute for a 35mm up to around 5 minutes for a full sized (6cmx9cm) neg at 4000 dpi.
We have had a couple of minor crashes of the system but these occurred while the system was logged on to the network and scanning at the same time. No net logon, no problem!!
We've been happy with the system so far, just be warned that if you are going to scan large negs at high resolution you'll need lots of drive space and RAM. We have 128Mb RAM and with a full colour large neg you can notice a slow down in the system!!
I've no association with Polaroid or the agents in Australia, just a happy customer!!
Good luck,
Colin V.
Colin J. Veitch Instrumentation Scientist CSIRO Division of Wool Technology PO Box 21, BELMONT, Vic. 3216. Australia.
E-mail: colin.veitch-at-dwt.csiro.au
Tel: +61 (0) 3 5246 4000 Fax: +61 (0) 3 5246 4811
} Does anyone know if a TEM negative carrier is available for the } Polaroid } Sprintscan 45 Negative Scanner yet? Anyone having any experience with } the } carrier (if it exists) and this scanner, could you send me a little } message } on what you think of the system? } } Thanks. }
From: Hendrik O. Colijn To: Walck. Scott D. Subject: Re: Polaroid Sprintscan 45 Negative Scanner-TEM film carrier? Date: Thursday, November 13, 1997 8:28AM
Hi Scott,
I'm considering getting the SprintScan 45 too. Let me know what you think.
Thanks, Henk Colijn
} } Does anyone know if a TEM negative carrier is available for the Polaroid } Sprintscan 45 Negative Scanner yet? Anyone having any experience with the } carrier (if it exists) and this scanner, could you send me a little message } on what you think of the system? } } Thanks. } } -Scott Walck } } Walck-at-PPG.com } } Scott D. Walck } PPG Industries, Inc. } Guys Run Rd. (packages) } P.O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } } "The opinions expressed are those of S.D. Walck and not of PPG Industries, } Inc. nor of any PPG-associated companies." } Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
From: Pauline C. Yu To: Walck. Scott D. Subject: Re: Polaroid Sprintscan 45 Negative Scanner-TEM film carrier? Date: Thursday, November 13, 1997 8:41AM
I was just asked that very question the other day by my supervisor. We just purchased a sprintscan 45 and have taken a long time to get it set up. It came with a bunch of adapters(including a 2"x2.5" holder) but none in that special TEM size. I've considered just improvising a frame to fit into the 4x5 holder to hold the TEM negs; I also think I might be able to find someone who could just machine one for me. Maybe if enough people request it, Polaroid will make one, or some enterprising EM supply company will create one... As for the Scanner, it requires one of the newer SCSI boards, and a really up to the minute computer if you don't want to wait too long. We're running it on a WinNT4 with dual processors, and it's hardly a "sprint-y" scan. But as far as image quality goes, I've been rather impressed. The color is rather off, so it does require color management(though if you're doing just b/w TEM, it's nothing to worry about). The driver is integrated into whatever photo-manipulation program you have(they expect you to have Photoshop, but it works with Corel PhotoPaint and PhotoHouse too).
From: bozzola To: Walck. Scott D. Subject: Re: Polaroid Sprintscan 45 Negative Scanner-TEM film carrier? Date: Friday, November 14, 1997 2:58AM
} Thank you very much for the info. This was just what I was looking for. } One question: when you ordered it, how did you get info about the TEM } negative carrier? I see the SprintScan 45 advertised in a couple of mail } order catalogs and I'm sure they won't know what I'm talking about if I call } them.
What happened was:
When we received the SS 45 I quickly ascertained that a TEM carrier was needed and so I called the technical support guys asking if one was available. They didn't know what I was talking about and referred me to several other admin types who finally told me that none was available. I then asked if they would provide me with some of the components from the 4x5 carrier so I could have one fabricated and they flatly said no. End of discussion. Being a bit disturbed by this attitude, I posted a message to the MSA server complaining about this. About three weeks later I got a call from some VP at Polaroid who had just come from a round of golf with Charles Garber of SPI. Charlie relayed my complaint to him with the admonition that SPI might even sell the unit if it had a TEM carrier. Anyway, the VP said that they would be "working on a solution". Right .... I thought. Well, they actually did develop one and John Warren of Polaroid contacted me to see if I would evaluate one of the first designs (the one I described to you in the earlier note). I was supposed to return it quite some time ago, but it has been in use and I hate to go back to using the 4x5 glassless carrier for the TEM negs. So, I am hoping they will leave it with me ....... .
Anyway, a unit is soon to be released and if you check with John Warren at WARRENJ1-at-CLIFFY.POLAROID.COM he may have more info on the availability. Like I said, it works fine as is but with a few simple modifications it will be great. I'll probably get in trouble for telling you about this "prototype" but you can mention that it makes a difference in your decision to buy and maybe they will forgive my indiscretion.
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I have had many conversations regarding digital imaging with a wide variety of end users. The points made here have been validated in those conversations. I thought that the point of view from law enforcement might be of interest.
When a photographer or law enforcement agent testifies as to what is seen in a given image, they are attesting to the fact that the image is an accurate _representation_ of what they saw and photographed at a given point in time at a given location. Based on the perceived reliability of the witness, a judge or jury determines whether that testimony is accurate.
There has been much discussion in Law Enforcement circles about digital watermarks and the like that would 'disappear' if an image were manipulated. One well known digital camera manufacturer who indicated that they had a proprietary file format that could not be manipulated was corrected by a hacker. Given the right equipment and talent, an analog film image could be scanned in, manipulated and output back to film and the best experts in imaging could not be able to tell and they will admit that. Someday, it may be possible, but not today. So, my point is that any image, digital or analog, is only as factual or accurate as the integrity of the person representing it as such.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation "See What Develops" 4525 Leonard Parkway Richmond, Virginia 23221-1809 804 254 1011 804 254 1013 Fax warrenj1-at-polaroid.com
From: WARRENJ1 To: Microscopy-at-sparc5.microscopy.com; Kevin Brent Smith Subject: Re[4]: digital camera (PDC-2000: Tethered vs. Untethere Date: Thursday, December 11, 1997 2:39PM
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The PDC 2000/40 has a new(45 days old) list price of $1699 and the PDC/2000/60 has a list price of $1999. There has not been a price reduction on the DMC as of yet.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation "See What Develops" 4525 Leonard Parkway Richmond, Virginia 23221-1809 804 254 1011 804 254 1013 Fax warrenj1-at-polaroid.com
______________________________ Reply Separator _________________________________ Subject: RE: Re[2]: digital camera (PDC-2000: Tethered vs. Untethere Author: Kevin Brent Smith {kbsmit01%homer.louisville.EDU-at-prdnet.polaroid.com} at INET Date: 12/11/97 12:40 PM
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Right, prices seem to be dropping. I recently visited one web site advertising the PDC-2000 for ~$2500 only to visit again a few weeks later to see it reduced to ~$1600. I haven't seen a comensurate price drop in the DMC 2000 but that is probably because I haven't recieved a recent quote. Kevin Brent Smith University of Louisville Biology Dept.
-----Original Message----- From: R-Brooks Corl [SMTP:CORLB-at-cliffy.polaroid.com] Sent: Thursday, December 11, 1997 11:44 AM To: Kevin Brent Smith; kszaruba-at-MMM.COM Cc: Microscopy-at-sparc5.microscopy.com; YONG SUN; CRAIG ARMSTRONG Subject: Re[2]: digital camera (PDC-2000: Tethered vs. Untethered)
For the record, the Polaroid PDC-2000 is available for use either "tethered" or "untethered", the latter with onboard storage for 40 or 60 images depending on the model. You will then need to connect to the computer to download your images, though. Price of the PDC-2000 is now under $2K for all models.
Hope this helps your information gathering process. For more, get back to me or check www.polaroid.com
From: WARRENJ1-at-POLAROID.COM To: Walck. Scott D. Subject: Re: Sprintscan 45 Date: Wednesday, May 13, 1998 10:04PM
Scott
As I had indicated, our product development was working on this and they have been promising it on a regularly basis. I forwarded your message and it seems that they are still working on a glass carrier. however, they have a solution in the meantime. Here is a clip of the message I got yesterday.
} } Yes, I have some good news. I have a new TEM film holder in my hands. This is a metal mask that fits into the 4x5 holder, just like the other masks. I have 9 to be exact and, for right now, they are for emergency sales only. We are making out a plan to make more and the number depends on the amount you think you need.
I am sending one to John Bozzola, our first SprintScan 45 TEM customer. He was very helpful with feedback. He said the glass holder was okay, but they preferred and open air solution because most of their picture taking process avoids the use of glass completely. He sounded quite excited about this holder.
Again, this mask works like the others. It is placed into the 4x5 holder and a magnet is required. This mask is easier to use because it has knobs on two sides of the holder enabling the user to place the film more square into the holder. My only recommendation is to cut one of the magnets in half; the long way, to use it properly.
This holder is not the glass holder we have been working on. The glass holder is in the last stages, and we should see something in the next couple of weeks. { { {
Dennis Lizier may have contacted you about this as well but if not he is getting one of these to send to you.
Thanks for your patience and support.
John
______________________________ Reply Separator _________________________________ Subject: Sprintscan 45 Author: "Walck. Scott D." {walck%ppg.COM-at-prdnet.polaroid.com} at INET Date: 5.11.98 7:15 PM
John, As you know, I bought the Sprintscan 45. You know that I had first contact you about this and had several inputs from the microscopy listserver. I had a very long delay in getting the thing to me and when it finally arrived, there were parts missing. You had me contact Dennis Lizier and he sent out the standard parts for the scanner (sans cable, but I already had one). However, I told him that you had said that the TEM negative carrier with the antireflection glass plates would also be sent to me. I haven't received it. I have left several messages with him about this, but I have not gotten a response or a call back. The original order went out in Jan. We have paid for the unit with the good faith that the carrier would be delivered. Am I going to receive the negative carrier? Can you please help me? -Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
---------- From: matthias.morgelin-at-medkem.lu.se To: Microscopy-at-sparc5.microscopy.com Subject: negative scanner Date: Friday, June 26, 1998 5:05AM
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we want to buy a scanner which can be used for both 35 mm and 70 mm EM film as well as 81x100 mm plates, probably with different adapters (?). Software must be compatible with Windows 95/NT 4.0 (e.g Adobe, Freehand...). The higher resolution the better (at least 1200 dpi), the cheaper the better, as usual. Any helpful comments or experience? - would be highly appreciated. Thanks a lot,
I'm required to inject some sheep at 2 hourly intervals with Brdu. Most papers refer to a "magic" figure of 5mg/kg body weight, so per injection I'll need 250mg of Brdu. We seem to have a few problems dissolving this quantity of Brdu in a reasonable amount of saline (ie 20mls or less, in 20mls concentration would be 4mol/l). . Any thoughts on improving solubility would be appreciated.
Regards Peter Smith AgResearch Wallaceville Upper Hutt New Zealand
The highest concentration I've been able to acheive in .9% saline is 12 mg/ml. That will ppt. when refrigerated, but goes back into solution thwn warmed and mixed. 10 mg/ml is the highest concentration that doesn't ppt. in the cold, and it mixes more quickly than 12 mg/ml.
Regards, Glen
On Tue, 30 Jun 1998, Smith, Peter wrote:
} I'm required to inject some sheep at 2 hourly intervals with Brdu. Most } papers refer to a "magic" figure of 5mg/kg body weight, so per injection } I'll need 250mg of Brdu. We seem to have a few problems dissolving this } quantity of Brdu in a reasonable amount of saline (ie 20mls or less, in } 20mls concentration would be 4mol/l). . Any thoughts on improving } solubility would be appreciated. } } Regards Peter Smith } AgResearch Wallaceville } Upper Hutt } New Zealand } } smithp-at-agresearch.cri.nz }
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; AFM Technology Leaders for Environmental / In Vitro AFM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
In my opinion you should think of: * the system contains free prim antibodies (prim Ab's which have detached from the antigen, or excess prim Ab's which have not been removed be a washing step). The free prim Ab's react with the GAM preventing the GAM to bind with the antigen-IgG-complex. * steric hindrance by a connective tissue component preventing the binding of the GAM to the prim. IgG. * presence of an other reagent for the prim. IgG's in the system binding first and occupying the binding sites for the GAM * incubation conditions which denature (conformational changes) the prim IgG
Check: * confirm the binding of the prim Ab's * incubate the prim Ab's and the GAM to interact with each other in an other incubation environment * perform the reaction using an other batch of GAM
Good Luck,
Marcel
------- Forwarded Message Follows -------
Dear Microscopists:
I am in a mild state of confusion regarding an immuno-EM experiment where it seems that the secondary antibody (GAM 5 nm) does not recognize the primary (mouse IgG) once the primary is bound to its corresponding antigen in tissue. We expect that the antigen is present with periodicity on fibrils in the connective tissue matrix. After the tissue is emersed in antibody, we see periodic decoration of the fibrils, which indicates to us that the antibody is bound. However, secondary antibody does not bind to the tissue. We are convinced that the secondary conjugate is not defective, as we use it for other experiments. Has anyone else experienced a situation where a secondary does not recognize a primary once the primary is bound to its corresponding antigen?
Many thanks for your consideration,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit Portland, Oregon 97201 DRK-at-shcc.org
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Has anyone ever observed anything similar to this:????If so how did you } fix it??? } } I have a filament image vibration in one direction at 30-60hz in a JEOL } 4000. It is very subtle. I have observed this for 3 years but the } amplitude has increased such that we would like to pin down the source } and correct it. } } Since initially observing the problem we have had a gun change, several } filament changes, the high voltage tank worked on so I do not see these } as probable causes. Also I do not think these would give a one } direction effect. The magnetic fields of computers, monitors, and step } motors do not seem to have any effect on the filament image vibration. } } I am thinking that the deflector circuits would be most suspect. Usual } tests on the power supplies look good. } Any ideas would be appreciated. } -- Hi Roseann,
Do you know where in the microscope the deflection is coming from? Is it in the same direction as one of the defectors (this can be checked)? Where is it in the column? The direction will change, due to the rotation of the lens, if it is above any lens you change. This can be used to locate the position of the instability in the column. I assume that the image is stable so it must be the gun or condensers. Any clue as to where to look will help.
One of our 4000s has an occasional problem that may be associated. Someone around here uses equipment that generates a field which seems to be picked up by the objective stigmator coils (or more likely the amplifier circuits). This affects one 4000 but not the other which is sited in the next room, I wonder if different amplifier circuits may be sensitive to particular frequencies. Have you got any field measuring system? If so can you match the frequency of the deflection to the frequency of an external field, even if it is within specification? Naturally by the time we get round to looking for the cause the equipment has been switched off, but if your problem is constant then you might have a chance.
Good luck Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
I want to take this time to thank everyone who responded to my recent question regarding preparation of wood composite samples for optical and electro-optic microscopy. It appears that the most viable suggestion is to use cryosectioning -- an option that I intend to vigorously pursue. Your help is appreciated.
Dear all, I am working on goat mammary gland involution. My problem is to find specific antibodies for Collagen IV, Collagenase IV, Upa and other costituents of basement membran, cross-reacting with goat tissues Could anyone help me? Thank you Silvia
Silvia Modina, PhD Institute of Anatomy of Domestic Animals Laboratory of Embryology and Animal Reproduction Faculty of Veterinary Medicine University of Milan Via Trentacoste, 2 20134 Milano, Italy Phone: (+39) 2 21.54.036 Fax: (+39) 2 21.40.745 E-mail: silvia.modina-at-unimi.it
At 12:24 am +0200 1/7/98, Henrik Kaker wrote: } As I know there is no such list server. } } Henrik } } geos-at-goldrush.com wrote: } } } } } } } Does anyone know of the list server for DM? } } } } George
As far as I remember, Digital Micrograph is a Gatan product. Why don't you look up Gatan at http://www.gatan.com/
Hope this helps
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Winnie Westbrook wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Please post this position description. } } #03171-Lab Specialist Senior - CISAT } Assists in all aspects of procedures required for the prepapration of } tissue for immunohistochemical analysis (whole mounts as well as } sections), fluorescent and light microscopy, transmission and/or } scanning electron microscopy. Competitive candidates must possess } knowledge of techniques for immunological staining, sectioning, mounting } tissue, evaluating sections and taking and developing publicaiton } quality photomicrographs. Applicant must possess demonstrated ability } to operate: ultramicrotome, vibratome, cryostat, and other TEM/SEM } equipment. Applicant will be responsible for maintenance of specimen } records, chemical supplies, reagents, etc in the histological laboratory } and for independently modifying lab procedures to obtain optimal } results. Interpretation of findings and ability to direct the work of } others also necessary. Four year degree in Biology or related field } supplemented by post-graduate experience in microbiology and/or } neuroscience preferred. Salary range: $24,337-37,995. } } Send resume and all correspondence to: } } Brenda M. Ryals, Ph.D } Professor } Communication Sciences and Disorders } James Madison University } Harrisonburg, VA 22807 } ryalsbm-at-jmu.edu
To All Who Applied:
All applications are being reviewed. The posting is now closed.
Are you certain that the primary can actually be recognized by the secondary, I mean do they match? You could use a dot spot test to see if they do (described in our Newsletter nr. 4). If the primary is recognized by the GAM-5 in such a test, you may have steric hindrance in your specimen. In that case using smaller particles may solve the problem. You could check for this by using GAM-FITC or the like.
Good luck, Jan
============================= Jan Leunissen, Ph.D. AURION ImmunoGold Reagents & Accessories Managing Director Costerweg 5, 6702 AA Wageningen The Netherlands
Smith, Peter wrote: } } I'm required to inject some sheep at 2 hourly intervals with Brdu. Most } papers refer to a "magic" figure of 5mg/kg body weight, so per injection I'll need 250mg of Brdu. We seem to have a few problems dissolving this quantity of Brdu in a reasonable amount of saline (ie 20mls or less, in 20mls concentration would be 4mol/l). . Any thoughts on improving solubility would be appreciated. } } Regards Peter Smith } AgResearch Wallaceville } Upper Hutt } New Zealand } } smithp-at-agresearch.cri.nz
Peter:
I dissolve 5 mg/ml Brdu without any problem in 0.9% saline containing 0.007N sodium hydroxide. I warm it in my hand for a few minutes. However, this concentration seems to be lower than what you are aiming for.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Dear Melany, your asking a lot here. There are so many considerations and variables to discuss in the paraffin vs. plastic question. Both techniques require a great deal of time for proper infiltration of plant tissues so that is not a factor. Ultimately, sections of tissue embedded in plastic (resin) have the potential to look better (ie. be thinner and allow better resolution if fixed properly). Paraffin is the way to go if you have a large project requiring survey type sections of hundreds of flowers or very large block faces as can be the case in a flower development study. 'Fairly' hard tissues can be sectioned in paraffin if the proper softening protocol is followed however palm fruits might be tricky in later stages.
I usually prefer to embed in plastic. There is no reason to use 'resin' (e.g. Spurr's (I don't recommend Epon for plant tissue BTW)) unless you are going to be doing TEM work. Some plastics like JB-4 (I think it's a methacrylate?) are easier to use, are less toxic and can be sectioned in much larger block faces at incredible thickness i.e. } 10 micrometers if you so desire. I have had good results lately sectioning JB-4 embedded flowers down to 1 micrometer thick with an Olympus rotary microtome. This requires however that you can equip your microtome with a glass knife holder. I have tried steel and tungsten knives with plastic and resin embedded tissue with no great success. If you already have a paraffin set-up you will find that most of it isn't used in the plastic embedding protocol and there will be a initial expense for plastic embedding accessories. Also, sectioning of 'hard' tissue may in fact be more difficult after embedding in plastic or at any rate wouldn't be too much easier. Woody tissues are usually a problem for the anatomist and the problem is probably best solved by treatment to soften the tissue prior to embedding in any media.
A few studies of parrafin embedded tissue have been published lately but their numbers are in decline. Some journals and reviewers seem to prefer the higher quality of sections/micrographs that are possible after plastic embedding. Ask yourself these questions as a guideline:
1) How many blocks to be sectioned? 10-100 plastic, } 100 paraffin.
2) Can a glass knife holder be obtained for the Spencer 820 or do you have access to an ultramicrotome? The glass knife limits the block face size to around 1/4 inch wide although see the recent discussion of Ralf Knife making techniques.
3) Can you afford the dollars required to convert to plastic? A example list of supplies that are different from the paraffin procedure would include: glutaraldehyde, buffer chemicals, vacuum pump and dessicator for infiltration, plastic embedding kit, polymerization trays and blocks for JB-4, glass knives and holder for microtome, and lots of sundry items.
Finally, the protocols for plastic and resin embedding are published in many places. Find any good book on plant microtechnique. The ones I always recommend are:
O'Brien, T.P. and McCully, M.E. (1981) The study of plant structure: principles and selected methods. Termarcarphi Pty. Ltd. Melbourne, Australia.ISBN 0 9594174 0 0.
Berlyn, G.P. and J.P. Miksche (1976) Botanical microtechnique and cytochemistry. Iowa State University Press, Ames, Iowa. ISBN 8138 0220 2
Good luck and write to me if you have other questions, John
} I am preparing to make microscope slides of palm flowers and fruits. } Some of the fruits are quite woody as are many of the developmental } stages of the flowers. I had planned to use the old parafin method of } microtome preparation but I have been advised to consider resin. My } funds and resources are limited, but the resin method seems to have more } advantages. } } Questions: } 1. Can I use a manual rotary Spencer 820 American Optical Corp Microtome } on resin samples? } } 2. What are the drawbacks to using the resin method? } } 3. What are the drawbacks to using the parafin method } (toxicity/results/time)? } } Thanks in advance for any and all responces. Please reply to my email: } crow-at-aloha.net } } Best wishes, } } mattie
________________________ C. John Runions, Ph.D. Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
geos-at-goldrush.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anyone know of the list server for DM? } } George
} As I know there is no such list server. } } Henrik } } geos-at-goldrush.com wrote: } } } } } } } Does anyone know of the list server for DM? } } } } George
A listserver for users of the DM Scripting Language does exist. Check http://www.public.asu.edu/~perkes/DMSUG.html for details.
Gert
==================================================================== Dr. Gert Oostergetel Phone: +31 50 363 4223 Biophysical Chemistry Fax: +31 50 363 4800 University of Groningen Nijenborgh 4 E-mail: oostergetel-at-chem.rug.nl NL-9747 AG Groningen The Netherlands ====================================================================
High, I'am studing osteoclasts cells culture "in vitro" on dentine slides in an Environmental Scanning Electron (Electrosan 2020). Is there anybody who has been working on this subject ? I need some recomendations (kV, Working distance, Pv H2O), or litaracy on this subject, because I didn't find anything. Thank you very much.
Dear all, does anyone know of a graphics package that can take several separate digital image files and montage them on a single page ready for printing?
Thanks,
Mark Munro
From Microscopy-request-at-MSA.Microscopy.Com Tue Jun 30 16:16:15 1998 Return-Path: Microscopy-request-at-MSA.Microscopy.Com Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id NAA22529 for dist-Microscopy; Tue, 30 Jun 1998 13:03:31 -0500 Received: from Pegasus.cc.ucf.edu (Pegasus.cc.ucf.edu [132.170.240.30]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id NAA22523 for {microscopy-at-Sparc5.Microscopy.Com} ; Tue, 30 Jun 1998 13:03:03 -0500 Received: from port14.3a.tserver.ucf.edu ([132.170.24.144]:2048 "EHLO [132.170.24.144]" ident: "NO-IDENT-SERVICE") by pegasus.cc.ucf.edu with ESMTP id {20503-14402} ; Tue, 30 Jun 1998 13:59:54 -0400 Message-Id: {l03130303b1bed89d6d25-at-[132.170.24.144]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
There is no need to switch from Freon to SF6. We were able to get 84 lbs of Freon for ~$1000. Freon is readily available if you can show that you are not using it as a refrigerant.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-pegasus.cc.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
There is no need to switch from Freon to SF6. We were able to get 84 lbs of Freon for ~$1000. Freon is readily available if you can show that you are not using it as a refrigerant.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-pegasus.cc.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Hello, everyone. Last week I asked for information about breaking glass Histo knives, and we are following up on the "freehand method(s)" recommended, but we have also been VERY fortunate enough to receive an LKB 2078 Histo Knife Maker as well!
Small problem: we do not have any instructions for the LKB 2078, and after having trashed a bunch of glass trying to 'figure it out' (obviously unsuccessfully), does anyone have a set of insturctions they could share with us? A fax would be great, or if you remember your own instructions a step wise summary (its doesn't look that complicated) would also be great.
Thanks!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
Dear Roeseann, I have seen something similar in my 200 kV Hitachi. It went away when I did a thorough cleaning of the upper accelerator rings. A tiny bit of charging something was causing the rapid deflection of the filament. I never saw the speck that did it, but after the cleaning hte filament was steady. You wrote:
} } Has anyone ever observed anything similar to this:????If so how did you } fix it??? } } I have a filament image vibration in one direction at 30-60hz in a JEOL } 4000. It is very subtle. I have observed this for 3 years but the } amplitude has increased such that we would like to pin down the source } and correct it. } } Since initially observing the problem we have had a gun change, several } filament changes, the high voltage tank worked on so I do not see these } as probable causes. Also I do not think these would give a one } direction effect. The magnetic fields of computers, monitors, and step } motors do not seem to have any effect on the filament image vibration. } } I am thinking that the deflector circuits would be most suspect. Usual } tests on the power supplies look good. } Any ideas would be appreciated. } -- } Dr. Roseann Csencsits
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
In response to the positive feed back from the dealers of Radical quality products in more then twenty countries. We have decided to open an office in Canada in order to offer wide range of Microscopes, Projectors, OHPs, Educational Laboratory Equipment and Accessories, to North America.
Recognize with various awards for growth and quality in International market at a very competitive prices, we also design and supply custom-built optical system or accessory for any specific need.
I look forward to hearing from you, for any further details on catalogue/pricelist or demonstration of our products.
Hi, I use IP Lab Spectrum to montage micrographs togather. It's a commercial software from Scanalytics. I have used Photoshop but it's slower.
Bob Underwood Derm Imageing Center U of Wash.
On Tue, 30 Jun 1998 mark_munro-at-bio-rad.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } does anyone know of a graphics package that can take several separate } digital image files and montage them on a single page ready for printing? } } Thanks, } } } Mark Munro } }
My research subject are sterile anthers of soyabean. I am mostly doing transmission electron micrographs, and am mostly interested in the tapetum cells during meiosis stages. While I was looking around in the tapetum I came across a nucleolus from which small discreet amounts of some material seemed to emerge (emerge rather then enter, because of the shape of the material). The material seemingly coming out of the nucleolus resembled cloud puff balls or smoke signals from a chimney. Has anybody ever seen such thing and/or knows even what it is???? mRNA, protein, else?
Tracey M. Pepper 1 Bessey Hall Bessey Microscopy Facility Iowa State University Ames, IA 50011 Phone: 515.294.3872 FAX: 515.294.1337 email: tpepper-at-iastate.edu
I posted this question for a graduate student currently working in my facility. Any input on this one??
Anybody working on nucleoli?
My research subject are sterile anthers of soyabean. I am mostly doing transmission electron micrographs, and am mostly interested in the tapetum cells during meiosis stages. While I was looking around in the tapetum I came across a nucleolus from which small discreet amounts of some material seemed to emerge (emerge rather then enter, because of the shape of the material). The material seemingly coming out of the nucleolus resembled cloud puff balls or smoke signals from a chimney. Has anybody ever seen such thing and/or knows even what it is???? mRNA, protein, else?
Tracey M. Pepper 1 Bessey Hall Bessey Microscopy Facility Iowa State University Ames, IA 50011 Phone: 515.294.3872 FAX: 515.294.1337 email: tpepper-at-iastate.edu
Silvia Modina wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear all, } I am working on goat mammary gland involution. } My problem is to find specific antibodies for Collagen IV, Collagenase IV, } Upa and other costituents of basement membran, cross-reacting with goat } tissues } Could anyone help me? } Thank you } Silvia } } Silvia Modina, PhD } Institute of Anatomy of Domestic Animals } Laboratory of Embryology and Animal Reproduction } Faculty of Veterinary Medicine } University of Milan } Via Trentacoste, 2 } 20134 Milano, Italy } Phone: (+39) 2 21.54.036 } Fax: (+39) 2 21.40.745 } E-mail: silvia.modina-at-unimi.it
Try the Devlopmental Studies Hybridoma Bank at http://www.uiowa.edu/~dshbwww or speak with Karen Jensen (319) 335 3826. She is extremely knowledgeable and helpful.
Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; AFM Technology Leaders for Environmental / In Vitro AFM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Compix software can montage the images to a vertical or horizontal strip or a best fit rectangle.
Patty Jansma Tel:520-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
On Tue, 30 Jun 1998 mark_munro-at-bio-rad.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } does anyone know of a graphics package that can take several separate } digital image files and montage them on a single page ready for printing? } } Thanks, } } } Mark Munro }
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; AFM Technology Leaders for Environmental / In Vitro AFM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; AFM Technology Leaders for Environmental / In Vitro AFM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
I am in a mild state of confusion regarding an immuno-EM experiment where it seems that the secondary antibody (GAM 5 nm) does not recognize the primary (mouse IgG) once the primary is bound to its corresponding antigen in tissue. We expect that the antigen is present with periodicity on fibrils in the connective tissue matrix. After the tissue is emersed in antibody, we see periodic decoration of the fibrils, which indicates to us that the antibody is bound. However, secondary antibody does not bind to the tissue. We are convinced that the secondary conjugate is not defective, as we use it for other experiments. Has anyone else experienced a situation where a secondary does not recognize a primary once the primary is bound to its corresponding antigen?
The problem is not that the secondary does not recognize your primary. It is the fact that colloidal gold reagents can't penetrate into tissues or cells. The gold particles are too large. These reagents are used for labeling surfaces of sections. If you want to use a gold conjugate for whole or thick tissues sections, prior to embedding, you have to use a nanogold reagent, not colloidal gold.
What do you mean that, "After the tissue is emersed in antibody, we see periodic decoration of the fibrils. How do you visualize this?
Joseph Goodhouse Confocal / EM Core Lab Manager Department of Molecular Biology Princeton University jgoodhose-at-molbio.princeton.edu 609-258-5432 Info and Images at http://www.molbio.princeton.edu/confocal/CF-EM-HOME.html
World Oil's Fourth International Forum and Exhibition on Exploration and Production Management Aug 31-Sept 2, 1998, Marriott Westside Hotel Houston Texas.
World Oil is proud to announce the fourth forum that offers E&P managers an overview of the latest technology available in providing data management solutions.
Today's E&P managers are being confronted with the challenge to gain control of the constantly expanding volumes of E&P information. To stay competitive, cost-effective and timely E&P decisions are necessary. This means accessing the large amounts of data inherent in today's projects and providing that data to your geo-scientists in an efficient and effective manner.
Key features of our 1998 "Forum" program:
*Year 2000 compliance issues and solutions *Real-life case histories *Timely solutions produced with existing data management and software applications *Successful cooperation between operating companies and service organizations *Outsourcing alternatives to reduce fixed cost and allow E&P personnel on finding/producing oil and gas *Determining when older technology should be replaced with new approaches *Deciding how to implement new technology and produce early results *Defining the role of the decision-making manager in information management activities *Maintaining the flow of information over the life span of an E&P asset *Utilizing Geographic Information Systems (GIS) to access and use E&P data
For more information contact Mel Ladin at 1-800-231-6275 or (713) 520-4445. You may also visit our Website for the most current information and register on-line: www.gulfpub.com
To be removed from our data base, type remove and reply to betsy_74-at-hotmail.com or call 904-771-9410.
I just have to react to your message posted on the Microscopy Listserver as a reply to Doug Keene's question. I fully agree with you that limited or lack of penetration into the section interior may very well be the cause for negative results. I had a message from Doug where he indicates that he will be looking into this, using FITC labels for LM first and ultra small colloidal gold labels with silver enhancement for EM if the first approach proves to be successful. In your answer you say that "you have to use a nanogold reagent, not colloidal gold". As the principal inventors of ultra small colloidal gold particles we demonstrated already back in 1988 that conjugates based on ultra small colloidal particles are very well able to penetrate under circumstances where 5 or 10 nm particle based conjugates will not do so. If you like I will be happy to send you the documentation. The reagents you refer to didn't even exist at that time, although undecagold was known and succesfully used for high resolution immunoelectron microscopy without silver enhancement. The colloidal ultra small labels (sold by several companies with a reputation in gold labels) as well as gold clusters may solve the problem.
Jan
============================= Jan Leunissen, Ph.D. AURION ImmunoGold Reagents & Accessories Managing Director Costerweg 5, 6702 AA Wageningen The Netherlands
AutoMatch is a public domain macro written for the public domain software NIH Image. It is described in an article by Swidbert R. Ott, Microscopy Research and Technique 38:335-339(1997). The montage is automated. The article indicates that the software is available via FTP from zippy.nimh.nih.gov/pub/nih-image/contrib/
Mark, try the imtools package from the SDSC group. Specifically, it contains a program alled imstoryboard that allows you to make a montage of many images into a single file ready for printing. You have control over spacing between images, between images and the edges, the background color, etc. Another way would be to use the netpbm package. Using pnmcat one can concatenate multiple images into a single image. As this one works with standard in and output, quite possibly one could simply redirect the output of it straight to the printer, as in:
pnmcat pnmfile1 pnmfile2 ... | lp (or lpr)
Hope this helps.
Jaap
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Tue, 30 Jun 1998 mark_munro-at-bio-rad.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } does anyone know of a graphics package that can take several separate } digital image files and montage them on a single page ready for printing? } } Thanks, } } } Mark Munro }
We have the same software and might be able to help with some questions, at least on the image processing side.
Remember, the imaging part is Noesis Vision's Visilog product rolled into the Oxford suite of products.
If anyone would be interested in discussing either Visilog, or ImQuant in particular, I would be willing to set up a mailing list for discussion. Please contact me directly.
At 03:25 PM 6/30/98 +0000, you wrote: } } Has anyone got some tutorials for this Image software. The manual is } too complicated... } } F. (frank.sarrazit)
For calibrating our Mettler FP82HT hot stage, we have been using the melting points of organic compounds supplied with the Kofler Hotbench (Eichsubstanzen fuer Kofler Heizbank). These are now several years old, and someone has suggested that liquid crystals might be more apporopriate.
I would be glad if anyone can update me as to what is state-of-the-art in calibrating hotstages.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Doug Keene wrote, } } Dear Microscopists: } } I am in a mild state of confusion regarding an immuno-EM } experiment where it seems that the secondary antibody (GAM } 5 nm) does not recognize the primary (mouse IgG) once the } primary is bound to its corresponding antigen in tissue. } We expect that the antigen is present with periodicity on } fibrils in the connective tissue matrix. After the tissue } is emersed in antibody, we see periodic decoration of the } fibrils, which indicates to us that the antibody is bound. } However, secondary antibody does not bind to the tissue. We } are convinced that the secondary conjugate is not } defective, as we use it for other experiments. Has anyone } else experienced a situation where a secondary does not } recognize a primary once the primary is bound to its } corresponding antigen? } } The problem is not that the secondary does not recognize your } primary. It is the fact that colloidal gold reagents can't penetrate } into tissues or cells. The gold particles are too large. These } reagents are used for labeling surfaces of sections. If you want to use } a gold conjugate for whole or thick tissues sections, prior to } embedding, you have to use a nanogold reagent, not colloidal gold. } } What do you mean that, "After the tissue is emersed in } antibody, we see periodic decoration of the fibrils. How do you } visualize this? } } Joseph Goodhouse } Confocal / EM Core Lab Manager } Department of Molecular Biology } Princeton University } jgoodhose-at-molbio.princeton.edu } 609-258-5432 } Info and Images at } http://www.molbio.princeton.edu/confocal/CF-EM-HOME.html
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
We are getting good results with the Polaroid scanner which is really the Microtek scanner in a Polaroid box. It is not as good resolution for 35mm as the Sprintscan, but in some respects it is better in that a whole bunch of frames can be digitized at once. But as for NT compatibility, we have not tried it on any of the NT machines so we don't know.
-------------------------------------------- Michael Cammer email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 http://leper1.ca.aecom.yu.edu/aif/ --------------------------------------------
On Fri, 26 Jun 1998, Joiner Cartwright, Jr. wrote: } } we want to buy a scanner which can be used for both 35 mm and 70 mm EM } } film as well as 81x100 mm plates, probably with different adapters (?).
Our imaging database, ElectroImages, can montage images easily and with great flexibility. ElectroImages also allows the user to place text captions under images. Captions can include title, date, source, extensive descriptions, filename, filepath, and up to 6 user-definable fields. You can also easily print a footer and header to give your page a finished look.
Please contact me off-list and I will send you our trial copy of this program.
Matt Irwin ElectroImage, Inc. 277 Northern Blvd. Suite 101 Great Neck, NY 11021
LEO Electron Microscopy is hiring SEM service people in California. If you are a proven SEM service/support person, and would like to practise your art for LEO in the SF or LA areas, please contact me direct (billneill-at-csi.com)
} Experienced Electron Microscopist } } A prominent cardiac muscle physiology laboratory at the } University of Pennsylvania has an opening for an experienced electron } microscopist. The individual must be proficient in all aspects of } conventional transmission EM from preparation of the samples to } publication } quality photography. Knowledge of some standard biochemical } procedures } including electrophoresis would be helpful but not essential. We are } looking for an individual who is willing and can demonstrate the } ability to } learn and execute new techniques. A B.S. degree is highly desirable } but not } absolutely essential. Salary will be commensurate with experience. } Please } contact Dr. Saul Winegrad. } } E-mail address is bsg-at-mail.med.upenn.edu } } } } }
I am looking for an instruction of sufficient detection of apoptotic cells in paraplast embedded tissue. Is anybody able to point me to a reference with an adequate recipe or can mail me one?
-- Mit freundlichen Gruessen Yours sincerely *************************************************************** * T. J. Filler | * * Westfalian Wilhelms-Univ.| phone:*49 251 83 552-26 Fax: -41 * * Institute of Anatomy | e-Mail: filler-at-uni-muenster.de * * Vesaliusweg 2-4 | D-48149 Muenster Germany * ****** http://medweb.uni-muenster.de/institute/anat ***********
Warren wrote: "Offhand, I would be suspicious of the cooling fan/system. I have seen a number of computers get flaky once the processor overheated. My first experience of that type was after installing a 486-50 overdrive processor in a PS/1. (Nobody told me it needed a cooling fan.) Things come to a halt very quickly once the processor overheats, and it doesn't take all that long to warm up. 5-10 minutes sounds like plenty long enough. I have had the same experience with trying to drive a processor a little to hard, like when I was trying to push my 120 MHz up to 133 MHz. It didn't seem like much of a push, but the manufacturers have already well pushed to the edge so that there is no room for a cooling fan to fail"
But the real truth is that CPUs are rated extremely conservatively. Typically, the maximum speed diminishes as they age, and no responsible manufacturer wants to risk having to replace millions of microprocessors on warranty. That is why there can be considerable ability to "hot rod" CPUs, and still keep them running-for a while. Today's CPUs have the clock speed burned in at the factory, and will be reliable for very long periods of time.
-----Message d'origine----- De : Lucille A. Giannuzzi {lag-at-pegasus.cc.ucf.edu} =C0 : microscopy listserver {microscopy-at-sparc5.microscopy.com} Date : mercredi 1 juillet 1998 05:10 Objet : SF6 upgrades for JEOL TEMs
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Lucille
The new norm about CFC's regulation don't take care of the use you'll mak= e of freon. The important thing is to not let freon going to the atmosphere and you cannot be sure of that during emergency on the gun or on the HT tank. This pollution is too bad and freon will not be available soon. I think your provider has to sell his stock which we will not be abble to u= se later.
I received a message from a coworker asking if I had any information to help you.
I am sending you a web site that deals with old scientific instruments and of course includes microscopes. I purchased two book from them, which might help:
1. Turner, G.L'E. COLLECTING MICROSCOPES $25.00 Christie's International Collectors Series New York: Mayflower Books, 1981 8vo, pp 120; cloth, dj; new copies (out-of-print) with 102 instruments illustrated, many in color
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; AFM Technology Leaders for Environmental / In Vitro AFM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
The Adem was ahead of it's time. It has a six axis stage, fully integrated imaging, and X-ray analysis. The column can accept very large samples, and it has a motor for everything. They were coming out with a FE model when they pulled the plug back in the early 90's. This was due to several factors. The 5500 series 2 X-ray system was nearing the end of it's sales cycle, and the other Microscope vendors did not take too kindly to the competition. This caused sales alliances with the OEM's to collapse. Throw in the fact that Noran was bought and sold twice in that time frame. So the new management felt they had to save the core business. There were still some working units out there when I left Noran 2 years ago.I worked on them briefly and I found them to be difficult to work on. The concept of a fully integrated SEM was sound, however, they made it way too complicated. If Noran could have held on for a while longer they would probably have a good market share. I found it interesting that the chief designer Fred Schamber went to R J Lee and designed the personal SEM. The same concept only on a much simpler scale.
Thu, 2 Jul 1998 09:25:13 +0100 Received: from localhost by spsscsc2.rdg.ac.uk (8.8.5/8.8.5) with SMTP id JAA24200; Thu, 2 Jul 1998 09:25:00 +0100 (BST)
On Tue, 30 Jun 1998 mark_munro-at-bio-rad.com wrote:
} Dear all, } does anyone know of a graphics package that can take several separate } digital image files and montage them on a single page ready for printing?
For a GENERALLY USEFUL package which can do things in a nice way, try downloading a trial version of Corel Xara! 2.0 from:
http://xaraxone.i-us.com/
I have only recently acquired this, so I haven't tried out all its functions, but I have started converting some of my black-and-white TEM images to duotone using this package.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
If you want to impress the kiddies, go ahead and use those liquid crystals. After all, it sounds much fancier that way.
But I certainly wouldn't bother. There is nothing at all wrong with regular melting point standard substances. And viewing in cross polarizers, the melting temperature is always quite clear. I certainly wouldn't buy anything new if I were sure that the existing materials were OK. (If they're not, the melting point is no longer sharp. You can test this in a DSC.)
Was the person suggesting other standards in any chance a salesman wanting to sell them to you?
Cynthia Bennett Hoechst Diafoil
The opinions expressed here are solely my own. Don't blame my employer.
------------------------------------------------------------------------ - Robert Olley wrote:
For calibrating our Mettler FP82HT hot stage, we have been using the melting points of organic compounds supplied with the Kofler Hotbench (Eichsubstanzen fuer Kofler Heizbank). These are now several years old, and someone has suggested that liquid crystals might be more apporopriate.
I would be glad if anyone can update me as to what is state-of-the-art in calibrating hotstages.
+----------------------------------------------------------------------- -+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley |
There will be a meeting of the MSA Standards Committee and ASTM committees E42-96 (US TAG for ISO TC202 Microbeam Analysis) and E42-15 (Electron Probe Microanalysis/Electron Microscopy) on Thursday July 16th at 12:30 PM in Rm. 275-W of the Georgia World Congress Center. For additional information contact john.small-at-nist.gov.
Hi! Does anybody know if there is any problems (eg precipitate within the tissue, etc) if tissue is fixed in 2% osmium tetroxide within a stainless steel container? Thanks Dorota
Everyone was so helpful when I was looking for a large SEM chamber. Can you help again?
I had a call from a science museum in Virginia. They are setting up a display and would like an illustration of the crystallographic structure of aluminum and if possible, a 7079 alloy.
I have at my ready grasp a drawing of graphite, which is close. But museums tend to be very particular and they should have aluminum. They would like the drawing for reference in constructing a model.
If you have such a drawing, please contact Mr. Brian Alfano at balfano-at-smv.mus.va.us
We have funding to buy a new SEM and have at least one application for an = environmental(variable pressure) microscope. Those of you who have = experience with environmental scopes, especially the Hitachi 3500N, please = let me know the good and bad points about your scope. Most of our samples are animal tissue and some of my reading has = suggested that to prevent tissue drying, when working at low vacuum, it is = necessary to use a cold stage. How do you rate the importance of a cold = stage for this type of specimen? =20 Finally, those of you who have a cold substage from Fullam... have you = been satisfied with its performance? =20 Any comments are welcomed. Thanks.
Donna Wagahoff SIU School of Medicine PO Box 19230 Springfield, Il 62794-1220 217-782-0898
Here's a general question that should stir up some comment...
My department has sent out a standard questionaire for us to estimate any problems our lab computers, software, or equipment might have with the year 2000 date problem. Perhaps some of you have already gone through this. I'm not a computer wiz, and naively thought that everything I use would be fine, since I don't do any date calculations like folks in the banking, insurance, etc. businesses do. However, the more I hear about this problem the more confused I get. Is it true that some older CPU's or BIOS programs will not handle the "00" date? So some of our older equipment might not work even though the software we are aware of has little reliance on dates?? Any microscope or software vendors want to comment??
Hope this isn't stirring up needless fears or urban legends, but I have definitely heard some alarmist opinions and would like to consult a more technically aware and level-headed source (i.e. yourselves!).
Thanks, Karen
-- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
I wish to inject ferritin into the hemolymph (blood) of crabs to determine which spaces are blood spaces. (Crab blood has copper-based hemocyanin, rather than hemoglobin).
Sigma carries several varieties of Ferritin, including Apoferritin. Any suggestions as to which one I should use?
Does anyone have a protocol to recommend?
What final concentration should I shoot for in the blood stream?
Thanks to all respondents.
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
A recent breakthrough in environmental microscopy technology is MAC Mode-AFM. This will allow you to do in vitro, time lapse imaging at a nanometer scale resolution.
You can use a 37 C temperature control with flow through liquid cell to change your biological buffers and observe conformational change at the molecular level.
Also you can characterize attractive and repulsive forces between antibody - antigen or other bio agents. A great paper, "Antibody-Antigen Recognition Detected By Ultra sensitive Force Microscopy" will be presented at Protein 98 san Diego by Peter Hinterdofer of the Schindler group from Linz Austria.
George
At 05:55 PM 7/2/98 -0600, Donna Wagahoff wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have experience with e.g. marine samples in ESEM? Or tissue in buffers? I am curious to know whether they are feasible. With samples, such as small planktonic organisms which are living in what is essentially 3% sodium chloride solution, can they be dried off without getting a coating of salt crystals? I know from cryoSEM that once the water is removed, you get the covering of salts. I imagine that some items might withstand a quick rinse in fresh water, but not everything.
Karen, Most old PCs (but not Macs or most Unix) will not transfer the date correctly to the year 2000 although many will handle dates after Jan 1, 2000 if told explicitly (i.e. with 4 figure years). You can get free test software for PCs off the www. I found one from National Software Testing Laboratories (NSTL) that seemed pretty useful. If your old PC will not do the transition automatically you can either roll the date forward manually (if it can do this), use a false date (such as 1990) but obviously this could be misleading in some cases, or scrap the PC and buy a new one.
For the most of our PCs that control lab equipment in our group I know that they will not roll the date over automatically, so I am likely to keep all the data backed up, wait till the end of 1999 and see what happens. If one dies for some reason then we can always replace it with another cheap PC.
Hope this helps
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
This is a multi-part message in MIME format. --------------99D16DAA7889B787EDE9016F Content-Type: text/plain; charset=big5 Content-Transfer-Encoding: 7bit
A postdoctoral position in scanning tunneling microscope investigation on the interfacial reactions of metal thin films on silicon is available at the
IC Thin Film Lab. Department of Materials Science and Engineering National Tsing Hua University Hsinchu, Taiwan, Republic of China
The position is for one year and renewable for two more years. Interested persons should have experiences in UHV-STM. Please contact Professor Lih J. Chen directly either by e-mail or by fax at 886-3-5718328.
--------------99D16DAA7889B787EDE9016F Content-Type: text/x-vcard; charset=big5; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Lih J. Chen Content-Disposition: attachment; filename="vcard.vcf"
begin: vcard fn: Lih J. Chen n: Chen;Lih J. org: Department of Materials Science and Engineering, National Tsing Hua University adr: Department of Materials Science, National Tsing Hua University;;;Hsinchu;Taiwan;300;Republic of China email;internet: ljchen-at-mse.nthu.edu.tw title: Professor tel;work: 886-3-5731166 tel;fax: 886-3-5718328 x-mozilla-cpt: ;0 x-mozilla-html: FALSE version: 2.1 end: vcard
The question has been posed whether cells grown in culture or cells from sea water can be imaged using ESEM. As a general principle I have found it possible to image cells in culture medium - crystallisation upon drying. It is possible to image cells from 0.9% saline and in such cases the NaCl crystals are widely dispersed and mostly do not interfere with imaging (sod's law not withstanding!) I can try 3% saline to see what happens and will let you know.
Another question needs to be addressed however. What are you trying to see? I have much success by viewing glutaraldehyde fixed samples washed in water and imaged with ESEM. The cells remain hydrated are much more robust (resistant to accidental drying, withstanding beam damage). On most occasions I have not seen structural differences between fresh hydrated and fixed hydrated. It is the drying which causes changes not fixation.
As with all SEM low kV should be used (hard work below 5KV unless you have Brendan Griffin detector modification). Check the pressure temperature curve for water vapour pressure but 3 - 4 degrees C with a pressure of 5 - 6 torr should keep specimens hydrated.
Good luck Let me know if I can help further.
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
If you are attending the MSA Meeting in Atlanta, Mike Boykin and I would like to invite you to a gay social on Monday, July 13th at 8:00pm. The party will be held at Mike's home in midtown Atlanta. Directions to Mike's house will be available via email or at the Information and Hospitality Booth. Also, copies of Southern Voice (a gay weekly newspaper) will be available. If you are planning on attending the social please RSVP to me. See y'all there, Beth Richardson
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
For those interested, I thought I would summarize the comments received regarding my last querry to the listserver, which was:
Dear Microscopists,
I am in a mild state of confusion regarding an immuno-EM experiment where it seems that the secondary antibody (GAM 5 nm) does not recognize the primary (mouse IgG) once the primary is bound to its corresponding antigen in tissue. We expect that the antigen is present with periodicity on fibrils in the connective tissue matrix. After the tissue is emersed in antibody, we see periodic decoration of the fibrils, which indicates to us that the antibody is bound.
However, secondary antibody does not bind to the tissue. We are convinced that the secondary conjugate is not defective, as we use it for other experiments. Has anyone else experienced a situation where a secondary does not recognize a primary once the primary is bound to its corresponding antigen?
Responses:
Several responses pointed out that secondary antibody conjugates are sub-type specific, meaning that if my primary was really an IgM and not an IgG, that a GAM IgG would not recogize the IgM. In my case, I know that the primary is an IgG.
Several responses suggested that I might not reasonably expect primary and secondaries to penetrate into the connective tissue matrix, that it might be to dense. Therefore I should consider a surface labeling procedure during which the antigen is exposed via sectioning. Unfortunately, this antibody will not work in surface labeling procedures, even though I have tried many different fixation/embedding techniques. Also, we have had years of success diffusing antibody into the CT space, including antibodies to other components of this same microfibril. However, as pointed out by another response, the gold particule we are using (5 nm) may be to big, and steric hindrance might be an issue. The suggestion is to try LM first, with a smaller FITC conjugate, then if positive use a smaller (1 nm) secondary for EM. This is the procedure we are about to try.
Many people still call this meeting MSA, but I do believe that the meeting is sponsored by the Microbeam Analysis Society and the Microscopy Society of America and that is why it is known as Microcopy and Microanalysis 98. There seems to be a general lack of understanding that the meeting is a joint affair of the two socities.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This sounds more probable with Environmental SPM imaging in the 3% sodium chloride solution / buffer. There is not need to dry it of and you can image in vitro.
George
At 08:16 AM 7/3/98 +0100, Keith Ryan wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Donald, } } I wish to inject ferritin into the hemolymph (blood) of crabs to determine } which spaces are blood spaces. (Crab blood has copper-based hemocyanin, } rather than hemoglobin). } } Sigma carries several varieties of Ferritin, including Apoferritin. Any } suggestions as to which one I should use? } I think the apoferritin is the protein without the iron; if so, it would be a very poor tracer.
} Does anyone have a protocol to recommend? } If you can map the iron, you can see where it appears and where it is co-localized with copper. If you do not have element-mapping capability, the problem is much more difficult. In the latter case, some gold-labelled anti-ferritin (or forgetting the ferritin altogether, gold-labelled anti- hemocyanin) could work.
} What final concentration should I shoot for in the blood stream? } For EDS you would need a reasonable fraction of 1% iron; for WDS you need much less--I'll let the WDS experts tackle this one. Good luck. Yours, Bill Tivol
MSA's middle school microscpoy manual has JUST been published! Here's a brief description (taken from the Project MICRO bibliography):
Brady, S. and Willard, C. 1998 Microscopic Explorations 158pp, paperback, 8.5x11". ISBN 0-924886-00-5 Lawrence Hall of Science, University of California, Berkeley, CA 94720-5200; 510-642-7771 or 7262, gems-at-uclink.berkeley.edu. A collaboration between the Microscopy Society of America and the LHS has produced an outstanding Great Explorations in Math and Science (GEMS) guide. It's written in "festival" format, with a dozen explorations that can be presented simultaneously to circulating groups of students. There is a rich assortment of supplemental information on microscopes and how to buy them, curriculum extensions, further reading, and sources of help. The units are more classic than unique; subjects include crystals, color printing, fingerprints, pond water, brine shrimp, etc. Its uniqueness lies in the carefully written "inquiry science" presentation of those topics and the thorough classroom testing of content that a GEMS guide receives. It will work well in any classroom; teachers aren't expected to have special skills. Information on other GEMS guides (there are over 50!) is available at www.lhs.berkeley.edu/GEMS. Grades 4-8.
The retail price is $21.00, plus $4.00 shipping (in the U.S.); MSA members may request a 15% discount. A limited number of copies will be available at the Project MICRO booth in Atlanta. See y'all there!
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I've put some of the material from our LASER diffraction workshop on the web*, so as to share the fun. This began as a 2.5 hour workshop for gifted high school students participating in a six week program (the "Engleman Institute") here at UM-StL. A bit of thought, however, has suggested that even some of our diffraction experts might enjoy both the subjects, and the competition.
I would encourage microscopists to check out there computers with regard to the Y2K problem. However, I expect that the impact will be minimal, if not trivial, in practically all cases. We don't have the same sort of issues to deal with as the financial and insurance people.
I just checked out our computers using the test program from NSTL described by Ian below (the software is available from http://www.nstl.com/html/y2k_faq.html). It seems to do a good job of finding the problems, if any.
Our new computers (Pentiums and newer) checked out okay.
Our two remaining 486 machines reported minimal problems. The date would not correctly rollover on 1-1-2000. If the computer was powered off and on after 1-1-2000, the system would come up with a date of 1980. However, if the DOS DATE command was given on or after 1-1-2000 and the system restarted, it would come up with a correct date. There is a "century bit" in the BIOS info somewhere that will stay set once set, but does not appear to roll over correctly on these old machines.
Therefore, we can simply use the DOS DATE command after 1-1-2000 and reboot. Or I found that a utility form RightTime (www.RightTime.com) called YEAR2000.COM will fix the BIOS so that the computer passes the NSTL test. I do not know if it needs to be kept loaded after 1-1-2000. I would definitely want to check before I removed it.
The Year2000.txt file that comes with the RightTime utility does a very good job describing what the technical issues are for those who care to look deeper.
BIOS fixes are available for many computers to fix the problem. However, they may not be woth the hassle unless they are needed to fix another problem.
As far as application problems, I cannot imagine much happening in microscopy programs. If programs use only a two character year code, the code should still get transferred correctly. The problems will probably crop up when sorting or searching according to date. I have noticed the following behavior with some of our programs.
Access 7.0 (for 95) - Entering a value of 1/1/00 will be interpreted as 1900. You can and must enter 2000 (four digits) as the year for it to register as the next century. Otherwise 00 and 01 will precede 98 and 99 when sorted in ascending order.
Excel 95 - Entering 2-digit dates 00 through 19 is treated as 2001 to 2019. (You can check by formating cells to show the full 4-digit date.) 2-digit dates from 20 through 99 are treated as 1920 through 1999. In other words, their two digit century runs from 1920 through 2019.
Paradox database (as used by Quartz PCI, and others) - A search for dates after 1/1/30 return no files, while a search for files before 12/31/29 returnes all files. Their two digit century apparently is defined to cover the years 1930 through 2029.
To Summarize: A - Nothing broke in a real bad way. Even if something did, I suppose that 1999 could be extended artifically by backdating the computer until a suitable fix was found.
B - Many 486 and earlier computers will probably need some attention, but can probably get by with manual use of the DATE command after 1/1/2000. BIOS fixes are available for many computers if someone wants to go that route.
C - The data may have the correct two digit year code, but you may have to be wary how you search or sort the data. As long as manufacturers have written their code so that the two digit century encompasses the full range of our data collection, we should be all right. And since I don't have any data files collected between 1920 and 1929, I think I can safely assume that two digit codes representing the years 1930 through 2019 for my files will be treated correctly.
Hoping this helps, Warren Straszheim
At 10:26 AM 7/3/98 +0100, Ian Mc Laren wrote: } Karen, } Most old PCs (but not Macs or most Unix) will not transfer the date } correctly to the year 2000 although many will handle dates after Jan 1, } 2000 if told explicitly (i.e. with 4 figure years). You can get free test } software for PCs off the www. I found one from National Software Testing } Laboratories (NSTL) that seemed pretty useful. If your old PC will not do } the transition automatically you can either roll the date forward manually } (if it can do this), use a false date (such as 1990) but obviously this } could be misleading in some cases, or scrap the PC and buy a new one. } } For the most of our PCs that control lab equipment in our group I know that } they will not roll the date over automatically, so I am likely to keep all } the data backed up, wait till the end of 1999 and see what happens. If one } dies for some reason then we can always replace it with another cheap PC.
I am looking for PTFE tubing to fit a Hexland cryostage. Does anyone know who carries parts for this unit?
Thanks, Carl
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
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Hi Donna, Hope all goes well with you. Haven't talked to you ikn a long time. ESEM There is actually only one true environmental scanning EM and that is the = Philips Electroscan. The advantage is that samples are kept humid and do = not dry out. Any other "Vairiable pressure" SEM dries out the sample = with only short times to look at it. SO, as everything else, it depends = on what you want to do with the scope. If you are really looking at wet = samples, then the true environmental SEM seems appropriate. If that = isn't of interest then the others are cheaper. My two cents, Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
We have funding to buy a new SEM and have at least one application for an = environmental(variable pressure) microscope. Those of you who have = experience with environmental scopes, especially the Hitachi 3500N, please= let me know the good and bad points about your scope. Most of our samples are animal tissue and some of my reading has = suggested that to prevent tissue drying, when working at low vacuum, it = is necessary to use a cold stage. How do you rate the importance of a = cold stage for this type of specimen? Finally, those of you who have a cold substage from Fullam... have you = been satisfied with its performance? Any comments are welcomed. Thanks.
Donna Wagahoff SIU School of Medicine PO Box 19230 Springfield, Il 62794-1220 217-782-0898
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Dear List,
We recently purchased a microwave system for tissue fixation/embedding. Unfortunately, the instruction book that we ordered on it's use did not come and we now find out that it is no longer available (through Ted Pella). Does anyone have any information on procotols or know of any good reference books on microwave fixation and embedding? Any information that will get us going would be greatly appreciated.
William R.McManus Supervisor Electron Microscope Facility Department of Biology Logan UT 84322-5305 billEMac-at-cc.usu.edu
Looking for a new/used copy of: THE STUDY OF PLANT STRUCTURE: PRICIPLES AND SELECTED METHODS. 1981. O'Brien, T.P. and McCully, M.E. Termarcarphi Pty. Ltd. Melbourne, Australia.
Also, looking for steel (non-disposible, and glass blades for a Spencer 820 American Optical Rotary Microtome.
Thank you in advance Please reply via email: crow-at-aloha.net
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear List, } } We recently purchased a microwave system for tissue fixation/embedding. } Unfortunately, the instruction book that we ordered on it's use did not } come and we now find out that it is no longer available (through Ted } Pella). Does anyone have any information on procotols or know of any good } reference books on microwave fixation and embedding? Any information that } will get us going would be greatly appreciated. } } William R.McManus } Supervisor } Electron Microscope Facility } Department of Biology } Logan UT 84322-5305 } billEMac-at-cc.usu.edu } } } Bill, Here is a reference which may be of some help. Ted Pella equipment was used throughout: J Vet Diagn Invest 9:61-67 (1997) "Four-hour processing of clinical/diagnostic specimens for electron microscopy using microwave technique."
The Microwave Tool Book--A practical guide for microscopists by Gary Login and Ann Dvorak ISBN 0-9642675-0-0
Microwave Cookbook for Microscopists--Art and Science of Visualization by L. P. Kok and M. E. Boon ISBN 90-71421-20-1
Rick Giberson at Ted Pella should be able to give you some starting point protocols.
Patty Jansma Tel:520-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
On Mon, 6 Jul 1998 billemac-at-cc.usu.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear List, } } We recently purchased a microwave system for tissue fixation/embedding. } Unfortunately, the instruction book that we ordered on it's use did not } come and we now find out that it is no longer available (through Ted } Pella). Does anyone have any information on procotols or know of any good } reference books on microwave fixation and embedding? Any information that } will get us going would be greatly appreciated. } } William R.McManus } Supervisor } Electron Microscope Facility } Department of Biology } Logan UT 84322-5305 } billEMac-at-cc.usu.edu } }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Greetings, } } I am looking for PTFE tubing to fit a Hexland cryostage. Does anyone know } who carries parts for this unit? } } Thanks, } Carl
Hi Carl, Hexland are now part of Oxford Instruments, Research Instruments, Tubney Woods, Abingdon, Oxford, OX13 5QS. UK. +44 1865 393200. Or try your local agent. If you do not know a local contact e-mail Judith Brock on judith.brock-at-oxinst.co.uk.
Regards, Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
We are looking for flat molds for embedding in acrylic resins which could be hermitically closed. Can one out of you send me the name of the companies which sells them.
Thanks a lot
Marc
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
We are currently trying to establish capability in our lab to look at die cross-sections. In previous work we mounted them in epoxy or acrylic and coated the samples, but I'd like to be able to do this without mounting and coating. I've been told that there are simple polishing fixtures one can by to do this, but I can't seem to find where to buy them. Perhaps someone out there could suggest a source? Thanks,
Janet Rice MCC Senior Member Technical Staaff rice-at-mcc.com 512-338-3266
} We are currently trying to establish capability in our lab to look at di= e } cross-sections. In previous work we mounted them in epoxy or acrylic an= d } coated the samples, but I'd like to be able to do this without mounting and } coating. I've been told that there are simple polishing fixtures one ca= n } by to do this, but I can't seem to find where to buy them. Perhaps someone } out there could suggest a source? Thanks
--------- BUEHLER supplies the MICRO-PRECISE(TM) line of IC cross-sectioning equipment including the Tripoint Polisher for SEM/TEM sectioning. We als= o supply a semi-automated system that might be appropriate for your application as well. For more information, please contact me directly.
Scott D. Holt BUEHLER LTD. PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-6500 http://www.buehlerltd.com
The technique you are describing is very well suited to our Tripod Polish= er and is most likely the tool you have heard about. The Tripod Polisher is=
ideal for preparing a cross section of a specific device and can be used = to prepare both SEM and TEM cross sections. The SEM cross section is actual= ly the "easy" part of the process and is referred to as our "first side polish" when preparing a TEM cross section. The Tripod Polisher is easy = to use and we have developed a large database of technical reports on its us= e. The Tripod Polishing technique and the tool were actually developed at I= BM East Fishkill and have been used in cross sectioning for many years.
Another alternative is our BiPod Polisher which is a simpler version of t= he Tripod Polisher and is designed primarily for SEM cross sectioning. The BiPod polisher can be mounted on our polishing machines for semi-automati= c cross-sectioning. I'll be pleased to send you additional information on any of these products if they sound of interest to you. You can also get=
some of the information off of our web site at http://www.southbaytech.co= m.
If you plan to be at MSA next week in Atlanta, you can get a thorough demonstration of the technique at our booth (#433). =
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Janet Rice } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
We are currently trying to establish capability in our lab to look at die=
cross-sections. In previous work we mounted them in epoxy or acrylic and=
coated the samples, but I'd like to be able to do this without mounting a= nd coating. I've been told that there are simple polishing fixtures one can=
by to do this, but I can't seem to find where to buy them. Perhaps someo= ne out there could suggest a source? Thanks,
} We are looking for flat molds for embedding in acrylic resins which could } be hermitically closed. Can one out of you send me the name of the } companies which sells them.
} Marc -
The Ted Pella company has one. It's teflon, and should be sealed for polymerization with Thermanox cover slips. I know it works, because it was designed by Doug Davis, a tech in my (pre-retirement) lab at U.C.Berkeley.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Is anyone currently doing the Kleinschmidt procedure for visualization of DNA with TEM? Dave Long at Montana State University needs to cooperate with a microscopy lab to have some DNA samples shadowed at low angle with the Kleinschmidt procedure. Either contact him by phone (406-994-5239) or me by phone (801-378-2451) or e-mail. Thank you. W. M. Hess.
Hi, I was asked to post this information from Cynthia Goldsmith for all M&M '98 attendees.
Taking MARTA (Metro Atlanta Rapid Transport Authority) from Hartsfield International airport to the downtown hotels.
} The MARTA rail can be found at the end of the baggage claim areas. Take } the MARTA Northbound train to the Peachtree Center station (N1), then } follow the signs to either the Marriott Marquis (through the Peachtree } Center Food Court and across a ramp) or the Westin Peachtree Plaza } (across Peachtree Street). } } You could also mention (as Sandy has in the information brochure) that } the rail lines run North-South and East-West, with the transfer point at } the Five Points Station. The GWCC (Georgia World Congress Center) is on the } } E-W Line, at the W1 station (Omni/Dome/GWCC).
see y'all soon, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
We have a couple of older Kevex Delta (III, IV) EDS units based on DEC PDP-1173's running RT-11 ver. 4.05. The units will not rollover to from Dec 31, 1999 to Jan 1, 2000, instead giving a date error. The RT-11 ver. 4.05 operating system is not year 2000 compliant. Once the Kevex software is up and running it will accept the year 2000 dates and run fine, however if you reboot the system the Kevex software takes its date from RT-11 which has to be pre-2000 in order for it to boot. What this means is that if we want to have the correct date on our printouts we'll have to change the dates both in RT-11 and the Kevex software every time we use the system. Currently this is the only date related problem we've noticed with the systems. Of course, we could upgrade to a PC based system or the RT-11operating system and DEC PDP-1173's could be upgraded. Both are now owned and supported by a company called Mentec (http://www.mentec.com/) and the new version of RT-11 (ver. 4.7) is y2000 compliant. An upgrade to ver. 4.7 would cost in the neighborhood of $2k. Mentec has indicated to me that although the new version is y2000 compliant, there are other changes and without input from the developers of the overlying Kevex software they can't guarantee that the Kevex software would run correctly. The PDP-1173 could be upgraded (bournoulli drives are no longer available) with SCSI cards and compatable peripherals and in fact would have to be since Mentec no longer supplys software upgrades on bournoulli's. For now, our strategy will probably be to change dates and hope for an upgrade to PC based systems.
kszaruba-at-MMM.COM wrote:
} Here's a general question that should stir up some comment... } } My department has sent out a standard questionaire for us to estimate } any problems our lab computers, software, or equipment might have with } the year 2000 date problem. Perhaps some of you have already gone } through this. I'm not a computer wiz, and naively thought that } everything I use would be fine, since I don't do any date calculations } like folks in the banking, insurance, etc. businesses do. However, the } more I hear about this problem the more confused I get. Is it true that } some older CPU's or BIOS programs will not handle the "00" date? So } some of our older equipment might not work even though the software we } are aware of has little reliance on dates?? Any microscope or software } vendors want to comment?? } } Hope this isn't stirring up needless fears or urban legends, but I have } definitely heard some alarmist opinions and would like to consult a more } technically aware and level-headed source (i.e. yourselves!). } } Thanks, } Karen } } -- } Karen Zaruba, kszaruba-at-mmm.com } BioMaterials Technology Center } 3M Center Bldg. 270-1S-01 } St. Paul, MN 55144 } } *The opinions above are my own, not necessarily my employer's*
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Our Hitachi 3200H SEM came with a Magneto-Optic drive for storing images. When we transfer these to our PC however, the images show up without any of the labels (Mag. KeV, etc.). Has any one experienced this and does any one know how to correct this ?
Thanks to all who replied to this question. I should have said that the specimens, in the first instance, would be barnacle larvae. These are quite tough compared to a lot of other specimens that we have handled over the years. The study is to do with the question of settlement onto substrates.
The ideal would be to look at specimens with ESEM and then to allow them to grow on - do you think this is possible or would ESEM/radiation dose curtail normal development after exposure to the beam?
In this age of digital imaging, is a film recorder still useful? The last presentation I went to was all PowerPoint 'slides' but shown directly from a computer to a video projector, no film to be seen.
I am trying to decide if we should get one, to transfer digital images back to film and/or to make slides for presentations etc. If you have some opinions or experience with these things, could you let me know.
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu **Area code changing to 831 as of 7/11/98**
Jordi writes ... } } Hi ! } } Our Hitachi 3200H SEM came with a Magneto-Optic drive for storing } images. When we transfer these to our PC however, the } images show up without any of the labels (Mag. KeV, etc.). } Has any one experienced this ...
Yes ... on a different system ... but at least my JEOL system allows an "export" option which "burns" the info you want into the TIF (... assuming it is TIF files you are archiving ...). I respond because we actually prefer that JEOL doesn't do this ... as we prefer to annotate with better software (e.g., Photoshop). If we've lost track of our notes and the associated mag with each image ... we've realized we can load the image into Windows' Wordpad and search for the mag in the TIF header ... we can then pick an appropriate micron bar dimension from a table associated with micron/pixel for each mag ...
Also ... you might ask Photoshop to show you the TIF header info ... you might be able to find the info you're after if your software adhered to TIF's standard comment fields whereas JEOL didn't.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
In response to Donald Lovett who wanted to know about using ferritin as a tracer for the crab. I did a study several years ago using ferritin mixed with india ink as a tracer to study circulation in the squid Loligo. The india ink helped us see when the solution was in the circulatory system. Mix 5 mls of 2X artificial sea water with 5 mls concentrated ferritin (Calbiochem) and 2 drops of india ink (sonicate the ink for 15 mins to break up the chunks). You need to spin down the ferritin for 30 mins -at- 30K to concentrate the ferritin. Then perfuse the animal-it might be hard in a crab-for 30 mins -at- room temperature. Then submerge into a high osmolarity fix (ie 2.5% glut in 0.1M cacodylate buffer with 0.8M sucrose- 1500 mosm). It worked really well. We also tried lanthanum and HRP. The combo of ink and ferritn was the best and easy to see in the TEM. Good luck! JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94022
Jon, Film recorders, as you are aware, are quite expensive ($10K for a good one). For the same amount of money you could buy the computer and projector to show them with. It's tempting, for sure. I am dithering with this same question myself. The only difficulty with going digital is the complete reliance on the technology: what happens if your computer crashes, the digital projector breaks, or both are incompatible. In getting ready for the MSA meeting (right now, in fact), I opted to do the textual slides in PP but to produce conventional slides for the non-textual material. Regular slides are always going to be superior (as is the photographic medium) and everyone has a conventional projector. Now, if you are pressed for time and have access to the technology, then digital is wonderful. If you have to make the purchase now, then go digital unless you really need the resolution.
} In this age of digital imaging, is a film recorder still useful? The last } presentation I went to was all PowerPoint 'slides' but shown directly from } a computer to a video projector, no film to be seen. } } I am trying to decide if we should get one, to transfer digital images back } to film and/or to make slides for presentations etc. If you have some } opinions or experience with these things, could you let me know.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Jon, we use and will continue to use a film recorder. In our case a Lasergraphics machine. Our lab is entirely digital and to be honest rapid computer based Powerpoint presentations are easily produced and often used. However, when it counts (ie: in public, to your peers) nothing is as good as an excellent high resolution 35mm slide. Whether it is to show illusive immunogold particles on cryosections "I am not sure whether you can see this at the back" or the minutae of confocal or decon image stacks there is NO substitute for a decent film recorder used to make images from high resolution digital images.
Simon C. Watkins Ph.D. MRC Path Associate Professor, Director CBI University of Pittsburgh
Allied High Tech specializes in Materiallographic, SEM and TEM sample preparation. We have the necessary tools, techniques, and supplies to ensure the cross-section you achieve is excellent. Allied also has a new polishing system with a micro-positioning head and polisher for doing precision cross sections, the Multi-Prep and TechPrep 8. If a micro-precision head is not what you want, we also have a easy to use hand tool. The procedures and techniques we have developed work very well. We also have a new Diamond Lapping film brochure containing a pictorial guide illustrating the results that can be achieved with Allied's Diamond Lapping Film.
If you are attending the Microscopy show in Atlanta our booth number is 548.
Gary Liechty is your representative in Texas, but any of the staff at Allied would be happy to assist you with any specific question you may have. Allied's goal is total customer satisfaction and the entire staff is committed to it.
For more information you may contact us at 800-675-1118 and visit our web site at: http://www.alliedhightech.com
We look forward to serving you.
Sincerely,
Edward Hirsch
At 09:06 AM 7/7/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
************************************************* Edward A. Hirsch Product Application Specialist Allied High Tech 2376 East Pacifica Place Rancho Dominguez, CA 90220 ph: (800)675-1118 x245 fx: (310)762-6808 http://www.alliedhightech.com *************************************************
Is there an independent technican and/or dose any one on this list know an independent technican they could recommend to dissamble a Philips 300 TEM, located in Montreal, Canada?
} In this age of digital imaging, is a film recorder still useful? The last } presentation I went to was all PowerPoint 'slides' but shown directly from } a computer to a video projector, no film to be seen.
Joe: A very good question. I have been on both sides of this issue. Below are deciding factors in terms of which format I use. 1) How solid is the technology at the point where the lecture is to be given? The concerns here: I have Mac, they always use PC's, the right cable is not there. I made my presentation on Office 98 under Mac OS8.1 and they have System 7.1 loaded with PowerPoint 3. The mismatches can be endless and much harder to resolve when compared to changing a bulb in a slide projector. 2) Is the PowerPoint presentation better than using the slides? Is animation a key feature of the presentation? Or sound? Are the flying bullets better than the sharpness of the slide? How important is screen lumens? Bright slides work better than dim PowerPoint. It is hard to beat 2000 plus lines of slide resolution when compared to 640 by 480 pixels. They have a video projector with a bulb 500 hours on the otherside of dead. 3) How important is it to fix the presentation on the fly? You sized your slides to work in a 30 foot long room and you find yourself in a 100 foot hall. You can not resize 35 mm slides two hours before the lecture. Of course you can extensively rearrange a lecture in an airplane seat; but you sure can't do that with some 35 mm slides. 4) Can your presentation work best with twin screens: two images at a time? This is usually a snap with 35 mm slides but it is still pretty rough with PowerPoint. 5) One must always keep in mind the audience. Recently I saw a really slick computer controlled presentation. Some in the audience were put off by the excellent presentation because it was too slick. An analysis might help here: it was a research lecture being used to strut one's wares for a tenure-track teaching position. The students in the audience thought it was great. Older faculty who did not use computer media thought it was needlessly glitzy. Media proficient faculty were more focused on how the presentation was assembled rather than the message. Predictable but it still caught me off guard.
Finally back to your question. I use both and probably will continue to do so for the next several years. For really important lectures on the road, I have 35 mm slides for backup. The bottom line is, however, which format will the audience respond to best.
Blystone in Texas
Robert V. Blystone, Ph.D. {RBLYSTON-at-Trinity.edu} Professor of Biology Trinity University San Antonio, Texas 78212 210.736-7243 210.736-7229 FAX
I am trying to respond to a message from Reggie Beijer of the Netherlands Forensic Sciences Laboratory. Unfortunately, I have no contact information. Can anyone provide me with an e-mail or FAX address? Please do not reply to the ListServer. Thank you for your assistance (TIA).
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
The Laboratoire d=92Analyse des Materiaux (LAM) of the Centre de Recherche Public-Centre Universitaire of Luxembourg has immediate opening, on a fixed term basis, for a maintenance engineer
The LAM facility includes 1 SEM, 1 TEM, 2 D-SIMS (CAMECA IMS-4f, CAMECA IMS-LAM), 1 S-SIMS (CAMECA/Ion Tof SIMS III) and one machine under development optimized for MCs+ clusters. The machines in operation are used to perform analysis for industries or by students preparing their PhDs.
The maintenance engineer will take care of all instruments existing at LAM (trouble shooting on electronics, maintenance of the physics).
Candidates should have a degree in electronics (baccalaureat plus two to three years), a first experience in maintenance of UHV equipment and be fluent in English. Interviews will take place at LAM, no funding will be provided for overseas trips.
Please send a resume and a list of references to:=20 CRP-CU 162a, avenue de la Fa=EFencerie L-1511 Luxembourg
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu
We just got a film recorder here and I definitely think that they are worth it: (1) good 35mm film recorders record 4,000 lines of resolution (resolution up to a maximum of 4096 horizontal by 2732 vertical lines, or 4096 x 2732) , which generally excedes the resolution of most 35mm films (looking under a LM I can't see the scan lines on 100ASA/ISA film - NO pixelation), (2) the HIGHEST resolution computer projection systems I have seen are 1024 x 860 - for text these are "o.k." but we work so hard to get excellent microscopic images and do we want to loose it all? (Besides a good film recorder costs the same or less than the best projection system, and you don't have to drag the thing with you to insure you have the best images you can) (3) many MANY places don't have good quality projector systems, but almost every where has a slide projector, (4) images take a LOT of disk storage space, so the image presentations don't fit on a diskette - you really need to burn a CD to take along to the meeting (can you guareentee they'll have a Zip drive? Maybe?), (5) Big image files are slow on all but the best computers systems, (6) which system to design the program for? Mac? PC? (7) want to use your notebook computer? Is it upto the task? do you want to play driver games the hour before your talk? (8) For B&W images the best quality I've seen are reproductions on true B&W media (as many have said this holds ture for printers as well), what I do is record the B&W images in negative to standard B&W film (i.e. T-max 100), develop in my darkroom (20-30mins) dry and mount - beautiful positive B&W slides! (Warning: look carefully, apparently negative film is much more senesitive than slide films so you have to get a good film recorder for this.
The one thing I haven't tried yet is taking true digital images back through film, back to a photographic enlargment, and comparing this to a dye-sub print.
O.k., so that's my two cents.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
Look for the "Microwave Cookbook of Pathology. The art of microscopic visualization". by Mathilde Boon and L.P. Kok. Coulomb Press Leyden, Leiden, The Netherlands. ISBN 90-71421-10-4 bound. I have the second revised edition from 1988, but probably there is a newer one available now.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } On Mon, 6 Jul 1998 billemac-at-cc.usu.edu wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear List, } } } } We recently purchased a microwave system for tissue fixation/embedding. } } Unfortunately, the instruction book that we ordered on it's use did not } } come and we now find out that it is no longer available (through Ted } } Pella). Does anyone have any information on procotols or know of any good } } reference books on microwave fixation and embedding? Any information that } } will get us going would be greatly appreciated. } } } } William R.McManus } } Supervisor } } Electron Microscope Facility } } Department of Biology } } Logan UT 84322-5305 } } billEMac-at-cc.usu.edu } } } } } } }
Bill, }
Here is a reference which may be of some help. Ted Pella } equipment was used throughout: J Vet Diagn Invest 9:61-67 (1997) } "Four-hour processing of clinical/diagnostic specimens for electron } microscopy using microwave technique." } } Drs. Marcel Paques Wageningen Centre for Food Sciences p.a. NIZO Food Research Kernhemseweg 2, 6718 ZB Ede P.O.Box 20, 6710 BA Ede The Netherlands Phone: (31) 318-659690 Fax: (31) 318-650400 e-mail: paques-at-nizo.nl
Jon, "Yes" everyone is going to PowerPoint slides but" no" not everyone has a powerbook that they can take to meetings, courses, etc. Having made countless slides using PowerPoint and a digital slide maker, I don't know how we would manage without both at this point.
Example, I recently returned from a 3 week trip to China. Both my husband and I gave talks at 2 major Universityies on what was otherwise a vacation trip. There was no way we were going to drag a powerbook along. Even if we had, the University lecture halls were not equipped with digital projectors or LCD panels for projection of the PowerPoint slides directly from the computer program.
We have been in countless other situations where we wanted to make informal presentations in rooms which were not equipped with digital projectors such as when our extension educators go out to give talks to local groups. It is easy to come up with a 35mm slide projector, set it up and in no time be able to informally present data in the form of slides but more difficult to find suitable digital projectors, etc. for multiple people from one department to use at the same time in different parts of the state.
It is also very easy to tailor a presentation in just a few minutes by having an assortment of slides and picking and chosing for the audience at hand....much faster and easier that going through multiple PowerPoint files to find the slide desired and then copying and rearranging into a new presentation.
Debby Sherman +++++++
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------
Dear List:
In this age of digital imaging, is a film recorder still useful? The last presentation I went to was all PowerPoint 'slides' but shown directly from a computer to a video projector, no film to be seen.
I am trying to decide if we should get one, to transfer digital images back to film and/or to make slides for presentations etc. If you have some opinions or experience with these things, could you let me know.
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu **Area code changing to 831 as of 7/11/98**
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Would you please recommend a short course for Light Element X-Ray Analysis
I have missed Lehigh's class that took place in June of '98.
Thank you in advance.
Darlene Peterson Harvey Eltech Research Center Fairport Harbor, Ohio
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We will be setting up interviews for candidates for the Fall teaching = position at San Joaquin Delta College for electron microscopy. Course = details are below. If you are interested in teaching all 4 courses or 2 = of them, please contact me in Atlanta at the Microscopy and Microanalysis = Meeting July 12-16, 1998. I will be staying at the Marriott Marquis = Hotel. You can leave a message there or on the meeting bulletin board. = A message left at the hotel might be faster. I will be coming into = Atlanta Saturday July 11 and leaving Thursday evening, July 16, 1998.
Please bring your active resume with you. I will have official SJDC = applications which you can fill out. You will of course need names, = addresses and dates of past employers and names, addresses, and phone = numbers for recommendations and typical application type information. We = can fax them to our Human Resources Dept, and then, if appropriate, set = up a conference interview with the search committee from Atlanta. Please = contact me as early as possible in the week. We are up against an = extremely tight deadline.
Thanks
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
URGENT: ADJUNCT FACULTY NEEDED FALL Semester 1998 (Aug. 12 - Dec 18, 1998)
A San Joaquin Delta College Faculty member of the Microscopy Technology = Center is planning a Sabbatical Leave for the Fall '98 semester. San = Joaquin Delta Community College is currently searching for an EM = Instructor Adjunct / Temporary One Semester Contract for Fall 98 (Aug. 12 = - Dec 18, 1998).
MINIMUM QUALIFICATIONS: Bachelor's Degree plus two years of directly related experience OR an = Associate Degree plus six years of directly related experience OR a valid = credential.
DESIRABLE QUALIFICATIONS: Master's or PhD Degree in a Biological = Science; Experience in teaching / practice of Electron Microscopy
COURSES TO BE TAUGHT:
Introductory Techniques for Transmission Electron Microscopy (EM21) This is a lecture/lab course which includes beginning Transmission = Electron Microscopy dealing with the alignment and operation of the TEM, = vacuum techniques, photographic techniques, as well as the preparation of = particles and replicas for viewing in the TEM. Includes individual = training in the use of the TEM, preparation techniques, and written and = oral reports. (Lec - 2 hrs; Lab - 3 hrs/wk)
Biological Ultrastructure (EM28) Course contents include specific information about the fine structure and = function of cells and tissue at the ultrastructure level. Videos, slides = and micrograph examination will be correlated with the lectures so that = students will learn to recognize the fine structure of cells and tissues = in relationship to their function. (Lec-2 hrs/wk)
Advanced Techniques in Biological Electron Microscopy (EM37) Course contents include lecture and laboratory which covers advanced = techniques for biological specimen preparation in TEM including an = advanced research project.( Lec - 1 hr; Lab - 6 hr/wks)
** Current Microscopies: Optics, Theory and Application (EM30) Course contents include information related to the physical laws and = applications of the various types of current microscopies e.g. = TEM,SEM,FIB, AFM, and confocal microscopy, as well as other current = topics e.g. asbestos analysis, lab design, etc. (Lec - 2 hrs; Lab - 3 = hrs/wk) ** This course is negotiable. A portion of this assignment would be = paid as an additional overload.
The above teaching load would not require development of new courses as = these courses have been fully developed. Course materials are available = to aid in the instruction. TERMS OF EMPLOYMENT: Full semester, Non-tenured track position with Full = Benefits (Medical, Dental, & Vision). SALARY for Fall 98 semester: BA/BS w/ 2 yrs experience or AA w/ 6 yrs experience$17,498 - 25,333 for Fall 98 sem
MA/MS$19, 219 - 28,137 for Fall 98 semester
PhD$ 22,528 - 31,739 for Fall 98 semester
Stockton, CA is 90 miles Northeast of San Francisco and 50 miles South of = Sacramento. It is in lush, green, San Joaquin Valley. We do not suffer = from the high rent districts of San Francisco and Silicon Valley, both of = which are our neighbors.
Additional SJDC Microscopy Technology Program Information available at = http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html/
If you do not contact me in Atlanta, you can contact Human Resources at = the below number however we are up against an extremely tight deadline = and a search committee that may only be available during the Atlanta = meeting. I will have official applications at the meeting and will fax = them to Human Resources.
APPLICATION: Contact Human Resources at 209/954-5056. Human Resources San Joaquin Delta College Admin. Bldg. Rm 202 5151 Pacific Ave Stockton, CA 95207
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Janet, you can either spend lots of dosh on a tripod polisher and polishing system, or do the following;
1) Get some glycothalate (?spelling) thermoplastic wax, such as 'crystalbond' (lots of electron microscopy suppliers sell it).
2) Get some epoxy - anything will do, 5 minute (Devcon) saves a lot of time
3) Glue up a 'sandwich' of three pieces of silicon using the epoxy, with the die you want in the middle. Do it one at a time, using as little glue as possible. Make sure everything is really clean, and squidge the samples about from side to side to squeejee the excess glue away. You can speed up curing on a hotplate. Make sure that all the pieces line up on one edge so that it stands on edge for when you section it.
4) Mount the sandwich, edge on, in the wax (melts at about 120C, nice and runny at 180C) in the centre of a standard polishing platen. Put three blocks of silicon round the edge.
5) Grind till flat on 1200 grit, polish for a few seconds on 2400 grit, fine polish on 1um diamond and a short nap felt pad (slow rotation) for 10 mins.
This is essentially tripod polishing without spending any money. If you want to hit a specific area, you can use a piece of glass instead of the top silicon piece, or even a small piece of silicon so you can see the uncovered part of the die and work out where you are from a (previously taken!) photo.
Let me know if you want any more details,
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK
I think that it is absolutely worthwhile to get a film graphics recorder. While computers with video projectors are probably the wave of the future, at most presentations that I attend, there is the old Kodak Carousel and the 35 mm slides being projected. While a lot of commercial enterprises have the new computerized slide shows, I still believe that 35 mm slides are going to be around for a long time to come, especially in academic institutions. Not every institution nor department can afford the notebook and video projector just to be state of the art in presentation technology. Nor can many investigators for that matter.
I get called on a lot to either shoot 35 mm slides on my film graphics recorder for presentations and often at short notice.
The other thing that you should do as regards the need in your own facility is to ask your customers (the investigators at UCSC) if they use 35 mm slides in their presentations in order to gauge the need for a film graphics recorder. I will bet that the majority of them do. Their Powerpoint presentations as well as any digital microscopic images (confocal or otherwise) can easily be shot on 35 mm slide film with a film recorder.
Matthew J. Schibler Ph.D. Imaging Facilities Core Coordinator UCLA Brain Research Institute 73-384 CHS 951761 Los Angeles, CA 90095-1761
(310) 825-9783 FAX (310) 206-5855 E-mail: mschibler-at-bri.medsch.ucla.edu http:/btrip.mednet.ucla.edu/bri/homepage.htm } ---------- } From: jmkrupp-at-cats.ucsc.edu[SMTP:jmkrupp-at-cats.ucsc.edu] } Sent: Tuesday, July 07, 1998 3:40 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: LM/EM Film recorders? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Dear List: } } In this age of digital imaging, is a film recorder still useful? The } last } presentation I went to was all PowerPoint 'slides' but shown directly } from } a computer to a video projector, no film to be seen. } } I am trying to decide if we should get one, to transfer digital images } back } to film and/or to make slides for presentations etc. If you have some } opinions or experience with these things, could you let me know. } } Thanks. } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } **Area code changing to } 831 as of 7/11/98** } }
One feature of film that has not been mentioned is its archival qualities. It is quite likely that digital presentations will not be very accessible in five years due to hardware and software changes. Properly processed film should last more than 50 years--probably more than 100 years. A slide can be viewed without any eletricity--hold it up to the sky! Try that with a Zip disk.
Has anyone used Image Space Software for looking at dual channel "single section" images. Specifically using the dual channel overlays with two colours. I may be missing something but I cannot get the dual or stereo applications to work with single images. Probably this is not possible with this software it seems to be looking for depth information.
Also which antifade agent do people recommend for fluorescent antibody probes for use with confocal.
I've been following the thread and I have two cents to add about something that I don't think has been touched upon.
I gave an internal presentation that had TEM images that were acquired from a Gatan 1024 x 1024 size CCD camera. The presentation was put together in powerpoint and printed out on a sub-dye printer on overhead transparencies. My manager asked me why I didn't do the presentation with the computer/projection system like several other people have recently done very elegantly. I told them that they didn't have images in their presentations and could get away with it. He said "what does that have to do with anything?"
So I told him the following things: 1) the best projection systems today have about 1024x780 pixel resolution. 2) my images are 1024 x1024 in size. If I wanted to show all the available details in the image, I would have to set the zoom on the powerpoint window so that only the image was visible and none of the title info or other stuff that I have on the slide would be visible. 3) on the sub-dye printer which has 300 dpi resolution, the same 1024 x1024 image would be 3.4 inches in size and I could put several images on the same slide which is then projected with all the details of all of the images in the slide.
Now I would prefer to use 35 mm slides when the absolute best quality in the images are required. However, in the last several years, I have converted to using overhead slides because of the ease of producing good quality overheads in a very short period of time and because slide projectors are becoming less and less available in some locations, particularly in the government labs. When I was at the Materials Lab at Wright Patterson Air Force Base, it was an ordeal to find a working slide projector when someone was coming through the lab that had slides. It is about the same here.
Ok, can I have the change left over from my two cents?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
In this age of digital imaging, is a film recorder still useful? The last presentation I went to was all PowerPoint 'slides' but shown directly from a computer to a video projector, no film to be seen.
I am trying to decide if we should get one, to transfer digital images back to film and/or to make slides for presentations etc. If you have some opinions or experience with these things, could you let me know.
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu **Area code changing to 831 as of 7/11/98**
It has become my task to convince our University here of the strategic importance in maintaining sophisticated microscopy, ie Confocal Microscopy, Electron Microscopy etc as a service to research and teaching.
Here in New Zealand the bulk of scientific research funding comes from the government via a small number of government funding agencies. The research climate is rapidly changing. It would seem the government is moving towards a situation where only a specific number of 'identifiable and targeted' research outcomes will be contested for research funding. If your research activity does not fall into one of these 'targeted' areas and you can not get funding from some other source then you are essentially out of the 'research game'.
With regard to major scientific equipment purchases, such as electron microscopes, NMR's etc, this means that new equipment initiatives (and presumably replacement of existing equipment) will become 'end user driven' rather than, as in the past, 'provider push'.
Traditionally Universities here have obtained research funding on the understanding that the infrastructure to do that research was already in place within the university and, if major equipment purchases were required, then funding was based on current and anticipated usage from a number of researchers or groups, with funds coming from a number of different funding sources (most of which have gone now) and including a contribution from the university itself based on a teaching component. This model is now almost gone.
In the new model it would seem that equipment will only be purchased if it is integral to the efforts of a particular targeted outcome. The 'funding for teaching' aspect is yet to be resolved however it would seem that of the income received by universities from the government to teach students, a 'research component' will also be contestable. This makes capital equipment, such as electron microscopes, NMR's etc, quite vulnerable unless the University can be convinced that it is in it's strategic interest to maintain the availability of such technology and techniques (funding of university teaching activity is also probably going to change soon with the possible introduction of a 'student voucher' system).
As I said above it has become my task to convince our University of the strategic importance in maintaining sophisticated microscopy, ie Confocal Microscopy, Electron Microscopy etc as a service to research and teaching. If such technology and techniques are considered of strategic importance then there is a better chance that the university will continue to maintain such facilities. Obviously I have a vested interest in the long term survival of our Microscopy Unit but I also believe that microscopy as a technique and technology continues to have vital role to play in many scientific disciplines and industrial processes therefore must continue to be taught to our science students.
1. Has anybody out there in 'microscopy land' had to convince their institution of the strategic importance in maintaining sophisticated microscopy within that institution ?
2. If so, I would appreciate some feed back on what arguments you thought worked and what arguments failed.
3. Did you succeed ?
All feedback would be appreciated.
Allan Mitchell
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
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At 09:24 AM 7/8/98 -0500, Darlene Harvey wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
OK, I'll use propane instead of isopentane. Can someone tell me what purity of propane is required? I see 99%, 99.5% & 99.99 (!) % propane available. Airgas also has what they call "Natural grade (Grade 1.6)", 96%. (I assume there's a reason not to use propane containing mercaptans, oil, and who knows what else, from the cylinder of my propane torch or camping equipment. Right?) Can you recommend any US suppliers for 2 to 20 pound bottles? (2 pounds costs almost the same as 20 pounds. I don't need 100 pounds of propane.)
Thanks! Richard
} } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } Quite right, but now the discussion goes to: Which is the better cryoagent and that was a topic here a few months ago. Propane gas liquefied by cooling is a much, much better cryo-agent than is isopentane. Its easy to store in a lab a small gas cylinder with a blunt needle on a bit of tubing as the outlet. With little gas flow rub the needle over the small metal cup that is cooled by liq N2. Soon you will have a couple of ml of liquid propane. Do this in a fumehood, which is a good idea when using solvents too. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
I have a few extra rooms reserved at the Atlanta Marriott Marquis that I will not be using. I know the city is crowded next week so if anyone is looking for a room, please let me know ASAP as I plan to cancel these ext= ra rooms on Friday morning.
Use camping gas and I doubt you will find Any difference. None of the plunge cooling organic cryogens are as good as high pressure freezing (BUT that option although the best is expensive)
Patrick Echlion Multi-Imaging Centre University of Cambridge
On Wed, 8 Jul 1998, Richard Thrift wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } OK, I'll use propane instead of isopentane. Can someone tell me what } purity of propane is required? I see 99%, 99.5% & 99.99 (!) % propane } available. Airgas also has what they call "Natural grade (Grade 1.6)", } 96%. (I assume there's a reason not to use propane containing } mercaptans, oil, and who knows what else, from the cylinder of my } propane torch or camping equipment. Right?) Can you recommend any } US suppliers for 2 to 20 pound bottles? (2 pounds costs almost the same } as 20 pounds. I don't need 100 pounds of propane.) } } Thanks! } Richard } } } } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } } Quite right, but now the discussion goes to: Which is the better cryoagent } and that was a topic here a few months ago. } Propane gas liquefied by cooling is a much, much better cryo-agent than } is isopentane. Its easy to store in a lab a small gas cylinder with a blunt } needle on a bit of tubing as the outlet. With little gas flow rub the needle } over the small metal cup that is cooled by liq N2. Soon you will have a } couple of ml of liquid propane. Do this in a fumehood, which is a good } idea when using solvents too. } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } Great microscopy catalogue, 500 Links, MSDS, User Notes } **************************** www.proscitech.com.au ***** } }
We run courses "in house", in your own laboratory on your own equipment, on all aspects of electron microscopy. Please take a look at our web site for more information, we are able to tune a course to any requirement.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
For a quick review of the new technologies on exhibit at next week's Microscopy & Microanalysis meeting, see the July issue of American Lab: "Focus on Microscopy: What's New at M&M '98", pp. 41-45. (Hot off the presses!)
Extra copies will be available at the meeting at the Microscopy/Microscopy Education Booth (#510).
Richard, you should not have visited that propane variety store, its too confusing. I have used the propane supplied for camping and home stoves. Very likely this contains minor contaminants, but it worked well. Contaminants would change the freezing point in a minor way but I think it very unlikely that they would penetrate and affect cell structure. However, there is an "interesting" research project for some doubter. Please let me know after such "propane purity" research has been accepted for publication. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes service-at-proscitech.com.au *** www.proscitech.com.au
-----Original Message-----
OK, I'll use propane instead of isopentane. Can someone tell me what purity of propane is required? I see 99%, 99.5% & 99.99 (!) % propane available. Airgas also has what they call "Natural grade (Grade 1.6)", 96%. (I assume there's a reason not to use propane containing mercaptans, oil, and who knows what else, from the cylinder of my propane torch or camping equipment. Right?) Can you recommend any US suppliers for 2 to 20 pound bottles? (2 pounds costs almost the same as 20 pounds. I don't need 100 pounds of propane.)
Thanks! Richard
} } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } Quite right, but now the discussion goes to: Which is the better cryoagent and that was a topic here a few months ago. Propane gas liquefied by cooling is a much, much better cryo-agent than is isopentane. Its easy to store in a lab a small gas cylinder with a blunt needle on a bit of tubing as the outlet. With little gas flow rub the needle over the small metal cup that is cooled by liq N2. Soon you will have a couple of ml of liquid propane. Do this in a fumehood, which is a good idea when using solvents too. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
----------------------------------------------------------------------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
OK, I'll use propane instead of isopentane. Can someone tell me what purity of propane is required? I see 99%, 99.5% & 99.99 (!) % propane available. Airgas also has what they call "Natural grade (Grade 1.6)", 96%. (I assume there's a reason not to use propane containing mercaptans, oil, and who knows what else, from the cylinder of my propane torch or camping equipment. Right?) Can you recommend any US suppliers for 2 to 20 pound bottles? (2 pounds costs almost the same as 20 pounds. I don't need 100 pounds of propane.)
Thanks! Richard
} } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } Quite right, but now the discussion goes to: Which is the better cryoagent and that was a topic here a few months ago. Propane gas liquefied by cooling is a much, much better cryo-agent than is isopentane. Its easy to store in a lab a small gas cylinder with a blunt needle on a bit of tubing as the outlet. With little gas flow rub the needle over the small metal cup that is cooled by liq N2. Soon you will have a couple of ml of liquid propane. Do this in a fumehood, which is a good idea when using solvents too. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
We are using LR White for post-embedding staining with colloidal gold. It is absolutely imperative that we get the best possible fixation for synapses. Synapses have a large lipid component. Has anyone tried tannic acid in combination with LR White? If so, how? p-phenylenediamine is known to protect against lipid loss during alcohol dehydration. Has anyone used it with LR White? If so, how? Anyone have any other ideas? We have at our disposal UV light and Progressive Lowering of Temperature techniques, as well as standard oven polymerization. We would so much appreciate any help we can get. We absolutely have to have good fixation of synapses this summer in order to get our next grant (and keep our jobs!) Our antigens tend to be very difficult to locate with Au in material fixed with osmium and embedded in epoxides, which, of course, would give us our best fixation of synapses.
"Can someone tell me what purity of propane is required? I see 99%, 99.5% & 99.99 (!) % propane available. Airgas also has what they call "Natural grade (Grade 1.6)", 96%. (I assume there's a reason not to use propane containing mercaptans, oil, and who knows what else, from the cylinder of my propane torch or camping equipment. Right?) "
I have just constructed an apparatus for freeze spraying in propane, with the help of instructions from Scott Russell, and have been told barbecue gas propane is just fine....in fact the impurities help to reduce the freezing point (I think that's right!) Good job, because the pure stuff costs a bomb!
-- Amanda Wilson awilson-at-sghms.ac.uk Assistant Manager, Electron Microscope Unit, St George's Hospital Medical School, Tooting, London, UK. http://sghms.ac.uk/em tel:? 0181 725 5220 fax:? 0181 725 3326
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{HTML} {BODY BGCOLOR=3D"#FFFFFF"} Richard wrote:
{P} "Can someone tell me what purity of propane is required?=A0 I see 99%, 99.5% & 99.99 (!) % propane available.=A0 Airgas also has what they c= all "Natural grade (Grade 1.6)", {BR} 96%.=A0=A0 (I assume there's a reason not to use propane containing {BR} mercaptans, oil, and who knows what else, from the cylinder of my {BR} propane torch or camping equipment.=A0 Right?) " {BR} =A0
{P} I have just constructed an apparatus for freeze spraying in propane, with the help of instructions from Scott Russell, and have been told barb= ecue gas propane is just fine....in fact the impurities help to reduce the fre= ezing point (I think that's right!)=A0 Good job, because the pure stuff costs a bomb! {BR} =A0
} OK, I'll use propane instead of isopentane. Can someone tell me what } purity of propane is required?...
In my experience, commercial grade propane, such as is sold for use in your barbecue, propane torch and campstove, works perfectly well for plunge freezeing or propane jet freezing. The major differences that I found between chemically pure propane (99%) and commercial grade propane were of course due to the impurites in the latter. For example: (a) the viscosity of commercial grade propane is higher, so I had to enlarge the bore of the propane jets to obtain the same jet velocity (and thus freezing quality), and (b) the freezing point of commerical grade propane is depressed several degrees, and the higher viscosity may slow nucleation, so that I had more time to prepare my specimen before the propane froze. I have also saved a bundle of money and a lot of trouble (since it's so readly available) with the commercial grade stuff. FYI, I have been using 20 lb containers for jet freezing and campstove containers for plunge freezing.
Bill McManus asked: } We recently purchased a microwave system for tissue fixation/embedding. } Unfortunately, the instruction book that we ordered on it's use did not } come and we now find out that it is no longer available (through Ted } Pella). Does anyone have any information on procotols or know of any good } reference books on microwave fixation and embedding? Any information that } will get us going would be greatly appreciated.
I'm not certain which specific book Bill meant, but we sell the Kok & Boon _Microwave Cookbook for Microscopy_ and Gary Login's _Microwave Toolbook_. We also have several microwave processing and microwave fixation procedures at our web site (address in my .sig, below).
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
Just a quick note to say that my experience with propane freezing has been very similar to that of John Gilkey's.
---------------------
" In my experience, commercial grade propane, such as is sold for use in your barbecue, propane torch and campstove, works perfectly well for plunge freezeing or propane jet freezing. ......................" J. Gilkey
"Breakthroughs in biology are preceded by breakthroughs in microscopy."
I cordially invite you to come see Molecular Imaging's technology breakthrough in microscopy, which may dramatically change and speed up the processes in drug discovery.
This "Environmental / In Vitro Atomic Force Microscopy" (AFM) can deliver nanometer resolution time lapse imaging and physical property characterization.
Please come to scientific talks at The Protein Society, 12th Symposium in San Diego, July 25-29, 1998, on this technology and its revolutionary applications: - Prof. Stuart Lindsay, ASU; Oscillating Probe AFM Study of Titin Unfolding, Mon. July 27, 1998, 10:15AM - Dr. Peter Hinterdorfer, University of Linz; Austria (Molecular Recognition Force Microscopy) Antibody-Antigen Recognition Detected By Ultrasensitive Force Microscopy Sunday July 26, 1998, Molecular Recognition, Poster Session, 3:15 - 5:30PM
Or come for a live demonstration by Molecular Imaging at exhibition booth # 613.
If you can not join us in San Diego, please visit us at http://molec.com/BIO-AFM/protein
Respectfully
George Sibbald
PS: If you can not get to Protein come to the Microscopy Society of America Show in Atlanta, July 13-16, 1998 ____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; "Innovating Probe Microscopy" 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Dear All: We are seeking to purchase a TEM simulation software named Mc Tempas runed on Mcintosh computer. We appreciate it if any one can provid the information about where we can buy it, or suggest a better one that can perform interface structure simulation with supercell generation option. Thank you.
---------------------------------------- Dr. Dai Jiyan IMRE National Univ. of Singapore 10 Kent Ridge Crescent Singapore 119260 ----------------------------------------
I am interested in touring the CDC in Atlanta while attending the M&M98. Are there any official tours set up for this? It would be very interesting if we could have a tour geared towards microscopists. Is any one interested?
Sally Burns
Center for Electron Optics burnssal-at-pilot.msu.edu (517) 355-5004
Attention Poor and Desparate EM facilities: -I have three DENTON 515-DV available, used, operational, crated and ready for surplus to any Govt agency who wants it. Just let me know and get your GBL ready. If there are no Gov't takers, then Universities, then Schools are eligible. -I also have two LN2 tanks, on wheels, probably 25L size. Ready to go. -Misc LKB Ultramicrotome repair parts, like fuses, washers, ETC, freebies. -Eight LAB6 filaments for a JEOL 35, never used. You will need the ultra-vacuum pump for these. We never did buy one. -Small Sorvall Histo-knifemaker. That's it for now. I will ship small items today, or after the EMSA meeting. If you are there, I will bring them for you. BIG items will be shipped when the shipping paper work is completed. Please use E-Mail, so I will have a record of the time and date that you responded. Most equipment has service record and instructions. Thank you, Thomas A Baginski, USUHS, Bethesda, MD 20814 Email: tombg-at-bictom.usuf1.usuhs.mil or Voice phone for specific questions 301 295 5691
HILDEGARD CROWLEY wrote: } } Hi, } } We are using LR White for post-embedding staining with colloidal } gold. It is absolutely imperative that we get the best possible fixation for synapses. Synapses have a large lipid component. Has anyone tried tannic acid in combination with LR White? If so, how? } p-phenylenediamine is known to protect against lipid loss during } alcohol dehydration } snip {
Hildy:
I'm pretty sure p-phenyleaminediamine only protects against lipid loss when used after osmium. Sorry I don't have more to offer.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
I would like to look at the cross section of silicon nitride right next to the fracture surface. There is not enough room for dimpling . I was considering tripod polishing, creating a wedge on the side face perpendicular to the fracture surface and hopefully I will get a thin region that is close enough to the fracture. I would appreciate comments or suggestions from people who have tried this.
Hildy, You did not indicate whether it is possible to perfuse fix your material. If so, you might want to check the reference "Liposits, ZS etal (1986). A combined light and electron microscopic immunocytochemical method for the simultaneous localization of multiple tissue antigens. Histochemistry 85:95-106"
I worked with Zsolt on this publication which involved perfusion of rat brains with high and low pH paraformaldehyde, vibratoming the brains, immunocytochemical reaction using the PAP-DAB method, OsO4 fixation and finally embedding in epoxy. It gave very good localization and excellent ultrastructure since membranes were fixed with osmium after ICC. We worked out a simple method of flat embedding the desired areas of the sections between plastic coverslips and then serial sectioning them so we could trace the synapses.
I'm sure that Zsolt has worked out a method to use colloidal gold since that time....probably using nanogold coupled with silver intensification. You should be able to do a literature search and find more recent publications.
Although I do not presently work with neuronal tissue, I would be happy to dig back into my notes after returning from MSA. Do feel free to contact me if you want additional information.
Debby Sherman =================================================== Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Hi,
We are using LR White for post-embedding staining with colloidal gold. It is absolutely imperative that we get the best possible fixation for synapses. Synapses have a large lipid component. Has anyone tried tannic acid in combination with LR White? If so, how? p-phenylenediamine is known to protect against lipid loss during alcohol dehydration. Has anyone used it with LR White? If so, how? Anyone have any other ideas? We have at our disposal UV light and Progressive Lowering of Temperature techniques, as well as standard oven polymerization. We would so much appreciate any help we can get. We absolutely have to have good fixation of synapses this summer in order to get our next grant (and keep our jobs!) Our antigens tend to be very difficult to locate with Au in material fixed with osmium and embedded in epoxides, which, of course, would give us our best fixation of synapses.
So long, Hildy
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Fellow microscopists:
I am looking for suggestions/references on doing immuno on lens from human eyes. For some reason I am under the impression that the texture of the lens may require special fixation and penetration of Ab's may be difficult. Does it become brittle under certain conditions, i.e. would paraffin or cryo- sectioning be better - that, of course, may depend partly on the Ab's.
Also, does lens contain any endogenous autofluorescence?
Thank you for any assistance.
Judy Trogadis Eye Research Institute and University of Toronto Toronto Hospital, Western Div. 399 Bathurst St. Toronto, Canada M5T 2S8
Hi Folks: I want to make some resin casts for SEM of murine skin. two questions, 1: which is the least viscous resin that works well (I need to get at the capillary microstructure) 2: which is the best perfusion method for this type of perfusion (we normally perfuse intracardially) Of course I would welcome any other tips from the wise on this method Thanks a lot Simon
----------------------------------------------------------------------- Simon C. Watkins Ph.D. M.R.C.Path Associate Professor Director, Center for Biologic Imaging University of Pittsburgh Pittburgh PA 15261 tel:412-648-3051 fax:412-648-8330 URL: http://sbic6.sbic.pitt.edu
Sorry if I am sending this message a second time, but I suspect the original message was not ssent out.
Fellow microscopists:
I am looking for suggestions/references on doing immuno on lens from human eyes. For some reason I am under the impression that the texture of the lens may require special fixation and penetration of Ab's may be difficult. Does it become brittle under certain conditions, i.e. would paraffin or cryo- sectioning be better - that, of course, may depend partly on the Ab's.
Also, does lens contain any endogenous autofluorescence?
Thank you for any assistance.
Judy Trogadis Eye Research Institute and University of Toronto Toronto Hospital, Western Div. 399 Bathurst St. Toronto, Canada M5T 2S8
Listers-I thought most of you would appreciate the moral of this story. I know it happens quite frequently.
Paula :-)
} } } } This is a weird but true story (with a moral) } } } } } } } } A complaint was received by the Pontiac Division of General Motors: } } } } } } } } "This is the second time I have written you, and I don't blame you } } for } } } } not answering me, because I kind of sounded crazy, but it is a fact } } } that } } } } we have a tradition in our family of ice cream for dessert after } } dinner } } } } each night. But the kind of ice cream varies so, every night, after } } } } we've eaten, the whole family votes on which kind of ice cream we } } } should } } } } have and I drive down to the store to get it. } } } } } } } } It's also a fact that I recently purchased a new Pontiac and since } } then } } } } my trips to the store have created a problem. You see, every time I } } } } } buy } } } } vanilla ice cream, when I start back from the store my car won't } } start. } } } } If I get any other kind of ice cream, the car starts just fine. I } } want } } } } you to know I'm serious about this question, no matter how silly it } } } } sounds: } } } } } } } } 'What is there about a Pontiac that makes it not start when I get } } } } vanilla ice cream, and easy to start whenever I get any other } } kind?'" } } } } } } } } The Pontiac President was understandably skeptical about the letter, } } } } } but } } } } sent an engineer to check it out anyway. The latter was surprised } } to } } } be } } } } greeted by a successful, obviously well educated man in a fine } } } } neighborhood. He had arranged to meet the man just after dinner } } time, } } } } so the two hopped into the car and drove to the ice cream store. } } } } } } } } It was vanilla ice cream that night and, sure enough, after they } } came } } } } back to the car, it wouldn't start. } } } } } } } } The engineer returned for three more nights. The first night, the } } man } } } } got chocolate. The car started. The second night, he got } } strawberry. } } } } } } } } The car started. The third night he ordered vanilla. The car } } failed } } } to } } } } start. } } } } } } } } Now the engineer, being a logical man, refused to believe that this } } } } man's car was allergic to vanilla ice cream. He arranged to } } continue } } } } his visits for as long as it took to solve the problem. } } } } } } } } And toward this end he began to take notes: he jotted down all sorts } } } } } of } } } } data, time of day, type of gas used, time to drive back and forth, } } etc. } } } } } } } } In a short time, he had a clue: the man took less time to buy } } vanilla } } } } than any other flavor. Why? The answer was in the layout of the } } } store. } } } } Vanilla, being the most popular flavor, was in a separate case at } } the } } } } front of the store for quick pickup. All the other flavors were kept } } in } } } } the back of the store at a different counter where it took } } considerably } } } } longer to find the flavor and get checked out. } } } } } } } } Now the question for the engineer was why the car wouldn't start } } when } } } it } } } } took less time. Once time became the problem - - not the vanilla } } ice } } } } cream - the engineer quickly came up with the answer: vapor lock. } } } } } } } } It was happening every night, but the extra time taken to get the } } other } } } } flavors allowed the engine to cool down sufficiently to start. } } } } } } } } When the man got vanilla, the engine was still too hot for the vapor } } } } lock to dissipate. } } } } } } } } Moral of the story: even insane looking problems are sometimes real. } } } } } } } } A better moral: chocolate ice cream cures vapor lock! } } } } } } } } } } } } }
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207
Does anyone know of a list server or organization similar to MSA for on-line exchange of information to people in the Molecular Biology field? Thanks in advance for any suggestions.
Patrice Abell-Aleff Electron Microscopy Core Facility Mayo Clinic 200 1st Street SW Rochester, Mn. 55905 phone: 507-284-3148 fax: 507-284-9349 e-mail: abellaleff.patrice-at-mayo.edu
There is an opening for a TEM technician at Seagate in the Minneapolis area. Interested individuals please reply directly to "Deb", at address shown at the bottom of the position description by fax or by US mail. Please do not reply by email. Thank you.
Augusto Morrone Seagate Technology Augusto_A_Morrone-at-notes.seagate.com (612)844-5838
Position Open at Seagate Technology: TEM Technician. A TEM technician position is open at Seagate Technology, RHO, in Bloomington, MN. Applicants should have at least a 2-year technical college degree, or equivalent experience, in TEM basic operation and sample preparation techniques in the physical sciences. The main duties involve TEM sample preparation using standard grinding, polishing, dimpling and ion milling techniques, operation and maintenance of sample preparation and darkroom equipment, and may include operation of the TEM, and handling TEM data in the form of negatives and digital images. The TEM facility is installing a Philips CM200 and is projecting the purchase of a FEG TEM within a year. Seagate offers an attractive benefits package and salary commensurate with experience. Please send your resume to:
Seagate Technology Staffing Department 7801 Computer Ave South Bloomington, MN 55435 Fax: (612) 844-7008 Attn: Deb
I am going to purchase a digital camera for general lab usage. I need a digital camera that has a capacity of producing high-quality outputs from both color and B/W photos (the resolution has to be 1024 x 800). Your kind suggestions and advice on the right make and good deal in the market will be appreciated.
Thank you very much for your help.
H. You ********************************************************************* Hong You, Ph.D Dept. of Cell Biology College of Medicine University of Cincinnati Cincinnati, OH 45267-0521 Voice: (513) 558 3709 Fax: (513) 558 4454 email: youhg-at-email.uc.edu *********************************************************************
Dear colleagues, I am trying to find a relatively fast method to count synapses on rat brain sections. I have done immunohistochemistry, but no matter how thin I cut, the synapses are so numerous that it is impossible to count. In old papers they used to do densitometry, but I don't think that is accurate way of counting. I have tried on fresh- frozen or perfused brains embedded in paraffin. I also tried to plastic embed and attempted to count under LM using oil immersion objective. Also tried to do immuno on LR White embedded thin plastic sections, which didn't work very well. Everything to avoid using the EM (too much time consuming, considering that I have six groups of different animals to compare). I would very much appreciate receiving your suggestions. Thank you, Lilith
G'day all from Hot and Muggy Atlanta (at least from a Chicagoan's perspective).
Just a single announcement that this is the Microscopy &Microanalysis 98 Meeting week. I'm in Atlanta now and hopefully all will run smoothly on the Listserver.
Live video and Video Conferences are being broadcast/held from the Meeting Site, in addition a daily Newsletter of events is being posted all to the MSA WWW site.
You may login and virtually join the conference at the MSA URL of:
Dear all, As you will have noticed, there have been two recent ads posted to the list from non members (one for a sex site, and one for an email Marketing program). Whilst the vendors who subscribe to this group almost always behave in a responsible manner by sticking to the charter and not sending overtly commercial mailings and Nestor deals with the occasional abuse, we have no control about non list members who post onto the list.
I am aware that some other discussion groups use software that blocks postings from non list members. I suggest that it may be necessary for Nestor to configure the software that is used for this list to prevent posting from non list members.
What do other people think.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
On 9 Jul 98 at 14:21, The slowly moving finger of Mandayam V. Parthasarathy wrote:
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- } ---. } } Just a quick note to say that my experience with propane freezing } has been very similar to that of John Gilkey's. } } --------------------- } } " In my experience, commercial grade propane, such as is sold for } use in } your barbecue, propane torch and campstove, works perfectly well for } plunge freezeing or propane jet freezing. ......................" J. } Gilkey } } ******************************************************************* } } M.V. Parthasarathy } Prof. of Plant Biology, Adjunct Prof. of Anatomy (Vet), &
On a completely different note. Has any work been done to characterise the Freon replacement gases ?? I can't find any ready reference to this and would like to find a safe replacement to Freon 12 and 22. Thanks John John Findlay Science Faculty EM Facility. Edinburgh University. Daniel Rutherford Bldg. Kings Buildings. Edinburgh EH9 3JH. tel. 0131-650-5344 fax. 0131-650-6563 John.Findlay-at-ed.ac.uk
Try Muller T, Moser S, Vogt M, Daugherty C & Parasarathy MV (1993). Optimisation and application of jet freezing. Scanning Microscopy 7, 1295-1310. Using HCFC 124 (SUVA 124-CHClFCF3) with thin titanium supports, cooling rates were obtained similar to those from propane and standard copper supports. This was suggestedt (I hesitate to plug this !!) in Ryan KP (1992) Cryofixation of tissues for electron microsocpy: a review of plunge cooling methods. Scanning Microsc. 6, 715-743. See next-to-last page , p. 742 in Discussion with Rerviewers section..
} I suggest that it may be necessary for } Nestor to configure the software that is used for this list to prevent } posting from non list members. } } What do other people think. Ian:
Well put, Ian. I agree. The sex post was the pits.
Blystone in Texas
Robert V. Blystone, Ph.D. {RBLYSTON-at-Trinity.edu} Professor of Biology Trinity University San Antonio, Texas 78212 210.736-7243 210.736-7229 FAX
Just use the delete key. Blocking postings from non-listers is a real drag. We have had many many many more legit postings from non-list members than from the ocational spammer. And the thing is, the spammers will co-evolve and get by whatever you try and throw at them. A lot of folks don't like to join because of the high volume of messages that are germane in general but not to them in particular. Just use your delete key. My 2 to the tenth electrons, Tobias } Dear all, } As you will have noticed, there have been two recent ads posted to the list } from non members (one for a sex site, and one for an email Marketing } program). Whilst the vendors who subscribe to this group almost always } behave in a responsible manner by sticking to the charter and not sending } overtly commercial mailings and Nestor deals with the occasional abuse, we } have no control about non list members who post onto the list. } } I am aware that some other discussion groups use software that blocks } postings from non list members. I suggest that it may be necessary for } Nestor to configure the software that is used for this list to prevent } posting from non list members. } } What do other people think. } } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Ian MacLaren, Tel: (44) (0) 121 414 3447 } IRC in Materials for FAX: (44) (0) 121 414 3441 } High Performance Applications, email: I.MacLaren-at-bham.ac.uk } The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ } Birmingham B15 2TT, } England. } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
I totally agree with you. I belong to several other list serv's none of which can be posted to unless you are a registered member. As an aside ... the sex site that was spammed to the list also got to several universities.......
} } As you will have noticed, there have been two recent ads posted to the list from non members (one for a sex site, and one for an email Marketing program). Whilst the vendors who subscribe to this group almost always behave in a responsible manner by sticking to the charter and not sending overtly commercial mailings and Nestor deals with the occasional abuse, we have no control about non list members who post onto the list.
I am aware that some other discussion groups use software that blocks postings from non list members. I suggest that it may be necessary for Nestor to configure the software that is used for this list to prevent posting from non list members.
What do other people think.
Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. { {
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
I concur with Tobias. As obnoxious as they are, the best thing to do with junk mail is ignore it and throw it out. Remember the golden rule for junk mailers - never respond! Even telling them to leave you alone simply validates your mail address on their list which WILL be sold to someone else.
I think the value of this mail list far outweighs the junk mails I get because of it!
I hate when we get bogged-down in all of these "administrative" discussions, but since this could affect the list, here's my opinion:
Spamming is the e-mail version of junk-mail, and I haven't heard of any completely proven manner of removing junk mail without risking the reliability of the "real" mail. When you go through your mail at home, I'm assuming that you don't open everything mailed to you. You can guess that if it promises a million dollars, or has the mark of some credit-card company that you have never dealt with, you know its an ad, and can throw it away immediately, and not lose sleep over it.
My philosophy is: If the subject line doesn't have a topic related to my research, THEN I IMMEDIATELY DELETE IT, even if someone forgets to put a subject line. The subject line is your "filter" that only requires a couple of your own brain cells to activate. Let's face it: I don't know any e-mail software that doesn't show the subject line before opening messages, and if you're worried about spamming, I don't think I've seen any "spam" that didn't have a suspicious sender address (No professional that I know of would send and e-mail from "Lisa" with no last name!), or a suspicious or missing subject line.
This listserver is a forum for discussion, so I have responded to this topic, but let's please not get away from the SCIENTIFIC DISCUSSIONS!
Thank you for listening.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan USA Note: This message is my opinion and has nothing to do with my employer. } -----Original Message----- } From: Ian MacLaren [SMTP:I.MacLaren-at-BHAM.AC.UK] } Sent: Monday, July 13, 1998 5:57 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ads from non list members } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Dear all, } As you will have noticed, there have been two recent ads posted to the } list } from non members (one for a sex site, and one for an email Marketing } program). Whilst the vendors who subscribe to this group almost } always } behave in a responsible manner by sticking to the charter and not } sending } overtly commercial mailings and Nestor deals with the occasional } abuse, we } have no control about non list members who post onto the list. } } I am aware that some other discussion groups use software that blocks } postings from non list members. I suggest that it may be necessary } for } Nestor to configure the software that is used for this list to prevent } posting from non list members. } } What do other people think. } } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Ian MacLaren, Tel: (44) (0) 121 414 3447 } IRC in Materials for FAX: (44) (0) 121 414 3441 } High Performance Applications, email: I.MacLaren-at-bham.ac.uk } The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ } Birmingham B15 2TT, } England. } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ }
Another method that does require some practice but is faster than the jigs and glass wheels is a very simple 2 step method that can be done when time is crucial. Obtain:
1) Pencil size XACTO knife holder. 2) 600 grit SiC paper. 3) MICROCLOTH - polishing cloth from Buehler Ltd. 4) 0.05 micron deagglomerated alumina or 0.02 micron colloidal silica.
The procedure is as follows:
1) Insert the die of interest into the end of the XACTO. 2) With a polish wheel rotating ( in the CCW direction with 600 grit paper, hold the die and holder assembly at 45 degree angle and begin to slowly grind (on the right hand side of the wheel) to the area you desire to cross section. You must frequently monitor the progress with an optical microscope. 3) Once you are 10 - 15 microns away (or so) from the area of interest, stop grinding. Rinse well and dry. 4) Mix up a slurry of 0.05 micron powder and apply to a MICROCLOTH. 5) Begin polishing the die on the left hand side of the wheel at a 90 degree angle to the plane of the wheel. Again, frequent inspection of the progress is needed. The colloidal silica can be used if a very fine artifact free finish is needed, but note that the silica will NOT polish tungsten plugs. When using the colloidal silica, try to find a polish pad that is made of spongy material.....i.e. neoprene or some such, the silica works with MICROCLOTH but a sponge-like material works better. The CMP supplier RODEL has some nice 8" CMP pads that fit the bill. 6) Rinse, clean and dry the specimen and then delineate layers with the appropriate acids
Hope this helps out. We use both methods in our lab but when time is crucial and the volume is large this technique saves the day.
John Staman Consulting FA Engineer Analytical Services Lab Symbios Inc. Colorado Springs, CO. 719-573-3282 ---------- -----------------------------------------------------------------------.
We are currently trying to establish capability in our lab to look at die cross-sections. In previous work we mounted them in epoxy or acrylic and coated the samples, but I'd like to be able to do this without mounting and coating. I've been told that there are simple polishing fixtures one can by to do this, but I can't seem to find where to buy them. Perhaps someone out there could suggest a source? Thanks,
Janet Rice MCC Senior Member Technical Staaff rice-at-mcc.com 512-338-3266
I agree with Tobias that at the moment, there are not sufficient violations of the rules to prevent legitimate postings from non-members. However, this could change. The net is still in its infancy. Ciao for now, Ken
I strongly disaggree with those of you who would simply roll-over and live with junk e-mail posted to a private e-mail list. Will you still be hitting the delete key when half the postings were never invited? ... or will you be unsubscribing?? I belong to several lists and monitor one myself and can attest to this list being especially vunerable (... for some reason ...). This list's monitor really should look into different list server software.
[to Nestor: I really do understand you have a real job ... and do sympathize with some of this not being under your control ... that is, I don't get to choose the list software this university uses ... but please pass these concerns on to your lists-meister ...]
... my $0.02 :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I agree that these Spam postings are an annoyance. It would be nice if one could reply to them and overload their server. I know in fact that this is a bad idea, as it gobbles bandwidth, but it would just feel good to give them a dose of their own clutter. I know there is a web site to deal with this issue, I will find it and if there is any useful information I will post to the server. For now the delete key sounds best and fastest, if not the most emotionally satisfying.
-- Respectfully, Bob ( Robert G. ) Lawrence Failure Analyst Motorola Phoenix Corporate Research Lab 2100 E. Elliot Rd. MD EL-703 Tempe, AZ 85284-1806 Phone: 602-413-5848 Fax: 602-413-4952 Pager: 1-800-759-7243 PIN 834-2458
} [snip] } } ...I suggest that it may be necessary for } Nestor to configure the software that is used for this list to prevent } posting from non list members. } } What do other people think. }
It is easy to set most software to not allow non-members to post to a list. However, turning off automatic subscription and then vetting would-be subscribers is a tremendous amount of work. Since there is no vetting for subscription to this list, there is no protection from folk automatically subscribing to post an ad. I'll even bet that's how those ads got on here.
The degree of security one imposes on a list is directly related to the amount of work necessary to administer and maintain the list.
While I subscribe to a couple of lists where subscription is not automatic, they all cater to rather small groups. It's a bit much to ask Nestor to perform the vetting function. Until such time as Nestor gets extra pay to maintain the list and until such time as we decide to pay for his infrastructure, I suggest that Nestor can do whatever the hell he wants.
Dear all, As you will have noticed, there have been two recent ads posted to the list from non members (one for a sex site, and one for an email Marketing program). Whilst the vendors who subscribe to this group almost always behave in a responsible manner by sticking to the charter and not sending overtly commercial mailings and Nestor deals with the occasional abuse, we have no control about non list members who post onto the list.
I am aware that some other discussion groups use software that blocks postings from non list members. I suggest that it may be necessary for Nestor to configure the software that is used for this list to prevent posting from non list members.
What do other people think.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Nestor gave a report on this very problem yesterday to MSA Council, sponsors of this List. He is well aware of the problem but the solutions are difficult. Nestor's a little busy right now here at M&M'98 in Atlanta (check out the WWW site), but will probably reply himself when he has a chance.
Ernie Hall MSA Secretary
---------- From: Ian MacLaren[SMTP:I.MacLaren-at-BHAM.AC.UK] Sent: Monday, July 13, 1998 5:57 AM To: Microscopy-at-sparc5.microscopy.com Subject: Ads from non list members
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Dear all, As you will have noticed, there have been two recent ads posted to the list from non members (one for a sex site, and one for an email Marketing program). Whilst the vendors who subscribe to this group almost always behave in a responsible manner by sticking to the charter and not sending overtly commercial mailings and Nestor deals with the occasional abuse, we have no control about non list members who post onto the list.
I am aware that some other discussion groups use software that blocks postings from non list members. I suggest that it may be necessary for Nestor to configure the software that is used for this list to prevent posting from non list members.
What do other people think.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
One camera that might be suitable for your requirements is the svMicro digital camera. This new camera will be available September 1st; it is a c-mount, resolution of 800x1000, 3 shot color camera, live black and white video preview & operates as a Photoshop plugin. On a MAC the svMicro is a SCSI device; on a PC, it is a parallel device. The CMOS chip is sensitive enough to image bright fluorescent subjects. The price is $2,200 as a c-mount and $2,600 with a package that includes lenses & extension tubes. For more information, please feel free to contact me at telephone 516-773-4305.
Matt Irwin ElectroImage, Inc. 277 Northern Blvd Suite 101 Great Neck, NY 11021
I am in the process of creating a web page for irusers-l, an internet listserv for scientists and conservators around the world who use infrared spectroscopy to study historic and artistic works and other cultural property.
The page will include links to commercial suppliers of FT-IR instrumentation and materials.
If lurking company reps would like to e-mail me company names, contact information, and URLs I'll try to include them in the page.
I'm realizing my PhD and I research about an halophyte, Atriplex nummularia Lindl. I work with plant anatomy and I'm looking for a stain that marks the vesicular hairs that are in this specie. Would you help me? Thanks.
i am working on the localization of antigens being involved in the adhesion/invasion process of pathogenic bacteria. Pre-embedding labelled samples are embedded in a resin which is suitable for perfoming post-embedding labelling afterwards like Lowicryls etc. I would like to know if someone has got any experience in using silver-enhancement kits for enlarging the gold-particles of the pre-embedding labelling with subsequent post-embedding labelling of the same ultrathin section.
- does the used resin has any effect on enhancing the gold-particles of the pre-embedding label
- does the silver-enhancement solution penetrates the entire ultrathin section and developes not only those gold-particles being exposed on the surface of the ultrathin section
- does silver-enhancement allows to perform postembedding labelling afterwards.
In the past, I've seen discussions on the list about the effects of vibrations from roadways, "floods" (both water and liquid nitrogen), ambient dust etc. on electron microscopes, but none have addressed lightning.
It may sound funny, but this is serious business.
It wasn't a direct hit, but it certainly "fried" a good deal of the electronics in our SEM. 2 1/2 weeks ago lightning struck the nearby power plant that supplies us with electricity, wreaking havoc all about in our industrial park. All the local production facilities managed to get back running within hours or a day of the disaster. Our SEM is *still* down, despite repeated visits of the SEM manufacturer's technical service people, who have been having trouble finding all the problems. This does not reflect well on the manufacturer...nor on us, for choosing this manufacturer.
But I'm interested in preventing similar problems in the future. Here in Germany, lightning is not all that frequent and at home I have *never* experienced a power outage due to thunderstorm action (20 years). But I think our power plant in the industrial park may be "lightning prone", since this is the second hit it's taken in the past 5-6 years. (The first time, we had no SEM.)
I'd be interested in suggestions about protecting our SEM from such hits to the power line in the future. Surely there are SEMs in more lightning-prone areas (Natal, South Africa? Southeast US?), where such protection must be routine. Can anybody give me some ideas?
Cynthia Bennett Hoechst Diafoil Germany
************** The opinions expressed here are solely my own and not the fault of my employer. **************
I think that very general "what do you people recommend" inquiries are best sent to the inquirer only and that person could then broadcast a summary. Another camera was touted here, so I wish to advise that Pixera has: 1260x940 pixel, comes complete with lens and other accessories, has new, superior software, offers most pixel/$ (even before the recent substantial price-reduction). The Pixera patent also gives better depths-of-field for it's pixel size than any other camera. A lot of technical info is provided in our online. Disclaimer: Yes, ProSciTech supplies Pixera and I would not have made this rather commercial posting had not another supplier preceded. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
From: H.You {youhg-at-email.uc.edu} To: microscopy-at-Sparc5.Microscopy.Com {microscopy-at-Sparc5.Microscopy.Com}
Lightning strikes and the resulting spikes are a problem, especially for us on the lightning-prone 'Highveld' of South Africa. Ordinary lightning protection diode and voltage dependent resistors are not necessarily fast enough to protect your EM. We use a UPS to protect EM's. You do not need a bigger capacity than one that will carry the microscope for a few minutes, so they are not too expensive. You do, however, need one that fully isolates the microscope from the mains supply. Take care to get one that outputs a clean sine waveform, even under full load - some of the UPS systems produce a pretty nasty square wave when working hard.
Jan Coetzee
} } Hi Listers, } } In the past, I've seen discussions on the list about the effects of } vibrations from roadways, "floods" (both water and liquid nitrogen), } ambient dust etc. on electron microscopes, but none have addressed } lightning. } } It may sound funny, but this is serious business. } } It wasn't a direct hit, but it certainly "fried" a good deal of the } electronics in our SEM. 2 1/2 weeks ago lightning struck the nearby } power plant that supplies us with electricity, wreaking havoc all } about in our industrial park. All the local production facilities } managed to get back running within hours or a day of the disaster. } Our SEM is *still* down, despite repeated visits of the SEM } manufacturer's technical service people, who have been having } trouble finding all the problems. This does not reflect well on the } manufacturer...nor on us, for choosing this manufacturer. } } But I'm interested in preventing similar problems in the future. } Here in Germany, lightning is not all that frequent and at home I } have *never* experienced a power outage due to thunderstorm action } (20 years). But I think our power plant in the industrial park may } be "lightning prone", since this is the second hit it's taken in the } past 5-6 years. (The first time, we had no SEM.) } } I'd be interested in suggestions about protecting our SEM from such } hits to the power line in the future. Surely there are SEMs in more } lightning-prone areas (Natal, South Africa? Southeast US?), where } such protection must be routine. Can anybody give me some ideas? } } Cynthia Bennett } Hoechst Diafoil } Germany }
Prof Jan Coetzee Head: Lab for Microscopy and Microanalysis Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-362-5150 Pretoria 0002 Internet:janc-at-ccnet.up.ac.za South Africa http://www.up.ac.za/science/electron/emunit1.htm
Cynthia Bennett wrote ================================== In the past, I've seen discussions on the list about the effects of vibrations from roadways, "floods" (both water and liquid nitrogen), ambient dust etc. on electron microscopes, but none have addressed lightning.
It may sound funny, but this is serious business.
It wasn't a direct hit, but it certainly "fried" a good deal of the electronics in our SEM. 2 1/2 weeks ago lightning struck the nearby power plant that supplies us with electricity, wreaking havoc all about in our industrial park. All the local production facilities managed to get back running within hours or a day of the disaster. Our SEM is *still* down, despite repeated visits of the SEM manufacturer's technical service people, who have been having trouble finding all the problems. This does not reflect well on the manufacturer...nor on us, for choosing this manufacturer.
But I'm interested in preventing similar problems in the future. Here in Germany, lightning is not all that frequent and at home I have *never* experienced a power outage due to thunderstorm action (20 years). But I think our power plant in the industrial park may be "lightning prone", since this is the second hit it's taken in the past 5-6 years. (The first time, we had no SEM.)
I'd be interested in suggestions about protecting our SEM from such hits to the power line in the future. Surely there are SEMs in more lightning-prone areas (Natal, South Africa? Southeast US?), where such protection must be routine. Can anybody give me some ideas? ===============================================
I think the reason is in temporary increase of mains voltage. Methods of protection: - autonomous generator - most reliable method, - motor-generator with additional protection of the motor, - high-power BACK UPS (with constant converting of mains voltage) with additional protection of input circuits - most modern method. Regards.
Victor Sidorenko, ANTRON Co. Ltd., Moscow, Russia. antron-at-space.ru
For what it's worth, I agree with Gregg. I'm not brain dead yet (despite the long hours) and I can figure out which e-mail I want to read and which I want to delete. "For your eyes only", as a subject line, was a dead giveaway!!!
I'm sure that Nestor has enough to do without having to protect us all from a few pranksters.
} Hello all, } } Does anyone know of a list server or organization similar to MSA for } on-line exchange of information to people in the Molecular Biology field? } Thanks in advance for any suggestions. } } Patrice Abell-Aleff } Electron Microscopy Core Facility } Mayo Clinic } 200 1st Street SW } Rochester, Mn. 55905 } phone: 507-284-3148 } fax: 507-284-9349 } e-mail: abellaleff.patrice-at-mayo.edu
Nestor needn't go to the trouble to monitor subscriptions to the list or block mail from certain domains, but as the 'list owner' he should probably do his part to eliminate the spam at its source (see below), if he isn't already. Unsolicited bulk commerical email is a huge drain on the network bandwidth, disk space and other resources of network providers and SMTP relay sites. Furthermore, unlike bulk postal mail, for whch the sender pays at least part of the cost, the cost of bulk email is paid by us consumers in increased ISP charges, slow netowrks, etc. I started to receive several spams a day in my personal email about two years ago. Aside from the tangible and intangible costs, I consider spamming to be in intrusive and irresponsible use of network bandwidth, so I started sending messages to the postmaster and administrative contact (obtained from a WHOIS lookup, http://rs.internic.net/cgi-bin/whois) of the originating sites (watch for forged entries, though), their ISP, if any, and the postmaster of all relay sites listed in the message header (who can put pressure on the originating site to take action - again, watch for forged entries). By last spring, I had received notification of the termination of the accounts of several dozen of these spammers, and the rate of spamming had dropped to a trickle (1-2 per week). Some might think that by sending these messages, I was only adding to the noise, but while a single bulk mailing consists of thousands of messages, only a handful of people complain, and this is sufficient to alert postmasters to the problem and eliminate it at the source. I have in fact found that postmasters appreciate someone informing them of what is generally unwanted network traffic (for the relay sites) or a violation of the conditions of use (of a provider's services), so that they can deal with it effectively, since it is such a huge drain on their resources. Those spammers who have persisted and been brought to trial have generally been convicted on "theft of services" charges for this innapropriate use of other's equipment and abuse of priveleges. There is some remedial legislation in Congress, but unfortunately it does not go so far as to place the same sanctions on unsolicited commercial email as were placed on unsolicited commercial faxes ($500/message or cost to recipient, whichever is greater); see:
We are selling a JEOL 9000 freeze fracture unit. It needs a little work, but we never use it so we never had it repaired. We will accept the best offer we receive. You will have to pay for the shipping. If you're interested in this lovely putty & orange colored machine, please contact me. I promise that we will clean all of our junk off of it [we've been using it as a workbench ;)].
Looking forward to seeing the floor underneath it,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207
In Tampa Bay, (FL, USA), touted as the lightening capital of the world, = lightening protection is a must. The local TV stations enjoy giving out = frivolous statistics such as the number of surface-to-cloud strikes per = hour. For example, night before last, there were more than 9000 strikes = in a one hour period. Hence, the potential of lightening damage is taken = seriously and has spawned an entire industry of consultants, manufacturers = and installers. The first line of defense seems to be the protection of = the building (every corner, projection etc. has a 12-18" lightening rod = and these are all connected), and there is massive protection on the main = power lines and communication lines. In fact, our computer network lines = are fiber even between floors of the main Hospital here. Most of this = falls under the responsibility of our main Hospital engineer and costs = many tens of thousands of dollars to install and maintain.
Although this is adequate for many applications, sensitive equipment = requires much more and that is even more expensive. I teamed up with the = rest of the Laboratory, and had installed a very large battery powered, = online UPS (takes up a whole room). From this we can run all essential = equipment for about ten minutes in the event of an outage, time enough for = the generators to kick-in. This device also smooths out generator noise, = and brown outs, (low voltage). In extremely simplistic terms, the = incoming current is disassembled on the incoming side, and reconstituted = on the outgoing side. The batteries not only act as a reserve and buffer, = but also a filter. All grounds are on this side of the UPS to prevent = ground loops, et cetera. At the time (about five years ago), this was the = most cost effective way to cover a multitude of bases for a large clinical = laboratory. =20
Intermediate-sized equipment, not in proximity to the large UPS, is on = individual battery powered online UPS. There is a wide range of sizes = available from vendors (for example, APC) for a wide range of dollars. = This is what I would expect is best for a single instrument such as a SEM. = Pay attention to wave shape and ratio for computerized equipment (vendors = can advise). If you cannot locate any local outlets, your network gurus = may be able to point you to their sources. Most network servers have such = equipment installed, albeit smaller than you probably require. The same = vendors usually sell the larger models as well (5 to 35 KW).
Our desktop computers are NOT on these devices. Instead, they are on = individual surge protectors only (for example, Tripplite surge protectors) = with no battery backups. One aspect about computers is that the majority = of strikes come across phone lines so a surge protector with phone line = protection is a must! That is, unless your institution has a switchboard, = which is already protected. At home, I experience at least one or two = such strikes a year that fries the modem protection on my computers. =20
A note of caution, do not place the inexpensive types of surge protectors = in serial, as it is possible to create destructive harmonics between the = devices.
Of course, like insurance, you must gauge the investment and risk with the = cost of protection.
Regards,
Peter O. Steele, Ph.D., PMIAC Dir., Special Anatomic Pathology Unit Pathology & Laboratory Medicine All Children's Hospital Saint Petersburg, Florida 33731-8920 v-mail: 813/892-4465 e-mail: steelep-at-allkids.org
We are thinking of putting uninteruptable power supplies (UPS) on our electron microscopes (Philips & Zeiss TEM's).
Any suggestions or comments?
Thanks - Dennis
------------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: winkler-at-calvin.niams.nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
I am trying to use a quantitative analysis program to analyze EDS spectra collected with TEM. I have 2 questions: (1) I have oxide inclusions in a diamond matrix . I think there are absorption problems (Carbon absorbing oxygen). Is there anyway of getting around it? I am using the "thin-section analysis" as an action. Should I use " bulk sample analysis"?
(2) I am using Nickel grid. So, the spectrum has Ni as an element in it. How do I calculate the formula of the mineral? Do I perform the quantitative analysis without the Ni? Or do I normalize the other elements without taking into account that the Ni is present? Is there anyway of determining if I do have Ni in my sample (or finding out proportions of Ni coming from the grid and from the sample?)
} We are thinking of putting uninteruptable power supplies (UPS) on our
} electron microscopes (Philips & Zeiss TEM's).
}
} Any suggestions or comments?
}
Dennis,
We have the Ferrups unit from Best Power on several instruments. There are two things that I would keep in mind:
1. make sure that you have the capability to handle the power surges that come from motors switching on and off (ie, pumps and compressors). For Best Power, this is the DVR option.
2. make sure that your physical plant people RTFM when installing the unit. For example, Best requires grounding cable that is *at least* the same gauge as the hot and common wiring. This requirement is more stringent than the typical electrical code.
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 7 - October 15, 1998
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2050 (Includes room and board)
Application Deadline: August 4, 1998
Admission application and information: Carol Harnel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratio-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual research problems with the faculty.
} We are thinking of putting uninteruptable power supplies (UPS) on our } electron microscopes (Philips & Zeiss TEM's). } } Any suggestions or comments?
Here's how it works in our lab:
The SEM and TEM are both running on a 220 V AC mains separated from the "ordinary". The separation is done by a 220 V AC generator driven by a 220 V AC electric motor. It seems to be a dull solution but this way we completely eliminate all electric surge and unwanted noise from the mains and besides the inertia of the two rotating iron cores is enough to overcome about 1/10 sec dropouts of the "ordinary" mains supply. We really do not need a UPS (taking into consideration the high power of TEMs and SEMs the UPS should be dimensioned quite big and expensive). During the past more than twenty years we had about two or three dropuouts long enough to shut down the microscopes. And the generator/motor system runs without problems, almost no maintenance, and cheap, even if you take into account the efficiency loss of the system. We even operate some sensitive computer servers and other instruments on this same local and very "clean" mains.
Kris
PS: The above solution also provides extremely effective protection against lightnings, see posting of Peter Steele ---------------------------------------------------------------------------- --------- Kristof Kovacs, Ph.D. Head, Central Laboratory University of Veszprem, P.O.Box 158, Veszprem H-8201 Hungary Phone: +36-(88)-421-684 Fax: +36-(88)-328-643
Hello, Do any of you know where I might obtain a copy of the book "Cell and Tissue Ultastructure- A Functional Perspective" by Patricia C. Cross and K. Lynne Mercer. W.H. Freeman and Co. 1993? It is out of print and I have checked the obvious resources. This is an excellent resource which I would very much like to add to my library.
I just joined the list. I love the infinite complex world. I really want a microscope and am interested in building one if it's cheaper. I've been researching home made telescopes too but cant find any information on making microscopes. Could someone help me? A powerful microscope would be great, like 1000x or larger, if possible (smaller's ok, I'd like at least 100x), but I'm assuming its similar to telescopes in that you can make pretty much any size given your time, money, and experience level. Perhaps someone knows of urls or books relevant?
thanks,
danny
ps- has anyone heard of Ernst Haeckel? He was a biologist who drew images of small complex life forms, here are a few examples of his art... I would love to be able to see these under a microscope
{A HREF="http://www.comptime.com/hoti/Haeckelview.html"} Haeckelview.html at www.comptime.com {/A}
In a message dated 7/15/98 12:28:50 AM US Eastern Standard Time, CALYK-at-aol.com writes:
} Microscopy-at-sparc5.microscopy.com
Hi there, when I was a younger person I used a book called wonders through the microscope to build a projection microscope. Gee, they even had plans in there to build your own arc lamp to use with it........a wonder we didn't burn the house down....
Anyway Check your local library there still may be a copy of this old tome there.
If all else fails maybe we even have an extra copy of it in the library here that we could trade off. The authors were the editors of popular science monthly. Edward Sharpe, Archivist SMECC
An NIH Director's Wednesday Afternoon Lecture Series Event Sponsor: NIH Inter-Institute Mitochondria Interest Group (MIG)
A Day-long Minisymposium=20 Mitochondria: Genetics, Health, and Disease 2 December 1998 Featuring The Wednesday Afternoon Lecture by Dr. Eric A. Schon (Columbia) Molecular Genetics of Human Mitochondrial Disease
Jack Masur Auditorium, Clinical Center, NIH For special accommodation needs call 301-594-5595 CME credit awarded=20 MITOCHONDRIA: GENETICS, HEALTH, AND DISEASE MINISYMPOSIUM 2 DECEMBER 1998
Lectures Venue: Masur Auditorium, Clinical Center, NIH Poster/Exhibitor Venue: Visitor Information Center, Clinical Center, NIH
* 0745 Registration, Poster Set-up, Continental Breakfast in Exhibit = Area * 0830 Dr. David A. Clayton (HHMI): Mitochondrial DNA Control Features * 0905 Dr. William C. Copeland (NIEHS): Avoidance of Mitochondrial DNA = Mutations by DNA Polymerase Gamma * 0940 Dr. Vilhelm A. Bohr (NIA): Oxidative DNA Damage Repair in = Mammalian Mitochondria * 1015 Poster Session/Coffee Break in Product Exhibit Area * 1045 Dr. Tracey Rouault (NICHD): Abnormalities of Mitochondrial Iron = Metabolism and Human Disease * 1120 Dr. Steven J. Zullo (NIMH): In situ Localization of the common = Human 4977bp Mitochondrial DNA Deletion Mutation * 1200 Lunch Break * 1330 Dr. Mariana Gerschenson (NCI): Mitochondrial Genotoxic and = Functional Consequences of Chemotherapeutic Drugs * 1400 Poster Session/Coffee Break in Product Exhibit Area * 1500 Wednesday Afternoon Lecture: Dr. Eric A. Schon = (Columbia):Molecular Genetics of Human Mitochondrial Disease * 1600 Reception/Poster Session in Product Exhibit Area * 1700 Poster Session/Product Exhibition Closes
Continental Breakfast, Coffee Breaks, Lunch Break, and Reception = sponsored by the Technical Sales Association Attendance/Poster Registration Form Mitochondria: Genetics, Health, and Disease Wednesday Afternoon Lecture Series Minisymposium 2 December 1998 Masur Auditorium and Visitor Information Center Clinical Center (Building 10), NIH Bethesda, MD
Note: There is no registration fee to attend this minisymposium! Name: Title: Affiliation: Address: State: Country: Postal Code: Telephone: Fax: E-mail: Web Page URL: I will attend and present a poster I will attend but not = present a poster________ Poster Title: Abstract (Abstract Booklet available at the meeting):
Do you need special accommodations while at NIH?______________ For a map of NIH and area, please check the following Web Site: = http://www.nih.gov/welcome/maps.html WARNING: Parking is limited on = campus, plan to use METRO! A block of rooms at a special meeting rate has been reserved at The = Bethesda Ramada. Call 800-272-6232, or 301-654-2703, before 9 November = 1998. Mention NIH Minisymposium, group # 6210. For other = accommodations in the area, please check the following Web Site: = http://www.patsys.com/ftd/city.cgi?pcityid=3D2521&pcity=3DBethesda Submit advanced registration via Minisymposium web site at = http://www-lecb.ncifcrf.gov/~zullo/migDB/symposium.html or via e-mail to: zullo-at-helix.nih.gov deadline by e-mail is 8 November = 1998=20 or via regular mail to: Steven J. Zullo, PhD postmark deadline is 2 = November 1998 Building 10, Room 2D54 NIH Bethesda, MD 20892
Steven J. Zullo, PhD Laboratory of Biochemical Genetics NIMH-NIH; Bldg. 10, Rm. 2D56; 9000 Rockville Pike Bethesda, MD 20892 301-435-3576; FAX 301-480-9862 zullo-at-helix.nih.gov Mitochondria Interest Group Web Page: = http://www-lecb.ncifcrf.gov/~zullo/migDB/
I was wondering if anyone has experience with the new Leica CPC unit. We are using this for freezing thin films to get virtified ice on holey carbon grids. We recentlly purchased one of these units and I would be extremely grateful for any tips and advice from someone with experience of using the unit for this application. How do you avoid ice contamination?
Many thanks in advance,
Dr Timothy F. Booth Canadian Food Inspection Agency National Centre For Foreign Animal Disease Suite T2300 1015 Arlington St. Winnipeg Manitoba R3E 3M4 CANADA http://www.hc-sc.gc.ca/main/lcdc/web/bmb/fedlab_e.html#toc email tbooth-at-em.agr.ca Tel 204 789 2022 Fax 204 789 2038
There are a number of sources of plans for home made microscopes; however, I really don't think you will save any money over the very low cost microscopes currently imported from China.
Here is the design I consider the most realistic for construction using only hand tools: Curry, Alan; Grayson, Robin F.; and Hosey, Geoffrey R. "Under the Microscope"; Van Nostrand Reinhold, New York, 1982; pp 31-40. (This design is/was featured on the Boston Museum of Science Web page)
I have a few ideas on how to simplify this even further, contact me if you are interested.
What is the current, generally accepted way of disposing of osmium tetroxide?
I use 1% osmium tetroxide in sodium cacodylate buffer. After removing this solution from my tissue, I pour it into a hazardous waste container that is 2/3 filled with corn oil to bind the osmium solution. We've done this for years, but our new hazardous waste/disposal monitor needs to have an official source with exact quantities of oil to osmium tetroxide ratios.
Can you help me? If there is safer, more acceptable method of disposal, your suggestions would be greatly appreciated.
Thank you in advance for you help and guidance.
Lucy Stribling Air Force Research Lab Brooks AFB Texas
I am considering the purchase of an evaporative carbon coater for use on coal clinker specimens that have a history of bad charging under the SEM. The evaporative carbon coaters (carbon rod source) come in two types: high vacuum (10^-5 torr) and low vacuum (10^-2 torr). The high vacuum coaters (with a turbo pump) are significantly more expensive. Are they worth the extra cost?
I'm told that www.abebooks.com is an excellent web site for out-of-print and hard-to-find books in the sciences and other areas. Haven't yet tried it personally, but a friend has had good success.
Good luck.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
We maintain a JEOL 2010 FEG-TEM for which a UPS is regarded by the manufacturer as essential. We use a Phase One Series 700 12.5 KVA unit that was suggested to us by JEOL. It works great and has plenty of capacity (right now it is running the scope and some accessories and only outputing 50% of its full capacity). It and the microscope have been in use for 5 months and with all the power outages due to thunderstorms here in Houston I don't know how we could live without it. If I could I would get one for our other microscope and every other piece of major equipment in our facility. So my thoughts these days are: UPS, Gotta' have it!
Disclaimer: I have no financial interest in any of the abovementioned companies
TTFN
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Roy Christoffersen Materials Science Research and Engineering Center University of Houston 3201 Cullen Houston, TX 77204-5932 roy-at-bayou.uh.edu (713) 743-8273 FAX: (713) 743-2787
I'm told that www.abebooks.com is an excellent web site for out-of-print and hard-to-find books in the sciences and other areas. Haven't yet tried it personally, but a friend has had good success.
Good luck.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
I'm told that www.abebooks.com is an excellent web site for out-of-print and hard-to-find books in the sciences and other areas. Haven't yet tried it personally, but a friend has had good success.
Good luck.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Dear Barna, The science of EDS analysis on a TEM is a less well-characterized than on an SEM. I think that the results are semi-quantitative at best and it is difficult to know the exact conditions of film thickness and other sample characteristics for good analysis. Good quantitative analysis will require some known standards with a composition close to your unknowns. However, in answer to your questions: (1) Carbon will not appreciably absorb oxygen in your oxides. X-rays are absorbed by elements heavier than the emitting element. The absorbtion correction for oxygen in a carbon matrix, particularly in a thin film, will be very small. Use the "thin-film" analysis mode. (2) When you calculate the formula of your mineral, just leave the element of your grid out of the element list. The only way I know to tell if you have Ni in your sample is to use another grid. Copper, nylon, berylium, molybdenum, etc. are all available. Or, if your diamond matrix is strong enough, use a large-mesh grid, slot or single-hole grid so you can do measurements a long way from the grid material and see if your Ni response drops down to a minimal signal. You wrote: } } } Hi everyone, } } I am trying to use a quantitative analysis program to analyze EDS spectra } collected with TEM. } I have 2 questions: } (1) I have oxide inclusions in a diamond matrix . I think there are } absorption problems (Carbon absorbing oxygen). Is there anyway of getting } around it? I am using the "thin-section analysis" as an action. Should I } use " bulk sample analysis"? } } (2) I am using Nickel grid. So, the spectrum has Ni as an element in it. } How do I calculate the formula of the mineral? Do I perform the } quantitative analysis without the Ni? Or do I normalize the other elements } without taking into account that the Ni is present? } Is there anyway of determining if I do have Ni in my sample (or finding } out proportions of Ni coming from the grid and from the sample?) } } Thanks very much for your time and help, } } Please reply to: barna-at-geo.princeton.edu } } } } Barna } Best of luck, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
... i am not quite sure, but I think that there has been a discussion about different dye-sub printers on this list-server... sorry for coming back to the same point, but here is my problem: we are planning to buy a high-end "photorealistic" printer. For certain reasons we tend to a Codonics ... Could anybody explain to me, what the advantage of the additionally implemented "direct thermal" technology (-} Codonics NP1660) is? The main task for our printer would B&W prints. The maximum percentage of colour prints is estimated to be in the region of 10 to 20%; pages/year appr.: 750..1000 ). It would be also very interesting to to have a comparison about the costs per page (Codonics NP1600 versus NP1660) including paper and ribbons, but excluding the investing costs for the printer itself.
Thank You very much in advance! Dipl.Ing.(FH) Gunnar Glasser, Elektronenmikroskopie
The coater type of choice for your application is the LOW vacuum type. For
some applications, low vacuum coater films are not as "good" as high vacuum types. In your case, however, the lower vacuum means more scattering. With increased carbon scatter, the coating is less "line-of-sight" and will better coat rough, convoluted surfaces.
I would also strongly suggest you investigate low vacuum coaters which use a
carbon yarn filament rather than the carbon rod type.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am considering the purchase of an evaporative carbon coater for use on coal clinker specimens that have a history of bad charging under the SEM. The evaporative carbon coaters (carbon rod source) come in two types: high vacuum (10^-5 torr) and low vacuum (10^-2 torr). The high vacuum coaters (with a turbo pump) are significantly more expensive. Are they worth the extra cost?
by imo24.mx.aol.com (IMOv14_b1.1) id NOCPa22587; Wed, 15 Jul 1998 13:45:13 -0400 (EDT) Message-ID: {7b79c6b5.35aceaab-at-aol.com}
If someone knows a contact at popular science mag. perhaps we can get permission to have a file of the projection microscope that plans that are downloadable on the MSA site?! We have the orig. book here that we could scan but hate to get our hands slapped by a publisher! I will leave this idea in the hands of some able diplomat that would like to contact popular science or who ever owns them now!
Sorry for the several postings about the books---"operator error" is to blame. A newly-learned fact: when a message is returned because of an improper primary address, the "cc:" address still goes out just fine. oops....
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Recently there was a message sent out about the Woods Hole microscopy course for the fall. In error I deleted it and would like to have a copy of it. Anyone have it to forward to me?
Hi, can anyone there help me to put the scale bar onto a confocal image? I saved confocal image with scale bar (with overlay option), but once I open the image with any other imaging program, such as PhotoShop, the scale bar is not on the image any more. I wonder what I did wrong and how can I save the confocal file with scale bar as well as make the scale bar occurs when other imaging softwares other than LSM operating software are used. Your help will be greatly appreciated. Thank you.
I have just acquired two ADEM1 SEMs and am now in a quandary as to what to do with them. It is not economical to get them running and sell them as the upkeep costs for a contract is about $30k per year. I could donate them to a school but I have already donated two Hitachi SEMs in the last two years.
They are worth more in parts than whole but I do not want to dismantle this "work of art". (besides neither the specimen or gun chamber makes an attractive flowerpot).
Both have windowless EDS detectors and a backscatter detector. One has an optical microscope and a WDS system.
Dear Fellow MSA Listserver Members: I noticed the intense debate via email of the recent postings of "junk email" or "spam" on this List. Recently a report from a study sponsored by the Federal Trade Commission (FTC) concludes that there is basically no easy solution to this problem, although the FTC is working to solve it. The Consumer Protection Bureau of the FTC has asked that junk emails be forwarded to them at: uce-at-ftc.gov In the last year, they have prosecuted 5 businesses and warned 1000 more about spamming. This is probably the most effective way to handle the spam problem without one or only a few persons of this list saturating their time to fight against it. See also: http://www.ftc.gov/opa/9807/dozen.htm and http://www.ftc.gov/opa/9807/jbspam.713.htm for more info.
Hi there, are you in Arizona? Heck I would give them a home, besides if you have 2 of something that just means you have spare parts to keep things going with! Of course having electronics background does help.... I have an amr1000 I have yet to get opp. but part of that is my own lack of follow through. Since I have one of them It would be nice to have 2 of them! Therefore, that fact you have 2 is a great asset to the person that wishes to do own maint.
Coaters with only a mechanical pump do not give a satisfactory coating for studying morphology at reasonably high magnifications in SEM and TEM. They are however, quiet satisfactory for EDS or WDS analyses. It's not a matter of money, but most applications simply require a diffusion or turbo pump for carbon coating. Also, sputtering of carbon is unsatisfactory (VERY SLOW) and a carbon sputter head is no solution to carbon coating at all. Rotating the specimen during evaporation is effective for dealing with "difficult to coat" specimens. Disclaimer: ProSciTech distributes EMITECH equipment in SE Australasia only. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
-----Original Message-----
To all you printer users
Why do people prefer the Codonics printer when the Kodak seems to be 3/4 of the price and uses the same engine? I see more comments on the list about the Codonics than the Kodak. Is it just to have a networked printer or is the software better? Our need is primarily TEM (monochrome) but with maybe 10% colour, any comments from users with experience of either (or both) of these would be welcomed.
Thanks Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
I think I know the answer to this one, but does anybody out there have a = need / interest in the manuals described above? I am about to discard = them, but I'm finding hard to let go....
Going, going... gone?
James Wesley-Smith Electron Microscope Unit University of Natal, Durban South Africa
Which scope? If you have a Zeiss 410, write back and I'll pass along the secret :)
Tamara Howard CSHL
On Thu, 16 Jul 1998, Ping Li wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, can anyone there help me to put the scale bar onto a confocal image? } I saved } confocal image with scale bar (with overlay option), but once I open the } image } with any other imaging program, such as PhotoShop, the scale bar is not } on the } image any more. I wonder what I did wrong and how can I save the } confocal file } with scale bar as well as make the scale bar occurs when other imaging } softwares } other than LSM operating software are used. Your help will be greatly } appreciated. Thank you. } } Ping } pli-at-is.dal.ca } } } }
Two ways of doing this without burning the overlay into the image. 1. Perhaps the Zeiss software tells you the size of the field or the size of each pixel. Let's say each pixel is 0.285um and you want a 10um scale bar. Just divide to get the length in pixels to pop in with Photoshop. 2. Take an image of a micrometer slide.
-------------------------------------------- Michael Cammer email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 http://www.ca.aecom.yu.edu/aif/ --------------------------------------------
I have been trying to contact RMC at two different # and get a busy signal = for both. Can anyone give me their phone #, or e-mail address or any = explanation of what is going on? Thanks. Donna Wagahoff SIU School of Medicine Springfield,Il. 62794-1220 217-782-0898
Dr. Milos Kalab, a food scientist retired three years ago, has established a web site. Please visit following site if your are interested
http://www.cyberus.ca/~scimat/
He is still working hard in his laboratory.
Ann Fook Yang EM Unit Eastern Cereal and Oilseed Research Centre Agriculture and Agri-Food Canada 960 Carling Ave Central Experimental Farm Ottawa, Ontario Canada K1A 0C6
RMC 3450 So. Broadmont, Suite 100 Tucson, AZ 85713 Tel. (520) 903-9366 Fax (520) 903-0132 http://www.rmc-scientific.com/microtomes
Good luck, hope this helps!
Bob ***************************************** Robert (Bob) Chiovetti rchiovetti-at-aol.com E. Licht Company / 1-800-865-4248 Colorado/Utah/Arizona/Wyoming/ New Mexico/West Texas Representing Leica Since 1967 *****************************************
Two year ago we closed our EM Lab and I guess it is time I unsubscribe from this new group. (If some one would tell me how to.) Before I go I would like to share an essay I wrote just before we sold the EM and closed the lab.
To Whom It May Concern:
It will concern few, almost none, I suppose. Things change, seasons change. What were once considered major advances in the history of the human species, are regarded as important only to the past. I sit in a room which is cool, and dimly lit. There is a constant hum, so natural, that like breathing, it goes unnoticed. Today I hear this sound and try to embed it firmly in my memory. It is the sound of a voice I will not hear again. I focus on the sound, and struggle to imagine the sound of silence. All I can hear is the rhythm of the pump and the buzz of electronic. Most people see it as a huge piece of scientific equipment, just a Transmission Electron Microscope. It is not; it is a place. No No No. It is a magic carpet. It has taken me into the essence of life itself. I have crossed a cell membrane with more ease than sodium ions. I have followed an axon for microns, and watched as its microtubules dance in and out of view. I have observed actin and myosin locked in each other's embrace. I have beheld mitochondria in the thousands and found no two exactly the same. I have watched microvilli wave in rhythm like fields of wheat. I have looked at life (or rather the shadow of life) magnified 50,000 even 100,000 times. People have told me that this technology is obsolete, or they say that it is no longer cost effective for a small research department to support this type facility. Maybe it is their fault. Maybe it is the manufacturers fault, because they made service contracts too high. Maybe it is the government's fault, because it made money so tight. Maybe it is us for we have failed to see the direction science was turning and missed our chance to create the necessary techniques that would make this technology part of the wave of the future. Maybe it is my fault, because I did not show everyone all the things I have seen and places I have been. Astronauts have said that once you walk on the moon you no longer look at it the same way. NASA no longer goes to the moon and although it is still explores space, it seems to only be passing time. Like NASA, science will continue its exploration. Yes, just like NASA. I've not walked on the moon, but I have strolled where few people have tread. For those who have made my journeys possible I send my gratitude. It was definitely an "E" ride.
Thanks to those whom it did concern. To Mike, To Susan, To Ann.
Larry Hawkey, 1996 On the closing of the department's EM Lab.
Larry Hawkey hawkey-at-neuro.duke.edu Department of Neurobiology Duke University Box 3209 DUMC Durham, NC 27710 (919) 681-6425 fax (919)684-4431 http://www.duke.edu/~lah1
It is indeed a sad day when the accountants take over.
Managements general economic philosophy of restructuring, reorganising, reengineering, and retrenching unfortunately has no place in science and technology. Accountants (+snr man.) are reluctant to provide funding to reequip, repair, revitalise or replace.
LEO has a Cambridge S90 SEM for disposal (replaced by a new LEO 435). The SEM is at US Synthetics in Orem Utah. It was in working order when recently decommissioned, and has a BSD. Condition as seen, buyer collects, Any offers to Bill Neill billneill-at-csi.com please.
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
---------- Forwarded message ----------
Dear All,
We would like to purchase a rotatable (360 degree) X-Y stage for our Zeiss Axiovert 35M. Stages that fit the Axiovert 10 also fit this microscope. Even better yet, we would like to purchase just the X-Y part of this stage (Zeiss part number 45 35 60). If anyone has a used stage (or a new one), and would be interested in selling it, please contact me at Schwartz-at-rsbs.anu.edu.au
Thank you very much.
Sincerely,
Owen M. Schwartz
Owen M. Schwartz PhD Research School of Biological Sciences The Australian National University GPO Box 475 Canberra, ACT 2601 Australia
We are in the market for a new knifemaker. Those of you with newer model = knifemakers, please let me know how you rate their performance. Does anyone other that Leica make knifemakers? Thanks.
Donna Wagahoff SIU School of Medicine Springfield, Il. 62794-1220 217-782-0898 fax 217-524-3227
I've got a couple of users who would like to pursue Pre-embedment labling of plant tissue (trials with older knives and glass show excellent results) but are very concerned about potential damage to diamond knife edges. So I'm collecting opinions from experienced pre-embedment microtomists: are biological diamond knife edges basically 'safe' from 10-20 nm gold? Such that future biological sectioning will not suffer from any damage to the knife edge?
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
Try amazon.com on the web. They just found an out-of-print ultrastruct= ure book for me. It took them about 6 months, but they finally came up wit= h a slightly used copy. =
We are putting together a proposal to purchase a inverted microscope system for imaging of live cells. Here are a few general questions that I would love some feed back on:
1. Is Multi-Photon Imaging only available thru BioRad? We don't have the expertise to build one and can't afford the BioRad system.
2. If given the choice between a filter wheel or a monochromator for the excitation wavelengths, what is the better choice? I am concerned about the amount of light you can get through a monochromator. I figure I can always reduce the amoant of light from the filter wheel.
3. Is mercury vapor the best choice for the light source?
4. If you have an environmental chamber, is it also neccessary to have an objective heater or stage heater in order to have versatility in the kind of samples you can image?
5. Have you found that in some circumstances an upright system with a water immersion objective is more useful?
Opinions concerning any of these questions would be greatly appreciated. Thank you.
Bob Underwood Derm Imaging Center U of Washington Box 356524 Seattle WA. 98195-6524
Suggestions for adjustable height chairs (or modifications thereof) that minimize or eliminate fiber shedding and static charge would be appreciated. Thank you.
I am requesting this information for a colleague who is interested in automatic processors for paper and film. Has anyone used the Mohrpro 8 processor and what is their experience with it? You may reply to my email address directly if you wish.
Regarding the Codonics or Kodak printers....They are both built on the Kodak engine and look virtually identical. They also use the Kodak supplies for Dye-Sublimation printing and should give similar output. The difference is indeed in the software. The Codonics has completely rewritten the software and added many unique features which makes the printer much more versatile. I have found that the option to send multiple files to the printer and have them scaled and printed on one page a very desirable feature. That way I can send 4 SEM files at a time and have them printed as 4x5 with the cost of only 1 sheet of paper. That gives quick high quality copies similar to what we get with polaroid but cost is much less with digital capture and dye-sub printing (~$.50 per 4x5). The cost of the 1600 is not very different from the Kodak printer.
The Codonic NP-1660 has the added feature of being able to print on large pieces of sheet film and is very useful for medical imaging. It also will be able to produce high quality thermal prints at a projected cost of about $.50 per 8x10. There have been production problems with the thermal paper but my understanding (having just returned from the MSA meetings where I talked to Codonics representatives) is that they hope to have the media available within 2-3 months. Use of this would drop the cost of 4x5 study prints to a very low $.13/print and is what sold me on this printer. This is a UNIX printer that can be easily networked and image files can be sent postscript, via telnet or via the WWW. There are a huge range of compatable file types.
I have had some techinical problems with our Codonics printer but have had very good service....either another printer has been shipped within 24hrs in the case of hardware problems, or they diagnose and suggest fixes for software problems over the phone. I have been willing to cut them a bit of extra slack over the delay in delivery of the thermal paper because they have been very responsive in many other ways.
Since Kodak may have added features I am not aware of....do carefully check the spec's on both so you can choose the best for your purpose.
Debby Sherman ========================== Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Hi there,
... i am not quite sure, but I think that there has been a discussion about different dye-sub printers on this list-server... sorry for coming back to the same point, but here is my problem: we are planning to buy a high-end "photorealistic" printer. For certain reasons we tend to a Codonics ... Could anybody explain to me, what the advantage of the additionally implemented "direct thermal" technology (-} Codonics NP1660) is? The main task for our printer would B&W prints. The maximum percentage of colour prints is estimated to be in the region of 10 to 20%; pages/year appr.: 750..1000 ). It would be also very interesting to to have a comparison about the costs per page (Codonics NP1600 versus NP1660) including paper and ribbons, but excluding the investing costs for the printer itself.
Thank You very much in advance! Dipl.Ing.(FH) Gunnar Glasser, Elektronenmikroskopie
I don't know about shedding, but I have used a spray can of "Static Guard" or some such product. I think it was made by 3M. It worked very well during our winters when the inside humidity hit bottom. Of course our summers are so humid that we don't have static problems now.
At 11:34 AM 7/17/98 -0400, you wrote: } Suggestions for adjustable height chairs (or modifications thereof) that } minimize or eliminate fiber shedding and static charge would be } appreciated. Thank you. } } James Martin
I am neither a Kodak or Codonics user, but understand the following.=20
The Kodak undoubtedly has some processor built into it to handle the printing functions, and it is apparently adequate for many functions.=20
The Codonics incorporates a Sun Sparcstation as the smarts for their printer. That is apparently what gives it more flexibility, processing (not printing) speed, and network connectivity. Of course it comes at a cost. I do not know what the differences between the two Codonics models are.=20
--------- At 08:28 AM 7/16/98 +0100, you wrote: } Why do people prefer the Codonics printer when the Kodak seems to } be 3/4 of the price and uses the same engine? I see more comments on the=20 } list about the Codonics than the Kodak. Is it just to have a } networked printer or is the software better? Our need is primarily TEM } (monochrome) but with maybe 10% colour, any comments from users with } experience of either (or both) of these would be welcomed. } } Thanks } Ron
---------- and someone else wrote:
Hi there,
... i am not quite sure, but I think that there has been a discussion=20 about different dye-sub printers on this list-server... sorry for=20 coming back to the same point, but here is my problem: we are planning to buy a high-end "photorealistic" printer. For=20 certain reasons we tend to a Codonics ...=20 Could anybody explain to me, what the advantage of the additionally=20 implemented "direct thermal" technology (-} Codonics NP1660) is? The main task for our printer would B&W prints. The maximum=20 percentage of colour prints is estimated to be in the region of 10 to=20 20%; pages/year appr.: 750..1000 ). It would be also very interesting to to have a comparison about the=20 costs per page (Codonics NP1600 versus NP1660) including paper=20 and ribbons, but excluding the investing costs for the printer itself.=20
Thank You very much in advance! Dipl.Ing.(FH) Gunnar Glasser, Elektronenmikroskopie
MME offers customized on-site courses in all areas of microscopy. We have several consultants who work in this area. If we can be of help either visit our website at { {http://MME-Microscopy.com/education} or give me a call here in the office.
FEI Company currently has openings for experienced service engineers on SEM, TEM or FIB systems. Current areas of opportunity include Oregon, Idaho, California, Georgia, Virginia, and Texas. However, we are always looking for experienced engineers in any area.
If you would like to apply or refer an applicant, please contact me directly.
~~~~~~~~~~~~~~~~~~~~~~~~ Sarah A. Boydstun-Brown Sr. Human Resources Generalist FEI Company Phone (503)640-7556 Fax (503)726-2754 www.feic.com ~~~~~~~~~~~~~~~~~~~~~~~~
Dear Barna, } } I am trying to use a quantitative analysis program to analyze EDS spectra } collected with TEM. } I have 2 questions:
Several sessions of the recent MSA-MAS meeting were devoted to just these sorts of questions, and the proceedings are available on-line. In addition to any replies you receive, I suggest you look up those papers which appear relevant to you and contact the authors. This is just the reason why I make every effort to attend the meetings as often as I can; all the experts are within reach.
} (1) I have oxide inclusions in a diamond matrix . I think there are } absorption problems (Carbon absorbing oxygen). Is there anyway of getting } around it? I am using the "thin-section analysis" as an action. Should I } use " bulk sample analysis"? } The only ways "around it" are either to use appropriate standards (which contain a carbon matrix and a known amount of oxygen) or to use a correction to the oxygen intensity which accurately accounts for the absorp- tion. As was repeated several times at the meetings, this correction is usually the one with the greatest uncertainty. If your experimental con- ditions are such that the beam penetrates the specimen with little loss in energy and with little spread, then thin-section analysis is correct, and bulk sample analysis, which includes calculations for the production of x-rays by electrons which have deviated greatly from the incident dir- ection and lost a large fraction of their energy (among other effects), will give the wrong answer. The right procedure to use will always depend on the experimental conditions, and one must understand how the analysis soft- ware relates to those conditions in order to select the appropriate form of analysis.
} (2) I am using Nickel grid. So, the spectrum has Ni as an element in it. } How do I calculate the formula of the mineral? Do I perform the } quantitative analysis without the Ni? Or do I normalize the other elements } without taking into account that the Ni is present? } Is there anyway of determining if I do have Ni in my sample (or finding } out proportions of Ni coming from the grid and from the sample?) } If you were sure that your specimen did not contain nickel, you could subtract the nickel peak along with the continuum background and calculate the formula from what's left. You could put your sample on a different kind of grid and reanalyse it in order to determine whether it contains nickel--if a qualitative analysis shows no nickel, then all the signal is from the grid, and you can safely subtract it. You can also look at an area of the original grid which has the diamond matrix with no inclusions to see if the nickel peak is significantly reduced. In this case, be sure to check the continuum-to-nickel ratio. What you are trying to determine is whether the reduction in nickel signal is due to your moving to an area which contains less (zero) nickel, or whether the reduction is due to fewer electrons being scattered onto the grid bars (or, for that matter, pole pieces or any other nickel-containing part of your instrument) and producing a signal. This can become quite complicated if the average Z of the inclusions is very different from that of the matrix, and can vary depending on how large the inclusions are with respect to beam diameter and specimen thickness (i.e., what fraction of the analysis volume is inclusion). Good luck. Yours, Bill Tivol
I appreciate what Nestor has done and is doing with respect to the listserver and it is a lot of work. Basically, I think that he is aware of the problem and if he wants to tackle the problem, it should be up to him. Until he decides to do something, I am already deleting a lot of stuff that I'm not interested in without reading it. A few extra in a week doesn't bother me. Yes, some of it is offensive, but usually I know from the subject that it is spam and I delete it without reading. Sometimes it gets through. I try not to let it ruin my day as I delete it and forget it.
Essentially, let Nestor decide when enough is enough.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Dear all, As you will have noticed, there have been two recent ads posted to the list from non members (one for a sex site, and one for an email Marketing program). Whilst the vendors who subscribe to this group almost always behave in a responsible manner by sticking to the charter and not sending overtly commercial mailings and Nestor deals with the occasional abuse, we have no control about non list members who post onto the list.
I am aware that some other discussion groups use software that blocks postings from non list members. I suggest that it may be necessary for Nestor to configure the software that is used for this list to prevent posting from non list members.
What do other people think.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
} (1) Carbon will not appreciably absorb oxygen in your oxides. X-rays are } absorbed by elements heavier than the emitting element. The absorbtion } correction for oxygen in a carbon matrix, particularly in a thin film, will } be very small. Use the "thin-film" analysis mode.
X-rays are strongly absorbed when their energy is just above the energy for the transition from a filled state to a vacant state in the absorbing element. This topic is covered in Friedlander, et al. "Nuclear and Radiochemistry", and to quote, "...consider the K-alpha X rays of zinc (Z=30) which have an energy of 8.6 kV. The K absorption edges of Cu and Ni are 9.0 and 8.3 keV respectively. Therefore, nickel is a good absorber for zinc K-alpha X rays, and copper is not..." So your statement that x-rays are absorbed by heavier elements is not correct. The light-ele- ment folks would have memorized the O K-alpha and C K-edge energies (I haven't) and would know whether carbon absorption of O K-alpha is small or not. Yours, Bill Tivol
It depends on your microscope. If you use a coater system that does not have a clean vacuum system such as a turbo pump has, then you can get oil contamination on your sample that will show up in a high beam current density instrument such as a Field emission SEM. However, your coal samples could be a source of contamination even though you went with a more expensive system. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I am considering the purchase of an evaporative carbon coater for use on coal clinker specimens that have a history of bad charging under the SEM. The evaporative carbon coaters (carbon rod source) come in two types: high vacuum (10^-5 torr) and low vacuum (10^-2 torr). The high vacuum coaters (with a turbo pump) are significantly more expensive. Are they worth the extra cost?
Dear Catherine, } } Can anyone please advice on the proper disposal of uranyl acetate? } Thank you. } In New York small amounts of 1% UAc can be poured down the drain. Of course disposal of kg quantities of UAc is a different matter. Also, disposal regulations may vary from state to state, so it is best to check with your safety office. If they don't know, check with your state's environment department (ours is Environmental Conservation, but the name could be different in your state). Yours, Bill Tivol
Dear James, } } Suggestions for adjustable height chairs (or modifications thereof) that } minimize or eliminate fiber shedding and static charge would be } appreciated. Thank you. } Find one made of unuppolstered (sp?) metal, or rip out the uppol- stery and replace it with wood, or cover with aluminum foil (single-use). Yours, Bill Tivol
We've been cutting tissue attached to epoxy coated with 50 nm titanium for years. We keep a special diamond for it, which does need replacing more often than knives used for tissue only. But one can get excellent sections, thick and thin, and the knife does last for a while.
Lesley Weston.
On Fri, 17 Jul 1998 edelmare-at-casmail.muohio.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I've got a couple of users who would like to pursue Pre-embedment labling of plant } tissue (trials with older knives and glass show excellent results) but are very } concerned about potential damage to diamond knife edges. So I'm collecting opinions } from experienced pre-embedment microtomists: are biological diamond knife edges } basically 'safe' from 10-20 nm gold? Such that future biological sectioning will not } suffer from any damage to the knife edge? } } Thanks. } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "WE ARE MICROSOFT. } RESISTANCE IS FUTILE. } YOU WILL BE ASSIMILATED." }
Larry, Your sad commentary could not have reached me at a more pertinent time. Today, at this very hour, a group of faculty sit and discuss the future of our only general use EM facility here at Stanford University. We have tried, as a group of concerned microscopists, to let them know how important it is to keep our facility open. Maybe they need to go on a magic carpet ride to experience the beauty and importance of our "obsolete" technology. Won't there be another mutant that we need to look at soon? JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94022
Our apologies to anyone who is having trouble reaching us. It appears that the area code change to 520 is one culprit, the reassigning of the old number to someone in Phoenix is another.
The fact that US West keeps a seperate directory assistance database from AT&T is a third, moving across town to a new building is a fourth and having a record amount of activity is a fifth.
We have taken all the steps necessary to remedy this including notices to all directory assistance sources we know of and change of address and phone to all internet sources as well.
OUR NEW PHONE IS 520-903-9366, address: 3450 S Broadmont Tucson, AZ 85713
PLEASE NOTE: do not respond to my personal email address on this missive, I am traveling and use this only sporadically. We invite all to visit Tucson to see our new facility and new applications laboratory. We will give a tour of our production facility and even let you use a phone to prove that we have one. Apologies again to anyone that has been inconvenienced.
Regards. Steve Miller Director of Sales, North America
In response to the following posting from Catherine: ================================================================= Wonder if anyone going to the 14th ICEM in Mexico? Any idea of Cancun, such as safety and traveling around? ================================================================= My wife and I are "veterans" of many trips over the years to Cancun, both for holidays and also scientific meetings, and I believe we can speak from the perspective of first hand experience, the most recent trip being about 2 years ago.
The Mexican government designated some years ago certain areas called "zona turisticas" and Cancun was one of the first (if not the first). Basically, these special areas are run from a safety and hygiene standpoint virtually like you were in the USA. The US chain hotels (e.g. Sheraton, Marriott, Hyatt, etc.) seem to be run from a safety, security, and other standpoints as if you really were in the USA. I think the other hotels in the "zona turistica" are also run to that same standard, as are also the local outside-the-hotel restaurants with the zone. We always ask for "agua purificada" and we perhaps take a few other common sense precautions, but in all our many trips to Cancun neither of us has ever suffered any problems. Hotels outside the "zone" would not be necessarily operating to that kind of standard so further precautions should be taken. The one note of caution is this: Just as there is in parts of the US a problem with salmonella in eggs, there is also the problem in Mexico so you would want to make sure that any eggs in your diet were hard boiled or scrambled well done.
So far as "traveling around", we have taken both tours as well as gone off on our own in rented cars. We have never had any problems. Everything is very reasonably priced. We have found the Mexican people to be friendly, hospitable, and genuinely appreciative of those who visit their country, perhaps in part because that part of Mexico is so dependent on tourism. From a purely touristic point of view, there are far more things to see and do than one could possibly have time for doing. The top of my list would include a day trip to Chichen-Itza, and a day trip to the neighboring island of Cozumel. If you are a scuba diver, the "wall" dive on Palancar Reef, just off of Cozumel, is one of the world's greatest dives. However, you might be better off to take a hotel for a few days in Cozumel.
Like anywhere else in the world, common sense must prevail. For example, upon arrival at the airport in Cancun, there are numerous porters to carry your luggage to a waiting taxi. There are no trolleys as there are in most airports for doing this yourself. So if you have more luggage than you can carry on your own, be prepared to take advantage of the local porters. My advice is to convert some money at the currency window right there is in the luggage arrivals area, while you are waiting for the arrival of your luggage. The rate is usually better there than at airports in the US. You should be prepared to tip the porter US $.50 per bag (or the equivalent in pesos). You will need to purchase a ticket for the taxi (really a van holding 8-10 persons) to your hotel. Within the zona turistica, it is all the same taxi rate. The cost is per person and is about US $8-$9. A group of four or more, arriving at the same time and going to the same hotel can get a better deal by reserving an entire taxi. So you should convert enough money to end up with enough pesos in order to be able to purchase your taxi tickets and give the porter a tip and also the driver a tip. If you present for payment for the taxi tickets US dollars, the rate you are given (at least that was the case in the past) is pretty crummy. Therefore the reason to get some pesos. When leaving the airport area, for the taxi ticket stand or anywhere for that matter, I would recommend keeping a close eye on your belongings. I would make this same admonition for someone arriving at JFK in NYC, but this is just common sense.
If you should arrive and one or more of your bags does not arrive, then you want to file the report with the airline before leaving the luggage area and before going through customs. If you are connecting from a different airline to make your final flight to Cancun, make sure when you check in for the Cancun flight you present your bag tags from the previous flight so make sure your bags are in fact transferred onto the Cancun flight. Other wise, with security being what it is, you could end up being separated from your checked luggage.
} From all that I have been able to learn, it really does sound like this ICEM is going to be the best ever. When I attended the MICRO 98 meeting in London a few weeks ago, the "word" was that "everyone" is going. However, there are still luxury hotel rooms available at reasonable rates so if you have not yet made your decision, now is the time to decide! One other note: Don't worry if a hotel is one or more Km from the convention center. A local bus traverses the main road along the hotel strip every few minutes and the cost per ride is roughly US$.25 , so don't let the "distance from the convention center" deter you from a particular hotel (so long as it is along the hotel strip). In any case, riding these local buses is a part of the Cancun experience you would not want to miss anyhow.
Chuck
PS: For prospective exhibitors, I am told there are still some booths (exhibit stands) available.
Disclaimer: These are my own personal opinions and recommendations and might not necessarly reflect the views of the organizers.
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
In response to the following posting from Catherine: ================================================================= Wonder if anyone going to the 14th ICEM in Mexico? Any idea of Cancun, such as safety and traveling around? ================================================================= My wife and I are "veterans" of many trips over the years to Cancun, both for holidays and also scientific meetings, and I believe we can speak from the perspective of first hand experience, the most recent trip being about 2 years ago.
The Mexican government designated some years ago certain areas called "zona turisticas" and Cancun was one of the first (if not the first). Basically, these special areas are run from a safety and hygiene standpoint virtually like you were in the USA. The US chain hotels (e.g. Sheraton, Marriott, Hyatt, etc.) seem to be run from a safety, security, and other standpoints as if you really were in the USA. I think the other hotels in the "zona turistica" are also run to that same standard, as are also the local outside-the-hotel restaurants with the zone. We always ask for "agua purificada" and we perhaps take a few other common sense precautions, but in all our many trips to Cancun neither of us has ever suffered any problems. Hotels outside the "zone" would not be necessarily operating to that kind of standard so further precautions should be taken. The one note of caution is this: Just as there is in parts of the US a problem with salmonella in eggs, there is also the problem in Mexico so you would want to make sure that any eggs in your diet were hard boiled or scrambled well done.
So far as "traveling around", we have taken both tours as well as gone off on our own in rented cars. We have never had any problems. Everything is very reasonably priced. We have found the Mexican people to be friendly, hospitable, and genuinely appreciative of those who visit their country,
perhaps in part because that part of Mexico is so dependent on tourism. } From a purely touristic point of view, there are far more things to see and do than one could possibly have time for doing. The top of my list would include a day trip to Chichen-Itza, and a day trip to the neighboring island of Cozumel. If you are a scuba diver, the "wall" dive on Palancar Reef, just off of Cozumel, is one of the world's greatest dives. However, you might be better off to take a hotel for a few days in Cozumel.
Like anywhere else in the world, common sense must prevail. For example, upon arrival at the airport in Cancun, there are numerous porters to carry your luggage to a waiting taxi. There are no trolleys as there are in most airports for doing this yourself. So if you have more luggage than you can carry on your own, be prepared to take advantage of the local porters. My advice is to convert some money at the currency window right there is in the luggage arrivals area, while you are waiting for the arrival of your luggage. The rate is usually better there than at airports in the US. You should be prepared to tip the porter US $.50 per bag (or the equivalent in pesos). You will need to purchase a ticket for the taxi (really a van holding 8-10 persons) to your hotel. Within the zona turistica, it is all the same taxi rate. The cost is per person and is about US $8-$9. A group of four or more, arriving at the same time and going to the same hotel can get a better deal by reserving an entire taxi. So you should convert enough money to end up with enough pesos in order to be able to purchase your taxi tickets and give the porter a tip and also the driver a tip. If you present for payment for the taxi tickets US dollars, the rate you are given (at least that was the case in the past) is pretty crummy. Therefore the reason to get some pesos. When leaving the airport area, for the taxi ticket stand or anywhere for that matter, I would recommend keeping a close eye on your belongings. I would make this same admonition for someone arriving at JFK in NYC, but this is just common sense.
If you should arrive and one or more of your bags does not arrive, then you want to file the report with the airline before leaving the luggage area and before going through customs. If you are connecting from a different airline to make your final flight to Cancun, make sure when you check in for the Cancun flight you present your bag tags from the previous flight so make sure your bags are in fact transferred onto the Cancun flight. Other wise, with security being what it is, you could end up being separated from your checked luggage.
} From all that I have been able to learn, it really does sound like this ICEM is going to be the best ever. When I attended the MICRO 98 meeting in London a few weeks ago, the "word" was that "everyone" is going. However, there are still luxury hotel rooms available at reasonable rates so if you have not yet made your decision, now is the time to decide! One other note: Don't worry if a hotel is one or more Km from the convention center. A local bus traverses the main road along the hotel strip every few minutes and the cost per ride is roughly US$.25 , so don't let the "distance from the convention center" deter you from a particular hotel (so long as it is along the hotel strip). In any case, riding these local buses is a part of the Cancun experience you would not want to miss anyhow.
Chuck
PS: For prospective exhibitors, I am told there are still some booths (exhibit stands) available.
Disclaimer: These are my own personal opinions and recommendations and might not necessarly reflect the views of the organizers.
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
FALL 1998 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-V2)
NASSAU COMMUNITY COLLEGE
A fifteen week, fall 1998 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered ONE EVENING PER WEEK, Thursdays, starting at 5:30pm. Classes will begin on Sept. 3 and end on Dec. 10, 1998.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 per credit.
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
For those without www access, the catalog description is specified below. If you have further questions, you should e-mail me directly at the address below.
Interested individuals should register early (by Aug. 10) since the course is limited to a total enrollment of ten (10) students.
Questions regarding the actual registration process can be directed to our registrar at (516) 572-7355. ________________________________________________________________________________
CATALOG DESCRIPTION BIO 221: Transmission Electron Microscopy -- 4 credits Prerequisites: BIO 109-110 or equivalent, CHE 151-152 or equivalent. An introduction to the basic principles of transmission electron microscopy including tissue preparation, microscope (TEM) operation, black & white photography, and micrograph interpretation. The entire laboratory is devoted to the development of skills and preparative techniques involved with the operation of an actual transmission electron microscope. (3 lecture, 3 laboratory hours). Laboratory fee applies. ________________________________________________________________________________
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
FALL 1998 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-V2)
NASSAU COMMUNITY COLLEGE
A fifteen week, fall 1998 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered ONE EVENING PER WEEK, Thursdays, starting at 5:30pm. Classes will begin on Sept. 3 and end on Dec. 10, 1998.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 per credit.
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
For those without www access, the catalog description is specified below. If you have further questions, you should e-mail me directly at the address below.
Interested individuals should register early (by Aug. 10) since the course is limited to a total enrollment of ten (10) students.
Questions regarding the actual registration process can be directed to our registrar at (516) 572-7355. ________________________________________________________________________________
CATALOG DESCRIPTION BIO 221: Transmission Electron Microscopy -- 4 credits Prerequisites: BIO 109-110 or equivalent, CHE 151-152 or equivalent. An introduction to the basic principles of transmission electron microscopy including tissue preparation, microscope (TEM) operation, black & white photography, and micrograph interpretation. The entire laboratory is devoted to the development of skills and preparative techniques involved with the operation of an actual transmission electron microscope. (3 lecture, 3 laboratory hours). Laboratory fee applies. ________________________________________________________________________________
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
I am writing to request some help please. I am a Chiropractic Physician who has taken two training courses in Enderleinian Darkfield Microscopy. I am searching for a used microscope to use for that purpose. I would like your opinion comparing the quality of the Zeiss Universal to a Leitz Orthoplan. Also, is IC optics a significant help with darkfield? Is the image from an Axioskop any better than that obtained from a Universal or ORthoplan? Finally, which type of objectives - plan Achromat. plan-Neoflour, or Plan apochromat would suffice/be recommended? Thank you very much.
Numerous micrographs of bacteria and other specimens can be found in Ruska's book: Ernst Ruska. 1980. The early development of electron lenses and electron microscopy. Translated by Thomas Mulvey. S. Hirzel Verlag (Stuttgart). ISBN 3-7776-0364-3.
A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
-------------------------------------
On Mon, 20 Jul 1998, frank booy wrote:
} I remember seeing an image of a bacterium imaged in/by Ruska's } first microscope. } Can anyone tell me where this is to be found please? } Thanks! } } Frank Booy }
I am currently looking into a new Field Emissions Auger for our lab. I know the general differences between the Cylindrical Mirror and the Hemispherical analysers but has anyone performed a recent comparison. I run a multipurpose system doing powders, fracture surfaces, polished surfaces and anything in between. If anyone has data that they can share with me I would really appreciate it.
Thank you Tamara E. Bloomer Assistant Scientist Ames Laboratory 137 Wilhelm Hall Ames, IA 50011 (515) 294-2564
Donna Wagahoff asked: } We are in the market for a new knifemaker. Those of you with newer model } knifemakers, please let me know how you rate their performance. } Does anyone other that Leica make knifemakers? Thanks.
We (Energy Beam Sciences) manufacture both a triangular and a "Ralph" glass knifemaker. The triangular knifemaker is the descendent of the old DuPont-Sorvall knifemaker, and is designed to cut 10mm and 12mm knives (as opposed to the Leica, which is designed for 6.4mm glass). RMC also make a knifemaker, and there is another one, made in the UK, which is distributed through Agar there.
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
First: regarding the choice between Zeiss and Leitz
The best suggestion for your application is that you try your specimen with the microscope you intend to purchase, to see which does the best job for you. I am not familiar with the term "Enderleinian" with respect to Darkfield microscopy. If you can provide a description, I may be able to comment more completely on which optics would perform best.
Secondly: regarding "IC" optics
This nomenclature refers to a different contrast technique, differential interference contrast (DIC), often called "Nomarski" (Dr. Georges Nomarski developed a method for cutting the necessary prisms which enabled the technique to become commercially viable). Since DIC is based on polarized light, it requires strain free optics; the engraving IC indicates that the optics are suitable for this technique. Making the assumption that you are working in transmitted light, a complete DIC system would require a set of objectives fitted with polarizers and DIC prisms (some systems have a single, common prism while others have a separate prism/polarizer set for each objective) as well as a matching condenser (often a turret, if you are going to do DIC with more than one objective), also fitted with polarizers and prisms.=20
These two techniques work on very different aspects of the sample:
In Darkfield, the light approaching the sample comes in at such a high angle that it cannot be collected by the objective. As a result, the background is dark. However, any feature in the sample (little grains, strings, pits, scrathes, dust) which can scatter light will do so. The scattered light is collected by the objective and used to form the image. The result: bright objects against a dark background. Since scatter is highly wavelength dependent, darkfield can also cause is extraneous color.
DIC is a bit too complex to explain completely here but basically, it detects gradients (slopes or changes)in the sample. The gradients can come from either changes in topography or changes in refractive index (ex: due to changes in concentration, gelation, etc.). In the most sensitive DIC settings, the background is usually pearly gray; one slope will be bright and the slope on the opposite side of the feature will be dark. Since you mind interprets these bright/dark cues in terms of differences in height, DIC images often appear to have increased structural or 3-dimensional information; they need to be interpreted very carefully. Sometimes adding an internal standard (ex: tiny glass beads) is helpful in understanding what is really happening. A second artifact comes from materials which respond to polarized light. Typically, they will "scramble" the DIC information. What you will see will be beautiful Pol colors, but not the true gradient information from the DIC. To test, just compare one side of the feature to the other. If the feature shows the same response on all sides (as opposed to one side being bright, the other being dark), it is probably responding to the Pol component of the system, not the DIC components.
For more on all these topics, might I suggest our book "Optimizing Light Microscopy for Biological and Clinical Laboratories"? Details are available at our website:
Thank you to those who responded to my inquiry after clean-room chairs.
We located several fitting the description in the Lab Safety Supply catalog (800-356-0783) made by a company called BioFit. The chair seats are vinyl (and look comfortable) and dissipate static by means of a brass chain fitted to the bottom of the chair, which drags on the ground. The difference between this chair and regular vinyl chairs appears to be the brass grounding chain.
No doubt other vendors carry similar products, and I have no commercial interest (of which I know) in Lab Safety Supply Co.
Thanks everyone for helping me try and build or get a microscope. My feelings are that it is better and cheaper to buy a microscope, ie - 600x from India or China for $70, (one of the plans I saw was to make a 100x for $100, but it seems outdated) but I still want to make my own eventually someday. Could someone direct me to some sources for acquiring these microscopes? Perhaps an online source or catalog. I want to use the scope to view slides with a light source from underneath, and also be able to view the surface of small objects (like within an inch in heigth or bigger).
We do not believe you would need a turbo system. Ladd sells a carbon coater (diffusion pump) that is a high vacuum system (10 to the minus 6 or 7) and we believe this would be enough for you. We could always add a turbo pump option that would add about $6,000 to the base price, which I would be happy to quote you if you are interested.
John Arnott Chairman --
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
Rick Felten 07/20/98 04:25 PM I love my Stylus Color 800 and need another printer. Has anyone compared a Epson Stylus Photo 700 to a Stylus Color 800 for printing high resolution black and white images? Ric Thanks
What do you mean by IC optics: DIC (Differential Interference Contrast or Nomarski) or ICS (Infinity Corrected System (Zeiss's infinity optics)?
Infinity correction will help if you are putting anything into the light path between the objective and the Telen (projection) lens (which focusses the light rays properly for the ocular lens.
Generally apochromatic lenses have higher numerical apertures and hence higher resolving capabilities than neofluars or achromatic lenses.
As to which microscope is better for your darkfield, I think you just have to try out the different microscopes for your particular application. This may be difficult if you're trying to buy a used one, but some of the microscope companies do sell used ones.
Matt Schibler
} ---------- } From: Stephen[SMTP:smd-at-capecod.net] } Sent: Sunday, July 19, 1998 3:37 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Zeiss vs. Leitz + } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Good day to everyone who is reading this, } } I am writing to request some help please. I am a Chiropractic } Physician } who has taken two training courses in Enderleinian Darkfield } Microscopy. I } am searching for a used microscope to use for that purpose. I would } like } your opinion comparing the quality of the Zeiss Universal to a Leitz } Orthoplan. Also, is IC optics a significant help with darkfield? Is } the } image from an Axioskop any better than that obtained from a Universal } or } ORthoplan? Finally, which type of objectives - plan Achromat. } plan-Neoflour, or Plan apochromat would suffice/be recommended? } Thank you very much. } } } } Stephen M. Driscoll }
May I suggest an HP 890C ($399). I've really been impressed with the quality when using the both the special paper and normal paper. I bought the cheaper version of this printer, the 722C ($299) for home and have been happy with it also. Both of these printers have the PhotoREt II feature and the Kodak Image Enhancements. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Rick Felten 07/20/98 04:25 PM I love my Stylus Color 800 and need another printer. Has anyone compared a Epson Stylus Photo 700 to a Stylus Color 800 for printing high resolution black and white images? Ric Thanks
I was wondering if anyone could help me analyse some results I have obtained using acridine orange. I am using rat uterus in my experiments, and attempting to detect apoptosis in the tissue. My main problem is interpreting the changes in DNA and RNA content in the tissue; so if there are any experts out there who could possibly give me some assistance, that would be great.
Thanks
Richard
Dr Richard Stump Rm 221 Anatomy and Histology Anderson Stuart Bldg. F13 University of Sydney NSW 2006
} } Thanks everyone for helping me try and build or get a microscope. My feelings } are that it is better and cheaper to buy a microscope, ie - 600x from India or } China for $70, (one of the plans I saw was to make a 100x for $100, but it } seems outdated) but I still want to make my own eventually someday. Could } someone direct me to some sources for acquiring these microscopes? Perhaps an } online source or catalog. I want to use the scope to view slides with a light } source from underneath, and also be able to view the surface of small objects } (like within an inch in heigth or bigger). } } Danny -
Here's a list of sources of compound scopes in your price range. a 3 objective monocular with condenser will cost at least $150. You'll find several books for adult hobbiests in the Project MICRO bibliography (address below).
3-OBJECTIVE MONOCULAR COMPOUND SCOPES
There is a much greater selection in this category; follow the advice in "Microscopic Explorations", and shop around. Every major school supplier offers a selection. Here are a few possibilities: Carolina Biological Supply 800-334-5551 Edmund Scientific 800-728-6999 Educational Teaching Aids 800-445-5985 Frey Scientific 800-225-FREY Insights Visual Productions 800-942-0528 Lakeshore Learning Materials 800-421-5354 Nasco 800-558-9595 Sargent-Welch 800-727-4368 Southern Precision Instruments (dealer) 800-678-7768
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I am posting this message for another microscopist. Your reply direct to his email or the listserver will be appreciated. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes *********************** www.proscitech.com.au *****
} I am looking for a staining method for looking at slime } deposits from } paper mills and biofilms in general using light microscopy. } I came } across some methods that stain bacterial capsules but none } that mentions } biofilms or slime layers (particularly levulose polymers). } } Bill van Eijk } email: whvanei-at-ibm.net
Dear Barbara, } } The chair at my microscopy bench has a naugahyde cover. Would that } help? } If one has the spiffy new synthetic-fabric lab coats (rather than the old-fashioned cotton ones), naugahyde is not a solution to the static- electricity problem. One could almost use that combination for an emer- gency HT source ;-). Yours, Bill Tivol
I'm sorry it's taken so long to summarize the responses I received to my query on how long to store tissues in glutaraldehyde fixative. The results are about even regarding whether or not to do it. Some people strongly believe that it is perfectly fine to store tissues for extended periods of time in glutaraldehyde--a much better way to store tissues than in buffer--even with preservatives and anti-.fungals. Several laboratories responded that they store tissues in glutaraldehyde all the time with no adverse effects. One laboratory mentioned that on occasion, they do see some membrane whorling in their tissues following long storage periods, but it is minor. One laboratory described very negative effects of extended storage in glutaraldehyde, such as loss of density and contrast. Another individual mentioned a paper written a few years ago that dealt with different formulations of fixatives and found that storage in a fixative called 4F1G (4% formaldehyde/1% glutaraldehyde in PO4 buffer) over a 5 year period showed no adverse effects, but storage in higher concentrations of glutaraldehyde may cause some artifacts. One laboratory reported that extended fixation in glutaraldehyde makes the tissue difficult to section due to hardening of the tissue. Additionally, they report occasional leaching of detail and less intense staining. Finally, an elegant paper from the South African Journal of Botany ( Coetzee J & van der Merwe C F - 1986. The influence of processing protocol on the ultrastructure of bean leaf cells. SA J Bot 52 (2) 95-99) describes several fixation schedules, vaying fixation times, buffers and periods in buffer wash. The authors suggest that maximum fixation times in Na-Cacodylate should not exceed 16 hr, phosphate buffers should not exceed 2 days, and PIPES-buffered glutaraldehyde fixation should not exceed 4 hours. The most accepatble fixations with all the bufferes were obtained with one hour fixation schedules. Those of us who have worked with plant material are well aware that plant material is much less forgiving than animal tissue, which may account for the varying accounts and opinions. The bottom line from just about everybody, however, is that it is best simply to not store tissues at all and immediately process them up to the block stage (though we all know that this is not always possible). I hope this information helps those who requested a summary. I greatly appreciate everybody's input (and it was great to hear from some old friends again!). Thanks again! Cheers, Laura
Laura M. Patrone, Ph.D. Wyeth-Ayerst Research Biomedical Imaging 641 Ridge Road Chazy, NY 12921 (518) 846-6318 e-mail: patronl-at-war.wyeth.com
Thank you very much for taking the time to put that info out there. I would be interested in what kind of weather to expect also if anyone has that kind of information.
Hi all, I am organizing the Materials Science Tutorials for the 1999 M&M Meeting and earlier I had asked for suggestions on topics from MAS and MSA members and other possible attendees of the conference. Below is a list of what I received in the way of suggestions. I am asking those of you who have the time to vote for up to three of the subjects. This will help me make the final decision on the topics for next year. If you have other suggestions or comments please let me know.
} 1. "How about GIF and perhaps PEELS?" } } 2. Defect recognition and analysis in crystalline materials. } } 3. Accessing and using on-line crystallogrphic databases } } 4. SEM Techniques } } 5. Optimizing the microscope for XEDS and WDS detection } } 6. XEDS imaging and interpretation, the right ways and the wrong ways. } } 7. Automation and Remote Control } } 8. More sample preparation (cross section of ALL methods) } } 9. Cross section samples with the FIB & FIB in general. } } 10. Spectrum imaging
I should note that FIB was a very popular choice and I have pretty much decided to do that one anyway, but I would still like to hear some yeahs or nays.
Thanks.
John M.
Note new Area Code (734)
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Does anybody out there know how to get Leica to respond? We bought a camera system for one of our microscopes and the parts are not compatable. We have been trying to talk to our Rep. and he will not return the phone calls. I also sent an email to Leica with no response. Any ideas will be appreciated.
Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
} 9. Cross section samples with the FIB & FIB in general.
} 10. Spectrum imaging
and
EBSP
Thanks for your efforts in organizing these tutorials.
Scott
F. Scott Miller Electron Microscopy Lab smiller-at-umr.edu University of Missouri-Rolla 223 McNutt Hall voice: 573 341 4727 Rolla, MO 65409 USA fax: 573 341 6934
Can anyone tell me for sure that Tannic acid is Ok to be used in Phos buffer (at about 6.9 pH)? (for fixation after glutaraldehyde). We have always used Cac bfr, but cannot in this instance. If you just answer "yes" or "no", it is good enough. Thanks, Hildy
While on assignment, I supported a lab which did a lot of work with biofilms which formed everywhere from cooling towers for power plants to beer facilities. They used DiI (UV excitation).
For the best reference for all sorts of stains, dyes, and probes:
Molecular Probes Handbook of Fluorescent Probes & Research Chemicals,
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Does anyone have experience with the nanoplast resin? I would like to know if it possess a column contamination problem when used in the TEM or SEM. Also, are the results equal to or better than conventional resins or other resins that can tolerate some water remaining in the specimen material prior to infiltration. The intended use is for isolated starch granules. These granules are often hydrated during preparation or when used in food products. When hydrated, they are much softer and somewhat swollen so resins may be able to penetrate easier. Also, it may be of interest to compare the hydrated state with the dried morphology.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
by news.mtu.edu (8.8.8/8.8.8) with ESMTP id PAA25397; Tue, 21 Jul 1998 15:42:43 -0400 (EDT) Received: from mmserver.mm.mtu.edu (mmserver.mm.mtu.edu [141.219.66.30]) by mtu.edu (8.8.8/8.8.8) with ESMTP id PAA04040; Tue, 21 Jul 1998 15:42:43 -0400 (EDT) Received: from backscatter (backscatter.my.mtu.edu [141.219.67.144]) by mmserver.mm.mtu.edu (8.8.7/8.8.7/mturelay-1.2) with SMTP id PAA25134; Tue, 21 Jul 1998 15:42:41 -0400 (EDT) X-Authentication-Warning: mmserver.mm.mtu.edu: Host backscatter.my.mtu.edu [141.219.67.144] claimed to be backscatter Message-Id: {3.0.5.32.19980721153734.0097d360-at-mmserver.mm.mtu.edu} X-Sender: opmills-at-mmserver.mm.mtu.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.5 (32)
A colleague at Tulane University asked that I submit this to the list. Please do not reply to me.
Cheers, Owen
} LAB SUPERVISOR (TEM/SEM) } } } Tulane University's Coordinated Instrumentation Facility seeks an } individual to create its EM facility consisting of 2 SEM's (Amray 1600T, } Jeol 820) and TEM (Philips 410). The successful candidate will possess } experience in operating within a multi-user environment serving biological } and physical sciences clientele. Experience with basic } biological/materials techniques is required including, histochemistry, } ultramicrotomy and thin foil preparation, electron diffraction, x-ray } microanalysis (EDS & WDS), photographic and darkroom procedures, digital } imagery practices. Experience is desired in the following: repair of } electron microscopes, high vacuum systems and accessory instrumentation and } basic computer skills (word process, spreadsheet, digital image } manipulation, HTML). B.S. physical/engineering required, M.S. preferred. } } Qualified applicants submit complete packet of cover letter and resume in } our office on or before August 31, 1998 to: } } Tulane University } Human Resources } Collins C. Diboll Complex } New Orleans, LA 70118 } } Tulane University is an AA/EOE. } }
Presuming a hurricane does not hit the week of the ICEM meeting, there are two words which accurately describe the weather in Cancun the first week of September: Steam Bath.
Dress (or undress) accordingly...
Larry
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Responding to the message of {n1311092614.19691-at-btny.purdue.edu} from "Microscopy Center" {emcenter-at-btny.purdue.edu} :
I have one grad student in my lab who is using it for gold labeling studies on algae. He is using it because he doesn't have to completely dehydrate the cells in organic solvents before infiltrating them.
I don't know if there is any problem with contamination of TEM column, at least none that I'm aware of........yet.
One important clue when using this resin, is blocks tend to come out VERY hard if you use the recommended amount of catalyst B-52, such that you can't even trim it. We use 1/4 the amount specified in the instructions that come with the resin. So advise you try a few curring runs of blocks with no sample just to work out the amount of catalyst to use such that you get blocks soft enough to trim and cut.
A thread on nanoplast ran on this listserver in March. I compiled them into a file and will send it to you off-line as an attachment.
Good luck!
Gib
} Does anyone have experience with the nanoplast resin? I would like to know } if it possess a column contamination problem when used in the TEM or SEM. } Also, are the results equal to or better than conventional resins or other } resins that can tolerate some water remaining in the specimen material prior } to infiltration. } The intended use is for isolated starch granules. These granules are } often hydrated during preparation or when used in food products. When } hydrated, they are much softer and somewhat swollen so resins may be able to } penetrate easier. Also, it may be of interest to compare the hydrated state } with the dried morphology. } } Debby Sherman, Manager Phone: 765-494-6666 } Microscopy Center in Agriculture FAX: 765-494-5896 } Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu } Purdue University or: emcenter-at-btny.purdue.edu } 1057 Whistler Building } West Lafayette, IN 47907-1057 } } } .
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
May I correct Steven Slap's comment on the Leica knifemaker - +ACI-as opposed to the Leica, which is designed for 6.4mm glass+ACI-.
The Leica EM KMR2 is designed to produce glass knives using a balanced break technique from 6.4, 8.0 and 10.0 mm glass. If you would like more details - contact the friendly folks at Leica.
Philip Hyam Product and Marketing Manager - Sample Preparation Leica Microsystems Canada
I've had similar experiences in my area. You might try to find your rep's boss's name, or try the management chain in headquarters at Deerfield IL (800-248-0123).
Leonard R. Corwin Fort Dodge Animal Health Princeton, NJ 08543-0400
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Does anybody out there know how to get Leica to respond? We bought a camera system for one of our microscopes and the parts are not compatable. We have been trying to talk to our Rep. and he will not return the phone calls. I also sent an email to Leica with no response. Any ideas will be appreciated.
Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
by imo21.mx.aol.com (IMOv14_b1.1) id RYPZa12330; Tue, 21 Jul 1998 17:27:06 -0400 (EDT) Message-ID: {34b297c0.35b507ac-at-aol.com}
In a message dated 98-07-21 12:18:37 EDT, mmr7001-at-axe.humboldt.edu writes:
{ { Does anybody out there know how to get Leica to respond? We bought a camera system for one of our microscopes and the parts are not compatable. } }
Marty,
I agree with Leonard Corwin, Leica Customer Service at (800) 248-0123 should be able to help you. I don't know which e-mail address or web page you tried, but your request may be floating in cyberspace somewhere in Germany.
Did you purchase from Leica directly or through one of their regional dealers? Territories and reps change, and it's possible your area has been reassigned. Anyway, just give Customer Service your Zip code, and they can take it from there.
Hope this helps! Bob ***************************************** Robert (Bob) Chiovetti rchiovetti-at-aol.com E. Licht Company / 1-800-865-4248 Colorado/Utah/Wyoming/Arizona/ New Mexico/West Texas Representing Leica Since 1967 *****************************************
I have used LR Gold for embedding samples destined for EM immunocytochemistry on many occassions with no trouble. In my latest embedding, the blocks are not behaving in a typical fashion. Everytime I go to section them, they pull the water out of the boat and prevent thin sectioning. I can cut 0.5 um LM sections with effort and the sections look okay. Has anybody else had this experience and, more importantly, do you know how to get around it? TIA, tom phillips
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
} From all the fixations I did many years ago with Tannic acid after glutaraldehyde, my short answer is
YES.
P.S.: Try to use the lightest molecular weight mixture of tannic acids you can find. Ones from Turkish nutgalls, not Chinese nutgalls. Mallinkrodt #1763 or #1764 are what I used.
} ---------- } From: HILDEGARD CROWLEY[SMTP:hcrowley-at-du.edu] } Sent: Tuesday, July 21, 1998 9:43 AM } To: postmessage } Subject: Tannic acid+Phos bfr????? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Hi, } } Can anyone tell me for sure that Tannic acid is Ok to be used in Phos } buffer (at about 6.9 pH)? (for fixation after glutaraldehyde). We have } always used Cac bfr, but cannot in this } instance. If you just answer "yes" or "no", it is good enough. } Thanks, } Hildy }
Tom, you may have already considered these solutions. They apply to = sectioning: 1. maintain a lower than normal water level in the boat, so the level is = slightly concave but maintains contact with the edge of the knife; and = 2. use a faster cutting speed. These are effective in reducing that = notorious water "pulling" by the LR's. Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
} The sample is embedded in a Spurr's resin. I am currently using an } Iodine solution but I was wondering if there was anything better? } } Robert } } Robert.Dickson-at-kcl.fi
You should be able to stain Spurr's with PAS provided the 1% periodic acid step is long enough, e.g. 30 minutes. If you want the complete procedure get back to me.
The staining should be a lot more intense than with Iodine, but other cell components may also stain up. Presumably you are looking at plant tissue? Some cell wall polysaccharides are likely to stain, also some phenolics. Nuclei may also stain - see Litwin, JA & Kasprzyk, JM (1974) Acta Histochemica, vol 50, pp 222-227 "Some remarks on the PAS reaction in semi-thin sections of Epon-embedded tissues". If your tissue was embedded in methacrylate resin or similar you could do a control of sorts by digesting the section with amylase before staining, but this may not work on Spurr's.
O'Brien and McCully recommend aniline blue black as a counterstain: 1% aniline blue black in 7% acetic acid for 10 min at 50 deg.C, rinse in 7% acetic acid. (Yes, I've got a copy of their out-of-print book, and it's not for sale).
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
Philip Hyam wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } May I correct Steven Slap's comment on the Leica knifemaker - +ACI-as opposed to the } Leica, which is designed for 6.4mm glass+ACI-. } } The Leica EM KMR2 is designed to produce glass knives using a balanced break technique } from 6.4, 8.0 and 10.0 mm glass. If you would like more details - contact the } friendly folks at Leica. } } Philip Hyam } Product and Marketing Manager - Sample Preparation } Leica Microsystems Canada
Right Philip! But then again, if you are on a limited budget, the "old" LKB Knife Makers still do a great job for 6,4 mm glass. They can be reconditioned (by us) at a very reasonable cost and reconditioned units are available. Markus F. Meyenhofer Microscopy Labs micro-at-mail.superlink.net
I really dislike "me too" postings, but I have a similar complaint to = Marty's. I would like to suggest to any Leica personnel out there to = make your Head office aware of the need for greater accessibility of = your applications dept. This means no disregard to regional offices, but = it is often helpful to talk to people closely involved in the = development of a particular instrument.
James Wesley-Smith Electron Microscope Unit University of Natal,=20 Durban, South Africa
----------
Does anybody out there know how to get Leica to respond? We bought a camera system for one of our microscopes and the parts are not = compatable. We have been trying to talk to our Rep. and he will not return the phone calls. I also sent an email to Leica with no response. Any ideas will = be appreciated.
Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
I have just heard from Prof. Sitte that, following his retirement, this year's workshops will be the last. So, having been a speaker in them since 1986, I thought I would say a few words.
For those who do not know, the workshops have been running in Seefeld (Tirol) since 1973 and, before that, in the University of Homburg, Germany. Over 2,000 biomedical and materials scientists and technicians from around the world have participated in them.
The workshops are renowned for their technical content, social intercourse, gastronomic experience, cultural interest and scenic beauty (being on a high plateau in the Alps). Somehow, Sitte has perfected the art of being the gracious host who combines long hours of learning with enjoyment.
The courses are taught by various experts in their fields: some are the developers of ultramicrotomy and cryo-equipment, some are immuno experts and others have made fundamental contributions to the field.
The details I have are given below, although the costs may be negotiable! Students, those from former Eastern Europe and Third World countries are HALF price. Further (definitive!) details are available from Prof. Sitte: FAX Austria (0043 from UK) 5212 22 16 22 Mail address: Prof. Sitte, Reitherspitzstrasse 166, A-1600 Seefeld in Tirol, Austria. _______________________________________________________
The language of these workshops is English (Workshop 64, in the German language, runs from 14 to 29 October).
Workshop 63 E-Bio & E-Mat (Biomedical and Materials) Introductory sectioning workshop (without cryo or immuno) Begin 9.30 am 24 Sept. 98, finish 9 pm 27 Sept. 98 (course fee AS 7,000).
Workshop 63 F-Bio Cryoultramicrotomy, cryomethods, immuno-gold-cytochemistry Begin 9.00 am 4 Oct, finish 9 pm 8 Oct (course fee 13,000).
Workshop 63 A-Bio (combined E & F workshops, course fee AS 16,000). Workshop 63 A-Mat (combined E & F workshops, course fee AS 14,000).
Workshop B-Bio Cryofixation, freeze substitution, freeze drying, progressive lowering of temperature technique and low temperature embedding (without cryoultramicrotomy and immuno) Begin 9.00 am4 Oct, finish 9 pm 8 Oct (course fee AS 8,000).
Workshop 63 F-Mat Cryoultramicrotomy, diamond knife sectioning and preparation of hard/inhomogeneous samples. Begin 9.00 am 27 Sep, finish 9 pm 2 Oct (course fee AS 13,000).
Hi everyone, I am looking for a course on basic SEM theory, operation and hands on training. The course should not last longer than 5 days and should take place during this year. All suggestions are welcomed. Thanks! Gabriel Rojano rojanog-at-macom.com Reliability Engineering AMP- M/A-COM
I am curious as to the best way to determine the thickness of the car= bon evaporation technique being deposited on the substance to be analysed= . It would be beneficial to know the method by which a knowledge of thi= s informationcan be attained within a few Angstroms. Joshua McCaig. (mccaigjm-at-corning.com)=20 --=20 MZ=90
Hi listers, I have to fix 18.5 dpc mouse embryo in formalin or paraformaldehy for immunostaining. Does anyone out there knows the procedure for paraffin processing and embedding? We had tried several times unsuccessfully. I like to know specifically how long and in what temperature those huge embryo should be fixed (fixative penetration issue), and keep the antigenicity as well. Thanks.
******************************************************************* To see what is in front of one's nose requires a constant struggle.
George Orwell
Dorothy Zhang Harvard School of public Health Building 2, CVLAB 677 Huntington Ave, Boston, MA 02115 Phone# 617-432-6981 Fax# 617-432-2980
You may leave the water at slightly lower level even though you may not see the sections right after cutting.
Good luck !
Ming
On Tue, 21 Jul 1998, Tom Phillips wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have used LR Gold for embedding samples destined for EM } immunocytochemistry on many occassions with no trouble. In my latest } embedding, the blocks are not behaving in a typical fashion. Everytime I } go to section them, they pull the water out of the boat and prevent thin } sectioning. I can cut 0.5 um LM sections with effort and the sections look } okay. Has anybody else had this experience and, more importantly, do you } know how to get around it? TIA, tom phillips } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211 } (573)-882-4712 (voice) } (573)-882-0123 (fax) }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * #1074B Dentistry Pharmacy Building * * University Of Alberta. * * Edmonton, Alberta, Canada T6G 2N8 * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
To the lung experts, What do you do with floating lung tissue (} 2mm) that is fixed by immersion. The options are: 1)leave in a refrig overnight until they sink (swirling did not work). 2)place in a vac (15 psi) while still in the fix. 3)process the floating samples hoping they eventually sink. I am currently in the frig (opting for #1) but any hints or suggesstions would be welcome. Thanks.
We have an elderly, incomplete Leitz manual microtome about the place here. Although a possible candidate for a boat anchor, weighing perhaps 50 lb, it contains wonderful German machining and has many mysterious knobs that invite twiddling. It appears to increment (click!) after doing a slice. It is under consideration for use to section plant tissues for light microscopy at medium magnification to look at fungal pathogens at the cell level. (Not by me - I do chemical microscopy so I rarely slice.) The unit is about two feet long, resembles an infertile cross between a lathe and a bologna slicer, and the carriage is connected to a wide ribbon apparently to smooth the slicing action. It is painted gray and might plausibly date from the 1960s. It appears that the sample to be cut is intended to be clamped below, while there are clamps spanning a rather large gap (about 8 inches) which could plausibly hold a blade, although the span seems rather large (flexion). There is nothing resembling a blade holder with the microtome.
To get to the point: Can anyone supply a photocopy of a manual or pertinent parts of a manual describing the function of the parts? (fax 609-275-5239) Or perhaps be prepared to discuss how this works by telephone? Do you think it likely to be worth having a blade holder fabricated, or should we post a notice to the boat owners?
Leonard R. Corwin Fort Dodge Animal Health Cyanamid Agricultural Research Center PO Box 400 Princeton, NJ 08543-0400 609-716-2278; fax 609-275-5239 corwinl-at-pt.cyanamid.com
} 2) place in a vac (15 psi) while still in the fix.
I'd reccommend this route, specifically as follows:
Take a 2-hole stopper (or make one if you can't find one), and put the wide end of a cut-off pasteur pipette, or a length of glass tubing, into one hole. Choose a size of stopper which will seal the top of the container you're using for the fix. Use tygon tubing to connect the glass tube to house vacuum, or connect it to a similar device made with a one-hole stopper which fits onto a side-arm flask--this will prevent any oil in the line from getting to the sample. Turn on the vacuum, seal the fixing container with the two-hole stopper, place your thumb over the empty hole until the residual gas emerges from the tissue, then release the pressure. Repeat until the gas has escaped from the tissue. Note that the one-hole and two-hole stoppers can be interchanged, so if you have to hold the stopper onto the fixing container, it can be better secured if you don't have to worry about rocking the device with your thumb. Good luck. Yours, Bill Tivol
In this paper Professor Lindsay of ASU characterizes biomolecular nano-wire electron transfer using scanning probe microscopy. This characterization requires the samples are held in an environment that is free of moisture and molecular oxygen.
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; SPM Technology Leaders for Environmental / In Situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
I have cross-sectioned through gold fibers in animal tissue in the past, using a diamond knife. I did not see any problem or increased "knife marks" from these very thin ( {30 um diam.) fibers. However, I have also sectioned particles in tissue such as cobalt or titatium and one can clearly see in the photographs that there is no "knife mark" prior to hitting the particle but a HUGE one continuing beyond it, of the same thickness as the particle. By the end of sectioning a few such particle samples, that area of the knife is basically useless.
In my experience the gold doesn't dull the knife any more than animal skin, but I've never done plant tissue so I don't know how that relates.
Karen -- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
I'm in dire need of a part for my Technics Hummer III sputter coater (the plastic nut at the base of the specimen pedestal has stripped threads and cannot hold a vaccum). I can't find a current address or phone number anywhere. Can someone please reply direct??
TIA and cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
MICHAEL DELANNOY wrote: } } To the lung experts, } What do you do with floating lung tissue (} 2mm) that is fixed } by immersion. The options are: } 1)leave in a refrig overnight until they sink (swirling did not work). } 2)place in a vac (15 psi) while still in the fix. } 3)process the floating samples hoping they eventually sink.
{snip}
I never thought about it but I guess I am an expert (24 years)in TEM of lung. You did not specify whether embedding will be for LM or EM. If plastic embedding for EM, } 2mm is too thick and they should be sliced to not exceed 2mm. Other dimensions are not important as long as they will fit in an upside down Beem capsule with the pyramid end cut off.
First put them under vacuum in fix to pull out the air (the tissue will still be at the surface while in the vac) which will optimize contact of fix with tissue (I interpret your term immersion to indicate that the tissue was not first perfusion fixed or insufflated with fix).
Then they can go in the fridge or stay at room temp and should sink.
They will definitely sink during subsequent steps with osmium and acetone dehydration for TEM.
My experience does not extend to paraffin processing of lung.
Tom Christensen Pathology Boston University Medical Center
Try decreasing the water level in your boat (surface will be concave; make sure the edge is wet--if you have trouble with wetting, email back.)
Also, you can decrease the clearance angle of your knife.
Finally you might speed up the cutting stroke.
Be careful in aligning, that you don't wet the block face or the back of the knife. Don't allow the knife to stop directly at the block face; always go through a cutting stroke when checking distance, etc. The water has a way of "jumping" onto the hydrophillic block.
Good luck.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} I am curious as to the best way to determine the thickness of the carbon } evaporation technique being deposited on the substance to be analysed. } It would be beneficial to know the method by which a knowledge of this } informationcan be attained within a few Angstroms. Joshua McCaig. } (mccaigjm-at-corning.com) } -- } MZ
By using a Digital Thickness Monitor you should be able to measure carbon depositon within a few Angstroms. Since carbon is directional, it is important to locate the sensor close to the specimen. In our carbon substrate production we are able to replicate deposition within 8 to 10 Angstroms. For this it is vital that you duplicate the time, amperage, vacuum and location of the substrates. It is also important that you are consistent in your choice of carbon or graphite.
Hope this helps, John Arnott
Disclaimer: Ladd Research sells Vacuum Evaporators, Digital Thickness Monitors and substrates.
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
Mike, Please send me your web site address in order that I might see what you have to offer. Thanks again for the info on Leitz, Zeiss, and Olympus. Stephen
I have noticed this same problem with LR White when using a diamond knife. My conclusion was that after prolong cutting the knife edge becomes coated with plastic residue, and this causes the sections to bind up, causing a series of cutting problems (block wetting, sections not coming off the edge, compression). Try cutting the block with a glass knife, if this is the problem the block will cut fine, if not....back to the drawing board.
William R.McManus Supervisor Electron Microscope Facility Department of Biology Logan UT 84322-5305 billEMac-at-cc.usu.edu 435-797-1920 Fax 435-797-1575
Sometime ago, I had a Hummer III. Memory at this point could be failing, but I think the nut at the pedestal base (the one which could be loosened to raise/lower the specimen on mine) was a "bonnet" nut - either an electrical feed through bushing or a plastic tube fitting nut. Such a replacement would be common and cheap from a building supplies vendor.
Again, if memory serves.... Hummer is (was?) sold by Anatec(k?), in the Washington D.C. vicinity.
_Woody_
shAf wrote: } } I'm in dire need of a part for my Technics Hummer III sputter coater } (the plastic nut at the base of the specimen pedestal has stripped } threads and cannot hold a vaccum). I can't find a current address or } phone number anywhere. Can someone please reply direct?? } } TIA and cheerios, shAf } } {} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} } Michael Shaffer, R.A. - ICQ 210524 } Geological Science's Electron Probe Facility - University of Oregon } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
-- de Woody, WB4QXE
Work: Electron Microscopist/Microanalysist Balance: Ham radio "homebrewing", computers , shade tree mechanic "the Gravel Garage".
On the www page: Scanning Electron images and Ham Radio Homebrewing stuff. http://www.geocities.com/capecanaveral/3722 .
We run courses "in house", in your own laboratory on your own equipment, = on all aspects of electron microscopy. With the world the way it is this mea= ns we spend more than 85% of our time with SEM on - specimen preparation, instrument optimisation, x-ray analysis, maintenance and consultancy. =
Please take a look at our web site for more information, we are able to tune a course to any requirement.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
On Wed, 22 Jul 1998 12:31:20 -0400 (EDT) MICHAEL DELANNOY {delannoy-at-welchlink.welch.jhu.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To the lung experts, } What do you do with floating lung tissue (} 2mm) that is fixed } by immersion. The options are: 1)leave in a refrig overnight until } they sink (swirling did not work). } 2)place in a vac (15 psi) while still in } the fix. } 3)process the floating samples hoping } they eventually sink. } I am currently in the frig (opting for #1) but any hints or } suggesstions would be welcome. Thanks. } } Mike D }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
A friend had this problem with plant material. She kept the samples submerged under a piece of wire mesh.
Michael Shaffer asked: } I'm in dire need of a part for my Technics Hummer III sputter coater } (the plastic nut at the base of the specimen pedestal has stripped } threads and cannot hold a vaccum). I can't find a current address or } phone number anywhere. Can someone please reply direct??
The Hummer range of sputter coaters is manufactured by Anatech Ltd in Springfield, VA. The phone number is 800-752-7629 and the fax number is 703-941-8077. George Barr, the President, can certainly help you. His e-mail address is gbarr-at-anatechltd.com
Best regards, Steven Slap
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
6621 Electronic Drive Springfield, VA 22151 (800)-PLASMA-9 or 703-941-8860 FAX: 703-941-8077
Her ext. is #104.
She fixed my Technics Hummer V promptly and will assist in over the phone consultations.
Ginger
Ginger Baker EM Lab Manager OMS Secretary/Treasurer Research Dept., OCOM 1111 W. 17th St. Tulsa, OK 74107 Phone: (918) 561-8232 FAX: (918) 699-8629 http://osu.com.okstate.edu/dept/research/content/gbaker.htm lizard-at-osucom-fs02.ocom.okstate.edu
-----Original Message-----
I'm in dire need of a part for my Technics Hummer III sputter coater (the plastic nut at the base of the specimen pedestal has stripped threads and cannot hold a vaccum). I can't find a current address or phone number anywhere. Can someone please reply direct??
TIA and cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I have an old Vickers-Armstrong hardness tester. To jog your memory, it has a foot pedal load actuator and a microscope mounted on a hinged bracket, which is moved in place over the indentation for measurement. Anyway, the eye piece tube and the attached micrometer ocular has been damage by an unknown user; this is a multi-user tester. Can anyone tell me who is in the business of servicing such a beast, Canadian rep. if possible. This is a very good, solid hardness tester and I would like to keep it. They don't make them like this anymore.
Thanks in advance. David Chow National Research Council Canada David.Chow-at-NRC.CA
Microscopy/Microscopy Education also specializes in customize, on-site courses. We have over two dozen consultants here in the US who have experience on a wide variety of electron microscopes and related analytical techniques.
For more information, see our website: { {http://www.MME-Microscopy.com/education}
In regards to the answer concerning the Technics address and phone number, I hit "reply to all" instead of "reply" so email may bounce on the listserv. Sorry all.
Ginger
You need to contact Lisa Jackson at Anatech, Ltd.
6621 Electronic Drive Springfield, VA 22151 (800)-PLASMA-9 or 703-941-8860 FAX: 703-941-8077
Her ext. is #104.
She fixed my Hummer V promptly and will assist in over the phone consultations.
Ginger Baker EM Lab Manager OMS Secretary/Treasurer Research Dept., OCOM 1111 W. 17th St. Tulsa, OK 74107 Phone: (918) 561-8232 FAX: (918) 699-8629 http://osu.com.okstate.edu/dept/research/content/gbaker.htm lizard-at-osucom-fs02.ocom.okstate.edu
Ginger Baker EM Lab Manager OMS Secretary/Treasurer Research Dept., OCOM 1111 W. 17th St. Tulsa, OK 74107 Phone: (918) 561-8232 FAX: (918) 699-8629 http://osu.com.okstate.edu/dept/research/content/gbaker.htm lizard-at-osucom-fs02.ocom.okstate.edu
I need information about video printers for printing SEM images and need = to contact sales representatives from various companies. Please contact = me at 217-782-0898. Thanks. Donna Wagahoff SIU School of Medicine PO Box 19230 Springfield, Il 62794-1220
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} This prompts me to ask the obvious question. Does anyone have a foolproof } indicator to check the activity of an osmium solution, ideally without } processing tissue and viewing in the microscope? } } I 'm sure at some time we have all picked up a bottle of expensive osmium } and thought do I use it or make fresh up. This happened to me recently and I } tried soaking some osmium into a piece of cocktail stick, which appeared to } darken, but I was wrong. } } Malcolm Haswell
Malcolm, I would put a drop of corn oil (maize oil on your side of the puddle) on a slide and some of the osmium solution in a small dish (size so that the slide acts as a lid for the dish). If the osmium is good, vapors from it will blacken the oil droplet. (Any polyunsaturated oil will do.) Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Hi, All.
I am looking for a secondary detector tube for JEOL JSM 35 in a good condition. Willing to pay a fair price for it. Please reply at my email address. Many many thanks for your anticipated co-operation.
Jitu Shah
Dr.Jitu Shah H.H. Wills Physics Laboratory, University of Bristol, Royal Fort, Tyndall Avenue, Bristol BS8 1TL. UK email: jss-at-siva.bristol.ac.uk Tel: 44 117 9288719 Fax: 44 117 9255624
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Petra:
With PE/PS I have gotten good results if I first cut and then stain. I do use cryo sectioning and I cool the sample to about -100C (room temperature will not work because the Tg of PE is too low). I use folding grids to collect my sections (dry) and then I stain with RuO4 which will stain the PS. You can also stain the block first and then cut but then you are limited to sections near the surface of the specimen (the stain does not penetrate too far). Even then, you might need to re-stain your sections. Finally, I deposit carbon.
I hope this helps,
Jordi Marti ----------------------------------
Hello polymer experts,
new to the field of EM polymer blends I would like to ask you for the best way to prepare PE/PS blends for structure determination with TEM.
Of course the material has to be stained and cut, but:
- what is the best staining agent (OsO4, RuO4, or something else)? - is it better, first to stain and then to cut or the other way round? - which temperature is the best for cutting, liquid nitrogen or room temperature?
I would appreciate ever tip & trick you can give on handling this kind of material.
Tanks in advance
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Dear All, I find oil on the EDX detectors in both my SEM's, but if you think of the conditions inside the SEM, it is almost inevitable. The majority of the molecules remaining in the vacuum of an average SEM at 10 -5 torr consists of diffusion pump oil. This oil will gradually condense on all surfaces in the SEM, but on the coolest surface first. In most SEM's this is the EDX snout. One EDX manufacturer has solved the problem by warming up the snout with a small heater. The oil just goes somewhere else, but if it forms a thin film over all surfaces it is not so visible or worrying. The cleaning method recommended by the manufacturer of my EDX's is to gently run clean-grade iso-propanol over the snout, then allow to air-dry. They used to recommend Freon, but that is no longer permitted or available. Hope this helps, Mary
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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POST-DOCTORAL RESEARCH FELLOW Princeton University
A post doctoral research fellow position is available in the Princeton Materials Institute and the Princeton Center for Complex Materials at Princeton University. The appointment is for one year initially with the possibility of renewal. It involves the use of a Philips CM-200 FEG transmission electron microscope equipped with a Gatan Image Filtering System in studying carbon nanotubes, and other nanostructured materials of interest. Candidates should have a Ph.D. in the physical sciences, and with strong EM background (HREM, Electron diffraction, and EELS). Experience in image analysis and UNIX workstations, and a strong math background are highly desirable.
Applicants should send a detailed curriculum vitae, copies of 2 selected publications, and names and addresses of three referees by June 15, 1997 to:
Nan Yao Princeton Materials Institute Princeton University Bowen Hall, 70 Prospect Avenue Princeton, NJ 08540
Princeton University is an equal opportunity employer.
=============================================================== Nan Yao Princeton Materials Institute Princeton University Bowen Hall, 70 Prospect Ave. Princeton, New Jersey 08540-5211
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} To All } I have been doing TEM since 1970, and have always used NaOH pellets } (CP grade - which do contain about 0.5 - 1% Na2CO3). I make up a 10N } solution in small aliquots (~20 ml), and dump it when I see crystals } of sodium silicate (NaOH dissolves glass). I use this 10N solution } to add to the powdered Pb citrate in dH2O. I make up Pb citrate in } 100 ml quantities and dump it when it turns cloudy. Usually it keeps } for months, unless someone forgets to tighten the cap, which is not } uncommon in a central service lab. What I have learned in nearly 30 } years of TEM, and many years in other microscopy techniques is that } there are very few empirical rules. I have been in very fussy labs } that used aged Pb citrate in bottles that were so coated with } precipitate that they looked out of a sunken pirate ship, and have } been in other labs that make up Pb citrate fresh, and filter it. } Whatever works. } Bruce Cutler, Microscopy Lab, Univ. Kansas, Lawrence KS
I store our freshly made-up concentrated Reynolds solution in 3ml aliquots in Eppendorff tubes at 4C. This way there is minimal exposure to CO2 and no danger of someone leaving the container open. A tube of stain is only used once, anything left over in the tube being discarded. The stain keeps well for months in these tubes.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377 Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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If I am off-base with this response, I apologize. I have used nothing but carbon tape ever since it became readily available through regular EM suppliers but I always dreaded having to clean it off my sample holders because it was so sticky and tore so easily. Being a simple minded person, I just took a 3/8" wooden dowel, sharpened one end(like a chisel) and used it to roll the tape up into a wad that I could remove with my fingers. All the adhesive seems to comes off with the tape, but if any should be left behind it should easily come off with a mild solvent. (I always polish my holders, so I'm not sure about residual adhesive.) I also cut a piece of latex tubing to fit on the end of the rod to cushion my hand. This works for the carbon tape, I'm not sure about the other types of adhesive tapes. For what it's worth.
Bill -- ============================================================= Bill Chissoe III Electron Microscopist,University of Oklahoma E-mail: wchiss-at-ou.edu Ph. (405)325-4391 ============================================================= Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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You wrote: " I am looking for the fax number or email address of Scanning Microscopy Int. (Chicago) - can anybody help?
Thanks in advance Fiona Graham Electron Microscope Unit University of Natal, Dalbridge, 4041, South Africa tel: +27 31 260 2174 fax: +27 31 261 6550 email: GRAHAM-at-ph.und.ac.za"
I have used 73211.647-at-compuserve.com within the last couple of months and it was OK.
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 617-275-4695 FAX: 617-275-4695 Alternate FAX: 617-271-0252
email dmrelion-at-world.std.com
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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JOINT SPRING MEETING
MISSOURI-ILLINOIS-KANSAS MICROBEAM ANALYSIS SOCIETY & CENTRAL STATES MICROSCOPY SOCIETY
Presentations will be in the Arthur Mag Conference Center (Mag Center) of Midwest Research Institute, Kansas City, MO
APRIL 25, 1997 ====================================================== PROGRAM
8:30-9:00 Registration, Vendor Display Setup in the Mag Center.
9:00-9:10 Welcome by Dr. William P. Duncan, Director, Midwest Research Institute.
9:10-9:25 Dr. Peter Moroz, Jr., G.S. Technologies, Kansas City, MO "Microstructure of Metal"
9:30-9:45 Harold McCormick, C-K Engineering Inc., St. Louis, MO "Wearever of Metals"
9:50-10:05 Garry Crabtree, TechSpec Inc., Raytown, MO "Material Response to Deep Cryogenic Tempering"
10:10-10:30 Morning Break-Refreshments from BAR ROMA (cash cart).
10:30-10:45 Dr. Ody Maningat, Midwest Grain Products, Inc., Atchison, KS "Wheat Starch and Wheat Gluten Research"
10:50-11:05 Dr. Diane Durham, KU Medical Center, Kansas City, KS "Regeneration of Sensory Hair Cells in the Avian Cochlea-SEM Analysis"
11:10-11:25 Dr. Peter Smith, KU Medical Center, Kansas City, KS "Light and Electron Microscopic Investigation of Nervous System Plasticity"
11:30-11:45 Dr. Amit Mukherjee, KU Medical Center, Kansas City, KS "Electron Microscopic Analysis of the Polymerization of FtsZ, an Essential Cell Division Protein of E-Coli"
12:00-1:15 BUFFET LUNCH Generously provided by HITACHI, catered by Nance's Deli and Catering. Buffet includes vegetable manicotti, garden salad, garlic bread, soft drinks, etc.
1:15-1:35 Business Meetings
1:35-1:55 Paul Benson, The Nelson-Atkins Museum of Art, Kansas City, MO "Scientific Technique as Applied to Art Objects"
2:00-2:15 Marv Hart, Century Lubricants, Kansas City, KS "Lubrication and Grease Technology"
2:20-2:35 Garth Kristoff, Allied Signal, Kansas City, MO "Overview of the Technical Transfer and Solvent Substitute for Electronic Cleaning"
2:40-3:00 Afternoon Break-Refreshments from BAR ROMA (cash cart)
3:00-3:30 Michael Saba, Digital Instruments, Eden Prairie, Minnesota. "Applications in Scanning Probe Microscopy"
3:35-3:50 John Wilson, Mo. Regional Criminalistic Lab, Kansas City, MO "Capabilities of Crime Scene Investigation"
****************************************************** Complimentary lunch will be provided for all attendees by HITACHI. However, we need to know how many will be attending the luncheon, prior to the meeting. Please phone or email one of the following:
Larry Irwin 913-268-9009 Dan Kremser 314-935-5605 dkremser-at-levee.wustl.edu
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Peter Jordan wrote: } } Hi: } I have received two E-mail letters from Philip Koeck over the past week } or so through this forum. Both letters I can not read or delete. } Whenever I try I get an error message saying that the program has } performed an illigal action and turns netscape off. Anybody having the } same problem or any solutions? Thank you, Peter Jordan .......................... I also use Netscape 3.0 and have the same problem. I don't have a solution, but I do have a way to get rid of the "poison message":
(1) read, and delete or transfer all the rest of the mail in the Inbox, but don't try to read or delete the "problem" message. (This is tricky, and you may have to "error-out" and reload Netscape a couple times.) (2) When the Inbox is empty except for this message, exit from Netscape. (3) Go to the NETSCAPE\NAVIGATOR\MAIL subdirectory, and locate the file "Inbox". Delete it (or rename it if you want to be cautious). (4) The next time you load Netscape, it will recreate an empty Inbox file.
This is ugly, but it's the only way I've found to get rid of the darn thing!
I hope somebody who doesn't have this problem will help us out by sending a message to the originator to find out what he is doing! I can't open his messages to get a return address. -- Fred Schamber ....................... mailto:fhscham-at-SGI.NET Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Hi, Everyone,
My background is in EM of biological, medical and food samples. I now have the opportunity to expand my skills and work on a set of materials samples.
My question is:
What are the standard methods for assessing surface degradation of materials samples (especially plastic polymers)? Are there macroscopic as well as microscopic methods which can give statistically "good" descriptions of the extend and type of degradation of such surfaces?
Thanks for any help (methods, references, review papers, etc.) you can provide. Please contact me offline.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia,Canada B4N 1J5
tel: (902) 679-5566 fax:(902) 679-2311 e-mail: allanwojtasp-at-em.agr.ca Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Keith: I can't help you with sonicating water, but there is only one thing to do with flat beer, bin it. But then you people in the West Country have no idea what constitutes good beer. Come to Cambridge for a pint of Greene King Abbot or, better still Adnams. Both will knock your socks off
PatrickOn Wed, 16 Apr 1997, Keith Ryan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Ian } } I really appreciate the tip about sonicating water to degas it, I would not } have believed it. Now, do you have any suggestions for flat beer? } } Regards - Keith Ryan } Plymouth Marine Lab., UK } } }
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Would freshly distilled water be equivalent to degassing via sonication ?
Leo
On Wed, 16 Apr 1997, Ian Montgomery wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } Good morning Reynolds users } } } } In nearly 30 years of preparing lead citrate according to } } Reynolds I have experienced no problems in using a stock } } sodium hydroxide solution (1M) which was made up from pellets. } } However, this solution must be freshly made up. Is it because I } } always use Reynolds in its concentrated form that I do not } } experience any problems? } } } } Rob } } } Like Rob I've been making Reynold's lead citrate since I was a boy. } I use NaOH pellets but degas the distilled water by sonicating it for a few } minutes before making the stain. } } Ian. } } } Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
Peter Jordan wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi: } I have received two E-mail letters from Philip Koeck over the past week } or so through this forum. Both letters I can not read or delete. } Whenever I try I get an error message saying that the program has } performed an illigal action and turns netscape off. Anybody having the } same problem or any solutions? Thank you, Peter Jordan
Yes I am. It's very frustrating. I'd appreciate any help.
Thanks.
Eric Eric H. Metzler 1241 Kildale Sq. N. Columbus Ohio 43229-1306 USA
Phone: 614 888 3642 E-mail: spruance-at-infinet.com Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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} For Sale: JEOL JSM T-200 scanning electron microscope. Purchased in } 1984; used about 50 hours, total time. Includes chiller and a small } sputter coater. $1500 OBO. Contact Dr. Jon Martin at 912/752-4060. } Located at Mercer University School of Medicine, Macon Georgia.
E-mail contact: HORST_MN-at-Mercer.EDU ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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If you have "copies of copies" which work ok, and you have suitable blank media, there is no reason you cannot copy these again to generate archive disks. If you have a "verify" command/switch - use it. ...Or am I missing something??
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We have a NORAN (Tracor Northern) 5400 Series II EDS system (Model Number TN-5402/BBAA) acquired in 1987. It does not have a hard drive and operates off of two floppy drives. It requires 5.25" double sided, high density floppy disks, the originals of which are no longer readable. They have not been in use other than to be the source of copies made periodically but apparently they have simply deteriorated with time. We have copies of copies that are working pretty well but I'm becoming concerned that they may crash/die and we'll have no good source from which to make new copies. NORAN does not have replacement masters of this vintage so I'm hoping to find someone who has original master disks in good working order who would be willing to loan them to be copied.
We need two disks of the set from the November, 1987 Release 1C. What will work with our system is very specific and the original label on the master disks reads:
SERIES II SOFTWARE RELEASE 1C NOV., 1987, TRACOR NORTHERN, INC.
Of the disks in this release, we need:
SQ/SSQ/PRZ/ZAF Master NOT BOOTABLE
and
MSCAN2 Master
Thanks in advance for any help or suggestions you may have.
___________________________________________________________________________ Barbara Reine, Botany Dept. Box 351330 Univ. of Washington, Seattle, WA 98195-1330 e-mail: reine-at-u.washington.edu; ph: (206) 543-1955 ____________________________________________________________________________ Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
id NAA42426 (8.8.6/50); Thu, 23 Jul 1998 13:02:14 -0500 Message-Id: {199807231802.NAA42426-at-mail1.doit.wisc.edu} X-Sender: steve-at-facstaff.wisc.edu (Unverified) X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
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To all who responded to my plea for help: Thank you very much. Sandwiching the "bad" files between two "good" files allows you to move them into the trash bin and then to delete them. For all computer greenhorns like me this is done by clicking on the good file, then holding down the shift key clicking on the other good file. All files between the clicked files will be marked and then can be moved. It was realy nice to get all these respones. Thanx again, Peter Jordan Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Hello all,
We work as consultants for a computer imaging hardware and software manufacturer's rep, and have had a rather unusual request. Perhaps someone, either end users or vendors can assist.
We are looking for the equivalent of a MacBeth Color Chart, typically used in video, but this needs to be very small, translucent, and mounted on a standard histo glass slide. Ideally, 1/32" square (1 mm x 1 mm OK) and it would contain at least the primary colors, incl. black and white. I suspect there may be a problem with ?translucent black and white. I think episcopic illumination would also suffice for the chart, however so it could be opaque. At this time, I do not have more detail on their exact application.
We have checked with Munsell and MacBeth to no avail, so we would appreciate any assistance, and even a referral as it's likely this is a custom application. You can e-mail me directly if you like.
Thanks in advance!!
Daryl Martin dmartin-at-ic.net
(313) 213-8444
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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} What are the standard methods for assessing surface degradation of } materials samples (especially plastic polymers)? Are there macroscopic } as well as microscopic methods which can give statistically "good" } descriptions of the extend and type of degradation of such surfaces? } Dear Paula, I recently ran across a good review "Electron Microscopy in Polymer Science" by G.H. Michler, Applied Spectroscopy Reviews, vol 28(4), pp 327-384 (1993). This paper discusses surface characteriza- tion and many other topics. Additionally, I suggest STM or AFM as possibilities, but I don't have any referrences for them. Perhaps comparing the specular vs diffuse reflectivities of polymer surfaces before and after damage would be informative for degradation features ~ 1 micrometer in size. Good luck. Yours, Bill Tivol Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Meeting Announcement -
MAMAS (Mid-Atlantic Microbeam Analysis Society) and the Surface and Microanalysis Science Division, NIST Meetin at the National Institute of Standards and Technology, Gaithersburg, MD on Thursday, May 15, 1997, 10:30 am- 3:00 PM Lecture Room D, Administration Bldg.
10:30am Coffee and Doughnuts
10:45am Prof. David R. Veblen, Dept. of Earth and Planetary Science, Johns Hopkins University "Transmission Electron Microscopy of Minerals"
12 noon Lunch
1:15pm Dr. Michael Kersker, TEM/STM Project Manager, JEOL USA "200 KV FEG: Refried Beans with a Fiery New Salsa"
For more information, contact Ryna Marinenko (301)975-3901, FAX(301)417-1321, email:ryna.marinenko-at-nist.gov
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There is another option for scanning film negatives. A Leaf MicroLumina camera can be coupled with a high frequency light box and a copy stand to provide a simple and highly flexible solution to the problem of scanning film and transparencies.
The camera can be racked up and down on the stand to provide a field of view from 1" X 1.5" up to 9" X 12". The camera has a resolution of 2700 X 3380 pixels and can capture 12 bit grayscale and 36 bit color.
This configuration is currently offered by our company to the electron microscopy community under the name TEMSCAN.
If anyone is interested in this product, please contact us by phone: 516-773-4305 or e-mail: sales-at-electroimage.com. Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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PROPOSED IMMUNOCYTOCHEMISTRY NEWSGROUP
This is THE FINAL URGENT REQUEST for your comments, suggestions, etc about the 3RD RFD for my PROPOSED NEW NEWSGROUP SPECIALIZING IN IMMUNOCYTOCHEMISTRY/IMMUNOHISTOCHEMISTRY/OTHER RELATED AFFININITY METHODS called "sci.bio.immunocytochem" now posted in "news.groups"
The latter contains material of a rather varied nature (!) but it is necessary to use this unmoderated group to hold the discussions about all the different proposed new groups.
So, if you are connected to Usenet, and you are keen to see A NEW IMMUNOCYTOCHEMISTRY NEWSGROUP, then please go to "news.groups", ignore all the rubbish, look for articles posted on 20.4.97 or 24.4.97 (or later) and you should find one of my postings "3RD RFD: sci.bio.immunocytochem". Select "follow-up article" (or equivalent) from your newsreader menu, and post your message so that it appears under the 3RD RFD.
If you don't have access to Usenet, you can read the proposal at the Royal Microscope Society web site {http://www.rms.org.uk} , or at the Introduction to Immunocytochemistry (Center for Cell Imaging Department of Cell Biology Yale University School of Medicine) web site {http://info.med.yale.edu/cellimg/CCIimmuno.html}
MANY THANKS to all of you who have already posted your responses to my RFDs! BUT WE STILL NEED MORE DISCUSSION in news.groups please! SOON I shall be posting my "CALL FOR VOTES" to the people in charge of Usenet newsgroups. When you see "CFV: sci.bio.immunocytochem", you will be able to e-mail your vote to the vote-taker.
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Message on behalf of Richard Easingwood; } } We are looking at the various options for large-format (4 x 5 inch) } negative scanners and wonder whether we could get some feedback from actual } users. } I have heard that the Leaf 45 scanner (which I understand was the Rolls } Royce/Cadillac of scanners) is discontinued and no longer available - can } anyone confirm this? } } Does anyone have experiences (good or bad) with the Polaroid SprintScan 45? } } We want to use the scanner for getting high quality resolution scans from } our sheet film TEM micrographs. } } Thanks in advance for any feedback. } } Regards, } } Richard } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences } University of Otago } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } e-mail: richard.easingwood-at-stonebow.otago.ac.nz } } }
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Good morning Reynolds users
In nearly 30 years of preparing lead citrate according to Reynolds I have experienced no problems in using a stock sodium hydroxide solution (1M) which was made up from pellets. However, this solution must be freshly made up. Is it because I always use Reynolds in its concentrated form that I do not experience any problems?
Rob
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377 Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Thanks to the several people who responded to my question.
The following was received directly from Bart Cannon and has some useful detail. I thought others might be interested.
} Arthur Wehnelt (1871-1944) was the German physicist who invented the } "Hot Cathode" electron tube which employed what was called a "grid" to } direct a stream of electrons. It corresponds fundamentally to the } electron gun used in oscilloscopes, TVs, and electron microscopes. Grid } and wehnelt have been interchangable terms to some degree.
-- Fred Schamber ....................... mailto:fhscham-at-SGI.NET Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Dear Mark,
We buy our plastic storage pages from the University bookstore and also from local photography stores. They are archival quality and come in a variety of sizes. They are made by Print File Archival Preservers, PO Box 607638, Orlando Fl, Ph:407-886-3100; Fax 407-886-0008. The type I use for 4x5 polaroid prints is product #45-8P (which are a heavier weight than the negative preservers).
Hope this helps.
Rosemarie Rosell Assistant Research Scientist Dept. of Plant Sciences University of Arizona Tucson,AZ 85721 Ph: 520-621-1230 Fax: 520-621-8839
On Wed, 30 Apr 1997, Mark Blackford wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All, } } I am trying to find a source of plastic pages for three-ringed binders } which have pockets large enough to hold polaroid prints (these are 131mm x } 106mm). Ideally each page should have 4 pocket and be able to fit two } prints back-to-back so images are visible from both sides of the page. } } I would really appreciate it if someone could give me an } address/phone/fax/email of a supplier of these items (if, indeed, these } things are made). Cheers, } } Mark Blackford } TEM Group } Materials Division, ANSTO } PMB 1, } Menai, N.S.W. } Australia } 2234 } Phone 61 2 9717 3027 } Fax 61 2 9543 7179 } } Disclaimer: } The views expressed in this E-mail message do not necessarily represent the } official views of ANSTO from which this message was conveyed. } } } Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Hi there,
I'm looking for carbon EM calibration grid from SPI model 411CG-AB but couldn't find any information about SPI.Does anyone knows they phone # or www address? Thanks.
**************************** * Maoxu Qian, Ph.D. * * Dept of MSE, box 352120 * * University of Washington * * mxq-at-u.washington.edu * * (206)543-1514(phone) * * (206)543-3100(fax) * ****************************
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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At 04:46 PM 4/29/97 +1200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, this is true. However, you may be able to locate a refurbished Leaf 45 Scanner by contacting a local digital photography distributor.
} } } } Does anyone have experiences (good or bad) with the Polaroid SprintScan 45? } } } } We want to use the scanner for getting high quality resolution scans from } } our sheet film TEM micrographs. } } } } Thanks in advance f snip, snip
A new option which I have not seen discussed on this forum is a drum scanner. The company ScanView which has made high-quality drum scanners for the pre-press market has now begun to market cheaper drum scanners at a relatively low price. We recently purchased the Scanview 3000 (3000 dpi, ~$15,000) which has an optical density range of 3.6, color resolution of 3 x 12 bit, 4096 levels, and a scanning area of 8.5" x 11.5". Some advantages of this scanner over the Leaf 45 are that the 3000 dpi resolution is available over the entire drum and the scan speed is much faster at the same resolution since it is a single pass scan. So, enlargements of an area of the negative which is off center are easy to do. You can also mount up to 6 negatives at once and set up batch jobs which will free up some of your time. We have been very pleased with the results so far. I believe it will deliver all possible information from the negative since the resolution of negatives are not much better than 3000 dpi in most circumstances. ScanView also makes two cheaper drum scanners the Scanmate Plus II (2600 dpi, o.d.=3.6) and the Scanmate Magic (2000 dpi, o.d.=3.0). I suggest that you contact your local digital photography dealer for a demonstration.
I have no financial interest in ScanView. I am just a satisfied user.
******************************************************* David F. Teter Los Alamos National Laboratory Materials Science and Technology: Metallurgy (MST-6) Mail Stop: G755 Los Alamos, NM 87545 ph: (505)665-0160 fax: (505) 665-0657 e-mail: teter-at-lanl.gov *******************************************************
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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NYMA Inc., an aerospace engineering company serving NASA at Lewis Research Center, is seeking to fill the following two challenging positions;
SCANNING ELECTRON MICROSCOPIST
Required to operate and maintain three scanning electron microscopes, and provide materials characterization support. The individual must also be able to assist and instruct research staff in the principles and operation of scanning electron microscopy.
Requirements: B.S. degree with 3 years experience or 7+ years of extensive hands on experience in electron microscopy. Must have a thorough working knowledge of x-ray energy/wavelength dispersive spectroscopy, preparation equipment, and vacuum systems. A working knowledge of x-ray diffraction and electronics is a plus. Excellent communication and interpersonal skills are required.
TRANSMISSION ELECTRON MICROSCOPIST
Extensive experience in operating, and maintaining a 200 KeV transmission electron microscope in support of material characterization of advanced high temperature materials. The successful candidate will work with materials researchers to understand materials' properties.
Requirements: Ph.D. in material science with 3+ years extensive experience operating a transmission electron microscope using SAED, CBED, XEDS, EELS, and PEELS to analyze a wide range of materials. The individual we seek must have extensive experience in sample preparation using electro-polishing, ion-milling, PIPS, and PIMS. A working knowledge of a dedicated STEM and x-ray diffraction is a plus. Excellent communication and interpersonal skills are required.
Qualified candidates should submit a resume to : Todd Leonhardt, NYMA, Inc., Mail Stop 105-1, 2001 Aerospace Parkway, Brook Park, OH 44142 or by E-mail to todd.a.leonhardt-at-lerc.nasa.gov
Posted Date: April 28,1997 Closing Date: Open until filled
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Fellow microscopists I am doing some TEM of cultured lymphocytes. All membranes seem to be absent. There are halos where membranes should be but no membranes. So far I have tried 2.5 glut in 0.1M cacodylate 2.5 glut in 0.2M cacodylate (caused shrinkage) 4 paraformaldehyde 1 glut in 0.1M cacodylate.
Post fixing in Osmium dehydrating in ethanol and embedding in Spurr
I am going to try making up the fix in the culture medium next.
Has anyone any thoughts of anything else to try. I would be particularly interested in the use of Ca+ Mg+ and sucrose.
Many thanks
Chris Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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G'day Colleagues...
Just a quick note to let you all know that that Microscopy Listserver Archives are now on-line. I have made accessible all Email postings covering the period Oct.1993 through Mar.1997 and will update the archive monthly.
The index is not directly searchable, however, it is chronologically sorted by Month and Year.
If you download a given month's postings then you can search the downloaded WWW page for any keyword/phrase that you wish by using the native search/find option of your WWW Browser. (In NetScape this is located in the Edit Pull Down Menu and is called FIND).
You may access the archive at the MSA WWW site.
http://www.msa.microscopy.com
just follow the links to the Reference/Educational Activities Page.
I'll get around to putting together a completely searchable index sometime in the forseeable future, but for now this will go a reasonable way to letting everyone find "old messages and postings" and all the other miscellaneous requests I receive for information.
Cheers...
Nestor Your Friendly Neighborhood SysOp.
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Aloha, y'all (I've just been in Texas)
Below is a request for instructions and/or parts for a cryostat from some colleagues.
Thanks in advance for your help!
Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* **************************************************************************** Message:
We have recently obtained a surplus cryostat, a Milles Scientific Microtome Model 4553. The unit chills well and the microtome is mechanically in good shape. Unfortunately, it does not have instructions, an antirolling plate or specimen stubs. Does anyone know a source for these items? We have been unable to locate a phone number or address for Milles Scientific. Perhaps they have gone out of business or merged with another company. Perhaps someone has surplus parts for this model we could acquire. Your assistance would be appreciated.
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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I am working with distarch phosphates (carbohydrate chains covalently linked together by Phosphorous), created with POCl3. I was hoping that someone can tell me whether it would be possible to use some type of probe (maybe with fluorescence?) to be able to detect the phosphorous based crosslinks. I have used SEM with EDS and this technique is not sensitive enough to detect the low level of phosphorous in the starch.
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Negafile has polaroid 4x5 pages 4 per sheet. I'm not sure if they have gone to one size now or if they still have the two 4x5 sizes. You used to have to specify an "L" in the part number when ordering, but the number on the sheet itself, 4504, was the same for both. Check with them. I do not have their phone number, but their address is P.O. Box 78, Furlong, PA 18925, USA
- -Scott Walck
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UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Dear all
I may be dragging the thread away from contamination, but I thought that most electron microscope users avoided silicon based oils in diff pumps etc
because they are extremely difficult to remove from the interior of an electron microscope and any contamination would normally have electrical insulating properties (catastrophic in an e.m.). Perhaps I am wrong but I would welcome any comments. After all this is one of the reasons why Santovac oils and their relatives became so popular (despite their costs).
If I am labouring under a mis-apprehension then I apologise.
Malcolm Haswell University of Sunderland UK
All of the silicon based DP fluids I am aware of (which may not be all that are on the market) will break down under an electron beam and deposit a layer similar to glass on the nearest cool surface in the column. I don't know of any EM manufacturers that would recommend their use because they are almost impossible to remove if they do get in the column. We don't even use silicon based fluids in our vacuum evaporator.
K. A. Brackett, Ph.D. TN & Assoc./USEPA
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Petra;
I have a good reference that you might want to check out:
"Ruthenium Tetraoxide Staining of Polymers for Electron Microscopy", by J.S. Trent, et al, Macromolecules Vol. 16, #4, pp. 589- 1983.
This article is VERY informative.
Regards,
Bob *********************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com ***********************************
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Hello polymer experts,
new to the field of EM polymer blends I would like to ask you for the best way to prepare PE/PS blends for structure determination with TEM.
Of course the material has to be stained and cut, but:
- what is the best staining agent (OsO4, RuO4, or something else)? - is it better, first to stain and then to cut or the other way round? - which temperature is the best for cutting, liquid nitrogen or room temperature?
I would appreciate ever tip & trick you can give on handling this kind of material.
Tanks in advance
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
id NAA41274 (8.8.6/50); Thu, 23 Jul 1998 13:02:43 -0500 Message-Id: {199807231802.NAA41274-at-mail1.doit.wisc.edu} X-Sender: steve-at-facstaff.wisc.edu (Unverified) X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
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Ultrasonication has been a standard technique for degassing fluids in our labs for many years. (Before that we boiled where possible or pulled a gentle vacuum on a closed container. Main purpose was to degas solutions to be passed through automatic light blockage partcle counters. (Air bubbles count very nicely as particles.) Nice thing about sonication is that you can safely sonicate oils and other fluids that you wouldn't want to boil or pull into a vacuum system.
Bob Holthausen Pall Corporation
} } On Wed, 16 Apr 1997, Ian Montgomery wrote: } } } Like Rob I've been making Reynold's lead citrate since I was a boy. } } I use NaOH pellets but degas the distilled water by sonicating it for a few } } minutes before making the stain. } } } } Ian. } } Leo, If my memory serves I got sonication from the Technical Hints and Tips in the Proceedings of the RMS. I cover my options by sonicating the water whether fresh or hours, days old. Ian.
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
I would like to thank everyone who responded to my request of information about digital cameras. I am sorry that I did not reply to you individually. No more information, please!
Thank you very much for your kind help.
Hong You Dept. of Cell Biology University of Cincinnati
You may want to check with Bruce D. Newell, Scientific Imaging Systems, Eastman Kodak Co., 343 State Street, Rochester, NY, 14652. In his talk at Microscopy and Microanalysis '98, in Atlanta (9 AM, Tuesday, 14 July 1998), he showed a slide of a tiny Macbeth chart meant to be viewed under a light microscope. I don't think it is being produced commercial yet (I asked at the Kodak Exhibitor's booth).
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
On Thu, 23 Jul 1998 steve-at-facstaff.wisc.edu wrote:
} Hello all, } } We work as consultants for a computer imaging hardware and software } manufacturer's rep, and have had a rather unusual request. Perhaps someone, } either end users or vendors can assist. } } We are looking for the equivalent of a MacBeth Color Chart, typically used } in video, but this needs to be very small, translucent, and mounted on a } standard histo glass slide. Ideally, 1/32" square (1 mm x 1 mm OK) and it } would contain at least the primary colors, incl. black and white. I suspect } there may be a problem with ?translucent black and white. I think episcopic } illumination would also suffice for the chart, however so it could be } opaque. At this time, I do not have more detail on their exact application. } } We have checked with Munsell and MacBeth to no avail, so we would appreciate } any assistance, and even a referral as it's likely this is a custom } application. You can e-mail me directly if you like. } } Thanks in advance!! } } Daryl Martin } dmartin-at-ic.net } } (313) 213-8444 } } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu }
You may want to check with Bruce D. Newell, Scientific Imaging Systems, Eastman Kodak Co., 343 State Street, Rochester, NY, 14652. In his talk at Microscopy and Microanalysis '98, in Atlanta (9 AM, Tuesday, 14 July 1998), he showed a slide of a tiny Macbeth chart meant to be viewed under a light microscope. I don't think it is being produced commercial yet (I asked at the Kodak Exhibitor's booth).
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
} Hello all, } } We work as consultants for a computer imaging hardware and software } manufacturer's rep, and have had a rather unusual request. Perhaps someone, } either end users or vendors can assist. } } We are looking for the equivalent of a MacBeth Color Chart, typically used } in video, but this needs to be very small, translucent, and mounted on a } standard histo glass slide. Ideally, 1/32" square (1 mm x 1 mm OK) and it } would contain at least the primary colors, incl. black and white. I suspect } there may be a problem with ?translucent black and white. I think episcopic } illumination would also suffice for the chart, however so it could be } opaque. At this time, I do not have more detail on their exact application. } } We have checked with Munsell and MacBeth to no avail, so we would appreciate } any assistance, and even a referral as it's likely this is a custom } application. You can e-mail me directly if you like. } } Thanks in advance!! } } Daryl Martin } dmartin-at-ic.net } } (313) 213-8444
We are working with Bruce to provide the color chart commercially and look to have a product this fall. We will put a posting at our website when it is available:
{ {http://www.MME-Microscopy.com/education}
Please check in September. ... Thanks for the interest.
We are working with Bruce to provide the color chart commercially and look to have a product this fall. We will put a posting at our website when it is available:
{ {http://www.MME-Microscopy.com/education}
Please check in September. ... Thanks for the interest.
-----Original Message----- From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] Sent: Thursday, July 23, 1998 7:52 PM To: MICHAEL DELANNOY Cc: microscopy-at-Sparc5.Microscopy.Com Subject: Re: Floatin lung tissue
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On Wed, 22 Jul 1998 12:31:20 -0400 (EDT) MICHAEL DELANNOY {delannoy-at-welchlink.welch.jhu.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To the lung experts, } What do you do with floating lung tissue (} 2mm) that is fixed } by immersion. The options are: 1)leave in a refrig overnight until } they sink (swirling did not work). } 2)place in a vac (15 psi) while still in } the fix. } 3)process the floating samples hoping } they eventually sink. } I am currently in the frig (opting for #1) but any hints or } suggesstions would be welcome. Thanks. } } Mike D }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
A friend had this problem with plant material. She kept
the samples submerged under a piece of wire mesh.
Dave
Mike,
Place gauze over the top of the fixative and tissue, this prevents the tissue from rising to the surface and drying out.
-----Original Message----- From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] {mailto:[mailto:David.Patton-at-uwe.ac.uk]} Sent: Thursday, July 23, 1998 7:52 PM To: MICHAEL DELANNOY Cc: microscopy-at-Sparc5.Microscopy.Com {mailto:microscopy-at-Sparc5.Microscopy.Com} Subject: Re: Floatin lung tissue
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On Wed, 22 Jul 1998 12:31:20 -0400 (EDT) MICHAEL DELANNOY {delannoy-at-welchlink.welch.jhu.edu {mailto:delannoy-at-welchlink.welch.jhu.edu} } wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} } -----------------------------------------------------------------------. } } } To the lung experts, } What do you do with floating lung tissue (} 2mm) that is fixed } by immersion. The options are: 1)leave in a refrig overnight until } they sink (swirling did not work). } 2)place in a vac (15 psi) while still in } the fix. } 3)process the floating samples hoping } they eventually sink. } I am currently in the frig (opting for #1) but any hints or } suggesstions would be welcome. Thanks. } } Mike D }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk {mailto:David.Patton-at-uwe.ac.uk} "University of the West of England"
A friend had this problem with plant material. She kept
the samples submerged under a piece of wire mesh.
Dave
Mike,
Place gauze over the top of the fixative and tissue, this prevents the tissue from rising to the surface and drying out.
-----Original Message----- From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] {mailto:[mailto:David.Patton-at-uwe.ac.uk]} Sent: Thursday, July 23, 1998 7:52 PM To: MICHAEL DELANNOY Cc: microscopy-at-Sparc5.Microscopy.Com {mailto:microscopy-at-Sparc5.Microscopy.Com} Subject: Re: Floatin lung tissue
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On Wed, 22 Jul 1998 12:31:20 -0400 (EDT) MICHAEL DELANNOY {delannoy-at-welchlink.welch.jhu.edu {mailto:delannoy-at-welchlink.welch.jhu.edu} } wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} } -----------------------------------------------------------------------. } } } To the lung experts, } What do you do with floating lung tissue (} 2mm) that is fixed } by immersion. The options are: 1)leave in a refrig overnight until } they sink (swirling did not work). } 2)place in a vac (15 psi) while still in } the fix. } 3)process the floating samples hoping } they eventually sink. } I am currently in the frig (opting for #1) but any hints or } suggesstions would be welcome. Thanks. } } Mike D }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk {mailto:David.Patton-at-uwe.ac.uk} "University of the West of England"
A friend had this problem with plant material. She kept
the samples submerged under a piece of wire mesh.
Dave
Mike,
Place gauze over the top of the fixative and tissue, this prevents the tissue from rising to the surface and drying out.
I have got some requests on a summary of the answers concerning my recently posted question. I wish to express my thanks for the three answers I received: --- } Try the very recent reference in } Journal of Histochemistry and Cytochemistry vol.46, No.3 (March 1998) } } "TUNEL Apoptotic Cell Detection in Tissue Sections: Critical Evaluation and } Improvement" by Labat-Moleur,Francoise et. al. in Grenoble, France. } } Raymond Koelling } Immunex Corp. } Dept. of Molecular Immunology } 51 University Street } Seattle, WA 98101 --- } Check out Boehringer Mannheim's website http://biochem.beohringer.com } It has a complete Guide to Cell Proliferation and Apoptosis Methods. } } Ronnie Houston } Cytochemistry & Molecular Pathology } Texas Scottish Rite Hospital for Children } 2222 Welborn Street } Dallas, TX 75219 --- } This is a protocol that I used a few years ago to check for } apoptosis. My samples were embedded in paraplast plus. } } TUNEL assay for paraffin embedded samples } } 1) Deparaffinze & rehydrate to buffer } 2) Proteinase K digestion 15 min } 3) Distilled water rinses 3X 5 min } 4) Air dry } 5) Reaction Mixture (8-10 ul under coverslip), humid, 37C 60 min } 6) Distilled water to remover the coverslip } 7) Distilled water wash 20 min } 8) 0.1M Sodium pyrophophate (make fresh) 10 min } 9) 0.1M Periodic acid (make fresh) 10 min } 10) Distilled water 5 min } 11) 2% BSA 20 min } 12) Horse radish peroxidase-avidin 30-45 min } 13) Tris buffered saline 3X 5 min } 14) DAB or AEC } } Counterstain with Methyl Green } } The reaction mix came from a Boehringer Mannheim Terminal transferase kit. } The Proteinase K was 10ug Pro.K/ml 50mM TRIS, pH 8.0, use 100 ul per section. } I used either the HRPO-avidin at 1:1000 with 2% BSA + 0.5M NaCl or a ready } to use solution of streptavidin-HRP from BioGenex. } The AEC came from a BioGenex substrate kit. } Reaction Mixture (use 10ul per section): } use a glass coverslip to cover the slide & incubate at a humid 37 degrees C } 100ul 5x TdT buffer (labelling buffer) } 100ul 1:4 diluted 25 mM CoCl2 } 10ul 1mM biotin-16-dUTP } 7.5ul TdT } 282.5ul distilled water } } This used to work very well for me, if you have any questions please feel } free to contact me. } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } phone: 510-642-2085 } fax: 510-643-6207
Thank you very much again.
-- Mit freundlichen Gruessen Yours sincerely ************************************************************** * PD Dr. T. J. Filler | specialist in anatomy * * Westfalian Wilhelms-Univ.| phone: *49 (0) 251 83 552 26 * * Institute of Anatomy | fax: *49 (0) 251 83 552 41 * * Vesaliusweg 2-4 | e-Mail: filler-at-uni-muenster.de * * D-48149 Muenster Germany | filler-at-medsnt01.uni-muenster.de * ****** http://medweb.uni-muenster.de/institute/anat **********
Hello out there all you overworked & underpaid microscopy people. Hey, if you think you have it bad what about us poor students? Rice and lentils aren't exactly a good diet you know! Anyway - can anyone tell me if it is possible to carry out a decent X-ray microanalysis of an SEM sample of tissue? I am trying to see if 'calcified' tissue really does contain calcium, and can't find out whether the electron penetration into the tissue will be deep enough to let me know if there is any calcium in there or not. I mean, will I simply get info regarding the surface cells and not the actual interstitium (which is where the good stuff is hiding). I can't strip the cells off either as this is valuable PM material and I have to do routine SEM on it as well.... Thanks for the help in advance!
Alex ---------- Alex Black Department of Anatomy National University of Ireland, Galway Ireland
I have a Mitsubishi P78U, large format black & white video printer connected to my Kevex 8005. It works well and has been trouble free. The catch is, when
most video printers are working well, the quality (resolution/tonal range) is sub-standard for demanding SEM imaging. If you are capturing NTSC video, the
vertical resolution will never be greater than about 500 lines for the entire image. Most digital systems capture 1024 lines (pixels) or more. My polaroids are typically exposed at 2000 lines. Horizontal video resolution is related to system bandwidth, but is seldom greater than the vertical res.
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I need information about video printers for printing SEM images and need to contact sales representatives from various companies. Please contact me at 217-782-0898. Thanks. Donna Wagahoff SIU School of Medicine PO Box 19230 Springfield, Il 62794-1220
by generic.axs2000.net (8.9.1/8.8.8) with SMTP id IAA15279 for {Microscopy-at-MSA.Microscopy.com} ; Fri, 24 Jul 1998 08:56:38 -0400 Message-Id: {199807241256.IAA15279-at-generic.axs2000.net} To: MICROSCOPY BB {Microscopy-at-Sparc5.Microscopy.Com}
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Maoxu Qian wrote: =================================================== I'm looking for carbon EM calibration grid from SPI model 411CG-AB but couldn't find any information about SPI.Does anyone knows they phone # or www address? ==================================================== I think I can reply with some element of authority to this question!
The above mentioned part number was for our now discontinued product, the carbon composite grid. Now I am sure I will be corrected quickly should I be wrong, but the one person in the world who could make this product stopped making it some several years ago. So these grids are just not available any more. Note: If anyone has any that are not going to be used, we would love to have them back since we do still get requests for them. PS : It was never intended to have any applications as a "calibration grid".
We did acquire a pretty good idea of why one would want to use this grid and have offered on our website a list of substitute grid products. So far as we know, one or more of the substitute products seems to have been quite acceptable (or better or cheaper) in just about any application to our knowledge that has been presented.
You can find us from the information given below. Hope this helps!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
While some folk out there are talking knifemakers it might be useful for the LKB 7800 owners to have a look at our 1992 publication in J. Microscopy. In this short technical note we describe a series of simple modifications which transformed the performance of our two resident LKB's.
To quote from the conclusion;
'Analysis of the performance of an LKB 7800 series KnifeMaker revealed that its random failure was largely due to an accumulation of manufacturing tolerances, and of wear in the mechanism. After the playy was eliminated it was still necessary to adjust the cutter wheel positio accurately in order to obtain acceptable knives. With the modifications and adjustments outlined above, the number of acceptable knives that could be obtained from one strip of glass was improved byy at leasdt 100%.'
Don't throw it away - fix it !
Ref; Journal of Microscopy, Vol.168, Pt 1, October 1992, pp 111-114.
We had a, later, short paper on the advantages of applying tungsten coating to glass knives but I am sure that this is well documented.
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Private Bag X01, Scottsville 3209, KwaZulu-Natal, South Africa. Tel +27 (0)331 260 5155, Home +27 (0)331 962676 Fax +27 (0)331 260 5776 email: bruton-at-emu.unp.ac.za
This bouncing mail is originating at the University of Wisconsin, Madison, they were also the culprits the last time a loop got started about 2 years ago.
I am attempting to contact the individual, however, looking through the mail it appears that some of these messages are at least a year old and are being resent from slimbach-at- facstaff.wisc.edu for unknown reasons. It is NOT something that I have direct control over as it originates the University of Wisconsin. Neither of the two addresses I have identified as the source are current subscribers to the Listserver.
In any event please DONOT send any mail or reply to to the addresses below as it may be reflected to the server
My grad. school diet was PBR and pizza... all of the basic food groups...:-)
-- Begin original message --
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Hello out there all you overworked & underpaid microscopy people. Hey, if you think you have it bad what about us poor students? Rice and lentils aren't exactly a good diet you know! Anyway - can anyone tell me if it is possible to carry out a decent X-ray microanalysis of an SEM sample of tissue? I am trying to see if 'calcified' tissue really does contain calcium, and can't find out whether the electron penetration into the tissue will be deep enough to let me know if there is any calcium in there or not. I mean, will I simply get info regarding the surface cells and not the actual interstitium (which is where the good stuff is hiding). I can't strip the cells off either as this is valuable PM material and I have to do routine SEM on it as well.... Thanks for the help in advance!
Alex ---------- Alex Black Department of Anatomy National University of Ireland, Galway Ireland
-- End original message -- regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
A Philips EM 301 is available as a gift to anyone who is willing to take responsibility for removing it (including packing and shipping costs). The instrument has been in use until recently and is basically in good shape. It does have a couple of minor problems which would require a routine service visit. I do not have the exact age of the instrument but Philips stopped making this model about twenty years ago.
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 7 - October 15, 1998
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2050 (Includes room and board)
Application Deadline: August 4, 1998
Admission application and information: Carol Harnel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratio-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual research problems with the faculty.
Software for Logon and Booking of Electron Microscopes.
Lehigh University Microscopy Center wishes to replace the software and hardware it currently uses for logging on and off the microscopes, and for booking sessions, keeping records and so on.
Please let me know if you are aware of any good system for this purpose. Windows preferred but a Mac system would be quite acceptable.
(People with a long memory may recall that I made a similar posting a few years ago while still in my previous job at Illinois. The result of that posting was a lot of interesting information but we did not locate a system which really seemed to do everything one would wish. We hope there has been a lot of progress in the last few years.)
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
At 12:35 PM 7/24/98 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alex,
Rice and lentils will keep you alive forever, especially if washed down with good Irish stout.
I assume you are doing your x-ray analysis in an SEM? If so, you will get information both from the surface and from some distance down into the tissue, depending on the accelerating voltage (kV) you use. The higher the voltage, the deeper you penetrate the tissue.
Unless you are using a spot analysis mode, your x-ray spectrum will give you all the elements in the visual field of view. (This is apparently what you plan, since if you can't see the precise areas you want to measure, you can't put a beam precisely on them.) The quantity of calcium in the tissue will be a major factor in whether or not you can see it. If it is there in tiny amounts (say less than 1%), coupled with the fact that it may be under an unknown thickness of non-calcium bearing cells, you may not be able to see it at all.
Definitely don't coat your tissue with Au or Au/Pd. If coating is necessary, use carbon. Or, if you have variable pressure capability, try using the lowest pressure (1-5 Pa, for example) that allows you to stop the charging effects. You lose capability for high-resolution imaging, but for low mag work this may not matter.
Finally, if your system can do element mapping, it might be fun to run a map for calcium. It just might see it and give you a visual reference to its location. Can't hurt to try.
Good luck.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Corneliu Sarbu National Institute for Materials Physics POBox MG-7 R-76900 Bucharest-Magurele Romania
are being returned as undeliverable. Does anyone have another email address for him? Or, if Dr. Sarbu is reading this, can you confirm your email address please: csarbu-at-alpha1.infim.ro ?
Thanks, Phil
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net or poshel-at-hotmail.com
Thanks for the generous offers of information, not only by e-mail but by fax and telephone. I have passed your replies on to the prospective user of the instrument (Mark Schmitt).
Leonard R. Corwin Fort Dodge Animal Health Cyanamid Agricultural Research Center PO Box 400 Princeton, NJ 08543-0400 609-716-2278; fax 609-275-5239 corwinl-at-pt.cyanamid.com
I have a student in the lab who needs to double label some TEM sections. I've never done this before so I'm turning to y'all for expert advice. He needs to do two different kinds of double labelling. 1) Label with two antibodies both raised in mice. 2) Label with one mouse & one rabbit anitibody.
We're not clear as to how to proceed so if anyone out there has a protocol that they could let him use or at least tell us what to do, that would be great! At least we have the different size gold secondaries.
Seeing double in Berkeley,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207
As I told you in a message to this group a few weeks ago, we are looking = at new SEMs and would like to look at fixed but wet tissue i.e. Fallopian = tubes to see if their microvilli and cilia are in good enough condition to = continue with more preparation or in the case of hair cells in the rat = cochlea, to see if they are sufficiently exposed or if part of the lateral = wall still needs to be dissected away. Surely some people out there use variable pressure or environmental = microscopes to look at wet, fixed tissues without introducing artefacts = from drying or freezing. I have been in contact with several microscope = representatives and am getting conflicting answers about what will and = will not work and how long a wet sample can be looked at in the scope = without drying out. I need to hear from people who have these microscopes = and work with wet tissues and cold stages. This is a big event for us. Our current SEM scope is 23 years old, so we = don't get the chance to make a purchase like this often. We want to be as = well informed as possible and at this point, I am very confused. Please = help me answer these questions: How long can the wet tissue be looked at in either type scope? Is a cold stage necessary and how long can cold, wet samples be viewed? What adverse affects on the tissue result from the freezing temperature of = the cold stage? Can tissue be looked at in either of these scopes and then be processed = for high vacuum SEM and observed at high vacuum and high resolution? Thank you very much for any advice you can give.
Donna Wagahoff SIU School of Medicine PO Box 19230 Springfield, Il. 62794-1220 217-782-0898 fax 217-524-3227
Would the person who posted the notice about spare Philips 300 parts please resend it with a valid email address. I'm interested, but replys get returned undeliverable.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Alex Black wrote: ================================================== ........ can anyone tell me if it is possible to carry out a decent X-ray microanalysis of an SEM sample of tissue? I am trying to see if 'calcified' tissue really does contain calcium, and can't find out whether the electron penetration into the tissue will be deep enough to let me know if there is any calcium in there or not. I mean, will I simply get info regarding the surface cells and not the actual interstitium (which is where the good stuff is hiding). I can't strip the cells off either as this is valuable PM material and I have to do routine SEM on it as well.... ==================================================== Assuming you have a large enough sample that you are not going to get good data by SEM/EDS from the interior of the sample, there is a technique that can be used that involves embedding, sectioning and then plasma etching the resin away in a plasma etcher. The analysis is done by analytical TEM.
You can see the end result of that kind of a sample prep on our website URL http://www.2spi.com/catalog/instruments/etchers4.html
The technique is fairly straight-forward: You will have to embed your sample, thin section it and pick it up on a silicon dioxide filmed grid. You then expose the (unstained) section to an oxygen plasma in a small low power (not more than 100 watts) plasma etcher, to remove all organics, leaving the non-organics scattered around, showing the structure almost as a "ghost" image. However, the beauty of the technique is that you have removed the carbon that would contribute to high Bremmstrahlung background radiation, increasing greatly then the sensitivity for what it is that you do want to detect.
But this will enable you to get closer to if not actually on the target of your objectives.
After removal of the organics, and this is all supported on a SiO2 filmed grid, the inorganics essentially outline a "ghost" structure of the original cell structure. So to whatever degree you need maximum sensitivity, your sample is now in that form where you could have the best shot at detracting what you want to detect.
There have been publications using the technique, I just don't have any of them at my fingertips at the moment.
Disclaimer: SPI Supplies manufactures the plasma etchers and produces the silicon dioxide filmed grids on a custom basis all of which can be seen on our website below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We are in the process of developing just such a chart in conjuction with Kodak. Prototypes have already been constructed (including a gray scale, which should satisfy your black and white requirements).
I will forward your request; perhaps we can arrange for you to test one.
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I suggest using some tool software to do such thing. Assuming that you have proper hardware, you can:
1. Dupe the original disk by Disk-duping software(such as HD-Copy) as a backup; 2. Use a disk diagnostic software(such as Norton's Norton Disk Doctor) to repair the original disk; 3. Then try to dupe the repaired original disk again to get a usable one.
But from my point of view, it would be no problem if you have "copies of copies" that can work well. However, disk-duping software is recommended because some commercial disk may be encrypted between disk tracks, which can not be replicated by COPY command.
Regards.
Dr Yanping Zhou(Celia)
Electronic Microscopy Group Shanghai Institute of Ceramics Chinese Academy of Sciences 1295 Dingxi Road, Shanghai 200050 P. R. China } } } We have a NORAN (Tracor Northern) 5400 Series II EDS system (Model } Number TN-5402/BBAA) acquired in 1987. It does not have a hard drive and } operates off of two floppy drives. It requires 5.25" double sided, high } density floppy disks, the originals of which are no longer readable. They } have not been in use other than to be the source of copies made } periodically but apparently they have simply deteriorated with time. } We have copies of copies that are working pretty well but I'm becoming } concerned that they may crash/die and we'll have no good source from } which to make new copies. NORAN does not have replacement masters of this } vintage so I'm hoping to find someone who has original master disks in } good working order who would be willing to loan them to be copied. } } We need two disks of the set from the November, 1987 Release 1C. What will } work with our system is very specific and the original label on the master } disks reads: } } SERIES II SOFTWARE RELEASE 1C } NOV., 1987, TRACOR NORTHERN, INC. } } Of the disks in this release, we need: } } SQ/SSQ/PRZ/ZAF Master } NOT BOOTABLE } } and } } MSCAN2 Master } } } Thanks in advance for any help or suggestions you may have. } } ___________________________________________________________________________ } Barbara Reine, Botany Dept. Box 351330 } Univ. of Washington, Seattle, WA 98195-1330 } e-mail: reine-at-u.washington.edu; ph: (206) 543-1955 } ____________________________________________________________________________ } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu } }
While investigating a problem last week with the aluminum foil backing on our manufacturing plant's ceiling tiles, I thought it would be expedient to put a relevant question to an "Ask the Experts" group I found through SPI's website. To the metallurgists of that group I asked why fine abrasive action on most if not all metals creates a black substance. I asked if that would be the reduced form of the metal. The problem our plant is having is that in handling these tiles, fingers become black and then smudge the white surface. I received a short reply from the webmaster that my question was outside the scope of their interest and would normally be deleted without comment.
So, microscopists, does anyone have an idea about the cause of the blackening? Reflected light at 100x or so isn't very illuminating.
While investigating a problem last week with the aluminum foil backing on our manufacturing plant's ceiling tiles, I thought it would be expedient to put a relevant question to an "Ask the Experts" group I found through SPI's website. To the metallurgists of that group I asked why fine abrasive action on most if not all metals creates a black substance. I asked if that would be the reduced form of the metal. The problem our plant is having is that in handling these tiles, fingers become black and then smudge the white surface. I received a short reply from the webmaster that my question was outside the scope of their interest and would normally be deleted without comment.
So, microscopists, does anyone have an idea about the cause of the blackening? Reflected light at 100x or so isn't very illuminating.
Dear Dave, } } To the metallurgists of that group I asked why fine abrasive action on most } if not all metals creates a black substance. I asked if that would be the } reduced form of the metal...
No, it is not reduced metal--at least in the case of aluminum, which oxidises readily on exposure to air or water. } } So, microscopists, does anyone have an idea about the cause of the blackening? My guess is that the size and shape of the particles does not re- flect light. This guess could be tested by microscopic examination to characterize the particle size and shape. This problem may have already been solved by the particle experts (among whom McCrone Associates are frequent contributers to the list).
} Reflected light at 100x or so isn't very illuminating. } Due to the low albido. :-) Yours, Bill Tivol
On Thu, 23 Jul 1998 steve-at-facstaff.wisc.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Good morning Reynolds users } } In nearly 30 years of preparing lead citrate according to } Reynolds I have experienced no problems in using a stock } sodium hydroxide solution (1M) which was made up from pellets. } However, this solution must be freshly made up. Is it because I } always use Reynolds in its concentrated form that I do not } experience any problems? } } Rob } } } Robin H Cross } Director : EM Unit, Rhodes University, Grahamstown, South Africa } eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377 } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu } } There are probably a number of things you are doing RIGHT. However, if the NaOH solution is not of the correct normality (1.0), you will have a shift in staining. If it is far off to the alkaline side, you will get essentially no staining. Too "acid" and you get at first intense stain and then dumping. I have 4 years worth of data on this. If you get the correct normality with pellets, fine. Just remember the fact (listed in the original Reynold's paper that variations in pH cause variations in stain intensity.) All you need to know is this concept. How you get there is really immaterial. Bye, Hildy
P.S. I personally take the liberty of "diddling" with the pH of the Reynolds lead citrate. I know exactly how "acid" I can go to increase my intensity, if I want. I do this easily with a commercial source of NaOH which is titrated to the exact known normality.
On Thu, 23 Jul 1998 steve-at-facstaff.wisc.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Would freshly distilled water be equivalent to degassing via sonication ? } } Leo } } On Wed, 16 Apr 1997, Ian Montgomery wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } -----------------------------------------------------------------------. } } } } } } Good morning Reynolds users } } } } } } In nearly 30 years of preparing lead citrate according to } } } Reynolds I have experienced no problems in using a stock } } } sodium hydroxide solution (1M) which was made up from pellets. } } } However, this solution must be freshly made up. Is it because I } } } always use Reynolds in its concentrated form that I do not } } } experience any problems? } } } } } } Rob } } } } } Like Rob I've been making Reynold's lead citrate since I was a boy. } } I use NaOH pellets but degas the distilled water by sonicating it for a few } } minutes before making the stain. } } } } Ian. } } } } } } } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu } } Hi,
Guess what folks? I have been making Reynolds Pb citrate for 7 years without boiling, degassing water, etc. If you take care to chealate the Pb and citrate correctly, carbon dioxide just does not turn up as a problem. Now, end of Pb discussion! So long, Hildy Crowley
P.S. I never let students make Pb which I plan to use. I always make my own, because too many "inventions" without having studied the Reynold's paper lead to morasses.
I would need a 2.5 x microscope objective to fit a Diavert (a discontinued Leitz model; mine is about 12 years old). The tube length is 170 mm, with a DIN (45 mm) distance between slide and turret. If any of you on the list can supply this accessory, I would be interested in purchasing.
When a metal is finely divided the particles form a black powder. This is solely an optical problem, and not a materials problem. You might want to coat the aluminum with a thin coat of polymer to eliminate the abrasion.
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} While investigating a problem last week with the aluminum foil backing on our } manufacturing plant's ceiling tiles, I thought it would be expedient to put a } relevant question to an "Ask the Experts" group I found through SPI's website. } To the metallurgists of that group I asked why fine abrasive action on most if } not all metals creates a black substance. I asked if that would be the reduced } form of the metal. The problem our plant is having is that in handling these } tiles, fingers become black and then smudge the white surface. I received a } short reply from the webmaster that my question was outside the scope of their } interest and would normally be deleted without comment.
My understanding of this phenomenon (whereby any bright, shiny metal- including platinum-when ground finely enough gives a black rather than shiny powder) is not due so much by oxidation but to the fact that the metals have been ground in the micrometer range. Such particles no longer reflect the light in an ordered manner (like a mirror) back to the eye but in fact bend it in so many directions that the light is reflected mostly away from the eyes and gives the impression of absorbing rather than reflecting light. A good example of this is the photographic negative. The negatives have metallic (reduced) silver but it is of such a fine order (micrometers) that the negatives look very dark rather than shiny. When one rubs their hands over soft metals (like aluminum) you abrade off very fine flecks of the metal (micrometer range) that give your hands the blakened appearance. Harder metals (stainless steel for example) would do the same except that they do not abrade as easily.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Steve, Use either the Mg or Ca (*2-3-mM) in the fix and buffers (for maintaining membrane tonicity and keeping those myelin figures to a minimum). Also try some reduced osmium, it may make the membranes stand out, as will a good Ua/Pb (preferable citrate and nitrate) section stain.
I apparently accidently forwarded some junk email to the microscopy server. If you received any junk email from this address, MY SINCEREST APOLOGIES.
I realized I had accidentally hit the "forward" button on my email program, and hit the reset button ASAP. I guess I didn't catch it in time. I am not affiliated with this group that originally sent the mail, and I hate spam as much as the next guy. SORRY.
} I suggest that it may be necessary for } Nestor to configure the software that is used for this list to prevent } posting from non list members. } } What do other people think. Ian:
Well put, Ian. I agree. The sex post was the pits.
Blystone in Texas
Robert V. Blystone, Ph.D. {RBLYSTON-at-Trinity.edu} Professor of Biology Trinity University San Antonio, Texas 78212 210.736-7243 210.736-7229 FAX
I am looking for a person with extensive FIB and TEM sample prep experience
to fill a position at the IBM Almaden Research Center. The work would involve extensive collaboration with the groups at Almaden and in the IBM Storage Systems Division that are developing next-generation heads and disks, photoresists, low dielectric constant materials, and MRAM.
Candidates should have experience in as many of the following areas as possible: FIB, TEM sample prep - especially tripod polishing and micromanipulation, TEM, and SEM.
Please contact me directly. Submit your resume to: Robby Beyers K19/D1 IBM Almaden Research Center 650 Harry Road San Jose, CA 95120-6099
I am finishing my master (a pre-embedding study on serotonin) and before=20 i am trying to localise serotonin in a cnidarian tissu by post-embedding.= =20 My tissues are embedded in lowycril and are well preserved. I have no=20 problem in cutting them and I recolt them on nickel grids with formvar.=20 THE PROBLEM is the immuno in itself where I have way to much back ground=20 noise everywhere even on the formvar. My antibody is diluted 1 in 500 or=20 in 1000 or in 2000 for 4 hours and the secondary 1 on 75 for 1 hour,=20 centrifuged before use.=20 Those solution are rinsed and diluted in PBS .1M, BSA .5%, and .05%tween=20 20 (I have tried 1% cold water gelatin), and for the pre-incubation I=20 add10% normal goat serum. I simply transfer my grids from one droplet to=20 the other. As well, before preincubating, I rinse my tissues in water and= =20 sodium metaperiodate saturated for 30 min.
However whatever I try I don't get rid of this background noise. =20 HELP!!!!!!!!!
THANKS
NATACHA BENRIMOH UNIVERSIT=C9 DE MONTREAL DEPARTEMENT DE BIOLOGIE (514) 343-6111 #1052 benrimon-at-ere.Umontreal.ca
Dave, the black material is aluminum oxide that forms on all untreated aluminum, which is being wiped off by handling and transferred to the cosmetic part of the tile. Possible solutions would be another handling method or overcoating the aluminum somehow. We use disposable cotton glove liners to keep our hands clean(er), but that probably won't keep the tiles clean. Best of luck,
Vern
Vern Rieck Quality Engineer Aavid Thermal Products, Inc. rieck-at-aavid.com
-----Original Message----- From: David_R_Stadden-at-Armstrong.com [SMTP:David_R_Stadden-at-Armstrong.com] Sent: Monday, July 27, 1998 9:05 AM To: Microscopy-at-sparc5.microscopy.com Subject: Black metal particles
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While investigating a problem last week with the aluminum foil backing on our manufacturing plant's ceiling tiles, I thought it would be expedient to put a relevant question to an "Ask the Experts" group I found through SPI's website. To the metallurgists of that group I asked why fine abrasive action on most if not all metals creates a black substance. I asked if that would be the reduced form of the metal. The problem our plant is having is that in handling these tiles, fingers become black and then smudge the white surface. I received a short reply from the webmaster that my question was outside the scope of their interest and would normally be deleted without comment.
So, microscopists, does anyone have an idea about the cause of the blackening? Reflected light at 100x or so isn't very illuminating.
steve-at-facstaff.wisc.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear all } } I may be dragging the thread away from contamination, but I thought that } most electron microscope users avoided silicon based oils in diff pumps etc } } because they are extremely difficult to remove from the interior of an } electron microscope and any contamination would normally have electrical } insulating properties (catastrophic in an e.m.). Perhaps I am wrong but I } would welcome any comments. After all this is one of the reasons why } Santovac oils and their relatives became so popular (despite their costs). } } If I am labouring under a mis-apprehension then I apologise. } } Malcolm Haswell } University of Sunderland } UK } } All of the silicon based DP fluids I am aware of (which may not be all that } are on the market) will break down under an electron beam and deposit a } layer similar to glass on the nearest cool surface in the column. I don't } know of any EM manufacturers that would recommend their use because they } are almost impossible to remove if they do get in the column. We don't } even use silicon based fluids in our vacuum evaporator. } } K. A. Brackett, Ph.D. } TN & Assoc./USEPA } } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu
Malcom, I've been watching this thread with considerable interest, also. ETEC sold almost every one of its microscopes with Transene Vacoil (later Vacoil-S) which I believe is redistilled Dow-Corning 705. I have never had any problems with it in 21 years of servicing these instruments. Even a burped DP (and I've dealt with my share) has never been a problem beyond the fact that you have to clean everything. Many ETECs were sent out with the water-cooled baffle plumbed in just before the DP rather than before the power supplies. Those instruments will condense MP oil on their EDS detectors until they get replumbed. but silicone doesn't seem to present any problems.
Ken Converse owner Quality Images third party SEM service Delta, PA
steve-at-facstaff.wisc.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All, } I find oil on the EDX detectors in both my SEM's, but if you think of the } conditions inside the SEM, it is almost inevitable. The majority of the } molecules remaining in the vacuum of an average SEM at 10 -5 torr consists } of diffusion pump oil. This oil will gradually condense on all surfaces in } the SEM, but on the coolest surface first. In most SEM's this is the EDX } snout. One EDX manufacturer has solved the problem by warming up the snout } with a small heater. The oil just goes somewhere else, but if it forms a } thin film over all surfaces it is not so visible or worrying. } The cleaning method recommended by the manufacturer of my EDX's is to gently } run clean-grade iso-propanol over the snout, then allow to air-dry. They } used to recommend Freon, but that is no longer permitted or available. } Hope this helps, } Mary } } Mary Mager } Electron Microscopist } Metals and Materials Eng., UBC } 6350 Stores Rd. } Vancouver, B.C. V6T 1Z4 } CANADA } tel:604-822-5648, fax:604-822-3619 } e-mail: mager-at-unixg.ubc.ca } } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu
Mary, If you system is properly trapped, you shouldn't get any condensation, or at least very little. My experience has been that the oil is often mechanical pump oil, not DP oil, and may arise from valving over at too low a pressure, allowing considerable backstreaming of MP oil.
Ken Converse owner Quality Images third party SEM service Delta, PA
Our Hitachi 2460N variable pressure SEM has always been condensing oil on our detector snout. Typically, we have a large drop form every month. It has bothered me (a lot of things do these days) but everyone says that it is to be expected. I don't think so.
} If you system is properly trapped, you shouldn't get any condensation, } or at least very little. My experience has been that the oil is often } mechanical pump oil, not DP oil, and may arise from valving over at too } low a pressure, allowing considerable backstreaming of MP oil.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I suspect that much of your problem is due to the Formvar. Try doing the immunolabeling on naked gold grids. You can get more support by using a fairly high hexagonal mesh (300 mesh). Formvar is *very* sticky for antibodies and tracers!
If background is still high, try using a hypertonic saline rinse, about 5X normal (750 mM NaCl) every time you need to rinse. Just be sure to use a normal saline rinse or two before you continue with the labeling protocol.
Good luck, let us know how things work out!
Best regards,
Bob ***************************************** Robert (Bob) Chiovetti rchiovetti-at-aol.com E. Licht Company / 1-800-865-4248 Colorado/Utah/Wyoming/Arizona/ New Mexico/West Texas U.S.A. Representing Leica Since 1967 *****************************************
In my service days pretty well all the data that we gained by carrying ou= t an analysis on the detector oil droplet was that it is rotary pump fluid.= I should say even now I hardly see a microscope without this droplet on the=
EDX snout.
It was no good just changing to another fluid as the complete vacuum syst= em would be "contaminated" by the fluid. To start again required every part=
of the system to be cleaned, even rotary pump lines.
A system that has not been used could be cleaned up from day one using activated charcoal in a fore line trap. Try to help a used system withou= t a 100% clean up and you gain nothing.
Steve Chapman Senior Consultant =
Protrain for electron microscopy training and consultancy world wide see http://ourworld.compuserve.com/homepages/protrain
A student in my lab will be travelling to the UK from Australia to attend a conference next year in march. He would like to visit other labs who are working on citrus. He is making a study of oleocellosis a disorder related to damage to oil glands in the rind of the fruit during postharvest handling. He is currently undertaking a developmental study of oil gland development. Anyone able to help with interpretation of sectioned material would be ideal. He is preparing material for LM and TEM, hopes to use confocal for 3D reconstructions too.
replies to tknight-at-waite.adelaide.edu.au (or to me)
thanks
Meredith Wallwork
Dept Horticulture Viticulture and Oenology Waite Campus University of Adelaide South Australia AUSTRALIA
22 years ago I arrived at a new lab that had just bought a new Siemens Elmiskop102 TEM (their last model produced) and an ETEC Autoscan. Siemens, to the end recommended Silicon fluid for the pump system. By then it was "common" knowledge that silicone fluids caused column contamination problems. I changed both instruments fairly soon to Santavac 5. It is news to me that the Etec also contained silicone, it was just simpler to use one fluid in both instruments.
Point is some manufacturers are slow changing to new realities, and Old EMists spent horrendous time just keeping instruments in reasonable conditions. A lot more time was spent cleaning apertures and columns and adjusting the scope to be astigmatic twenty and more years ago. Instrument maintenance now is (comparatively) a snap. The ETEC is now quite geriatric, but if you are using a silicone based fluid it's time to change and lower maintenance time and increase average performance. It's easy to test if your vacuum fluid contains silicone; just put a drop onto a SEM stub, centre the stage and give it a few seconds analysis under an EDS. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************* www.proscitech.com.au *****
At 06:31 PM 7/27/98 -0600, John J. Bozzola wrote: } Our Hitachi 2460N variable pressure SEM has always been condensing oil on } our detector snout. Typically, we have a large drop form every month. It } has bothered me (a lot of things do these days) but everyone says that it } is to be expected. I don't think so. } } } If you system is properly trapped, you shouldn't get any condensation, } } or at least very little. My experience has been that the oil is often } } mechanical pump oil, not DP oil, and may arise from valving over at too } } low a pressure, allowing considerable backstreaming of MP oil.
We have a 2460N in our lab. Our standard procedure is to leave the system sucking air at 40 Pa during its idle times. This serves to maintain viscous flow conditions (proper term?) under which the oil is swept from the system. We have not yet had to clean our EDS detector during the several years we have been running it, and we still get good low energy peaks.
Meanwhile, we have a JEOL 840A with light element EDS at the other end of the hall and we see droplets form on it quite quickly. We lose more than half of our O intensity over a 6 month period and need to reclean.
BTW, I understand the viscous flow principle is the idea behind Ronald Vane's system for keeping columns clean. We haven't bought his system yet, but would definitely recomend considering it if the cleaning gets to be too bothersome.
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
A few days ago I posted a metallurgical question to the group in which I referred to an "Ask the Experts" group I'd found "through SPI's website". My message indicated a lack of success with that forum, but I want to make it clear that this was simply a link that I'd arrived at through SPI's pages and was not the SPI folks themselves. I've always been impressed with SPI (and certainly their web presence) and in no way wanted to reflect adversely on them.
Thanks to all who responded to my question; your feedback was a big help.
Dave Stadden Armstrong World Industries, Inc. Testing & Analysis Lab
Since the subject of Reynold's stain has come up recently I have a question. My stain performs well (I think) but with time more and more crystals precipitate out of solution onto the botton of the flask. Is this bad? Am I doing something wrong? The stain is made in the traditional way and is stored in a volumetric flask sealed with Parafilm in the 'fridge.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Has anyone tried using foreline traps to reduce oil buildup on EDS detectors? I have been thinking about trying one of the units made by Taylor Engineering that use copper maze absorbent. Stanley L. Flegler Center for Electron Optics Michigan State University
It seems that there is some confusion over my previous post. Let me try to clarify here.
At 12:59 PM 7/28/98 -0500, someone wrote: } I have a question in respect of your comment: } } Meanwhile, we have a JEOL 840A with light element EDS at the other end of } } the hall and we see droplets form on it quite quickly. We lose more than } } half of our O intensity over a 6 month period and need to reclean. } } Do you use the 40Pa flow of "air" (not N2?) on the 840A as well? If not, is } there any particular reason not to do it? If yes, why do you think it is } worse than your 2460N?
Our Hitachi 2460N is a "leaky" or environmental SEM (hence the N designation), while our JEOL 840A is conventional "high vacuum-only" SEM. Both are working just as designed in their day. So while we have a choice of pressure and gas for the sample chamber of the Hitachi, we have no such choice of vacuum level for the 840A. I rather wish we did, but at this time that would require a third party solution, hence my prior reference to XEI.
As far as why air and not N2, we normally run our 2460N chamber with helium for the reduced scattering that it affords at a given vacuum level. We simply pull the He hose from the SEM inlet filter barb when we are ready to leave the scope for the night. I suppose were we purists or wealthier we might just leave the He hooked up all the time.
I hope this helps clarify. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
After you make up your Reynolds, aliquot it into 1.5 ml eppy/microfuge = tubes and store in the frig. When you need it centrifuge a tube at = medium to high speed for 5 or so minutes and use. The solution when = stored in microfuge tubes will last several months. The lead citrate = solution from what I understand reacts with the glass somehow and ppt = outs.=20
Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
} Has anyone tried using foreline traps to reduce oil buildup on EDS } detectors? I have been thinking about trying one of the units made by } Taylor Engineering that use copper maze absorbent. } Stanley L. Flegler } Center for Electron Optics } Michigan State University
FWIW, we have a trap on the foreline of the rotary pump on our JEOL 840A for a couple of years. It may have helped some, but it has in no way stopped the oil condensation on our detector.
Is it possible to set up darkfield illumination without the expense of a whole different condenser? I have tried knife edge illumination (lying a sharp-edged on the (in my case) light source condenser on the base of the microscope, so that the knife (or card stock?) edge runs exactly through the axis of the light path). I thought of painting a black disk on the center of a transparent filter.
I'd be grateful for any useful suggestions. (I am using a Zeiss standard 16 with an na 1.35 condenser).
Alan -- Alan E. Davis Marianas High School (Science Department) AAA196, Box 10001 adavis-at-netpci.com http://www.saipan.netpci.com/~adavis Saipan, MP 96950 15.16oN 145.7oE GMT+10 Northern Mariana Islands
Do folk have strong opinions about the desirability of venting SEM chambers with either air (passed through a filter and desiccant column) or bottled nitrogen?
In the case of LEO SEMs with LaB6 filaments it strikes me as being an unnecessary expense to vent with bottled nitrogen as the column region is seldom vented. Also venting with bottled nitrogen adds complexity as precautions must be taken not to over pressurize the chamber and damage the EDS window.
Dear Sirs: I would like to apply for a post-doctor position in your laboratory. Now I have graduated as a Ph.D in Institute of Physics, Chinese Academy of Sciences. My study has focused on:
(1) The High Resolution Electron Microscopy investigation on the structure of NiFe/Mo and Fe/Mo multilayers (relating to GMR effect).
(2) The microstructure inverstigation on C ions-implanted silicon by HREM.
I would appreciate it very much if you would supply such a position for me in your laboratory.
} Since the subject of Reynold's stain has come up recently I have a } question. My stain performs well (I think) but with time more and more } crystals precipitate out of solution onto the botton of the flask. Is } this bad? Am I doing something wrong? The stain is made in the } traditional way and is stored in a volumetric flask sealed with Parafilm } in the 'fridge. } } Geoff McAuliffe, Ph.D.
---------------------------------------------------------------------------- -------- The problem is with atmospheric carbon dioxide. Every time you open the flask to remove some stain you introduce CO2. This reacts with the Lead Citrate in solution to give Lead Carbonate which is insoluble.
When you make up Lead Citrate it is important to use CO2 free water, either freshly produced from a high quality deioniser or boil distilled/deionised water and allow it to cool. Sonication will not remove dissolved CO2 as the gas has formed a low concentration of Carbonic acid in the water :- CO2 + H2O = H2CO3 ( H+ & HCO3 - )
To keep Lead Citrate, aliquot it into smaller containers like small vials or tubes and store in the fridge. Then use a new one each time you stain.
We have found that boil off from liquid nitrogen is by far the best venting material. It is dry, it is clean, it is cheal.We hane the whole lab' plumbed with a line going back to a 100litre Dewer which if I remember lasts for 3-4 weeks. You obviosly have to be careful with pressure. There is nothing like getting a 100lb SEM stage whislting out of the microscope and landing on your lap. Really brtings tears to your eyes.
Patrick Echlin Multi-Imaging Centre University of Cambridge.
On Wed, 29 Jul 1998, Trevor Sewell wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } } }
Either proposition will work. Both means require a bit of maintenance. I would be inclined to use nitrogen; it's drier and cleaner. The cost is mostly in the rental of the cylinder and not the gas. I used a full N2 cylinder to run some pneumatic EM valves and instrument backfilling. When the high pressure cylinder ran low, it was switched to serve for gas agitation during film development. The low pressure cylinder was always available to exchange for a full cylinder when required without interrupting EM operations. Over-pressurising is avoided by not relying on a timer to backfill the column but the use of a demand valve (from a diver shop) this type of valve delivers gas only while the diver, or the microscope "sucks". A "T" junction with a needle outlet near the cylinder could also be useful to dust-off particles from microscope parts etc. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
Do folk have strong opinions about the desirability of venting SEM chambers with either air (passed through a filter and desiccant column) or bottled nitrogen?
In the case of LEO SEMs with LaB6 filaments it strikes me as being an unnecessary expense to vent with bottled nitrogen as the column region is seldom vented. Also venting with bottled nitrogen adds complexity as precautions must be taken not to over pressurize the chamber and damage the EDS window.
Hi Trevor, This is a great topic for discussion as there are quite a few factors = that come into play. Basically the main criteria for any vacuum system is to keep the = atmosphere in it constant and clean. By venting and opening the door of = the chamber in a SEM to change samples obviously throws that criteria = out the window.=20 As is always the case with microscopy we have to make compromises. The = best solution would be to fit an airlock to the chamber. In this way you = would maintain a better vacuum, however then you are limited to sample = size and it's expensive to buy an airlock. The second option is to back fill with dry nitrogen gas. Nitrogen being = the gas that is most prevalent in the air that we breath and that vacuum = manufactures have therefore designed their vacuum pumps and gauges = around.=20 By back filling with dry nitrogen you are maintaining a fairly constant = atmosphere to pump after venting and changing the sample. Maintaining = the positive pressure in the chamber, as you open the door, keeps the = surrounding atmosphere out. Yes, the negative side to this is the danger = of putting to much pressure on the window of the ED detector and = therefore blowing it. This can be prevented by simply removing the latch = that hold the door closed.=20 The surface area of the door is much larger than the area of the window, = so the door will be forced open far before the pressure on the window of = the ED detector suffers. In fact on normal W filament systems where you = vent the entire column, I have seen the gun popping up and down due to = dry nitrogen backfilling, when the door latch is still on. No damage to = the ED detector. As mentioned the vacuum components are also suited to Nitrogen. Pirrani = and penning gauges are calibrated on Nitrogen pressures, so by using = nitrogen you have a better chance of attaining the same readings each = time you pump again. The Turbo molecular pump, in particular, prefers to = pump nitrogen and is thus more efficient. Water vapour is particularly = difficult for a turbo pump to pump. So if your EM is situated near the = coast or you have a lot of heavy breathers using the EM, the humidity in = the lab will change on an hourly, daily and seasonal basis. On a bad = day, with high humidity, the pumping speed of the chamber to reach VAC = OK status, could decrease in terms of 5 to 10 minutes from a good dry = day.=20
Thus in conclusion ( yes I eventually got there), to backfill with dry = nitrogen improves pump down speed, cleanliness of the system, stability = and repeatability of the required vacuum and prolonged life of the = vacuum pumps. This will be very helpful if you require high resolution = work or very accurate ED analysis from your system on a regular basis. = There is also no need to purchase an air lock for your system. On the down side, as there always is in microscopy, there is the cost of = the nitrogen, ensuring the door latch is not left on to prevent ED = window damage ( however I am not aware of anyone where this has = happened. ), changing samples in a hurry so as not to waste too much = Nitrogen and the hassle of making sure you have a supply of Nitrogen = handy all the time.=20 In my opinion, a SEM that is totally vented each time you need to change = samples, i.e. vent the column , gun and chamber, and is situated near = the coast or any other high humidity area, should definitely use dry = nitrogen backfilling. If you are simply venting the chamber but wish to do high resolution = work or stable ED analysis, I would still look at dry nitrogen = backfilling. If you are analysing or looking at really foul samples at low mag any = way, I would not bother.
That's it from me. I hope it helps and good luck.
Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
-----Original Message-----
Do folk have strong opinions about the desirability of venting SEM chambers with either air (passed through a filter and desiccant column) or bottled nitrogen?
In the case of LEO SEMs with LaB6 filaments it strikes me as being an unnecessary expense to vent with bottled nitrogen as the column region is seldom vented. Also venting with bottled nitrogen adds complexity as precautions must be taken not to over pressurize the chamber and damage the EDS window.
I've had some problems sending the microscope stage to positions already visited and stored using the ISIS "autorun" program. Indeed, I almost observe a "shift" of about 20 microns which means that the stage never comes back to its original position.
Suppose the stage is located by X1 Y1 and I then move to another location X2 Y2. If I then come back to X1 Y1, the image has shifted by about 20 microns to X3Y3. However if I change location and want to come back to X1Y1, the stage accurately comes back to X3Y3 each time...
Though I have never quantified it, using dry nitrogen seems to speed pumpdown somewhat, likely by minimizing water entry into the vacuum system. It is of no help, of course, when I get the ocassional specimen which outgasses significantly.
I have bottled nitrogen in the lab, but instead use the gas take-off provided on the LN2 cryo which is also in the lab.
Over pressure is not a problem so long as I don't hold the chamber door shut when venting....
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Do folk have strong opinions about the desirability of venting SEM chambers with either air (passed through a filter and desiccant column) or bottled nitrogen?
In the case of LEO SEMs with LaB6 filaments it strikes me as being an unnecessary expense to vent with bottled nitrogen as the column region is seldom vented. Also venting with bottled nitrogen adds complexity as precautions must be taken not to over pressurize the chamber and damage the EDS window.
(SMTP Gateway for microscopy-at-sparc5.microscopy.com); Wed, 29 Jul 1998 09:03:29 -0400 Message-Id: {199807291303.AA11170-at-gateway.ppg.com} Received: by gateway.ppg.com (Protected-side Proxy Mail Agent-1); Wed, 29 Jul 1998 09:03:29 -0400
----------
Position Open -- Immediately
Microscopy and Microanalysis Research Assistant
An immediate position is open for an SEM and TEM microscopist at the North Carolina State University Analytical Instrumentation Facility (AIF) as a retirement replacement.
Duties and responsibilities include: Teaching of laboratories; user training and assistance; and instrumentation development, modification and maintenance; and analysis of a wide variety of samples. An MS or equivalent experience in a materials related discipline along with three
or more years hands on experience with SEM and/or TEM is required. Required skills include: extensive hands on experience with SEM, TEM and
related techniques and accessories (e.g. specimen preparation and associated tools such as evaporators, etc.); teaching/user training; and
familiarity with modern electronics, computer systems and experience with vacuum systems.
Please send resume and three letters of reference to: Phil Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 118A EGRC; 1020 Main Campus Drive; Raleigh, NC 27695-7531 or email PRUSSELL-at-NCSU.EDU.
North Carolina State University is an Equal Opportunity, Affirmative Action Educator and Employer. Minority and Female Applicants are especially encouraged.
Inrormation about our lab can be found at http://spm.aif.ncsu.edu/aif/
-- Phillip E. Russell Professor, Materials Science and Engineering Director, Analytical Instrumentation Facility Box 7531, Room 318 EGRC 1010 Main Campus Drive North Carolina State University Raleigh, NC 27695-7531
Hi Alan, I have made perfectly acceptable darkfield-like micrographs by simply introducing a central stop in the light pathway at the condenser level exactly as you suggest. One time for a course I was taking, I used a dime to block the central part of the collumated light beam. This was a long time back so I don't recall exactly where I placed it but what-the-heck. Experiment a little. John
________________________ C. John Runions, Ph.D. Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
I agree that boil off from LN2 is the best backfill. I used to use it extensively when I was working with UHV systems. There is absolutely no H2O in it. It is the driest nitrogen that you can get and there is also very little O2. LN2 is usually very available in an electron microscopy lab that has EDS detectors on the instruments and it is cheap. However, you don't have to have the boil off from a large tank and the plumbing that goes with it or the overpressure danger. An open container of LN2 will suffice. It is at atmospheric pressure and therefore will not overpressure your vacuum system. Make sure that the tube that you stick in is long enough so that the liquid that is drawn out is converted to gas before entering the vacuum system. Also, make sure that the tube doesn't go to the bottom of your dewar, otherwise it will suck up debris.
Incidentally, for safety reasons, you should not use a heater in the bottom of a dewar that can be pressurized. You should also not pressurize a LN2 dewar with air. Liquid oxygen can form at the bottom of the dewar over time. But following this thread, avoid overpressuring the vacuum system. The open LN2 approach does that.
Nitrogen is pumped fairly well by all vacuum pumps, especially ion pumps. The rated pump speeds are all referenced to N2. There are other gases in filtered air that become significant partial pressures in the ultimate vacuum because of their lower pump speeds.
Use LN2, your vacuum system will love you for it.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
We have found that boil off from liquid nitrogen is by far the best venting material. It is dry, it is clean, it is cheal.We hane the whole lab' plumbed with a line going back to a 100litre Dewer which if I remember lasts for 3-4 weeks. You obviosly have to be careful with pressure. There is nothing like getting a 100lb SEM stage whislting out of the microscope and landing on your lap. Really brtings tears to your eyes.
Patrick Echlin Multi-Imaging Centre University of Cambridge.
On Wed, 29 Jul 1998, Trevor Sewell wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } } }
I've always recommended backfilling with N2 at about 1-2 lbs. over atmospheric pressure. I'm from the better-safe-than-sorry school and prefer to keep my chamber as clean as possible. As far as overpressurization, it hasn't been a problem with our Amray scopes because the stage/door is not locked to the chamber. At atmospheric pressure, there is sufficient leakage past the O-ring seal to prevent damage to our thin window EDS. I can only recall one incident in the past 11 years (more than 100 microscope-years) when N2 broke a window. That SEM was used by many people, was not well maintained, had undiagnosed problems with the N2 purge (more flow! decrease cycle time!) and a door that latched to the chamber.
My 2 cents
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
How often are you venting the chamber ? We have found on our LEO 440 that venting with nitrogen takes a long time (up to 30 minutes !!) (Maybe we're doing something wrong, Luc?). Although the pump down time decreases, you're probably going to lose more time venting than you'll gain with the decreased pump down time if you're changing samples often. Venting with nitrogen will keep the SEM cleaner, though, if time is not a problem.
Charles Bushell Mintek
} -----Original Message----- } From: Luc Harmsen [SMTP:anaspec-at-icon.co.za] } Sent: 29 July 1998 02:35 } To: 'Trevor Sewell'; 'MSA listserver'; 'MSSA listserver South Africa' } Subject: RE: Venting SEM chambers with bottled nitrogen or air } } Hi Trevor, } This is a great topic for discussion as there are quite a few factors that } come into play. } Basically the main criteria for any vacuum system is to keep the } atmosphere in it constant and clean. By venting and opening the door of } the chamber in a SEM to change samples obviously throws that criteria out } the window. } As is always the case with microscopy we have to make compromises. The } best solution would be to fit an airlock to the chamber. In this way you } would maintain a better vacuum, however then you are limited to sample } size and it's expensive to buy an airlock. } The second option is to back fill with dry nitrogen gas. Nitrogen being } the gas that is most prevalent in the air that we breath and that vacuum } manufactures have therefore designed their vacuum pumps and gauges around. } } By back filling with dry nitrogen you are maintaining a fairly constant } atmosphere to pump after venting and changing the sample. Maintaining the } positive pressure in the chamber, as you open the door, keeps the } surrounding atmosphere out. Yes, the negative side to this is the danger } of putting to much pressure on the window of the ED detector and therefore } blowing it. This can be prevented by simply removing the latch that hold } the door closed. } The surface area of the door is much larger than the area of the window, } so the door will be forced open far before the pressure on the window of } the ED detector suffers. In fact on normal W filament systems where you } vent the entire column, I have seen the gun popping up and down due to dry } nitrogen backfilling, when the door latch is still on. No damage to the ED } detector. } As mentioned the vacuum components are also suited to Nitrogen. Pirrani } and penning gauges are calibrated on Nitrogen pressures, so by using } nitrogen you have a better chance of attaining the same readings each time } you pump again. The Turbo molecular pump, in particular, prefers to pump } nitrogen and is thus more efficient. Water vapour is particularly } difficult for a turbo pump to pump. So if your EM is situated near the } coast or you have a lot of heavy breathers using the EM, the humidity in } the lab will change on an hourly, daily and seasonal basis. On a bad day, } with high humidity, the pumping speed of the chamber to reach VAC OK } status, could decrease in terms of 5 to 10 minutes from a good dry day. } } Thus in conclusion ( yes I eventually got there), to backfill with dry } nitrogen improves pump down speed, cleanliness of the system, stability } and repeatability of the required vacuum and prolonged life of the vacuum } pumps. This will be very helpful if you require high resolution work or } very accurate ED analysis from your system on a regular basis. There is } also no need to purchase an air lock for your system. } On the down side, as there always is in microscopy, there is the cost of } the nitrogen, ensuring the door latch is not left on to prevent ED window } damage ( however I am not aware of anyone where this has happened. ), } changing samples in a hurry so as not to waste too much Nitrogen and the } hassle of making sure you have a supply of Nitrogen handy all the time. } In my opinion, a SEM that is totally vented each time you need to change } samples, i.e. vent the column , gun and chamber, and is situated near the } coast or any other high humidity area, should definitely use dry nitrogen } backfilling. } If you are simply venting the chamber but wish to do high resolution work } or stable ED analysis, I would still look at dry nitrogen backfilling. } If you are analysing or looking at really foul samples at low mag any way, } I would not bother. } } That's it from me. I hope it helps and good luck. } } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za } } } -----Original Message----- } From: Trevor Sewell [SMTP:sewell-at-uctvms.uct.ac.za] } Sent: 29 July 1998 09:18 } To: Microscopy-at-sparc5.microscopy.com } Subject: Venting SEM chambers with bottled nitrogen or air } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } }
Another risk to the EDS thin window from venting which I haven't seen mentioned here is ballistic damage -- particulates getting blasted off the sample and punching a pinhole in the window. We see this problem probably more often than implosion failures. Be aware of the geometry of the venting flow pattern relative to the detector if you're considering a customized venting system and analyze powders or other particulates.
The whole concept behind Darkfield is that you eliminate the background light by putting some sort of device in the front focal plane of the condenser (where the iris is located) so that the light approaching the objective comes in at such a high angle, it misses the objective entirely. You can certainly make your own darfield patch stop ... and your message indicates that your thinking is headed in the right direction. Here are the instructions:
1. Set the microscope up for regular brightfield, Koehler illumination
2. Remove the eyepiece and look down into the "back focal plane" of the objective (all the way down the tube). You should be able to see the image of the condenser iris. (You can test by watching as you adjust the iris setting)
3. Open the condenser iris until it just disappears from the objective back focal plane.
4. Very carefully (so as not to disturb the iris setting), remove the condenser from its mount. Measure the diameter of the opening. That is the diameter of the circle you will need to pain on the center of your transparent "filter"
Note that the condenser iris setting is very dependent on the numerical aperture of the objective. This home-made approach will work best with low mag objectives (4x, 10x, 16x).
The alternative to going to all this trouble is to buy a simple "patch stop" from Zeiss. (Other suppliers, like Edmund Scientific at www.edsci.com, or Carolina Biological may also carry them). These patchstops are made from shim metal and work well just by opening the condenser iris fully then carefully inserting the stop just shy of the iris. They may need a bit of tape (smooth black electrical tape works well) to hold them in place but be careful not to get the adhesive on the blades of the iris.
Another variation on this theme is Rheinberg illumination. Instead of blocking the center with black (and getting darkfield), you can block it with a colored filter. The center of the filter will color the background and the ring around the edge will color the features in your specimen. EX: blue center stop with yellow ring produces yellow objects against a blue background. This technique works beautifully 10x objectives and with moderately scattering objects like filamentous mold and fairly flat diatoms. We offer sets of Rheinberg filters (I think there are over a dozen rings per sheet)for modest price. See our web site: { {http://www.MME-Microscopy.com/education} . While you are there, check out my book, "Optimizing Light Microscopy for Biological and Clinical Laboratories". It has lots of little experiments which might be of value to both you and your students.
adavis-at-netpci.com wrote: } } Is it possible to set up darkfield illumination without the expense of a whole different condenser? I have tried knife edge illumination (lying a sharp-edged on the (in my case) light source condenser on the base of the microscope, so that the knife (or card stock?) edge runs exactly through the axis of the light path). I thought of painting a black disk on the center of a transparent filter. } } I'd be grateful for any useful suggestions. (I am using a Zeiss standard 16 with an na 1.35 condenser). }
Alan:
I have used a cork-borer to cut dark field stops of various sizes out of heavy black paper. A machinist could punch out metal stops of various sizes from a sheet of aluminum.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
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One possibility is the use of a few drops of mineral oil (like PCR oil or similar) on the top of each aliquot - this protects the solution from carbon dioxide.
Michal
Michal Jarnik Lab of Structural Biology Research, NIAMS, National Institutes of Health, Bethesda, Maryland 20892. Ph: (301) 435-2587
Hi, I am trying to determine the theoretical minimum to the spot size of a laser (gaussian) beam at the focus of a 100X/0.85 (air) microscope objective for uniform illumination of the objective. Does the diffraction limit criteria for incoherent illumination also apply in the case of coherent gaussian beams?
It has been asked if crystals at the bottome of Reynold's Pb citrate is undesirable.
Answer: Yes. Something is wrong. Probably the chealation is not complete, allowing the crystals to settle out. The shaking (by hand and not by stirrer, rotator or shaker) and inversion for 25 minutes is critical. It is boring, but necessary. Pb is best not kept in glass. Transfer it to a disposable plastic syringe. It is worth looking into, since there is a problem lurking somewhere which might result in sudden, unexpected loss of grids. Bye, Hildy Crowley {hcrowley-at-du.edu}
Trevor, We vent all columns with nitrogen that is syphoned off a liquid nitrogen storage tank. This nitrogen gas is also run through a tube with a drying agent to make sure it doesn't pick up moisture from the lines.
We have scuba valves on the lines that will close as soon as you reach atmospheric pressure. This eliminates the problem of accidentally pressurizing the microscope column.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Do folk have strong opinions about the desirability of venting SEM chambers with either air (passed through a filter and desiccant column) or bottled nitrogen?
In the case of LEO SEMs with LaB6 filaments it strikes me as being an unnecessary expense to vent with bottled nitrogen as the column region is seldom vented. Also venting with bottled nitrogen adds complexity as precautions must be taken not to over pressurize the chamber and damage the EDS window.
I would appreciate comments for my guidance.
Best regards,
Trevor Sewell
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for the following problem always indicate your position as to direction and when you record where you were and wish to return to there always return in the same direction. If you head to it in a differnt direction you will also add an amout of space that includes the backlash in the stage.... Does any of this make sense? Let me know if I need to phrase it in a differnt manner. Thanks Ed Sharpe, Archivist SMECC
P.S. It is an old trick I learned when I was learning about tool and die making as an expansion of my mind. Dear all,
I've had some problems sending the microscope stage to positions already visited and stored using the ISIS "autorun" program. Indeed, I almost observe a "shift" of about 20 microns which means that the stage never comes back to its original position.
Suppose the stage is located by X1 Y1 and I then move to another location X2 Y2. If I then come back to X1 Y1, the image has shifted by about 20 microns to X3Y3. However if I change location and want to come back to X1Y1, the stage accurately comes back to X3Y3 each time...
Something is wrong with your system. For some reason, your flow is down. It shouldn't take any longer to vent with N2 than with air. Check the you still have gas, that you don't have any kinks in your supply hose, and that your valve is open. As a check, take the hose off at the inlet to your vacuum system. Moisten your lips and have the gas flow over your lips. You should just feel a slight cooling sensation. That is a good flow rate for venting a system.
How often are you venting the chamber ? We have found on our LEO 440 that venting with nitrogen takes a long time (up to 30 minutes !!) (Maybe we're doing something wrong, Luc?). Although the pump down time decreases, you're probably going to lose more time venting than you'll gain with the decreased pump down time if you're changing samples often. Venting with nitrogen will keep the SEM cleaner, though, if time is not a problem.
Charles Bushell Mintek
} -----Original Message----- } From: Luc Harmsen [SMTP:anaspec-at-icon.co.za] } Sent: 29 July 1998 02:35 } To: 'Trevor Sewell'; 'MSA listserver'; 'MSSA listserver South Africa' } Subject: RE: Venting SEM chambers with bottled nitrogen or air } } Hi Trevor, } This is a great topic for discussion as there are quite a few factors that } come into play. } Basically the main criteria for any vacuum system is to keep the } atmosphere in it constant and clean. By venting and opening the door of } the chamber in a SEM to change samples obviously throws that criteria out } the window. } As is always the case with microscopy we have to make compromises. The } best solution would be to fit an airlock to the chamber. In this way you } would maintain a better vacuum, however then you are limited to sample } size and it's expensive to buy an airlock. } The second option is to back fill with dry nitrogen gas. Nitrogen being } the gas that is most prevalent in the air that we breath and that vacuum } manufactures have therefore designed their vacuum pumps and gauges around. } } By back filling with dry nitrogen you are maintaining a fairly constant } atmosphere to pump after venting and changing the sample. Maintaining the } positive pressure in the chamber, as you open the door, keeps the } surrounding atmosphere out. Yes, the negative side to this is the danger } of putting to much pressure on the window of the ED detector and therefore } blowing it. This can be prevented by simply removing the latch that hold } the door closed. } The surface area of the door is much larger than the area of the window, } so the door will be forced open far before the pressure on the window of } the ED detector suffers. In fact on normal W filament systems where you } vent the entire column, I have seen the gun popping up and down due to dry } nitrogen backfilling, when the door latch is still on. No damage to the ED } detector. } As mentioned the vacuum components are also suited to Nitrogen. Pirrani } and penning gauges are calibrated on Nitrogen pressures, so by using } nitrogen you have a better chance of attaining the same readings each time } you pump again. The Turbo molecular pump, in particular, prefers to pump } nitrogen and is thus more efficient. Water vapour is particularly } difficult for a turbo pump to pump. So if your EM is situated near the } coast or you have a lot of heavy breathers using the EM, the humidity in } the lab will change on an hourly, daily and seasonal basis. On a bad day, } with high humidity, the pumping speed of the chamber to reach VAC OK } status, could decrease in terms of 5 to 10 minutes from a good dry day. } } Thus in conclusion ( yes I eventually got there), to backfill with dry } nitrogen improves pump down speed, cleanliness of the system, stability } and repeatability of the required vacuum and prolonged life of the vacuum } pumps. This will be very helpful if you require high resolution work or } very accurate ED analysis from your system on a regular basis. There is } also no need to purchase an air lock for your system. } On the down side, as there always is in microscopy, there is the cost of } the nitrogen, ensuring the door latch is not left on to prevent ED window } damage ( however I am not aware of anyone where this has happened. ), } changing samples in a hurry so as not to waste too much Nitrogen and the } hassle of making sure you have a supply of Nitrogen handy all the time. } In my opinion, a SEM that is totally vented each time you need to change } samples, i.e. vent the column , gun and chamber, and is situated near the } coast or any other high humidity area, should definitely use dry nitrogen } backfilling. } If you are simply venting the chamber but wish to do high resolution work } or stable ED analysis, I would still look at dry nitrogen backfilling. } If you are analysing or looking at really foul samples at low mag any way, } I would not bother. } } That's it from me. I hope it helps and good luck. } } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za } } } -----Original Message----- } From: Trevor Sewell [SMTP:sewell-at-uctvms.uct.ac.za] } Sent: 29 July 1998 09:18 } To: Microscopy-at-sparc5.microscopy.com } Subject: Venting SEM chambers with bottled nitrogen or air } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } }
I too have been planning to go to the ICEM in Can Cun, Mexico, and have been concerned about matters of safety there. However, I have talked with several people who have been there recently, and a couple who go there often on business, and have been informed that the area near the Conference Center, where the meetings will be held, is populated primarily by hotels and other commercial institutions, and is highly 'touristized', and so is probably no more 'dangerous' than any other major tourist center. However, I have received several warnings about being careful with luggage, wallets, briefcases, etc. in airports in Mexico, and on tours outside the Can Cum tourist area.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Barbra Foster's advice on dark field is exactly right. However, there is another way. Dark field is one of the few contrast methods in light mocroscopy that does not use Koehler illumination. You can place an opaque circular stop in the middle of the top lens of your condenser and adjust the height of the condenser while observing a focused specimen until a dark field image is formed. (Note, I do not mean on top of a flip-in lens if there is one but on top of the main condenser lens with any flip-in lens out.) Start with a stop about 4 to 5 mm in diameter. You may have to experiment with the size. This will work for 10 X and 20 X objectives. The thinner and more opaque the stop the better.
Bob
_____________________________________________________________ C. Robert Bagnell Jr., Ph.D., Res.Assoc.Prof. Microscopy Services Laboratory Department of Pathology & Laboratory Medicine CB #7525 UNC-CH, Chapel Hill, N.C. 27599 ph 919-966-2413 fx 919-966-6718
Dear Frank, } } I've had some problems sending the microscope stage to positions } already visited and stored using the ISIS "autorun" program. Indeed, I } almost observe a "shift" of about 20 microns which means that the } stage never comes back to its original position. } Our positioning program on the IVEM takes backlash into account and always drives to a set distance at a given orientation with respect to the end position. The last adjustment is then made slowly and always in the same direction. One chooses an initial position and goes away then returns several times until the stage returns reliably to the chosen position. Thereafter, moving to new positions and returning to old ones is automatic. I don't know who wrote our program (or yours), so I can't say how you might incorporate this procedure. Good luck. Yours, Bill Tivol
Regarding the discussion on darkfield imaging, I sometimes image specimens in pseudo darkfield using a 10X objective combined with the phase-3 (100x) phase ring of our condenser assembly. The darkfield image is by no means perfect but is often useful nonetheless. The ph3 condenser ring directs a cone of light through the specimen, which passes to the outside of the objective (darkfield). Put a small beaker of coffee on top of a slide to see this for yourself. Specimen visualization is dependent on light refracted into the objective by the specimen.
I can feel some of you nodding your heads...try it, you might be surprised!
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
Hi Jayesh, The eq for diffraction limited spot size is D(lim)=2.44 x Lambda x focal length/D(a) where D(a) is the diameter of the beam incident the lens. Remember this is the "theoretical diffraction limited spotsize" (assumes paraxial approximation) For input beams that are not collimated, but have a full divergence angle of A, the focal spot diameter will be approximately D(lim)=fA Most optical equations assume a paraxial approximation for the incident rays & a TEM 00 (Gaussian) E-field. There is also the issue of chromatic aberrations which would limit your (broad band) spot size. Diffraction limited spot size is wavelength dependent. Even if you were provided with a diffraction limited spot size or spot resolution by your objective manufacture, somewhere in fine print it is probably at a specified wavelength or 2. There is also an inverse dependence on the diameter of the beam incident on the lens. If the manufacture specified a Dlim.spot size , they are probably assuming the "clear aperture " of the objective is illuminated. The focal length is also wavelength dependent although a 100x objective hopefully has some chromatic correction. Your laser beam may not fill the clear aperture or it may, this brings in a another problem. Theoretically a Gaussian beam (E-field) extends forever. In practice to minimize Fresnel fringing (rings) your clear aperture should be 5x the beam radius at the lens. Fresnel fringes are an optical interference effect. Coherent light is required for optical interference to occur. Coherence is generally ignored in broad band applications. FYI, In reality a incandescent source has a coherence length of ~10um. Another point worth making. Unless you have a many 100K or few M$ lens, you theoretical diffraction limited spot size is no better than 1/2um. A microscope with a ~$100, 40x objective can resolve a 1um period. Using a lower mag. objective will give you a greater depth of field. With a 40X obj. depth of field is still only a few um. Where do you measure the beam waist? The focal plane is not the focal length from the end of the objective & it is not the working distance (although it will be close). It is the focal length measured from a plane known a A2H2 (a principle plane). Your objective supplier may be able to supply the approximate distance from some location on the objective. Since I have no idea what laser you are using, I'll point out that optical coatings can be damaged by lasers. If you have enough power, a focused beam will ionize the air (usually limited to pulsed laser applications). Laser beams can burn dust on the lens & in doing so damage the AR coatings. Also since you are interested in small spot sizes. UV(} 300nm) is heavily attenuated by most non reflective objectives.
hope this helps,
Bruce Brinson Rice U.
Jayesh C. Jasapara wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } I am trying to determine the theoretical minimum to the spot size of a } laser (gaussian) beam at the focus of a 100X/0.85 (air) microscope } objective for uniform illumination of the objective. Does the diffraction } limit criteria for incoherent illumination also apply in the case of } coherent gaussian beams? } } Thanks, } Jayesh
The big problem here is water vapor. Nitrogen backfill is primarily used to prevent the incursion of water vapor into the chamber since most vacuum systems have a hard time dealing with it.
The use of a desiccant column is well advised for any instrument. In the long run however, there are two problems. The first is the regeneration of the desiccant. You have to pay attention and bake the water out of it at regular intervals. The second is a long term accumulation of desiccant dust. Over time, the desiccants appear to produce a very fine dust that will eventually clog the chamber vent valve, resulting in a service call.
Nitrogen systems can produce their own problems. The need to have a constant source of nitrogen can be eased by using the vapor from a large storage dewar if liquid nitrogen is always available in the lab due to EDS or other cryogenic systems. Generally, the nitrogen is cheap but the demmurage charges for the dewar are not. If you use a little more nitrogen per month, the cost is minimal if you already have the need for the large dewar.
The savings is basically in time. The primary volume in an SEM vacuum system is in the sample chamber. It is not system cleanliness that is in question here, it is basically the time involved in pumping the chamber to required levels. If one can exclude water vapor from entering the sample chamber during sample changes, there will be an appreciable improvement in pump down times. A well airconditioned room is a start, desiccants next and a nitrogen backfill is best. Airconditioning is a necessary first step, since a warm and humid environment can also cause other problems such as the condensation of water on the exterior of water cooling lines used for diffusion pump and electronics cooling.
Most system contamination comes not from gases not pumped out, but from air leaks and pump oils. By the time a system reaches operational vacuum, the overwhelming majority of environmental gases, including water vapor, will be gone.
} Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Plenty. You should first determine if the mechanical assembly is operating properly. When dealing with the magnifications involved in microscopy, backlash is a primary concern. This refers to the difference in position caused by approaching the desired position from different directions. In many optical microscope stages, there are no anti-backlash measures taken to reduce the effect. The gear teeth, or leadscrew threads, are allowed to rest against the opposing, driving teeth or threads. If the teeth or threads are not tightly meshed, then the position attained can vary by the amount that the teeth or threads don't mesh.
In your case, you are probably coming from a low position to the first, X1 Y1, position. Then going upward to the X2 Y2 position. Don't be surprised when you move downward back to the X1 Y1 position to find the ultimate position offset by the amount of backlash. As a simple expedient, move back to X1 Y1 by first dropping well below that position, and moving back up to it. If you always come into a given position in the same direction, you should see a great improvement.
In the case of a simple stage movement, without anti-backlash provisions, the best you can do is clean the gears and leadscrews carefully, lubricate them properly and ensure that they are adjusted to provide a good tooth/thread mesh without binding.
Anti-backlash gears and leadscrew followers are essentially devices that bias the gear/leadscrew to always favor one side of the tooth/thread through spring action. Regardless of the direction of movement, they should always 'push' up against the same side of the tooth or thread. Basically they are comprised of two gears or leadscrew followers (nuts) that are spring loaded in opposing directions. One gear or follower is attached to the mechanism being moved, the other simply 'floats' to provide the bias.
This problem is referred to as 'repeatability' and can often vary a great deal from the accuracy spec that most people pay attention to. In the case of anti-backlash movements, proper cleaning and lubrication should be all that is needed to maintain a good repeatability. If not, then the separate parts should be inspected for wear.
} Dear all, } } } I've had some problems sending the microscope stage to } positions already visited and stored using the ISIS "autorun" } program. Indeed, I almost observe a "shift" of about 20 microns } which means that the stage never comes back to its original } position. } } Suppose the stage is located by X1 Y1 and I then move to } another location X2 Y2. If I then come back to X1 Y1, the image } has shifted by about 20 microns to X3Y3. However if I change } location and want to come back to X1Y1, the stage accurately } comes back to X3Y3 each time... } } } Any suggestions? } } F Allen R. SamŞŞŞŞŞŞŞŞpson Advced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
I have been responsible for a number of demonstration laboratories in the=
past and I have always insisted upon using a dry gas rather than dirty ol= d (English) wet air.
Why?
1. Pump down time is improved considerably to ~50% in some cases, but = it takes months for this to happen.
2. Column cleanlyness improves so if you work at low kV (and you all should try it) you will get a longer life from your column and its apertures between cleaning.
3. Contamination is the biggest killer of high resolution microscopy and=
this is also dramatically reduced.
Regards
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the studies of the three-dimensional structures in biology, medicine and material sciences. The three-dimensional analysis and representation are crucial steps to assess the information. These conferences are the most efficient meeting points between developers and users in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation and in particular to the role of the green fluorescent protein.
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
For about five years we have vented our Zeiss DSM 962 and TEM 900 using a nitrogen generator. It remains permanently switched on and takes nitrogen from the atmosphere; purifies it to remove moisture and other gases then transfers it to a tank to provide gas on tap. The main advantage of this system is that it is almost maintenance free and avoids the problem of changing cylinders. The venting time on the DSM is around 1minute 30 and the vacuum levels on both instuments are very good.
Frieda Christie, Electron Microscopist, Scientific & Technical Services, Royal Botanic Garden, 20A Inverleith Row, Edinburgh, EH3 5LR
I have been very surprised by the tone of several postings on the subject of safety in Cancun. Let us be quite clear about this. A visit to Cancun will be a lot safer than a visit to New York or any other major US city. If your bags are mishandled or stolen, it is much more likely to occur in Miami than in Cancun.
Cancun is a fine place. It is a great resort, suitable for an excellent vacation. The Congress promises to be most successful.
No one should be deterred from attending by these implied slights.
Alwyn Eades . Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I could not agree more! I have not visited Cancun, but I have been to Cozumel a couple of times and it is no different to any other Carribean vacation destination. In fact it is a custom built vacation city, built with the foreign visitor in mind. I am not going to the meeting for financial reasons, but no one should be deterred from visitng for safety reasons!!!
At 7:44 AM -0500 07/30/1998, Alwyn Eades wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
________________________________ Please note the new Area Code ________________________________
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
I forgot to add a neat part of the experiment. If you rotate the objectives out of position (you may have to remove one to get enough room) then place a piece of white paper (like a business card)on edge on the stage, over the light as it emerges from the condenser, you can see how you have changed it by any of the adjustments we discussed. Try it first with plain Koehler. Open and close the condenser iris and you can see how that affects the angle of light approaching the sample. Then close the condenser iris and open and close the field iris. You will see that it does, indeed, control the size of the illuminated field. Then try the experiment with (a) your darkfield patch stop and (b) Rheinberg filters. It's a neat demonstration.
If you have access to the High School shop and if they make screwdrivers as part of their curriculum, have them cut some of the handle stock (usually slighly turbid yellow plastic) into about 1" lengths. Oil one of these little "columns" to a microscope slide with a drop of immersion oil (Nujol or mineral oil from the pharmacy works well, too). Try the same experiments.
{bold} {italic} {bigger} {bigger} MME {/bigger} {/bigger} {/italic} {/bold} is America's first national consortium dedicated to
customized on-site training in all areas of
microscopy, sample preparation, and image analysis.
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At 10:14 AM 7/29/98 -0400, Barbara Foster wrote:
} } } }
{excerpt} ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Alan,
The whole concept behind Darkfield is that you eliminate the background light by putting some sort of device in the front focal plane of the condenser (where the iris is located) so that the light approaching the objective comes in at such a high angle, it misses the objective entirely. You can certainly make your own darfield patch stop ... and your message indicates that your thinking is headed in the right direction. Here are the instructions:
1. Set the microscope up for regular brightfield, Koehler illumination
2. Remove the eyepiece and look down into the "back focal plane" of the objective (all the way down the tube). You should be able to see the image of the condenser iris. (You can test by watching as you adjust the iris setting)
3. Open the condenser iris until it just disappears from the objective back focal plane.
4. Very carefully (so as not to disturb the iris setting), remove the condenser from its mount. Measure the diameter of the opening. That is the diameter of the circle you will need to pain on the center of your transparent "filter"
Note that the condenser iris setting is very dependent on the numerical aperture of the objective. This home-made approach will work best with low mag objectives (4x, 10x, 16x).
The alternative to going to all this trouble is to buy a simple "patch stop" from Zeiss. (Other suppliers, like Edmund Scientific at www.edsci.com, or Carolina Biological may also carry them). These patchstops are made from shim metal and work well just by opening the condenser iris fully then carefully inserting the stop just shy of the iris. They may need a bit of tape (smooth black electrical tape works well) to hold them in place but be careful not to get the adhesive on the blades of the iris.
Another variation on this theme is Rheinberg illumination. Instead of blocking the center with black (and getting darkfield), you can block it with a colored filter. The center of the filter will color the background and the ring around the edge will color the features in your specimen. EX: blue center stop with yellow ring produces yellow objects against a blue background. This technique works beautifully 10x objectives and with moderately scattering objects like filamentous mold and fairly flat diatoms. We offer sets of Rheinberg filters (I think there are over a dozen rings per sheet)for modest price. See our web site: { {http://www.MME-Microscopy.com/education} . While you are there, check out my book, "Optimizing Light Microscopy for Biological and Clinical Laboratories". It has lots of little experiments which might be of value to both you and your students.
Well, I hope you enjoy Cancun, but...... Several years (approx 5? ?) the American Chemical Society held a meetin= g in Mexico city. An ACS member was killed in his hotel room when he surpri= sed a thief when he returned unexpectedly to his room. I just read about a M= exican police officer who kidnapped three teenagers, took them to a police sta= ble, raped the two younger girls (while the old one locked herself in a room= to prevent the assault). The three escaped later. All the travel protect= ion web sights have lists an arm long discussing what not to do in Mexico (like= don't take any taxi that comes along). Let me see, I don't speak the language (my fault) I don't have family connections to their society I can't carry a effective means of self defense I don't understand the society while I am far from wealthy, to them I am extremely wealthy. Take some advice from my friend Mas Ayoob: "Don't go were you are not w= anted."
jae5-at-lehigh.edu on 07/30/98 08:59:43 AM To: microscopy-at-Sparc5.Microscopy.Com -at- INTERNET cc:
I have been very surprised by the tone of several postings on the subje= ct of safety in Cancun. Let us be quite clear about this. A visit to Ca= ncun will be a lot safer than a visit to New York or any other major US city= . If your bags are mishandled or stolen, it is much more likely to occur = in Miami than in Cancun.
Cancun is a fine place. It is a great resort, suitable for an excellen= t vacation. The Congress promises to be most successful.
No one should be deterred from attending by these implied slights.
Alwyn Eades . Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Dear all, =20 =20 Thank you for the many replies I have received regarding the backlash=20 problems I recently communicated to the list. From the replies I=20 received, it appears that some of you associated the problem with the=20 ISIS software mentioned in the E-mail and more particularly the=20 AUTORUN application. However, the above software has been performing=20 very well, i.e showing excellent repeatability (double clicking on=20 stored points does recall the correct coordinates). The problem as=20 some of you correctly pointed out, is associated with the lack of=20 backlash when carrying out stagee rotations. When using translations=20 only to go from 1 point to the other (and later come back to those=20 positions) everything is fine, but as soon as a rotation is introduced= =20 the repeatability is affected.Unfortunately there is nothing I can do=20 about it other than using translations to move from one point to the=20 other or always approaching a position from the same direction =20 F
I've found this works fine with a 10X objective and the 40X phase ring, or a 20X objective and the 100X phase ring. The image-especially the 10X-40X one was as good as a true dark-field image.
Phil
} } Regarding the discussion on darkfield imaging, I sometimes } image specimens in pseudo darkfield using a 10X objective } combined with the phase-3 (100x) phase ring of our } condenser assembly. The darkfield image is by no means } perfect but is often useful nonetheless. The ph3 condenser } ring directs a cone of light through the specimen, which } passes to the outside of the objective (darkfield). Put a } small beaker of coffee on top of a slide to see this for } yourself. Specimen visualization is dependent on light } refracted into the objective by the specimen. } } I can feel some of you nodding your heads...try it, you } might be surprised! } } Doug } ---------------------- } Douglas R. Keene } Associate Investigator } Shriners Hospital Microscopy Unit } 3101 S.W. Sam Jackson Park Road } Portland, Oregon 97201 } 503-221-3434 } DRK-at-shcc.org
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net or poshel-at-hotmail.com
We are talking Cancun here! It is a tourist destination, harming tourists is extremely bad for business. I cannot believe you are comparing Mexico City with Cancun. Mexico City is just like New York! Or come to think of it any other huge urban sprawl!
Please stop this rampant racism and just go and enjoy the conference!
At 9:43 AM -0400 7/30/98, FRANK KARL wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Note new Area Code (734)
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
I quite agree with Alwyn. I have been to Cancun and Cozumel numerous times and have never had any problems regarding safety or theft. I have walked the streets well into the night (something I would never do in most american cities) and not had any problem.The only inconvience I have ever incurred were people trying to sell me timeshares, even then, a couple of polite but firm "No Gracias" usually sends them on their way. While there I highly recommend visiting the Mayan ruins of Chichen Izta and Cubo. Take a bus tour though, driving can be dangerous as the roads are narrow and not in the best of shape. I once had a very large bull charge our rented jeep while doing 80 kph (the jeep not the bull). If you SCUBA dive, Cozumel has some of the best sites in the western hemisphere. On a side note I always drink bottled water while there, no matter where I am staying.
(sigh) It's a shame my company won't send me to meetings outside of the USA.
Chuck Mass Bio-Medical Imaging Wyeth-Ayerst Research 641 Ridge Road Chazy NY 12921 massc-at-war.wyeth.com
Dear Frank, } } [skip] When using translations } only to go from 1 point to the other (and later come back to those } positions) everything is fine, but as soon as a rotation is introduced } the repeatability is affected.Unfortunately there is nothing I can do } about it other than using translations to move from one point to the } other or always approaching a position from the same direction } The same technique which eliminates backlash from translations will also work for rotations (i.e., always approach the new orientation from the same direction), and the additional complications due to cross- talk between the translational and rotational backlashes can also be corrected by always doing one first, then doing the other, and approach- ing from the same directions. A possible protocol would be to rotate until one is a few degrees cw of the desired orientation, next translate to a point (delta-x, delta-y) from the desired position, then progress ccw to the desired orientation, and finally translate to the desired posi- tion. The important thing here is to choose the signs of delta-x and delta-y so that the movement in the (-delta-x, -delta-y) direction will not reintroduce rotational backlash. One may have to repeat the final adjustments in certain cases to be sure that the backlashes have, indeed, been properly taken up. Yours, Bill Tivol
We are looking to expand our capabilities here at The Jackson Laboratory so that we can image either entire mice or living, attached parts (legs, ears, etc.) One of our options may be MRI. So we're looking to find out who makes MRI machines, what they cost, if they do whole, small animals, what level of resolution can be achieved (i.e. enough to detect auto-immune induced degeneration), what qualifications are needed to operate this, etc. For people who may already be doing this, we'd be interested in your experience with these techniques.
We also think that ultrasound may be another way to do imaging. Or there may be methods out there that we don't know of yet. If anyone has any experience or ideas or knows of someone (investigator or manufacturer) who is doing this, I'd really like to hear from them.
Thank you!
Lesley Bechtold Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor ME 04605
{fixed} I see we have mixed feelings about safety in Can Cun. I have travelled to many places in Mexico, both on business and pleasure. In particular I have visited the hotel region of Can Cun where ICEM is being held and I would agree with Alwyn Eades and John Mansfield that this is is a no more dangerous place that any tourist area in the U.S. However, if you propose to visit some of the marvellous tourist attactions on the Yucatan peninsula such as Tolumn, Merida, Chichen Itza etc etc and you are not going on an organized trip {bold} {underline} beware {/underline} {/bold} , especially if you do not speak Spanish and are not in tune with the Mexican mentality. Police bribery for alleged traffic offences is not uncommon (if you have rented a car), petty overcharging for gas and other items is common and crime can be problem. My advice if you are not travelling in a large organized group is:- don't carry lots of cash, just a credit card or two and carry them in a safety holster or belt and brush up on your Spanish. Can Cun and its surroundings are magnificent (if a bit hot in summer). Take appropriate precautions and enjoy yourself.
Alan Fox
Director, Center for Materials Science and Engineering
} While there I highly recommend visiting the Mayan ruins of Chichen Izta } and Cubo. Take a bus tour though,
I have been to Chichen Itza and it was well worth the visit, we flew there from Cozumel. The tour of the ruins and the round trip airfare was about $130 each five years ago. Highly recommended. Clmb the pyramid if it is still open, they were going to close it was the tourists were wearing it away!
} On a side note I always drink bottled water while there, no matter where I am staying. Oh, yes this is a MUST! Even with the most careful precautions in Acapulco we all got Montezuma's Revenge from something we ate or drank. Things to note:
1. Ice, dont use it unless you are sure it was made with purified water. You can usually tell by the shape of the cubes, if it looks like the machine packed ice you get in the US then it is usually OK, but I would always ask to make sure. If the ice is home made it may have been made with tap water and then you will be in trouble, your system is not used to the flora and fauna of that part of the world!
2. Dont eat unpeeled fruit like apples, grapes, pears, etc. they may have been washed in tap water.
3. Dont eat salad vegetables (lettuce, cucumber, tomatoes, etc.) as they are likely to be washed in tap water too. This is difficult, as many foods come with a salad garnish and simply removing it is not always enough. We had shark quesadillas in Acapulco that I think were our downfall as they were accompanied by shredd lettuce as a garnish.
4. Brush your teeth with bottled water too, the little you may swallow while doing this may make a difference.
I know this all sounds like overkill, but it can save you many hours in the little room! Drink bere or pop it is usually cheap and plentiful in Mexico and the beer is good too!
John M.
Note new Area Code (734)
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Most of the people have been describing the boil off from large tanks outside (where the plumbing is required) or the large upright (5 feet high - 2.5 feet diameter). Both of these have valves to take off the vapor above the liquid. This pressure can be quite high and they also have pressure relief valves and safety blowout plugs.
You are correct about what I was saying. I used a small dewar that is used for absorption pumps which are basically styrofoam. You can use the styrofoam that is used to protect acid bottles when they are shipped or your dewar that you use to fill the EDS detectors. (If your are in a bind, put a tube into the EDS detector dewars.) If you have an open container, just use Tygon. If you have a narrow neck, use a copper or stainless steel tube to go into the tank so that it won't go hard if it is bent and attach the Tygon tube onto the warm end of the tube. Now you have to clear the line of the air in it just like any other line that you attach to a vacuum system. Just after you put the tube into the LN2, there will be blow out at the other end of the tube until the temperature equilibrates. Position that end over the inlet to the vacuum system to flush out the inlet also. While the gas is flowing, put the tube on the inlet and that will give you a fairly clean line.
It doesn't matter what diameter(s) that you use, just length. About 8-10 feet is good enough. If you use a stainless steel tube to insert in the LN2, it will blow gas a little longer than if you use copper because of its lower thermal conductivity.
I agree that boil off from LN2 is the best backfill. I used to use it extensively when I was working with UHV systems. There is absolutely no H2O in it. It is the driest nitrogen that you can get and there is also very little O2. LN2 is usually very available in an electron microscopy lab that has EDS detectors on the instruments and it is cheap. However, you don't have to have the boil off from a large tank and the plumbing that goes with it or the overpressure danger. An open container of LN2 will suffice. It is at atmospheric pressure and therefore will not overpressure your vacuum system. Make sure that the tube that you stick in is long enough so that the liquid that is drawn out is converted to gas before entering the vacuum system. Also, make sure that the tube doesn't go to the bottom of your dewar, otherwise it will suck up debris.
Incidentally, for safety reasons, you should not use a heater in the bottom of a dewar that can be pressurized. You should also not pressurize a LN2 dewar with air. Liquid oxygen can form at the bottom of the dewar over time. But following this thread, avoid overpressuring the vacuum system. The open LN2 approach does that.
Nitrogen is pumped fairly well by all vacuum pumps, especially ion pumps. The rated pump speeds are all referenced to N2. There are other gases in filtered air that become significant partial pressures in the ultimate vacuum because of their lower pump speeds.
Use LN2, your vacuum system will love you for it.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
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(412) 820-8651 (office) (412) 820-8161 (fax)
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We have found that boil off from liquid nitrogen is by far the best venting material. It is dry, it is clean, it is cheal.We hane the whole lab' plumbed with a line going back to a 100litre Dewer which if I remember lasts for 3-4 weeks. You obviosly have to be careful with pressure. There is nothing like getting a 100lb SEM stage whislting out of the microscope and landing on your lap. Really brtings tears to your eyes.
Patrick Echlin Multi-Imaging Centre University of Cambridge.
On Wed, 29 Jul 1998, Trevor Sewell wrote:
}
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-----------------------------------------------------------------------. } } Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } } }
At 10:20 AM 7/30/98 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hmmmm..... People are discussing safety at a destination they are unfamiliar with and are labelled racists.
Who is overreacting?
Just a thought.
Randy Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
I've followed with interest the debate on safety in Cancun. I've spent many years doing fieldwork in remote parts of Latin America and spent some time in such cities as Caracas, Montevideo, Buenos Aires, Santiago, and Lima. Without exception, I feel safer in those localities than most US cities. While in Peru this summer, I heard about:
A mass execution of two gang members and three innocent bystanders in Tacoma, WA, the nearest city to where I live.
A brutal beating on a rural highway in the small twon I live in.
Murder and mayhem in the US Capitol building.
During other trips to that region, I've read in their papers about the senseless murder of European tourists in Miami - on several occasions.
Most Latinos I've spoken with in South America express deep concern about visiting the gun-toting, violence prone society north of the Rio Grande.
I'd say that your chances of encountering crime and personal harm probably diminish when you leave US borders. So - go to Cancun and feel safer than at home!
A friend of mine needs to part with his FF instrument. Please direct your inquiries directly to Dr. Ip. (S. Samuelsson)
CRESSINGTON CFE-50 FREEZE ETCH UNIT This is an oil diffusion pumped unit with full freeze etch capability. It has a rotary stage. Deposition of Pt/C is carried out using EH5 electron beam guns and is controlled by an EB1202PC/C electron beam controller and an MTM22 quartz crystal monitor. This CFE-50 was installed in 1992 but has made less than 30 runs during its life because our laboratory changed research projects shortly after the unit was purchased. We will consider any reasonable offer.
Contact: Wallace Ip, Department of Cell Biology, University of Cincinnati College of Medicine, PO Box 670521, Cincinnati, OH 454267-0521. Tel. (513) 558-3614 Fax (513) 558-4454 Email: wallace.ip-at-uc.edu
} } I have been very surprised by the tone of several postings on the subject } of safety in Cancun. Let us be quite clear about this. A visit to Cancun } will be a lot safer than a visit to New York or any other major US city. } If your bags are mishandled or stolen, it is much more likely to occur in } Miami than in Cancun. } } Cancun is a fine place. It is a great resort, suitable for an excellent } vacation. The Congress promises to be most successful. } } No one should be deterred from attending by these implied slights. } } Alwyn Eades
I'd like to add that I've been all over the Yucatan (Chichen Itza, Uxmal, Merida, Coba, Tulum, etc, and have never encountered anything but exciting sights, fun, friendliness, good food, etc., etc. We WERE attacked by a swarm of beautiful butterflies on the road to Coba, tho...
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
We are using a Leica CPC unit for freezing bare grid technique. I was wondering if anyone else has one of these units yet and can give us some advice on it.
We have several problems:
The release mechanism for the plunger is threaded for what looks like a place to attach a photographic cable release. This would be great as otherwise one needs three hands to operate the unit and normally these type of units have a footswitch to control the drop of the plunger into cryogen. So, we purchased an air release (the kind with a hollow rubber ball at the end which you squeeze to release the shutter), and this turns out to be the wrong thread. I was wondering if Leica have made a part for this, and whether it is included in the price of the unit, but the local office in Canada did not know.
I think its a pretty crappy release mechanism for a 16,000 dollar unit! (it does not even include a binocular). All of the homemade ones I have seen (including the one my workshop made for me in Oxford) had an electronic release (they even made a 12 volt solenoid for me, so as to avoid sparks and ignition hazard when using ethane as a cryogen). You can buy an electronic footswitch in Tandy (UK) or Radio shack (N. America) for a few pennies.
By the way, the metal mirror attachment comes as an accessory which will cost you an extra 4000 dollars- it isnt included with the standard unit, so be warned! The brochure gives you the impression that this thing will do every type of freezing possible. Make sure they include the cost of the pump to keep the thing filled from a Dewar too, it won't work without this as its not intended to be able to fill it manually!
A big problem we have is ice contamination. The unit relies on a nitrogen "inert gas" atmosphere" from boil off. Problem with this is that you get "snow" falling into the unit from the turbulent air on top. You cant fill it with enough liquid to get your grids quickly under liquid where they are safe.
When using cryogens such as liquid ethane, the thing has to use a heater to keep the ethane from going solid. This is all very well, and it works, but it causes the ethane to evaporate very quickly, so instead of having to thaw the stuff with a metal rod, you continually have to top it up. Also you can't adjust the distance of the grid forceps from the surface of the ethane, the movement of the grid from start to stop is fixed, you cant change the length of movement or the distance it plunges, as it is the grid is dangerously close to the cold area and can pre-freeze if you are not careful. We will probably drill a few more holes so the thing can at least be mounted at different heights, but this wont give continuous adjustment. Also the forceps clamp does not hold very well and you are limited to forceps which have straight sides and are the same length as the paur supplied with the unit. A lot of the specimen holder appears to be mad out of heavy gauge hypodermic needles.!?!
So, as long as you don't need the automatic fill capability, you can make something yourself out of a styrofoam box and bottle cap and a plunger arm can be made in any workshop very cheaply- all it has to be is a cylindical metal rod running on roller bearings. A couple of clamps and stops control the height and distance of throw. Use a metal rod to thaw the cryogen. after freezing a specimen, top up so the ethane cup is under liquid nitrogen, and it will remain ice free for some minutes until you need to use it again, then just pour iff the excess nitrogen and warm the ethane with the metal rod (I mounted mine on a rubber cork, for insulation). You can use the same container for metal mirror, all you need is blocks of metal and if gravity is not fast enough for you, attach some heavy duty rubber bands.
Dr Timothy F. Booth Canadian Food Inspection Agency National Centre For Foreign Animal Disease Suite T2300 1015 Arlington St. Winnipeg Manitoba R3E 3M4 CANADA http://www.hc-sc.gc.ca/main/lcdc/web/bmb/fedlab_e.html#toc email tbooth-at-em.agr.ca Tel 204 789 2022 Fax 204 789 2038
There is nothing wrong with NYC either. It's a nice city; however, you just must be aware of where you are and remember not to act carelessly with yourself or your belongings. I reccommend being aware of your surroundings and of scammers that you often find in tourist traps and airports etc., but *please*, if you live your whole life in *fear* you'll have no fun at all.
Andrea
On Thu, 30 Jul 1998 10:20:30 -0400 jfmjfm-at-engin.umich.edu (John F. Mansfield) wrote:
} } } } I have been very surprised by the tone of several postings on the subject } } of safety in Cancun. Let us be quite clear about this. A visit to Cancun } } will be a lot safer than a visit to New York or any other major US city. } } If your bags are mishandled or stolen, it is much more likely to occur in } } Miami than in Cancun.
I'd like to add that I've been all over the Yucatan (Chichen Itza, Uxmal, Merida, Coba, Tulum, etc), and have never encountered anything but exciting sights, fun, friendliness, good food, etc., etc. We WERE attacked by a swarm of beautiful butterflies on the road to Coba, tho...
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Hi Bruce, Thanks for your help. I happen to be a novice in the field of microscopy - my specialization being femtosecond laser systems. I have a couple more questions for you. The only information I have on the objective is that it is a 100X and the N.A. is 0.85 . I am using this objective to focus down femtosecond pulses centered at 800nm wavelength and when I measure the spot size in the focal plane using the edge scan method, it measures to be about 350nm for full aperture illumination. Is this small a spot size possible for a wavelength of 800nm?
The location of the focal plane has been located accurately by looking for the steepest rise for the edge scan. The beam hitting the objective is not collimated by a telescope - it is a diverging beam.
The way I calculate the focal length of the objective from the given N.A. is by the following formula : f = D(a)/(2*tan(arcsin(N.A.))) = 1.55mm, where D(a) is the diameter of the incident beam on the lense. I am not sure about the validity/accuracy of this expression. Could you tell me how good or bad this expression works out to be?
If I use the formula assuming the paraxial case approx. holds, then the theoretical limit to the spot size is 500nm. But from the diverging beam formula Dlim = fA, I get a smaller theoretical spot size of about 395nm. This is closer to what I measure experimentally. The objective I use is of good quality (from Zeiss ) - so it should be corrected for chromatic aberration. Also the mean power being focussed is low - and so there is no danger of damaging coatings.
Thanks, Jayesh
} Hi Jayesh, } The eq for diffraction limited spot size is D(lim)=2.44 x Lambda x focal } length/D(a) } where D(a) is the diameter of the beam incident the lens. Remember this is } the "theoretical diffraction limited spotsize" (assumes paraxial } approximation) } For input beams that are not collimated, but have a full divergence angle } of A, the focal spot diameter will be approximately D(lim)=fA } Most optical equations assume a paraxial approximation for the incident } rays & a TEM 00 (Gaussian) E-field. There is also the issue of chromatic } aberrations which would limit your (broad band) spot size. Diffraction } limited spot size is wavelength dependent. Even if you were provided with a } diffraction limited spot size or spot resolution by your objective } manufacture, somewhere in fine print it is probably at a specified wavelength } or 2. There is also an inverse dependence on the diameter of the beam } incident on the lens. If the manufacture specified a Dlim.spot size , they } are probably assuming the "clear aperture " of the objective is illuminated. } The focal length is also wavelength dependent although a 100x objective } hopefully has some chromatic correction. } Your laser beam may not fill the clear aperture or it may, this brings in } a another problem. Theoretically a Gaussian beam (E-field) extends forever. } In practice to minimize Fresnel fringing (rings) your clear aperture should } be 5x the beam radius at the lens. Fresnel fringes are an optical } interference effect. Coherent light is required for optical interference to } occur. Coherence is generally ignored in broad band applications. FYI, In } reality a incandescent source has a coherence length of ~10um. } Another point worth making. Unless you have a many 100K or few M$ lens, } you theoretical diffraction limited spot size is no better than 1/2um. A } microscope with a ~$100, 40x objective can resolve a 1um period. Using a } lower mag. objective will give you a greater depth of field. With a 40X obj. } depth of field is still only a few um. } Where do you measure the beam waist? The focal plane is not the focal } length from the end of the objective & it is not the working distance } (although it will be close). It is the focal length measured from a plane } known a A2H2 (a principle plane). Your objective supplier may be able to } supply the approximate distance from some location on the objective. } Since I have no idea what laser you are using, I'll point out that } optical coatings can be damaged by lasers. If you have enough power, a } focused beam will ionize the air (usually limited to pulsed laser } applications). Laser beams can burn dust on the lens & in doing so damage the } AR coatings. Also since you are interested in small spot sizes. UV(} 300nm) } is heavily attenuated by most non reflective objectives. } } hope this helps, } } Bruce Brinson } Rice U. } } Jayesh C. Jasapara wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hi, } } I am trying to determine the theoretical minimum to the spot size of a } } laser (gaussian) beam at the focus of a 100X/0.85 (air) microscope } } objective for uniform illumination of the objective. Does the diffraction } } limit criteria for incoherent illumination also apply in the case of } } coherent gaussian beams? } } } } Thanks, } } Jayesh } } } }
Massimo, I totally agree! I was just about to voice the same opinions. I too am surprised (appalled!) to find such narrow-mindness and xenophobia among supposedly open-minded "scientists". These folks have been looking down their very narrow little tubes far too long and have severe micro-tunnel vision.
} Introduce oil? Don't we have enough trouble with section contamination? } If you chealate the two chemicals properly, you will not have to worry } about carbon dioxide. Keep your solution in a plastic syringe! Drive all } the air out of it. It will keep for years in the refrigerator. Put a } clean 0.22 micrometer filter on the syringe before use. Wet the filter } with the solution before forcing a drop out which is to be used for a } grid. } } Hildy
I switched from Reynold's recipe to Haniachi's version (Hanichi et al. 1986. A stable lead modification of Sato's method. J. Electron. Microsc. 35: 304-306) several years ago and have been quite pleased with the results. Although I could get Reynold's to work for me, I was never 100% successful and my student's grids were rarely precipitate free. Now, we use a single 30-60 sec stain with the calcined lead and don't even bother with a uranium secondary contrast (radioactive wastes have become a thorny issue here). I work mostly with Spurr's embedded plant material but we also do Epon embedded animal tissues for EM class; both have nice contrast.
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
I am not going to Cancun, but rather wish I was. However, I was just asked by a friend who has a son leaving for Mexico, what is needed by way of proof of identity for travel to Mexico? I know that neither passport or visa is necessary for travel to Canada. Is the same true of our southern neighbor?
At 02:52 PM 7/31/98 +1000, you wrote: } Dry oxygen maybe a lesser consideration, however, when the filament chamber } is to be vented, it's even more important to wait until the filament has } cooled. Exposure of a hot filament or LaB6 cathode to N2 is not a good idea, } exposure to O2 is worse. } Cheers } Jim Darley } ProSciTech Microscopy PLUS
Hmmm. We vent our Hitachi with W filament with He (because of less scattering effect compared to N2). Should we be concerned about any reaction between the hot filament and the He? I would presume none. How about thermal shock effects? Anybody care to comment?
No offense intended, but this discussion is getting way too anal. I'm not sure what 'scattering' effects you believe helium will improve. If you are referring to electron scattering during operation, helium is probably harder for normal vacuum systems to pump out then nitrogen, so the gain you are seeking more than likely isn't there. I know of no practical difference between venting with any dry gas.
Conductive heat dissipation is always much faster than convective. In the two or three seconds it takes you switch the accelerating voltage off and to hit the vent button, the majority of the heat built up in the small mass of a tungsten filament will probably be conducted off through the filament posts and electrical contacts in the gun. As the backfill gas works its way into the gun, the remainder of the heat will be removed rather slowly since the leak provided by a normal vent valve is rather small, taking 30 seconds or more to completely vent.
I have never seen or heard of anything other than very loose anecdotal evidence for increases in cathode life dependent on venting methods. At the most, I'll accept customers turning the filament drive down before turning off the accelerating voltage and venting the instrument. Anything more than that is simply unnecessary. It is far more important to be cognizant of small vacumm leaks, particularily in areas that could bleed into the gun area. Air leaks in the gun during operation are extremely damaging.
} At 02:52 PM 7/31/98 +1000, you wrote: } } Dry oxygen maybe a lesser consideration, however, when the filament } } chamber is to be vented, it's even more important to wait until the } } filament has cooled. Exposure of a hot filament or LaB6 cathode to } } N2 is not a good idea, exposure to O2 is worse. Cheers Jim Darley } } ProSciTech Microscopy PLUS } } Hmmm. We vent our Hitachi with W filament with He (because of less } scattering effect compared to N2). Should we be concerned about any } reaction between the hot filament and the He? I would presume none. } How about thermal shock effects? Anybody care to comment? Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Note: Entries for the 14th ICEM Photomicrograph Exhibit will be accepted through August 15th.
ICEM 14 - Cancun PHOTOMICROGRAPHS ON DISPLAY
There will be a Photomicrograph Exhibit held in conjunction with the 14th International Congress on Electron Microscopy in Cancun this August 31 - September 4. If you are attending the meeting, please consider bringing up to 3 micrographs for inclusion in a Photomicrograph Exhibit. From this exhibit, images will be selected for inclusion in a post conference publication. See details below and if you have questions contact either the meeting headquarters in Mexico City (info below) or myself.
Photomicrograph Exhibition
The 14th ICEM will include a micrograph exhibition. The organizers' intentions are to provide a showcase for outstanding micrographs displaying a combination of art and science.
The exhibit is open to all forms of microscopic imaging. A panel of judges will select entries based on artistic merit for inclusion in a post-congress publication of images. Submitted micrographs much be accompanied by a brief description, not only of their scientific content, but also of their technical aspects. If images have been digitally processed or altered, the digital processing should be described as well. All submitted micrographs will be displayed although entries not regarded by the judges as artistically significant will be excluded from the publication of images. Please read the following rules carefully:
Only registered attendees at the 14th ICEM are eligible to submit micrographs.
Any individual may submit up to three micrographs.
Entries must be 27.5 cm x 35.0 cm, may be mounted vertically or horizontally, and must be affixed to a stiff lightweight support, such as 10 mm foam board. Micrographs may either be flush mounted or have borders so long as the overall dimensions of the entry are 27.5 cm x 35.0 cm.
Entries must be brought to the meeting and mounted on the display boards by the entrant or his/her delegate. Velcro will be provided. Entries must be mounted between 12:00 and 17:00 on Monday, August 31st and those micrographs not selected for publication must be removed between 12:00 and 15:00 on Friday, September 4th. Micrographs remaining on the display boards after then will be discarded. Micrographs selected for publication will not be returned to the entrant and will become the property of the 14th ICEM.
Selected micrographs will be incorporated into a post-congress publication of images. Those selected for inclusion will be announced during the Thursday evening reception.
To enter: Send your name, address, telephone and fax numbers, and email address plus a description of 200 words or less for each entered micrograph (maximum of 3) to: Secretariat 14th International Congress on Electron Microscopy Amsterdam 46-202 Col. Hipodromo Condesa C.P. 06100, Mexico, D.F. MEXICO Phone: (525) 553-4507; Fax (525) 553-4500 email: icem-at-icem.inin.mx
Entry information must be received by August 15, 1998. Entries will be acknowledged promptly. Do not send micrographs, you must bring them or have them brought to the Meeting.
Meeting Office staff will print your description(s) in standard form and prepare them to be mounted along with your micrograph(s) at the Meeting.
___________________________________________________________________________ Barbara Reine, Botany Dept. Box 351330 Univ. of Washington, Seattle, WA 98195-1330 e-mail: reine-at-u.washington.edu; Phone: (206) 543-1955; Fax: (206) 543-3262 ____________________________________________________________________________
I should have mentioned that our Hitachi is a 2460N low vacuum SEM. During operation we He instead of air or N2 in order to reduce scattering effects at the 40 Pa of atmosphere that we maintain in order to cancel charging on our specimens. There was no intent of trying to reduce scatter by residual gas at 10-5 torr levels since He might have displaced heavier molecules.
I was intrigued that a side benefit of our He use might be increased filament life since the hot filament would not be exposed to N2 with its detrimental effects. A coworker likes to wait until the filament cools for a bit before venting the chamber (the gun chamber always gets vented with the specimen chamber). However, I had been supposing that thermal issues might be insignificant as you appear to corroborate. Since we just had a W filament go 441 hours even with sample changes about every 2 hours, I suppose that the Helium might be helping us some. However the benefits are probably not worth someone switching over from N2 to He unless they too have a "leaky" SEM.
Warren Straszheim
At 03:32 AM 8/2/98 -0600, Allen R. Sampson wrote: } No offense intended, but this discussion is getting way too anal. } I'm not sure what 'scattering' effects you believe helium will } improve. If you are referring to electron scattering during } operation, helium is probably harder for normal vacuum systems to } pump out then nitrogen, so the gain you are seeking more than likely } isn't there. I know of no practical difference between venting with } any dry gas. } } Conductive heat dissipation is always much faster than } convective. In the two or three seconds it takes you switch the } accelerating voltage off and to hit the vent button, the majority of } the heat built up in the small mass of a tungsten filament will } probably be conducted off through the filament posts and electrical } contacts in the gun. As the backfill gas works its way into the } gun, the remainder of the heat will be removed rather slowly since } the leak provided by a normal vent valve is rather small, taking 30 } seconds or more to completely vent. } } I have never seen or heard of anything other than very loose } anecdotal evidence for increases in cathode life dependent on venting } methods. At the most, I'll accept customers turning the filament } drive down before turning off the accelerating voltage and venting } the instrument. Anything more than that is simply unnecessary. It } is far more important to be cognizant of small vacumm leaks, } particularily in areas that could bleed into the gun area. Air leaks } in the gun during operation are extremely damaging. } } } At 02:52 PM 7/31/98 +1000, you wrote: } } } Dry oxygen maybe a lesser consideration, however, when the filament } } } chamber is to be vented, it's even more important to wait until the } } } filament has cooled. Exposure of a hot filament or LaB6 cathode to } } } N2 is not a good idea, exposure to O2 is worse. Cheers Jim Darley } } } ProSciTech Microscopy PLUS } } } } Hmmm. We vent our Hitachi with W filament with He (because of less } } scattering effect compared to N2). Should we be concerned about any } } reaction between the hot filament and the He? I would presume none. } } How about thermal shock effects? Anybody care to comment? } Allen R. Sampson } Advanced Research Systems } 317 North 4th. Street } St. Charles, IL 60174 } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net } WWW: http://www.mcs.net/~ars } Analytical instrument maintenance services }
} } Put a clean 0.22 micrometer filter on the syringe before use. } } Wet the filter with the solution before forcing a drop out } } which is to be used for a grid.
We have found that using a filter is not necessary. If the solution has been made-up and stored properly it does not require filtration.
} Hildy: I started dispensing lead citrate from a syringe recently, and find } it works pretty well. However, would you clarify/elaborate the part 'Wet } the filter...' ?
As I reported earlier, we store the made-up Reynolds in approx. 3 ml aliquots in Eppendorff tubes. We dispense by taking up the stain into a new disposable Pasteur pipette from about 4 - 5mm above the bottom of the tube and then discard the first drop. This minimizes the possibility that any contamination from crystals, dirt, etc, which may have accumulated at the bottom of the tube, on the surface of the stain or in the pipette will end up in the staining drop. Once the Eppendorff tube has been opened it is not re-used - it is discared with any left-over stain.
This uncomplicated method has produced almost totally contamination-free stain for many years, even in the hands of the most novice workers.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
Rick Felten wrote on 07/31/98 11:39 AM: } I would like to determine the thickness of Au and Ag on a copper substrate. } The copper is hundreds of microns thick; the Ag is a few thousand angstroms } and the Au is a few hundred angstroms. When the thickness of Ag is } required a Ag on Copper sample could be submitted (no Au). We have the } ability to make up varying (but unknown) thickness of Ag and Au and could } try to plate and cross section at a high angle or digest the deposit and } analyze with AA to get thickness standards. Or is it better to use pure Au } and pure Ag standards and let theory and computer software do the rest. } } Thanks } Ric
Hi Ric,
I once did some measurements of layer thickness of Au on Ho by the Bishop-Poole method (H. E. Bishop and D. M. Poole, J. Phys. D: Appl. Phys., vol. 6 (1973) 1142) with the use of a pure Au standard. The results were in good agreement with the thicknesses expected from the deposition parameters, so I think you can safely use pure standards and "let theory and computer software do the rest".
Best regards, Jorgen.
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
My old Link AN10000 system has a software package called TFOS (thin films on substrate) which does this calculation. I haven't used it for years but I seem to remember that it worked OK.
Regards
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
} } Rick Felten wrote on 07/31/98 11:39 AM: } } I would like to determine the thickness of Au and Ag on a } copper substrate. } } The copper is hundreds of microns thick; the Ag is a few } thousand angstroms } } and the Au is a few hundred angstroms. When the thickness of Ag is } } required a Ag on Copper sample could be submitted (no Au). We have the } } ability to make up varying (but unknown) thickness of Ag and Au } and could } } try to plate and cross section at a high angle or digest the deposit and } } analyze with AA to get thickness standards. Or is it better to } use pure Au } } and pure Ag standards and let theory and computer software do the rest. } } } } Thanks } } Ric }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
REGARDING in search for FEG-STEM in Europe
This message was forwarded to me by Bruce Davidson at = davidson-at-fisica.cib.na.cnr.it. Please respond directly to him.
In search of a cold-stage FEG-STEM in Europe
My name is Bruce Davidson from the Istituto di Cibernetica -CNR in = Naples, Italy. I'm searching for a FEG-STEM with cold-stage in Europe in = conjunction with a project to scribe Josephson junctions in YBCO thin films using a ~200 keV FEG source at ~150 K.
The idea for these junctions is to create a narrow region of oxygen displacement in a link patterned in a YBCO film by scanning a ~1 nm beam of electrons across the link once. The amount of oxygen = displacement determines the Tc reduction in the irradiated region, which cannot be = wider than a few nanometers in order to observe Josephson coupling across it. These junctions are some of the best-behaving, most uniform junctions = made to date in the high-temperature superconductors. We have fabricated many of these junctions in the past few years (the subject of my PhD thesis at = the Univ. of Wisconsin, August '97).
I am currently in Italy on a NSF-NATO Post-doc fellowship to expand the study of this technique. I would like to perform the irradiation of the = films using a STEM here in Europe rather than one in the US, if a suitable microscope can be found. The essential capabilities are:
1) FEG electron source (~0.5 nA current, ~1 nm FWHM spot -at- 200 keV). 2) scanning capabilities, with computer control of the scan coils. 3) sample cold stage, to maintain the sample at ~150 K during = irradiation.
Any information on suitable microscopes in Europe would be greatly appreciated.
A colleague is having problems getting good fixation of Xenopus neurons grown in culture, the membranes are shot by both SEM and TEM. She cannot add calcium to the fix due to certain experimental conditions. Any help would be greatly appreciated.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
If you have a diffusion pump or turbopumped SEM, the pump speed for different gas species goes as the molecular weight of the molecule. As a result, the pump speed for He compared to N2 are much lower. I could look it up, but I think that it is about an order of magnitude different. Another point is that, because the sensitivity of ion gauges is about 0.25 for He, the gauge pressure and the true pressure will be different. With a sensitivity of 0.25 for an ion gauge, the true pressure is four times the value displayed. I'm not sure what the values are for Penning gauges, but they are probably in the same ball park.
If your vacuum system has ion pumps, DO NOT backfill with an inert gas, especially He! Even with the special ion pumps that bury the inert gases into the plates, they still "burp" the inert gas. Helium pump speeds are extremely low.
One interesting aspect about your use of He around a hot filament is that it does have a higher thermal conductivity than N2, so I guess that it would cool the filament faster.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I should have mentioned that our Hitachi is a 2460N low vacuum SEM. During operation we He instead of air or N2 in order to reduce scattering effects at the 40 Pa of atmosphere that we maintain in order to cancel charging on our specimens. There was no intent of trying to reduce scatter by residual gas at 10-5 torr levels since He might have displaced heavier molecules.
I was intrigued that a side benefit of our He use might be increased filament life since the hot filament would not be exposed to N2 with its detrimental effects. A coworker likes to wait until the filament cools for a bit before venting the chamber (the gun chamber always gets vented with the specimen chamber). However, I had been supposing that thermal issues might be insignificant as you appear to corroborate. Since we just had a W filament go 441 hours even with sample changes about every 2 hours, I suppose that the Helium might be helping us some. However the benefits are probably not worth someone switching over from N2 to He unless they too have a "leaky" SEM.
Warren Straszheim
At 03:32 AM 8/2/98 -0600, Allen R. Sampson wrote: } No offense intended, but this discussion is getting way too anal. } I'm not sure what 'scattering' effects you believe helium will } improve. If you are referring to electron scattering during } operation, helium is probably harder for normal vacuum systems to } pump out then nitrogen, so the gain you are seeking more than likely } isn't there. I know of no practical difference between venting with } any dry gas. } } Conductive heat dissipation is always much faster than } convective. In the two or three seconds it takes you switch the } accelerating voltage off and to hit the vent button, the majority of } the heat built up in the small mass of a tungsten filament will } probably be conducted off through the filament posts and electrical } contacts in the gun. As the backfill gas works its way into the } gun, the remainder of the heat will be removed rather slowly since } the leak provided by a normal vent valve is rather small, taking 30 } seconds or more to completely vent. } } I have never seen or heard of anything other than very loose } anecdotal evidence for increases in cathode life dependent on venting } methods. At the most, I'll accept customers turning the filament } drive down before turning off the accelerating voltage and venting } the instrument. Anything more than that is simply unnecessary. It } is far more important to be cognizant of small vacumm leaks, } particularily in areas that could bleed into the gun area. Air leaks } in the gun during operation are extremely damaging. } } } At 02:52 PM 7/31/98 +1000, you wrote: } } } Dry oxygen maybe a lesser consideration, however, when the filament } } } chamber is to be vented, it's even more important to wait until the } } } filament has cooled. Exposure of a hot filament or LaB6 cathode to } } } N2 is not a good idea, exposure to O2 is worse. Cheers Jim Darley } } } ProSciTech Microscopy PLUS } } } } Hmmm. We vent our Hitachi with W filament with He (because of less } } scattering effect compared to N2). Should we be concerned about any } } reaction between the hot filament and the He? I would presume none. } } How about thermal shock effects? Anybody care to comment? } Allen R. Sampson } Advanced Research Systems } 317 North 4th. Street } St. Charles, IL 60174 } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net } WWW: http://www.mcs.net/~ars } Analytical instrument maintenance services }
For US citizens going to Mexico, you can use a passport, an out of date passport, birth certificate, or a certified copy of your birth certificate. -Scott ----------
I am not going to Cancun, but rather wish I was. However, I was just asked by a friend who has a son leaving for Mexico, what is needed by way of proof of identity for travel to Mexico? I know that neither passport or visa is necessary for travel to Canada. Is the same true of our southern neighbor?
Thanks to all who replied about necessary travel documentation. It looks like passports are still not officially required but would be advisable. I know I got a few mean looks coming back from Canada without one a couple years ago. As long as it doesn't get any worse than the mean looks, my friend should be fine.
The reason you are getting breakage is due to the shrinkage that processing and CPD in particular causes--up to 50% in some tissues. Since these are adherent cells, those neurites which are firmly adhered will probably show some form of breakage/damage. The only improvements I have obtained (and not consistently, I admit) has been using products such as Peldri or one of the hexamethyldisilazane products. This method is less consistent than CPD, but can yield better preservation of morphology of fragile samples. Contact any of the EM supply houses (EMS, SPI, Pella, Energy Beam Science, Fullam, Ladd, Polysciences) and ask for their technical fact sheets. There is also a fair amount of published methods literature--primarily from the 1980s. Hope this helps.
Roger Moretz, Ph.D. Toxicology
The comments are solely the opinions of the author. I have no vested interest in any of the products or the suppliers.
} -----Original Message----- } From: Sandy Perkins [SMTP:skperkin-at-vt.edu] } Sent: Friday, July 31, 1998 10:08 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM-neuroblastoma cells } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Hi- } } We are trying to refine our procedure for the preparation of } differentiated } neuroblastoma cells for SEM. The cells are grown on coverslips and we } have } used the following processing schedule: } } gentle phosphate buffer wash (0.1 M sodium phosphate, pH 7.4) } primary fixation in 2.5% glut/1% paraformaldehyde-1 hr } buffer washes } post-fixation in 1% osmium tetroxide-1 hr } ethanol dehydration } CPD } } The cells look OK, but a number of the neurite extensions are broken. } Has } anyone had any experience processing these cells for SEM? Do we need } a } "gentler" processing method? Any help would be greatly appreciated. } Thank } you very much. } } Sandy Perkins } } Laboratory for Neurotoxicity Studies } VA Tech }
I would be curious if anyone has any real data on filament life as affected by a cooling off period before chamber venting. Like many, I was trained to always cool a filament for several minutes before admitting air, but if it's really not necessary I'd be happy to stop. Sure would speed specimen turn-around times when we're running a bunch through.
Another question: do you really find it necessary to use 40 pa to eliminate charging on your 2460? Using both a 2460 and a 3200, I have almost always been able to eliminate charging with pressures of 1-5 pa, which should give substantially less scattering than 40 pa. Just wondering.
Finally---filament life of more than 400 hours is amazing!! And under variable pressure with frequent specimen changes! I'd love to know the secret to that trick. Maybe He is the wonder gas we've all been waiting for....
Regards,
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Hi: I operate a sem-edxra system, metallurgical microscope, LM and use Image Pro + and supporting equipment for Hallmark Cards Inc. (we also have an FTIR microscope). Over the years (20) I have been asked to do many unusual projects . One ongoing type of project I haven't been able to consistently do is to cross-section printed paper and measure the clay /clear top coat coating thickness' and determine the ink penetration of various inks. I have been able to use metallurgical procedures to obtain the clay coating thickness' but LR white hard mounting media makes most of the inks run plus the polishing takes 2 to 4 days (old equipment and user). I have talked to a couple of users of and one supplier of microtomes and they seem to think I need one (surprise) . I don't know anything about microtomes what type (rotary / sliding) and what blades or what mounting media would be most useful for us. There seem to be only one supplier of microtomes (Leica) are there any others? Does anyone have any recommendations or experience with this type of materials? Of course I will have to come up with justifications to buy one and any help would be appreciated. Thanks Terry Ellis email-tellis2-at-hallmark.com 816-545-6573 mail drop 359 2501 McGee, box 419580 K.C. MO 64141-6580
Randy writes ... } } } I would be curious if anyone has any real data on filament } life as affected by a cooling off period before chamber venting. } Like many, I was trained to always cool a filament ...
I'm not sure "real data" is needed. Tungsten's affinity for oxidation at elevated temperatures is well known. I believe most of us who don't wait for a cool down period are venting with dry nitrogen or some other inert gas.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Hi: I operate a sem-edxra system, metallurgical microscope, LM and use Image Pro + and supporting equipment for Hallmark Cards Inc. (we also have an FTIR microscope). Over the years (20) I have been asked to do many unusual projects . One ongoing type of project I haven't been able to consistently do is to cross-section printed paper and measure the clay /clear top coat coating thickness' and determine the ink penetration of various inks. I have been able to use metallurgical procedures to obtain the clay coating thickness' but LR white hard mounting media makes most of the inks run plus the polishing takes 2 to 4 days (old equipment and user). I have talked to a couple of users of and one supplier of microtomes and they seem to think I need one (surprise) . I don't know anything about microtomes what type (rotary / sliding) and what blades or what mounting media would be most useful for us. There seem to be only one supplier of microtomes (Leica) are there any others? Does anyone have any recommendations or experience with this type of materials? Of course I will have to come up with justifications to buy one and any help would be appreciated. Thanks Terry Ellis email-tellis2-at-hallmark.com 816-545-6573 mail drop 359 2501 McGee, box 419580 K.C. MO 64141-6580
Hi: I operate a sem-edxra system, metallurgical microscope, LM and use Image Pro + and supporting equipment for Hallmark Cards Inc. (we also have an FTIR microscope). Over the years (20) I have been asked to do many unusual projects . One ongoing type of project I haven't been able to consistently do is to cross-section printed paper and measure the clay /clear top coat coating thickness' and determine the ink penetration of various inks. I have been able to use metallurgical procedures to obtain the clay coating thickness' but LR white hard mounting media makes most of the inks run plus the polishing takes 2 to 4 days (old equipment and user). I have talked to a couple of users of and one supplier of microtomes and they seem to think I need one (surprise) . I don't know anything about microtomes what type (rotary / sliding) and what blades or what mounting media would be most useful for us. There seem to be only one supplier of microtomes (Leica) are there any others? Does anyone have any recommendations or experience with this type of materials? Of course I will have to come up with justifications to buy one and any help would be appreciated. Thanks Terry Ellis email-tellis2-at-hallmark.com 816-545-6573 mail drop 359 2501 McGee, box 419580 K.C. MO 64141-6580
My apologies for not being more specific in my first post. The question should have been "Does anyone have a recommendation for a good TEM/SEM fixative for Xenopus neurons grown in culture?" My colleague has had very poor TEM results with 2.5% glut in 0.1M cacodylate with 0.1M sucrose, there are no membranes visible. Cell membranes, mitochondrial membranes, organelle membranes, all gone/invisible. Yes, post-fixation with osmium was used. Overall ultrastructure is also poor. She has also had very poor SEM results with the same fix with the addition of 4% paraformaldehyed, plasma membrane had large holes. When I suggested adding calcium to the fix I was told that the experimental protocol prohibited this. Any help will be appreciated!
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
At 11:54 AM 8/3/98 -0600, Randy Tindall wrote: } I would be curious if anyone has any real data on filament life as affected } by a cooling off period before chamber venting. Like many, I was trained } to always cool a filament for several minutes before admitting air, but if } it's really not necessary I'd be happy to stop. Sure would speed specimen } turn-around times when we're running a bunch through.
One operator likes to cool the filament for a while to get longer life. But this filament has had a number of quick cool downs anyway. If it is a reaction with nitrogen, then maybe we can vent the He to it right away. Then again, we ran the scope in automatic mode with a beam shut off at the end of the run. It might have had plenty of time to cool for most exchanges.
} Another question: do you really find it necessary to use 40 pa to eliminate } charging on your 2460? Using both a 2460 and a 3200, I have almost always } been able to eliminate charging with pressures of 1-5 pa, which should give } substantially less scattering than 40 pa. Just wondering.
We have tried reducing the pressure and have run into charging. But we run a lot of beam current to our concrete samples, around 1-2 nA by my guess. Maybe the lower pressures work for lower beam current.
} Finally---filament life of more than 400 hours is amazing!! And under } variable pressure with frequent specimen changes! I'd love to know the } secret to that trick. Maybe He is the wonder gas we've all been waiting } for....
I just hope that we can replicate the performance. Our normal life has been in the 100 hour range.
I have been asked to organize a session on precision specimen prep for the 1999 MSA/MAS meeting in Portland. Here is the descriptive paragraph for the Call for Abstracts:
======
Precision Specimen Preparation
In the physical sciences we see an increasing need to prepare TEM specimens of very small pre-selected locations in a variety of different types of samples. The main driving force for high spatial resolution specimen preparation is the semiconductor industry, where the size of the target locations to be prepared have diminished to a point where they cannot be viewed in a visible light microscope. Precision preparation of the impact point of a projectile into armor or an analysis of the precise initiation point of a micro-crack are other examples. This symposium seeks to review the state of the art of precision specimen preparation and to explore the prospects for improved spatial resolution in the near future.
======
As I've been correctly accused of having a bias towards tripod polishing, I am looking for suggestions (self-suggestions are fine!) for people to ask to be Invited Speakers in this symposium that would bring a broader perspective to the topic. I would be especially interested in someone to address non-semiconductor examples. I could use a symposium co-chair too, if there is one out there.
Now that MSA's middle school microscopy manual is published, Project MICRO (Microscopy In Curriculum - Research Outreach) is going into high gear. Several MSA local societies already have outreach programs, or are about to begin one (Arizona, Minnesota, North Carolina, Oklahoma,). So Nestor and I plan to expand the MICRO web page (yes, that means that the wonderful microscopy quotes that you folks sent last spring will be posted soon). Many of you have been visiting schools for a long time, and you've developed good materiel. One resource that we want to offer is a collection of successful classroom-tested exercises that can be used to extend the experiences provided by "Microscopic Explorations". Grade level? Elementary & middle school. Topic? Anything that works well. Reward? Our thanks, and the opportunity to share the results of your hard work. Please respond directly, since your text may be lengthy; I'll organize and post.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
As you all know the cathode assembly gets up to a pretty high temperature=
too! If we vent with "dirty" air we cause contamination to deposit on th= e hot cathode. I agree the filament cools very quickly but we all know the=
cathode does not?
Waiting a few minutes after getting the READY indication before switching=
on the filament and a few minutes after switching off the filament, prior=
to letting in AIR, the result a difference in filament life and cathode contamination. =
The first point is that the READY indication means the vacuum is only jus= t good enough to use. Wait a few more minutes and the improvement in vacuu= m will help give a better filament life and keep the column clean longer. =
Put a beam down a column with a poor vacuum and you put up the contamination! From the other direction allowing the cathode to cool pri= or to letting in air will keep this cleaner too!
Just the same when you clean a column, it makes sense to clean the column=
on a Friday afternoon, pump and wait over the weekend before putting a beam down the column. Clean a column and then pass a beam through it means that most of the vapours from the cleaning media have just deposite= d on your clean components!
Steve Chapman Senior Consultant E.M. Protrain for courses and consultancy in electron microscopy world wide Tel & Fax 44 (0)1844 353161 WWW http://ourworld.compuserve.com/homepages/protrain
Terry Ellis wrote: =============================================== ..........One ongoing type of project I haven't been able to consistently do is to cross-section printed paper and measure the clay /clear top coat coating thickness' and determine the ink penetration of various inks. I have been able to use metallurgical procedures to obtain the clay coating thickness' but LR white hard mounting media makes most of the inks run plus the polishing takes 2 to 4 days ...... ================================================ We have been doing this kind of sample for some number of years and the microtome manufactures are correct, you really can only do this kind of study properly via the use of an ultramicrotome (and diamond knives, glass would be a waste of time). Because the acrylic binders, both in the ink and the paper substrate, have low Tg's, the sectioning in general has to be done cryo. The views of the structure by examining the sections by TEM are so far superior to the best we have been able to obtain by SEM that now we rarely even make the SEM attempt. Questions of ink and pigment penetration you can not come close to in an SEM anyhow.
There is a "trick" that you have to employ to keep the ink from literally dissolving in (or being swollen by) the embedding resin and that is to sputter coat a layer of platinum (we prefer Pt for this purpose over Au because it is less likely to smear during sectioning) onto the surface of interest and THEN do the embedding. Since paper has some amount of porosity , to be on the safe side, we always do a vacuum embedding. If the paper has been in a humid environment, it might be wise to just "pump on it" in a vacuum evaporator over night to pull of some of that unwanted moisture.
We have consistently gotten our best results with our own SPI-Pon™ 812 resin system (but I suspect that at least some of the "Epon replacements" from others would work equally well). With a little bit of practice, you can get the resin to cure in a way that there is virtually no interaction with the sample underneath the Pt (or Au) passivation layer. This layer has the added benefit of making very clear that you are seeing the entire ink layer cross-section and not seeing an artifact because part of it broke away during the sectioning.
The other point has to do with the selection of the diamond knife. There are two considerations, and this part of my posting might be "controversial" : a) Use only "materials science" diamond knives for this kind of work, you will save a lot of money if you do that, and b) follow our (on the website) advice about knife angle, use a 45 deg. angle and not the 55 deg. angle sometimes touted as being "right" for "materials science" work. At least for these kinds of samples, if you have any kind of serious compression effects present, the layers will quite readily split apart, so you want to minimize compression effects by using a 45 deg. angle knife. Will the "sharper" knife edge "wear out faster"? Probably, but not as much as you might think since when compared to a 55 deg. knife, remember that in order to get good sections, you have to make many more sections just to get to that point. And that puts all kinds of additional wear and tear on the knife edge! So the economics are not exactly what common sense might suggest!
The last point has to do with the data presentation. Again, it has been our experience that the deposition of ink is inherently not necessarily all that homogeneous of a process and therefore, we take the micrographs as a montage along the surface (ink) layers. The objective is to show the entire "forest" and not just individual trees. It is the appearance of the forest that more often than not, shows the differences between samples that behave differently. If you are not doing these kinds of samples as montages, you could very easily "miss" what would otherwise be there to be seen.
Disclaimer: SPI Supplies offers the embedding resins and diamond knives to do this kind of work and if this all sounds too complicated to gear up to do it yourself, the laboratory services part of our company would be happy to talk to you about doing it for you.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
In a message dated 98-08-03 15:36:02 EDT, tellis2-at-hallmark.com writes:
{ { One ongoing type of project I haven't been able to consistently do is to cross-section printed paper and measure the clay /clear top coat coating thickness' and determine the ink penetration of various inks. I have been able to use metallurgical procedures to obtain the clay coating thickness' but LR white hard mounting media makes most of the inks run plus the polishing takes 2 to 4 days (old equipment and user). I have talked to a couple of users of and one supplier of microtomes and they seem to think I need one (surprise) . I don't know anything about microtomes what type (rotary / sliding) and what blades or what mounting media would be most useful for us. There seem to be only one supplier of microtomes (Leica) } }
Hi Terry,
I'll sidestep the issue of manufacturers of microtomes, since I'm sure you will hear from all of us who manufacture and sell them. And I like to avoid flames from the SysOp whenever possible...
Actually, I used to do some similar work on paper products in one of my previous lives, and I can enthusiastically recommend cryomicrotomy as the way to go. You can freeze the specimen directly in one of several aqueous or non- aqueous embedding media and section it with either glass, diamond or tungsten carbide knives.
I would recommend a rotary microtome with a cryosectioning kit. The cryo kit is a box that is cooled with liquid nitrogen and has temperature regulation and heaters built in for both the specimen and the knife. The cryobox fits on the microtome in place of the regular knife holder.
If you have a cryostage on the SEM you can transfer the sectioned ("polished") material directly to the scope for evaluation. If not, you can melt the sectioned material out of the embedding media, rinse quickly, dry, then mount and examine in the SEM.
Of course you can also collect the sections and mount them for FTIR.
The main advantages of cryotechniques are bypassing all of the solvents, plastics, abrasives/polishing, etc. that are normally used in specimen prep, and the quick turnaround time. From start to finish, it takes maybe 10 minutes after the cryosectioning device is cooled down.
And yes, Leica makes such equipment. So do a couple of others. I'm sure they will also respond.
Best regards, Bob ***************************************** Robert (Bob) Chiovetti rchiovetti-at-aol.com E. Licht Company / 1-800-865-4248 Colorado/Utah/Wyoming/Arizona/ New Mexico/West Texas U.S.A. Representing Leica Since 1967 *****************************************
Forgive me, Nestor, if this violates your directive about Cancun.
But there have been some rather major fare reductions to Cancun and if you call your airline, you might quality for a nice refund of the difference. That has just saved the two of us a cool $140 on two tickets. At least it worked that way on United.
I expect that filament life has nothing to do with the use of any inert gas. In the 1950th (no, I was not in labs then), a paper was published The life of filaments. My long-term memory recall advises that short of physical violence, filament life is determined by vacuum, if it's under 5x10-5 torr and by emission if it's above 15microamps. It is assumed that throughout it's life the filament is correctly saturated. I recall that there was a curve (or maybe just a number of data points) showing filament "death" at various vacua and emissions. In modern instruments, emission is most frequently the limiting factor. Reaching a filament life of 500 hours is no trouble in microprobes at say 10 to 15 microamps, it's a little less for a TEM, which commonly run at 30 microamps. Tell me when a SEM operating at 80 or more microamps reaches 500 hours filament time; then it will be time to rewrite that 1950ish paper. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Tuesday, 4 August 1998 3:55, Randy Tindall [SMTP:rtindell-at-NMSU.Edu] wrote: . . . . . . } Another question: do you really find it necessary to use } 40 pa to eliminate } charging on your 2460? Using both a 2460 and a 3200, I } have almost always } been able to eliminate charging with pressures of 1-5 pa, } which should give } substantially less scattering than 40 pa. Just wondering. } } } Finally---filament life of more than 400 hours is } amazing!! And under } variable pressure with frequent specimen changes! I'd } love to know the } secret to that trick. Maybe He is the wonder gas we've } all been waiting } for.... } } Regards, } } Randy } } } Randy Tindall } Electron Microscope Laboratory } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } } rtindell-at-nmsu (work) } nrtindall-at-zianet.com (home)
your travel agent or the airline will give you a tourist card (actually a flimsy set of thin papers). There is no charge. Part is collected or stamped when you enter, and the remainder is surrended when you leave. The other docs are useful.
I have been noticing a black precipitate on my grids. At high magnification, the precipitate looks like large hexagon chrystals. When the electron beam hits the precipitate, it tears the tissue. Any good hints as to what could be my problem would be of great help.
Moreover, would you happen to know of any jobsites on the net devoted to microscopy?
Thank you for your time,
Maria L. Sunio
P.S. Please email me directly to sunio-at-utsw.swmed.edu
This message is in MIME format. The first part should be readable text, while the remaining parts are likely unreadable without MIME-aware tools. Send mail to mime-at-docserver.cac.washington.edu for more info.
We have great pleasure to inform you about our conference "COLOR '99" organised in Krakow under the auspices of Foundry Research Institute, Krakow, Poland.
Below we send to you the first announcement of this conference in the file Word 7.0 "First.doc".
Best regards.
Members of Local Conference Committee:
Janina Radzikowska and Krzysztof Jan Huebner
================================================
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
} [skip all the stuff I agree with] Clean a column and then pass a beam } through it means that most of the vapours from the cleaning media have } just deposited on your clean components! } Even worse. The radiolysis products--ions, free radicals, etc.-- will deposit on and possibly interact with the column. Yours, Bill Tivol
We had success using 1% glutaraldehyde/0.25% acrolein in 10mM MgCl2, 50mM PIPES, pH 6.5. See Luther et al, J. Neurocytol 25, 417-427, 1996.
Steve Samuelsson ______________________________ Reply Separator _________________________________
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Dear List:
A colleague is having problems getting good fixation of Xenopus neurons grown in culture, the membranes are shot by both SEM and TEM. She cannot add calcium to the fix due to certain experimental conditions. Any help would be greatly appreciated.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
by imo28.mx.aol.com (IMOv14_b1.1) id NGPa017153 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 4 Aug 1998 10:57:22 -0400 (EDT) Message-ID: {bc26a28c.35c72153-at-aol.com}
Dear all, I am placing this message for a third party, looking for a short course in electron microscopy preferably in regard to clinical pathology.
Unfortunately I missed prvious listings. Any information is wellcome.
Eckhart C. Dorneich Leo Electron Microscopy, Inc. ECDorneich-at-aol.com
Leica CLSM - confocal microscope (inverted). The system includes: a Leitz Fluovert inverted microscope, objective lenses, filters, krypton/argon laser, Motorola computer (running OS9) with Leica ScanWare software, and 3 monitors. Also for sale is a Focus Graphics image recorder.
If interested, please contact:
Scott Henderson, Ph.D. Director of Microscopy, Dept. Cell Biology & Anatomy, Mount Sinai School of Medicine, Box 1007, 1 Gustave L. Levy Pl., New York, NY 10029-6574
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } About staining with Reynolds lead citrate } } } } Put a clean 0.22 micrometer filter on the syringe before use. } } } Wet the filter with the solution before forcing a drop out } } } which is to be used for a grid. } } We have found that using a filter is not necessary. If the solution } has been made-up and stored properly it does not require filtration. } } } Hildy: I started dispensing lead citrate from a syringe recently, and find } } it works pretty well. However, would you clarify/elaborate the part 'Wet } } the filter...' ? } } As I reported earlier, we store the made-up Reynolds in approx. 3 ml } aliquots in Eppendorff tubes. We dispense by taking up the stain into } a new disposable Pasteur pipette from about 4 - 5mm above the bottom } of the tube and then discard the first drop. This minimizes the } possibility that any contamination from crystals, dirt, etc, which } may have accumulated at the bottom of the tube, on the surface of the } stain or in the pipette will end up in the staining drop. Once the } Eppendorff tube has been opened it is not re-used - it is discared } with any left-over stain. } } This uncomplicated method has produced almost totally } contamination-free stain for many years, even in the hands of the } most novice workers. } } Robin H Cross } Director : EM Unit, Rhodes University, Grahamstown, South Africa } eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 } http://www.ru.ac.za/affiliates/emu/em.htm } Sounds good to me! You have worked out a good method of making the stain which is of critical importance in the avoidance of precipitates. (By "wet the filter" I mean to drive a few drops of lead through the filter and discard these drops). If your method works, do not change it. I have heard of others who use Eppendorfs for storage successfully. (I am basically lazy (call it practical) and when I make up stain I make huge amounts like 500ml, and I distrubute that into 10 syringes, and I don't have to worry about it for 2 years). P.S. I hate making up stain. Bye, Hildy
NCEM is a national user facility housing state of the art electron microscopes and advanced image analysis and specimen preparation facilities, dedicated to the electron-optical microcharacterization of materials. The Center currently has an opening for a one year term Visiting Postdoctoral Scientist to: - Conduct research on Microstructural characterization of plastically deformed samples using the wide range of TEM facilities available at NCEM. Explore the correlation between local physical/chemical microstructure in plastically deformed regions with local changes in magnetic structure (measured independently by SQUID imaging techniques. - Develop Lorentz imaging methods on NCEM's CM200FEG microscope and appropriate computer modeling methods to interpret Lorentz images.
Qualifications: Ph.D.(within the last four years) in Materials Science/Physics or related area. Very strong hands-on experience in various TEM techniques and their application to microstructural characterization is required. Experience in Lorentz Imaging and TEM specimen preparation is desirable. Computing skills, particularly in developing/adapting code for micromagnetic modeling will also be helpful.
Send resume and cover letter to:
Dr. Kannan Krishnan, Lawrence Berkeley National Laboratory, One Cyclotron Road, M/S-72, Berkeley, CA 94720. Phone: 510/486-4614, Email: Krishnan-at-lbl.gov Fax: 510/486-5888
Good day everone. Does anyone know a current source for Janssen immunogold reagents? Thanks in advance. Robert Cox Shriners Hospital Burns Institute Galveston, Texas
Good day everone. Does anyone know a current source for Janssen immunogold reagents? Thanks in advance. Robert Cox Shriners Hospital Burns Institute Galveston, Texas
I am assuming the cells are fixed as a monolayer. I have sometimes had trouble with the osmolarity requirements of cell cultures. Check that. If they are a monolayer I would start with a more dilute fixative, such as 1 to 1.5 percent glut to help keep the osmolarity down, and you are only working with a few cells thick. We also add (10 percent by vol. ) saturated picric acid to stabilize membranes during aldehyde fixation. One investigator I worked with added 0.05M Potassium ferricyanide with the osmium to enhance neural tissue.(ref ?) If your fixative is refrigerated bring it to room temp so as not to shock the cells. Good luck with your cells.
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Date 8/4/98 Time: 6:08 PM Internal Memorandum
Does a commercial Thermal Wave Imaging Microscopy facility exist in the San Francisco Bay Area? If so please email me at Hwachob-at-exponent.com. Thanks you for your assistance.
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Does a commercial Thermal Wave Imaging Microscopy facility exist in the San Francisco Bay Area? If so please email me at Hwachob-at-exponent.com. Thanks you for your assistance.
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} My colleague has had very poor TEM results with 2.5% glut in 0.1M } cacodylate with 0.1M sucrose, there are no membranes visible. Cell } membranes, mitochondrial membranes, organelle membranes, all } gone/invisible. Yes, post-fixation with osmium was used.
I suspect that the dehydration steps were too prolonged. For a thin layer of cells 3-5 minutes in each step should be plenty, and not too many steps ... e.g. 50% ethanol, 70%, 90%, 2 x absolute, propylene oxide, then resin.
} She has also had very poor SEM results with the same fix with the } addition of 4% paraformaldehyed, plasma membrane had large holes.
Sorry I can't offer advice on this, except to suggest using fixative at culturing temp (? 37 degC) and not ice-cold, and ensuring that the pressure changes in the CPD are not too sudden (both when first adding CO2 and at the end when returning to atmospheric pressure).
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
At 14:32 04/08/98 +0200, Krzysztof Jan Huebner wrote: } } } } Dear Friends, } } We have great pleasure to inform you about our conference "COLOR '99" } organised in Krakow under the auspices of Foundry Research Institute, } Krakow, Poland. } } Below we send to you the first announcement of this conference in the } file Word 7.0 "First.doc". } } } Best regards. } } } Members of Local Conference Committee: } } Janina Radzikowska and Krzysztof Jan Huebner } } } ================================================ } } } Krzysztof Jan Huebner }
I'm not interested by your conference but I received your attached file and it could be too much waste time for many people of the list server if everybody use this way each time there is a conference in the world. Please next time send only E.mail. Best regards Nicolas Stephant
We have great pleasure to inform you via e-mail about our conference "COLOR '99" organised in Krakow under the auspices of Foundry Research=20 Institute, Krakow, Poland.
Below we send to you the first announcement of this conference.
International Conference on Color Metallography Krak=F3w, Poland 19 - 21 May 1999
under the auspices of Foundry Research Institute Krak=F3w, Poland
Scope: This conference will be a meeting of scientists and industry workers interested in application of metallographic color techniques to=20 microstructural investigations of metals and nonmetallic materials with the use of the light microscope. The techniques of specimen preparation=20 will be presented and discussed as well. Exhibition of=20 instruments, equipment and photographic materials will make it possible=20 for the participants to get acquainted with the latest achievements in=20 the research and investigation technology. Conference'will be held in the= =20 MANGHHA Centre of Japanese Art and Technology in Krak=F3w with a beautiful= =20 view to Wawel castle.
Scientific Committee: G.F. Vander Voort (USA) - Chairman G. Petzow (Germany) P Skocowsky (Slovakia) K. Huebner (Poland) J. Radzikowska (Poland)
Main Topics of the Conference: 1. Preparation of specimens for color metallography 2. Light optical methods for color imaging: polarized light dark field differential interference contrast =B7 fluorescence 3. Color etching methods: tint etching anodizing interference layer method gas contrasting/reactive sputtering methods 4. Color image analysis procedure 5. Application of color metallography 6. Documentation of color images 7. New color techniques
Program: The scientific program will consist of keynote lectures, oral and poster=20 presentation and metallographic contest of color microphotographs.
Abstract submission: Abstracts should be in English and contain from 50 to 200 words on A4=20 sheet with wide margins. Along with the abstract, please indicate the=20 technical topic for which the abstract is being submitted; the title of the= =20 proposed presentation; and complete author information (name, address, phon= e, fax, E-mail) for all authors, with the presenting author listed first.=20 Alternatively, you can send MS Word 7 file (with True type fonts included)= =20 as an attachment to E-mail.
Before submitting your abstract or any other form of work listed in the=20 program, please be informed that all costs associated with participation=20 in the Conference will be at your expense, including travel, housing and=20 conference registration. Abstracts should be submitted to Conference Secretariat.
Deadline: 15 October 1998 for abstracts 30 January 1999 for papers 30 April 1999 for posters and contest microphotographs
Requirements regarding papers, posters and microphotographs as well as information referring to costs of participation and final program will be= =20 distributed later in the second announcement.
Conference language: English
Local Organizing Committee: Krzysztof Huebner Janina Radzikowska Halina Pawlowska Janina Danczak Krystyna Luszczkiewicz
Conference Secretariat: Krzysztof Huebner Foundry Research Institute Zakopianska 73 30-418 Krak=F3w POLAND Fax: (48-12) 266 08 70 Phone: (48-12) 266 50 22 ext.356 or 316 E-mail: hubner-at-iod Krak=F3w.pl.
Signature:
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D Krzysztof Jan Huebner=20
{hubner-at-IOd.krakow.pl} :-)=20
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
by jess.gla.ac.uk with esmtp (Exim 2.01 #1) for Microscopy-at-sparc5.microscopy.com id 0z430H-0002lD-00; Wed, 5 Aug 1998 13:49:17 +0100 Received: from l-tetley.chem.gla.ac.uk (l-tetley.chem.gla.ac.uk [130.209.221.186]) by lenzie.cent.gla.ac.uk (8.8.4/8.8.8) with SMTP id NAA27148 for {Microscopy-at-Sparc5.Microscopy.Com} ; Wed, 5 Aug 1998 13:49:15 +0100 (BST) Message-Id: {3.0.1.16.19980805134914.402fa156-at-udcf.gla.ac.uk} X-Sender: gbza40-at-udcf.gla.ac.uk X-Mailer: Windows Eudora Light Version 3.0.1 (16)
FOOD MICROSTRUCTURE - Towards the Year 2000 and Beyond
This conference is being held in collaboration with the Royal Microscopical Society
An understanding of the structure of foods and food products is essential to the efficient development of new processes or products. The future of the food industry is closely related to new developments in food microscopy.
This conference is an opportunity for those involved in food product development to hear the latest findings from key scientists in the field of food microscopy. The meeting will act as a forum to establish the needs of the food industry over the next decade, to discuss recent advances in food microstructural research and to evaluate new microscopical techniques.
This conference is aimed at food microscopists, food technologists and research scientists, particularly those involved in the development of new products or processes. Any company engaged in product development should be represented at the conference, to ensure that it is aware of the opportunities that advances in food microscopy can offer.
The aim of this conference is to:-
bring together food microscopists and food technologists to enable fundamental and applied research to be presented together;
highlight recent advances in food microstructural research;
provide a forum for discussion of microstructural studies relating to food;
identify the potential of new techniques.
Conference organisers: Mrs Kathy Groves and Dr Morag Saunders, Leatherhead Food RA; Dr Ashley Wilson, CCTR University of York.
There is still time for other papers to be submitted - please contact the conference organisers for further information.
KEYNOTE PAPER: Structure - the Only Thing that Matters? - Professor Peter Lillford, CBE, Unilever Research The challenge facing members of the food industry is to become architects of structure rather than processors of materials. This will need better application and novel approaches to microscopy.
Invited Paper: Cryogenic Environmental SEM of Ice Cream - an Introduction to the Principles of ESEM with Applications to Observations on Ice Cream - Brad Theil, Polymers and Colloids, Physics Department, University of= Cambridge
Food Microstructure using X-ray Projection Microscopy - John Judge and Peter Lillford
LUNCH
Chairman: David F. Lewis, SAC
The X-ray Microscope and its Applications to the Food Industry - Simon Burgess, Oxford Instruments
Probing Food Biopolymer Functionality with Atomic Force Microscopy (AFM) - Vic Morris, Institute of Food Research, Norwich (co-authors: A.P. Gunning, A.R. Kirby, A.R. Round, A. Mackie and P. Wilde)
The Environmental Scanning Electron Microscope - Speaker from Philips Electron Optics
Observations of Food Microstructure by Environmental Scanning Electron Microscopy - Debbie Stokes, Polymers and Colloids, Physics Department, University of Cambridge (co-author: A.M. Donald)
SESSION 2: APPLICATIONS OF MICROSCOPY TECHNIQUES IN FOOD RESEARCH
Chairman: Ashley Wilson, CCTR University of York
F Invited Paper: Applications of Variable Vacuum in Food Microscopy - Roger Angold, RHM Technology
F FTIR Microscopy in Troubleshooting for New Product Development - Hilary Holgate, RSSL
F Using Image Analysis and Confocal Microscopy Combined to Measure Deformation in Starch Matrices - Jeremy Addler, RHM Technology
F The Microscopy of Food. ESEM and Confocal Microscopy - Complimentary Techniques - Anthony Robinson, Polymers and Colloids, Physics Department, University of Cambridge
F Invited Paper: The Use of Electron Microscopy in the Investigation of BSE - Michael Stack, Veterinary Laboratories Agency
LUNCH and opportunity to view trade exhibition
SESSION 3: MICROSCOPY OF FOOD PRODUCTS, PROCESSES AND INGREDIENTS
Chairman: Roger Angold, RHM
Invited Paper: Microscopy - the Art of Food Technology - Professor Anne-Marie Hermansson, The Swedish Institute for Food and Biotechnology
Changes in Endosperm Structure during the Production of Popped Grain - Mary Parker, Institute of Food Research, Norwich
Confocal Laser Scanning Microscopy of Dairy Products and Ingredients - Methodology and Some Applications - Mark Auty, Dairy Products Research Centre, Moorepark, Ireland
Poster Exhibition and Opportunity to view Trade Exhibition
SESSION 3 (continued): MICROSCOPY OF FOOD PRODUCTS, PROCESSES AND INGREDIENTS
Chairman: Professor Anne-Marie Hermansson, SIK
Invited Paper: Brian Brooker, Institute of Food Research
The Use of SEM, Confocal Microscopy and Instrumental and Sensory Techniques in the Study of Biscuit Texture - Michael Edwards, Campden and Chorleywood Food RA
Crystallising Carbohydrates - Use of the Microscope in Understanding Crystallising Structures in Heat-resistant Chocolate and Powdered Sorbitol - Kathy Groves, Leatherhead Food RA
F Microstructural Changes of Food Products Following Inclusion of Non-starch Polysaccharides and Fibre Supplements: Implications to Dietary Nutrition - Karen Linton, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth (co:authors: C. Brennan and P. Moura de Magallraes)
LUNCH
Chairman: Brian Brooker, IFR, Reading; Mrs Kathy Groves, Leatherhead Food RA
Ultrastructural and Textural Changes in Heat-processed Fruits - Morag Saunders, Leatherhead Food RA
Cereal Structure: its Relationship to Raw Material Quality and End-product Utilisation - Charles Brennan, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth (co-authors: K. Linton, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth; D. Griggs, Pauls Malt Ltd; I. Cantrell, Pauls Malt Ltd; P.R. Shewry, IACR - Long Ashton)
Invited Paper: Perspectives on Food and Microscopy - David Lewis, SAC
CLOSE=09
ADMINISTRATION
Registration and closing times will be advised with the acknowledgement of your booking=09
Customized, on-site courses are available through MME. We have a consultant who has been doing clinical EM for a number of years. Please see our web-site for further information:
Are there other SEM, TEM, AFM,STM, EDS,WDS, Materials Analysis and Characterization Listserers? JD
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
I would recommend you to look at http://www.mwrn.com/ I found it to be very imformative in the area of microscopical resources
A ---------- =CE=F2: charles j day =CE=F2=EF=F0=E0=E2=EB=E5=ED=EE: 6 =E0=E2=E3=F3=F1=F2=E0 1998 =E3. 10:33 =CA=EE=EC=F3: Microscopy-at-sparc5.microscopy.com =D2=E5=EC=E0: Other EM and Materials Analysis Listserers?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Are there other SEM, TEM, AFM,STM, EDS,WDS, Materials Analysis and Characterization Listserers? JD=20
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
If anyone has information or recommendations for embedding mud samples for TEM observation, I would appreciate it. To date, I have had some success with LR White resin in the microwave.
We are in need of a good atlas identifieng the ultrastructure of viruses. does anyone have any suggestions.
Christine Lee , Veterinary Pathology, University of Queensland Christine Lee, Veterinary Pathobiology, University of Queensland. C.lee-at-mailbox.uq.edu.au
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I'm looking for a method to stain tissue en bloc so that I can see it while I'm cryo-sectioning -- something with just a hint of color.
The tissue is frog saccule, which is small and so translucent that I can't see it as I approach it through the matrix, which is OCT. The tissue is destined for immunocytochemistry, so antigenicity needs to remain preserved. (It would be helpful to find a stain that may be used for other tissue types as well -- mouse retina and chick utricle, for example.)
I was told that methylene blue may be used, but I've been unable to find anything in my reference texts.
I would be grateful for any suggestions.
Thanks,
Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu
Center for the Study of the Biology of Hearing and Deafness Central Institute for the Deaf 818 S. Euclid Ave. St. Louis, MO 63110
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. I have a pair of glasses (polycarbonate lenses) which have become scratched. The opticians I have consulted say that there is nothing to be done except buy new lenses. The scratches are not deep. They are very fine but, since they are in the middle of the lenses, they render the glasses nearly useless.
Surely with all the expertise this community has in polishing materials of every kind, someone can tell me how to save $100.
Alwyn Eades . Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
This is a little off the topic, but microscopists who wear plastic lenses are bound to get them scratched. I have an answer for you, but it takes some elbow grease.
There are kits available for repairing CDs. Even though they may boldly say that they "fill" in scratches, the two that I have tried consisted of an abrasive solution and a soft pad. Make sure that you have removed all the dirt from your glasses and apply the opaque white liquid and rub with the pad. After a few minutes you should see the scratches "rub out."
I have even had moderate success restoring CDs!
Best regards from your token Optical Engineer mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
I know this has been covered in the distant past, but I never wrote it down. We have an open house coming up and I would like to have a micrograph from a CD for show and tell. Could someone tell me the sample prep steps to do this? I've never done it before.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Maybe you do require a printed atlas and somebody else will advise. However, there are many sites on the internet that feature virus and at least a couple with quite extensive collections. Go to our links page http://www.proscitech.com.au/links.htm and use the browsers control/F function to search for "virus" and "virol" Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Friday, 7 August 1998 2:44, Christine Lee [SMTP:C.Lee-at-mailbox.uq.edu.au] wrote: } ---------------------------------------------------------- } --------------
} We are in need of a good atlas identifieng the } ultrastructure of viruses. } does anyone have any suggestions. } } } Christine Lee , } Veterinary Pathology, } University of Queensland } Christine Lee, } Veterinary Pathobiology, } University of Queensland. } C.lee-at-mailbox.uq.edu.au }
I wonder if there is anyone working with EBIC who can provide me with = some advice. I have been asked to try and produce data from ZnO = varistors. In particular they want to relate the structure to current = flow across grain boundaries. I must point out that I am a complete novice to EBIC. I have obtained = a simple Specimen Current Monitor ( GW Electronics Type 31 ) which is = now connected to the microscope and producing EBIC images. The problem = is when I try to apply current biasing ( +/- 10V ), from an alkaline = battery, to a 5.5V device the only effect is banding/interference. = This may be due to a bad contact ( I will be making a new regulator ), = but there is no change in the EBIC image at the grain boundaries. I = have tried a range of KV's and probe currents without success. I would be very grateful if anyone can provide me with some advice.
Thanks,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} Hi, {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} I wonder if there is anyone working with EBIC who = can provide=20 me with some advice. I have been asked to try and produce = data from=20 ZnO varistors. In particular they want to relate the = structure to=20 current flow across grain boundaries. {/FONT} {/DIV} {DIV} {FONT size=3D2} I must point out that I am a complete novice to=20 EBIC. I have obtained a simple Specimen Current Monitor ( GW =
Electronics Type 31 ) which is now connected to the microscope and = producing=20 EBIC images. The problem is when I try to apply current = biasing (=20 +/- 10V ), from an alkaline battery, to a 5.5V device the only effect is =
banding/interference. This may be due to a bad contact ( I = will be=20 making a new regulator ), but there is no change in the EBIC image at = the grain=20 boundaries. I have tried a range of KV's and probe currents = without=20 success. {/FONT} {/DIV} {DIV} {FONT size=3D2} I would be very grateful if anyone can provide me = with some=20 advice. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Thanks, {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Colin {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Colin Reid, {BR} Electron Microscope=20 Unit, {BR} Trinity College Dublin, {BR} Dublin = 2, {BR} Ireland. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Tel: 353-1-6081820 {BR} Fax:=20 353-1-6770438 {BR} email: {A=20 href=3D"mailto:creid-at-tcd.ie"} creid-at-tcd.ie {/A} {/FONT} {/DIV} {/BODY} {/HTML}
Have a look in our links page http://www.proscitech.com.au/links.htm and find the category "Mail lists" using the find function. This section includes a couple of the listservers you are interested in. If others exist I would like to learn about these too. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Thursday, 6 August 1998 14:34, charles j day [SMTP:wa5ekh-at-juno.com] wrote:
} -------------. } } Are there other SEM, TEM, AFM,STM, EDS,WDS, Materials } Analysis } and Characterization Listserers? } JD } } __________________________________________________________ } ___________ } You don't need to buy Internet access to use free Internet } e-mail. } Get completely free e-mail from Juno at } http://www.juno.com } Or call Juno at (800) 654-JUNO [654-5866]
} I know this has been covered in the distant past, but I never wrote it } down. We have an open house coming up and I would like to have a } micrograph from a CD for show and tell. Could someone tell me the } sample prep steps to do this? I've never done it before.
We did something like this years ago. I can't put my hand directly on the records for the moment, but I think it went like this. The problem was that the dimples are not at the top of the CD, but buried inside with a metallic layer to reflect. As memory serves me, what we did was to cut out a small area of the CD, and then bed it shiny side down on some rapid cure Araldite. After this hardened, one could then flick off the piece of CD, leaving the shiny layer on the Araldite, and the dimples appeared as troughs in the surface that was exposed. This was then sputter coated with gold in the usual way, and examined under SEM with the direction of the grooves or dimple at the "magic angle" (put in flat, rotate so the the grooves run at 45^ to x and y, and then tilt z by 54^. I think one could also look at the remaining metallized area on the Aralidite, again gold coating.
There may be inaccuracies after such a long time of recall, but I think this will give you a general principle to work on.
Good hunting!
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Could you help me with ideas finding someone who would design a small lens system for a microscope/CCD relay lens? I'm in san Jose, California. Thanks! Julian Ortuondo
Hi Scott, I took a damaged writable cd and cut a wedge out, plastic and all. The foil peels off easily. I then mounted it on a stub with double sided carbon tab. The long line is called an "atip". This is the tract the lases follows. Then there are "lands" and "pits". These are the spaces(lands) between the laser burns(pits). Hope this helps.
Walck. Scott D. wrote: } . } } I know this has been covered in the distant past, but I never wrote it } down. We have an open house coming up and I would like to have a } micrograph from a CD for show and tell. Could someone tell me the } sample prep steps to do this? I've never done it before. } Regards, Gregory Rudomen Technical Specialist University Microscopy Imaging Center S.U.N.Y. Stony Brook 516-444-3126 Greg-at-umic.sunysb.edu
M. E. Haine, et. al. (Brit J. Appl. Phys. Vo,l 3, p. 40, '52; Vol 9, p. 482, '58; J. Brit. I.R.E. Vol. 17, p. 211, ' 57) made extensive studies of the effects of temperature, vacuum, and gun geometry on the life and brightness of tungsten filaments. This work is summarized in Ch. V! of Haine's book "The Electron Microscope", Interscience, 1961.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Try 'painting' the suface of your specimen with either 1% methylene blue or 1% methyl green. Both are aqueous solutions.
-- Begin original message --
I'm looking for a method to stain tissue en bloc so that I can see it while I'm cryo-sectioning -- something with just a hint of color.
The tissue is frog saccule, which is small and so translucent that I can't see it as I approach it through the matrix, which is OCT. The tissue is destined for immunocytochemistry, so antigenicity needs to remain preserved. (It would be helpful to find a stain that may be used for other tissue types as well -- mouse retina and chick utricle, for example.)
I was told that methylene blue may be used, but I've been unable to find anything in my reference texts.
I would be grateful for any suggestions.
Thanks,
Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu
Center for the Study of the Biology of Hearing and Deafness Central Institute for the Deaf 818 S. Euclid Ave. St. Louis, MO 63110
-- End original message -- regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
I've lived with plastic lenses for decades. Any attempt at polishing wouldn't be worth the effort. The time you take, and the *extreme* care needed in polishing isn't worth effort. Remember, you not only have to eliminate the scratches, you have to retain your prescription. This can be difficult, especially if you have any prism or astigmatism corrections. Also the front and rear surface curvatures are not likely to be the same (you have concavo-convex lenses like most?), and the optical center is likely to be nearly as thin as the lenses can be safely made already. Particularly if they're safety lenses.
Only $100? (LeHigh doesn't pay for glasses?) If your lenses aren't all that thick, get glass lenses and lessen the scratching problem. Glass is cheaper, and you'll have to put off the getting the Miata for a shorter time. 8-)
Phil
} . } I have a pair of glasses (polycarbonate lenses) which have become } scratched. The opticians I have consulted say that there is nothing to be } done except buy new lenses. The scratches are not deep. They are very } fine but, since they are in the middle of the lenses, they render the } glasses nearly useless. } } Surely with all the expertise this community has in polishing materials of } every kind, someone can tell me how to save $100. } } Alwyn Eades } . } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvannia 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net or poshel-at-hotmail.com
I make a relay lens for microscopes and ccds. What are your requirements?
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
Dear Scott, I did this years ago, and as I recall, I cut a one centimeter square from the CD, put it in tri-chorethane overnight to dissolve the plastic, then fished out the very thin Al foil that was released and put it on a shiny graphite stub. It wrinkles horribly and the little grooves are hard to see until you reach 10,000X mag, but it is possible. This is for ressed CD's, the CD-R's you make yourself are completely different. You wrote: } } I know this has been covered in the distant past, but I never wrote it } down. We have an open house coming up and I would like to have a } micrograph from a CD for show and tell. Could someone tell me the } sample prep steps to do this? I've never done it before. } } -Scott Walck } } Scott D. Walck, Ph.D. } PPG Industries, Inc.
Good luck, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Alwyn, A fine metal polish: Brasso, Wenol, etc. should do the trick You wrote: } . } I have a pair of glasses (polycarbonate lenses) which have become } scratched. The opticians I have consulted say that there is nothing to be } done except buy new lenses. The scratches are not deep. They are very } fine but, since they are in the middle of the lenses, they render the } glasses nearly useless. } } Surely with all the expertise this community has in polishing materials of } every kind, someone can tell me how to save $100. } } Alwyn Eades } . } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Hello Again, Does anyone have experience with B-gal staining?=20 (5-Bromo-4-Chloro-3-indolyl-B-galactosidase), made in solution with 5mM potassium ferricyanide, 5mM potassium ferrocyanid= e and 2mM MgCl2 in PBS.
Specifically is it electron dense, or just something used at the LM? Thanks again, Linda M. Fox lfox1-at-wpo.it.luc.edu
Does anyone have a working recipe for Araldite 502?=20
Today I rec'd tissues, processed by another lab that followed a protocol = that stated "infiltrate in Araldite". They have the tissues in a single solution of Sigma A-3058, Araldite 502.
Any suggestions as how long to reinfiltrate in a complete recipe before = embedding? ( sm. int. aprox.1mm thick)
Also, we have 10 yr+ Araldite unopened in the lab. Does this go bad after = an extended shelf life??
Thanks as always... Linda Fox lfox1-at-wpo.it.luc.edu
A slightly different approach that I use is to mix a little stain (Toluidine Blue or whatever is handy) with the OCT. It's like negative staining if the tissue is light in color. Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
Hi-pressure bottled nitrogen for SEM venting is indeed the best straightforward solution. However, special precaution should be taken to avoid a pressure rise in the chamber, which may cause the implosion of the window of the EDS detector. I would like to draw your attention to a point that, if overlooked, may result in severe damage.
EDS detectors, with either Be or ultra thin polymer windows, will withstand a pressure difference of up to 1.2 bars, as declared by most maufacturers. In order to avoid the possibility of an accidental pressure rise in the chamber beyond this value we installed a sensitive pressure regulator, operable in the 0.05 - 2 bar range, on the nitrogen feed line. The salesman who sold us the regulator advised to set it to 1.2 bar, asuming that the pressure indicated is the absolute pressure. Unfortunately, this is not the case. The indicated pressure is the gauge (manometric) pressure, namely the pressure above the ambient pressure. Had we set it to 1.2 bar, the regulator would have allowed pressure build-up of 2.2 bar, which may ruin the EDS window. The correct value to set is 0.2 bar.
I do believe this seemingly trivial communication may be useful to some of you folks.
I am trying to do TEM on some marine sponges. They are looking at a symbiotic relationship with some algae . It was fixed in 2% glut dehydrated in ETOH and embedded in med - hard Embed 812. The problem is the tissue is tearing up the sections, due I suspect to the calcium spicules, to the point that I can't even get thick sections. I asked if we can decalcify the tissue first but they felt it might damage the tissue and the correlation of the two organisms.
My library access is medical so I turned up nothing on searches. 1. Has anyone had experience with sponges and TEM? 2. how about the effect on a diamond knife (sharpened for biological work)? Thanks for any help.
I forgot to mention that any polishing will also destroy any lens coatings, and all or almost all plastic lenses have such coatings-UV, anti-reflection, and anti-scratch.
Phil
} Alwyn, } } I've lived with plastic lenses for decades. Any attempt at polishing } wouldn't be worth the effort. The time you take, and the *extreme* care } needed in polishing isn't worth effort. Remember, you not only have to } eliminate the scratches, you have to retain your prescription. This can be } difficult, especially if you have any prism or astigmatism corrections. } Also the front and rear surface curvatures are not likely to be the same } (you have concavo-convex lenses like most?), and the optical center is } likely to be nearly as thin as the lenses can be safely made already. } Particularly if they're safety lenses. } } Only $100? (LeHigh doesn't pay for glasses?) If your lenses aren't all that } thick, get glass lenses and lessen the scratching problem. Glass is } cheaper, and you'll have to put off the getting the Miata for a shorter } time. 8-) } } Phil } } } . } } I have a pair of glasses (polycarbonate lenses) which have become } } scratched. The opticians I have consulted say that there is nothing to be } } done except buy new lenses. The scratches are not deep. They are very } } fine but, since they are in the middle of the lenses, they render the } } glasses nearly useless. } } } } Surely with all the expertise this community has in polishing materials of } } every kind, someone can tell me how to save $100. } } } } Alwyn Eades } } . } } Alwyn Eades } } Department of Materials Science and Engineering } } Lehigh University } } 5 East Packer Avenue } } Bethlehem } } Pennsylvannia 18015-3195 } } Phone 610 758 4231 } } Fax 610 758 4244 } } jae5-at-lehigh.edu } } }}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ } Philip Oshel } PO Box 620068 } Middleton, WI 53562 } (608) 833-2885 } oshel-at-terracom.net } or poshel-at-hotmail.com
Microscopy-at-MSA.Microscopy.Com I teach high school biology, anatomy and physiology and introductory bilingual life sciences. I have tried every source I can find to locate posters of microscopic iimages. Students are fascinated by them and as is well known, "a picture paints a thousand words". I have yet to be able to find a source of useful scientific images for my classroom. You only need so many nature posters. If anyone can direct me to a source I will be grateful. Robin Schaeffer Kodiak, Alaska 99615 honey1-at-ptialaska.net
Robin, I happen to have a series of 16 prints of high resolution electron microscope images by David Scharf. You may view them on my web site: www.microscopy- today.com Don Grimes, Microscopy Today
} On Fri, 7 Aug 1998, Linda Fox wrote: } } } Does anyone have a working recipe for Araldite 502? } } Yes but I can't get back to the lab for a few days. You } will need NMA, DDSA and DMP-30 for my recipe. } } } Also, we have 10 yr+ Araldite unopened in the lab. Does } } this go bad after an extended shelf life?? } } Mine didn't. } } Kal
But watch out for bad catalyst, particularly if you use DMP 30! You can assume that it's gone bad.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
One of my pet peeves...You can go out to any major bookstore and find a wealth of interesting material (books, magazines) about telescopes, astronomy, photography, however, try to find something about microscopy. At a number of stores you can find poor to pretty good telescopes. Celestron markets pretty decent telescopes in the $500 to $1000 range. Have you ever seen anything but a cheap plastic toy microscope sold in retail shops?
It seems to me that there could easily be a major consumer market for by periodicals and instruments that are good quality. Considering the relative simplicity of the microscope it should be possible to design a $100-500 microscope with metal base, glass optics, a substage condenser and iris diaphragm. A video camera attachment an i/o card to allow for computer capture and image manipulation would also be a relatively inexpensive affair.
Hi: A couple of years I was called to examine some CDs lookin for defects. I used 25% NaOH/DI water solution to disolve the aluminum layer, after one night the top plastic layer floated off and I careful decanted off the freed plastic layer. I then was able to gold coat the cleaned specimen and examine it succesufuly with our SEM. You can also remove most of the top coating with a solvent (I think I used 50/50 acetone toluene) which leaves the aluminum layer which can then be looked at (gold coating improves the resolution. Terry Ellis
Hi: A couple of years I was called to examine some CDs lookin for defects. I used 25% NaOH/DI water solution to disolve the aluminum layer, after one night the top plastic layer floated off and I careful decanted off the freed plastic layer. I then was able to gold coat the cleaned specimen and examine it succesufuly with our SEM. You can also remove most of the top coating with a solvent (I think I used 50/50 acetone toluene) which leaves the aluminum layer which can then be looked at (gold coating improves the resolution. I forgot! You do have to score a line around the edge of the CD to give the NaOH room to attack the metal layer. Terry Ellis
actually you can get an onld microscope like a B&L for theat price range and put the ccd camer on it. edmund makes a nice adaptor that will do the job.. Remember the old vcr recorders that used to sit on a strap and were sort of portable? They had seperate cameras that range from an easy conversion to a conversion via. hacksaw and some adaptor making. A pity you are not in Arizona, I have some old scope bodies that would work nicely for what you want to do..
I am going to also have some duplicate microscope books for trade and failing that I will sell some. Anyone that has ian interest drop me email. thanks Ed Sharpe Archivist SMECC
Hear hear! I couldn't agree with Jim more. I would love for my niece and nephew (and grandchildren when I have any) to be able to have a decent microscope of their own. They are at that age now where the mysteries of the world are becoming so inviting! It's a pity that there aren't any $100-$500 scopes available to kindle the interest of young minds in the science of microscopy.
} One of my pet peeves...You can go out to any major bookstore and find a wealth } of interesting material (books, magazines) about telescopes, astronomy, } photography, however, try to find something about microscopy. At a number of } stores you can find poor to pretty good telescopes. Celestron markets pretty } decent telescopes in the } $500 to $1000 range. Have you ever seen anything but a cheap plastic toy } microscope sold in retail shops? } } It seems to me that there could easily be a major consumer market for by } periodicals and instruments that are good quality. Considering the relative } simplicity of the microscope it should be possible to design a $100-500 } microscope with metal base, glass optics, a substage condenser and iris } diaphragm. A video camera attachment an i/o card to allow for computer } capture and image manipulation would also be a relatively inexpensive affair. } } Any entrepreneurs out there? } } Jim Harper
Jim -
There ARE good scopes and books out there! And the science store in the small town near me carries some of them; you must be unlucky. I can refer you to lots of mail-order sources; I have a page of phone numbers that I can Email to you (that page will appear on an expanded Project MICRO web page soon). Almost all of the scopes in that price range are Chinese, from a limited number of importers, but they're sold under lots of brand names.
Don't assume that a compound scope with condenser is the "best" for a beginner; a monocular dissecting scope is better for the youngest folks. You'll find detailed comments in MSA's new middle school manual, "Microscopic Explorations". Ordering info for it (and ~100 books, videos, & CD-ROMs) is in the MICRO bibliography - address below.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Ah, you are proving my point. There are obscure nooks and cranies that contain a wealth of information where microscopists can find them--but not in the main stream where the average consumer can find them.
I agree a stereo microscope is wonderful--probably my next major scope purchase--but can I find one at Nature's Wonders in the mall? (I can find at least 5 different telescopes there.)
I guess my point is that Nikon, Zeiss, etc. seem to be missing this market--or if there were decent microscopes offered in this price range would they no longer be able to sell a $10,000 stereoscope into the average business?
MICROFAB-at-aol.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Ah, you are proving my point. There are obscure nooks and cranies that } contain a wealth of information where microscopists can find them--but not in } the main stream where the average consumer can find them. } } I agree a stereo microscope is wonderful--probably my next major scope } purchase--but can I find one at Nature's Wonders in the mall? (I can find at } least 5 different telescopes there.) } } I guess my point is that Nikon, Zeiss, etc. seem to be missing this market--or } if there were decent microscopes offered in this price range would they no } longer be able to sell a $10,000 stereoscope into the average business? } } Jim
Jim,
We are trying to produce consumer level microscopes, but interest (and sales are slow) and this makes no economical sense for us. Check out the new Naturescope; it is a stereo for under $400......
} We are in need of a good atlas identifieng the ultrastructure of viruses. } does anyone have any suggestions. } } } Christine Lee , } Veterinary Pathology, } University of Queensland } Christine Lee, } Veterinary Pathobiology, } University of Queensland. } C.lee-at-mailbox.uq.edu.au } } } ---------------------------------------------------------------------- Dr Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: GPO Box 475 ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 (0)2 6249 2743 |Australian National Univ. FAX 61 (0)2 6249 4891 |Canberra, Australia 2601 http://online.anu.edu.au/EMU/home.htm
As many subscribers probably are aware, the Microscopy Society of Southern Africa (MSSA) is bidding to host the 15th International Congress on Electron Microscopy (ICEM) in Durban, South Africa, from 1 - 6 September 2002.
Presentations by societies bidding to host the 15th ICEM, and voting on the venue, takes place during the 14th ICEM in Cancun, Mexico, later this month.
Anyone wishing to have information on the Southern African proposal is invited to visit our web site (http://www.ru.ac.za/affiliates/emu/icem2002.htm) and associated links, or may contact me directly.
Those attending the 14th ICEM in Cancun are welcome to visit the Microscopy Society of Southern Africa's booth where information will be available about the bid for the 15th ICEM as well as other information about the Society and microscopy in general in Southern Africa.
Looking forward to meeting as many as possible of you in Cancun.
Best regards,
Robin Cross Vice-President : MSSA
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
I am looking for an antibody against mouse! alpha-fetoprotein. We would like to perform immunohistochemistry on mouse tissue. Has someone made positive experiences with any antibody against this protein?
I have received over the e-mail what purports to be a TIFF file of an AFM image. However, it has been "7 bitted" for transmission using something called Bin Hex 4.0, (on a Mac, I think), and needs to be decoded using the same. Does anybody know about this beastie, and if it can work on a PC?
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I agree that the relative cost of a microscope for kids is high. For example, I can buy a pair of binoculars for $25, which contains more than twice the optics (all achromatic glass elements) and roughly the same mechanical components as a beginners compound microscope. The difference is that there is a big market for binoculars, and not for microscopes. One idea might be to develop a kit for converting a pair of cheap binoculars into a microscope (see patent 3,804,486 by Gerrit A. Van Extl and Alfred A. Akin, Jr.; assigned to Bausch & Lomb).
However, you can still buy a good compound microscope (achromatic glass elements, DIN objectives and eyepiece) for under $150 from a well known mail order scientific equipment company. I have used this scope with kids and it is great.
Several years ago I had a month-long microscope show at my local science center with the goal of getting people to buy low-cost microscopes for their kids. I felt that the cost of the microscope was a definite hurdle, but more significant was the lack of good books describing microscope activities for kids. Most (but not all) of the books readily available are plain stupid and not helpful. Interestingly, my local library has an excellent technical section with a number of turn-of-the-century books on microscopy (Marvels of pond-life; or, A year's microscopic recreations among the polyps, infusoria, rotifers, water-bears and polyzoa by Henry J. Slack, 1897 and Hunting under the microscope by Sir Arthur E. Shipley, 1928). Microscopes would be more popular if books like these were available today.
Everett Ramer Federal Energy Technology Center Pittsburgh, PA
While my EM Unit caters strictly to research, I am often asked whether I would agree that the usefulness of the EM in the clinical setting is in a major decline. It is pointed out that diagnostic EM, especially in the area of tumors, is rapidly being replaced by immunocytochemistry at the LM level. In addition, tumor cells do not as a class exhibit ultrastructural changes that can tell us much in the way of significant information anyway. I was wondering if you might have an opinion on this observation or a reference(s) where I could check this out.
Hi..having a problem with powder examination; have never before looked at powders (TiO2 in this case). Used conducting carbon adhesive pads to fix sample to stub but not much stayed on and although it coated ok most of sample came off whilst pumping out in the sample exchange chamber of our Hitachi S-800...How can i get round this..is there some sort fo fixative spray? Cheers Alan. Dr Alan Templeton Centre for Physical Electronics and Materials SEEIE South Bank University 103 Borough Road, London SE1 0AA TEL 44 171 815 7521 /7571 FAX 44 171 815 7599 email templea-at-sbu.ac.uk
My Leica paraffin tissue embedding station (essentially a hot wax dispenser) has a broken heating plate and Leica wants $3600 to repair it. I can get a new Leica for $6400 but would like to consider alternatives. Does anyone know of a different brand? I don't need a tissue processer (to run the specimen thru fix/rinse/ethanol/xylene and infilitrate in wax). I just need something for the final placement of tissue into molds. TIA. Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Linda We have been using this resin in the research lab and up in the path EM lab for over 15 years. Their formulation was based on Luft's work I believe. It is one of the most viscus resins so we take a long time infiltrating. Our last dehydration / dealcoholization step is propylene oxide "PO" (yeah I know it's toxic) 3x 10 min. we then replace with a 50/50 mix of complete "comp" resin and PO The initial resin mix is; (27 ml 502, 23 ml DDSA. if you warm the bottles to 40-50 degrees C it stirs easier). From this mix an aliquot is poured out and a 0.2%volume of DMP-30 is mixed in. We call this "comp". (Some people substitute BDMA for the DMP30 but I didn't see an infiltration or cutting advantage and you sue twice the amount as DMP30.) This 50/50 mix is left capped over night. The next day we replace the 50/50 with 100% fresh comp for two hours, then embed into gelatin capsules or flat molds. The 100% infiltration and embedding resin should be made up that day not the previous days resin (it can be stored in the fig. and used for 50/50 the next time, if that is not too long, say a couple weeks.)
In regards to OLD DMP-30.......they use it up pretty fast but buy it volume so the last bottle is probably a year old. It is an anhydrous chemical and the suppliers suggest a 3 month shelf life but obviously it can be extended. I guess like some of the other topics we discuss ie stain and fixative life its how much work vs precision, and whether you can redo a grid or experiment. We're good but usually not picky, if in doubt order fresh stuff its cheap. Lately I have switched over to the epon 812 group of resins, as they are much less viscus and cut just as well. Good luck with your work.
The Call for Papers for the International Conference on Metallurgical Coatings and Thin Films -1999 in San Diego is out. Details of the meeting can be found at the following web site.
http://home.vacuum.org/icmctf/icmctf.html
Of specific interest to physical science researchers on the Microscopy Listserver is Symposium F. There is an emphasis this year on scanning probe microscopy techniques. Session F4/B3 concentrates on Microstructural, Microanalytical and Imaging Characterization of thin films. I've included the Symposium F and the F4/B3 session synopses below. Submission of abstract can be done over the Internet.
Symposium F Coating and Thin Film Characterization Symposium Chairs: John T. Grant, Research Institute, University of Dayton, Dayton, OH 45469-0168 Phone: (937)255-6603; Fax: (937)258-8075; e-mail: grantjt-at-ml.wpafb.af.mil Hans J. Steffen, Mannheim University of Applied Sciences, Mannheim, Germany Phone: (49)621-292-6543; Fax: (49)621-292-6420; e-mail: steffen-at-fh-mannheim.de
http://home.vacuum.org/icmctf/symposium f.html
OBJECTIVES: This symposium focuses on applications and recent advances in coating and thin film characterization. Moreover, it deals with progresses in the fundamental understanding of film growth processes and the elementary structure - properties relations by analytical methods, especially but not exclusively in the fields of carbide, oxide, nitride, and DLC coatings. It is the intention of the F symposium to create an analysis group which works on hard coatings and deposition techniques for the discussion and exchange of ideas and procedures for the elucidation of growth processes and mechanisms. These topics are of particular importance as emerging deposition processes and innovative processing techniques are employed to produce thin films and coatings with unique mechanical, chemical, physical, and microstructural characteristics. Of special interest are analytical and characterization techniques and methods, including numerical evaluation procedures like factor analysis etc. to adequately describe these coatings and thin films during and after the deposition. This symposium also addresses the unique analytical challenges in the investigation of functionally gradient, multilayer, nanocrystalline, heterogeneous and composite coatings. Nondestructive and in situ characterization of all kind of coatings are also of special interest. Hence, a particular focus will be on X-ray diffraction analysis.
In 1999, Symposium F will highlight applications of all scanning probe microscopy techniques (AFM, STM, etc.) and imaging methods. These techniques have wide applicability for characterizing state of the art coating materials and thin-film architectures. The session includes topics covering theory, experiment, and sample preparation. Submissions are welcome on characterization of: mechanical, chemical, and structural properties of thin films and coatings such as friction, wear, adherence, topography, roughness, hardness, phases, and chemical forces.
Invited Speakers: Michael Serry, Digital Instruments,Recent Technologies and Advances in Thin Film Characterization Using Atomic Force Microscopy and K. Satori, Sony Corporation, Japan,Surface Roughness Development During Sputter Depth Profiling of Semiconductor and Metal Thin Films determined by AFM
F4/B3. Microstructural, Microanalytical and Imaging Characterization. Session Chairs: Siegfried Hofmann, National Research Institute for Metals, Japan and Scott D. Walck, PPG Industries. This joint session solicits papers covering advanced characterization of the micro- and nano-structure of coatings and thin films. This session will focus on recent developments in new and established techniques for microstructural characterization and imaging, with emphasis on surfaces and interfaces. Methods include but are not limited to high resolution and analytical TEM, STM/AFM, EPMA, XRD, SEM, grazing X-ray reflectivity (GXR) and other spatially resolved imaging and elemental mapping techniques. Invited Speakers: Isao Kojima, National Institute of Materials and Chemical Research,High Resolution Thickness and Interface Roughness Characterization in Multilayer Thin Films by Grazing Incidence X-ray Reflectivity, Larry F. Allard, Oak Ridge National Laboratory,Materials Characterization Via the Internet, and Kannan Krishnan, Lawrence Berkeley National Laboratory,Magnetism and Microstructures in Thin Films, Multilayers and Nanostructures.
Hello.......we are starting to do Fluorescent In Situ Hyb's of different bacterial species. My basic question is, can I re-use the slides? I use Cell-Line teflon coated slides with 6 wells, that have been cleaned with 10% KOH and then subbed with 0.1% gelatin and 0.01% chromium potassium sulfate.
Thanks, Chris Odt ---------------------------------------------- Chris Odt U.S. Dairy Forage Research Center, USDA-ARS 1925 Linden Drive West, Madison WI 53706 FAX 608-264-5147, Phone 608-264-5320
My Life According to Molecular Biology : DNA} RNA} Protein} Traits} One Spirited Red-headed Daughter
email: clodt-at-facstaff.wisc.edu "Live as though your hair is on fire....even if its thinning" Ann Onymous ----------------------------------------------
You can get some very nice posters/calendars from a number of vendors that have beautiful micrographs in them. Consider contacting Gatan, Buehler, Nikon, Olympus, Leica, Polaroid, and the different EM manufacturers for some. (This is not a complete list, but some that I know have had these in the past.) Ask them if they might have a collection of old posters and calendars as well as the current crop. I think that you will be very impressed with what some of the vendors have.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Microscopy-at-MSA.Microscopy.Com I teach high school biology, anatomy and physiology and introductory bilingual life sciences. I have tried every source I can find to locate posters of microscopic iimages. Students are fascinated by them and as is well known, "a picture paints a thousand words". I have yet to be able to find a source of useful scientific images for my classroom. You only need so many nature posters. If anyone can direct me to a source I will be grateful. Robin Schaeffer Kodiak, Alaska 99615 honey1-at-ptialaska.net
Alan, You didn't say what you are examining the powders for, but here are a = couple methods we use for powders: 1. We use a clear double-stick tape (about =BC" wide). Cut a small = piece of tape, and stick to s sample stub. Peel off the top masking layer of = the tape, and press the stub (tape side first) into a sample of the powder. Lightly blow off the stub w/ filtered dry N2 to remove loosely adhered particles. Gold coat (15-20 nm) and examine. We haven't had good luck = w/ the adhesion of the carbon pads.
2. If you are doing feature analysis on the particles and don't want agglomerations of particles, we mix 0.5 gm of sodium pyrophosphate in 1 liter of DI water. This mixture has a shelf life of 24 hours. Put approximately 0.02 gm of powder (a small bit on the end of a tooth = pick) into 40mL of this solution. Mix solution very well and withdraw a = small amount using a syringe. Make sure you mix well to keep larger = particles in suspension. Deposit a drop of the solution onto a highly polished = stainless steel coupon. Evaporate the water in an oven at 100F and examine. The sodium pyrophosphate serves as a dispersion agent to reduce particle agglomeration. You probably won't have to gold coat the sample with = this method.
John Giles Principal Materials Engineer Honeywell Space Systems Clearwater, FL
} Hi..having a problem with powder examination; have never before looked = at powders } (TiO2 in this case). Used conducting carbon adhesive pads to = fix sample to stub but not } much stayed on and although it coated ok most = of sample came off whilst pumping out in } the sample exchange chamber of = our Hitachi S-800...How can i get round this..is there } some sort fo = fixative spray? } Cheers } Alan. } Dr Alan Templeton } Centre for Physical Electronics and Materials } SEEIE
Beta-galactosidase cytochemistry (with X-gal) is usually done at the LM level. However, here are some references in which the activity is viewed by electron microscopy:
Engelhardt JF, Allen ED, Wilson JM, 1991. Reconstitution of tracheal grafts with a genetically modified epithelium. Proc Natl Acad Sci USA 88:11192-11196.
Franklin RJM, Barnett SC, 1991. The electron microscopic appearance of the beta-galactosidase reaction product. Acta Neuropathol 81:686-687.
Liu HS, Cardell, EL, Stambrook PJ, Cardell, RR, 1991. Bacterial beta-galactosidase expression in cultured mammalian cells: light and electron microscope analysis of epon sections. Anat Rec 229(4):54A (abstract).
Loewy AD, Bridgman PC, Mettenleiter TC, 1991. Beta-galactosidase expressing recombinant pseudorabies virus for light and electron microscopic study of transneuronally labeled CNS neurons. Brain Res 555:346-352.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
} Does anyone have experience with B-gal staining? (5-Bromo-4-Chloro-3- } indolyl-B-galactosidase), made in solution with 5mM potassium } ferricyanide, 5mM potassium ferrocyanide and 2mM MgCl2 in PBS. } } Specifically is it electron dense, or just something used at the LM? } Thanks again, } } Linda M. Fox } lfox1-at-wpo.it.luc.edu
Scott- previously I produced SEM samples of CD's to evaluate the etching or burning from the laser involved in the writing process. simple steps (as long as you understand the CD will be destroyed) immerse the CD in liquid nitrogen (other cryo-liquids will probably work also) for approx. 30-60 sec. You will hear it start to crack. Remove the CD with tongs, slap it down onto a hard surface (lab bench, desk) or strike it while lying on the hard surface with a hammer. The plastic/metal interface will sheer, leaving a conductive metal to examine under the SEM. I usually used carbon sticky tabs to mount, no sputtering was necessary. Good Luck -Mike
You first need to determine the composition of the spicules. Are they only calcium? Many groups of sponges have both calcareous and silaceous spicules. If they are indeed only calcareous, I'd try one of the block-face decalification procedures. These can be found in the histology and especially bone and tooth histology literature, or email me and I can give you the address of a person or two who does this.
The diamond knife might work, but you likely will need to get the knife resharpened. Ask them if they need TEM sections...perhaps SEM would answer the questions they're asking as well or better.
Phil
} } I am trying to do TEM on some marine sponges. They are looking at a } symbiotic relationship with some algae . It was fixed in 2% glut } dehydrated in ETOH and embedded in med - hard Embed 812. The } problem is the tissue is tearing up the sections, due I suspect to the } calcium spicules, to the point that I can't even get thick sections. I asked } if we can decalcify the tissue first but they felt it might damage the } tissue and the correlation of the two organisms. } } My library access is medical so I turned up nothing on searches. } 1. Has anyone had experience with sponges and TEM? 2. how about } the effect on a diamond knife (sharpened for biological work)? } Thanks for any help.
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net or poshel-at-hotmail.com
Linda- we currently use this recipe: Araldite 502 15ml Eponate 12 (or equivalent) 25ml DDSA 55ml DMP-30 (catalyst)(1.5%) 1.25ml
there are several mixtures (of varying hardness) in print, check any Hayat or Glauert book on basic principals in EM. as for reagents that are 10 years old (this stuff is cheap, your time is not) buy new resins, plastics, DMP-30, etc. -Mike
On Fri, 7 Aug 1998, Linda Fox wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Microscopy Friends, } } Does anyone have a working recipe for Araldite 502? } } Today I rec'd tissues, processed by another lab that followed a protocol that stated "infiltrate in Araldite". } They have the tissues in a single solution of Sigma A-3058, Araldite 502. } } Any suggestions as how long to reinfiltrate in a complete recipe before embedding? ( sm. int. aprox.1mm thick) } } Also, we have 10 yr+ Araldite unopened in the lab. Does this go bad after an extended shelf life?? } } Thanks as always... } Linda Fox } lfox1-at-wpo.it.luc.edu }
A faculty member here has asked me to look for images of 'aquatic pathogens' he may use on his teaching web site. He has mentioned such things as E. coli and V. cholerae but would like as great a range as possible. He wants pictures that he can use without problems of copyright releases etc.
He is also interested in other web sites or 'distance learning' resources that might 'provide the students with additional exposure to pathogenic microorganisms'. Personally, I try to avoid pathogenic microorganisms, but students may need the exposure.
Reply to me if you have something that could help him and I will pass along your offers.
Thanks
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu **Area code changing to 831 as of 7/11/98**
Alan Templeton wrote: ================================================ Hi..having a problem with powder examination; have never before looked at powders (TiO2 in this case). Used conducting carbon adhesive pads to fix sample to stub but not much stayed on and although it coated ok most of sample came off whilst pumping out in the sample exchange chamber of our Hitachi S-800...How can i get round this..is there some sort fo fixative spray? ================================================= Having looked at a lot of powders of this type ourselves, the first question is this: How old are the "carbon adhesive pads"? People often times don't realize it, but they do have a shelf life and with the passing of time, they do lose some of their original "tac". The temperature of their storage can have a lot to do with the rate of deterioration of the "tac", stored in a refrigerator, they will last much longer than if at room temperature.
Good fresh double sided conductive carbon sheets and tapes, such as you would find on the websites of SPI and some of our competitors, should contain sufficient "tac" to hold the particles. When applied dry, we like to apply a few "blasts" from a "duster" which serves to blow off the excess and also to embed a bit more firmly into the adhesive, the particles you want to study.
Another way is to mount the powders on our Tacky Dot™ Slides. See our website below for details. The smallest dot size presently available is 15 µm and if this is more than 4X larger than the size of the powder you have, the worst thing that will happen is that you will have some doublets and triplets on the dots. However, even if there should be more than one particle per dot, it is still a better way, often times, to characterize this kind of a powder, especially when using stage automation for large numbers of samples.
Disclaimer: SPI Supplies manufactures Tacky Dot™ Slides under license from the DuPont Company so we would have a vested interest in seeing more of them in use! And we also offer double sided adhesive conductive carbon sheets and tape as indicated above.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I am looking for a horizontal detector to fit a JEOL 100CX-II with an ASID or I need to get a Hack-Z package so I can install the high angle detector I currently have. Any help I get with respect to this matter will be greatly appreciated. Thanks, Mike Pidgeon
I have a JEOL 100CX-II with an ASID and need to generate some income with it. Preferably something that does not require accreditation. Does anyone have a suggestion on what I might do, and do you possibly have some contacts. I would appreciate any input on this subject. Thanks, Mike Pidgeon
Unlike a previous poster, I have had good luck using carbon loaded double face (actually mastic only) tape. With a finely divided insulator like TiO2, a thin well adhered layer is important so as to not break conduction paths after the conductive film has been applied. I prepare the mount by (as previously mentioned) gently thrusting the tape/mount into the powder or by "dusting" it onto the tape surface. Usually I don't blow off the excess, but instead sharply tap the mount a number of times while it is held up side down in my fume hood (many of my samples are rather "nasty").
I then will sputter with Au if well resolved morphology is the goal. If BSE and x-ray analysis is required, I (yarn) evaporate carbon instead. To meet both goals it is sometimes necessary to prepare two samples, one sputtered , the other C-evaporated.
...The Etec vacuum controller normally slams open a large gate valve causing a rush of air (N2 really) out of the chamber. Before modifying the pump-down, I lost a few 10mm cubes of fluffly ceramic insulation down the tubes:( I have installed a manual vacuum valve on my Etec so that I can gently rough
pump the chamber. This reduces the likelyhood that the coating will be corrupted.
Woody White McDermott Technology, Inc. mtiresearch.com
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Hi..having a problem with powder examination; have never before looked at powders (TiO2 in this case). Used conducting carbon adhesive pads to fix sample to stub but not much stayed on and although it coated ok most of sample came off whilst pumping out in the sample exchange chamber of our Hitachi S-800...How can i get round this..is there some sort fo fixative spray? Cheers Alan. Dr Alan Templeton Centre for Physical Electronics and Materials SEEIE South Bank University 103 Borough Road, London SE1 0AA TEL 44 171 815 7521 /7571 FAX 44 171 815 7599 email templea-at-sbu.ac.uk
by sciences.sdsu.edu (8.8.8/8.8.8/SCEC-8.8.8-AS.AR.S5) with ESMTP id OAA01654 for {microscopy-at-msa.microscopy.com} ; Mon, 10 Aug 1998 14:27:50 -0700 (PDT) Message-Id: {v03020902b1f517bb3299-at-[130.191.234.197]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
howdy all
anyone who knows where I can get a 35 mm camera for a Philips 410 please let me know offline
thanks
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Gary Take a look at the books by Dvorak and Monahan-Earley "Diagnostic Ultrastructural Pathology I, II, and III" (CRC Press 1992 ISBN 0-8493-4404-2) and then consider the usefulness of EM in diagnostic medicine.
Bob
_____________________________________________________________ C. Robert Bagnell Jr., Ph.D., Res.Assoc.Prof. Microscopy Services Laboratory Department of Pathology & Laboratory Medicine CB #7525 UNC-CH, Chapel Hill, N.C. 27599 ph 919-966-2413 fx 919-966-6718
For those interested, you can get a short term internet access account via Mr. Gaston C. Blanc at {gastonb-at-acnet.net} in Cancun. The charge is US $50 for the short term account which includes 150 hours of use. I have been told that payment is to be by cash only and that the service has to be arranged for in advance (but paid for in Cancun when you get your account set up).
There don't seem to be any local Compuserve numbers in that part of Mexico and calling the USA from there is expensive.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Greetings to all from the Inter/Micro 98 Meeting in Chicago. As some of you will remember, last year I made daily posts to this list summarizing some of the interesting papers being presented at Inter/Micro. This year I will again provide daily summaries but they will appear at the newly created MSA "Email-to-WWW Announcement Page" located at http://www.msa.microscopy.com/WWWviaEmail/EmailtoWWW.html . Please go there on a daily basis to review notes on many of the fascinating papers being presented at this, the 50th Anniversary Inter/Micro Meeting. Congratulations to Dr. Walter McCrone and all of the staff at McCrone Research Institute for one half century of fantastic meetings. May we live to see as many more!
I was wondering whether anybody has any experience with pointed W filaments in a XL-30. Ours currently has a normal W, although it is design for LaB6 operation. The problem I have with LaB6 is poor stability due to contamination build up on the Wehnelt (vac~2.5e-7 mBar) and hence short operation time between cleaning (not filament life time). The W is nice and stable and has been operating for several months without a clean, however I need a bit more brightness.
Are there any advantages to using pointed filaments over normal? What are the disadvantages? Are they worth the extra trouble?
Keith.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
I am a novice in this field, but have already made progress in this field specifically on polished sections of clinker specimens using etching and the like. We have a MEIJI microscope from which I can gather does not have excellent optics. Does anyone know anything about this brand of microscope and is it worth setting up a digital camera for it to do image analysis. Also we want to get a point counting stage. Any recommendations would be very welcome. Thank you.
Sincerely, Quirina
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
Pointed filaments have less than half the life, on average, when compared with standard filaments. They are more expensive and cause more downtime. They were one of the few means to achieve that extra little brightness 20 years ago. LaB6 give more brightness and reduce downtime a great deal. In the end, they are economical. Your Wehnelt contamination problem may be related to the temperature of your cooling water (Hong Kong in summer) or a number of other reasons. I would try to improve the vacuum. I also note that Kimball Lab6 cathodes arguably have the most robust design and can stand more overheating occasionally to remove contamination. I must declare that ProSciTech provides Kimball Physics Lab6 cathodes. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Tuesday, 11 August 1998 17:51, Keith Moulding [SMTP:mcmouldk-at-uxmail.ust.hk] wrote: } } Greetings, } } I was wondering whether anybody has any experience with } pointed W } filaments in a XL-30. Ours currently has a normal W, } although it is design } for LaB6 operation. The problem I have with LaB6 is poor } stability due to } contamination build up on the Wehnelt (vac~2.5e-7 mBar) } and hence short } operation time between cleaning (not filament life time). } The W is nice and } stable and has been operating for several months without a } clean, however I } need a bit more brightness. } } Are there any advantages to using pointed filaments over } normal? } What are the disadvantages? } Are they worth the extra trouble? } } Keith. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } Dr. K. Moulding. } } Materials Characterisation and Preparation Facility } Hong Kong University of Science and Technology, } Clear Water Bay, } Kowloon, } Hong Kong. } } FAX: (852) 2358 2451 } TEL: (852) 2358 8724 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ }
Energy Beam Sciences developed a range of pointed tungsten filaments more than 25 years ago, and we are certainly the primary source of these filaments worldwide, so my bias should be obvious. However, I am a professional manager, not a salesperson, so I will simply tell you what I know.
All pointed filaments are not created equal. True pointed filaments consist of single crystal tungsten points welded on top of a filament loop. While the emission is from the point, much higher brightness can be acheived. However, there are two caveats:
1. A clean vacuum is required. The point will burn out almost instantly under poor vacuum conditions.
2. Under the best of circumstances, pointed filament life is relatively short, because of the very small emission point. Once there is suffiecient material loss from the point, the filament behaves as an ordinary "loop."
Intermediate between standard filament loops and true pointed filaments is a class of filaments etched back at the tip to create a "scalpel" shape. These filaments will not be as bright as the true pointed filaments, but they are more stable, and filament life will be close to normal. We always advise customers new to pointed filament to try these first.
Please contact me back-channel if you would like additional information.
Best regards, steven E. slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
Having just gotten back into the SEM community following a 12-year hiatus, and while trying to get back up to speed by browsing through the archives of this Microscopy list server, I read with extreme interest the articles posted recently on the subject of the "ADEM1." In that thread, Craig Theberge gave a partial answer from his own personal experience to the question(s) that were raised, but I feel compelled to fill in more of the historical details, completing the picture from my own (albeit unique) perspective...
ADEM was the internal code name for a major new secret project which Fred Schamber headed up starting in 1985. (ADEM stood for "Advanced Digital Electron Microscope", I believe, but was merely coined as a code name and was not initially intended to be the actual name of the product.) The development team was organized as a separate company (later to be known as Tracor EBI) with it's own multi-million-dollar budget and staff (consisting of only a handful at the onset, handpicked from the Tracor-Northern R&D staff.) Temporary facilities were rented off-site in the Middleton Industrial Park (later dubbed the "Skunkworks") to house the entire top-secret operation. We ended up staying there for the next several years and even expanded the facility (renting adjacent bays) to accommodate additional staff (adding on to engineering, but also office personnel, stock, purchasing, drafting, etc.) Most people at the main Tracor-Northern plant didn't know what was going on just across town, and many didn't even know that the facility even existed. It was being kept a secret for the reasons that Craig mentioned - as a key microanalysis player at that time, we couldn't afford to lose our alliances with existing SEM manufacturers until we had perfected our own column. Fred was really going out on a limb with this, putting the company's future at risk financially, but felt he had the technical expertise to pull it off, and corporate was willing to trust him because of his many previous successes. After all, it would not have been an exaggeration to say that Fred was largely responsible for building the success that TN enjoyed by 1985, with a series of successful products dating all the way back to the NS-880.
Of course, in view of the unprecedented financial investment involved, Fred was under considerable pressure to deliver a return on investment in timely fashion and thereby prove that corporate's confidence in him had not been misplaced. The core R&D personnel involved at the inception were also quite dedicated to the success of the project, and so you can bet that for the next couple years, there were plenty of late hours and weekends devoted by all. Most of these have gone their separate ways now, but they included Mike Krummey (electronics engineering), Jorgen Rasmussen (mechanical engineering) and myself. We also had two primary consultants on the project: Dave Pearson (an experienced Etec serviceman, who brought in an old junker Etec scope which he kept running for us to use as a test bed for proof-of-concept on several hardware design issues), and Rich Lee! Yes, that's the same Rich (R.J.) Lee whom Fred later joined to produce the Personal SEM. A few other individuals were brought in from the outside as the need for certain skills became apparent - Ken Spielman, for example (who is still at Noran, by the way), was a skilled machinist hired in from the UW to hand-machine all the various sundry precision metal parts needed in the construction the first prototype. And of course there were technicians, draftsmen, purchasing and office support personnel, etc., added as time went on (and even marketing, sales and service people still later.) But it was Fred who spearheaded and masterminded the whole operation. It was his dream initially, his brilliance as a physicist which was crucial in solving several key problems along the way, his resourcefulness which kept the project on budget, and it was his drive and determination which kept the project moving ahead.
Fred had his hand in nearly every aspect of the development, but soon was spending the majority of his time focusing (pun intended) on the design of the lenses, which involved sophisticated electromagnetic field computations as well as careful selection of appropriate alloy materials for the pole pieces, etc. But despite the complexities involved in that, and with all due respect to Fred, those were among the more conventional aspects of the ADEM. What made ADEM truly unique was its huge chamber, sophisticated and fully-articulated sample stage, along with complete integration of all subsystems and digital control of each utilizing distributed microprocessor control. It was my pleasure to develop software and firmware for several parts of the latter, including the entire six-axes stage controller (consisting of microprocessors to drive each of the three translational and three rotational axes, and a "master" processor to coordinate the motions of all six axes.) In fact, microprocessors drove every subsystem of the ADEM, including not only the stage, but also the scan generator, vacuum system, EDS front end, etc., even the water valve! Dear reader, please indulge me as I proudly mention that I was responsible for the embedded software on nearly all of these subsystems, and personally wrote the code for most of them. But the whole instrument was tied together by a central processor which also performed the primary imaging functions; and the software for that part of the system was developed by a team of several other talented programmers. Of course, we were working with several bright electronics and mechanical engineers who should were responsible for some truly innovative hardware, not to mention a number other people from varying disciplines who made key contributions along the way. Anyway, suffice it to say that there were a lot of talented people involved in making Fred's dream a reality, and we all took great pride in our accomplishment. Indeed, as Earl Weltmer commented in the above-mentioned thread, it was quite a "work of art."
Timothy G. Moeller Senior Software Engineer NORAN Instruments Inc. 2551 W. Beltline Hwy. Middleton, WI 53562 tmoeller-at-noran.com
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We'd like to know what methods other labs are using to freeze down tissue for cryo-sectioning.
We're cooling down 2-methylbutane (iso-pentane) with dry ice and using the cooled liquid to freeze our tissue (which destined for immuno) in OCT. We don't have the facilities or the budget to use liquid nitrogen.
Are there methods that are better or use a less hazardous liquid? We want the tissue to freeze fairly quickly.
Thanks,
Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu
Center for the Biology of Hearing and Deafness Central Institute for the Deaf 818 S. Euclid Ave. St. Louis, MO 63110
by curly.cc.utexas.edu (8.8.5/8.8.5/cc-uts-1.20) with ESMTP id LAA05220 for {microscopy-at-sparc5.microscopy.com} ; Tue, 11 Aug 1998 11:47:50 -0500 (CDT)
Hi All, I'm tring to find a supplier of TEM grids that still offers custom photo- etching service of Au grids or screening. Has anyone used such services or could you point me in the right dirrection i.e. PolySciences, Ted Pella etc.
Alan, I'm not sure what type of analysis you are doing but if you are looking at sizing, most of the methods mentioned use a blow off or knock off. This would definitely skew your analysis. Getting a representative sampling of fine powders is a real challange. Finding a good method is very sample dependent. A good start for TiO2 would be a suspension of a powder sample in wetable water coupled with a 100% recovery on a small membrane filter. Russ
-----Original Message-----
Hi..having a problem with powder examination; have never before looked at powders (TiO2 in this case). Used conducting carbon adhesive pads to fix sample to stub but not much stayed on and although it coated ok most of sample came off whilst pumping out in the sample exchange chamber of our Hitachi S-800...How can i get round this..is there some sort fo fixative spray? Cheers Alan. Dr Alan Templeton Centre for Physical Electronics and Materials SEEIE South Bank University 103 Borough Road, London SE1 0AA TEL 44 171 815 7521 /7571 FAX 44 171 815 7599 email templea-at-sbu.ac.uk
We are constructing some microscopy chambers and I have a vague recollection of reading some clever way to eliminate the meniscus effect so that ones gets an even buffer layer in a small chamber. Anyone have an idea whether I am dreaming or not? Thanks- Dave Knecht
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
} Linda- } we currently use this recipe: } Araldite 502 15ml } Eponate 12 (or equivalent) 25ml } DDSA 55ml } DMP-30 (catalyst)(1.5%) 1.25ml } } there are several mixtures (of varying hardness) in print, check any Hayat } or Glauert book on basic principals in EM. } as for reagents that are 10 years old (this stuff is cheap, your time is } not) buy new resins, plastics, DMP-30, etc. } -Mike } Linda & Mike -
A couple of days ago I suggested DMP-30 as the epoxy component that is most likely to go bad with long storage, but I didn't explain why. DMP-30 is inactivated by water, and it WILL hydrate if it isn't stored carefully. BDMA is an equally efficient accelerator, and is much less viscous than DMP-30;it can and should be used instead of DMP-30 in ANY epoxy recipe (Glauert, A.M. Accelerators for epoxy resins. Proc. R.M.S. 22:264,1987). The persistence of DMP-30 is a historical accident.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I'm tring to find a supplier of TEM grids that still offers custom photo- etching service of Au grids or screening. Has anyone used such services or could you point me in the right dirrection i.e. PolySciences, Ted Pella etc.
Dear Listers, If anyone knows of a source for a UV lamp - Cathodeon, TW6W black UV lamp for Leica's AFS in the states, would you let me know? Thanks in advance! Rosemary
Advanced Bioconcept produces a wide range of novel, high affinity, biologically active fluo-peptides for identification & characterisation of GPCR's, which can be very valuable to reaserchers in Confocal Microscopy.
They can be used to study peptide-receptor interactions in:
We have 31 established Fluo-peptides, and many are in development. If you would be kind enough to forward your contact details (fax number preferably), we would be very pleased to send you information about these products, such as protocols, a technical data sheets, and a product & price list. We can also outline our novel philosophy on custom syntheses if you wish.
Best regards,
Jon.
Jon Pinchuk ============================ Advanced Bioconcept Ltd. 1440 St. Catherine W., #424 Montreal, Qc, Canada, H3G1R8 http://www.bioconcept.com/ ============================ tel:514-874-5434 x22 fax:514-874-9077 mailto:jon-at-bioconcept.com ============================
Try Bulb Direct: 800-772-5267. Bulb Direct carries all kinds of bulbs under the sun. They may help you.
Regards,
In C. Kim, Ph.D. Ingen Laboratories, Inc. inkim-at-ingenlabs.com www.ingenlabs.com ---------------------------------------------------------- "Explorer of Molecular Hybridization"
-----Original Message-----
John N. Wright wrote: } } } Hi All, } I'm tring to find a supplier of TEM grids that still offers custom photo- } etching service of Au grids or screening. Has anyone used such services or } could you point me in the right dirrection. } } Thanks in Advance, } John N. Wright
-- Dear Mr. Wright,
We at Ladd Research would be happy to quote you. Please give us the mesh size requirements and the quantities and we will send you a quote or call the number below to discuss it.
Thanks,
John Arnott
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
The question of venting chambers and colums with dry nitrogen, using the boil-off gas from a liquid nitrogen container, has come up once or twice previously. As I mentioned then, there is a very inexpensive way to to this. What you do is to connect the vent tube of the liquid nitrogen container to the gas inlet of the chamber with a flexible tube (ordinary polyethylene tubing seems to work quite well, and is inexpensive and easy to work with). At a convenient spot along this tubing line insert a tee joint. Obtain a large inflatable-deflatable plastic beach ball and attach it to this tee joint with a length of flexible surgical rubber tubing. Using a razor blade or sharp scalpel, cut a clean slit about 100 mm long in this surgical rubber tubing. This slit then acts as a primative, but quite effective, pressure release valve. If the cut is clean, straight, and parallel to the axis of the tubing it will normally close tightly enough so that the nitrogen that boils off the storage container will accumulate in the beach ball. If the pressure in the ball rises significantly above that of the surrounding atmosphere, however, the slit will open slightly and allow the gas to escape. When the inlet valve to the vacuum chamber is opened, the slit will remain closed tightly enough so that no atmospheric gas can enter. The gas in the beach ball will then be driven into the chamber by only the pressure of the surrounding atmosphere so that overpressurization cannot occur. A beach ball that is about 2 ft in diameter will hold about 100 liters of gas, which is enough to fill most instruments several times, and of course the gas is being constantly replenished from the liquid nitrogen container.
If you are planning to use gas from a high pressure cylinder to fill EM chanbers, then you must be careful to ensure that you truly obtain oil-free gas in an oil-free tank. Ordinary compressed gases are often pumped with oil-sealed pumps and are contaminated with oil vapors from the pump, while ordinary tanks are usually contaminated with the oil vapors carried into them in this way.
These and related matters are discussed in some detail on pp. 63 - 65 of my book, 'Vacuum Methods in Electron Microscopoy' (see. http://www.cccbi.chester.pa.us/spi/catalog/books/book48.html, or http://www.bookshop.co.uk/portland/)
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
I am attempting to cryosection starch-based foams, but they are collapsing in the TissueTek embedding medium because of the water content. Does anyone have ideas on making an embedding medium which is at least 95% anhydrous? I need the foams to keep their structure at least long enough for me to infitrate it in a vacuum(to get the medium into the air pockets) and then be frozen at about -20degC.
My "Correspondent's Report" from Inter/Micro 98 has just been posted to http://www.msa.microscopy.com/WWWviaEmail/EmailtoWWW.html . Please drop by and give it a read.
Stephen A. Shaffer MicroDataware sshaffer-at-microdataware.com
I would concur with a couple of other postings to your questions. The pointed filaments are not a good alternative. LaB6 should be the preferred cathode.
You don't give very specific details of your problem, but I see two possible problems. The first involves the manufacture of the LaB6 cathodes you use. You may be experiencing substantial amount of cathode drift after a few hours of use. This will result in a lower beam current, noisier image and a filament image that drifts to being eclipsed by apetures. In this case, I agree with another poster that Kimball Physics cathodes are quite exceptional. While you may associate this problem with the grid contamination, are you also re-centering your cathode when you clean the grid?
The other possibility is a vacuum leak. While you are able to attain and maintain a good overall vacuum, it is possible that there are vacuum leaks that direct incoming air to the cathode. Suspect the gun translation mechanism, if there is one, along with any vacuum standpipes or valves that open to the gun area. If the contamination is indeed the cause of your problems, an air leak is the most probable cause.
A properly designed and manufactured LaB6 cathode should not, on its own, produce grid contamination that would affect the imaging capability of the instrument. In fact, it would require a great deal of contamination to produce any visible effect on the imaging capability of an SEM in normally used magnification regions. If grid contamination is the actual cause of your problems, then you are either operating at the outside of the envelope for an SEM or the contamination is excessive because of vacuum systems leaks. On the other hand, you may be falsely attributing the cause to contamination where it really belongs to the poor design and manufacture of the cathodes you are using.
} Greetings, } } I was wondering whether anybody has any experience with pointed W } filaments in a XL-30. Ours currently has a normal W, although it is } design for LaB6 operation. The problem I have with LaB6 is poor } stability due to contamination build up on the Wehnelt (vac~2.5e-7 } mBar) and hence short operation time between cleaning (not filament } life time). The W is nice and stable and has been operating for } several months without a clean, however I need a bit more } brightness. } } Are there any advantages to using pointed filaments over normal? } What are the disadvantages? Are they worth the extra trouble? } } Keith. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Dr. K. Moulding. } } Materials Characterisation and Preparation Facility } Hong Kong University of Science and Technology, } Clear Water Bay, } Kowloon, } Hong Kong. } } FAX: (852) 2358 2451 } TEL: (852) 2358 8724 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Thanks for the info. I was unfamiliar with the ADEM and since I service virtually every SEM made, I was quite curious when mention was made of it. I used to work with Pearson at ETEC. I had heard that he was helping with development of a new SEM, but never knew all the connections.
I'm sorry I've never had the opportunity to work with an ADEM. From your description, it sounds like they suffered in part from the common belief of the time that microcontrollers should be used where ever possible. Another example is ARL in the late eighties with their line of x-ray fluorescence spectrometers. While large numbers of microcontrollers can reduce the actual chip count of complex circuitry, the software development, debugging and basic understanding of the circuits being replaced has proven to limit their use in many cases.
I don't intend to disparage your vocation, rather I wish to point out that unrealistic expectations are often placed on it.
The development of a new SEM is a daunting and expensive task. The major manufacturers have it much easier, generally making small evolutionary changes. Producing a true revolutionary change requires money and perseverance. Those are things that few companies understand or can weather, particularily in today's corporate climate.
While the politics of Tracor's position may have lead to ADEM's demise, it may also have been its vision. Attempting to produce a revolutionary change from an evolutionary product may have ensured that the corporate expectations would be left wanting.
} Having just gotten back into the SEM community following a 12-year } hiatus, and while trying to get back up to speed by browsing through } the archives of this Microscopy list server, I read with extreme } interest the articles posted recently on the subject of the "ADEM1." } In that thread, Craig Theberge gave a partial answer from his own } personal experience to the question(s) that were raised, but I feel } compelled to fill in more of the historical details, completing the } picture from my own (albeit unique) perspective... } } ADEM was the internal code name for a major new secret project which } Fred Schamber headed up starting in 1985. (ADEM stood for "Advanced } Digital Electron Microscope", I believe, but was merely coined as a } code name and was not initially intended to be the actual name of } the product.) The development team was organized as a separate } company (later to be known as Tracor EBI) with it's own } multi-million-dollar budget and staff (consisting of only a handful } at the onset, handpicked from the Tracor-Northern R&D staff.) } Temporary facilities were rented off-site in the Middleton } Industrial Park (later dubbed the "Skunkworks") to house the entire } top-secret operation. We ended up staying there for the next } several years and even expanded the facility (renting adjacent bays) } to accommodate additional staff (adding on to engineering, but also } office personnel, stock, purchasing, drafting, etc.) Most people at } the main Tracor-Northern plant didn't know what was going on just } across town, and many didn't even know that the facility even } existed. It was being kept a secret for the reasons that Craig } mentioned - as a key microanalysis player at that time, we couldn't } afford to lose our alliances with existing SEM manufacturers until } we had perfected our own column. Fred was really going out on a } limb with this, putting the company's future at risk financially, } but felt he had the technical expertise to pull it off, and } corporate was willing to trust him because of his many previous } successes. After all, it would not have been an exaggeration to say } that Fred was largely responsible for building the success that TN } enjoyed by 1985, with a series of successful products dating all the } way back to the NS-880. } } Of course, in view of the unprecedented financial investment } involved, Fred was under considerable pressure to deliver a return } on investment in timely fashion and thereby prove that corporate's } confidence in him had not been misplaced. The core R&D personnel } involved at the inception were also quite dedicated to the success } of the project, and so you can bet that for the next couple years, } there were plenty of late hours and weekends devoted by all. Most } of these have gone their separate ways now, but they included Mike } Krummey (electronics engineering), Jorgen Rasmussen (mechanical } engineering) and myself. We also had two primary consultants on the } project: Dave Pearson (an experienced Etec serviceman, who brought } in an old junker Etec scope which he kept running for us to use as a } test bed for proof-of-concept on several hardware design issues), } and Rich Lee! Yes, that's the same Rich (R.J.) Lee whom Fred later } joined to produce the Personal SEM. A few other individuals were } brought in from the outside as the need for certain skills became } apparent - Ken Spielman, for example (who is still at Noran, by the } way), was a skilled machinist hired in from the UW to hand-machine } all the various sundry precision metal parts needed in the } construction the first prototype. And of course there were } technicians, draftsmen, purchasing and office support personnel, } etc., added as time went on (and even marketing, sales and service } people still later.) But it was Fred who spearheaded and } masterminded the whole operation. It was his dream initially, his } brilliance as a physicist which was crucial in solving several key } problems along the way, his resourcefulness which kept the project } on budget, and it was his drive and determination which kept the } project moving ahead. } } Fred had his hand in nearly every aspect of the development, but } soon was spending the majority of his time focusing (pun intended) } on the design of the lenses, which involved sophisticated } electromagnetic field computations as well as careful selection of } appropriate alloy materials for the pole pieces, etc. But despite } the complexities involved in that, and with all due respect to Fred, } those were among the more conventional aspects of the ADEM. What } made ADEM truly unique was its huge chamber, sophisticated and } fully-articulated sample stage, along with complete integration of } all subsystems and digital control of each utilizing distributed } microprocessor control. It was my pleasure to develop software and } firmware for several parts of the latter, including the entire } six-axes stage controller (consisting of microprocessors to drive } each of the three translational and three rotational axes, and a } "master" processor to coordinate the motions of all six axes.) In } fact, microprocessors drove every subsystem of the ADEM, including } not only the stage, but also the scan generator, vacuum system, EDS } front end, etc., even the water valve! Dear reader, please indulge } me as I proudly mention that I was responsible for the embedded } software on nearly all of these subsystems, and personally wrote the } code for most of them. But the whole instrument was tied together } by a central processor which also performed the primary imaging } functions; and the software for that part of the system was } developed by a team of several other talented programmers. Of } course, we were working with several bright electronics and } mechanical engineers who should were responsible for some truly } innovative hardware, not to mention a number other people from } varying disciplines who made key contributions along the way. } Anyway, suffice it to say that there were a lot of talented people } involved in making Fred's dream a reality, and we all took great } pride in our accomplishment. Indeed, as Earl Weltmer commented in } the above-mentioned thread, it was quite a "work of art." } } Timothy G. Moeller } Senior Software Engineer } NORAN Instruments Inc. } 2551 W. Beltline Hwy. } Middleton, WI 53562 } tmoeller-at-noran.com } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Does anyone have a good method for cleaning powder metal samples that have been emmersed in oil? Specifically, these parts are saturated with transmission fluid and it is extremely difficult to examine them in the SEM without some seeping of the fluid from the interior of the part. Past attempts have included vacuum, a wide range of solvents, sonication as well as heating to burn off any remaining fluid. Any info is greatly appreciated.
Dear all, in our TEM-Lab runs a discussion about the use of uranylacetate in routi= ne secion-staining because of its radioactivity and the possible risk of ca= ncer. We are wondered about the relative 'light' safety instructions for handl= ing and disposal uranylsolutions compared to that for C-Isotopes used in special enzymat= ic reactions. 1.Is this risk negligible , if not, what intern al safety instructions exi= sts in other labs or is literature about the risk of uranylacetat in biological labs available? 2.Has anybody experience with alternative (non-radioactive) counter stains= ? Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=E4tsstrasse 25 Germany 33615 Bielefeld phone: 0521 1065592 fax: 0521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
Thank you for the kudos. And, no offense taken regarding the fact that we went overboard with the use of microprocessors in the design of the ADEM - in fact, I agree with you. Indeed, that is one of the reasons for it's short life (after all, the original poster which prompted my little expose' was trying to dispose of two "old" ADEMs already, not even 10 years old!) They are just too complex to maintain &/or modify. And, as proud as I am of my embedded code, I must admit that the fact that it is entirely proprietary and not user-programmable makes it extremely inflexible.
You are certainly correct about the development of a new SEM being "a daunting and expensive task." The ADEM development team was all too well aware of that, even from the start. And we had that truth painfully impressed upon us ever more so each day as we struggled with trying to get it done. It would be really nice to hear from Fred (Dr. Frederick H. Schamber, now with RJ Lee Instruments, as I mentioned) to fill in even more of the background. He bore the brunt of many of the criticisms which you have voiced concerning the decision to develop the ADEM, and I'm sure would be able to offer further insights into the rationale for doing so. If nothing else, I believe he would mention that there was some motivation for creating an SEM entirely made in the U.S.A., at a time when the American work ethic was being challenged by the Japanese, and blamed for the growing trade deficit. At any rate, I don't think anyone could really fault him for the decision; at least his motives were pure, even if he lacked some business savvy. And he was certainly up to the task from a technical standpoint; after all, if anybody could pull it off, Fred could - he's a uniquely talented, extremely intelligent and exceptionally energetic guy, for whom I have the utmost respect and admiration. The SEM community is blessed to still have him in their midst, even if it's just working on " evolutionary" SEMs.
Another bit of historical information (completely unrelated) that I just remembered... At the same time we were developing the ADEM, the birth of our first CONFOCAL microscope took place. Dr. D. O. Landon got a few of us involved at the "Skunkworks" off in a corner, playing on an optical table with lasers, motion controllers and other cool "toys", actually acquiring high-resolution images of biological specimens. I even wrote some TN-5500 software (in FLEXTRAN) in my spare time to control and display images from that very first confocal microscope. But of course that seemed like just a fun diversion; we didn't regard this as an important development at the time. Oddly enough, however, confocal microscopy remains a significant part of the core business here at NORAN, second only to the microanalysis instrumentation products, and confocal are the only microscopes we manufacture at all anymore. Kind of shows the value of always keeping something on the "back burner", I guess.
Timothy G. Moeller Senior Software Engineer NORAN Instruments Inc. 2551 W. Beltline Hwy. Middleton, WI 53562 (608) 836-4119 tmoeller-at-noran.com
----------
Thanks for the info. I was unfamiliar with the ADEM and since I service virtually every SEM made, I was quite curious when mention was made of it. I used to work with Pearson at ETEC. I had heard that he was helping with development of a new SEM, but never knew all the connections.
I'm sorry I've never had the opportunity to work with an ADEM. From your description, it sounds like they suffered in part from the common belief of the time that microcontrollers should be used where ever possible. Another example is ARL in the late eighties with their line of x-ray fluorescence spectrometers. While large numbers of microcontrollers can reduce the actual chip count of complex circuitry, the software development, debugging and basic understanding of the circuits being replaced has proven to limit their use in many cases.
I don't intend to disparage your vocation, rather I wish to point out that unrealistic expectations are often placed on it.
The development of a new SEM is a daunting and expensive task. The major manufacturers have it much easier, generally making small evolutionary changes. Producing a true revolutionary change requires money and perseverance. Those are things that few companies understand or can weather, particularily in today's corporate climate.
While the politics of Tracor's position may have lead to ADEM's demise, it may also have been its vision. Attempting to produce a revolutionary change from an evolutionary product may have ensured that the corporate expectations would be left wanting.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
I am going to be transiting in Miami on my way to Cancun and have been told that I would need a VISA for these few minutes in the United States. Is that really true? Kristof Kovacs
************************************************************************* Dr. Kristof KOVACS President, Hungarian Society for Microscopy Associate Professor University of Veszprem Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684 Fax: +36-(88)-328-643 *************************************************************************
We routinely use the iso-pentane and dry ice method with good results on skin biopsies. We also use ethanol and dry ice with the same results. You just have to be very careful not to let the ethanol come in to contact with the OCT. It will make it rubbery and can't be cut.
The other issue is if you need to fix the tissue you need to cryo-protect it after fixation before you embed and freeze it, or it will shread as it thaws. We use Zambonies or paraformaldehyde, then after rinsing in PBS, cryoprotect in 8.5% sucrose in PBS for two hours then embed in OCT and freeze.
Bob Derm Imaging Center U of W
On 11 Aug 1998, Jaci Lett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We'd like to know what methods other labs are using to freeze down tissue for } cryo-sectioning. } } We're cooling down 2-methylbutane (iso-pentane) with dry ice and using the } cooled liquid to freeze our tissue (which destined for immuno) in OCT. We don't } have the facilities or the budget to use liquid nitrogen. } } Are there methods that are better or use a less hazardous liquid? We want the } tissue to freeze fairly quickly. } } Thanks, } } Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu } } Center for the Biology of Hearing and Deafness } Central Institute for the Deaf } 818 S. Euclid Ave. } St. Louis, MO 63110 }
A lady replied on my query regarding MEIJI microscopes.
I would like to say thank you for taking the effort to reply and I am sorry but I have mistakenly deleted your message. Could you perhaps E-mail me again...I would like to discuss perhaps the optics in more details. I am of the opinion at the moment that the optics are not good enough to waraant equiping it with a digital camera since there are already enough problems with image analysis from a good image let alone a poor one.
And if anyone out there could make recommendations on optical microscopes. For example, I know Zeiss is supposed to be the best, with Leica and Nikon following. I want to know more than just the brandname, but I want information on the exact optical configuration and why it is better or worse than another, because suppliers seem to be too secretive, so I am asking USERS for their expert opinion based on sound experience.
I would appreciate very much any avalaible information or at least sources where I could find this kind of information.
Cherio, Quirina
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
Jaclynn, My opinion after freezing tons of tissue for four years is that 2-methylbutane cooled in dry ice is the way to go- quick,simple,cheap. I made sure the 2-mb was cooled enough by putting in a few small pieces of dry ice until the vigorous bubbling stopped or slowed almost completely (at which the temperatures are nearly equal). I also made sure that any tools were cooled beforehand. Lastly, when the tissue is plunged quickly into the cooled 2-mb, move it around gently to keep it in contact with cooled fluid. Have you considered cryoprotectants? Are you experiencing problems? "Practical Methods in Electron Microscopy, vol 13, Sectioning and Cryosectioning for Electron Microscopy," is a good reference.
-------------------------------------------------------- Winston W Wiggins, Supr. 8/12/98 10:49:56 AM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org --------------------------------------------------------
We are trying to do immunostaining of newt tissue, which, while alive, was expressing GFP. Cryosectioning is an option. I heard that prolonged fixation in paraformaldehyde will quench the GFP signal - is that true? The decision to make is whether to prefix the tissue for longer period of time and risk losing the signal or quick freeze, section and then fix for shorter period of time. Also, what would be an optimal fixative for GFP retention?
Sharing any thoughts or direct experience in this area would be highly appreciated.
Judy Trogadis Eye Research Institute and University of Toronto Toronto Hospital, Western Div. 399 Bathurst St. Toronto, Canada M5T 2S8
The program and preliminary poster for the joint NYSEM and AIF all day symposium at the Albert Einstein College of Medicine on Sept. 10 is posted at http://www.ca.aecom.yu.edu/aif/directions.htm
-------------------------------------------- Michael Cammer email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 http://www.ca.aecom.yu.edu/aif/ --------------------------------------------
We are looking for two used research quality inverted microscopes, e.g., Zeiss IM35, Nikon Diaphot 200 or later, or Olympus I50 or 70, for patch clamp recording purposes. If anyone has ones available please email: yamoahen-at-email.uc.edu, or call Ebenezer Yamoah at 513-558-4909.
I have been trying to silver enhance 1nm gold label that is embedded in LR White sections. After prolonged (90min) soaking in BBI sliver enhancing solution (SEK15), the surface particles are very large, but the enhancement is not the penetrating the plasticand getting to the gold trapped in the section. Does anyone have a method for permualizing LR White, so that enhancing solution penetration will occur?
TIA
William R.McManus Supervisor Electron Microscope Facility Department of Biology Utah State University Logan UT 84322-5305
(1.38.193.4/16.2) id AA20297; Thu, 13 Aug 1998 07:50:13 -0400 Received: by DA_EXC1.sylvania.com with Internet Mail Service (5.5.2232.9) id {Q5KXC88S} ; Thu, 13 Aug 1998 07:12:28 -0400
I'm looking for an independent failure analysis lab in the Southwestern US. The lab should be capable of performing microanalytical techniques on glasses, ceramics, metals, plastics and composites. Commercial and academic labs are acceptable. Please respond to me directly.
Thanks,
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
I have been playing around with sovelnts (diff pump stack revival) Carbon tertracloride CCl4 fumes are brilliant. It boils at 76.54 deg Centegrade. Be sure that you work in a fumehood! Is used as a degraser at Cr and Ni coating facilitys. } } Does anyone have a good method for cleaning powder metal samples that have } been emmersed in oil? Specifically, these parts are saturated with } transmission fluid and it is extremely difficult to examine them in the SEM } without some seeping of the fluid from the interior of the part. Past } attempts have included vacuum, a wide range of solvents, sonication as well } as heating to burn off any remaining fluid. Any info is greatly } appreciated. } } Wayne England } wengland-at-ortech.on.ca }
Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
Attached is the TOC for the journal MICRON for August 1998
CONTENTS LIST Date 13-JUL-1998 Page 1
Journal : Micron Volume/issue : 29/4 Year : 1998 ISSN : 0968-4328 Cover Date : August 1998
pp. iv-iv Editorial
pp. 251-259 PII S0968-4328(97)00062-0 JMIC 232 Ed. office no. E53 Microstructure of polarized electrochemical vapor deposition (PEVD) products EZ Tang, DG Ivey, TH Etsell
pp. 261-265 PII S0968-4328(97)00063-2 JMIC 233 The significance of locating and filling the canal isthmus in multiple root canal systems. A scanning electron microscopy study of the mesiobuccal root of maxillary first permanent molars DC Yu, A Tam, MH Chen
pp. 267-280 PII S0968-4328(97)00064-4 JMIC 234 Prosobranch parasperm: sterile germ cells that promote paternity? J Buckland-Nicks
pp. 281-287 PII S0968-4328(97)00057-7 JMIC 227 Ed. office no. E 52 Microstructural characterization of Au/Sn solder for packaging in optoelectronic applications DG Ivey
pp. 289-292 PII S0968-4328(97)00053-X JMIC 223 Ed. office no. E56 Events at the university of Alberta and the university of Toronto, leading to the first North-American electron microscope AF Prebus
pp. 293-296 PII S0968-4328(98)00013-4 JMIC 257 Modification of unicryl composition for rapid polymerisation at low temperature without alteration of immunocytochemical sensitivity P Gounon, J-P Rolland
pp. 297-307 PII S0968-4328(98)00011-0 JMIC 255 Optimization of phosporous localization by EFTEM of nucleic acid containing structures C Quintana, S Marco, N Bonnet, C Risco, ML Gutierrez, A Guerrero, JL Carrascosa
pp. 309-328 PII S0968-4328(98)00015-8 JMIC 259 Aspects of the structure and assembly of desmosomes IDJ Burdett
Dear Judy, Long fixation may reduce GFP signal. Paraformaldehyde fix of 30 minutes should be sufficient for most systems. However. You will lose all GFP signal if it is a free GFP in the cytosol or nucleus upon permeabilization due to extraction. You do not mention whether this is a GFP fusion protein. GFP- fusion proteins that a integral parts of membranes or structures will remain.
Joseph Goodhouse Confocal / EM Core Lab Manager Department of Molecular Biology Princeton University jgoodhose-at-molbio.princeton.edu 609-258-5432 Info and Images at http://www.molbio.princeton.edu/confocal/CF-EM-HOME.html
} -----Original Message----- } From: Judy Trogadis [SMTP:judy-at-playfair.utoronto.ca] } Sent: Wednesday, August 12, 1998 11:26 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: GFP immuno } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } We are trying to do immunostaining of newt tissue, which, while alive, } was } expressing GFP. Cryosectioning is an option. I heard that prolonged } fixation } in paraformaldehyde will quench the GFP signal - is that true? The } decision } to make is whether to prefix the tissue for longer period of time and } risk } losing the signal or quick freeze, section and then fix for shorter } period } of time. Also, what would be an optimal fixative for GFP retention? } } Sharing any thoughts or direct experience in this area would be highly } } appreciated. } } } Judy Trogadis } Eye Research Institute and } University of Toronto } Toronto Hospital, Western Div. } 399 Bathurst St. } Toronto, Canada M5T 2S8 } } phone: 416-603-5088 } Fax: 416-603-5126 } email: judy-at-playfair.utoronto.ca }
It seems most of the avid writers are conferencing, so here=20 is my contribution: This forum has dealt with UA safety several times before.=20 It is, however, very time consuming to go through a year of=20 the archives, so here are is my summary/opinion.
1 All elements above lead in the periodic table are=20 radioactive. 2 Carbon, phosphorus and other isotopes are readily=20 absorbed and incorporated in tissues. That makes these=20 "biological isotopes" more dangerous. Uranyl Acetate is water soluble and is not stored in the=20 body. 3 Whereas in mining, the insoluble Uranium particles are=20 lodged in the lung and emit radon gas for many years and=20 this makes insoluble U compounds a much greater=20 radiological hazard. 4 UAs' chemical toxicity is greater than its radiation=20 hazard - just don't ingest that stuff, it's a powerful=20 kidney poison, but happily not cumulative. 5 In general terms, the higher an element on the periodic=20 table, the more intense the electron "staining". A good=20 argument for lead, being the last non-radioactive element,=20 except that it a cumulative toxin, but we use it with care=20 (I trust). Actually I have seen lead pigments used in an=20 industrial setting (36 yrs ago), truly hair raising by=20 today's standards, but it was known then that lead was a=20 cumulative poison. 6 The (Edelmetalle) Au, Pd, Pt, Ir would seem attractive=20 alternatives for electron staining, but they are=20 non-reactive and only useful as markers or in evaporation=20 techniques. 7 Dense, high molecular number compounds have some staining=20 effects, for instance Sudan Black to show lipids, but these=20 stains are not nearly as effective as are the high atomic=20 number elements. 8 I am worried why so many people are worried. You would=20 have cause if you were dealing with the 200 litre drum=20 quantities of U "yellowcake", which has very similar=20 toxicity and radioactive attributes. 9 Consider: You are in a laboratory with fumehood and=20 gloves available, you are rather more knowledgeable about=20 these matters then armies of industrial workers and you are=20 using tiny quantities. 10 On balance I would suggest that it is rather more=20 dangerous to fly at high altitudes because of the gamma=20 radiation and it's a terrible thing to eat steak, because=20 of the high fat content and any scorched parts are=20 carcinogenic.
If you handle in a laboratory setting UA prudently, it is=20 in my opinion one of the less hazardous encounters in life.=20 I suggest that even a walk in the Teutoburger Wald, not to=20 mention urban Bielefeld is not without its dangers. Cheers Jim Darley ProSciTech Microscopy=20 PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au=20 *****
On Wednesday, 12 August 1998 23:25, B. Laube=20 [SMTP:B.Laube-at-biologie.uni-bielefeld.de] wrote: } Dear all, } in our TEM-Lab runs a discussion about the use of } uranylacetate in routine } secion-staining because of its radioactivity and the } possible risk of cancer. } We are wondered about the relative 'light' safety } instructions for handling and disposal } uranylsolutions compared to that for C-Isotopes used in } special enzymatic reactions. } 1.Is this risk negligible , if not, what intern al safety } instructions exists in other labs or is } literature about the risk of uranylacetat in biological } labs available? } 2.Has anybody experience with alternative (non- } radioactive) counter stains? } Bernward Laube } University of Bielefeld } Faculty of Biology } Department Plant Morphology and Cell Ultrastructure } Universit=E4tsstrasse 25 } Germany 33615 Bielefeld } phone: 0521 1065592 } fax: 0521 1066039 } e-mail: b.laube-at-biologie.uni-bielefeld.de } http://www.uni- } bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
hello....anybody know an easy way i.e. via a macro or similar to get a conversion from the ASTM G grain size number to the actual value in real dimensions?..is it different for the various ASTM grain size measurement types that are available ,i.e horizontal, vertical, diagonal and concentric circles. The image analysis package we have is Image Pro Plus with Materials Pro from Media Cybernetics (USA) which gives us the grain size in the ASTM G unit. I have info from the ASTM standards E1382 and E112 concerning this but there is a lot of info in there I'm not sure about. cheers. alan. ----- Dr Alan Templeton Centre for Physical Electronics and Materials SEEIE South Bank University 103 Borough Road, London SE1 0AA TEL 44 171 815 7521 /7571 FAX 44 171 815 7599 email templea-at-sbu.ac.uk
We use sodium m-periodate saturated, for about 10-20 min to etch away some of the LRwhite but I have never tried this on prelabelled samples. I would think it might strip away the gold.
bob Derm Imaging Center U of W
On Wed, 12 Aug 1998 billemac-at-cc.usu.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have been trying to silver enhance 1nm gold label that is embedded in LR } White sections. After prolonged (90min) soaking in BBI sliver enhancing } solution (SEK15), the surface particles are very large, but the enhancement } is not the penetrating the plasticand getting to the gold trapped in the } section. Does anyone have a method for permualizing LR White, so that } enhancing solution penetration will occur? } } TIA } } } William R.McManus } Supervisor } Electron Microscope Facility } Department of Biology } Utah State University } Logan UT 84322-5305 } } billEMac-at-cc.usu.edu } Ph 435-797-1920 } Fax 435-797-1575 } } }
} in our TEM-Lab runs a discussion about the use of uranylacetate in routine } secion-staining because of its radioactivity and the possible risk of cancer. } We are wondered about the relative 'light' safety instructions for handling } and disposal uranylsolutions compared to that for C-Isotopes used in spe- } cial enzymatic reactions. } 1.Is this risk negligible ,
A qualified yes. The amount of activity is very low, and U is an alpha-particle emitter. The range of the alphas is less than the thickness of the dead layer of skin, so UAc is not a radiation hazard if one gets it on ones hands, etc. However, alpha emitters can be extremely hazardous if they are inhaled or ingested, so precautions should be taken so that small droplets are not produced, and always wash your hands after using UAc.
} if not, what intern al safety instructions exists in other labs or is } literature about the risk of uranylacetat in biological labs available?
Your safety office should have info on internal procedures; these can vary from place to place. In the US we can get a materials safety data sheet (MSDS) for any substance, and this will give info about hazards, re- strictions for transport and use, etc. I'm sure any of the local EM sup- pliers can get this for you. Yours, Bill Tivol
Kristof Kovacs wrote: ================================================ I am going to be transiting in Miami on my way to Cancun and have been told that I would need a VISA for these few minutes in the United States. Is that really true? ================================================= There seems to be a lot of ambiguity about this and if you call the same airline three times, you can get three different opinions on this. I can understand how there could have been some confusion on this.
We are talking about someone travelling to Cancun, transiting some point in the USA en route to Cancun, and they would be travelling on a passport for which a visa for travel to the USA would normally be required. Anyone else has no worries whatsoever.
I called United Airlines three times and received three different answers. I called Lufthansa twice and got still other answers. But this is what seems to be the correct situation:
If a passenger (as described above) arrives in the USA as a transit passenger, if they are holding a confirmed onward ticket for entry into a third country, you are exempted from the visa requirement and you do not need a visa. There is one exception to this and that is, if the transit time is more than eight hours. Then you lose your eligibility for the exemption and you would need a visa (according to "Linda" at LH). Someone "overnighting" at their transit point could fall into this category.
This should not be a cause for concern or worry. If you do have any such concerns, contact the airline bringing you to the USA point of entry, and make sure that in your record they have "documented the record" as to what passport you are travelling on and that you won't need a visa for the purposes of transiting the USA. That could save you any kind of hold up or inconvenience when you check in for your first flight.
Linda told me one other interesting thing: She said that sometimes people travel to Cancun and like it (literally) so much, they stay more than 90 days and then they run into a visa requirement of Mexico!
Hope this clarifies things.
Chuck
PS: Remember I am not an expert on this, that is why if you have any further concerns, contact your carrier to the USA for the "last word" on this. And be sure they document what they tell you in your record!
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} Date: Wed, 12 Aug 1998 11:26:21 -0400 (EDT) } From: Judy Trogadis {judy-at-playfair.utoronto.ca} } To: Microscopy-at-sparc5.microscopy.com } Subject: GFP immuno } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are trying to do immunostaining of newt tissue, which, while alive, was } expressing GFP. Cryosectioning is an option. I heard that prolonged fixation } in paraformaldehyde will quench the GFP signal - is that true? The decision } to make is whether to prefix the tissue for longer period of time and risk } losing the signal or quick freeze, section and then fix for shorter period } of time. Also, what would be an optimal fixative for GFP retention? } } Sharing any thoughts or direct experience in this area would be highly } appreciated. } } } Judy Trogadis } Eye Research Institute and } University of Toronto } Toronto Hospital, Western Div. } 399 Bathurst St. } Toronto, Canada M5T 2S8 } } phone: 416-603-5088 } Fax: 416-603-5126 } email: judy-at-playfair.utoronto.ca
We do not have trouble with p-form fixed GPF tissue. What do you call "prolonged."} } If you want to look at ultrastructure, you CANNOT freeze and then fix; you must fix first. I need to know more about what you want to do before commenting on methods. S}
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We have got lots of historical Journals available for those, who are interested in getting a set or filling some shortages in his or his departements library. Please contact me, I will forward your inquiry to Prof. Hildebrand. Please be aware that the delivery charge for shipment has to be payed by the recipient.
Please find attached the counts of the existing volumes.
Can anyone tell me for certain how much BDMA is equivalent(volume to volume) to DMP-30? There are very confusing substitution rules to be found in the literature. Can anyone guarantee the same result if we suddenly switch to BDMA? We really don not have time to run tests. DMP-30 is problematic with its capability of binding water. We use it only for a short time, then discard the rest. But, one never knows how long the reagent has been sitting on a shelf before purchase! Thanks, Hildy Crowley {hcrowley-at-du.edu}
Dear colleagues, I would like to find out if there is a non- fluorescent dye that can be used on fluorescently labeled sections (to identify structures) without compromising the integrity of the fluorescence? Thank you, Lilith --------------------------------------- Lilith Ohannessian-Barry NRC, IBS, Ottawa, Ont. K1A 0R6 Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
} Hi, } } Can anyone tell me for certain how much BDMA is equivalent(volume to } volume) to DMP-30? There are very confusing substitution rules to be } found in the literature. Can anyone guarantee the same result if we } suddenly switch to BDMA? We really don not have time to run tests. } DMP-30 is problematic with its capability of binding water. We use } it only for a short time, then discard the rest. But, one never knows how long the reagent has been sitting on a shelf before purchase! } Thanks, } Hildy Crowley } {hcrowley-at-du.edu}
Hildy:
I have used BDMA in my "Epon" mixes for about 2 years, absolutely no problems. I use 2.5% to 3% BDMA as recommended by Electron Microcopy Sciences in the kit they supply. I do everything by weight. I replace the BDMA 6-8 months after opening, just to be safe and it is stored at room temp.
Geoff
-- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Would any interested Third Party Maintenance organizations drop me an e-mail including their location, area of expertise, main contact, etc. I wish to form a clearinghouse for all TPMs to share technical information, customer referral, etc.
I have been in business as an SEM third party maintenance organization in Southern California for over 15 years. Quite often I am asked to recommend someone or repair equipment outside our geographical area.
We have everything to gain. The ultimate winner is the customer.
I have obtained a Vickers Shearing Eyepiece. It fits in place of an eyepiece in a microscope and apparently through a set of prisms and colored filters it forms two images (one red and one green) that can be separated by means of a micrometer dial in the eyepiece.
Does anyone know what this was used for. My best guess is that it may have been used for measuring particle sizes by "separating" the two images from each other and then reading the micrometer scale.
Dear all, I have had a student from another department come to see me needing some help for his project. He needs a "method for the preparation and analytical procedure to determine algal metal uptake on cell surface and in the various subcellular regions". If anyone has any ideas I would appreciate it very much. TIA Joan Clark
Would any interested Third Party Maintenance organizations drop me an e-mail including their location, area of expertise, main contact, etc. I
wish to form a clearinghouse for all TPMs to share technical information, customer referral, etc.
I have been in business as an SEM third party maintenance organization in Southern California for over 15 years. Quite often I am asked to recommend someone or repair equipment outside our geographical area.
We have everything to gain. The ultimate winner is the customer.
} There seems to be a lot of ambiguity about this and if you call the same } airline three times, you can get three different opinions on this. I can } understand how there could have been some confusion on this.
This is true, mainly because there are different rules for nationals of different countries. When this question arose a couple of days ago my travel agent provided me with a copy of the regulations (about 500 words) applying only to persons travelling in transit through the US!!
If anyone is interested I can look up details from this document, otherwise this information is available from any US Embassy or Consulate or most airlines and travel agents.
Robin
} We are talking about someone travelling to Cancun, transiting some point in } the USA en route to Cancun, and they would be travelling on a passport for } which a visa for travel to the USA would normally be required. Anyone else } has no worries whatsoever. } } I called United Airlines three times and received three different answers. } I called Lufthansa twice and got still other answers. But this is what } seems to be the correct situation: } } If a passenger (as described above) arrives in the USA as a transit } passenger, if they are holding a confirmed onward ticket for entry into a } third country, you are exempted from the visa requirement and you do not } need a visa. There is one exception to this and that is, if the transit } time is more than eight hours. Then you lose your eligibility for the } exemption and you would need a visa (according to "Linda" at LH). Someone } "overnighting" at their transit point could fall into this category. } } This should not be a cause for concern or worry. If you do have any such } concerns, contact the airline bringing you to the USA point of entry, and } make sure that in your record they have "documented the record" as to what } passport you are travelling on and that you won't need a visa for the } purposes of transiting the USA. That could save you any kind of hold up or } inconvenience when you check in for your first flight. } } Linda told me one other interesting thing: She said that sometimes people } travel to Cancun and like it (literally) so much, they stay more than 90 } days and then they run into a visa requirement of Mexico! } } Hope this clarifies things. } } Chuck } } PS: Remember I am not an expert on this, that is why if you have any } further concerns, contact your carrier to the USA for the "last word" on } this. And be sure they document what they tell you in your record! } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== }
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
I have had experience with animal tissues. Presumably, you are contemplating x-ray microanalysis. I would avoid all chemical fixatives !! They cause leaching of soluble ions etc. The specimen needs to be cryofixed in some way (even rapid freezing on a cold copper plate cooled by liquid nitrogen). Then, freeze dried (even by putting them in a vacuum coating unit on a cold copper plate (on plastic insulation to prevent fast warm-up), and allowing to pump for a couple of days. Then sections? Infiltrate in liquid resin under vacuum to assist penetration, take out, change and cure as normal. Then cut 0.5 micron sections on a DRY glass knife, put onto film-coated grids (I use colloidion for ease of preparing them) which may be pre-coated with carbon. Then carbon coat (against charging in STEM). The grid material may be copper, aluminium, titanium, nickel - they may all fluoresce and give false results so be careful!
Hope this gives some idea - it can be done very usefully this way but the ultrastructure will probably be ***p! Freezing damage will be quite extensive but the goodies will all be in there. With freeze drying, some of the crystal damage will be masked because I believe some membranes will 'relax' before being caught in position by the hardening of the resin.
Another approach is to freeze (as above) and then freeze substitute for about 10 days in pure acetone at -80. This can be done simply by suspending the specimens in a container above liquid nitrogen. I use a one-third-full 25 litre dewar with the specimens about 2 inches from the top. You must use a thermocouple or somehow determine this temp. I have a wire tc to a digital thermometer which sits in the base of a brass gauze basket containing the specimens. Then bring to room temperature and infiltrate with resin etc. This method was really meant fr low temperature embedding (which I don't have).
Hope this helps - Keith Ryan Plymouth Marine Lab., UK
} Does anyone know what this was used for. My best guess is that it may have } been used for measuring particle sizes by "separating" the two images from } each other and then reading the micrometer scale.
Exactly. As I remember, you displace one image until it is tangential to the other and read off the diameter (displacement).
I am a designer in Dallas and am doing a poster for a trade show for one of out clients and need to purchase a color scan of dust, spores, mites etc.. to show what type of organisms live in the air we breathe contact me via e-mail and we can discuss cost and availability. my address is jharskjold-at-gpcreative.com.
Hi All: I am looking for a sure-fire method to remove the "speckled" appearance in silicon HREM images. I've heard of etchants and other methods to remove this artifact, but I am unaware of any specific sure-fire recipes/methods. Any contributions would be greatly appreciated. Thanks in advance.
Regards,
Michael Coviello UT Arlington Materials Science Arlington, TX 817 272-5496
Joan, I have done extensive EM on the uptake of metals by different = species of algae and bacteria under experimentally controlled = conditions.. In these cases the algae were incubated with different = concentrations of selected heavy metals. Under these conditions the = metals were adsorbed/ppt to the cell walls of living algae or bacteria = and were clearly visible as discrete particles doing conventional TEM = without contrast enhancement. Under these controlled conditions in which = the metal compounds are added to the media it may not be necessary to do = edx unless you want to do semiquanitative analysis although the = differences will be clearly visible. When the organisms are dying or = already dead, uptake is intracellular/internal. Prepare the samples = with or without osmium postfixation. Postfixation in somecases augments = the visibility depending on the metals. This is a good start and will = reveal a lot of info. It can get complicated and expensive if you want = to go farther with it. Obviously, uptake of metals which are unbound will be loss by this = procedure in which case quenching in a LN2/propane or isopentane = followed by freezesubstitution or freeze drying and embedding or = cryosectioning, etc. is necessary. Hank Adams IMC Cell Biology Baylor College of Medicine
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} Joan, I have done extensive EM on = the uptake of=20 metals by different species of algae and bacteria under experimentally=20 controlled conditions.. In these cases the algae were incubated with = different=20 concentrations of selected heavy metals. Under these conditions = the metals=20 were adsorbed/ppt to the cell walls of living algae or bacteria and were = clearly=20 visible as discrete particles doing conventional TEM without contrast=20 enhancement. Under these controlled conditions in which the metal = compounds are=20 added to the media it may not be necessary to do edx unless you want to = do=20 semiquanitative analysis although the differences will be clearly = visible.=20 {/FONT} {FONT color=3D#000000 size=3D2} When the organisms are dying or = already=20 dead, uptake is intracellular/internal. Prepare the samples = with or=20 without osmium postfixation. {/FONT} {FONT size=3D2} Postfixation in = somecases=20 augments the visibility depending on the metals. This is a good start = and will=20 reveal a lot of info. It can get complicated and expensive if you = want to=20 go farther with it. {/FONT} {/DIV} {DIV} {FONT size=3D2} Obviously, uptake of metals which are unbound = will be=20 loss by this procedure in which case quenching in a LN2/propane or = isopentane=20 followed by freezesubstitution or freeze drying and embedding or = cryosectioning,=20 etc. is necessary. {/FONT} {/DIV} {DIV} {FONT size=3D2} Hank Adams {/FONT} {/DIV} {DIV} {FONT size=3D2} IMC Cell Biology {/FONT} {/DIV} {DIV} {FONT size=3D2} Baylor College of = Medicine {/FONT} {/DIV} {/BODY} {/HTML}
I have located in the archives instructions/schematics for a Kinney SC 3 evaporator. The machine has long since been cannibalized. Anyone wanting this material is welcome to it--otherwise it goes into "file 13" shortly.
Just email with address if interested.
S
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
The Vickers Shearing Eyepieces were well known for their high accuracy and precision in measuring particle sizes. I had the privilege of using them while on an RMS short course in the UK 20 years ago. You are right about the image separation. While I really need to dig out the instructions to be absolutely sure of they work, I remember the following: That you superimposed the images and either set the vernier to zero or read the vernier setting. Next, you rotated the drum until the two images were separate but just touching and read again. It may be that you need to divide the results by 2. Try it with something of known diameter to test. I will rummage through the extensive MME archives and see if I can find more complete instructions. In the meantime, enjoy your new found treasure!
Best regards,
At 10:52 PM 8/13/98 EDT, MICROFAB-at-aol.com wrote:
Hi All: I am looking for a sure-fire method to remove the "speckled" appearance in silicon HREM images. I've heard of etchants and other methods to remove this artifact, but I am unaware of any specific sure-fire recipes/methods. Any contributions would be greatly appreciated. Thanks in advance.
Regards,
Michael Coviello UT Arlington Materials Science Arlington, TX 817 272-5496
Does anyone have a Nikon 40X/NA 1.2 water immersion objective for sale? Any leads would be appreciated.
Most Nikon water immersion lenses for 160 mm tube models are out of stock.
Thanks, Glen
-- Glen MacDonald Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax The box said "Requires Windows95 or better". So I bought a Macintosh.
What is the source of the "speckle" to which you are referring? If it is shot noise, no etchant will have any effect. If the images have a hazy appearance due to the presence of a damaged surface layer (produced by ion milling?), you might try an alternate specimen preparation route such as chemical polishing.
++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
I suggest getting in touch with companies that make air filters (3M, for example) or your local eye/ear/nose/throat specialist. How about David Scharf?
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
I can't thank you enough for the rapid outpouring of responses to my request. With your help, I was able to generate a list of about twenty labs for my grateful customer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
Shot noise refers to poor statistical sampling in an image. For instance if N electrons arrive per pixel of the image, this will lead to a random component in the "counting" of the electrons which is of the order of root(N). Most textbooks on HREM will have a discussion of this. It is why one does not use the fastest possible film (and short exposure times) in recording HREM images, which could otherwise be done to avoid problems with drift. -Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
Speckle in HREM images? How high was the mag? Are you sure you aren't looking at the considerable ion milling radiation damage artifact that forms in Si even after only a few seconds of ion milling? I suppose the mag would have to be around 250 to 400kx, which isn't very HREM, for this radiation damage to be confused with shot noise/speckle. At lower mags the radiation damage looks like salt and pepper.
5 to 10 nm is exactly the size of the ion mill, artifactual, radiation damage, salt-and-pepper.
We try to thin specimens via tripod polishing so we don't have to ion mill for this reason. We are successful better than half the time. When we do ion mill, we do so at 2 to 3 KeV for no more than one or two minutes. We note that even 15 to 30 seconds of this low dose ion mill will create ion mill artifact. If it bothers you, don't ion mill. But, about 99% of people publishing TEMs of ion milled Si don't mention it--even those doing radiation damage studies. (OK, so I exaggerate a little).
The other responders did an excellent job of explaining shot noise, etc.
Coviello-at-mae.uta.edu on 08/14/98 03:37:15 PM Please respond to Coviello-at-mae.uta.edu To: Ronald Anderson/Fishkill/IBM-at-ibmus cc:
Hi,
Recently there has been some interest in Araldite 502 as an embedding medium. In our laboratory we have been using the mixed resin embedding which appeared in 1972 in Hayat's book "Basic Electron Microscopy Techniques". Rather than use it as a stock mixture I recalculated it to be a single mixture. (Araldite 502 30ml, Eponate 12 or LX-112 50ml, DDSA 110ml, DMP-30 1.5%, dibutyl phthalate 1/2%-2%.
Dibutyl is a plasticicer for epoxides. It does not react chemically with the monomers, but it allows for slippage along the sites of crosslinks. This influences the hardness and cuttability of the block. With a standardized 48 hour, 60deg C. polymerization time, using 1/2% dibutyl yields medium hard block, 1% gives a medium soft block, and 2% a very soft block.
The medium hard block is very good for immediate thin sectioning. The medium soft block (after 24 hrs only of polymerization at 60deg C.) is ideal for post-embedding immunogold for TEM due to its low crosslinkage. The soft block is great for endless thick sections. Glass knives last a long time, probably due to the absence of NMA. As a matter of fact, the above formulation with 1% dibutyl is the only formulation which should be employed when no diamond knives are available for thin sectioning. Araldite 502 already contains a large amount of dibutyl when purchased. That such a small variation in additions of dibutyl should make a difference in sectioning qualities has always been a mystery to me. Sometimes, an attempt is made to substitue one of the other Araldites, such as 506 for 502. The Araldites are not interchangeable. There are vast differences in composition between labels.
Any of the above mixtures can be repolymerized at 95deg C. until a block firmness of desirable qualities is achieved.
The big disadvantage of this mixture is its viscosity. Infiltration times must be lengthened over times used with epon. The mixture seperates easily and therefore must be kept well agitated. Infiltration must take place on a rotator, not on a rocker. The above formulations are unsuitable for reembedding thick sections from microscope slides. Araldite is known for its elasticity and its adhesive qualities in industry - this is counterproductive to clean seperations of sections and blocks on slides.
I have used the above formulations since 1981 for students (easy sectioning), for projects which require huge amounts of thick sections, and for immunocytochemistry-postembedding Au. They are a good tool in the laboratory. Bye, Hildy
Ken Tobin Guyot Hall Dept. of Geosciences Princeton University Princeton, NJ 08544 Ph: 609-258-1383 FAX: 609-258-1274 e-mail: tobin-at-geo.princeton.edu
I have noted with interest the several recent postings regarding the Tracor Northern ADEM SEM, its motivation and demise. Tim Moeller's postings covered the history of this development quite well, but as one who was at the center of both the development efforts and also the management decisions leading up to it, the following remarks from my perspective may be of interest.
It is generally taken as a "given" that our principal failure was that we failed to anticipate the impact on Tracor Northern EDS sales due to the reaction of the other SEM manufacturers to our entry into their market -- the so called "channel conflict". We were, in fact, deathly afraid of this scenario and I remember numerous conversations with various persons through the 70's and early 80's where I argued why it would be sheer folly for us to attempt to build our own SEM (an idea which numerous people suggested to us). So why did I change my mind, champion the effort, and ultimately give up my position as head of R&D at TN in order to lead the ADEM development team?
The decision to proceed with ADEM took place in 1983, shortly after TN's launch of the very successful TN-5500 analyzer (though the major development effort didn't kick in till spring of 1985).. Though we were enjoying great market success at that time, we saw a disconcerting future on the horizon. At that time a very large fraction of our sales were based on automating other people's microscopes -- large systems involving digital imaging and SEM automation (motorized stages, column control, etc.), together with the application software to operate them. It was commonplace to sell expensive and complex systems of which perhaps 1/3 or less of TN's revenue was due to the EDS components as such. It was in the "computerized microscope" arena where we felt that we had established a technological edge in the market. However, we perceived that this was an unstable situation and that it was only going to be a matter of time before the SEM manufacturers incorporated imaging and automation capabilities into their systems. When this occurred, we reasoned, the EDS market was going to be reduced to one of selling commodity EDS components. Rather than passively await this fate, we decided to be bold and enter the SEM market ourselves, since that is where we thought the real long-term action and opportunity lay. Though we were very anxious about the impact on our EDS sales, we felt that we were at a "window of opportunity" where we could pull this off, but that the window was sure to close. With this in mind, we decided to make our move. The final go-ahead was made with the full understanding of 'Van' Vandenheuvel (general manager of TN), Bill Buffo (president of TN and Tracor VP of the Instruments Group) and Frank McBee (president of Tracor) that this was going to have a negative impact on EDS sales, but it was felt that we could weather this as Tracor was a very healthy company at that point. Since TN had been a consistently profitable company for the past 20 years, it was Tracor's position that it was appropriate to invest short-term profits for a future position in a market where we thought we had something unique to offer.
Central to our strategy was a conviction that we could provide a substantially better value by designing a fully integrated SEM and thus offer both better price and productivity than was possible by the "piecemeal" approach of installing an EDS/automation system on a stand-alone SEM manufactured by someone else. I think we actually delivered on this rather well. Despite the fact that ADEM was indeed a rather complicated system, to be fair you need to compare it with the "glued together" systems it was designed to replace. Compared to these, ADEM was a bargain, simple to run and maintain, and highly productive. I am told that the instruments in the field have proven to be reliable, and had the development and support team remained intact, I suspect the instrument could have enjoyed a long life. So what went wrong? Having thought about it a lot in the subsequent years, I note three key "surprises" which ultimately played a large role in dooming our efforts:
(1) Introduction of field-emission SEM technology; (2) The leveraged buyout of Tracor; and (3) The outbreak of "global peace".
Field emission was a technology breakthrough we didn't anticipate. We were able to keep ADEM under wraps until its market introduction in the summer of 1987, and thus stave off erosion of EDS sales until we were ready to make our move. The introduction was quite successful, and those present at the Baltimore EMSA/MAS introduction may remember the SRO crowds it drew. There was no question that ADEM made a big splash. However, it was around this time that Hitachi introduced their field-emission SEM and this quickly became the technology where people with ADEM-sized budgets thought they should spend their SEM dollars -- particularly so in the semiconductor industry, which by now had evolved into being the principal market for high-end SEMs. We had targeted the existing thermionic SEMs of the early 80's, and felt that we competed quite well with the instruments of that type -- but field emission was a "paradigm shift" for which we were unprepared. However, we were playing for the "long term" and our development team knew we would have to work both hard and smart to pull off what we were attempting. We started work on our own field emission instrument.
About this time, the financial bottom was also dropping out of Tracor. With incredible ill-timing, a private investment group had finalized a leveraged buyout of Tracor just days before the major market crash of the late '80s. Then, the collapse of the Soviet Union depressed the defense industry on which Tracor relied for much of their revenue. Also around this time, McBee, Buffo, and Van all resigned, and with them went the commitment to the product. Instead of struggling through a "trough" as our SEM sales replaced lost EDS sales, as we expected, we found the whole company fighting for survival. Eventually, Tracor was forced to sell off the instruments group and the new management decided that they had to jettison ADEM in late 1990 -- that's when I joined Rich Lee to participate in his vision of a low-cost automatable SEM.
Could ADEM have succeeded had these "surprises" not taken place? I like to think so, but I also now see the huge obstacles we faced with a clarity that only hindsight can provide. I think there was a certain amount of "hubris" involved in our failure, and I accept the responsibility for that. There were also some engineering decisions I/we made which are questionable in retrospect. ADEM was loaded with technological innovations, but I do wish in retrospect that I had been more selective and focused -- had we kept the product simpler (and less expensive) I think we might have vastly improved our chances. (That's a hard-learned lesson that I am happy to say that we have been able to apply to the development of the PERSONAL SEM at RJ Lee Instruments -- and with a lot more satisfactory business outcome, I might add). And, of course, had we had access to the kind of powerful/inexpensive PC technology which is available today, we could have saved a whole lot of engineering and provided a lot more bang for the buck -- another lesson learned.
The whole ADEM experience did leave me with deeply conflicted feelings. On the one hand, I remain very proud of what our ADEM team accomplished, and that we had the nerve to take on what we knew was a huge challenge. But at the same time I am also painfully aware of the personal and economic costs and the "bottom line" fact that we ultimately failed to achieve our vision. The one thing I can say with certainty is that the ADEM team was spectacularly competent and, based on their dedication and effort, they deserved an outcome far better than what I was able to deliver.
A final bit of irony -- while we were laboring in our secret quarters in an industrial park miles from the main TN plant, the ADEM team occasionally noted the weird activities of a nearby startup. It turned out they were marketing collector dolls. Fast-forward seven years. I have an article on my desk from our local Pittsburgh paper describing the phenomenon that the Pleasant Company "American Girl" dolls have become -- stating that the company turned a PROFIT of $80 million last year! Mattel recently purchased the company for something north of $300 million, as I recall. ADEM has been a defunct product for years. Is there a lesson here?
Fred Schamber RJ Lee Instruments Limited ....................... mailto:fhscham-at-SGI.NET
After ~ 2 weeks of testing I am going to test the full filtering of Email to the ListServer this weekend. Any mail which triggers a Filter Flag is tagged as potential JUNK/SPAM mail and WILL NOT be posted to the ListServer. The sender of the suspect Email is sent a warning message. If you receive one of these warning messages please follow the instructions you receive so that we can sort out any remaining issues.
Remember the filter only operates on the text in the Email Header and Subject lines. It does not search the body of the text . Suspect or Bogus Email Addresses, Keywords in the Subject line (or lack of a Subject) are the some of criteria I am using.
} From the subscriber base the only problem that I expect will be the occasional rejection due to suspect Email address, this will be true if your local Email provider changes and/or modifies headers in your message which makes your mail appear to be posted from a different address. When this happens just follow the instructions and send me a confirming message, I can then add your particuliar Email address to a list of "exceptions" and your postings will go through by-passing the filter.
There are obviously no guarentee's that all junk mail will be filtered, but this should handle the majority (I hope).
If things go well, and there are no major problems over the weekend (when the mail is "light"), then I will obviously leave the filter on-line. Please report problems to me at:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have noted with interest the several recent postings regarding the } Tracor Northern ADEM SEM, its motivation and demise. Tim Moeller's } postings covered the history of this development quite well, but as one } who was at the center of both the development efforts and also the } management decisions leading up to it, the following remarks from my } perspective may be of interest. } } It is generally taken as a "given" that our principal failure was that } we failed to anticipate the impact on Tracor Northern EDS sales due to } the reaction of the other SEM manufacturers to our entry into their } market -- the so called "channel conflict". We were, in fact, deathly } afraid of this scenario and I remember numerous conversations with } various persons through the 70's and early 80's where I argued why it } would be sheer folly for us to attempt to build our own SEM (an idea } which numerous people suggested to us). So why did I change my mind, } champion the effort, and ultimately give up my position as head of R&D } at TN in order to lead the ADEM development team? } } The decision to proceed with ADEM took place in 1983, shortly after TN's } launch of the very successful TN-5500 analyzer (though the major } development effort didn't kick in till spring of 1985).. Though we were } enjoying great market success at that time, we saw a disconcerting } future on the horizon. At that time a very large fraction of our sales } were based on automating other people's microscopes -- large systems } involving digital imaging and SEM automation (motorized stages, column } control, etc.), together with the application software to operate them. } It was commonplace to sell expensive and complex systems of which } perhaps 1/3 or less of TN's revenue was due to the EDS components as } such. It was in the "computerized microscope" arena where we felt that } we had established a technological edge in the market. However, we } perceived that this was an unstable situation and that it was only going } to be a matter of time before the SEM manufacturers incorporated imaging } and automation capabilities into their systems. When this occurred, we } reasoned, the EDS market was going to be reduced to one of selling } commodity EDS components. Rather than passively await this fate, we } decided to be bold and enter the SEM market ourselves, since that is } where we thought the real long-term action and opportunity lay. Though } we were very anxious about the impact on our EDS sales, we felt that we } were at a "window of opportunity" where we could pull this off, but that } the window was sure to close. With this in mind, we decided to make } our move. The final go-ahead was made with the full understanding of } 'Van' Vandenheuvel (general manager of TN), Bill Buffo (president of TN } and Tracor VP of the Instruments Group) and Frank McBee (president of } Tracor) that this was going to have a negative impact on EDS sales, but } it was felt that we could weather this as Tracor was a very healthy } company at that point. Since TN had been a consistently profitable } company for the past 20 years, it was Tracor's position that it was } appropriate to invest short-term profits for a future position in a } market where we thought we had something unique to offer. } } Central to our strategy was a conviction that we could provide a } substantially better value by designing a fully integrated SEM and thus } offer both better price and productivity than was possible by the } "piecemeal" approach of installing an EDS/automation system on a } stand-alone SEM manufactured by someone else. I think we actually } delivered on this rather well. Despite the fact that ADEM was indeed a } rather complicated system, to be fair you need to compare it with the } "glued together" systems it was designed to replace. Compared to these, } ADEM was a bargain, simple to run and maintain, and highly productive. I } am told that the instruments in the field have proven to be reliable, } and had the development and support team remained intact, I suspect the } instrument could have enjoyed a long life. So what went wrong? Having } thought about it a lot in the subsequent years, I note three key } "surprises" which ultimately played a large role in dooming our efforts: } } (1) Introduction of field-emission SEM technology; } (2) The leveraged buyout of Tracor; and } (3) The outbreak of "global peace". } } Field emission was a technology breakthrough we didn't anticipate. We } were able to keep ADEM under wraps until its market introduction in the } summer of 1987, and thus stave off erosion of EDS sales until we were } ready to make our move. The introduction was quite successful, and } those present at the Baltimore EMSA/MAS introduction may remember the } SRO crowds it drew. There was no question that ADEM made a big splash. } However, it was around this time that Hitachi introduced their } field-emission SEM and this quickly became the technology where people } with ADEM-sized budgets thought they should spend their SEM dollars -- } particularly so in the semiconductor industry, which by now had evolved } into being the principal market for high-end SEMs. We had targeted the } existing thermionic SEMs of the early 80's, and felt that we competed } quite well with the instruments of that type -- but field emission was a } "paradigm shift" for which we were unprepared. However, we were playing } for the "long term" and our development team knew we would have to work } both hard and smart to pull off what we were attempting. We started } work on our own field emission instrument. } } About this time, the financial bottom was also dropping out of Tracor. } With incredible ill-timing, a private investment group had finalized a } leveraged buyout of Tracor just days before the major market crash of } the late '80s. Then, the collapse of the Soviet Union depressed the } defense industry on which Tracor relied for much of their revenue. Also } around this time, McBee, Buffo, and Van all resigned, and with them went } the commitment to the product. Instead of struggling through a "trough" } as our SEM sales replaced lost EDS sales, as we expected, we found the } whole company fighting for survival. Eventually, Tracor was forced to } sell off the instruments group and the new management decided that they } had to jettison ADEM in late 1990 -- that's when I joined Rich Lee to } participate in his vision of a low-cost automatable SEM. } } Could ADEM have succeeded had these "surprises" not taken place? I like } to think so, but I also now see the huge obstacles we faced with a } clarity that only hindsight can provide. I think there was a certain } amount of "hubris" involved in our failure, and I accept the } responsibility for that. There were also some engineering decisions I/we } made which are questionable in retrospect. ADEM was loaded with } technological innovations, but I do wish in retrospect that I had been } more selective and focused -- had we kept the product simpler (and less } expensive) I think we might have vastly improved our chances. (That's a } hard-learned lesson that I am happy to say that we have been able to } apply to the development of the PERSONAL SEM at RJ Lee Instruments -- } and with a lot more satisfactory business outcome, I might add). And, } of course, had we had access to the kind of powerful/inexpensive PC } technology which is available today, we could have saved a whole lot of } engineering and provided a lot more bang for the buck -- another lesson } learned. } } The whole ADEM experience did leave me with deeply conflicted feelings. } On the one hand, I remain very proud of what our ADEM team accomplished, } and that we had the nerve to take on what we knew was a huge challenge. } But at the same time I am also painfully aware of the personal and } economic costs and the "bottom line" fact that we ultimately failed to } achieve our vision. The one thing I can say with certainty is that the } ADEM team was spectacularly competent and, based on their dedication and } effort, they deserved an outcome far better than what I was able to } deliver. } } A final bit of irony -- while we were laboring in our secret quarters in } an industrial park miles from the main TN plant, the ADEM team } occasionally noted the weird activities of a nearby startup. It turned } out they were marketing collector dolls. Fast-forward seven years. I } have an article on my desk from our local Pittsburgh paper describing } the phenomenon that the Pleasant Company "American Girl" dolls have } become -- stating that the company turned a PROFIT of $80 million last } year! Mattel recently purchased the company for something north of $300 } million, as I recall. ADEM has been a defunct product for years. Is } there a lesson here? } } Fred Schamber } RJ Lee Instruments Limited } ....................... } mailto:fhscham-at-SGI.NET
Leitz, Model SM-LUX, binocular head with 10x Periplan eyepieces. The 170mm tube length objectives are: 3.5/0.10, 10/0.25, 45/0.65, and 100/1.30 oil, the 45x and 100x are spring loaded noses. All the objectives lens are Leitz. The microscope has a light source with transformer. Everything works smoothly and the optics are crisp and clear. There is a small wear spot on the frame where the paint has been rubbed away by the support inside the case. Otherwise it is in very good condition.
The sequence of ADEM postings tell a fascinating story that deserves more than the obscurity of the listserver archives. Will you consider publishing? [Are you reading this, Don Grimes?]
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
This is not a SM-LUX but a SM microscope and worth to my knowledge not more than $300.
Joseph Passero wrote:
} Leitz, Model SM-LUX, binocular head with 10x Periplan eyepieces. The } 170mm tube length objectives are: 3.5/0.10, 10/0.25, 45/0.65, and } 100/1.30 oil, the 45x and 100x are spring loaded noses. All the } objectives lens are Leitz. The microscope has a light source with } transformer. Everything works smoothly and the optics are crisp and } clear. There is a small wear spot on the frame where the paint has been } rubbed away by the support inside the case. Otherwise it is in very good } condition. } } See attached photo file. } } Entertaining Offers } } Thank You } } Joseph Passero } jp-at-spacelab.net } } ------------------------------------------------------------------------ } [Image]
Does anyone have comments on the suitability of this printer for TEM working prints? The input would be from a 1024x1024 digital camera and a SprintScan 45 from large-format negatives. A comparison with the Epson Stylus would be useful. Thanks Sally
---------------------------------------------------------------------- Dr Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: GPO Box 475 ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 (0)2 6249 2743 |Australian National Univ. FAX 61 (0)2 6249 4891 |Canberra, Australia 2601 http://online.anu.edu.au/EMU/home.htm
As a member of the ADEM development/manufacturing/service team, I would like to respond to the recent postings on this subject. As a member of Fred Schamber's team at Tracor, I would just like to say that I am grateful and honored to have had the opportunity to work with such an intense, innovative, and fun group of people. I joined the ADEM project just months before its introduction and worked as a final assembly and test technician. The knowledge I gained from that experience is inmeasurable.
In my opinion, there is no way the demise of the ADEM can be attributed to any decision or action that Fred made. There were so many external influences at play at the time that I think we never really had a chance to meet the "bottom line" in the time frame the corporation wanted. I think I speak for all the members of the ADEM team when, if asked if I'd do it all again I can only say this: In a heartbeat!
We keep our DMP-30 tightly sealed in the refrigerator and bring it to room temperature (20 deg C) in the fume hood for the 3 days of resin infiltration and embedding. Always keep lid closed when not in use, use a clean disposable glass pipette each time. Once finished store back in the refrigerator. Have stored DMP-30 for up to 2 years - discard if it appears too yellow or becomes slightly granular.
Haven't tried BDMA as have had no problems with DMP-30
Belinda White Senior Technician Centre for Electron Microscopy University of Natal Scottsville,Pietermaritzburg South Africa
If the contrast is from ion milling as Ron Anderson suggests, you might want to try the small angle cleavage technique which will eliminate this as well as the ion mixing/amorphization layers on these samples. In fact, one of the disadvantages of the technique is a lack of amorphous areas for focussing. Silicon feathers down to almost nothing and will provide you with extremely thin areas. You can produce XTEM and plan view samples using this technique. If you need site-specific preparation, this technique will not work for you. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Mike Coviello To: microscopy-at-sparc5.microscopy.com
Hi All: I am looking for a sure-fire method to remove the "speckled" appearance in silicon HREM images. I've heard of etchants and other methods to remove this artifact, but I am unaware of any specific sure-fire recipes/methods. Any contributions would be greatly appreciated. Thanks in advance.
Regards,
Michael Coviello UT Arlington Materials Science Arlington, TX 817 272-5496
Belinda White wrote: ================================================ We keep our DMP-30 tightly sealed in the refrigerator and bring it to room temperature (20 deg C) in the fume hood for the 3 days of resin infiltration and embedding. Always keep lid closed when not in use, use a clean disposable glass pipette each time. Once finished store back in the refrigerator. Have stored DMP-30 for up to 2 years - discard if it appears too yellow or becomes slightly granular.
Haven't tried BDMA as have had no problems with DMP-30 =============================================== One reason why DMP-30 has retained its popularity in many markets of the world is that it is not a HAZMAT from the standpoint of shipping. The inclusion of one bottle of BDMA, which is a HAZMAT (UN#2619), into an embedding resin kit, which otherwise does not contain any other HAZMATs, increases greatly the shipping costs, since it can not go by cheaper methods and can go only by air freight (or sea freight). Yet as was pointed out, many people get laboratory results that are just fine with DMP-30.
On the other hand, and it is not widely known, by taking advantage of certain small quantity shipping exemptions, one can still get their BDMA in such markets relatively inexpensively, but by using a different packaging approach, and this approach is explained on our website under "Hazardous Materials: Good News and Bad News". For the US domestic market, these differences are very small and are not significant. The bigger differences come into play when making shipments over international boundaries.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
As most regular subscribers know, Microscopic Explorations, a hands on science program designed to teach microscopy has recently been publishe= d by the Lawrence Hall of Science. The book is a direct result of Project M= icro and the monumental efforts of Caroline Schooley. It has many interesti= ng microscopic activities for 9-12 year old students including one activit= y where students examine sand with their microscopes. The Midwest Micros= copy and Microanalysis is working with the local science teachers to put tog= ether several kits to support this program, and we need sand from around the world. If you would like to donate a sample please dry out a handful o= f sand, seal it in a plastic bag (Ziplock will work), and mail it to:
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd Abbott Park, IL 60064-3537
Please include the location the sand was collected from. Your donation= s will be greatly appreciated. If we get enough donations we will be happy to= share them with fellow scientists who are supporting science education.
Please accept my apologies for posting this, as it is not strictly a microscopy-related question, but I was hoping that some of our esteemed subscribers would be able to offer some advice, or point me in the right direction.
We are looking at proliferating cells in animal tissues and, at present, are giving the animals (goats) 100mg/Kg body weight IV two hours prior to sacrifice. One of our researchers has suggested giving the animals another dose approx 5-7 days prior to this one.
Before we consider this, we wish to know a couple of things about BrdU, and have been unable to obtain any information from our supplier (Sigma). How long does BrdU remain in the bloodstream before being excreted, and which organ(s) is responsible for BrdU breakdown and excretion. What is the half-life of BrdU, ie, how long can the BrdU be detected immunocytochemically in cells after initial dosage? What accumulative effect will the BrdU have at this dose if given several times over a short time frame, say three weeks?
} Has anyone out there tried antigen retrieval method on EM samples?
} Qing Yang, Ph.D.
} National Institutes of Health
----------------------------------------
Dear Qing: Two References from my Microwave Methods database that may be of help!
1. Utsunomiya H, Shan L, Kawano I, Iwasaki A, Ono K, Kobayashi A, Kuma K, Kishikawa S, Kakudo K: Immunolocalization of parathyroid hormone in human parathyroid glands with special references to microwave antigen retrieval. Endocr Pathol 1995, 6:223-227
The subcellular localization of parathyroid hormone (PTH) in the normal human parathyroid glands with particular reference to microwave antigen retrieval was investigated using peroxidase-labeled PTH antibody, immunohistochemical, and immunoelectron microscopic methods. The results revealed that PTH granules existed mainly as pro-PTH on the trans side of Golgi and in the regions adjacent to Golgi apparatus. Only a small proportion of secretory granules were stored near the plasma membrane. Microwave irradiation was essential for the immunodetection of PTH. As the irradiative time extended from 1 to 30 min, the staining intensity increased, and the subcellular preservation decreased. Microwave irradiation for 15 min (with the sections in citrate buffer) with a power output of 500 W is the most ideal for PTH antigen retrieval, as well as for subcellular preservation.
2. Stirling JW, Graff PS: Antigen unmasking for immunoelectron microscopy: labeling is improved by treating with sodium ethoxide or sodium metaperiodate, then heating on retrieval medium. J Histochem Cytochem 1995, 43:115-123
To optimize the ultrastructural localization of immunoglobulin G in corneal crystalloid deposits, we compared a range of antigen unmasking techniques. A human corneal biopsy specimen was fixed in formalin, post-osmicated, and embedded in epoxy resin for electron microscopy. Thin sections were immunogold-labeled for IgG after treatment with sodium ethoxide or sodium metaperiodate. Sections were also treated by heating them at 95 degrees C while they floated on water, 0.01 M citrate buffer (pH 6.0), or sodium metaperiodate. The treatments were applied separately and combined. After labeling, crystalloids in untreated sections had a probe density of 5 particles/microns2. Crystalloids in sections treated only with sodium ethoxide or sodium metaperiodate had probe densities of 15-20 particles/microns2. Sodium ethoxide combined with heating on water, or citrate buffer, gave probe densities of 140-160 particles/microns2. Sodium metaperiodate combined with heating on citrate buffer gave the highest probe density (195 particles/microns2). Although sodium ethoxide coupled with heating increased probe density, the ethoxide etched the sections and caused unacceptable damage. Treatment with sodium metaperiodate followed by heating on citrate buffer is recommended for antigen unmasking. This combination gave a high probe density and sections remained intact, with good ultrastructural detail.
FALL 1998 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-V2)
NASSAU COMMUNITY COLLEGE, Long Island, NY
A fourteen week, fall 1998 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered ONE EVENING PER WEEK, Thursdays, starting at 5:30pm. Classes will begin on Sept. 3 and end on Dec. 10, 1998.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 per credit.
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
For those without www access, the catalog description is specified below. If you have further questions, you should e-mail me directly at the address below.
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students. Seats are still available for this section. The college is currently processing late registrations.
Questions regarding the actual registration process can be directed to our registrar at (516) 572-7355. ________________________________________________________________________________
CATALOG DESCRIPTION BIO 221: Transmission Electron Microscopy -- 4 credits Prerequisites: BIO 109-110 or equivalent, CHE 151-152 or equivalent. An introduction to the basic principles of transmission electron microscopy including tissue preparation, microscope (TEM) operation, black & white photography, and micrograph interpretation. The entire laboratory is devoted to the development of skills and preparative techniques involved with the operation of an actual transmission electron microscope. (3 lecture, 3 laboratory hours). Laboratory fee applies. ________________________________________________________________________________
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Are you interested in Digital microscopy? If so, exciting things are = happening at Sound Vision Inc. The founders of the acclaimed Leaf = Systems, the MicroLumina and other digital products have developed the = SVMicro, a new high-resolution digital camera for the microscope for = about $2000.=20
a.. The camera mounts directly to the microscope via a c-mount and = is tethered to your computer.=20 b.. The software allows instant archiving of images in a simple = click and save fashion.=20 c.. The SVMicro utilizes three shot RGB technology that gives you = selectable resolutions for various output file sizes up to 9MB. d.. It is perfect for sending images across the Internet, = documentation, archiving or high-resolution publication.=20 e.. The camera is available with ECP parallel port PC or a SCSI = connection for MAC users. f.. The same camera also takes great black and white pictures in a = one shot mode. g.. Able to do long exposures for most types of fluorescence without = any high cost cooling system h.. Able to rotate the green filter in front of the sensor to take = one shot images=20 i.. Can save sequential shots to your hard drive without leaving the = plugin. To find out more about the SVMicro Digital Camera for the microscope, = please visit our web site at www.soundvisioninc.com. You can also call = Sound Vision=92s sales department at 508-270-0044. Or send me an email = at gwray-at-soundvisioninc.com.=20
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.2106.6"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {B} {FONT size=3D2} {P} Are you interested in Digital microscopy? {/B} If so, exciting things = are=20 happening at {B} Sound Vision Inc {/B} . The founders of the acclaimed Leaf =
Systems, the MicroLumina and other digital products have developed the=20 {B} SVMicro {/B} , a new high-resolution digital camera for the microscope = for=20 about {B} $2000 {/B} . {/P} {UL} {LI} The camera mounts directly to the microscope via a c-mount and = is=20 tethered to your computer. {/LI} {LI} The software allows instant archiving of images in a simple = click and=20 save fashion. {/LI} {LI} The SVMicro utilizes three shot RGB technology that gives you = selectable=20 resolutions for various output file sizes up to 9MB. {/LI} {LI} It is perfect for sending images across the Internet, = documentation,=20 archiving or high-resolution publication. {/LI} {LI} The camera is available with ECP parallel port PC or a SCSI = connection=20 for MAC users. {/LI} {LI} The same camera also takes great black and white pictures in a = one shot=20 mode. {/LI} {LI} Able to do long exposures for most types of fluorescence without = any=20 high cost cooling system {/LI} {LI} Able to rotate the green filter in front of the sensor to take = one shot=20 images {/LI} {LI} Can save sequential shots to your hard drive without leaving the =
plugin. {/LI} {/UL} {P} To find out more about the SVMicro Digital Camera for the microscope, = please=20 visit our web site at {/FONT} {A = href=3D"http://www.soundvisioninc.com/"} {FONT=20 size=3D2} www.soundvisioninc.com {/FONT} {/A} {FONT size=3D2} . You can also = call Sound=20 Vision’s sales department at 508-270-0044. Or send me an email at=20 {/FONT} {A href=3D"mailto:gwray-at-soundvisioninc.com"} {FONT=20 size=3D2} gwray-at-soundvisioninc.com {/FONT} {/A} {U} {FONT size=3D2} . {/P} {/U} {P} {/P} {P} Best Regards, {/P} {P} {/P} {P} Gary Ray {/P} {P} Sales Department {/P} {P} Sound Vision Inc. {/P} {/FONT} {/DIV} {/BODY} {/HTML}
This is a multi-part message in MIME format. --------------C13D3920A222C3F36012EB4A Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Just a personal note of thanks for filling in the gaps, Fred! I agree completely with Caroline Schooley's statement that these posts tell a fascinating story that deserve more than the obscurity of the listserver archives. I would take issue with only one thing you said -- indeed, you take too much upon yourself by saying that the ADEM team "deserved an outcome far better than what I was able to deliver." Of course, anybody who knows your modest and generous nature would almost expect you to say such a thing, accepting responsibility where no real culpability exists, but everyone here should know that you did us no wrong. Certainly it was regrettable, but no one blames you personally for the outcome brought about by factors quite beyond your (or anyone else's) control. To the contrary, I'm sure many team members feel as I do that it was a tremendous privlege to have been included at all, and look back with fond memories on that once-in-a-lifetime experience. We took our best shot, and lost -- but we almost won, and would have won big. That was the chance we took; sometimes you get the bear and sometimes the bear gets you. Besides, there were other personal benefits accrued through the experience, despite the outcome. Anyway, thanks again!
Fred Schamber wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have noted with interest the several recent postings regarding the } Tracor Northern ADEM SEM, its motivation and demise. Tim Moeller's } postings covered the history of this development quite well, but as one } who was at the center of both the development efforts and also the } management decisions leading up to it, the following remarks from my } perspective may be of interest. } } It is generally taken as a "given" that our principal failure was that } we failed to anticipate the impact on Tracor Northern EDS sales due to } the reaction of the other SEM manufacturers to our entry into their } market -- the so called "channel conflict". We were, in fact, deathly } afraid of this scenario and I remember numerous conversations with } various persons through the 70's and early 80's where I argued why it } would be sheer folly for us to attempt to build our own SEM (an idea } which numerous people suggested to us). So why did I change my mind, } champion the effort, and ultimately give up my position as head of R&D } at TN in order to lead the ADEM development team? } } The decision to proceed with ADEM took place in 1983, shortly after TN's } launch of the very successful TN-5500 analyzer (though the major } development effort didn't kick in till spring of 1985).. Though we were } enjoying great market success at that time, we saw a disconcerting } future on the horizon. At that time a very large fraction of our sales } were based on automating other people's microscopes -- large systems } involving digital imaging and SEM automation (motorized stages, column } control, etc.), together with the application software to operate them. } It was commonplace to sell expensive and complex systems of which } perhaps 1/3 or less of TN's revenue was due to the EDS components as } such. It was in the "computerized microscope" arena where we felt that } we had established a technological edge in the market. However, we } perceived that this was an unstable situation and that it was only going } to be a matter of time before the SEM manufacturers incorporated imaging } and automation capabilities into their systems. When this occurred, we } reasoned, the EDS market was going to be reduced to one of selling } commodity EDS components. Rather than passively await this fate, we } decided to be bold and enter the SEM market ourselves, since that is } where we thought the real long-term action and opportunity lay. Though } we were very anxious about the impact on our EDS sales, we felt that we } were at a "window of opportunity" where we could pull this off, but that } the window was sure to close. With this in mind, we decided to make } our move. The final go-ahead was made with the full understanding of } 'Van' Vandenheuvel (general manager of TN), Bill Buffo (president of TN } and Tracor VP of the Instruments Group) and Frank McBee (president of } Tracor) that this was going to have a negative impact on EDS sales, but } it was felt that we could weather this as Tracor was a very healthy } company at that point. Since TN had been a consistently profitable } company for the past 20 years, it was Tracor's position that it was } appropriate to invest short-term profits for a future position in a } market where we thought we had something unique to offer. } } Central to our strategy was a conviction that we could provide a } substantially better value by designing a fully integrated SEM and thus } offer both better price and productivity than was possible by the } "piecemeal" approach of installing an EDS/automation system on a } stand-alone SEM manufactured by someone else. I think we actually } delivered on this rather well. Despite the fact that ADEM was indeed a } rather complicated system, to be fair you need to compare it with the } "glued together" systems it was designed to replace. Compared to these, } ADEM was a bargain, simple to run and maintain, and highly productive. I } am told that the instruments in the field have proven to be reliable, } and had the development and support team remained intact, I suspect the } instrument could have enjoyed a long life. So what went wrong? Having } thought about it a lot in the subsequent years, I note three key } "surprises" which ultimately played a large role in dooming our efforts: } } (1) Introduction of field-emission SEM technology; } (2) The leveraged buyout of Tracor; and } (3) The outbreak of "global peace". } } Field emission was a technology breakthrough we didn't anticipate. We } were able to keep ADEM under wraps until its market introduction in the } summer of 1987, and thus stave off erosion of EDS sales until we were } ready to make our move. The introduction was quite successful, and } those present at the Baltimore EMSA/MAS introduction may remember the } SRO crowds it drew. There was no question that ADEM made a big splash. } However, it was around this time that Hitachi introduced their } field-emission SEM and this quickly became the technology where people } with ADEM-sized budgets thought they should spend their SEM dollars -- } particularly so in the semiconductor industry, which by now had evolved } into being the principal market for high-end SEMs. We had targeted the } existing thermionic SEMs of the early 80's, and felt that we competed } quite well with the instruments of that type -- but field emission was a } "paradigm shift" for which we were unprepared. However, we were playing } for the "long term" and our development team knew we would have to work } both hard and smart to pull off what we were attempting. We started } work on our own field emission instrument. } } About this time, the financial bottom was also dropping out of Tracor. } With incredible ill-timing, a private investment group had finalized a } leveraged buyout of Tracor just days before the major market crash of } the late '80s. Then, the collapse of the Soviet Union depressed the } defense industry on which Tracor relied for much of their revenue. Also } around this time, McBee, Buffo, and Van all resigned, and with them went } the commitment to the product. Instead of struggling through a "trough" } as our SEM sales replaced lost EDS sales, as we expected, we found the } whole company fighting for survival. Eventually, Tracor was forced to } sell off the instruments group and the new management decided that they } had to jettison ADEM in late 1990 -- that's when I joined Rich Lee to } participate in his vision of a low-cost automatable SEM. } } Could ADEM have succeeded had these "surprises" not taken place? I like } to think so, but I also now see the huge obstacles we faced with a } clarity that only hindsight can provide. I think there was a certain } amount of "hubris" involved in our failure, and I accept the } responsibility for that. There were also some engineering decisions I/we } made which are questionable in retrospect. ADEM was loaded with } technological innovations, but I do wish in retrospect that I had been } more selective and focused -- had we kept the product simpler (and less } expensive) I think we might have vastly improved our chances. (That's a } hard-learned lesson that I am happy to say that we have been able to } apply to the development of the PERSONAL SEM at RJ Lee Instruments -- } and with a lot more satisfactory business outcome, I might add). And, } of course, had we had access to the kind of powerful/inexpensive PC } technology which is available today, we could have saved a whole lot of } engineering and provided a lot more bang for the buck -- another lesson } learned. } } The whole ADEM experience did leave me with deeply conflicted feelings. } On the one hand, I remain very proud of what our ADEM team accomplished, } and that we had the nerve to take on what we knew was a huge challenge. } But at the same time I am also painfully aware of the personal and } economic costs and the "bottom line" fact that we ultimately failed to } achieve our vision. The one thing I can say with certainty is that the } ADEM team was spectacularly competent and, based on their dedication and } effort, they deserved an outcome far better than what I was able to } deliver. } } A final bit of irony -- while we were laboring in our secret quarters in } an industrial park miles from the main TN plant, the ADEM team } occasionally noted the weird activities of a nearby startup. It turned } out they were marketing collector dolls. Fast-forward seven years. I } have an article on my desk from our local Pittsburgh paper describing } the phenomenon that the Pleasant Company "American Girl" dolls have } become -- stating that the company turned a PROFIT of $80 million last } year! Mattel recently purchased the company for something north of $300 } million, as I recall. ADEM has been a defunct product for years. Is } there a lesson here? } } Fred Schamber } RJ Lee Instruments Limited } ....................... } mailto:fhscham-at-SGI.NET --------------C13D3920A222C3F36012EB4A Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Timothy Moeller Content-Disposition: attachment; filename="vcard.vcf"
Hello to those in listserv land - I'm a long time lurker (two or more years) first time poster. There have been bunches of good stuff on the list that have saved me time, money and effort, for which, thanks to all. Now, I actually have some questions that I have not seen addressed as yet. A bit of background on me: I manage Arizona State University's Life Science EM Lab and have for over twenty years. We are a teaching and research lab with a STEM/EDS, a TEM, and an old, non-functional SEM which is the crux of all my questions. We are writing a grant proposal for a replacement for the SEM and have been very interested in a Philips/FEI ESEM FEG for its apparent resolution, flexibility, and all round usefulness. The questions we have are: 1) Can work be done on unfixed, hydrated biological specimens without cell walls such as animal tissue and/or cultured cells? 2) Can this type of work be done at high resolution, either at high or low KeV and how much beam damage does one expect? 3) Is manipulation of water vapor pressure / temperature (w/Peltier stage) sufficiently precise to do this type of work routinely? 4) Do unprotected cells simply explode when introduced into the scope? 5) Does anyone out there have such a scope in a biologically based lab or know someone who does?? 6) We have searched the literature and found virtually NO bio based papers in our meanderings. Are there papers we are missing? (we do know that FEG ESEM is new and not many are about - how about the older Electroscan W filament or LaB6 types? Are they amenable to high-ish resolution work on bio stuff? OK. TOO much bandwidth for a first post. Thanks in advance for any and all info or leads. --------------------------------------------------------------- William P. Sharp Department of Botany Arizona State University Tempe, AZ 85287-1601 --------------------------------------------------------------- voice (602) 965-3210 | fax (602)965-6899 | email wsharp-at-asu.edu
Sorry, Mr.Passero, This community doesn't need to see an attached photo of a microscope. Please use your computer for better purposes.........
Joseph Passero wrote:
} Leitz, Model SM-LUX, binocular head with 10x Periplan eyepieces. The } 170mm tube length objectives are: 3.5/0.10, 10/0.25, 45/0.65, and } 100/1.30 oil, the 45x and 100x are spring loaded noses. All the } objectives lens are Leitz. The microscope has a light source with } transformer. Everything works smoothly and the optics are crisp and } clear. There is a small wear spot on the frame where the paint has been } rubbed away by the support inside the case. Otherwise it is in very good } condition. } } See attached photo file. } } Entertaining Offers } } Thank You } } Joseph Passero } jp-at-spacelab.net } } ------------------------------------------------------------------------ } [Image]
This posting is only relevant to those who from time-to-time import chemicals from overseas. Chuck Garber's posting is rather similar to one he produced here about a year ago. Dangerous Goods shipping rules are no secret to any EM supplier. The "shipping DG's in limited quantity" rules are of little use, in fact BDMA is about the only chemical were one could save the DG fee, but it still must be airfreighted. Note A$1 = US$0.58
Here is a part of our dangerous goods notes from our online - Dangerous goods in limited quantities: Most chemicals can be shipped overseas in limited quantities. This rule allows up to 500ml or grams but packed in 30ml/g lots. Only two of our DG chemicals, osmium tetroxide and propylene oxide, cannot be shipped under that clause; for shipping, these two chemicals are always treated as dangerous goods. All our other DG chemicals could be airfreighted to overseas destinations, but never posted. However, it is not viable to ship 500g quantities dispensed in 30g quantities. Hence this rule is only useful for a few items, for instance BDMA, which is normally shipped in 100ml quantities. This chemical is also available as 4x25ml for overseas customers. When shipping DGs in limited quantities, the saving is only the dangerous goods fee, which is currently A$60. We would still need to charge for airfreight (whereas we Post (orders } $50) for free.
Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Tuesday, 18 August 1998 0:17, Garber, Charles A. [SMTP:cgarber-at-2spi.com] wrote: ..... ...... On the other hand, and it is not widely known, by taking } advantage of } certain small quantity shipping exemptions, one can still } get their BDMA in } such markets relatively inexpensively, but by using a } different packaging } approach, and this approach is explained on our website } under "Hazardous } Materials: Good News and Bad News". For the US domestic } market, these } differences are very small and are not significant. The } bigger differences } come into play when making shipments over international } boundaries. } } Chuck
William Chris Gilpin {cgilpin-at-fs1.sem.man.ac.uk} is probably the most experienced biological ESEM user and I know he sometimes posts to this list so I expect you will hear from him. I hope he provides us all with a sample of references to bio ESEM. I suggest you contact Trisha Rice at Electroscan {trice-at-electroscan.com} as she is probably the most experienced at ESEM FEG and will be able to answer your FEG specific questions. Unfixed hydrated biologicals can be examined with enough control to keep the specimen hydrated. We have a LaB6 ESEM and do mainly materials and particles with resolution comparable to conventional SEM, I suspect the same is true for biologicals. Good luck, Scott
Not an endorsement, no financial stake, just a satisfied customer.
} interested in a Philips/FEI ESEM FEG for its apparent resolution, } flexibility, and all round usefulness. The questions we have are: } 1) Can work be done on unfixed, hydrated biological specimens without cell } walls such as animal tissue and/or cultured cells? } 2) Can this type of work be done at high resolution, either at high or low } KeV and how much beam damage does one expect? } 3) Is manipulation of water vapor pressure / temperature (w/Peltier stage) } sufficiently precise to do this type of work routinely? } 4) Do unprotected cells simply explode when introduced into the scope? } 5) Does anyone out there have such a scope in a biologically based lab or } know someone who does?? } 6) We have searched the literature and found virtually NO bio based papers } in our meanderings. Are there papers we are missing? (we do know that FEG } ESEM is new and not many are about - how about the older Electroscan W } filament or LaB6 types? Are they amenable to high-ish resolution work on } bio stuff?
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
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A Postdoctoral position in electron microscopy is available at Unilever Research Port Sunlight Laboratory Wirral UK.
Qualifications: Recent PhD in Material Science or related area. High level of hands-on experience in TEM and proficient ultramicrotomy expertise are essential for success in this role. Cryo experience at this level is desirable. Excellent communication, interpersonal and team working skills required.
The position is for one year in the first instance.
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I too would appreciate information on this subject. Please post responses to the listserve or, Bill, would you mind summarizing responses and then posting the summary?
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------
Hello to those in listserv land - I'm a long time lurker (two or more years) first time poster. There have been bunches of good stuff on the list that have saved me time, money and effort, for which, thanks to all. Now, I actually have some questions that I have not seen addressed as yet. A bit of background on me: I manage Arizona State University's Life Science EM Lab and have for over twenty years. We are a teaching and research lab with a STEM/EDS, a TEM, and an old, non-functional SEM which is the crux of all my questions. We are writing a grant proposal for a replacement for the SEM and have been very interested in a Philips/FEI ESEM FEG for its apparent resolution, flexibility, and all round usefulness. The questions we have are: 1) Can work be done on unfixed, hydrated biological specimens without cell walls such as animal tissue and/or cultured cells? 2) Can this type of work be done at high resolution, either at high or low KeV and how much beam damage does one expect? 3) Is manipulation of water vapor pressure / temperature (w/Peltier stage) sufficiently precise to do this type of work routinely? 4) Do unprotected cells simply explode when introduced into the scope? 5) Does anyone out there have such a scope in a biologically based lab or know someone who does?? 6) We have searched the literature and found virtually NO bio based papers in our meanderings. Are there papers we are missing? (we do know that FEG ESEM is new and not many are about - how about the older Electroscan W filament or LaB6 types? Are they amenable to high-ish resolution work on bio stuff? OK. TOO much bandwidth for a first post. Thanks in advance for any and all info or leads. --------------------------------------------------------------- William P. Sharp Department of Botany Arizona State University Tempe, AZ 85287-1601 --------------------------------------------------------------- voice (602) 965-3210 | fax (602)965-6899 | email wsharp-at-asu.edu
RFC822 header -----------------------------------
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Dear Don,
To the contrary: a photo of a microscope often helps in identifying it and therefore in making an educated decision.
I would like to annonce and post the following new open position in the Analytical Instrumentation Facility at NCSU. Please note that this position is in addition to the position recently posted, which I have copied below for clarity. With the two microscopy positions open, we hope to find complenentary individuals to complement the expertise and skills of our existing staff. More details of our lab facilities and program can be at http://spm.aif.ncsu.edu/aif/index.html
Research Assistant / Microscopist
Announcing an opening as of Oct. 1, 1998 for the postion of Research Assistant at the North Carolina State University Analytical Instrumentation Facility (AIF).
Qualifications must include an BS as the minimum degree with higher degree desired or equivalent experience in a materials related discipline (non biological) along with hands on experience with optical metallographs, XRF, XRD, SEM, SEM/EDS, TEM and sample preparation instruments such as sample coaters, ion millers, mounting presses and grinding, polishing and sectioning instruments. Required skills include: demonstrated analytical problem solving capabilities in a multiuser analytical facility environment, extensive hands on experience with the above techniques along with their tools and accessories (e.g. metallographic, TEM, XRF, XRD and other specimen preparation, analysis of metallographic and other samples, etc.); teaching/user training; and familiarity with modern instrumentation, computer systems and experience with vacuum systems. Responsibilities include: operation and maintenance of optical metallographs, ion millers, X-Ray diffractometers and sample preparation devices such as mounting presses and grinding, polishing and sectioning devices, etc; scheduling of access to and oversight of the above instrumentation; and user training and assistance. Other responsibilities include operation of SEM and TEM and assistance with the teaching of electron microscopy laboratory classes and assistance with other graduate level engineering classes. Qualifications must include an BS as the minimum degree with higher degree desired or equivalent experience in a materials related discipline (non biological) along with hands on analytical experience in a multiuser analytical facility environment,
Please send resume and names of references to: Phil Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 118A EGRC; 1020 Main Campus Drive; Raleigh, NC 27695-7531
North Carolina State University is an Equal Opportunity, Affirmative Action Educator and Employer. Minority and Female Applicants are especially encouraged.
Previously posted Position Open -- (still avaliable)
Microscopy and Microanalysis Research Assistant
An immediate position is open for an SEM and TEM microscopist at the North Carolina State University Analytical Instrumentation Facility (AIF) as a retirement replacement.
Duties and responsibilities include: Teaching of laboratories; user training and assistance; and instrumentation development, modification and maintenance; and analysis of a wide variety of samples. An MS or equivalent experience in a materials related discipline along with three or more years hands on experience with SEM and/or TEM is required. Required skills include: extensive hands on experience with SEM, TEM and related techniques and accessories (e.g. specimen preparation and associated tools such as evaporators, etc.); teaching/user training; and familiarity with modern electronics, computer systems and experience with vacuum systems.
Please send resume and three letters of reference to: Phil Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 118A EGRC; 1020 Main Campus Drive; Raleigh, NC 27695-7531 or email PRUSSELL-at-NCSU.EDU.
North Carolina State University is an Equal Opportunity, Affirmative Action Educator and Employer. Minority and Female Applicants are especially encouraged.
-- Phillip E. Russell Professor, Materials Science and Engineering Director, Analytical Instrumentation Facility Box 7531, Room 318 EGRC 1010 Main Campus Drive North Carolina State University Raleigh, NC 27695-7531
Ronnie Houston wrote: } } We are looking at proliferating cells in animal tissues and, at present, are giving the animals (goats) 100mg/Kg body weight IV two hours prior to sacrifice.
This dose may be cytotoxic, it is in mice.
} One of our researchers has suggested giving the animals } another dose approx 5-7 days prior to this one.
How will that help?
} Before we consider this, we wish to know a couple of things about BrdU, } and have been unable to obtain any information from our supplier } (Sigma). How long does BrdU remain in the bloodstream before being } excreted,
BrDU is no longer available within one hour, probably unavailable within 30 minutes in mice. That can be a good thing depending on your experimental design.
} and which organ(s) is responsible for BrdU breakdown and } excretion.
My guess would be liver and/or kidney but it might be broken down by the lungs as well. Might vary with the species.
} What is the half-life of BrdU, ie, how long can the BrdU be } detected immunocytochemically in cells after initial dosage?
I don't think half-life is the correct term but cells hold on to BrDU for a long time. It will be diluted below the point of IHC detection after 2 or maybe 3 cell divisions.
} What accumulative effect will the BrdU have at this dose if given several times over a short time frame, say three weeks?
Might slow the cell cycle in targeted cells or even kill them.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
I need to do some SEM imaging of uncoated/unexposed photoresist at low kV. Does anyone have any recipies or advice on this? This is on a PMMA coated Si wafer. Thanks Bill Neill
Does anyone know where I can find a positioner for microscope specimens with an overall travel range of a few microns and step size in the nanometer range?
****************************** Jim Haley e-mail: haley-at-i-cubeinc.com ******************************
William and the list, My thanks firstly to Scott for the recommend. I have been using a tungsten ESEM for biological work for the last 6 years. Although I have published little in print I have presented at numerous meetings. So only abstracts in the literature I'm afraid. OK a couple of papers on EDX in ESEM and some on pharmaceutical applications but not strictly biological. Now to your questions!
1) Yes you can work on unfixed hydrated samples. 2) depends on what you call high resolution 3) manipulation of hydration is easy with a little practice. 4) no cells do not explode. 5) Yes, I have a tungsten E3 and have used a.n.other FEG ESEM for biology. 6) See my opening my remarks
Those are the simple answers! To adequately cover the detailed answers would take up much space. Here are my main thoughts for other list readers. I can give a fuller account off list if it will help.
Cells often look very different when examined by ESEM compared to Hi VAC. I have always taken this to represent a "truer" view of biological surfaces - very smooth with little fine topography. I assume that extracellular matrix is well preserved with ESEM giving this appearance. Hence the comment on resolution. The microscope is capable of matching Hi Vac for resolution but many biological structures do not have that much detail to show. Aesthetically disappointing but with correct interpretation valuable information on structure. Low KV is difficult with a tungsten ESEM but not impossible. I routinely image at 5KV. Modification of the detector (as per Brendon Griffin)allows imaging at a few hundred volts. Beam scattering in the chamber gas is the problem and simple physics shows that the more electrons you start with (FEG } LaB6 } W)) the more will reach the sample. You are right to highlight hydration as being important. Very fine control is easy with the older instruments but I worry about the vacuum increment steps in the windows driven new instruments. 0.1 Torr or 10 Pa is not really fine enough but still possible. I have come to the conclusion that many cells are best viewed hydrated but after a short fixation in glutaraldehyde and rinse in water. This has a number of benefits. Fresh samples which arrive at 5 o'clock on a Friday can be examined on Monday morning! Beam damage is greatly reduced. Finally cells cannot be examined from culture medium or buffers because the salts precipitate out following excess liquid removal obscuring anything of interest. Unfixed samples can be examined but my experience has shown that the structure is pretty similar in both cases. It is dehydration that causes the problems not fixation. Remember that cryo stages are also available for ESEM - just use different chamber gas (see papers from the Cavendish group in Cambridge). Cells tolerate the vacuum (at least 4.6 Torr to be hydrated above freezing) well. Electroscan/FEI/Philips have video on moving insects in their promotional material.
One final point to make. If you have an ESEM you also have a very capable Hi Vac and controlled pressure instrument as well. Don't fall into the trap of should I get ESEM OR conventional - you can have both in one instrument! Another final point! The images obtained in ESEM mode should be taken as another piece of information in a bigger picture and should be seen as complementary to other SEM techniques.
My own opinions as a satisfied customer (how about a free upgrade Ralph?)
Chris Gilpin Experimental Officer Biological Sciences EM Unit University of Manchester Oxford Road Manchester M13 9PT Phone +44 (0)161 275 5170 Fax +44 (0)161 275 5171
---------------------------------------------------------------------------- ----------------- William P. Sharp wrote
We are writing a grant proposal for a replacement for the SEM and have been very interested in a Philips/FEI ESEM FEG for its apparent resolution, flexibility, and all round usefulness. The questions we have are: 1) Can work be done on unfixed, hydrated biological specimens without cell walls such as animal tissue and/or cultured cells? 2) Can this type of work be done at high resolution, either at high or low KeV and how much beam damage does one expect? 3) Is manipulation of water vapor pressure / temperature (w/Peltier stage) sufficiently precise to do this type of work routinely? 4) Do unprotected cells simply explode when introduced into the scope? 5) Does anyone out there have such a scope in a biologically based lab or know someone who does?? 6) We have searched the literature and found virtually NO bio based papers in our meanderings. Are there papers we are missing? (we do know that FEG ESEM is new and not many are about - how about the older Electroscan W filament or LaB6 types? Are they amenable to high-ish resolution work on bio stuff? OK. TOO much bandwidth for a first post. Thanks in advance for any and all info or leads. ---------------------------------------------------------------
Sodium borohydride, glycine, ammonium chloride are all used for pretreatment of sections for LM and TEM immunocytochemistry. Does anyone know the difference in actions between these? Are there any preferences for better ultrasctructural preservation among the three? I cannot seem to find this answer anywhere. Would anyone know? Or know where to find the answer?
I agree with Barbara. It would appear that one of the major on-line laboratory equipment reseller services also agrees - see www.labx.com - where most equipment advertisements are accompanied by a picture. The latter aspect is especially important at the labx site in the selling of microscopes as there are often several of the same scope, or more importantly several variants of a model for sale at the same time. However, I do agree that we do not need, and should not be subjected to unsolicited (file) attachments to e-mail.
Winton Cornell
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
Are there any Soil Scientists out there that can help my boss determine = the best way to pH the soil from his recently purchased farm. We were = trying to determine if adding tap water, dionized water, or a standard pH = buffered solution to the dry soil could influence an accurate pH. I know = this isn't rocket science, but I am sure there is a proper way to do this = and a lot of ways to get bad data. Thanks Linda Fox lfox1-at-wpo.it.luc.edu
Leica CLSM - confocal microscope (inverted). The system includes: a Leitz Fluovert inverted microscope, objective lenses, filters, krypton/argon laser, Motorola computer (running OS9) with Leica ScanWare software, and 3 monitors. Also for sale is a Focus Graphics image recorder. All reasonable offers will be considered.
If interested, please contact:
Scott Henderson, Ph.D. Director of Microscopy, Dept. Cell Biology & Anatomy, Mount Sinai School of Medicine, Box 1007, 1 Gustave L. Levy Pl., New York, NY 10029-6574
Scott Henderson, Ph.D. Director of Microscopy, Department of Cell Biology & Anatomy, Mount Sinai School of Medicine, Box 1007, One Gustave L. Levy Place, New York, NY 10029-6574
We have had pretty decent luck using an accelerating voltage around 600 - 800 eV. That is about the extent of our recipe.
John Staman Analytical Lab LSI Logic formerly Symbios Inc.
} -----Original Message----- } From: Bill Neill [SMTP:billneill-at-csi.com] } Sent: Tuesday, August 18, 1998 4:31 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Imaging of Photoresist } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I need to do some SEM imaging of uncoated/unexposed photoresist at low kV. } Does anyone have any recipies or advice on this? This is on a PMMA coated } Si } wafer. } Thanks } Bill Neill } }
Dear Bill, John Cole of Hitachi Canada gave me a nice sheet of: "SEM Specimen Charging Threshold Energies of Polymers and Semiconductor Materials", which lists the upper limit for PMMA resist as 1.60 kV. This means you should not get charging of this material if you stay below this accelerating voltage. You can contact him for the complete list at: http://nsctoronto.com/ or: e-Mail nscsales-at-nsctoronto.com . You wrote: } } I need to do some SEM imaging of uncoated/unexposed photoresist at low kV. } Does anyone have any recipies or advice on this? This is on a PMMA coated Si } wafer. } Thanks } Bill Neill } Best of luck, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
I suppose the addition of the photo does slow down the message time, but I was glad to see it. I did'nt get into this thread until after the first message so I do not know how long the message took to come down, but since I had an interest in seeing the unit I had the person send me a copy of it to my mailbox. As they say, A paicture is worth a thousand words and although I probably remember more models of microscopes by their given names my memory is not perfect. FOr some reason I though te sm lux was a white or grey colored newer microscope so by getting the photo it let me know what I was looking at. Perhaps in the frame of not having slow messages it is best to request the photo from the sender? If it was a small photo the transfer time wopuld not be long. Perhaps with the initial message just a small thrubnail and then request from the sender a large fit to print and frame over our desk version on request.
I am rambling so I will quit and go drink half a pot of coffee.
I will like to know the full use and range for testing materials using a = scanning electron microscope model no. JSM-35C,at present = microstructure, particle morphology and micrographs are being carried = out.We will like to know the other tests we can carry out.=20
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.2016.0"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I will like to know the full use and = range for=20 testing materials using a scanning electron microscope model no. = JSM-35C,at=20 present microstructure, particle morphology and micrographs are being = carried=20 out.We will like to know the other tests we can carry=20 out. {/FONT} {/DIV} {/BODY} {/HTML}
} Date: Tue, 18 Aug 1998 14:00:38 -0600 (MDT) } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } To: postmessage {Microscopy-at-sparc5.microscopy.com} } Subject: Naborohydride, Glycine, A.Chl???? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi, } } Sodium borohydride, glycine, ammonium chloride are all used for } pretreatment of sections for LM and TEM immunocytochemistry. Does anyone } know the difference in actions between these? Are there any preferences } for better ultrasctructural preservation among the three? I cannot seem } to find this answer anywhere. Would anyone know? Or know where to find } the answer? } } Thank you, } Hildy } {hcrowley-at-du.edu}
Hi Hildy, et al.
NaBH4 is used as an etching agent on epoxy sections to clear away some of the resin and expose antigens for immunolabeling. (Not sure how used in LM). Trouble is, it can eat away your antigen--and even your section if too strong or left too long. Worked OK for me at ~1% for ~ 10 min, but method requires presence of lots of antigen and strong antibody. I prefer acrylic resins which don't have to be etched, or even better ultrathin cryosections--which we routinely do now.
Glycine and NH4Cl are used to quench the aldehydes before immunolabeling--supposedly frees up antigens for more access by antibodies. I don't remember the glycine conc, but the NH4Cl is usually used between 50-100 mM for about 10 min (on sections--may differ for LM??).
Cheers, S
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I will like to know the full use and range in testing materials on an = scanning electron microscope model no. JSM-35C.At present tests that can = be carried out are Microstructure, Particle Morphology and Micrographs.=20
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.2016.0"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I will like to know the full use and = range in=20 testing materials on an scanning electron microscope model no. = JSM-35C.At=20 present tests that can be carried out are Microstructure, Particle = Morphology=20 and Micrographs. {/FONT} {/DIV} {/BODY} {/HTML}
In addition, sodium borohydride is also a strong reducing agent, and can be used on fixed tissue to reduce aldehyde groups, thereby diminishing autofluorescence. In fact, it is a stronger reducing agent than the other two, so it can be used when they are inadequate to the task. A very good article on its use is:
Eldred WD, Zucker C, Karten HJ, Yazulla S. 1983. Comparison of fixation and penetration enhancement techniques for use in ultrastructural immunocytochemistry. J Histochem Cytochem 31:285-292.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
} On Tue, 18 Aug 1998, HILDEGARD CROWLEY wrote: } } } Date: Tue, 18 Aug 1998 14:00:38 -0600 (MDT) } } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } } To: postmessage {Microscopy-at-sparc5.microscopy.com} } } Subject: Naborohydride, Glycine, A.Chl???? } } } } Hi, } } } } Sodium borohydride, glycine, ammonium chloride are all used for } } pretreatment of sections for LM and TEM immunocytochemistry. Does anyone } } know the difference in actions between these? Are there any preferences } } for better ultrasctructural preservation among the three? I cannot seem } } to find this answer anywhere. Would anyone know? Or know where to find } } the answer? } } } } Thank you, } } Hildy } } {hcrowley-at-du.edu} } } Hi Hildy, et al. } } NaBH4 is used as an etching agent on epoxy sections to clear away some of } the resin and expose antigens for immunolabeling. (Not sure how used in } LM). Trouble is, it can eat away your antigen--and even your section if } too strong or left too long. Worked OK for me at ~1% for ~ 10 min, but } method requires presence of lots of antigen and strong antibody. I } prefer acrylic resins which don't have to be etched, or even better } ultrathin cryosections--which we routinely do now. } } Glycine and NH4Cl are used to quench the aldehydes before } immunolabeling--supposedly frees up antigens for more access by } antibodies. I don't remember the glycine conc, but the NH4Cl is usually } used between 50-100 mM for about 10 min (on sections--may differ for } LM??). } } Cheers, } S } } Sara E. Miller, Ph. D. } P. O. Box 3020 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-8735
We are attempting to fix chick forebrain neurons for TEM. Several cells have been processed and embedded in Marglass. A graded series was not used to dehydrate or embed, the blocs had serious infiltration problems. To solve this I suggested using a graded series (with EtOH) and Spurtol. They tried and the Spurtol etched the plates. The plates are poly-L-lysine coated tissue culture dishes made by Falcon (Becton Dickinson). Since the Spurtol was a problem, they have been experimenting with Marglass and a graded EtOH series. However, when they try this, the plates are again etched. Any suggestions?? Change resin? Change plates? suggestions on infiltration timing would also be helpful
Thanks... Sally
Sally Burns Center for Electron Optics B5 Pesticide Research Center (517) 355-5004
I am about to do some LM autoradiographic studies and would like to know if anyone has any experience with chemography effects when sections are pre-stained with Safranin O. In the past I have successfully used Hematoxylin and PAS but prefer Hematoxylin + Saf O these days because I get a more intense staining. I am aware, however, that some dyes (e.g., Tol Blue) can't be used in pre-staining sections for autoradiography due to chemography. To save time and emulsion, I would appreciate hearing from anyone who has either successfully or unsuccessfully used Saf O prior to autoradiography. TIA Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Postdoctoral Position in Environmental Catalysis/HREM
A postdoctoral position is available at Northwestern University to work on Environmental Catalysis. The work will involve in-situ work using a unique UHV-HREM and gas treatment of various samples (see http://www.numis.nwu.edu for some information about the hardware). A strong background in HREM and crystallography is highly desirable, and experience in catalysis and/or surface science will be significant; extensive experience in other types of TEM are an alternative. To apply, send an email to L. D. Marks at ldm3-at-apollo.numis.nwu.edu including a CV and the names of referees.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 email: ldm3-at-apollo.numis.nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Ron Anderson has asked that I request a copy of of the complete list (of upper limits for PMMA resist). Please mail it to the following:
Ronald M. Anderson Treasurer, Microscopy Society of America c/o I.B.M. Corporation Analytical Services Group 1580 State Route 52, Mail Stop E40 Hopewell Jct., NY 12533-6531
Thank you so much .
Regards,
Colleen Marie Genadry Mail Stop E40, B/630, Dept. 13WA Lotus Notes ID: IBMUSM08(CGENADRY) VM ID: FSHVM1(CGENADRY) T/L532-2664 } From Outside call (914)-892-2664 FAX #: (914)-892-2003
I'm looking for a company that recoats phosphorus TEM screens. In particular, a company that uses a greener phosphorus than more yellow. In the past I used a company called Grant Scientific but have been unable to locate them.
Does anyone know how to get commercially available actin to disperse as F-actin? We have not had good luck getting a lyophilyzed powder of porcine actin (Sigma) into solution as dispersed F-actin filaments. We usually get quite a bit of clumping with only a small amount of the actin appearing as single filaments. We have tried sonicating, gentle swirling for several hours, vortexing, and ammonium hydroxide.
As we understand it, the lyophilyzed product as purchased is G-actin that will polymerize to F-actin once in solution. However, we do not believe that the G-actin is going into solution, or at least that only a very small proportion of it is.
Any help would be greatly appreciated.
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
Are there companies which deal with buying and selling of 2nd hand TEMs? If so, I would appreciate contact names/emails/telephone numbers.
Thanks.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Has anybody had any experience of using the Nikon F90X with a Zeiss Axioskop? I'm not keen on the Contax 167MT that Zeiss want to sell me so I'm looking for an alternative. My main application is B/W photography of fluorescence staining.
If people are worried about file attachments, they shouldn't worry about it. Simply do NOT extract the file and if your mailer does it automatically, one can switch that option off somewhere in the options...usually just a flag switch. Usually I never extract attachments by default, because most of the time they are a waste of time and meant to be funny or something, and indeed it may slow your system down. Off the topic really, but just my two cents, which in our currency is worth sweet blow all, to calm down some some frantics on this list.
Democratically,
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
A simple answer to your question is a bit beyond the capability of this list server. However, we have several consultants on staff who can help you with your specific application. Please either visit our website ( { {http://www.MME-Microscopy.com/education} ) or call our office.
Best regards,
At 02:34 PM 8/19/98 -0300, MATERIALS-CARIRI wrote:
} } } }
{excerpt} {smaller} I will like to know the full use and range in testing materials on an scanning electron microscope model no. JSM-35C.At present tests that can be carried out are Microstructure, Particle Morphology and Micrographs.
Sally Burns wrote: } } We are attempting to fix chick forebrain neurons for TEM. Several cells have been processed and embedded in Marglass. A graded series was not used to dehydrate or embed, the blocs had serious infiltration problems. } To solve this I suggested using a graded series (with EtOH) and Spurtol. They tried and the Spurtol etched the plates. The plates are poly-L-lysine coated tissue culture dishes made by Falcon (Becton Dickinson). } Since the Spurtol was a problem, they have been experimenting with } Marglass and a graded EtOH series. However, when they try this, the plates are again etched. } Any suggestions?? Change resin? Change plates? suggestions on infiltration timing would also be helpful } } Thanks... Sally } Sally Burns } Center for Electron Optics } B5 Pesticide Research Center } (517) 355-5004 } burnssal-at-pilot.msu.edu
Sally:
I embed cultured neurons or glia in an Epon substitute without any problems. While I use EMbed 812 from Electron Microscopy Sciences I'm sure an Epon substitute from another vendor would work. Perform fixation, osmication, rinse, dehydrate as usual. When you finish with 100% alcohol (or even 95%) then go to 100%:resin 2:1, then 1:2, then two changes of pure resin. I usually get compulsive and leave the tissue culture dish in a vacuum dessicator overnight to get all of the solvent out of the last change. Then polymerize as usual. NO propylene oxide! "Epon" is completely miscible with ethanol even with a bit of water left over, it just does not mix as quickly as with prop. oxide. "Epon" (and its components) reacts very slowly with tissue culture plastic, Spurr and its components react very quickly (I actually did the experiments!) E-mail me if you need more advice.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
For my recent project I have to section perfused and cryoprotected rat brains 40 microns and do immunohistochemistry on floating sections. Sometimes the sections have to wait a couple of weeks before I do the immuno on them. I usually keep them in PBS in the fridge but always worry that it may lose some of it's antigenicity. In what would you recommend the sections be stored? Thank you, Lilith ------------------------------------------------------- Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
RESEARCH ELECTRON MICROSCOPIST to manage institutional core EM Facility and carry out all duties related to daily operation of the facility, including maintaining and using electron microscopes, preparing specimens, data collection, advising researchers on EM approaches, ordering supplies, record keeping and billing, etc. Requires minimum of Bachelor's degree and several years experience in biological electron microscopy; up to Ph.D. level considered. Expertise in several of the following techniques required: thin sectioning, immunolabelling, cryo-EM, negative staining, metal shadowing, cryosectioning, freeze fracture/etch, rapid freezing and freeze-substitution, EM autoradiography, in situ hybridization. Salary is highly competitive and commensurate with experience. Send CV and names of three references to Dr. George Witman or Dr. Roger Craig, Department of Cell Biology, University of Massachusetts Medical School, 55 Lake Ave. North, Worcester, MA 01655. Email: Roger.Craig-at-ummed.edu or George.Witman-at-ummed.edu. The University of Massachusetts Medical School is an equal opportunity employer.
Hi Bob, My experience with lyophilization is that unless precautions are taken with the protein (glycerinization or ammonium persulfate precipitation) that you will end up with a bunch of agglomerated, insoluble blobs that will never go into solution. You might try adding some NaCl (1% w/v) or ammonium persulfate (same) to see if this will enhance suspension of the molecules. Did you call the Sigma tech reps? With some enzymes (glucosyl transferases from bacteria), we have had good luck storing the enzyme solution under a layer of toluene. Lasts for months. Let us know what you find out.
} Does anyone know how to get commercially available actin to disperse as } F-actin? We have not had good luck getting a lyophilyzed powder of porcine } actin (Sigma) into solution as dispersed F-actin filaments. We usually get } quite a bit of clumping with only a small amount of the actin appearing as } single filaments. We have tried sonicating, gentle swirling for several } hours, vortexing, and ammonium hydroxide. } } As we understand it, the lyophilyzed product as purchased is G-actin that } will polymerize to F-actin once in solution. However, we do not believe } that the G-actin is going into solution, or at least that only a very small } proportion of it is.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
---------------------- Forwarded by Sylvie Verdon/CRYOLIFE on 08/20/98 01:34 PM ---------------------------
Sylvie Verdon 08/20/98 01:21 PM
To: Microscopy-at-MSA.Microscopy.Com cc:
We do see some dirt here at the Ag Research Center, so I asked someone who had made soil pH measurements. Her reply:
It's a 1:1 dilution.
We check the pH of the field soils by using 10 g soil:10 ml 0.01M CaCl2 in a 15-ml screw cap test tube. Shake well, spin & measure pH. You can use d. h20 as we used to before changing to the CaCl2.
lorraine
via Leonard R. Corwin Fort Dodge Animal Health Princeton, NJ 08543-0400
The following position is available in our Department. (Please note that all responses should be addressed to Human Resources at the address given in the announcement, and not to me.)
Dr Gillian M. Bond Department of Materials & Metallurgical Engineering New Mexico Tech
NEW MEXICO INSTITUTE OF MINING AND TECHNOLOGY seeks qualified applicants for the position of Departmental SEM/Electronics Technician with Materials and Metallurgical Engineering. Candidates should be capable of operating and maintaining scanning electron microscopes, (SEMs), as well as maintaining and repairing other items of equipment. The position will involve interaction with both faculty and students, and participation in current research activities. Experience in the operation and maintenance of SEMs and other electronic equipment is required. New Mexico Institute of Mining and Technology has a rural location in central New Mexico, with easy access to Albuquerque. The setting provides an excellent university environment, and the opportunity to work with very active research groups and high-caliber students. Interested persons should send a complete resume with details of prior experience. Submit application material to New Mexico Institute of Mining and Technology, Human Resources, 801 Leroy Pl, Wells Hall Box 92D Socorro, NM 87801. For information about New Mexico Tech, visit our web page http://www.nmt.edu/. E-mail applications NOT accepted. AAEOE
For the purposes of immunoEM, I would like to localize GFP (green fluorescent protein) at the EM level. I hope to find a source of anti-GFP antibody, preferably tagged already with 10 nm gold particles. I've seen commercial sources of polyclonal anti-GFP. Does anyone know of a gold-linked version? David H. Hall Center for C. elegans Anatomy Department of Neuroscience 1410 Pelham Parkway Albert Einstein College of Medicine Bronx, NY 10461
Lisa D. Brown University of Connecticut Physiology and Neurobiology Department Electron Microscopy Laboratory Box U-131, Rm 129 Beach Hall Storrs, Ct 06269-2131 Tel. (860)486-2914 Fax. (860)486-1936
Responding to the message of {85256666.00601C0D.00-at-cryont.cryolife.com} from "verdon.sylvie-at-cryolife.com"-at-Sparc5.Microscopy.Com:
} Sylvie Verdon } 08/19/98 02:37 PM } } To: Microscopy-at-MSA.Microscopy.Com } cc: } Subject: } } Has anyone done a trichrome stain with GMA? We are looking for a trichrome } staining procedure on GMA embedded Plastic sections for viewing on a light } microscope. } } Thanks
Sylvie,
How about just a general plant stain for GMA embedded tissue, like toluidine blue or methylene blue?
Or do you want to stain for something more specific??
Gib
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
I have used a variety of 35 mm cameras on several models of fluorescent microscope. My main concern with a camera such as the N90 is that unless the camera is parfocal to the eyepiece, the grain in its focusing screen makes composition and focusing difficult. A better solution would be a Nikon F3, F4, or F5. The normal viewing screen can be replaced with a clear "M" type screen and the prism replaced with a high magnification waist level finder. These cameras with appropriate screen will be easier to use than the N90. Also, these models of Nikon have mirror lockup switches, if you want to reduce the vibration induced by mirror slap. To install the Nikon in place of the Contax, I believe Zeiss sells suitable adaptor rings for their Axioscope.
Hope this helps. Brian Matsumoto Director of Microscopy Facility Neuroscience Research Institute University of California Santa Barbara, CA 93106
An understanding of the structure of foods and food products is essential to the efficient development of new processes or products. The future of the food industry is closely related to new developments in food microscopy.
This conference is an opportunity for those involved in food product development to hear the latest findings from key scientists in the field of food microscopy. The meeting will act as a forum to establish the needs of the food industry over the next decade, to discuss recent advances in food microstructural research and to evaluate new microscopical techniques.
This conference is aimed at food microscopists, food technologists and research scientists, particularly those involved in the development of new products or processes. Any company engaged in product development should be represented at the conference, to ensure that it is aware of the opportunities that advances in food microscopy can offer.
The aim of this conference is to:-
bring together food microscopists and food technologists to enable fundamental and applied research to be presented together;
highlight recent advances in food microstructural research;
provide a forum for discussion of microstructural studies relating to food;
identify the potential of new techniques.
Conference organisers: Mrs Kathy Groves and Dr Morag Saunders, Leatherhead Food RA; Dr Ashley Wilson, CCTR University of York.
There is still time for other papers to be submitted - please contact the conference organisers for further information.
KEYNOTE PAPER: Structure - the Only Thing that Matters? - Professor Peter Lillford, CBE, Unilever Research The challenge facing members of the food industry is to become architects of structure rather than processors of materials. This will need better application and novel approaches to microscopy.
Invited Paper: Cryogenic Environmental SEM of Ice Cream - an Introduction to the Principles of ESEM with Applications to Observations on Ice Cream - Brad Theil, Polymers and Colloids, Physics Department, University of= Cambridge
Food Microstructure using X-ray Projection Microscopy - John Judge and Peter Lillford
LUNCH
Chairman: David F. Lewis, SAC
The X-ray Microscope and its Applications to the Food Industry - Simon Burgess, Oxford Instruments
Probing Food Biopolymer Functionality with Atomic Force Microscopy (AFM) - Vic Morris, Institute of Food Research, Norwich (co-authors: A.P. Gunning, A.R. Kirby, A.R. Round, A. Mackie and P. Wilde)
The Environmental Scanning Electron Microscope - Speaker from Philips Electron Optics
Observations of Food Microstructure by Environmental Scanning Electron Microscopy - Debbie Stokes, Polymers and Colloids, Physics Department, University of Cambridge (co-author: A.M. Donald)
SESSION 2: APPLICATIONS OF MICROSCOPY TECHNIQUES IN FOOD RESEARCH
Chairman: Ashley Wilson, CCTR University of York
F Invited Paper: Applications of Variable Vacuum in Food Microscopy - Roger Angold, RHM Technology
F FTIR Microscopy in Troubleshooting for New Product Development - Hilary Holgate, RSSL
F Using Image Analysis and Confocal Microscopy Combined to Measure Deformation in Starch Matrices - Jeremy Addler, RHM Technology
F The Microscopy of Food. ESEM and Confocal Microscopy - Complimentary Techniques - Anthony Robinson, Polymers and Colloids, Physics Department, University of Cambridge
F Invited Paper: The Use of Electron Microscopy in the Investigation of BSE - Michael Stack, Veterinary Laboratories Agency
LUNCH and opportunity to view trade exhibition
SESSION 3: MICROSCOPY OF FOOD PRODUCTS, PROCESSES AND INGREDIENTS
Chairman: Roger Angold, RHM
Invited Paper: Microscopy - the Art of Food Technology - Professor Anne-Marie Hermansson, The Swedish Institute for Food and Biotechnology
Changes in Endosperm Structure during the Production of Popped Grain - Mary Parker, Institute of Food Research, Norwich
Confocal Laser Scanning Microscopy of Dairy Products and Ingredients - Methodology and Some Applications - Mark Auty, Dairy Products Research Centre, Moorepark, Ireland
Poster Exhibition and Opportunity to view Trade Exhibition
SESSION 3 (continued): MICROSCOPY OF FOOD PRODUCTS, PROCESSES AND INGREDIENTS
Chairman: Professor Anne-Marie Hermansson, SIK
Invited Paper: Brian Brooker, Institute of Food Research
The Use of SEM, Confocal Microscopy and Instrumental and Sensory Techniques in the Study of Biscuit Texture - Michael Edwards, Campden and Chorleywood Food RA
Crystallising Carbohydrates - Use of the Microscope in Understanding Crystallising Structures in Heat-resistant Chocolate and Powdered Sorbitol - Kathy Groves, Leatherhead Food RA
F Microstructural Changes of Food Products Following Inclusion of Non-starch Polysaccharides and Fibre Supplements: Implications to Dietary Nutrition - Karen Linton, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth (co:authors: C. Brennan and P. Moura de Magallraes)
LUNCH
Chairman: Brian Brooker, IFR, Reading; Mrs Kathy Groves, Leatherhead Food RA
Ultrastructural and Textural Changes in Heat-processed Fruits - Morag Saunders, Leatherhead Food RA
Cereal Structure: its Relationship to Raw Material Quality and End-product Utilisation - Charles Brennan, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth (co-authors: K. Linton, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth; D. Griggs, Pauls Malt Ltd; I. Cantrell, Pauls Malt Ltd; P.R. Shewry, IACR - Long Ashton)
Invited Paper: Perspectives on Food and Microscopy - David Lewis, SAC
CLOSE=09
ADMINISTRATION
Registration and closing times will be advised with the acknowledgement of your booking=09
Lilith- I would suggest freezing the tissue, probably prior to sectioning. Infiltrate tissue with OCT for 30-60 min. then Freeze, wrap in alum. foil and /or parafilm, and store in freezer (-20 C). I have done this in the past with human cornea, and antigenicity lasted months to years for the collagen Abs we worked with. -Mike
On Thu, 20 Aug 1998, Barry, Lilith wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear colleagues, } } For my recent project I have to section perfused and cryoprotected rat } brains 40 microns and do immunohistochemistry on floating sections. } Sometimes the sections have to wait a couple of weeks before I do the } immuno on them. I usually keep them in PBS in the fridge but always worry } that it may lose some of it's antigenicity. In what would you recommend the } sections be stored? } Thank you, } Lilith } ------------------------------------------------------- } Lilith Ohannessian-Barry } National Research Council } Institute of Biological Sciences } CANADA } Tel;613-993-6460 } Fax;613-941-4475 } e-mail; lilith.barry-at-nrc.ca } }
Fellow listmembers, Hope the day is going well for everyone. I am looking for some help. The department here (Dept. of Anatomy, National University of Ireland, Galway) is trying to set up a tissue culture facility and, although we have been lucky enough to get some grant money, we are having quite a bit of 'fun' trying to locate some equipment that we need at a price we can afford. Can anyone let me know of a good basic model of an inverted microscope that is relatively cheap and can be used for routine examination of cell cultures? We don't need any fluroescence or photographic capabilities, just the basic nuts and bolts microscope. Or, if anyone has a microscope that feels a bit of wanderlust and would like to live out its retirement in Ireland, we can offer a loving home!
I suppose emails directly to me would keep the list free from extraneous mails.
Since all the color correction, etc. has been done prior to the camera system, you should have no problem putting a Nikon camera on a Zeiss microscope. As a matter of fact, I think that Dr. Savile Bradbury (retired, Oxford), frequently used Nikon cameras on other types of scopes.
One place you may have to make a slight adjustment is on the parfocality (making sure that the microscope and the camera are both in focus simultaneously). One way to test is to set the microscope up for regular Koehler illumination (Brighfield) with the lowest mag objective (ex: 4x)using a thin, well-stained specimen. Mount the empty camera. (Zeiss has info on T-mounts which fit various types of cameras. As I remember, the Nikon's are usually bayonet mounted). Push/Pull the bar on the phototube so that all of the light is going to the camera. Open the camera back and place a small piece of diffuse at the film plane. (A small piece of Fresnel lens also works well). Adjust the height of the phototube until that image is perfectly in focus when the image is in focus for the microscope. From that point on, focusing the microscope should also ensure accurate focus at the film plane, for all magnifications equal to or greater than the objective you used.
For a more detailed discussion as to why all this works, I encourage you to see our book "Optimizing Light Microscopy for Biological and Clinical Laboratories". Details are available at our website: { {http://www.MME-Microscopy.com/education} .
1) Darkfield illumination is interesting because it shows many of the minute details. The preps need to be extremely clean, however, to avoid distraction from dust, etc.
2) Rheinberg illumination is a derivation. We have sets of photographic filters available for minimal cost. Please contact me off line for further information.
3) Hoffman Modulation Contrast might also work, although I have never used it in this particular application.
Details of all these techniques are in "Optimizing Light Microscopy for Biological and Clinical Laboratories" and details for the book are on our website: { {http://www.MME-Microscopy.com/education}
I do have one other contact who is a world-class expert on mites: Mike Huben. I will have to locate his information and send it to you.
Best regards,
At 03:06 PM 8/20/98 MST/MDT, Dr. Mark W. Lund wrote:
} From: Gary M. Easton {geaston-at-ibm.net} To: MSA {microscopy-at-sparc5.microscopy.com} Date: Friday, August 21, 1998 3:24 PMSubject: FREE KEVEX XRAY ANALYZER Hi all, !! FREE FREE FREE !! (I need the warehouse space!) Free to a good caring home - KEVEX Analyst 8000 EDS system(no detector). System is(as far as I've been told) operational. Includes all manuals and SEM interface. You arrange for the packaging and transportation and its yours. FREE. Please reply via email or phone. Gary M. Easton, Pres. SCANNERS CORPORATION Third party SEM service 410-549-3800 x102 gary.easton-at-scannerscorp.com
Please respond directly to the student if you can help him with his question.
Nestor
Dear sir/madam
I have been attempting to submit a question on the internet, but am not having much luck getting it through, please could you forward this question to the correct person.
Name : Alec Higgs School Name : Drakensberg Primary Grade/Level : 6 City : Newcastle State : Kwazulu-Natal Zip : 2940 Country : Republic of South Africa E-Mail address : alech-at-a6.new.iscorltd.co.za
Question :
I would like to know if different races have different shapes in their actual hair strand, Eg. oval or flat or round. (I do not mean curly or straight hair)
Please let me know if this does exist and detail the different races and their different shapes of their hair strands.
unsubscribe Dr. Massimo Catalano Istituto per lo studio di nuovi materiali per l'elettronica del Consiglio Nazionale delle Ricerche CNR-IME Via Arnesano 73100 Lecce - ITALY tel: +39 832 322362 fax: +39 832 325299 email: catalano-at-osfime.unile.it {http://www.ime.le.cnr.it/} http://www.ime.le.cnr.it
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Colleagues } } Please respond directly to the student if you can help him } with his question. } } Nestor } } Dear sir/madam } } I have been attempting to submit a question on the internet, but am } not having much luck getting it through, please could you forward } this question to the correct person. } } Name : Alec Higgs } School Name : Drakensberg Primary } Grade/Level : 6 } City : Newcastle } State : Kwazulu-Natal } Zip : 2940 } Country : Republic of South Africa } E-Mail address : alech-at-a6.new.iscorltd.co.za } } Question : } } I would like to know if different races have different shapes in } their actual hair strand, Eg. oval or flat or round. (I do not mean } curly or straight hair) } } Please let me know if this does exist and detail the different races } and their different shapes of their hair strands. } } Thank you for your response in this regard. } } Alec Higgs
About twenty years ago, I was employed by a company called "The Aerospace Corporation". They had several experimental projects under development. One was "gunshot residue" analysis which is common today. Another was the study of different hair samples. It was hoped that different hair samples could be linked to a given suspect (This was before DNA analysis was available). At the very least, it was hoped to correlate a hair sample to the suspect's race.
In a test, the scientist collected several hair samples from diffrent employees and tested their theory.
The results proved to be embarassing. Out of the dozens of hair samples collected, they concluded that only two hair samples matched. The donors of the two hair samples were the same race or at least related. The reality was that one sample came from a lady of European descent (with undyed blond hair) while the other was a male of African heritage. The program was discontinued.
Oh well, at least the "gunshot residue program" proved sucessful.
Dust mites and other mites can be made more visible (general morphology) by mounting in media with low refracting indice.
Lots of recipes are published: gum acc. to Faure, acc. to Berlese, acc. to Andre, polyvinyl alcohol lacto-phenol, gum dammar dissolved in 2-propanol, gum acc. to Apathy etc... I'll be glad to send you the recipes/protocols/references...
Impregnation in Amman's lacto-phenol and subsequent mounting in glycerin jelly acc. to Kaiser is also a usable method for general morphology.
I have had good results on Demodex foliculorum and Sarcoptes scabiei (an exoparasite in rabbits) with these staining techniques:
1. Staining with Ziehl-Neelsen carbolfuchsine -----------------------------------------------------------------
* Fixation in boiling ethylalcohol 70% the animals died in a perfect extended condition * Flood the object with a few drops of Ziehl-Neelsen carbolfuchsine and warm very gently until steaming * blot very gently dry (without touching the animals!) and differentiate with liquid phenol or acid alcohol (1 ml hydrochloric acid 37% in 100 ml ethylalcohol 70%) * mount in a water based non-acid mountant, or * dehydrate, clear and mount in DPX (numount, cedax...whatever).
2. Staining with pyrogallol --------------------------------------
* Stain the objects in 1% pyrogallol in distilled water or ethylalcohol 70% for about 30 min. * Bring the objects in distilled water or ethylalcohol 70% and let stand in bright daylight for a few hours, the color develops slowly.
When the color is to light, repeat the staining step; when the color is to intense, differentiate in acid alcohol (1 ml hydrochloric acid 37% in 100 ml ethylalcohol 70%)
* mount in a water based non-acid mountant, or * dehydrate, clear and mount in DPX (numount, cedax...whatever).
Staining with a saturated solution of picric acid in clove oil, followed by rinsing in xylene or toluene and mounting in DPX can give sometimes good results: the animals are stained a nice yellow-brown.
It's sometimes possible to stain exoskeletons of mites with Mayers hemalum, Grenachers boraxcarmine or hydrochoric acid carmine.
Yvan Lindekens, Belgium.
---------- } Van: Dr. Mark W. Lund {lundm-at-physc3.byu.edu} } Aan: microscopy-at-sparc5.microscopy.com } CC: lundm-at-physc3.byu.edu } Onderwerp: Stain for dust mites in light microscopy } Datum: donderdag 20 augustus 1998 17:06 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Can anyone recommend a way to stain dust } mites to make them more visible in } a light microscope? } } best regards } mark } }
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We like to inform about the Workshop "Mikromanipulationstechniken in der Mikroskopie " ( Language is German )
{H3} Mittwoch, 18. November {/H3} {HR SIZE=13 WIDTH="100%"} {BR} {BR} {TABLE BORDER COLS=3 WIDTH="100%" BGCOLOR="#FFFFFF" } {TR} {TD} 9.00-15.00 {/TD} {TD} Paktische Übungen an eigenen Präparaten nach vorheriger Absprache {/TD} {TD} {/TD} {/TR} {/TABLE}
{H3} Tagungsort: {/H3} Die Vorträge finden im Konferenzsaal des Hauptgebäudes, die praktischen Übungen im Seminarraum und den Mikroskopielabors des Laborgebäudes Chemie des IPF statt. {H3} Posterbeiträge: {/H3} Im Rahmen des Workshops ist eine Poster-präsentation zu Entwicklungen und Anwendungen von Mikromanipulationstechniken vorgesehen. Zur Festlegung des endgültigen Progamms sollte die Anmeldung der Posterbeiträge, ebenso wie von Kurzvorträgen auf der Tagungsanmeldung bis zum 25.10.1998 erfolgen. {H3} Tagungsgebühren: {/H3} Die Teilnahmegebühren in Höhe von {BR} 350,- DM respektive von 170,- DM für Teilnehmer mit Poster- oder Vortrag (nicht eingeladen) bitten wir unter dem Stichwort „Mikromanipulation“ auf nachfolgend aufgeführte Bankverbindung zu überweisen: {BR} Kto: 0526287200 BLZ: 850 800 00 {BR} Dresdner Bank AG Dresden {H3} Anmeldefristen: {/H3} Um den Workshopcharakter der Veranstaltung zu wahren, ist die Teilnehmerzahl auf maximal 30 Teilnehmer begrenzt. Eine rechtzeitige Anmeldung spätestens bis zum {FONT COLOR="#FF0000"} 25.10.1998 {/FONT} ist unbedingt erforderlich. {H3} Unterbringung: {/H3} Für Zimmerreservierungen wenden Sie sich bitte direkt an die Adressen der der Anmeldung beigefügten Liste von Hotels in unmittelbarer Nachbarschaft zum Institut, zum Bahnhof und zur Innenstadt oder an die Zimmervermittlung „Welcome Tourist“. {BR} {BR} {CENTER} {TABLE BORDER COLS=1 WIDTH="100%" BACKGROUND="10280033.gif" } {TR} {TD} {CENTER} {B} {FONT COLOR="#FF0000"} {FONT SIZE=+3} Versäumen Sie nicht ein Highlight 1998 ! {/FONT} {/FONT} {/B} {/CENTER}
Dear Fellow-listers, We are using the chicken embryo chorioallontoic membrane (CAM) model to study the blood vessel formation and its response to various treatments. I would like to quantify the results by measuring length, density amd branching of blood vessels using image analysis. I wonder if any of you is doing this and would appreciate any experience with image analysis, such as algorithms, type of software best suitable etc. Thank you,
Sarka Lhotak
Hamilton Regional Cancer Centre McMaster University Hamilton, Ontario, Canada
Hi =20 I am trying to inverse the gray levels of an image using the function=20 IpAnam (visilog software). =20 I have used the following=20 IpAnam("imagein";"imageout";{255,0},{0,255},0,255); =20 but this does not inverse the original image.....does anyone know what= =20 the arguments should be? I am afraid I don't quite understand the=20 explanations given in the manual =20 Franck. =20
I am considering selling our H-9000 HREM. This is a serious HREM with better than 1.8 Angstroms resolution, and comes with a Gatan TV camera. It has been (and still is) under maintenance contract. Some general information is available at:
http://www.numis.nwu.edu/internet/hrem.html
and contact me for further information.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
ELECTRON MICROSCOPY TECHNICIAN The Integrated Microscopy Core, Department of Cell Biology, Baylor = College of Medicine has an immediate full-time opening for an electron = microscopy technician. The Integrated Microscopy Core is a busy, = state-of-the-art facility with 2 TEMs, deconvolution, laser scanning = confocal and 2 CCD-based fluorescent microscopes. The applicant should = have at least 1-2 years experience in all aspects of sample preparation = for biological TEM including fixation, embedding, ultrathin sectioning = and staining of tissue samples and cell monolayers. The applicant = should have darkroom experience and experience in the operation of TEMs. = Immunolabelling experience is desirable but not essential. Other = duties include preparation of solutions, embedding media and the = maintaining of records. The position offers opportunities for training = in advanced light microscopy techniques, including fluorescence, laser = scanning confocal and deconvolution microscopy. The position requires a = minimum of a Bachelors degree and will start as=20
a Lab Technician II; salary range is low 20's, commensurate with = experience, and includes the standard Baylor benefits package.
Send CV and letter of research/technical interests to:
Hank Adams Laboratory Manager Integrated Microscopy Core Department of Cell Biology Baylor College of Medicine One Baylor Plaza Houston, TX 77030 Email submissions to: hpadams-at-bcm.tmc.edu Fax submissions to: 713 790 0545
Baylor College of Medicine is an Equal Opportunity, Affirmative Action = and Equal Access Employer.
I am about to compile a table of units of measure including SI, other metric, and customary units, with emphasis of those units used in biomedical sciences. The goal of this work is not just to enumerate all units (which is barely possible) but to list only the terminal symbols used as units in algebraical expressions of units. These unit atoms are associated with a precise semantics expressed by means of linear algebra.
Here I am interested in the unit symbols "low power field" (LPF) and "high power field" (HPF). These are commonly used in clinical medicine as a unit to measure the size of the reference system when counting material of interest in microsocpy. For example, erythrocytes, leucocytes in a unrine sediment can be reported as 10/HPF (10 per high power field).
Clearly, LPF and HPF are no exact units but rather qualifiers of semiquantitative observations. However, clinically, those observations are sometimes treated like hard numerical observations, including such uses as criteria for eligibility in clinical trials.
For inclusion into the table of units there are two options. Treat LPF and HPF independently as dimensionless units with value 1, which is the last resort option for all those difficult to compare units. However, this looses even the obvious relationship between LPF and HPF.
The other extreme is to treat LPF and HPF as proper volumes of fluid that overwiewed in the microsocope. This is probably hard to estimate and too variable.
Therefore, I could imagine treating the LPF and HPF as actual areas (measured in square-millimer) so that the per-LPF and per-HPF quantities would be convertible to areic numbers. While this is still incompareable to most other units in the system, it does at least establish a relationship between LPF and HPF.
I need the help of some expert in microscopy to give me an estimate of the typical view area in those magnifications typically called "low power" and "high power". It would also be of interest to know whether there are considerable subjective variations to those observations (i.e. what microscope is used? Monocular/binocular? Does the observer wear glasses? If so, does (s)he use apropriate oculars or normal oculars?) and to have an estimation of the error that results from those variations.
Any comment or advice is highly appreciated.
best regards -Gunther Schadow
Gunther Schadow ----------------------------------- http://aurora.rg.iupui.edu Regenstrief Institute for Health Care 1001 W 10th Street RG5, Indianapolis IN 46202, Phone: (317) 630 7960 schadow-at-aurora.rg.iupui.edu ---------------------- #include {usual/disclaimer}
Thank you very much for the reply on storing the floating sections question. There seem to be few options, all within the same idea. In fact I seem to have hard time deciding which one to use. If any of you interested, I will forward you the replies. Hope to be of help in the near future, Lilith ---------------------------------- Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
Can anybody tell me if the wax on the plant surfaces (aerial organs) does autofluoresce? Or, better off, send me to a reference. Sections of leek leaves show light green autofluorescence (filter BV-2A) at the surface of the epidermal cells, where the crystalline epicuticular wax is deposited. The cuticle may autofluoresce too, but I'm not sure. Thank you very much.
Camelia G.-A. Maier Postdoctoral Fellow The Samuel R. Noble Foundation Plant Biology Division Ardmore, Oklahoma
I will have to check on the exact procedure you are discussing but there are some very specific answers to your later questions:
1. The field of view for any objective can be calculated by dividing the field number by the total objective magnification.
a. The field number is usually engraved on the eyepiece and is typically something between 18 and 25. It refers to the diameter, in mm, across a small baffle or ridge which is placed anywhere between the mid line and lower 1/4 of the eyepiece. If you pull the eyepiece out, turn it upside down, then gently run a finger inside the barrel, you can usually feel it. In some types of eyepieces, (Huygen), it is mounted between the top and bottom (field) lens but you can usually still see it.
b. The total objective magnification is the magnification of the objective times the magnification of any other optics (tube lens, optivar or intermediate magnifier) up to but not including the eyepiece.
Ex:
For an 18mm field of view eyepiece and a standard 10x objective (most likely the one indicated by your LPF), the field of view would be 18mm/10. The resulting field of view is 1.8 mm or 1800 micrometers.
Ex II:
For a 20 mm field of view eyepiece and a 100x objective (most likely the one indicated by your HPF), the field of view would be 20 mm/100. The resulting field of view is 0.2mm or 200 micrometers.
If you used a 2x intermediate magnifier in this last example, it would drop the field of view to 100 micrometers.
2. This relationship holds irrespective of the brand of microscope used, of monocular/binocular, and with or without glasses.
For further information on these and related topics, might I suggest "Optimizing Light Microscopy for Biological and Clnical Laboratories". Details are available at our website. The book should be available locally through dealers who carry products from Structure Probe, Inc. If you cannot find it there, ordering information is also available on the website.
thanks for you quick and detailed answer! This helps a lot. When you say ``18 mm field'' or ``20 mm field'' in your example I assume that you speak of the diameter of a circular area, is that correct? It isn't by chance the radius?
} From what you say, I unfortunately have to conclude that the size of the so-called LPF and HPF depends on the type of eyepiece used by the observer. And the area seems to vary considerably as I calculated in the following table:
============================================= 18 FIELD NUMBER 25 --------------------------------------------- LPF (x10) 2.544 mm2 4.909 mm2 HPF (x100) 0.02545 mm2 0.04908 mm2 =============================================
This is a relative error of 63 % which is quite bad. When LPF and HPF are defined in terms of length (i.e. diameter) rather than area, I can press the error down to 32 % which is still way too high. It seems to me that really the only possible thing is to tell LPF from HPF by a factor of 100. So I could define LPF as 1 and HPF as 100. Any conversion to an actual area, let even volume, would be unreasonable due to this huge error that is inherent in those units.
} I will have to check on the exact procedure you are discussing but ...
So there is some last chance that LPF and HLP may be defined more exactly? Are there any recommendation by the Microscopy Society of America or some standards body concerning the use of LPF and HPF? Any standardizations or deprecations?
many thanks, -Gunther
Gunther Schadow ----------------------------------- http://aurora.rg.iupui.edu Regenstrief Institute for Health Care 1001 W 10th Street RG5, Indianapolis IN 46202, Phone: (317) 630 7960 schadow-at-aurora.rg.iupui.edu ---------------------- #include {usual/disclaimer}
Alec, I cannot answer your question...I am just awfully curious how you expect to define "race". Did you know that the differences within a so-called race, however you like to define it, is not significantly different from the differences between them. This is just the law of statistics that does not allow for such a differentiation, it is just in the prejudiced mind.
A fellow South African Quirina
} Dear sir/madam } } I have been attempting to submit a question on the internet, but am } not having much luck getting it through, please could you forward } this question to the correct person. } } Name : Alec Higgs } School Name : Drakensberg Primary } Grade/Level : 6 } City : Newcastle } State : Kwazulu-Natal } Zip : 2940 } Country : Republic of South Africa } E-Mail address : alech-at-a6.new.iscorltd.co.za } } Question : } } I would like to know if different races have different shapes in } their actual hair strand, Eg. oval or flat or round. (I do not mean } curly or straight hair) } } Please let me know if this does exist and detail the different races } and their different shapes of their hair strands. } } Thank you for your response in this regard. } } Alec Higgs
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
I'm trying to reach through e-mail Sales & Marketing Department of Energy Beam Sciences Inc. As so far without success. Actually, I'm not sure if it's the address or maybe our server's problem. Could anybody tell me if the address I'm using is the right one. It is: 75767,640-at-CMPUSERVE.COM.
Thanks in advance
Witold Zielinski Warsaw University of Technology Faculty of Materials Science and Eng. Narbutta 85 02 524 Warsaw Poland
A quick thank you to everyone that responded to the difficult task of cleaning oils... from powder metal samples. Many of the ideas have been tried in the past although with inconsistent results. The plasma etching method sounds promising although I have yet to try it as I am waiting for more samples. I am curious about the depth of cleaning while using this technique as the samples are usually a few cm thick .?. A quick run through will tell soon enough.
Thanks again. Wayne wengland-at-ortech.on.ca
XX
I just read your message and though I am no expert, a thought came to mind. Have you considered cleaning the metal powder in an oxygen plasma? I suspect that if it were to work you would probably have to clean as much of the oil off as possible by your current methods and try the plasma as the very last step. It might oxidize the surface of your metals, too. A question - when you heated the powder was that done in a vacuum or in air? We often heat our TEM samples in a (very good) vacuum to drive off hydrocarbons and have had good success.
I hope this is of some help.
xx
How about using an ESEM? It doesnt really care if there is some oil left, wash the stuff in a solvent and then put it in the ESEM. You can come and use ours if you want.
xx
Wayne, Do you have the capability to put the parts in a solvent reflux unit to provide a continuous wash over a 24 hr period? This should work especialy if it is set up to alloy the recondenced solvent to drip on the top of the part.
xx
Filter them onto a suitable membramne (i.e. Millipore) filter and flush it with a solvent for the oil. You'll have to choose a filter material that won't be dissolved by the solvent. If you precoat the filter with Au/Pd you shouldn't have any charging problems.
xx
Oil impregnated powdered metal is usually cleaned by extracting in petroleum ether. The process is to soak in ether, weigh the material, soak in ether and reweigh. Repeat the process until the weight does not change. Soak times may takes hours or even a few days.
xx
I was wondering what types of metals you were trying to clean? Also, when you heated the powder, did you heat it in a specific atmosphere of just an open air type furnace. And did you use any soaps or something that will 'capture' the oil molecules??
xx
Some Ideas for a very difficult problem. We have used a vacuum and clean in solvent in an ultrasonic cleaner. This process may require multiple repetitive steps.Running the parts through a degreaser if one is available may help. A degreaser that uses heat and vapor may be the way to go. If you have small parts a cold finger over an boiling acetone with a still the drips the condensed acetone on the part may be effective also. (TEM replica removal set is what I am trying to describe.) You may want to set a highly absorbent paper near where you want to look to help wick the oil way form the pores or crack(s). If you place the paper in a vacuum you may need to dry or out gas the paper first. If you are looking at oil impregnated parts such as bearings good luck. (I use Simple Green Cleaner, but this is not an endorsement of this product.) There are other water based solvents that work just as well or maybe better. Petroleum solvents work also but are harder to dispose of.
ASM in their metals handbook on metallography also has technique for cleaning samples. One old time engineer described this cleaning process as "worring over the part."
xx
Have you considered plasma cleaning? We market a system that was designed specifically for cleaning organic contamination from EM specimens. It utilizes a low energy, inductively coupled plasma to selectively remove the contamination without altering the specimen material. The process incorporates an oxygen plasma in which the disassociated oxygen created by the plasma chemically combines with the carbonaceous material. The resultant is a combination of CO, CO2 and H2O.
In our own laboratory when we have had small ball bearings and ball bearning raceways to be examined, and we don't want to use solvents, etc. out of fear of losing corrosion product or other materials from the analysis, we have used quite successfully our own SPI Plasma Prep II Plasma Etcher.
If you are not familiar with it, you can find it on our website URL http://www.2spi.com/catalog/instruments/etchers1.html
I toyed with the idea of making a public posting on the listserver but feared having it look "too commercial". However, if you should post any kind of summary, you certainly have my permission to post this information.
xx
In terms of actual metal powder used in powder metallurgical operations, when we receive them in the laboratory, they are often times containing something sorbed to their surfaces, but a quick exposure to an oxygen plasma within a few seconds etches away this layer resulting in a far better end result in the micrographs.
xx
I have been playing around with sovelnts (diff pump stack revival) Carbon tertracloride CCl4 fumes are brilliant. It boils at 76.54 deg Centegrade. Be sure that you work in a fumehood! Is used as a degraser at Cr and Ni coating facilitys.
xx
While I have not done SEM on powdered metals, I do have a bit of experience in cleaning the packing oils as I used to use powdered metals as reagents for high-temperature synthetic chemistry of intermetallics back in graduate school. I was extremely concerned about having no residual oil and as little surface oxidation as possible- also, several of the powdered metals I used were very reactive (in terms of reacting with air to form oxides). So, for your problem, it depends a lot on what the metal is (how fast it reacts with air) and how clean you want the surface to be. For extremely clean surfaces, I had friends in the physics department that would prepare a freshly cleaved surface from a bulk sample under the UHV conditions in their PES system. Techniques that I used involved repeatedly washing the powder in an appropriate solvent, using either filter paper on a buchner funnel or with special glassware having porous-glass filter-frits. For the very reactive metals (some of the lanthanides and occaisionally potassium or rubidium) I would work either on a schlenk line or in a nitrogen purged glove box using freshly distilled (extra-dry) solvents (usually pentanes, or hexanes or ether- depends on the oil- I'm not seure what is best for transmission fluid; I don't know what that is actually). For less reactive powders I could do a decent job with with solvents right out of the bottle working in air (in a fume hood)- then backfilling the container with dry N2 for storage. If the powder is very easily oxidized, you also have the problem of getting from an inert atmosphere to the SEM- for this you might get creative with a plastic bag that can be back-filled (or flushed several times) with dry nitrogen. There's lots of ways to do this kind of stuff on the cheap (if you aren't going to be doing it too often). You can find a lot of explanations and diagrams etc in a good inorganic (or organic for that matter) chemistry text book. The degree of care needed will be determined rather simply by the reactivity of the powder and the cleanliness that you require.
One disclamer is that several metal powders that are shipped under oil are done so as they react very violently with air or water- so be careful- (maybe you kno0w all this, but I don't want to be responsible) my advice is to check your library or local university bookstore for a good inorganic chemistry text so that you can know if their are potential hazards (MSDS can also be helpful- but they aren't always so well written).
Could someone responsible for submissions to the magazine Microscopy Today please reply to my personal address? A while back I was asked to contribute an article by someone whose e-mail address is no longer valid - I wish to find out if the contribution is still desired. Rob Palmer Director - Biofilm Imaging Facility CEB/UT
Witold Zielinski asked: } I'm trying to reach through e-mail Sales & Marketing Department of Energy Beam Sciences Inc. } As so far without success. Actually, I'm not sure if it's the address or maybe our server's } problem. Could anybody tell me if the address I'm using is the right one. It is: } 75767,640-at-CMPUSERVE.COM.
The Compuserve account has been closed. Our general e-mail address is ebs-at-ebsciences.com
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
Our Epson FX-85 dot matrix printer which came with our System 4-plus EDS in 1986 finally died. Attempts to send the print command to later Epson printers (FX-86E) have not worked. Is there a fix or upgrade that has been successful by users from the RT-11 system? Thanks in advance.
} Can anybody tell me if the wax on the plant surfaces (aerial organs) does
} autofluoresce? Or, better off, send me to a reference. Sections of leek
} leaves show light green autofluorescence (filter BV-2A) at the surface of
} the epidermal cells, where the crystalline epicuticular wax is deposited.
} The cuticle may autofluoresce too, but I'm not sure.
} Thank you very much.
}
} Camelia G.-A. Maier
} Postdoctoral Fellow
} The Samuel R. Noble Foundation
} Plant Biology Division
} Ardmore, Oklahoma
}
}
Camelia,
One answer to your questions lays in the chemical structure of the wax. If it has conjugated bonds (alternating double and single) which allow mobility for pi bonding electrons, then it is likely to fluoresce. The more conjugation, the further the fluorescence will be to the red end of the spectrum.
One caution: chlorophyll is autofluorescent (green excitation, red emission) so it may be difficult to distinguish the wax if it is sitting on a leaf surface. You might try cross-sectioning the leaf and looking at the thin cross-section.
Re: defining LPF and HLP: yes. We have an expert Med Tech available but she is on vacation at the moment. I'll try to get you that info within the next week or so.
Re: the numbers 18 and 25. These are, indeed, the diameter across the open space defined by the baffle mentioned in the earlier posting.
I don't think defining these terms as arbitrary values of 1 and 100 does anyone any good. I think you need to be honest with what the numbers really represent. Since the American Optical Microstar 110 was the "industry standard" for so long. Unfortunately, those archives are packed up at the moment, but I think that the field number on the microscope was either 18 mm or 18.5mm. (The old American Optical line is now part of Leica so you might try calling their home office in Deerfield, IL).
concerning toxicity and waste disposal of fixatives, resins and other EM reagents I am looking for the print "Safety Chart, Chemicals in Electron Microscopy" from EMscope Laboratories Ltd., Kingsnorth Industrial Estate, Ashford, Kent. Has anybody the full adress or faxnumber of EMscope or could sent me a copy of the print? Furthermore I would be interested to know about the national guidelines for EM waste disposal, e.g. in the US and UK. Any comments are welcome!
Steven E. Slap, Vice-President P.O. Box 468 11 Bowles Road Agawam, MA 01001-0468 USA Phone: (413)786-9322 Phone:(800)992-9037 Fax: (413)789-2786
Click here to send mail: ebs-at-ebsciences.com ^^^^^^^^^^^^^^^^^^This may be what you're looking for
Tungsten filaments, Denka LaB6 and TFE cathodes; laboratory microwave processors; JB-4 microtome, triangular and Ralph knifemakers; Vibratome and MicroCut vibrating blade microtomes; Polaron sputter coaters, carbon coaters, critical point dryers, plasma ashers, vacuum evaporators and SEM cryo-preparation system; x-ray microanalysis standards; EM and histology embedding kits; EM automatic film processor.
regards -Gunther
Gunther Schadow ----------------------------------- http://aurora.rg.iupui.edu Regenstrief Institute for Health Care 1001 W 10th Street RG5, Indianapolis IN 46202, Phone: (317) 630 7960 schadow-at-aurora.rg.iupui.edu ---------------------- #include {usual/disclaimer}
Email, WWW sites, Phone, Product information, and address for hundreds of vendors are listed in the Directory of Microscopy Vendors at http://www.mwrn.com/vendors/directory.htm
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Susanne
At 07:23 AM 8/25/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi Camelia, my experience is that epicuticular waxes do fluoresce but... Are the sections you're working on fixed? Glutaraldehyde in tissues that are resin embedded will result in a strong greenish fluorescence that is actually really welcome in some anatomical studies. You would have to look at some fresh tissue as a control. John
} } Hello all } } Can anybody tell me if the wax on the plant surfaces (aerial organs) does } autofluoresce? Or, better off, send me to a reference. Sections of leek } leaves show light green autofluorescence (filter BV-2A) at the surface of } the epidermal cells, where the crystalline epicuticular wax is deposited. } The cuticle may autofluoresce too, but I'm not sure. } Thank you very much. } } Camelia G.-A. Maier } Postdoctoral Fellow } The Samuel R. Noble Foundation } Plant Biology Division } Ardmore, Oklahoma
________________________ C. John Runions Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
We are a world-wide mineral company providing both natural and synthetic = mineral products to a number of industries including paper, plastic, = refractory, steel and food.
Title of Posted Position : Analytical Investigator Location: Easton, PA Division: Specialty Minerals, Inc. Department: Analytical Services
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All inquiries should be directed to the Human Resources Department:
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{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} We are a world-wide mineral company = providing=20 both natural and synthetic mineral products to a number of industries = including=20 paper, plastic, refractory, steel and food. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Title of Posted Position=20 : = Analytical=20 Investigator {/FONT} {/DIV} {DIV} {FONT=20 size=3D2} Location: &= nbsp; &n= bsp; &nb= sp;=20 Easton, PA {/FONT} {/DIV} {DIV} {FONT=20 size=3D2} Division: &= nbsp; &n= bsp; &nb= sp; =20 Specialty Minerals, Inc. {/FONT} {/DIV} {DIV} {FONT=20 size=3D2} Department: = ; = =20 Analytical Services {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Typical Duties: {/FONT} {/DIV} {DIV} {FONT size=3D2} Under minimal supervision, The Analytical = Investigator=20 (Microscopy) is responsible for performing the chemical and microscopy=20 investigations of samples, maintaining technical mastery and awareness = of the=20 state-of-the-art for areas of responsibility, performing administrative = duties,=20 and actively assisting the Analytical Services Group achieve its = Objectives and=20 fulfill its Mission. Primary duty is customer oriented problem solving = through=20 the use of optical and electron microscopy and microchemical analysis in = a team=20 environment. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Qualifications: {/FONT} {/DIV} {DIV} {FONT size=3D2} Incumbent should possess a Bachelor's Degree in the = physical=20 sciences or engineering, preferably in chemistry or mineralogy and have = a=20 minimum of six years of chemical analysis or microscopy experience, = preferably=20 in a service environment. The Analytical Investigator performs work of a = varied=20 nature and is responsible for making some of and implementing most of = the=20 decisions relevant to the areas of responsibility. Must have excellent = oral and=20 written communication skills, computer skills, team skills, ability to = interact=20 with a variety of people, and the ability to operate analytical = instrumentation=20 and to manipulate equipment and materials weighing 30-50 pounds. = Previous=20 analytical and LIMS experience desireable. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} All inquiries should be directed to the Human = Resources=20 Department: {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Gary Duckwall, HR Manager {/FONT} {/DIV} {DIV} {FONT size=3D2} Specialty Minerals, Inc. {/FONT} {/DIV} {DIV} {FONT size=3D2} 640 N. 13th Street {/FONT} {/DIV} {DIV} {FONT size=3D2} Easton, PA 18042 {/FONT} {/DIV} {/BODY} {/HTML}
I am at the University of Massachusetts Lowell. I have a VG ESCALAB II. The hardcopy output of the PDP11/83 is connected to an (obsolete) HP 7550A multipen plotter. After pen retrieval the pen comes down touches the paper, lifts back up and goes through its routine, for example, drawing the curve, but it is not in contact with the paper so it does not plot. The HP instructional book does not discuss this issue. Is it a software or a harware problem?? I need some direction. Thanks Dan Oblas
Hi there folks, I think the problem may be hardware if it does that with every color pen. Now there were different length pens at one tie also. Are you sure of what you have loaded? Just a thought out of a foggy ex-hp resellers engineers brain! Ed Sharpe
Microscopy Vendors Database at http://www.kaker.com is currently accessible only under IP address http://207.137.96.185/mvd/vendors.html, because we are in process of replacing current ISP with new one.
I am looking for THIN-pointed (not welded-pointed) filaments for our Siemens Elmiskop 102 electron microscope. Does anybody have some information about this article ?
Thanks in advance
Philippe Drouillon Solvay Research and Technology Electron Microscopy and Image Analysis Rue de Ransbeek, 310 B-1120 Brussels (Belgium) phone : (00 32) 2 264 24 47 fax : (00 32) 2 264 20 55 mailto:philippe.drouillon-at-solvay.com
------------------------------------------------------ Dr. B=E9la P=E9cz Research Institute for Technical Physics and Matl. Sci. H-1525 Budapest, POBox 49 Hungary phone: 36-1-395-9240 fax: 36-1-395-9284 E-Mail: pecz-at-mfa.kfki.hu ------------------------------------------------------
Philippe Drouillon asked: } I am looking for THIN-pointed (not welded-pointed) filaments for our } Siemens Elmiskop 102 electron microscope. } Does anybody have some information about this article ?
We (Energy Beam Sciences) have manufactured a range of pointed filaments for almost 30 years. There are several styles available, and they can be mounted on any filament base. Please contact me back-channel for additional information, or visit our website (address in my .sig, below).
I am looking for a distributer in Germany/Europe for antibodies from Miles Laboratories Inc. I even cant get a valid (www)-address of the mother company.
i am trying to determine the grain size of aisi 1050 that has been spheroidized is there an etch that would let me see the grains and not the carbon phase?
I am looking for a pan keratin antibody that works at the EM level on cultured keratinocytes that have been fixed in Zambonis (para-picric acid), and embedded in LRWhite.
I need to lable the cyto-skeleton and also our protien of interest. Does anyone know of one that works?
Can anyone help me with an address, web-site, or phone number for RMC in the US? We are looking at purchasing an ultramicrotome with cryo-adaptor. Anyone have any preferences between RMC's and Leica's models? Thanks Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219 (214) 559 7744
Professional electron microscopist with a minimum of 5 years experience to manage the daily activities of the EM Facility at the University of Missouri-Kansas City School of Dentistry. Duties would include active participation in ongoing research, troubleshooting, scheduling, and training on facility instruments: Philips CM12 STEM with new Princeton Gamma Tech EDS system; Philips 515 SEM with new Princeton Gamma Tech EDS system and Robinson backscatter electron detector; and recently-installed Philips Environmental SEM equipped with Schottky field emission source, gaseous electron detector and new Princeton Gamma Tech EDS system. Applicant should have operational and maintenance experience with transmission and scanning electron microscopes. Additionally knowledge of energy dispersive spectroscopy and specimen preparatory techniques is required. The position is affiliated with the Department of Oral Biology at UMKC School of Dentistry. Salary is commensurate with level of education and experience. Please respond by E mail to "eickd-at-umkc.edu" with resume.
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} Posted-Date: Tue, 25 Aug 1998 16:29:53 +0200 } X-Sender: neubohn-at-mendel.ipk-gatersleben.de } Date: Tue, 25 Aug 1998 16:29:53 +0200 } To: microscopy-at-sparc5.microscopy.com } From: Birgit Neubohn {neubohn-at-IPK-Gatersleben.de} } Subject: toxicity of EM reagents } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Given Birgit requests guidelines and comments, this may be of interest. It was originally sent to MICROSCOPY TODAY, but was not published, presumably because the topic was viewed as parochial. ***** Uranium compounds, especially uranyl acetate, have been widely and routinely used as transmission electron microscopy (TEM) contrast stains for biological materials since 1958 (1,2). Those who do TEM of biologicals use small quantities of uranyl acetate, nitrate, formate, sulfate and perhaps other uranuim compounds almost daily and therefore keep inventories of these salts and their solutions.
In the 1980's growing concerns about medical and research wastes entering regional dump sites prompted state radiation officials in Oregon to begin tightening the regulations for monitoring and controlling all radioactive substances, including the uranium compounds commonly used in processing biological specimens for TEM. Oregon has come to be a state with high levels of awareness about environmental and safety issues and is often at the forefront of regulatory and management trends in these areas. What happens in Oregon may, therefore, frequently serve as both a harbinger of changing attitudes and a model for standards which are evolving elsewhere. Messages seen in electronic news groups and conversations with colleagues suggest that other states may have also addressed, or may plan to address, the purchase, storage, distribution, use, and disposal of uranium compounds used for staining biological preparations for TEM.
Radiation-producing machines and radioisotopes used and stored at locations under State of Oregon jurisdiction are subject to provisions of the Oregon Rules for Control of Radiation, set forth by the Radiation Protection Services, Health Division, Oregon State Department of Human Resources. In addition, radioactive material transport, storage, and disposal must comply with rules issued by the Oregon Department of Energy, Oregon Department of Environmental Quality, and applicable federal agencies such as the U.S. Nuclear Regulatory Commission and U.S. Environmental Protection Agency. Regulations covering naturally occurring radioactive materials (NORM), a class of materials which includes uranium compounds used as stains in EM, have been in place in Oregon for several years (3). When these regulations were established, Oregon State University (OSU) undertook regulatory investigations to determine if use of small quantities of NORM could, or should, fall within the guidelines for exemption allowed under these regulations.
In 1992 the investigations were completed and a process to include uranium compounds used as stains for electron microscopy in the state's radiation regulatory and safety program was begun. Hence, it was at this time that our use of uranium-based compounds was first called into question. There are may isotopes of uranium. Depending on what mix of these one may be dealing with, various levels of alpha, beta, and gamma radiation can result from their radioactive decay.
On the OSU campus, the radiation safety program is managed by a radiation safety officer responsible to university administration and a University Radiation Safety Committee. The radiation safety officer supervises radiation safety training programs, use, storage, disposal and licensing matters, and coordinates functions of a faculty committee which establishes local policy and reviews safety and compliance matters.
Prior to 1992, neither facility magagement nor our radiation safety officials felt the small amount ( {25 grams with activity {10 microcuries) of NORM uranium compounds on hand in the facility, and their limited methods of use, posed sufficient hazard to warrant regulatory action.
Campus radiation safety personnel have always been understanding, knowledgeable, and non- adversarial in carrying out their responsibilities, but as the state mandates for stricter control were imposed, neither facility management nor the radiation safety officer were sure how best to proceed, with minimal adverse impact, toward compliance. The EM Facility at OSU is a service laboratory: several dozen students, technicians, and faculty use the facility and its supplies, and uranium-stained tissue embedments and grids are typically dispersed into the possession of facility clients. If materials that contained uranium were used by large numbers of people or removed from the "authorized" facility, could we allow these use and dispersal practices and still comply with the spirit and intent of the stricter regulations? The concerns went beyond possible health effects from radioactivity to include possible health effects related to more conventional chemical toxicity issues and the possibilities for contamination effects where inadvertent disperal might result in inaccuracies in low-level radiation monitoring activities.
We began by discussing and arriving at a mutual understanding of the regulatory needs and concerns and the complications that altered protocols and compliance-management would entail on both the EM facility and the radiation safety program. We next undertook to quantify the hazard potential. Our small stock of NORM uranium compounds in powder form were identified, inventoried, weighed and surveyed for radiation levels within their containers (bottles) and exterior to the sealed containers. Two dilute uranyl acetate solutions were likewise identified, inventoried, and quantified. In addition, we processed an assortment of plant, animal, and microbial specimens through standard protocols and submitted for analysis samples of the tissues, pellets, and the used and unused processing solutions before and after uranyl compound staining. We also provided samples of the plastic embedded tissues and pellets, as well as sections on copper TEM grids cut from these embeddments.
One interesting complication in quantifying the hazard arose early in the process. Every other campus user of radioisotopes was using these materials as tracers. These users were concerned with specific activity. As a consequence, their inventories were quantified and managed in protocols in terms of microcuries. For electron microscopy, we were concerned only with the electron scattering potential of the uranium atoms. We quantified our inventories and formulated our solutions on the basis of weights and volumes. A calculation of 0.33 microcurie/gram of U-238 was made to convert uranium compound supplies from weight to specific activity units for inclusion under the hazard assessment required by the stricter regulations.
Radiation levels (millirem/hr) were measured from open and closed containers of NORM radiation compound powders and solutions. Specific activity values associated with these measurements were then calculated from the specific activity of depleted uranium and the weight percent of uranium in the compounds. Working strength uranyl acetate solutions, nominally 1% w/v, gave specific activity measurments below 1x10-3 microcurie.
A variation of neutron activation analysis, gamma-ray spectroscopy using a germanium/ lithium detector, was required to detect uranium in processed tissues and pellets, where uranium levels in the 0.8 to 0.9 ppm range were reported. To assure a high level of accuracy, the analysis equipment was calibrated a number of times, and the results from samples submitted in December 1993 were not obtained until May 1994. The presence of uranium in stained materials on grids was below the detection limit of the analytical equipment and procedures used.
The final result of the quantitative and regulatory processes produced the development and implementation of a policy by which the Radiation Safety Office and EM Facility agreed to cooperate to enforce defined procedures. These are the central features of this policy.
A) Persons using uranium compounds in the EM Facility take a four hour radiation safety training class from our campus radiation safety officer. They then receive authorization to use NORM uranium compounds as stains in the facility. Individuals who want to purchase and maintain their own stocks of uranium-based staining compounds at other locations must obtain a radiation use authorization (RUA) permit for those locations from the campus Radiation Safety Office.
B) After use authorization is granted, users may obtain and manage their own stocks of uranium compounds or may use uranium stain solutions in the EM Facility. Uranium salts and stain solutions provided for use by and in the EM Facility may not be taken to other locations.
C) Persons using uranium-based stains in the facility are personally responsible for complying with all requirements for use, clean-up, in-lab disposal, and radiation monitoring. The EM Facility provides radiation safety and monitoring, clean-up, and disposal materials for client use.
D) The EM Facility manager maintains and documents the status of the uranium compound and radiation safety materials inventory, monitors for indications of radiation contamination or unsafe procedures, and attends to the safe and timely disposal of accumulating radioactive wastes.
E) Specimens which have been stained with any uranium compound(s) for EM and subsequently either solvent-rinsed, plastic embedded, or put on grids may be removed from the EM Facility for archiving or futher processing at other locations as long as uranium concentration gives a specific activity below 0.05 microcurie/gram.
This policy took effect 01 January 1994. Despite a small burden of additional managerial tasks and the minor inconveniences of more record-keeping chores, the policy has, to date, worked well.
Two additional points should be made. Authorization of NORM use in Oregon, and presumably elsewhere, is rather complex. Under Oregon regulations there is still some exemptions allowed for specific NORM uses, materials, concentrations, and amounts. Users of uranium compounds should investigate their specific situation, giving consideration to ALL uses of NORM at their organizations, not only the use of uranium for EM stains.
Finally, it should be noted that a particular problem is associated with the disposal of uranyl nitrate waste. This compound is a radioactive substance, a nitrate, and an oxidizer, and therefore is classified as a mixed radioactive and chemcial hazard.
1) Watson, M.L. (1958). J. Biophy. Biochem. Cytol. 4, 475. 2) Swift, H., Rasch, E. (1958). Sci. Inst. News 3, 1. 3) Radiation Safety Manual, Oregon State University
Al Soeldner Lab Manager EM Facility Oregon State University Corvallis, OR 97331-2902
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I have a user who wants to try to capture a specific virus from a prep contaminated with TMV. I believe there are methods involving placing a layer of antibody against a coat protein from the virus you want to capture on a grid. Then you float the grid on a droplet of the virus prep, wash off any unbound particles and negative stain. Hopefully this would leave only the desired virus on the grid. Does anyone use this technique and/or have a more detailed procedure?
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
by imo15.mx.aol.com (IMOv14_b1.1) id NRBOa20769 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 26 Aug 1998 17:47:56 -0400 (EDT) Message-ID: {b475c5c4.35e4828d-at-aol.com}
need micro manipulation apparatus for use on light microscope. please contact ed sharpe thanks
We have a Polaron diode sputter coater we use exclusively for gold coating for conventional 20 kV tungsten emitter SEM.
Lately the coatings being applied seem comparatively thin - instead of a shiny metallic gold coloured coat we get a semitransparent dark gray coat.
But we are coating using the same parameters as ever : 50 mA -at- 1.4 kV with 0.2 torr of argon.
This occurs even with plain glass specimens which should not be contributing any unwanted vapours.
I have checked out the pumping system and the argon gas bottle is labelled "high purity argon" and is quite new.
I would welcome suggestions as to the source of our problem.
Mel Dickson
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
} Dear sir/madam } } I have been attempting to submit a question on the internet, but am } not having much luck getting it through, please could you forward } this question to the correct person.
} } Name : Alec Higgs } School Name : Drakensberg Primary } Grade/Level : 6 } City : Newcastle } State : Kwazulu-Natal } Zip : 2940 } Country : Republic of South Africa } E-Mail address : alech-at-a6.new.iscorltd.co.za
} } Question : } } I would like to know if different races have different shapes in } their actual hair strand, Eg. oval or flat or round. (I do not mean } curly or straight hair) } } Please let me know if this does exist and detail the different races } and their different shapes of their hair strands. } } Thank you for your response in this regard. } } Alec Higgs
My appologies for the lateness in this reply, I have been out of the country. As an old and still practicing classical hair comparison person my views may be seen by many as biased.
The simple answer is that there has been found a relationship between cross sectional shape and racial characteristics. Flat being associated with the Negroid race, oval with the Caucasion race, and round with Mongoloid.
To quote from "MICROSCOPY OF HAIR, A Practical Guide and Manual" put out by the FBI:
"A human hair may be classified according to its racial characteristics as being Caucasian, Negrod or Mongoloid origin. In some instances, the racial characteristics exhibitied by the hair specimine may not be clearly defined indicting the sounce of the particular hair may be of mixed racial origin."
and
"Again, it is pointed out thet even when racial characteristics are not clearly defined, it is significant when these characteristics are consistent between the hair in questiion and the hair of known origin."
Okay, probably more than you wanted to know. Good luck.
Shalom from Jerusalem, Azriel +++++++++++++++++++++++++++++++++++++++++++++++++++++ Major Azriel Gorski, Head Fibers and Polymers Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem, ISRAEL
azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739
What lies behind us and what lies before us are tiny matters compared to what lies within us. Ralph Waldo Emerson ++++++++++++++++++++++++++++++++++++++++++++++++++++
I am interested in micro-manipulation (e.g. handling, applying external forces, injection................) of structural elements. Any information on the issues mentioned below would be of interest:
* instrumentations/instrument configurations * colleagues with expertise in this area * workshops, courses, conferences * manufacturers of instrumentation * handbooks, literature references
Please reply to:
Marcel Paques Wagenigen Centre for Food Sciences P.O.Box 20, 6710 BA Ede telephone: (31) 318 659690 telefax (31) 318 650400 email: paques-at-nizo.nl
I am posting this query on behalf of a colleague. He wishes to locate a supplier of microscope slides which have fluorescent beads mounted on them. He knows of suppliers which will supply him with the actual fluorescent beads but he wants them already mounted. He thought a company called "Phoenix" may be able to help him but he can't locate the company. The slides (with fluorescent beads attached) will be used to assist in optimising the performance of a Bio-Rad Confocal MRC1000. Any help locating these slides would be appreciated.
Thanks in advance,
Sarah Ellis
Trescowthick Research Centre Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne 8006 Victoria Australia
Ed Sharpe asked about micromanipulators for light microscopes. Energy Beam Sciences distributes two lines of micromanipulation equipment. Information is available on-line at our web site (address in my .sig, below). Please contact me back-channel for additional information.
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
Does anyone have any suggested sources, recommendations, or comments regarding any software for tracking "cells" / motion analysis from microscopic images? Either live time, from video tape or a series of digital stills? Rather than trying to write software from scratch or as a series of macros it would be nice to find a ready to run package.
Aparently there used to be a package called "CellTrak" but a variety of web searches has resulted in nothing.
(Vendors should feel free to contact me as well.)
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
My first thought would be contaminated gas, either from a vacuum leak or a "bad" bottle of argon. The leak is more likely. With no Ar flow is your blank-off pressure the same low value as when working properly? I would also inspect the target/chamber for contamination of some sort. Another check.. If the pressure gauge is "off" you should notice a different current for the same voltage setting while sputtering.
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Hello to the coating experts:-
We have a Polaron diode sputter coater we use exclusively for gold coating for conventional 20 kV tungsten emitter SEM.
Lately the coatings being applied seem comparatively thin - instead of a shiny metallic gold coloured coat we get a semitransparent dark gray coat.
But we are coating using the same parameters as ever : 50 mA -at- 1.4 kV with 0.2 torr of argon.
This occurs even with plain glass specimens which should not be contributing any unwanted vapours.
I have checked out the pumping system and the argon gas bottle is labelled "high purity argon" and is quite new.
I would welcome suggestions as to the source of our problem.
Mel Dickson
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
id AA49470; Thu, 27 Aug 1998 08:08:05 -0400 Message-Id: {9808271208.AA49470-at-rose.muohio.edu} Received: from CASSERVER1/SpoolDir by casmail.muohio.edu (Mercury 1.32); 27 Aug 98 08:08:06 -5 Received: from SpoolDir by CASSERVER1 (Mercury 1.32); 27 Aug 98 08:07:43 -5 To: microscopy-at-Sparc5.Microscopy.Com, CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU
Looking for recomendations for a film scanner. The Leaf 45 is no longer in production. So far I have only found the Nikon LS-4500AF and the Polaroid Spirit Scan 45 Pro. Any others? Any recommendations?
Flatbeds are o.k. for 4X5's (we already have one) but would also like to be able to handle 35mm's as well (8x10 } 9x14 flatbed scanners just don't have the resolution for 35mm's).
Thanks again.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
Dear all, we found paracrystalline ER-structures in cells of the nectarie= s of plant flowers. Has anyone seen similar structures or knows about such things documented in t= he literature? Thanks for help Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=E4tsstrasse 25 Germany 33615 Bielefeld phone: 0521 1065592 fax: 0521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
} I would like to know a way to grind and polish diamond samples for a SEM } study. What do you want to do for diamond samples. If you just observe the morphology of the samples by SEM, the samples needn't polish and grind. I ever used an agate motar to grind diamond, but the agate motar was worn out.
You may increase the number of a particular virus if the grid has been treated with antibody, but, it does not stop TMV from attaching to the grid. Why not treat the extract with TMV antibody and centrifuge it. You should be able to get rid of all TMV if correct amount of antibody is used.
Ann Fook Yang EM Unit Eastern Cereal and Oilseed Research Centre Agriculture and Agri-Food Canada 960 Carling Ave Central Experimental Farm Ottawa, Ontario Canada K1A 0C6
} } } Debby Sherman {sherman-at-btny.purdue.edu} 08/26/98 05:49pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a user who wants to try to capture a specific virus from a prep contaminated with TMV. I believe there are methods involving placing a layer of antibody against a coat protein from the virus you want to capture on a grid. Then you float the grid on a droplet of the virus prep, wash off any unbound particles and negative stain. Hopefully this would leave only the desired virus on the grid. Does anyone use this technique and/or have a more detailed procedure?
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Maybe I'm going blind. I've been digging through my Noran Voyager manuals and I can't seem to find KeV and take-off angle limits for the Proza quant. routine. Does anyone know the limits or can they direct me to the correct page(s)?
Thanks,
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
I have a user who wants to try to capture a specific virus from a prep contaminated with TMV. I believe there are methods involving placing a layer of antibody against a coat protein from the virus you want to = capture on a grid. Then you float the grid on a droplet of the virus prep, wash off any unbound particles and negative stain. Hopefully this would leave only the desired virus on the grid. Does anyone use this technique and/or have a more detailed procedure?
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu=20 Purdue University =20 1057 Whistler Building West Lafayette, IN 47907-1057
The technique you descirbe is the "immunosorbent technique" and relies as you know on the fact that proteins tend to stick to the coated grids, especially a plastic such as formvar (which can be carbon coated to make it more stable- but then proteins do not stick as well). Antibodies appear to stick well to formvar-carbon and in my experience, once they are "on" its pretty well impossible to get them off!
This method will not completely get rid of your TMV contamination, that will stick to the grid as well (but will at least be a useful "standard" for magnification calibration!), however, the immunosorbent technique should enrich the number of specifically adsorbed particles which react with the antiserum. This is great for negative staining viruses when you only have a small volume of very dilute particles- so long as you have an antibody which you know reacts with the surface of the particle.
You will have to play with the conditions to get the optimum results with your anitbody, try a few different dilutions of it.
To start with, I suggest the following:
1. Float the grids on your antiserum- 10 min(all reactions carried out in humid chamber to prevent drying out) 2. Wash in PBS containing .1 per cent BSA 30 seconds 3. Float grids (antiserum coated side down!) on your virus preparation, for 5 minutes. 4. Blot of excess, float on PBS/BSA as above to wash. 5 Brief wash in deionised warter (optional) 5. Negative stain using your favourite stain UA, PTA etc.
best of luck!
Dr Timothy F. Booth Canadian Food Inspection Agency National Centre For Foreign Animal Disease Suite T2300 1015 Arlington St. Winnipeg Manitoba R3E 3M4 CANADA http://www.hc-sc.gc.ca/main/lcdc/web/bmb/fedlab_e.html#toc email tbooth-at-em.agr.ca Tel 204 789 2022 (office) Tel 204 789 2038 (lab) Fax 204 789 2038
You might want to take a look at the UMAX Powerlook 3000. Paper specs look pretty good and at $6995 (list), the price is less than the Polaroid.
http://www.umax.com/
Henk
At 08:07 AM 8/27/98 -0500, you wrote: {snip} } } Looking for recomendations for a film scanner. The Leaf 45 is no longer in } production. So far I have only found the Nikon LS-4500AF and the Polaroid Spirit Scan } 45 Pro. Any others? Any recommendations? } {snip} } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } } "640K ought to be enough for anybody." } -- Bill Gates, 1981 } Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
Another flatbed scanner which we considered a few years ago:
Dicomed 6300T: 35 mm to 4"x5", 12 bits/pixel, 6000 pixel CCD optical resolution (landscape mode ): 4233 dpi for 35 mm; 1270 dpi for 4"x5" Dicomed, 12270 Nicollet Ave., Burnsville, MN 55337, (612) 895-3000
} ------------------------------------------------- } Looking for recomendations for a film scanner. The Leaf 45 is no } longer in } production. So far I have only found the Nikon LS-4500AF and the Polaroid } Spirit Scan } 45 Pro. Any others? Any recommendations? } } Richard E. Edelmann, Ph.D. -------------------------------------------------
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello to the coating experts:- } } We have a Polaron diode sputter coater we use exclusively for gold coating } for conventional 20 kV tungsten emitter SEM. } } Lately the coatings being applied seem comparatively thin - instead of a } shiny metallic gold coloured coat we get a semitransparent dark gray coat. } } But we are coating using the same parameters as ever : 50 mA -at- 1.4 kV with } 0.2 torr of argon. } } This occurs even with plain glass specimens which should not be } contributing any unwanted vapours. } } I have checked out the pumping system and the argon gas bottle is labelled } "high purity argon" and is quite new. } } I would welcome suggestions as to the source of our problem. } } Mel Dickson } } } ***************************************************** } Mel Dickson, } Director. } Electron Microscope Unit, } University of New South Wales. } Sydney NSW 2052 Australia } } Phone (+612) 9385-6383 } Fax (+612) 9385-6400 } Website {http://srv.emunit.unsw.edu.au} } ***************************************************** } } Hi Mel,
Check for an air leak on your Argon input line particularly between the chamber and the leak valve. Black gold is always caused by air leaks on our sputter coater.
Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
thanks for the info! I am trying to do things on a budget and am willing to accept old crusty equipment that even needs work done to it! I wish I had the budget for new equipment but sometimes we make do as best we can!
thanks though! Dear fellow microscopists,
Ed Sharpe asked about micromanipulators for light microscopes. Energy Beam Sciences distributes two lines of micromanipulation equipment. Information is available on-line at our web site (address in my .sig, below). Please contact me back-channel for additional information.
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com *
Dear Mel, If the operating parameters are the same, then the obvious culprits are the gold target getting thin or gone, or contamination of the gold target by vapours of some kind. Try cleaning the target with acetone or clean alcohol. You can even polish it lightly. You might also check all the high voltage contacts and the contact of the gold to the plate. Good luck. } } } Hello to the coating experts:- } } We have a Polaron diode sputter coater we use exclusively for gold coating } for conventional 20 kV tungsten emitter SEM. } } Lately the coatings being applied seem comparatively thin - instead of a } shiny metallic gold coloured coat we get a semitransparent dark gray coat. } } But we are coating using the same parameters as ever : 50 mA -at- 1.4 kV with } 0.2 torr of argon. } } This occurs even with plain glass specimens which should not be } contributing any unwanted vapours. } } I have checked out the pumping system and the argon gas bottle is labelled } "high purity argon" and is quite new. } } I would welcome suggestions as to the source of our problem. } } Mel Dickson } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Is there any chance that the Au target has worn through in spots? If so, it may be possible that sputtering is taking place with the underlying metal instead of or in addition to the Au.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML--Biology New Mexico State University Las Cruces, NM 88003
Flatbed scanners with transparency capability may offer the capability you need at much lower price than a 4 x 5 film scanner or drum scanner. Several models now costing a few $K have optical resolutions to } 2K and 14-bit grayscale. For a recent review, see Macworld October, 1998, pp77-81. If possible, try before you buy and look carefully at the control software. Scanning times can be longer than you can tolerate (How long does it actually take to scan each negative?), and manual scanner controls tend to be primitive or arcane (often both). Even 12-bit grayscale is usually sufficient for high-contrast negatives including TEM diffraction patterns if these are properly exposed. Manual exposure control would be useful for dealing with over/under-exposed film, but is seldom included.
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA
---------- From: Hendrik O. Colijn Sent: Thursday, August 27, 1998 4:23 PM To: edelmare-at-muohio.edu Cc: microscopy-at-Sparc5.Microscopy.Com Subject: Re: 4" x 5" film scanners?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Richard,
You might want to take a look at the UMAX Powerlook 3000. Paper specs look pretty good and at $6995 (list), the price is less than the Polaroid.
http://www.umax.com/
Henk
At 08:07 AM 8/27/98 -0500, you wrote: {snip} } } Looking for recomendations for a film scanner. The Leaf 45 is no longer in } production. So far I have only found the Nikon LS-4500AF and the Polaroid Spirit Scan } 45 Pro. Any others? Any recommendations? } {snip} } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } } "640K ought to be enough for anybody." } -- Bill Gates, 1981 } Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
One way is to apply the diamond surface to a hot iron alloy flat - this has been done at the naval research lab and I think a guy at auburn has a patent on the process. Ther may be some articles in the literature on this.
Milo
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The most common is etchant nital - 2% nitric acid in methanol. swab the polished surface with the etchant for 2 - 10 seconds. see the metals handbook. the carbide will always be evident, but in addition the ferrite grain boundaries will be evident.
Milo
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I represent Micro Star Diamond Knives, and Technical Manufacturing Corporation and their "MICROg" Vibration Isolation systems. If I can be of any assistance to any one please contact me {tcooper-at-sprintmail.com}
I use a Polaroid SprintScan and am fairly happy with it. Here are a couple of high end scanners that I had gotten info on, but cannot find anymore.
Scitex Smart Scanner: this is a flat bed. Visit Scitex's web site. http://www.scitex.com Ask for info and they will send a demo CD about the scanner. I believe that you are restricted to using a Mac system with this.
Scanview Scanmate 5000 or Scanmate 3000: this is a drum scanner. Visit this site http://www.medgraphix.com/scanmate3000.htm for info on the 3000. It is about $20,000.
I did a search on the web for the above two scanners and found the following sites for vendors of scanners that have a variety of scanners for sale.
"The Scanner Guys, 508-456-9220": http://www.ultranet.com/~bgriffin/ They have a whole bunch of different scanners, but there isn't much info on their page about each one. They want you to call.
Here is another that has new and used scanners of all types including used LeafScan 45's. http://www.promarketinc.com You should be able to find something in your price range with one of these places.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: "edelmare-at-casmail.muohio.edu"-at-sparc5.microscopy.com To: microscopy-at-sparc5.microscopy.com; CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU
Looking for recomendations for a film scanner. The Leaf 45 is no longer in production. So far I have only found the Nikon LS-4500AF and the Polaroid Spirit Scan 45 Pro. Any others? Any recommendations?
Flatbeds are o.k. for 4X5's (we already have one) but would also like to be able to handle 35mm's as well (8x10 } 9x14 flatbed scanners just don't have the resolution for 35mm's).
Thanks again.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
I say make your own! Get the beads (I use Bangs Labs, here in the USA somewhere), dilute the suspension at least 100:1, or even more, drop on slide, let dry, add epoxy and cover glass, let set.....confocal. Used it to evaluate confocals, bought a BioRad ourselves. We are doing volume measurements, so calibrated beads with known std. dev. were useful. We even deformed the beads (heat & pressure) to test ability to find volumes of irregular shapes. Don't mount in oil or water, as the cover glass sticks to lens imersion oil and the little balls scoot all over when moving the stage. Dave Pevear, Houston, Texas
Although this is not a specific microscopy question, somebody of you might have an answer. We just started to study apoptosis in alveolar epithelial cells. We are looking for a reliable method to induce apoptosis in our cell culture system (L132). Has anyone any experience about how to reliably induce apoptosis in the human embryonic lung epithelial cell line L132 ? Any information will be greatly appreciated.
Thank you very much in advance.
Heinz
*********************************************************************** Dr. Heinz Fehrenbach Institute of Pathology University Clinics "Carl Gustav Carus" Technical University of Dresden
We have a Siemens Elmiskop 102 electron microscope. We use pointed filaments. We distinguish two types of filament : * A one-piece filament where the tip is mechanically thinned * A two-pieces filament where the tip is welded on the loop.
The one-piece filament is the only one which is suitable for our microscope.
Does anyone have any information about this type of filament ?
Thanks in advance for your answers
Sincerelly yours
Philippe Drouillon Solvay Research and Technology Rue de Ransbeek, 310 B-1120 Brussels (Belgium) tel : (0032)2.264.24.47 mailto:philippe.drouillon-at-solvay.com
I am curious to know how does one define Negroid, Caucasian and Mongoloid SCIENTIFICALLY? This smacks of Prof. Rushton's preposterous theories about testosterone and IQ of "races". And statistically, the differences within a defined group is not different ENOUGH to the differences between. So how does one do it? ...by intuition?
Azriel wrote:
My appologies for the lateness in this reply, I have been out of the country. As an old and still practicing classical hair comparison person my views may be seen by many as biased.
The simple answer is that there has been found a relationship between cross sectional shape and racial characteristics. Flat being associated with the Negroid race, oval with the Caucasion race, and round with Mongoloid.
To quote from "MICROSCOPY OF HAIR, A Practical Guide and Manual" put out by the FBI:
"A human hair may be classified according to its racial characteristics as being Caucasian, Negrod or Mongoloid origin. In some instances, the racial characteristics exhibitied by the hair specimine may not be clearly defined indicting the sounce of the particular hair may be of mixed racial origin."
and
"Again, it is pointed out thet even when racial characteristics are not clearly defined, it is significant when these characteristics are consistent between the hair in questiion and the hair of known origin."
Okay, probably more than you wanted to know. Good luck.
Shalom from Jerusalem, Azriel +++++++++++++++++++++++++++++++++++++++++++++++++++++ Major Azriel Gorski, Head Fibers and Polymers Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem, ISRAEL
azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739
What lies behind us and what lies before us are tiny matters compared to what lies within us. Ralph Waldo Emerson ++++++++++++++++++++++++++++++++++++++++++++++++++++
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
You mention that the cylinder of argon is new, it seems possible and likely that this is coincident with the problem and it may be that there is a high water content in the argon. Most labs will have a nitrogen cylinder available and as a check this could be used to see if the problem remains.
Other possibilities are the rotary pump backstreaming oil into the plasma if left to run at ultimate pressure, or an 'O' ring breaking down within the target area.
The operating parameters of 1.4Kv at 50mA would indicate a very old E5000 coater and it is possible that there are vacuum leaks around the target when hot. A good service with 'O' ring replacement may also cure the problem.
Concerning the possibility of non-target material being sputtered through holes in the target, it can occur with some designs of target and target holders. For example another previous Polaron model, the SC500, used a target material bonded to a holding plate using a silver loaded epoxy resin. With this design it was possible to sputter silver and resin components (i.e. contaminantsl) through holes eroded in the target. This is unlikely with the E5000, or with similar sputter coaters that do not use silver bonding material. Although it would seem possible that material from the target holder (i.e.aluminium) could be sputtered through holes in a worn or damaged target, the power (e.g. 1.4Kv at 50mA) and design of such coaters would normally not preclude this.
Best regards M.J.Wombwell Polaron range Business Manager http://www.vgmicrotech.com/polaron-range
I will not get into the anthropological "p'n contest" that has gone on in the recent past on "defining race." As we both know from South Africa and from the experiences of trying to define, scientifically measure, and deal with races in the 1930's/40's and others ... the term and concept of race can be sensitive and has been, to be mild, badly misused.
I will only state that there are accepted and usable definitions, in the common use, if not the scientific. Those definitions have characteristics associated with them.
I feel as a person and even a a scientist, I can be conversant wi, recognize and use those concepts.
Shalom from Jerusalem, Azriel
On Fri, 28 Aug 1998, Quirina Roode wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Azriel Gorski, } } I am curious to know how does one define Negroid, Caucasian and } Mongoloid SCIENTIFICALLY? This smacks of Prof. Rushton's preposterous } theories about testosterone and IQ of "races". And statistically, the } differences within a defined group is not different ENOUGH to the } differences between. So how does one do it? ...by intuition? } } Azriel wrote: } } My appologies for the lateness in this reply, I have been out of the } country. As an old and still practicing classical hair comparison person my } views may be seen by many as biased. } } The simple answer is that there has been found a relationship between cross } sectional shape and racial characteristics. Flat being associated with the } Negroid race, oval with the Caucasion race, and round with Mongoloid. } } To quote from "MICROSCOPY OF HAIR, A Practical Guide and Manual" put out by } the FBI: } } "A human hair may be classified according to its racial characteristics as } being Caucasian, Negrod or Mongoloid origin. In some instances, the racial } characteristics exhibitied by the hair specimine may not be clearly defined } indicting the sounce of the particular hair may be of mixed racial origin." } } and } } "Again, it is pointed out thet even when racial characteristics are not } clearly defined, it is significant when these characteristics are } consistent between the hair in questiion and the hair of known origin." } } Okay, probably more than you wanted to know. Good luck. } } } Shalom from Jerusalem, } Azriel } +++++++++++++++++++++++++++++++++++++++++++++++++++++ } Major Azriel Gorski, Head } Fibers and Polymers Laboratory } Division of Identification and Forensic Science } Israel National Police } Jerusalem, ISRAEL } } azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739 } } What lies behind us and what lies before us are tiny } matters compared to what lies within us. } Ralph Waldo Emerson } ++++++++++++++++++++++++++++++++++++++++++++++++++++ } } } } \|||/ } (o o) } *----oOO------(_)-OOo---------* } } Quirina I. Roode } PhD student: Cement Chemistry } *-----------------------------* } Department of Civil Engineering } University of the Witwatersrand } P O Wits, 2050, Johannesburg } SOUTH AFRICA } *-----------------------------*Oooo. } ROODE-at-civen.civil.wits.ac.za ( ) } Tel: +27-(0)11-716-2478 (w) ) / } Fax: +27-(0)11-339-1762 (w) (_/ } *.oooO------------------------* } ( ) } \ ( } \_) }
I am going to put my full 2D code for Direct methods (surfaces and TED data) out as Public Domain code soon. I will have a used interface similar to Shelx, and output either of semper images (Fortran files that can be read by other programs if someone writes an interface), "miff" files for ImageMagick or hkl,F,phase.
Any volunteers for testing it? The code is Fortran, pretty much platform independant, but parts of it are specificaly UNIX based.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Philippe Drouillon asked: } We have a Siemens Elmiskop 102 electron microscope. We use pointed } filaments. } We distinguish two types of filament : } * A one-piece filament where the tip is mechanically thinned } * A two-pieces filament where the tip is welded on the loop. } } The one-piece filament is the only one which is suitable for our } microscope. } } Does anyone have any information about this type of filament ?
Energy Beam Sciences offers a proprietary "SG" filament loop, which fits this description. The tip of the filament loop is etched into a "spade" shape. Higher brightness can easily be acheived with a filament of this type.
Please contact me back-channel for additional information.
I have been following this thread with interest. I suspect that by now the sixth grader is wondering how, what he meant as a simple question (I don't think he was thinking of submitting this for publication), has turned into a racially spiked issue about testosterone and IQ. My guess is that by now the student is asking himself a deeper question : "Are microscopists for real ???"
My two cents on the issue.
Jordi Marti
---------- } From: Quirina Roode To: microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
Does anyone have any experience with converting an older SEM so that pc-based images can be collected? Two questions: !) What issues are important? 2) Any recommendations on companies that sell the conversions?
Yes, if at all possible, you should have a biohazard hood for dealing with unfixed samples, especially lung samples that may contain such things as TB spores that are easily aerosolized. All other specimens can be moved to a fume hood once they're fixed in glut (except for brain samples which may have spongioform encephalopythy. e.g., Creutzfeld-Jacob disease).
My Surgical Pathology EM Lab processes 300-400 tissue samples per year. All come in glut. They have no biohazard hood. My EM Diagnostic Virology Lab processes around 800 nonfixed specimens per year; all samples go into the biohazard hood where negative stains are processed and tissue samples are placed into glut. Fixed specimens are transferred to the fume hood for further processing.
Sara Miller (Director, Surg Path EM Lab & EM Diag Virol Lab) address below
On Mon, 11 May 1998, Richard Easingwood wrote:
} Date: Mon, 11 May 1998 14:41:57 +1200 } From: Richard Easingwood {richard.easingwood-at-stonebow.otago.ac.nz} } To: MICROSCOPY-at-sparc5.microscopy.com } Subject: Biohazard cabinets for EM labs } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello microscopists, } } We have been given the go ahead to upgrade our electron microscope } preparation laboratory area including the choice of installing new } fumehoods and/or biohazard cabinets. } } At present we have two fume hoods and no biohazard cabinets. We are } thinking of asking for four fume hoods or, alternatively, three fume hoods } and one biohazard cabinet. Our feelings were that the biohazard cabinet } might be safer considering the human biopsy material we deal with, but } maybe it would be no safer than a good fume hood. } } My question is: should we consider a biohazard cabinet in place of a } fumehood given that many of the biohazards dealt with in tha lab are also } in toxic fixatives or solvents? Do other EM labs use biohazard cabinets in } preference to fume hoods? } } (By fume hoods I mean that the fumes are extracted from the room and } released outside whereas a biohazard cabinet filters the air and returns it } to the room). } } Many thanks for any thoughts you might have on this matter. } } Richard } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences } University of Otago } PO Box 913, Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } e-mail: richard.easingwood-at-stonebow.otago.ac.nz } } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I would like to think it would be a simple matter any more. We had a JEOL U3 set up for digital control back in 1981. It was relatively straightforward to find the connections for scan control and to have the scanning hardware set up to work the same voltage range. Unfortunately the computers were not so friendly then, nor were the electronics all that fast. Present D/A cards should be able to do much of that all from within the PC box.
The biggest issues would probably be software, speed and allowance for hysteresis in the scan coils. Taking those in reverse order, I know even our JEOL 840 can be significantly off at high scan speeds, but normal active digital scanning systems will probably not be near that fast. There can be a significant amount of overhead in stepping from point to point so that scans can be slow for 1024 pixels across the image. We find 100 us of dwell per point is plenty from a signal standpoint. I don't know how fast the scanning can reasonably be performed in conjuction with image digitization.
Software is a non-trivial issue. Being a tinkerer, I could probably come up with a hardware solution in a reasonable amount of time. However, the software will probably determine the overall satisfaction with such a system. It is probably the greater investment of energy.
Therefore, IMO, you should probably evaluate commercially available systems to find one with software that meet your criteria (usability, database functions, etc.), and then pursue the question of matching the hardware up to your microscope.
FWIW, we are happily using the Quartz PCI passive imaging system. Rather than taking over control of the beam, the system passively monitors the scan and records the image. I would suppose there is enough latitude in their hardware adjustments to match the raster of virtually any microscope with minimal bother. (Y'all can insert the standard disclaimer here.)
At 12:06 PM 8/28/98 -0500, you wrote: } ------------------------------------------------------------------------ } Does anyone have any experience with converting an older SEM so that } pc-based images can be collected? Two questions: !) What issues are } important? 2) Any recommendations on companies that sell the conversions? } } Thanks, } } Robin Griffin
We have had excellent success with the 4pi System's Spectral Engines for retrofitting old systems as well as new installations. It is a _VERY_ cost competitive option, particularly in terms of flexibility and capabilities. If you already have a scan control interface, hooking up a Spectral Engine is very straightforward. They also have a lot of experience at getting into the depths of older instruments.
http://www.4pi.com 919-489-1757
Bill Heeschen The Dow Chemical Company
} I would like to think it would be a simple matter any more. We had a } JEOL U3 } set up for digital control back in 1981. It was relatively } straightforward } to find the connections for scan control and to have the scanning } hardware } set up to work the same voltage range. Unfortunately the computers } were not } so friendly then, nor were the electronics all that fast. Present D/A } cards } should be able to do much of that all from within the PC box. } } The biggest issues would probably be software, speed and allowance for } hysteresis in the scan coils. Taking those in reverse order, I know } even our } JEOL 840 can be significantly off at high scan speeds, but normal } active } digital scanning systems will probably not be near that fast. There } can be a } significant amount of overhead in stepping from point to point so that } scans } can be slow for 1024 pixels across the image. We find 100 us of dwell } per } point is plenty from a signal standpoint. I don't know how fast the } scanning } can reasonably be performed in conjuction with image digitization. } } Software is a non-trivial issue. Being a tinkerer, I could probably } come up } with a hardware solution in a reasonable amount of time. However, the } software will probably determine the overall satisfaction with such a } system. It is probably the greater investment of energy. } } Therefore, IMO, you should probably evaluate commercially available } systems } to find one with software that meet your criteria (usability, database } functions, etc.), and then pursue the question of matching the } hardware up } to your microscope. } } FWIW, we are happily using the Quartz PCI passive imaging system. } Rather } than taking over control of the beam, the system passively monitors } the scan } and records the image. I would suppose there is enough latitude in } their } hardware adjustments to match the raster of virtually any microscope } with } minimal bother. (Y'all can insert the standard disclaimer here.) } } At 12:06 PM 8/28/98 -0500, you wrote: } } --------------------------------------------------------------------- } --- } } Does anyone have any experience with converting an older SEM so that } } pc-based images can be collected? Two questions: !) What issues } are } } important? 2) Any recommendations on companies that sell the } conversions? } } } } Thanks, } } } } Robin Griffin } }
Cheers, Jordi! Let's get back on topic folks. There may be a place for the discussion that followed posting by the 6th grade student but this list server isn't it.
********************************************************************** Rhonda L. Callender Inorganic & Materials Chemistry Rice University 6100 Main Street MS60 Houston, TX 77005
On Fri, 28 Aug 1998, Marti, Jordi wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have been following this thread with interest. I suspect that by } now the sixth grader is wondering how, what he meant as a simple } question (I don't think he was thinking of submitting this for } publication), has turned into a racially spiked issue about } testosterone and IQ. My guess is that by now the student is asking } himself a deeper question : "Are microscopists for real ???" } } My two cents on the issue. } } Jordi Marti } } } ---------- } } From: Quirina Roode } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Shape of Hair Question from a 6th Grade Student } Date: Friday, August 28, 1998 7:03AM } } ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- }
We converted an 1981 ISI DS-130 to a digital acquisition system that we are very pleased with. It has been in use for about 5-6 years. The system (Quartz PCI passive acquisition system) was developed in Canada and is available from:
At 12:06 PM 8/28/98 -0500, rgriffin-at-eng.uab.edu"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We converted a 1981 ISI DS-130 to a digital acquisition system that we are very pleased with. It has been in use for about 5-6 years. The system (Quartz PCI passive acquisition system) was developed in Canada and is available from:
At 12:06 PM 8/28/98 -0500, rgriffin-at-eng.uab.edu"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If a lab has an SEM or light microscope (up to four), SEMICAPS Image Capture Software will archive your image, paste the image to a report, and use keyword searches for future image retrieval. Depending on your network (internet or intranet) system, SEMICAPS can send the image as a TIF or JPEG file.
SEMICAPS consists of three systems:
The passive system will tap into the video signal of the SEM monitor.
The active system controls the electron beam and is most useful for background noise removal.
The color-capture system will connect up to four light microscopes and includes a "snap feature" that allows each user independent control of the image capture process.
If your image system requires: archival, noise reduction, image processing, measurement, annotation, and printout, SEMICAPS is the system most used.
Give Bruce, Eric or Elie a call if you would like a no-cost demo. 408-986-0121
I am new to the list and to microscopy (just got a used scope for home use). Can anyone help me find an illuminator for an AO student scope of ca. 1950 vintage? This is a black monocular scope with substage condenser. Do I need something particularly for this model or would other illuminators work? If so, how can I tell before ordering? Thanks for your indulgence.
Hi there. Can you email me a pic of the scope? They made several during that time. Please send Pic to couryhouse-at-azonline.com I will check my old catalogs and see if I have something in the Parts box here that might be made to work. thanks Ed Sharpe
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings! You don't say whether your scope (which will give you a lot of enjoyment) has a mirror, or was originally fitted with a substage illuminator. If you have a mirror, you can start with almost anything - even a gooseneck lamp. You can get something more sophisticated when you've developed your own needs and opinions. You do need a book or two. I suggest that you look at the Project MICRO bibliography (address below). There are several good books for adult hobbiests listed; you might start with Nachtigall (Section IIA).
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Discussions at MSA in Atlanta identified a void in high resolution "in vitro" microscopy.
Recent progress made in bio in vitro AFM are meeting this need. Some applications presented clearly point to new microscopy capabilities. For example: 1- time lapsed in vitro imaging of katanin disintegration of microtubule (Vail group UCSF) 2- molecular recognition force microscopy which characterized antibody - antigen binding force in vitro (Schindler group Linz Austria) 3- protein folding force characterization (Trinick group Leeds UK)
Recent innovations in AFM are 1- magnetic AC mode and 2- optimizing the microscope for fluid operation a) under environmental, b) temperature control, and c) electrical properties characterization (elecrophysiology)
These innovations have filled a void in high resolution "in vitro" microscopy.
Please contact me via e-mail or at (800) 819-2519.
This is just a short note to ask your permission to send you details about our latest skywatch gathering for West Central Indiana.
If you are interested and/or would like to attend, reply to this message. We'll send you date, time and location information. If you are not interested, do nothing and you will not receive the information.
Also, if you reply, rest assured that your e-mail address will not be broadcast, sold or otherwise distributed in any manner to any e-mail "SPAM" list. Your reply will return to a real person that will read it and send you the information, if that is what you wish.
I suppose I have to take some of the responsibility of this off- topic racial discussion. Couldn't help it...it is so facinating...and just maybe it is not so off topic, because the issue is whether we can answer these questions under the microscope or can we not. Science isn't all about what we can see and what we can't, it isn't just about data, it's also about interpretation or even inferrence. My apologies...
Cheers, Quirina
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
In our research we study particles under an ordinary optical microscope at 20 times magnification. These particles are placed in a so-called micro-reactor that is closed with a glass lid to be able to see the particles. The glass is rather thick (8 mm!) because the reactor is pressurized (max. pressure 30 bar). The problem is that around the particles a 'dispersed cloud' of white light is observed. This light makes it impossible for us to observe the particles in detail.
We are not sure what causes this phenomenon. We think it may have two reasons. First, it may be caused by the diffraction of the light on the glass/air surface, for the glass disturbs the direction of the light. Because of this not all light emitted by a point light source will converge to one point. We didn=92t succeed in deriving the theoretical point spread function. However, we could derive that a point light source was spread out over a large distance, but it also could be derived that most of the light can be focussed to the middle of the =91spot=92 in an infinite intensity top. Second, it may be possible that the glass lid is made of (relatively) low quality glass. It might not be adequately polished, or may contain impurities. We doubt whether this is the reason, however we already ordered a higher quality glass.
Does anybody have another idea what may cause this problem, or even knows it for sure? More important, does anybody know a solution, is it possible to correct for this? Is there anybody else who uses thick cover slips (owing to high/low pressures)? I believe there are lenses that correct for cover slip thicknesses of 1.5 mm, is this because of the same reason? An obvious solution may be a thinner cover slip. Does anybody know an extremely strong material?
Some details about the used microscope: Zeiss AxioTech Vario, 20x LD Epiplan lens (number 442840, N.A =3D 0.4, WD =3D 9.8 mm).
We hope someone can help us with this problem!
Thanks in advance for any help or suggestions you may have.
Ronald Capel
University of Twente Faculty of Chemical Technology PO-Box 217, 7500 AE, Enschede, Netherlands r.capel-at-student.utwente.nl
I have been using the Printerface program, purchased from GW Electronics,6981 Peachtree Industrial Blvd., Norcross, GA 30092-3601. Phone is (800)325-5556, Fax is (770)449-0284. This is a passive system (i.e., it does not take control of microscope operation), and the image can be stored or sent in several file formats. The price was reasonable, and my customers have been pleased with the system.
Leslie
Leslie Eibest Zoology Dept., Box 90325 Duke University Durham, NC 27708 USA (919) 684-2547 leibest-at-duke.edu
I assume you are referring to the ER itself being paracrystalline, and not to crystals of protein or other substances that can occur within the lumen of the ER.
Here are three references that may be of interest (their reference lists will guide you to others):
Anderson RGW, Orci L, Brown MS, Garcia-Segura LM, Goldstein JL, 1983. Ultrastructural analysis of crystalloid endoplasmic reticulum in UT-1 cells and its disappearance in response to cholesterol. J Cell Sci 63:1-20.
Kochevar DT, Anderson ROW, 1987. Purified crystalloid endoplasmic reticulum from UT-1 cells contains multiple proteins in addition to 3-hydroxy-3-methylglutaryl coenzyme A reductase. J Biol Chem 262:10321-10326.
Fringes B, Gorgas K, 1993. Crystalloid smooth endoplasmic reticulum in the quail uropygial gland. Ann Anatomy 175:231-235.
Regards, Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
} Dear all, we found paracrystalline ER-structures in cells of the } nectaries of plant flowers. Has janyone seen similar structures or knows } about such things documented in the literature? Thanks for help
} Bernward Laube } University of Bielefeld } Faculty of Biology } Department Plant Morphology and Cell Ultrastructure } Universit=E4tsstrasse 25 } Germany 33615 Bielefeld } phone: 0521 1065592 } fax: 0521 1066039 } e-mail: b.laube-at-biologie.uni-bielefeld.de } http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws } =20
We are looking for sands from around the world to support the GEMS/Proj= ect Micro elementary science program, MICROSCOPIC EXPLORATIONS. If anyone = would like to collect a handful of sand from Cancun for this project it would= be greatly appreciated. If you can send some sand please contact me first= so I can avoid receiving to many samples. The samples can be sent to: =
Dear All: We need to prepare TEM specimen to study a thin film on single crystalline graphite. One of the methods to prepare a sample is to cleave graphite using adhesive tape. Does anybody know the tape which leaves no glue after dissolving? Thanks, Serge.
I did this another way on single crystal MoS2 using low temperature wax and Post-it notes. The MoS2 cleaves like graphite.
I took the MoS2 and waxed it down to a glass microscope slide using low temperature melting wax (glycol phthalate) from SouthBay Technology. After it cooled, I just repeatedly stripped off layers with new areas of Post-it notes until what remained was optically transparent. I dissolved the wax in acetone. I can't remember if I just dipped out the films with a grid or collected them using a loop. I think that I tried both. You could use a variation if you lowered the acetone bath level so that the films were collected on a grid support plate.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Serge Oktyabrsky To: Microscopy-at-sparc5.microscopy.com
Good day all,
Here's a question for the TEM/STEM crowd (in my case a Philips CM30).
I'm looking for advice on what is apparently an alignment problem that results in the shift of my PEELS signal, up and down on the energy scale (by about 1.5-2 eV) as the STEM beam is rastered across a line. Up 'till now, I thought that I had figured out how to do a decent STEM alignment (decent resolution in STEM images up to about 400kX) but apparently something is still off. I have tried numerous realignments of the pivot points and rotation centering- along with the DESCAN set up- with little luck. Does anyone have any advice before I have to start thinking about what I'm doing?
Specifics; Philips CM30 (300kV) Gatan 666 spectrometer. Following STEM alignment procedures given from Philips (if anyone wants details, I can write them out and I'd be happy to discuss them since the problem could be a result of interpretation). Using DESCAN mode as described in CM30 instruction manual for STEM-PEELS. -checking the voltage rotation centering and beam pivot points... is there a problem going from doing this on the view screen which is at a different height from the spectrometer entrance aperture?
I've been at this for a day now, and while I still think I can figure it out, I'd be extremely happy to get some advice if it is easy of obvious to anyone with more STEM-PEELS experience. I'd be happy to discuss via email, or if anyone wants to talk me through it, I'd be happy to return a page.
Brendan Foran, Ph.D. Materials Analysis Group SEMATECH Austin, TX
phone: 512-356-3936 (never there, but messages can be left on voicemail) pager: 1-888-731-4459 ________________________________________________________________________ ___ For fun: ...I can only be glad that I do not use the "CM30" described at: http://www.neatworld.com/incont.htm ..parallel universes?
[skip] } An obvious solution may be a thinner cover slip. Does anybody } know an extremely strong material? } Either fused quartz or, better yet, saphire are extremely strong. I don't know, however, what area one can use to contain the pressures you're working with. Good luck. Yours, Bill Tivol
With 120 film (2 and 1/4 inch square), what does the 120 refer to? It can't be 120 mm frame size. Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
A parallel plate of glass will introduce spherical aberration into a diverging beam. This is why high NA lenses are designed with a cover slip in mind if one is to be used. In your extreme case you will have roughly 200 microns of spherical aberration if your particles are in air, about half that if they are in liquid.
This is a tough problem. One solution would be to make a window from a concentric meniscus lens--where both surfaces have the same center of curvature. Then if the particles are at the center of curvature and the rays hit the window perpendicular to its surface you should eliminate the spherical aberration, or at least minimize it. This will make a much stronger window, although for your geometry it will be small.
If you have liquid in your chamber a plano-convex lens might suffice.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
My guess would be the image is suffering from sherical aberation. Perhaps you could find an objective off an inverted scope that has a correction collar that will allow you to adjust the ojective to the cover glass thickness. I doubt it can adjust to 8mm however.
bob Derm Imaging Center U of W
On Mon, 31 Aug 1998, Ronald Capel wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Dear all, } =20 } In our research we study particles under an ordinary optical } microscope at 20 times magnification. These particles are } placed in a so-called micro-reactor that is closed with a } glass lid to be able to see the particles. The glass is rather } thick (8 mm!) because the reactor is pressurized (max. } pressure 30 bar). The problem is that around the particles a } 'dispersed cloud' of white light is observed. This light makes } it impossible for us to observe the particles in detail. } =20 } We are not sure what causes this phenomenon. We think it may } have two reasons. } First, it may be caused by the diffraction of the light } on the glass/air surface, for the glass disturbs the direction } of the light. Because of this not all light emitted by a point } light source will converge to one point. We didn=92t succeed in } deriving the theoretical point spread function. However, we } could derive that a point light source was spread out over a } large distance, but it also could be derived that most of the } light can be focussed to the middle of the =91spot=92 in an } infinite intensity top. } Second, it may be possible that the glass lid is made of } (relatively) low quality glass. It might not be adequately } polished, or may contain impurities. We doubt whether this is } the reason, however we already ordered a higher quality glass. } =20 } Does anybody have another idea what may cause this problem, or } even knows it for sure? More important, does anybody know a } solution, is it possible to correct for this? Is there anybody } else who uses thick cover slips (owing to high/low pressures)? } I believe there are lenses that correct for cover slip } thicknesses of 1.5 mm, is this because of the same reason? An } obvious solution may be a thinner cover slip. Does anybody } know an extremely strong material? } =20 } Some details about the used microscope: Zeiss AxioTech Vario, } 20x LD Epiplan lens (number 442840, N.A =3D 0.4, WD =3D 9.8 mm). } =20 } We hope someone can help us with this problem! } =20 } Thanks in advance for any help or suggestions you may have. } =20 } Ronald Capel } =20 } University of Twente } Faculty of Chemical Technology } PO-Box 217, 7500 AE, Enschede, Netherlands } r.capel-at-student.utwente.nl } =20 } =20 } =20
In a message dated 98-08-31 17:50:29 EDT, bozzola-at-siu.edu writes:
{ { With 120 film (2 and 1/4 inch square), what does the 120 refer to? It can't be 120 mm frame size. Thanks.
This interesting little bit of confusion dates back to the commercialization of cameras and films, and we have Kodak to thank.
In the beginning (about 1898-1910) not all films fit all cameras, and the situation became worse as new cameras were introduced. Kodak decided to start numbering all of their films which were wound on flanged spools, beginning with the number 101, which was a standard spool of film that fit a specific camera. We still use this convention today. Several of the intermediate types have been dropped, and I think 127 film is scheduled to bite the dust in the year 2000.
You can learn more than you want about this bit of history by visiting:
Hi Folks, I have been trying to use EMbed-812 for TEM on C.elegans. I am using an ultracut E, with a glass knife and I just can not seem to get any thin sections using this resin.Poly/Bed 812 has worked fine, but is a little too viscous. I have tried both medium and hard mixtures to no avail. Does anyone have any ideas, I don't have access to a diamond knife.
THANK-YOU to all those who responded to my query on where to purchase slides with fluorescent beads on them for use in confocal calibration. I was asked by a few people to let others on the list know where to purchase these slides from. I received responses from many people telling us how to make the slides ourselves but only two people responded with actual suppliers of these slides. They were as follows;
1. "Molecular Probes, Inc. sells their "MultiSpeck Kit" Catalog # M-7900, consisting of three slides, each wiht two zones. List price is US$125." (thanks to Micheal Nesson). 2. "Polysciences make fluorescent beads already on slides Cat. # 24016 Confocal Microscopy-Multifluorescent Adjustment and Calibration Kit." (thanks to Jane Woodruff).
I hope this is of use to other people out there,
Sarah Ellis
Trescowthick Research Centre Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne 8006 Victoria Australia
At 16:37 31/08/98 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
width max picture width code
16 mm 13mm 110 35mm 24mm 135 35mm 28mm 126 single perforation 46mm 40mm 127 62mm 60mm 120 OR 620 OR 220 (double length)
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Has anyone out there any ideas on how best to visualise the protofilaments that make up the microtubule subfibres of respiratory cilia? We have tried tannic acid at 1% in the primary fix and at the final dehydration stage.
I would also be interested to hear from anyone with experience of immunogold labelling of the dynein arms.
I apologize for not incuding my address in my last posting. Here is th= e complete e-mail message.
Dear ICEM attendees:
We are looking for sands from around the world to support the GEMS/Proj= ect Micro elementary science program, MICROSCOPIC EXPLORATIONS. If anyone = would like to collect a handful of sand from Cancun for this project it would= be greatly appreciated. If you can send some sand please contact me first= so I can avoid receiving to many samples. The samples can be sent to:
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-3537 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com =
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I am trying to prepare virus crystals for HR-SEM. The crystals are 0.1mm or less in size and very delicate. Fixation and dehydration are not a problem. The problems come in the final drying and mounting for viewing.
I have tried drying by gradually replacing the 100% ETOH with Freon 113. However, the crystals float making them very hard to keep track of....also they tend to attach to the walls of the small tube and dry there. I need to get them onto something that can then be put into the SEM.
I normally will have only 2-3 of these crystals so cannot afford to have any lost in the process. They are very hard to see so I hesitate trying to put them onto filter paper...I am afraid they will get lost in the fibers.
I would appreciate any suggestions for ways to handle these guys.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
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I have a need to learn how to operate a Biorad confocal microscope -- the scope most available to me is model MRC 1000. I am based in St. Paul, MN, but will travel anywhere for a formal training session. Are such sessions available and, if so, where and when are they? Who do I contact?
Would prefer that you email me directly with your information. Thanks.
We use Epon (EMbed, Polybed, etc) for in situ embedding. Use only ethanol (not propylene oxide) for dehydration. Make sure absolute alcohol is dry by either using fresh bottle or storing it with molecular sieves. (Bake sieves every time bottle is emptied to dry. Let settle after adding ethanol for any sediment to settle out).
Not sure whether Maraglas can be processed this way. Suggestion for determining this to follow.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} Hi Folks, } I have been trying to use EMbed-812 for TEM on C.elegans. I am } using an ultracut } E, with a glass knife and I just can not seem to get any thin sections using } this resin.Poly/Bed 812 has worked fine, but is a little too viscous. I have } tried both medium and hard mixtures to no avail. Does anyone have any ideas, } I don't have access to a diamond knife. } } Thank You } } Patrick
Please be more specific about your problem. Are you getting tissue tearout, compression, poor infiltration, or what? Must you use EMbed-812 for some reason?
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
} Hi Folks, } I have been trying to use EMbed-812 for TEM on C.elegans. I am } using an ultracut } E, with a glass knife and I just can not seem to get any thin sections using } this resin.Poly/Bed 812 has worked fine, but is a little too viscous. I have } tried both medium and hard mixtures to no avail. Does anyone have any ideas, } I don't have access to a diamond knife. } } Thank You } } Patrick
Please be more specific about your problem. Are you getting tissue tearout, compression, poor infiltration, or what? Must you use EMbed-812 for some reason?
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
The New York Society of Experimental Microscopists 1998 Presidential Symposium and The Analytical Imaging Facility of the Albert Einstein College of Medicine
Present
"THE CELLULAR CELL: CREATION AND MAINTENANCE OF AN ORGANIZED CYTOPLASM"
Third Floor Lecture Hall Forchheimer Building, Albert Einstein College of Medicine
September 10, 1998 8:45 AM to 5:30 PM
8:45 AM Registration -- Coffee, 3rd Floor Conference Room
9:15 AM Welcome and Introduction Dr. John Condeelis, President, NYSEM
9:30 AM Dr. Tulle Hazelrigg, Columbia University "Getting the Message to its Destination: Localization of Bicoid mRNA in the Drosophila Oocyte"
10:15 AM Dr. John Condeelis, Albert Einstein College of Medicine "Reciprocal Regulation of mRNA Targeting and Actin Filament Dynamics"
11:00 AM Coffee Break with the Vendors, 3rd Floor Conference Room
11:30 AM Dr. Donald Ingber, Harvard Medical School "Tensegrity: The Mechanical Basis of Cellular Organization"
12:15 PM Dr. Mark Mooseker, Yale University "Myosin Superfamily of Actin Based Motors: Tails of Deafness, Blindness and Seizures"
1:00 PM Lunch at The Analytical Imaging Facility and Vendor Demonstrations, Room F641
3:00 PM Dr. Bruce Schnapp, Harvard Medical School "Biochemical Studies of the RNA Localization Machinery in Xenopus Oocytes"
3:45 PM Dr. Robert Singer, Albert Einstein College of Medicine "Molecular Biology through the Microscope: Intracellular Travels of an RNA"
4:30 PM Conclusion and Perspective Dr. Robert Singer
4:45 PM Open House wine and cheese reception at the Analytical Imaging Facility, Room F641
For additional information see: http://www.ca.aecom.yu.edu/aif/directions.htm
**************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue Bronx, NY 10461 ****************************************************************************
MMMS will host a meeting on Friday, October 9, 1998 at Purdue Universit= y in Lafayette, Indiana. The focus of the meeting will be Materials Scienc= e. Details will follow, but mark your calendars now!
Jane A. Fagerland, Ph.D. Dept. of Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064 (847) 935-0104 =
I am double labelling plant ovules with a view to observing nuclei and cytoskeleton within the cells of the embryo sac. I am using fixed tissue embedded in a low melting point wax (Steedmans wax) which is removed with alcohol prior to double labelling for tubulin (antibodies - FITC tag) and DNA (Hoechst).
Anti-fade agents (glycerol- PDA) and Citifluor work quite well for FITC but not at all for Hoechst, which fades and/or becomes non-specific throughout the tissue within 24 hours. A 'no-antifade' solution which is made up in glycerol (a mixture of biocarb buffers) and adjusted to pH 8.6 seems the best so far, but this is still not adequate to collect images with uv after 3-4 days.
I'd be grateful for any help
Meredith Wallwork (Dr) Department of Horticulture, Viticulture and Oenology Waite Campus University of Adelaide Sth Aust
Delicate specimen that defy handling during CPD and similar methods often are well preserved by a solvent drying method:
Line a glass Petrie dish with a double-layer of filter paper. Saturate the paper with chloroform (somebody ought to try how other solvents perform). Place a microscope slide onto the filter paper to keep the specimen off the paper. Sit the dehydrated, wet (absolute ethanol) specimen, which may be on a coverslip, a piece of mica etc onto the slide. Cover the Petrie dish and refrigerate for two days. Don't open Petrie dish until it has warmed to at least room temperature (incubator?) Proceed with sputter coating . . .
The method relies on very slow removal of the solvent saturated atmosphere within the dish and this lengthy time allows most of the remaining, trapped water molecules to leave the specimen without high pressures that distorts and shrinks specimens. I have used this method with microscopic nematodes; its not always perfect but for these critters its about the best means available. Certainly, its easy and avoids losing specimens. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Tuesday, 1 September 1998 23:52, Debby Sherman [SMTP:sherman-at-btny.purdue.edu] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } I am trying to prepare virus crystals for HR-SEM. The } crystals are } 0.1mm or less in size and very delicate. Fixation and } dehydration are not } a problem. The problems come in the final drying and } mounting for } viewing. } } I have tried drying by gradually replacing the 100% } ETOH with Freon } 113. However, the crystals float making them very hard to } keep track } of....also they tend to attach to the walls of the small } tube and dry there. I } need to get them onto something that can then be put into } the SEM. } } I normally will have only 2-3 of these crystals so } cannot afford to } have any lost in the process. They are very hard to see } so I hesitate } trying to put them onto filter paper...I am afraid they } will get lost in the } fibers. } } I would appreciate any suggestions for ways to handle } these guys. } } Debby Sherman, Manager Phone: 765-494-6666 } Microscopy Center in Agriculture FAX: 765-494-5896 } Dept. of Botany & Plant Pathology E-mail: } sherman-at-btny.purdue.edu } Purdue University } 1057 Whistler Building } West Lafayette, IN 47907-1057 }
Serge Oktyabrsky raised an interesting question about tapes suitable for cleaving HOPG (highly ordered pyrolytic graphite) to provide single crystal graphite support films for TEM. So far as I'm aware, there isn't one. Certainly the published references for this technique which I've seen use solvents which pose a considerable risk to laboratory workers and their neighbors.
If somebody comes up with an adhesive which adequately sticks to HOPG to cleave it well and then leaves no residue, I'd appreciate knowing about it.
Disclaimer: SPI Supplies sells HOPG, and we have an obvious interest in promoting its use. Furthermore, if we had a technique for reliable production of single crystal carbon support films from cleaved HOPG, we'd add them to our list of products.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
Have some sponge in here to thin section and eventually IEM. Any suggestions on getting rid of the spicules. They are of the silica variety and HF is not my first choice. Thanks
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Regarding a recent discussion about physical characteristics of hair: I just recieved a notice of a CRC book 'Atlas of Human Hair. Microscopic Characteristics', by Robert R. Ogle, Jr., and Michelle J. Fox. The book is targeted to forensic researchers and practitioners.
The book is due out Feb. 1999, and is priced at $99.95. ISBN: 0-8493-8134-7
I have deleted the original message, so cannot send this to the young student's contact person. If you would like the complete description of the book, contact me and I can fax (or mail) the ad to you.
Maureen Petersen
************************************************************************ Maureen Petersen Department of Plant Pathology 1453 Fifield Hall University of Florida
If you have access only to glass knives, you should immediately consider a mixed resin embedding. The "812s" used with DDSA and NMA are too hard on glass edges. Even with the best glass knife, relatively few thin sections can be cut. Not so with the mixed resin embedments. Once I spent 18 months sectioning tough muscle tissue with glass knives, and was able to get multitudinous sections before the glass edge wore out. It has been suggested to me (but I have not proof of this) that the molecular structure of NMA wears the glass edge too fast. Below is the formulation for this mixed resin which we use today frequently.
Polymerize 48 hours at 60C. Test your block. For a harder block, heat it at 95C for an hour, or put it back at 60C for another 24 hours. You may vary the cutting consistency of your block by adjusting (slightly) the dibutyl use 1/2% or 1%. I have used 2%, but found it too soft. It is a matter of preference and also your final decisions on what sort of sections you need - Silver? Gold? If you go to mixed resin embedding, be sure to mix the resin monomers very well. Keep them mixed. Do not let them just "sit". Keep your tissues on a rotator. Lenghten your embedding times. You are now dealing with the viscous Araldite. Use acteone as an intermediary and do several changes, making sure that all the acetone is out of the tissue. My phone # is 303-871-3026. Bye, Hildy
P.S. DMP-30 has gotten a lot of bad press lately. We have not run tests as to the substitution ratios for the better BDMA. MSA members cannot agree on the quantities either. quantities.
How about trying Nucleopore filters to trap your paticles. These filters are very smooth and I have used them successfully to entrap samples for SEM. There are are a variety of pore sizes available. I know they are available from SPI Supplies--probably other suppliers carry them also.
} Date: 01 Sep 98 08:51:42 -0500 } From: Debby Sherman {sherman-at-btny.purdue.edu} } Subject: virus crystals for SEM } To: "message to: MSA list" {microscopy-at-sparc5.microscopy.com} } X-Mailer: QuickMail Pro 1.5.3 (Mac) } Reply-To: Debby Sherman {sherman-at-btny.purdue.edu} } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have found that the SlowFade Light Antifade Kit from Molecular Probes does not quench shorter wavelength fluorophores such as DAPI and Hoechst. Their Prolong Kit has also produced superior results in some applications such as unfixed GFP expressing tissue. See their helpful handbook on line at: http://www.probes.com/handbook/toc.html go to Chapter 26.1 http://www.probes.com/handbook/ch26-1.html#ProLong
--just a happy customer
} Date: Wed, 2 Sep 1998 16:49:21 +0930 (CST) } From: Meredith Wallwork {mwallwor-at-waite.adelaide.edu.au} } Sender: Meredith Wallwork {mwallwor-at-waite.adelaide.edu.au} } Reply-To: Meredith Wallwork {mwallwor-at-waite.adelaide.edu.au} } Subject: re: antifade agents } To: Microscopy-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Perhaps the following correlation would be useful to the student: hair straightness and cross section are related. Straight hair is round in cross section, curly hair is more elliptical, and tightly curled hair is flatter still.
Leonard Corwin Fort Dodge Animal Health Princeton NJ
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Regarding a recent discussion about physical characteristics of hair: I just recieved a notice of a CRC book 'Atlas of Human Hair. Microscopic Characteristics', by Robert R. Ogle, Jr., and Michelle J. Fox. The book is targeted to forensic researchers and practitioners.
The book is due out Feb. 1999, and is priced at $99.95. ISBN: 0-8493-8134-7
I have deleted the original message, so cannot send this to the young student's contact person. If you would like the complete description of the book, contact me and I can fax (or mail) the ad to you.
Maureen Petersen
************************************************************************ Maureen Petersen Department of Plant Pathology 1453 Fifield Hall University of Florida
by heinlein.acpub.duke.edu (8.8.5/Duke-4.6.0) with ESMTP id PAA05118; Wed, 2 Sep 1998 15:50:52 -0400 (EDT) Received: (from saram-at-localhost) by soc11.acpub.duke.edu (8.8.5/Duke-4.4) id PAA05896; Wed, 2 Sep 1998 15:50:51 -0400 (EDT)
On Wed, 10 Jun 1998, Ellis, Sarah wrote:
} Date: Wed, 10 Jun 1998 13:45:39 +1000 } From: Ellis, Sarah {s.ellis-at-pmci.unimelb.edu.au} } To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} , } Microscopy-at-sparc5.microscopy.com } Subject: Fixing and processing cell colonies grown between agar sheets. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi! } } Can you help us? A student is growing very small colonies of cells and } she wishes to view their ultrastructure. There are about 50 cells per } colony and about 4 colonies per 3cm polystyrene petri dish. Our problem } is that the colonies are growing between two layers of agar. The bottom } layer is 0.5% agar in PBS and the top layer is 0.33% agar in PBS. The } colonies break up and float away during processing and the agar just } moves around. The colonies are not attached/embedded in the agar. } We have tried (1)cutting around the colonies and sucking the whole lot } up (agar + colony) and treating it as a pellet but the cells are } impossible to find in the resin, and (2)we have tried to just gently } fix the mass and process it as a whole but we end up with no cells as } the colony breaks up and the cells disperse. } Is there anyone out there who could offer a suggestion. } Thanks } } Sarah Ellis } } } Research Division } Peter MacCallum Cancer Institute } Locked Bag #1 } A'Beckett Street } Melbourne, Victoria 3000 } Australia } } Phone 61-3-9656 1244 } Fax 61-3-96561411 } Email s.ellis-at-pmci.unimelb.edu.au {mailto:s.ellis-at-pmci.unimelb.edu.au} } I suggest:
Keep agar layers as thin as possible (barely covering the plate and then the inoculated cells. It will be only a couple mm thick.
Infiltrate/embed the whole agar layer in situ as you would for an adherrent culture.
Use a microscope to locate the cells, circling the colony with a thin magic marker on the bottom of the plate.
After baking and before you peel up the agar/resin layer, transfer the circle to the upper surface of the layer. You can even put this circle of resin back under a microscope to see the cells which will be slightly brown from the osmium.
THEN cut out the colony and glue onto a blank stub.
We have done this successfully with very small colonies.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We trialled a number of antifade solotions, both commercial and DIY last year . Far and away the best for our purposes was Molecular Probes Prolong, Also the most expensive, but you get what you pay for, I suggest you give it a go. Pete Smith AgResearch Wallaceville Upper Hutt New Zealand
I read some of the other responses to your problem and I have used the Prolong Anti-Fade from Molecular Probes and love it. However, we did come up with a very cheap media that has worked very well for double and triple labels in cojuction with a DAPI stain.
70% glycerol 25% .5M tris pH 9.0 5% N-propyl gallate
Heat in a boiling water bath to dissolve the n-propyl gallate. Cool and pH to 7.4 Keep light tight at 4 degrees C. Use as little as neccessary to cover the coverslip.
Bob Derm Imaging Center U of W Seattle
On Wed, 2 Sep 1998, Meredith Wallwork wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } I am double labelling plant ovules with a view to observing nuclei and } cytoskeleton within the cells of the embryo sac. I am using fixed tissue } embedded in a low melting point wax (Steedmans wax) which is removed with } alcohol prior to double labelling for tubulin (antibodies - FITC tag) and } DNA (Hoechst). } } Anti-fade agents (glycerol- PDA) and Citifluor work quite well for FITC } but not at all for Hoechst, which fades and/or becomes non-specific } throughout the tissue within 24 hours. A 'no-antifade' solution which } is made up in glycerol (a mixture of biocarb buffers) and adjusted to pH } 8.6 seems the best so far, but this is still not adequate to collect } images with uv after 3-4 days. } } I'd be grateful for any help } } Meredith Wallwork (Dr) } Department of Horticulture, Viticulture and Oenology } Waite Campus } University of Adelaide } Sth Aust } } }
When I was in second year, a long time ago I did a Scanning Tunneling Microscopy (STM) exoeriment as part of my materials lab techniques. One of the samples that we studied was HOPG, and we used normal sticky tape to cleave a new surface. The technique was to press a piece of tape onto the HOPG and then just rip it off.
Perhaps you could remove the film on the tape by dissolving the tape adhesive in ethanol, float off the graphite film and then dry it by baking gently. If the gods are smiling, hopefully it will be a single crystal graphite support film.
Don't know if it helps but thats my 2 cents worth!
I have a client that I am assisting in finding a used SEM for a new metallurgical lab. We have been looking at a couple, but I thought there might be some citizens of this list that may be looking to dispose of some older equipment for a good price. LaB6 would be nice but not necessary. EDS also desired, preferably thin window.
Please respond by email if you have an instrument you'd like to sell. Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Stowe and Robinson report about reducing beam scattering in conventional Low Vacuum SEM's (Scanning, Vol. 20, 57-60). Are there any experiences using Helium in an ESEM from Electroscan or Philips with a special ESEM-detector? Is the ionization efficiency high enough to get a good performance for amplifying the electrons coming from the sample? How is the image quality compared to e.g. water vapor?
Kind regards
Rainer Ziel ------------------------------------------------------------- Dipl.-Phys. Rainer Ziel Akzo Nobel Central Research ACR-O/RMG-EM D-63784 Obernburg Germany
As I remember from about five years ago, a student with whom I shared a lab was cleaving HOPG for a STM resolution test specimen by using some form of sticky tape. Dr Mark Aindow was his (and my) PhD supervisor and he can be reached on m.aindow-at-bham.ac.uk, perhaps he would remember the details as he oversees the STM/AFM facility. I was under the impression, however, that this was not some new technique invented by them but something that had been done previously by other STM researchers.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
I work with Atriplex nummularia and I want to examine epidermal cell patterns. This specie has a proffusion of glandular hairs in both surface= s. I read in a paper something about use cellulose acetate film with acetone to make impressions of the leaf.=20
Someone know something different and simplest about this subject? I also need to know what to do to take off these trichomes. Thank in advance.
Rejane
Rejane Magalh=E3es Pimentel Galindo =20 ggalindo-at-elogica.com.br Universidade Federal Rural de Pernambuco Av. Boa Viagem, 6592/602 FAX: 55 (081) 4416177 51130-000, Recife, Pernambuco, Brasil
Sory, I forgot the final of this message. Here it is complete.
I work with Atriplex nummularia and I want to examine epidermal cell } patterns. This specie has a proffusion of glandular hairs in both surfaces. } I read in a paper something about use cellulose acetate film with aceto= ne } to make impressions of the leaf.=20 } =20 } Someone know something different and simplest about this subject? } I also need to know what to do to take off these trichomes.
Aside this, I want to obtain the vein impressions; I need a simple method to highlight the leaf venation. } Thank in advance. } =20 } Rejane
Rejane Magalh=E3es Pimentel Galindo =20 ggalindo-at-elogica.com.br Universidade Federal Rural de Pernambuco Av. Boa Viagem, 6592/602 FAX: 55 (081) 4416177 51130-000, Recife, Pernambuco, Brasil
Dear listers, we need to do service on JEM 2010 that I suppose is normaly done by = service people as there is nothing about it in the manual. Namely that = is dismounting and clearning of sample lock. Due to some reasons we are = not able to call for JEOL ingineer and have to do all by ourselfs. Does = anyone know of service (persons) whom we could ask for advice? Schemes = and drawings would help greatly.
Before we begin a thread on how to cleave HOPG, may we all please recall that the question was how to cleave it without leaving any tape residue. To cleave using almost any tape is a matter of technique. To cleave without leaving any tape residue is the problem for which the original writer (Serge Oktyabrsky) is seeking a solution.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
Here is how we were able to get good 'screen shoot' color slides:
Film: Kodak Ektachrome color slide film; ASA 100
Settings: Camera -- Nikon with tripod and cable shutter release; f/2.8 (wide open); 1/2 and 1 sec. No flters.
Computer screen settings: At the 1 sec. exposure, dimming the brightness/contrast on the screen slightly, gave good results. Recommend shooting at 1/2 sec. with a screen appearance good 'to the eye' and 1 sec. with slight screen dimming. Take these two shots on each subject for a choice (often both slides are usable).
Room conditions: Total darkness -- no room lights; close door. This eliminates any glare on computer screen by room light.
Hope this is helpful. Please ask about anything I may have overlooked. Gerald Harrison ======================================================================== At 04:29 AM 9/3/98 -0700, you wrote:
} } I would like to photograph my multi-sync computer monitor screen using } 35mm color slide film. } } Assuming it is worth a try... does anyone have suggestions regarding: } 1)film type 2)shutter speed 3)filters? } } Thank you. } } Bart Cannon } Cannon Microprobe } }
Greetings. Does anyone know how long you can keep Formvar in solution? I need to make perfect films. I have a lot of Formvar in Ethylene Dichloride, but it has been on the shelf unopened since 1990. Can I use it?
} } } I would like to photograph my multi-sync computer monitor screen using } 35mm color slide film. } } Assuming it is worth a try... does anyone have suggestions regarding: } 1)film type 2)shutter speed 3)filters? } } ...
The hardest part may be capturing the dynamic range of the monitor ... you may want to reduce its contrast a bit ... but its difficult to know how much.
Depending on your subject matter ... you may also want to create a standard gray image to meter on ... gray level (R,G,B) = 70 ought to be pretty close. An equivelent and/or comparison can be made by metering "white" with an ASA setting = 1/5 of the film speed (... a copy stand trick ...)
The lens should be a portrait type ... e.g., 100-135mm
The shutter speed should capture many screen refreshes ... e.g., 1/8 sec ... and turn the room lights off.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
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Debby,
How about cryo-SEM technique? The virus crystals in suspension are absorbed on substrata, fast frozen, partial freeze-dried, cryo-coated with a thin layer of metal, then viewed in a cryo-SEM. I have used this protocol for many years to look at individual viral particle by our high-resolution cryo-SEM.
Ya Chen
Ya Chen
======================================================================== \ / Integrated Microscopy Resource (IMR)-- \ / __ a NIH Biomedical Research Resource TEL: 608-263-8481 \/ / / University of Wisconsin-Madison FAX: 608-265-4076 / / / 1675 Observatory Drive #159 / /__/_ Madison, WI 53706 Email: ychen14-at-facstaff.wisc.edu ======================================================================== IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr/home.htm
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RE} Anybody knows about the_ 9/3/98 10:18 AM Dear Gehoon-at-plaza1....,
I am not sure what your real question is. However, you may want to be awa= re that phase contrast is major contrast technique used in light microsco= py. The technique was developed for telescopes by Frits Zernike (1934) an= d later applied to the microscope by Kohler and Loos at Zeiss.
Phase contrast provides an efficient method for enhancing the contrast of= specimens whose refractive indices are very similar and/ or transparent = (or nearly so). The technique does not apprciable lessen the resolution o= f the optics. The technique is especially used on living cells.
I hope this abbreviated answer will serve you. You may want to visit the = library and pick up any modern light microscopy textbook in the later hal= f of the 20th Century for a more detailed explaination.
john shane
-------------------------------------- the internet for 5 hours, and still don't have the answer. If can anybo= dy help me, please e-mail me at gehoon-at-plaza1.snu.ac.kr =0A {!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} =0A {HTML} =0A {HEAD} =0A =0A {META content=3Dtext/html;charset=3Dks_c_5601-1987 http-equiv=3DConten= t-Type} =0A {META content=3D'"MSHTML 5.00.0518.7"' name=3DGENERATOR} =0A {/HEAD} =0A {BODY bgColor=3D#ffffff} =0A {DIV} {FONT color=3D#000000 face=3D size=3D2} I am a freshman studying d= entistry in=20 =0ASeoul, Korea. Yesterday my professor gave us a question which we= were=20 =0Asupposed to answer via e-mail as quick as we could. The problem = was=20 =0A"Why is there the 'phase contrast' in PCM?" I looked u= p all the=20 =0Areferences available to me, and searched all over the internet for 5 h= ours, and=20 =0Astill don't have the answer. If can anybody help me, please e-ma= il me at=20 =0A {A=20 =0Ahref=3D"mailto:gehoon-at-plaza1.snu.ac.kr"} gehoon-at-plaza1.snu.ac.kr {/A} {/F= ONT} {/DIV} =0A {/BODY} {/HTML} =0A
by bubba.NMSU.Edu (8.9.0/8.9.0) with ESMTP id JAA17863; Thu, 3 Sep 1998 09:28:10 -0600 (MDT) Received: from nestor.NMSU.Edu (nestor.NMSU.Edu [128.123.34.146]) by dns1.NMSU.Edu (8.9.0/8.9.0) with ESMTP id JAA11839; Thu, 3 Sep 1998 09:27:04 -0600 (MDT) Received: from microscope.nmsu.edu (microscope.NMSU.Edu [128.123.5.85]) by nestor.NMSU.Edu (8.8.6/8.7) with SMTP id JAA139504; Thu, 3 Sep 1998 09:27:05 -0600 Message-Id: {3.0.6.32.19980903092600.007aa710-at-cnmailsvr.nmsu.edu} X-Sender: rtindell-at-cnmailsvr.nmsu.edu X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32)
At 04:29 AM 9/3/98 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Bart,
Although it's bean a while since I've done this, I believe that most computer screens will photograph fairly well on regular daylight slide film (somebody please correct me if I'm wrong), which gives you a pretty wide choice. The slower speed films give you finer grain. A good bet might be one of the 100 speed Fujichromes or Ektachromes.
As to shutter speed, you will want to use a shutter speed slower than the screen refresh rate to avoid bands. You'd be safe with 1/30 of a second or slower for almost any screen, including your home tv.
Good luck.
Randy
} Randy Tindall Electron Microscope Laboratory Box 3EML--Biology New Mexico State University Las Cruces, NM 88003
Transmission microscopy in some contexts might be thought of as the taking of transmitted wave pictures of foils at high magnification -- a kind of seriously enlarging xerox machine that only works for "transparencies". Some microscopes in the past, I believe, may have been designed with this perspective in mind.
The kinds of electron scattering visitation, specimen modification, and analysis, done especially with crystalline specimens, is of a different sort. It rather seems that the microscope is simply a kind of "suit" that you can put on, and that allows one to visit places as a nano-human, and to perform various kinds of electron scattering and data recording activities. Different people, when they visit the tiny places accessible with such "suits", do very different things when they get there, and bring back different stories, much as visitors to the same south sea island might go to the same place but have different experiences as well. Moreover, unlike xerox machines, a suit is merely an extension of a human, and although it may have some personality, it does little by itself.
In this latter spirit, I offer the alternative meaning for the acronym "TEM" given in the subject to this message. For a bit further development of this metaphor, you might enjoy the announcement by this same name linked to our TEM course page* this semester.
As long as you keep it in the dark, it can last a long time. When exposed to light, you tend to form HCl molecules in the dichlorethane weakening the formvar.
Henk
At 10:46 AM 9/3/98 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I can't give you the exact answers you are looking for, but hopefully the following can point you in the right direction. At the '99 MSA meeting in Atlanta, there was a session on ESEM. Several papers were given by the group from the Cavendish Laboratory, especially one on SEM at freezer temperatures by A.L. Fletcher, ... & A.M. McDonald. In an attempt to maintain the proper humidity without icing or thawing, this group used other gases, and determined that N2 was minimally acceptable in terms of obtaining desired imaging SEs. Discussion following the paper included comments on the scattering and SE generation that could be expected from lower atomic number gases, including He. I would suggest your contacting this group. Additionally, I would recommend that you do a literature search for the work of K. Rudiger-Peters and the various publications he has on the physical characteristics and SE generation in the ESEM. I think most of those papers were in the journal Scanning. The principal author on the ESEM is of course Danilatos, and his publications span from 1982 to 1990. Since I do not have ready access to those publications (everything is in boxes, no longer arranged as per my reference manager database) I can't give you figures or exact references even, but I believe that he did publish on different gases. Hope this helps.
Roger Moretz Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
} -----Original Message----- } From: Ziel, R. (Rainer) [SMTP:Rainer.Ziel-at-akzonobel.com] } Sent: Thursday, September 03, 1998 2:43 AM } To: 'MSA' } Subject: ESEM: Use of Helium Gas } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Stowe and Robinson report about reducing beam scattering in } conventional } Low Vacuum SEM's (Scanning, Vol. 20, 57-60). Are there any experiences } using Helium in an ESEM from Electroscan or Philips with a special } ESEM-detector? Is the ionization efficiency high enough to get a good } performance for amplifying the electrons coming from the sample? How } is } the image quality compared to e.g. water vapor? } } Kind regards } } Rainer Ziel } ------------------------------------------------------------- } Dipl.-Phys. Rainer Ziel } Akzo Nobel Central Research } ACR-O/RMG-EM } D-63784 Obernburg } Germany } } Tel: (06022) 81-2645 } Fax: (06022) 81-2896 } E-mail: Rainer.Ziel-at-AkzoNobel.com }
SE, You can also make bead slides, bought from molecular probes in many sizes and emissions. A little poly-l-lysine to make them stick to the coverslip and you are good to go.
At 09:26 AM 9/3/98 -0600, Tindall wrote: } Although it's bean a while since I've done this, I believe that most } computer screens will photograph fairly well on regular daylight slide film } (somebody please correct me if I'm wrong), which gives you a pretty wide } choice. The slower speed films give you finer grain. A good bet might be } one of the 100 speed Fujichromes or Ektachromes. } } As to shutter speed, you will want to use a shutter speed slower than the } screen refresh rate to avoid bands. You'd be safe with 1/30 of a second or } slower for almost any screen, including your home tv.
I don't think 1/30th will be long enough, especially for TV's. They scan at 60 frames per second but only refresh half the lines per pass, so it takes 1/30th of a second to get one whole image. Now if the camera shutter is exactly 1/30th second, then you would get one and only one complete pass.
We tried a similar exercise on one of our EDS monitors years ago and used too fast a speed. I think I figured we captured 4 and some scans. We could see a brighter band on the slide where 5 frames had been scanned compared to where only 4 frames had been scanned.
So longer is better, and you will probably want to use a tripod for making sure you have the same setup between shots anyway.
I used my 60mm macro (Nikon calls macro lenses micros), and just trusted my light meter in the camera, and used it on program mode, for perfect results with the Nikon F4 -35mm camera. With the macro lens, you can get close enough to fill the frame with the image of the computer screen. You just have to be careful to turn off the room lights to avoid glare, and have the camera parallel with the screen. No filters are necessary, and you can use ordinary daylight slide film.
} ---------- } From: shAf[SMTP:mshaf-at-darkwing.uoregon.edu] } Sent: 3 September, 1998 10:08 } To: cannonmp-at-accessone.com; MSA Listserver } Subject: RE: Photos of Comptr Scrn } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We wish to buy used AFM systems and components made by Digital Instruments:
NanoScope II NanoScope III Contact mode or Multimode AFM or Dimension AFM AFM base AFM scanners AFM optical head Accessories and tools used with NanoScope equipment, including fiberoptic illuminator, stand for monocular microscope, vibration isolation pad, acoustic shroud.
Example: If you originally had a NanoScope II AFM and upgraded to a NanoScope III AFM to do TappingMode, you may still have a contact mode AFM base and optical head that you are not using.
If you have equipment to sell, or if you know someone who may have such equipment, please contact me offline (directly) at the address shown below. Do not respond to this discussion group.
Thank you.
Don Chernoff
Advanced Surface Microscopy, Inc. E-Mail: asm-at-indy.net 6009 KNYGHTON RD. Voice: 317-251-1364 INDIANAPOLIS IN 46220 Toll free: 800-374-8557 (in USA) web: http://www.a1.com/asm Fax: 317-254-8690 (note: "a1"= letter "a", numeral "1")
Dear Bart, } } I would like to photograph my multi-sync computer monitor screen using } 35mm color slide film. } } Assuming it is worth a try... does anyone have suggestions regarding: } 1)film type 2)shutter speed 3)filters? } I have been successful at photographing monitors; however, there are distortions due to the difference between the camera-screen distances to the center or edge of the screen. Bearing this in mind, one would like to use a high f-number for a large depth-of-field. I used (1) ektachrome 100 professional film, (2) the auto-exposure setting (an exposure much longer than the refresh time is good so that many frames contribute to the image), and (3) no filter. If distortions are not acceptable, try a tele- photo lens, but be prepared for a very long exposure. Good luck. Yours, Bill Tivol
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings. } Does anyone know how long you can keep Formvar in solution? I need to make } perfect films. I have a lot of Formvar in Ethylene Dichloride, but it has } been on the shelf unopened since 1990. Can I use it? } } Sally Shrom } } } Hi! Formvar solutions deterioate due to atmospheric moisture and age. This causes holes. If not holes, then "measles" (light areas in the film). It is not worth ruining sections because one has used old Formvar solution. Buy or make new from powder. Make a few films and check in the TEM for stability, thickness, etc., before making a thousand. Bye, Hildy
Sally Shrom wrote: } } } Greetings. } Does anyone know how long you can keep Formvar in solution? I need to make } perfect films. I have a lot of Formvar in Ethylene Dichloride, but it has } been on the shelf unopened since 1990. Can I use it? } } Sally Shrom Hi Sally,
I use Formvar in Cloroform solution in a well stopped glass ( 100 ml, 0.25%). I am using the solution at 1 year and obtained excellent films. With a pen, mark the level of the solution and if it evapore, refill with Cloroform. The Butvar give excellent films too and it easy to make very fine films. Rinaldo Pires dos Santos UFRGS - Dept. of Botany - Lab. of Plant Anatomy Porto Alegre - RS - Brazil e-mail: rinaldop-at-botanica.ufrgs.br
You might consider the advantages of a digital screen capture program. I use one called Capture for the Macintosh. It saves at the screen image resolution and allows a variety of formats (PICT, TIFF, etc.). Open it in Photoshop, clip, adjust colors, add annotation, and print on you dye-sub or ink-jet printer. No film to mess with, no chemicals, no fiddling with exposures, no wait. What could be sweeter?
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Bart, I have often photographed images on the computer screen with good results. I have found that Kodak film tends to give slides that are too blue. However Polaroid Presentation Chrome really gives you what you see. You get very good greyscale and colors are realistic as well. We use a 60mm lens and photograph from a few feet from the monitor in order to get the entire monitor screen to fill the field. However, the further away you can get from the monitor, the less your image will be affected by screen curviture. Do try to use as flat a monitor as possible. I normally will shoot an exposure series by changing the f-stop using the camera automatic exposure meter as my guide. You must have a very high resolution monitor to really produce good slides and usually an electronic slide maker will do a bit better. However, this method works well and is quite fast to do once you get the hang of setting it up. Do take the time to level the monitor so the screen is parallel with the camera, use a sturdy tripod to hold the camera and use a cable release or automatic shutter delay (if your camera is so equipped). Good luck,
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------
I would like to photograph my multi-sync computer monitor screen using 35mm color slide film.
Assuming it is worth a try... does anyone have suggestions regarding: 1)film type 2)shutter speed 3)filters?
Thank you.
Bart Cannon Cannon Microprobe
RFC822 header -----------------------------------
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At 04:29 3/09/98 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
An ordinary 50 mm lens can focus close enough to use. At 1/4 second you need a tripod or some other way to steady the cameera
Its important to keep the room dark to minimise reflections from the front of the screen. Polaroid used to sell a handy plastic hood which pressed up against the screen and simultaneously eliminated reflections and steadied the camera.
We never used filters. Color is very subjective, especially when being shown as slides.
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
On Mon, 10 Aug 1998, Dr. Gary Faulkner, Electron Microscopy Unit wrote:
} Date: Mon, 10 Aug 1998 11:47:02 AST } From: Dr. Gary Faulkner, Electron Microscopy Unit {gfaulkner-at-tupdean1.med.dal.ca} } To: Microscopy-at-sparc5.microscopy.com } Subject: EM Decline } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } While my EM Unit caters strictly to research, I am often asked } whether I would agree that the usefulness of the EM in the } clinical setting is in a major decline. It is pointed out } that diagnostic EM, especially in the area of tumors, is rapidly } being replaced by immunocytochemistry at the LM level. In addition, } tumor cells do not as a class exhibit ultrastructural changes that } can tell us much in the way of significant information anyway. I was } wondering if you might have an opinion on this observation or a } reference(s) where I could check this out. } } Many thanks, } } Gary } } gary.faulkner-at-dal.ca } I can only answer for our situation:
My Surgical Pathology EM unit is holding steady at around 350-400 cases/year--muscle, nerve, heart, kidney, tumors--all thin sections.
My EM Diagnostic Virology unit does about 1000 samples per year, steady over the last couple of years, and July-August, 1998 same as last year. This case load is mostly (90%) negative stains of fluid samples processed by negative staining; 10% is thin sectioning of tissues.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} I don't think 1/30th will be long enough, especially for TV's. They scan at } 60 frames per second but only refresh half the lines per pass, so it takes } 1/30th of a second to get one whole image. Now if the camera shutter is } exactly 1/30th second, then you would get one and only one complete pass. } } We tried a similar exercise on one of our EDS monitors years ago and used } too fast a speed. I think I figured we captured 4 and some scans. We could } see a brighter band on the slide where 5 frames had been scanned compared to } where only 4 frames had been scanned. } } So longer is better, and you will probably want to use a tripod for making } sure you have the same setup between shots anyway. } } Warren } } Warren,
Thanks for the correction. I was unaware of the "every other line" refresh rate, and it's a valuable piece of information. I have never had any problems photographing screens at 1/30 sec., but I haven't done this recently. Next time I'll use 1/15 or slower.
Randy
Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
While on the topic of photographing monitor screens - If you focus the camera properly and focus the negative properly you can often see the scan lines quite strongly on the final print. Although it may go against the grain, slightly defocus the image on the negative and again on the print and the final image looks better for the blurring of the scan lines.
We take 0.5 and 1 sec exposures f 5.6 or 8 to get the best image from our B+W monitors.
Good luck, Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Rejane- Replicating with various plastic films is easy. Apply a drop or two of the recommended solvent (acetone or=20 methyl acetate) onto the surface. Cover with a small rectangle of replicating film, this=20 should have a notch near a corner to identify "specimen=20 side". Apply for a moment gentle pressure, perhaps using a=20 microscope slide. Leave to dry for five minutes. Pull the replica off. Angle coating (say 30 degrees) with metal helps to=20 visualise depths for light microscopy, whereas for SEM,=20 sputter coating is preferable. For TEM a secondary replica=20 of carbon and metal would be required, but that is another=20 story.
The replica would probably remove the plant hairs and a=20 subsequently applied replica may be preferred. Also, many=20 leaf surfaces are covered with wax. This may be removed by=20 flushing the leaf prior to replication with xylene. Rejane, if you require more details email me direct. Jim Darley ProSciTech Microscopy=20 PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au=20 *****
On Thursday, 3 September 1998 20:01, Rejane Magalh=E3es=20 Pimentel Galindo [SMTP:ggalindo-at-elogica.com.br] wrote: } } I work with Atriplex nummularia and I want to examine } epidermal cell } patterns. This specie has a proffusion of glandular } hairs in both surfaces. } } I read in a paper something about use cellulose acetate } } film with acetone } } to make impressions of the leaf. } } } } Someone know something different and simplest about=20 this } } subject? } } I also need to know what to do to take off these } } trichomes. } } Aside this, I want to obtain the vein impressions; I need } a simple method } to highlight the leaf venation. } } Thank in advance.
} Rejane Magalh=E3es Pimentel Galindo } ggalindo-at-elogica.com.br } Universidade Federal Rural de Pernambuco } Av. Boa Viagem, 6592/602 } FAX: 55 (081) 4416177 } 51130-000, Recife, Pernambuco, Brasil
Several months ago someone posted a message about difficulties they were having with quantifying synapses in LM preparations. I'd be grateful if this person would contact me offline. I've tried to find this message in the archives, without success, but that could be from my inexperience with searching there. Thanks,
I am in the market for a digital camera that can fulfill a wide range of objectives, including fluorescence imaging of fura. The experiments would probably be time-varying, which would limit exposures to ~1/2 second at best. Judging by the fact that the systems I have seen use intensified CCD (ICCD) cameras, my impression is that fura produces a very weak signal. Does anyone know if there are any non-intensified cameras (eg back-illuminated) capable of the same performance, or are ICCD cameras the only way to go?
Thanks
Eric
Eric Johnston Department of Bioengineering University of Pennsylvania 120 Hayden Hall 3320 Smith Walk Philadelphia, PA 19104-6392 215-898-1958 (F) 215-573-2071 ericdj-at-seas.upenn.edu
Prepares biological specimens for microscopic observation and analysis, using a variety of laboratory techniques and procedures. Assists in maintaining the laboratory and work environment at Bessey Microscopy Facility. Requirements: Bachelor's degree in a biological science discipline with knowledge in general chemistry, quantitative analysis, general physics, the use and general maintenance of light and electron microscopes, ancillary equipment, and all phases of handling and preparing biological specimens. Must be able to work as a part of a team, to communicate effectively with the clients, and to develop a work schedule that fits the needs of the BMF. Preferred: Master's degree in a biological science discipline; two years of laboratory experience, which may include academic courses; and practical experiences in the use of optical light microscopes, scanning and transmission electron microscopes, rotary and ultra-microtomes; darkroom experience; and knowledge of computers, networking and digital imaging; BEMT Certification (MSA). Send three (3) letters of reference and resume to Dr. Harry T. Horner, Bessey Microscopy Facility, Room 3A Bessey Hall, Iowa State University, Ames, IA 50011-1020. Fax 515.294.1337 or hth-at-iastate.edu ISU is an EO/AA employer
Tracey M. Pepper Supervisor Bessey Microscopy Facility Iowa State University Ames, IA 50011-1020 Phone: 515.294.3872 FAX: 515.294.1337 email: tpepper-at-iastate.edu
I am helping a local high school set up a scanning electron microscope for their advanced placement physics program.
Rather than draft a new course, I would like to know if anyone has a suitable teaching materials for an introduction to electron microscopy. This might include a course outline and names of textbooks.
Also, the school could use sputter coater.
Thanks in advance for help.
Alan Stone ASTON Metallurgical Services 4201 N Ravenswood Ave Chicago, IL 60613 Phone 773/528-9830
A uniform Polystyrene Latex Standard is available from SPI supplies USA, part # 02709-AB, having following specs:
Particle diameter: 0.105 um Uncertainty : 2 nm
This std. can be used to calibrate SEM. I would recommend to first calibrate the micron bar on the SEM using this std. (0.105 um particle/s). Size of the micron bar can be precisely adjusted by varying a potentiometer, provided at the display unit (with most SEM's). End result should be observed on the micrograph for comparison with actual sample. Because usually the SEM viewing screens display oversized image and have a multiplication factor to the actual magnification, while on the micrograph you get direct magnification.
Hope my 2 cents will help. Best of luck.
Zia ur Rahman E/M Engineer University of Central Florida
( Getting old is when a narrow waist and broad mind change places )
At 08:26 AM 9/4/1998 -0500, Martin Klein wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In a message dated 98-09-04 19:17:59 EDT, you write:
{ { Martin:
A uniform Polystyrene Latex Standard is available from SPI supplies USA, part # 02709-AB, having following specs:
Particle diameter: 0.105 um Uncertainty : 2 nm } } Although the uncertainty relating to particle diameter might be 2nm one must also consider possible additional error due to shrinkage of the spheres, both during sample preparation and when subjected to high vacuum and the SEM beam.
} I am helping a local high school set up a scanning electron microscope for
} their advanced placement physics program.
}
} Rather than draft a new course, I would like to know if anyone has a
} suitable teaching materials for an introduction to electron microscopy.
} This might include a course outline and names of textbooks.
}
} Also, the school could use sputter coater.
}
} Thanks in advance for help.
}
} Alan Stone
} ASTON Metallurgical Services
} 4201 N Ravenswood Ave
} Chicago, IL 60613
} Phone 773/528-9830
Alan,
One of the best places to start re:both textbook and course materials, is "Scanning Electron Microscopy: A Student's Handbook" (Postek, et. al), available through Ladd Research Assoc, Williston VT (802-878-6711). I believe they also have a website and are probably listed at the MicroWorld metasite at www.mrwn.com.
Rejane Magalhăes Pimentel Galindo wrote: =============================================== I work with Atriplex nummularia and I want to examine epidermal cell patterns. This specie has a proffusion of glandular hairs in both surfaces. I read in a paper something about use cellulose acetate film with acetone to make impressions of the leaf.
Someone know something different and simplest about this subject? I also need to know what to do to take off these trichomes.
Aside this, I want to obtain the vein impressions; I need a simple method to highlight the leaf venation. =================================================== The cellulose acetate material is generally provided by those offering EM consumables as "replicating tape" or "replicating sheets". Not all cellulose acetate is the same of course, but the grades offered from most sources tends to be acceptable for surface replication. You can find information about these materials and their use on our website.
However, the use of acetone might not be desirable since it is a solvent for the oils present on the leaf surface. Also, the dried cellulose acetate film tends to be stiff and brittle and tends to "pull off" some of the fine features from the sample making the "impression" difficult to interpret. It is also an "inverted" or "negative" image of the surface, something sometimes not so easy to interpret.
Another alternative would be our own SPI Wet Replica Kit. It was originally developed for replicating human skin in vivo and other non-dry surfaces. It too can be found on our website. It is a silicone resin based system and hence will not dissolve. It is cured with a very fast acting catalyst. The silicone really does not want to "wet" the surface and therefor the adhesion , and therefore also the tendency to pull off fine features is much less. But at this point, again you do end up with a "negative" replica.
The kit comes with another material (polyolefin) that is used to "replicate the replica" thereby generating a positive replica which should look just like the original. This now (vacuum) inert sample can be treated like any other SEM sample. This approach also makes possible the following of the same identical area, as a function of time, almost like a time-lapse- photography effect but at the SEM level.
However, like with all else in life, there are trade offs, the main one here being that above about 700X in an SEM you start to see structure from the replicating system itself. So if the features you are wanting to see can be seen below about 700X, this approach might work for you.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I have been interested in FTIR microscopic spectroscopy. I have worked in IR-transmission, reflectance and absoption(?-excuse me, this mode always confuses me). Most of my work was with metals and oxides,ocasionally organics. Microscopic spectroscopy,of course, has edge and aperature shift effects. But most microscopic systems I have tried to use have seemed to have have chronic incurable alignments and operation problems. I occasionally hear a success story and several papers published, but I've most frequently heard the same chronic failure stories. Can anyone recall similar situations and a cure? Also does anyone know a list server dedicated to FTIR microscopic spectroscopy? Please address any negative info to my Email or respond generically and technically on this server. The possibilities seem very exciting in FTIR microscopic spectroscopy, but I'm afraid there is a larger group of similar problems out there. If this is the case, I'd like to work toward a cure and notify all who need and the server members. Jeff Day Mesquite, Texas Email: WA5EKH-at-Juno.Com
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
I have been interested in FTIR microscopic spectroscopy. I have worked in IR-transmission, reflectance and absoption(?-excuse me, this mode always confuses me). Most of my work was with metals and oxides,occasionally organics. Microscopic spectroscopy,of course, has edge and aperture shift effects. But most microscopic systems I have tried to use have seemed to have had chronic incurable alignments and operation problems. I occasionally hear a success story and several papers published, but I've most frequently heard the same chronic failure stories. Can anyone recall similar situations and a cure? Also does anyone know a list server dedicated to FTIR microscopic spectroscopy? Please address any negative info to my Email or respond generically and technically on this server. The possibilities seem very exciting in FTIR microscopic spectroscopy, but I'm afraid there is a larger group of similar problems out there. If this is the case, I'd like to work toward a cure and notify all who need and the server members. Jeff Day Mesquite, Texas Email: WA5EKH-at-Juno.Com
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
In a message dated 9/5/98 6:21:31 PM EST, cgarber-at-2spi.com writes:
} =============================================== } I work with Atriplex nummularia and I want to examine epidermal cell } patterns. This specie has a proffusion of glandular hairs in both } surfaces. I read in a paper something about use cellulose acetate film } with acetone to make impressions of the leaf.
} } Greetings. } } Does anyone know how long you can keep Formvar in solution? I need to make } } perfect films. I have a lot of Formvar in Ethylene Dichloride, but it has } } been on the shelf unopened since 1990. Can I use it? } } } } Sally Shrom } } } } } } } Hi! } Formvar solutions deterioate due to atmospheric moisture and age. This } causes holes. If not holes, then "measles" (light areas in the film). It } is not worth ruining sections because one has used old Formvar solution. } Buy or make new from powder. Make a few films and check in the TEM for } stability, thickness, etc., before making a thousand. } Bye, } Hildy } I agree with Hildy. Test it first. "the proof is in the pudding". We store formvar for up to three years so far. Normally we make small amounts an use it till it is finished. We add Molecular sieve to keep it dry and store at ~4 Deg. C to reduce evaporation.
Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
In his request Rejane requested help with a particular=20 replication method and I provided this. I know this method=20 works well with at least some leaves - I've done it. The hairs on these particular leaves could be a problem.=20 Possibly the best solution is to pull the hairs off with a=20 first replica and then use for microscopy a second replica,=20 which is made on the same part of the leaf. Perhaps Rejane could obtain his stated goals: identifying=20 cell patterns and vein structures, by simply using a=20 dissecting scope. Anyway, the thin plastic replica would=20 give the option to use a low power compound light=20 microscope or to use SEM. Another correspondent (also a vendor) recommended the=20 alternative of his "special" silicone double replicating=20 kit. This material may have some uses, in this case it=20 would for instance make the process more complicated and=20 more expensive. He suggested: However, the use of acetone might not be desirable since it=20 is a solvent for the oils present on the leaf surface. Also, the dried=20 cellulose acetate film tends to be stiff and brittle and tends to "pull off"=20 some of the fine features from the sample making the "impression" difficult=20 to interpret. It is also an "inverted" or "negative" image of the surface,=20 something sometimes not so easy to interpret. The questioner expressed no interest in any oils, wax does=20 not readily dissolve in acetone (though it is probably=20 desirable to dissolve any waxes to see the features of=20 interest). I have not found such replicas stiff and it would be=20 desirable to remove the hairs to expose features. If the hairs were of interest, surely routine SEM of the=20 leaf should be the recommendation. The silicone replica is of no use under a compound=20 microscope. True the replica would be a negative, but why make a=20 positive for SEM (for TEM its another story)? SEMs have a polarity switch and a flick of that switch=20 turns a negative image of a single stage replica into a=20 positive image. A non-digital photograph, taken with a light microscope=20 could be reversed by making a contact print onto a bit of=20 TEM 4489 film to achieve a negative (therefore a positive=20 of the replica) on the paper print.
Some questioners are inexperienced and are not in a=20 position to properly evaluate given advice. I prefer to recommend a product only if convinced that this=20 would aid the work of the inquirer. As a vendor I may be=20 keen to make certain products known, but that should be at=20 my expense, eg advertising. It is not fair to use the=20 Listserver feature 'MY' products, unless its quite relevant=20 to the inquiry. Its abominable, to recommend an unsuitable=20 product when in the process an inexperienced person maybe=20 misled into using an unsuitable technique and a more=20 expensive product. True, we all have an ability to=20 'convince' ourselves that our actions are 'right', but I=20 think that most people can see the difference between=20 rationalising and self-delusion. Cheers Jim Darley ProSciTech Microscopy=20 PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au=20 *****
Rejane- Replicating with various plastic films is easy. Apply a drop or two of the recommended solvent (acetone or methyl acetate) onto the surface. Cover with a small rectangle of replicating film, this should have a notch near a corner to identify "specimen side". Apply for a moment gentle pressure, perhaps using a microscope slide. Leave to dry for five minutes. Pull the replica off. Angle coating (say 30 degrees) with metal helps to visualise depths for light microscopy, whereas for SEM, sputter coating is preferable. For TEM a secondary replica of carbon and metal would be required, but that is another story.
The replica would probably remove the plant hairs and a subsequently applied replica may be preferred. Also, many leaf surfaces are covered with wax. This may be removed by flushing the leaf prior to replication with xylene. Rejane, if you require more details email me direct. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Thursday, 3 September 1998 20:01, Rejane Magalh=E3es Pimentel Galindo [SMTP:ggalindo-at-elogica.com.br] wrote: } } I work with Atriplex nummularia and I want to examine } epidermal cell } patterns. This specie has a proffusion of glandular } hairs in both surfaces. } } I read in a paper something about use cellulose acetate } } film with acetone } } to make impressions of the leaf. } } } } Someone know something different and simplest about this } } subject? } } I also need to know what to do to take off these } } trichomes. } } Aside this, I want to obtain the vein impressions; I need } a simple method } to highlight the leaf venation. } } Thank in advance.
26th SMG Symposium at the West Park Conference centre, Univerity Dundee. Wednesday 11 November 98
1998 Final Programme 'Apoptosis' - when discretion is better than valour David Harrison, Dept of Pathology, University of Edinburgh.
'En Bloc' optical staining of resin embedde specimens using a confocal laser scanning microscope Ian Roberts, Scottish Crop Research Institute, Dundee
Cathodoluminescence microscopy Adrian Finch, Dept Environmental Sciences, University of Hertfordshire.
Blackcurrant fruit development in three dimensions by NMR microscopy and complimentary techniques Shelia Glidewell, Scottish Crop Research Institute, Dundee
Cryo Techniques and high resolution conventional SEM Alan Robbins, Oxford Instruments, Oxford
Quantitative immunoelectron microscopy - insights into mitotic membrane dynamics John Lucocq, Department Anatomy and Physiology, University of Dundee.
Microscopy in studies of plant surface, structure and function Chris Jeffree, Dept of Botany, University of Edinburgh.
Visit our Web Site at: http://www.abdn.ac.uk/~nhi691/smg98.htm
EM Unit, Dept Zoology University Of Aberdeen Tillydrone Avenue Aberdeen AB24 2TZ Tel 01224-272847 Fax 01224-272396
recently there was a discussion about scanners. If I'm remember well the Leaf scanner is now produce by a compagny named Brenson Inc. . Do some of you now these compagny and have they a sale representative in Europe ?
Thank's a lot
Marc
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
Here's a good trick. I recently wanted to do leaf surface impressions for stomatal counts and was experimenting with cellulose acetate etc. A physiologist friend comes along and says "have you tried ignition sealer?" Turns out this works really well. I sprayed the leaf surface with Kraco ignition sealant (this stuff is used as a spray waterproofing treatment for electrical wires and engine components), let it set for 5 min. and then peeled off the surface coating with clear tape. The replica/tape was then simply stuck to a glass slide and examined by phase contrast. This works well for epidermal cell shape, stomatal shape and distribution, and vein shape. I don't know how well it will work on trichomes and it probably depends on how elaborated they are. A word of caution; it seems that not all ignition sealer sprays are created equal. 'Wire Dryer' brand did not work at all. Other companies that manufacture this stuff include Hydrosol, Kleenflo and Spray-pak.
} } } =============================================== } } I work with Atriplex nummularia and I want to examine epidermal cell } } patterns. This specie has a proffusion of glandular hairs in both } } surfaces. I read in a paper something about use cellulose acetate film } } with acetone to make impressions of the leaf. } } How about gelatin? } } Jim Harper
________________________ C. John Runions, Ph.D. Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
Hello all, I've recently had a request to do some TEM of some PbSn solder bumps (on silicon ICs). I have some concerns about this, since the solder can begin to melt at about 150C. Has anyone been sucessful in making a TEM section of such a structure? I would be very interested in any tips or tricks.
Also, our local supplier of glassine envelopes for TEM negatives (3 3/4 x 2 3/4 inch) has told us they aren't stocking any more. Does anyone know of a UK-based supplier?
Many thanks in advance
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK
Hi all, in my earlier email about spraying ignition sealer onto leaves to make replicas I said that the product used was manufactured by Kraco. I lied. It's actually made by Krylon of Columbus Ohio, 43215. Cheers, John
________________________ C. John Runions, Ph.D. Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
} I've recently had a request to do some TEM of some PbSn solder bumps } (on silicon ICs). I have some concerns about this, since the solder can begin } to melt at about 150C. Has anyone been sucessful in making a TEM section of } such a structure? I would be very interested in any tips or tricks. } I suggest examining the specimen at ~-150 C. You do not need to cool the specimen prior to insertion in the TEM; just place the RT grid in the cryo holder, insert the holder into the TEM, then cool. Low dose rate will also help keep beam heating from producing locally high temperatures. Good luck. Yours, Bill Tivol
I am in the market for a digital camera that can fulfill a wide range of objectives, including fluorescence imaging of fura. The experiments would probably be time-varying, which would limit exposures to ~1/2 second at best. Judging by the fact that the systems I have seen use intensified CCD (ICCD) cameras, my impression is that fura produces a very weak signal. Does anyone know if there are any non-intensified cameras (eg back-illuminated) capable of the same performance, or are ICCD cameras the only way to go?
Thanks
Eric
Eric Johnston Department of Bioengineering University of Pennsylvania 120 Hayden Hall 3320 Smith Walk Philadelphia, PA 19104-6392 215-898-1958 (F) 215-573-2071 ericdj-at-seas.upenn.edu
I'm new at operating an SEM that has a tungsten filament. For the past 5 years I had an instrument that used a LaB6, but I've changed companies. I have 2 questions. 1. Both the people here and the instrument's service man tell me that the tungsten filament is more stable than the LaB6. I ask them to explain further and they really don't get into it. What is the stable thing about the tungsten filament? My LaB6 seemed fine to me, perfect in fact, so what is the unstableness of it? 2. I'm also told here that, with the tungsten filament, as the beam sits in one area on the sample, Carbon will develop in that area. Is this true? If so how long does it take for the carbon to contaminate the area, and also does this take any confidence in a carbon analysis and throw it out the window?
For a brief explanation check J.W. Edington. "Practical Electron Microscopy in Materials Science"
or you can check Hirsch et.al. "Electron Microscopy of Thin Crystals" Chapter 16.
Very briefly, Lorentz Microscopy is a technique used to image magnetic domains in ferromagnetic materials using a TEM. The name comes from the Lorentz formula ( i.e. v xB see an introductory Physics book) for the force on a charged particle moving in a magnetic field B.
Jordi Marti ---------- } From: "Pettieswa-at-aol.com"-at-sparc5.microscopy.com To: Microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
In a message dated 98-09-08 09:24:58 EDT, ericdj-at-seas.upenn.edu writes:
{ { I am in the market for a digital camera that can fulfill a wide range of objectives, including fluorescence imaging of fura. The experiments would probably be time- varying, which would limit exposures to ~1/2 second at best. Judging by the fact that the systems I have seen use intensified CCD (ICCD) cameras, my impression is that fura produces a very weak signal. Does anyone know if there are any non-intensified cameras (eg back- illuminated) capable of the same performance, or are ICCD cameras the only way to go? } }
Eric,
I'm not sure what a "back-illuminated non-intensified" camera is, but I would highly recommend a cooled CCD camera for your applications, probably something like the Spot or maybe the Spot Jr., from Diagnostic Instruments ($5-8K).
It depends on your application, but a 1/2 second exposure would be pushing it for just about any CCD camera if you want color. There are other high-end, high-speed digital cameras (maybe something from Dage MTI) that could do this in B&W, but they are in the range of $20-25K.
Hope this is of some help.
Bob ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 / (520) 546-4986 rchiovetti-at-aol.com
********************************* Cryostats / Microtomes / Tissue Processors / Embedding Centers / Slide Stainers / Glass Coverslippers / Microscopes (Representing Leica since 1967) / Fiber Optic Systems / Linear Measuring / Micromanipulation (Linear Encoded, Video) / Image Analysis, Archiving, Capture / Video (Cooled CCD, Digital, RGB, Super VHS, 3-chip) / Video Printers / Vibration Isolation Systems / Programmable Stages / Heating & Cooling Stages / Motion Control and Positioning Systems
At 10:37 AM 9/8/98 -0400, Mark Darus wrote: } } I'm new at operating an SEM that has a tungsten filament. For the past 5 } years I had an instrument that used a LaB6, but I've changed companies. } I have 2 questions. } 1. Both the people here and the instrument's service man tell me } that the tungsten filament is more stable than the LaB6. I ask them to } explain further and they really don't get into it. What is the stable thing } about the tungsten filament? My LaB6 seemed fine to me, perfect in fact, } so what is the unstableness of it?
I have only ever run tungsten, but the current has been stable for me. I can't say anything about instabilities. I have heard about field emission scopes sometimes being unstable, but even that has remedies.
} 2. I'm also told here that, with the tungsten filament, as the beam sits in } one area on the sample, Carbon will develop in that area. Is this true? If } so how long does it take for the carbon to contaminate the area, and also } does this take any confidence in a carbon analysis and throw it out the } window?
Last I heard, there was no difference between electrons once they left the gun, be it W, LaB6, or FE. The only difference would be in the number of them per time in a given space. The lesser vacuum requirements for a W filament might lead to more rapid C buildup on the sample. But our 840 with W normally runs 10-6 torr and has only moderate problems with buildup.
I am leary of doing much with C analyses. The absorption factors are significant sources of uncertainty. I would stick to qualitative comparisons between points as much as possible. And make sure spectra are collected under similar conditions of time and current.
Dear Mark, } } 1. Both the people here and the instrument's service man tell me } that the tungsten filament is more stable than the LaB6. I ask them to } explain further and they really don't get into it. What is the stable thing } about the tungsten filament? My LaB6 seemed fine to me, perfect in fact, } so what is the unstableness of it?
Can you put a Faraday cage in the beam? If so, you can measure the stability for periods longer than the response time of the cage+electronics. High-frequency instabilities are harder to measure, but would show up as light or dark bands in the image for appropriate scan rates. I have no experience with SEM, but I don't think there is much difference in stabil- ity between LaB6 and W. Both are thermionic sources which should be heated by DC current. In this case the temperature--therefore the emission--should be constant.
} 2. I'm also told here that, with the tungsten filament, as the beam sits in } one area on the sample, Carbon will develop in that area. Is this true? If } so how long does it take for the carbon to contaminate the area, and also } does this take any confidence in a carbon analysis and throw it out the } window? } This will happen with any source of electrons, but the contamination rate will vary with such parameters as vacuum in the specimen area and the nature of the specimen. The appearance of these contaminant peaks (they look like little mountains) has been used to measure local specimen thickness and to identify the position where EDS was done (for single-point analysis). The HVEM does not leave such peaks in spite of the so-so column vacuum and its use for plastic sections of biological specimens. Certainly, if you see carbon peaks on the specimen, you will also see carbon peaks in the EDS spec- tra, so I wouldn't trust a carbon analysis on such an instrument either for a single-point spectrum or for a carbon element map. Yours, Bill Tivol
I am looking for pointers to products or articles that address the following idea.
A single picture taken through a light microscope at say 100x has very small depth of focus.
But if a large number of pictures is taken at varying focal planes, the entire surface of a 3D subject can be imaged sharply, albeit one plane at a time.
It seems, in concept, that the sharply focused portions of each picture could be montaged together, to produce a single picture showing the entire 3D surface in sharp focus at once.
The required number of pictures could be quite large, but the montaging seems like a job that could be automated with computer image processing software.
I presume that this idea must have been studied somewhere. Perhaps there are products available to do the job. But I don't know them.
So I am asking for help. If you know of literature articles, products, or people who are working on this technique, then I would greatly appreciate getting a pointer to them.
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Dear Mark, } } 1. Both the people here and the instrument's service man tell me } that the tungsten filament is more stable than the LaB6. I ask them to } explain further and they really don't get into it. What is the stable thing } about the tungsten filament? My LaB6 seemed fine to me, perfect in fact, } so what is the unstableness of it?
Can you put a Faraday cage in the beam? If so, you can measure the stability for periods longer than the response time of the cage+electronics. High-frequency instabilities are harder to measure, but would show up as light or dark bands in the image for appropriate scan rates. I have no experience with SEM, but I don't think there is much difference in stabil- ity between LaB6 and W. Both are thermionic sources which should be heated by DC current. In this case the temperature--therefore the emission--should be constant.
} 2. I'm also told here that, with the tungsten filament, as the beam sits in } one area on the sample, Carbon will develop in that area. Is this true? If } so how long does it take for the carbon to contaminate the area, and also } does this take any confidence in a carbon analysis and throw it out the } window? } This will happen with any source of electrons, but the contamination rate will vary with such parameters as vacuum in the specimen area and the nature of the specimen. The appearance of these contaminant peaks (they look like little mountains) has been used to measure local specimen thickness and to identify the position where EDS was done (for single-point analysis). The HVEM does not leave such peaks in spite of the so-so column vacuum and its use for plastic sections of biological specimens. Certainly, if you see carbon peaks on the specimen, you will also see carbon peaks in the EDS spec- tra, so I wouldn't trust a carbon analysis on such an instrument either for a single-point spectrum or for a carbon element map. Yours, Bill Tivol
I would check with Diagnostic Instruments, Inc. Sterling Heights, Michigan 313-731-6000. There is probably a dealer in your area, but am not sure who.
Good Luck, C. Passione -----Original Message----- } From: Eric Johnston {ericdj-at-seas.upenn.edu} To: 'Microscopy' {Microscopy-at-sparc5.microscopy.com}
Thank you for all your responses to my need for a critical point dryer. I have purchased one, but have saved the information (vendors, models, prices, etc.) from the other offers I received. If anyone else is looking for a good price on a CPD, drop me a line, and I'll share the info.
The Joint Meeting of the Florida American Vacuum Society and the Florida Society for Microscsopy will be held the week following PITTCON (March 14-16, 1999) in Orlando on the campus of the University of Central Florida.
Events
golf tournament 2 day symposium (physical and biological sciences) including over 20 invited speakers student poster session and prizes: 1st prize: all expenses paid trip to attend national AVS or MSA meetings over 40 exhibitors AVS sponsored short courses equipment demos
AND Vendor Sponsored short courses:
1) Specimen Preparation using the Tripod Polisher (3/12-3/13): sponsored by South Bay Technology, instructor: Ron Anderson, IBM.
2) Specimen Preparation using Focused Ion Beam Milling (3/18-3/19): sponsored by FEI/Philips, instructors: Lucille Giannuzzi, UCF and Fred Stevie, Cirent Semiconductor.
Vendor space is limited and sells out early. Please contact Fred Stevie at stevie-at-lucent.com.
For other information, please contact Lucille Giannuzzi at lag-at-pegasus.cc.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-pegasus.cc.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
} } So I am asking for help. If you know of literature articles, } products, or people who are working on this technique, then I would } greatly appreciate getting a pointer to them. } } Please respond by email to } } Rik.Littlefield-at-pnl.gov (or rj_littlefield-at-pnl.gov) } } Thanks very much. } =================================================================== } Rik Littlefield } Senior Research Scientist } Pacific Northwest National Laboratory } Richland, WA 99352 } email: Rik.Littlefield-at-pnl.gov } phone: 509-375-3927 } fax: 509-375-3641
Hi,
Lots of information available for your interest, I'm deep into this kinda work. Since I work with 3-d reconstructions of injured spinal cord tissue and transplants and other things (you?) I'll generalize.
1. About sectioning as you mentioned, this can be done on thick sections via "optical sectioning" using a microscope with z-axis motor (Leica, for example), ours is computer driven with a CCD camera to automatically acquire a "stack" of images through thick tissue sections. However, depending on what you're considering, using physically sectioned (histological section) with embedded fiducial points for section registration can also be quite useful (lots of literature on that), thereafter, fairly large histological sections (say 25mm long by 15 mm wide) can be imaged under high resolution (less than 10 microns per pixel) which can allow reconstructions where optical section might not work (for large tissue regions). Course you won't get the resolution of 100x. IPLab by Scanalytics is a good program that might be helpful (forgot their URL).
2. You can take sections that are "in focus" based on distinct boundaries that can be automatically identified via algorithm. The freeware image program NIH-Image (http://rsb.info.nih.gov/nih-image/) with one of the myriad of already available macros (forgot the name of the specific one but you can do a gopher search on the NIH-Image website of the NIH-Image discussion list) can do this with a stack of images to create a composite (aside, I think you mean "composite" versus "montage" since we're dealing with 3-d and not strictly 2-d imaging). Oh yeah, if you are into imaging, get yourself on that NIH-Image discussion list. You'll meet other imaging experts there.
3. Also, there are programs such as Surfdriver (http://surfdriver.ml.org) which you can use to outline boundaries on a stack of images and create shaded surfaces of 3-d structures. Going deeper, you can check out VoxBlast (http://www.vaytek.com/) which does more sophisticated 3-D volume visualization and allows measurements and other interesting things (handles my 300-400 megabyte datasets like a charm). You don't mention digital deconvolution so we won't go there. Going deeper still, why go to all the trouble to reconstruct something in 3-D, without thinking spatial analysis? Perhaps, you want to know how close a dendrite surface is to a particular neuron and would like a 3-D color surface map (like a terrain map) detailing proximity (or it could be 3-D localized concentrations of transmitter or whatever). For that kind of custom work, check out IDL from RSI, Inc. (http://www.rsinc.com/) but now we're into programming. Point is, lots o' people want to visualize in 3-D, but the fun begins when you want to quantify in 3-D. Oh, and if you quantify, think "stereology."
For literature searches, I would suggest "3-D reconstruction" (text search) on Medline as a start and of course, check out the latest book (3rd edition) of John Russ's "The Image Processing Handbook" by CRC Press, and is just fabulous (unpaid endorsement) plus he has a way-cool course he teaches in North Carolina which I thought was great, plus he has way-cool software CD-ROM Image Processing Toolkit (http://members.aol.com/ImagProcTK/) with useful tutorials, very good. Not heavy into 3-D but the 2-D coverage is so useful you should just have it.
Well, enough gushing over 3-D and imaging, back to work for me, hope I said something useful. Brian C. Tryon MD/PhD student Allegheny University of the Health Sciences (Hey! We're bankrupt!) School of Medicine 3200 Henry Avenue Philadelphia, PA 19129 USA
I can speak from experience here since I have been using LaB6 & Tungsten for a number of years.
Tungsten in general will give a short lifetime ( typically 100 hrs+ ) whilst a LaB6 ( in theory ) will last for months. My experience with LaB6, which probably depends on the LaB6 supplier ( in this case Denka ), has produced approx. 6 weeks of more electrons before the filament starts to produce less and the alignment wander.
This may be due to the type of source of the LaB6, but I have talked recently to someone running a different supplier's LaB6 and he was suffering the same problems.
In general operation the LaB6 will not give noticable differences in stability. Any measurements using a Faraday cage & monitor will give different results depending on the age of the filament. However if you have demanding customers, such as Geologists ( apologies to any reading this ! ), when you are carrying out quant analyses and producing totals approaching 100% the analyses are very dependant on beam stability. I have not been able to carry out these analyses without using a tungsten source. Even with a tungsten filament it will be unstable in the early & late part of its life and serious analyses can only be carried out during the middle period. I produce longer life and greater stability by cleaning the gun properly after a blown filament and using the Ion pump to produce a higher vacuum during tungsten operation. Tungsten filaments also have faster ramp times and are more tolerant of poor vacuums.
As regards carbon build up I think that it is both vacuum & sample dependant. With more hydrocarbons in the vacuum you get a greater build-up. I have two turbo-molecular pumped systems and do not suffer from much carbon build-up unless certain samples are scanned.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Mark Darus {DARUSM-at-cle.lg.bfg.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Although I am a "vendor", allow me to comment on this thread, from the perspective of someone who has manufactured tungsten filaments and distributed Denka LaB6 cathodes for more than 10 years.
Colin Reid wrote:
} Tungsten in general will give a short lifetime ( typically 100 hrs+ ) whilst } a LaB6 ( in theory ) will last for months. My experience with LaB6, which } probably depends on the LaB6 supplier ( in this case Denka ), has produced } approx. 6 weeks of more electrons before the filament starts to produce less } and the alignment wander. This may be due to the type of source of the LaB6, but I have } talked recently to someone running a different supplier's LaB6 and he was suffering } the same problems.
For any filament, by far the most important parameter affecting material loss, and, therefore, lifetime is vacuum. This holds true for both tungsten filaments and LaB6. Filament life, therefore, is highly dependent on the vacuum conditions of a given microscope, and filament life will vary substantially from microscope to microscope.
Tungsten filaments are cold-formed from wire, and the stress from the forming will cause instability if the filaments are not properly annealed under vacuum. If the tungsten filament has not been properly annealed by the manufacturer, the user will, in effect, anneal it in the microscope. The filament will be unstable during this process, and will often have to be realigned after the annealing.
In the case of LaB6, the crystal is in {100} orientation, and formed into a point, a round tip or a microflat tip. As the cathode experiences material loss, the tip flattens. After some 150 hours, the brightness will fall off, as the emission surface of the tip forms into a larger flat. In effect, the sharp or round tips become flat tips. Although the cathode has hundreds of hours of "life" left, it will never again be as bright as it was during the first ~150 hours. This is true of any manufacturer's LaB6.
For quantitative analysis, mechanical stability becomes an issue. For this application, the LaB6 mount should be as robust as possible.
I would be happy to provide additional details to anyone interested. In particular, we have several Denka technical reports which explain the relationship between lifetime, brightness, vacuum and material loss for LaB6 cathodes.
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
I saw a demo in London of a software package "Auto-Montage" from Synoptics which was combining 5 and more partially focussed images to one fully focused one, using a frame-grabber. E-mail address : sales-at-synoptics.co.uk With best regards, emond de Roever
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I am looking for pointers to products or articles that address the following idea.
A single picture taken through a light microscope at say 100x has very small depth of focus.
But if a large number of pictures is taken at varying focal planes, the entire surface of a 3D subject can be imaged sharply, albeit one plane at a time.
It seems, in concept, that the sharply focused portions of each picture could be montaged together, to produce a single picture showing the entire 3D surface in sharp focus at once.
The required number of pictures could be quite large, but the montaging seems like a job that could be automated with computer image processing software.
I presume that this idea must have been studied somewhere. Perhaps there are products available to do the job. But I don't know them.
So I am asking for help. If you know of literature articles, products, or people who are working on this technique, then I would greatly appreciate getting a pointer to them.
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Hi Mark, } } 1. Both the people here and the instrument's service man tell me } that the tungsten filament is more stable than the LaB6. I ask them to } explain further and they really don't get into it. What is the stable thing } about the tungsten filament? My LaB6 seemed fine to me, perfect in fact, } so what is the unstableness of it?
I have experiences on both tungsten and field emission SEMs. The emission of cold FE gun does have fluctuation. But at normal condition, the fluctuation is tolerable and can't be seen on the final images.
} 2. I'm also told here that, with the tungsten filament, as the beam sits in } one area on the sample, Carbon will develop in that area. Is this true? If } so how long does it take for the carbon to contaminate the area, and also } does this take any confidence in a carbon analysis and throw it out the } window? }
The carbon build-up, mostly hydrocarbon, is depended on vacuum and sample. Especially the "cleanliness" of the vacuum. I have worked on two different FESEMs, both are equipped with turbo pumps. The difference is that one scope is backed by a rotary pump, the other is a totally oil-free pumping system. I looked the same test sample using both scopes and found that there is a big difference on the contamination rate. I can multiple-scan the same area at the magnification of 500,000x using the FESEM equipped with totally oil-free pumping system, but not the other one.
Ya Chen
======================================================================== \ / Integrated Microscopy Resource (IMR)-- \ / __ a NIH Biomedical Research Resource TEL: 608-263-8481 \/ / / University of Wisconsin-Madison FAX: 608-265-4076 / / / 1675 Observatory Drive #159 / /__/_ Madison, WI 53706 Email: ychen14-at-facstaff.wisc.edu ======================================================================== IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr/home.htm
You have accurately described a technique which is solved using several different approaches.
The most common commercialized version is confocal microscopy. The best general reference is the {underline} Handbook of Biological Confocal Microscopy {/underline} , published by Plenum and edited by Dr. James Pawley. We also have a short discussion in our book, {underline} Optimizing Light Microscopy for Biological and Clinical Laboratories {/underline} (see our web site: { {http://www.MME-Microscopy.com/education} ). You may also want to visit the web sites of the confocal manufacturers. The best metasites for locating those URLs are either the MSA manufacturers' list or MicroWorld resources and news (www.mwrn.com).
A scond commercialized approach is the 3-D imaging offered through Edge Scientific. I believe their website is listed on both the MSA and mwrn sites. They provide a variety of options, ranging from a full blown system (the R400), to a retrofittable reflected light/fluorescence head and the new "Perceptra", a retrofittable, 3-D illuminator.
I have also heard of non-commercial work done by people such as Dr. Shinya Inoue (retired from the Woods Hole Biological Institute) who have used DIC to take very thin optical sections then used computer programs to reconstruct the resulting set of serial optical sections.
There is a company that does exactly what you are describing. The company is called Soft Imaging system Corporation. I believe, they are in Denver. I know they have a website, but I can't remember the URL right now. I'll try to find out and send it to you. Perhaps somebody else knows it.
Gloria
} } } ---------- } From: Rik Littlefield[SMTP:rj_littlefield-at-pnl.gov] } Sent: Tuesday, September 8, 1998 1:02 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Photomontaging for increased depth of field ? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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There is a company that does exactly what you are describing. The company is called Soft Imaging system Corporation. I believe, they are in Denver. I know they have a website, but I can't remember the URL right now. I'll try to find out and send it to you. Perhaps somebody else knows it.
Gloria
} } } ---------- } From: Rik Littlefield[SMTP:rj_littlefield-at-pnl.gov] } Sent: Tuesday, September 8, 1998 1:02 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Photomontaging for increased depth of field ? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi Mark, } } 1. Both the people here and the instrument's service man tell me } that the tungsten filament is more stable than the LaB6. I ask them to } explain further and they really don't get into it. What is the stable thing } about the tungsten filament? My LaB6 seemed fine to me, perfect in fact, } so what is the unstableness of it?
I have experiences on both tungsten and field emission SEMs. The emission of cold FE gun does have fluctuation. But at normal condition, the fluctuation is tolerable and can't be seen on the final images.
} 2. I'm also told here that, with the tungsten filament, as the beam sits in } one area on the sample, Carbon will develop in that area. Is this true? If } so how long does it take for the carbon to contaminate the area, and also } does this take any confidence in a carbon analysis and throw it out the } window? }
The carbon build-up, mostly hydrocarbon, is depended on vacuum and sample. Especially the "cleanliness" of the vacuum. I have worked on two different FESEMs, both are equipped with turbo pumps. The difference is that one scope is backed by a rotary pump, the other is a totally oil-free pumping system. I looked the same test sample using both scopes and found that there is a big difference on the contamination rate. I can multiple-scan the same area at the magnification of 500,000x using the FESEM equipped with totally oil-free pumping system, but not the other one.
Ya Chen
======================================================================== \ / Integrated Microscopy Resource (IMR)-- \ / __ a NIH Biomedical Research Resource TEL: 608-263-8481 \/ / / University of Wisconsin-Madison FAX: 608-265-4076 / / / 1675 Observatory Drive #159 / /__/_ Madison, WI 53706 Email: ychen14-at-facstaff.wisc.edu ======================================================================== IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr/home.htm
Gut tissue fixation I am looking for a protocol for intestinal tissue. I have opossum intestinal tissue fixed by a colleague that has a lot of OsO4 pepper. I have also received gut tissue that appeared to have undergone autolysis. I am expecting more of these samples and would like to recommend a change in protocol. It is probably not possible to use perfusion. I am looking for intercellular and intracellular bacteria and /or protozoal infections in these tissues.=20 Would the chemistry of the intestine be causing this type of problem? Or is it simply a matter of inadequate washing?=20 Thanks=85=85.. Sally
The fixation used was:=20 2% glut in PBS at pH 7.2 for 3 hours. 3X washed in PBS 1X washed in H20 1% OsO4 for 4H room T washed 3x in H20 dehydrated in a 25% series of acetone Infiltrated and embedded in a 25% series into a Spurrs/ Quetol blend.
Sally Burns Center for Electron Optics B5 Pesticide Research Center (517) 355-5004
We have a Hitachi S-570 SEM and we would like to take "motion pictures" of MEMS devices inside the chamber using the SEM. It does have a "video out" connector on its pc board, however, the signal from it is not a standard NTSC video signal. I've talked to Hitachi and was told that at one time they made a coverter, but that it is no longer available. Does anyone, by any chance, know if the video signal conforms to some other video standard? Also, does anyone know of a source for either the Hitachi converters or some other converter that would give us a usable video signal to feed into monitor or video recorder?
Janet Rice MCC Senior Member Technical Staff rice-at-mcc.com 512-338-3266
Hi Mark, } } 1. Both the people here and the instrument's service man tell me } that the tungsten filament is more stable than the LaB6. I ask them to } explain further and they really don't get into it. What is the stable thing } about the tungsten filament? My LaB6 seemed fine to me, perfect in fact, } so what is the unstableness of it?
I have experiences on both tungsten and field emission SEMs. The emission of cold FE gun does have fluctuation. But at normal condition, the fluctuation is tolerable and can't be seen on the final images.
} 2. I'm also told here that, with the tungsten filament, as the beam sits in } one area on the sample, Carbon will develop in that area. Is this true? If } so how long does it take for the carbon to contaminate the area, and also } does this take any confidence in a carbon analysis and throw it out the } window? }
The carbon build-up, mostly hydrocarbon, is depended on vacuum and sample. Especially the "cleanliness" of the vacuum. I have worked on two different FESEMs, both are equipped with turbo pumps. The difference is that one scope is backed by a rotary pump, the other is a totally oil-free pumping system. I looked the same test sample using both scopes and found that there is a big difference on the contamination rate. I can multiple-scan the same area at the magnification of 500,000x using the FESEM equipped with totally oil-free pumping system, but not the other one.
Ya Chen
======================================================================== \ / Integrated Microscopy Resource (IMR)-- \ / __ a NIH Biomedical Research Resource TEL: 608-263-8481 \/ / / University of Wisconsin-Madison FAX: 608-265-4076 / / / 1675 Observatory Drive #159 / /__/_ Madison, WI 53706 Email: ychen14-at-facstaff.wisc.edu ======================================================================== IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr/home.htm
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Date 9/9/98 Time: 3:54 PM Internal Memorandum
We currently have two positions open for SEM/EDS materials technicians at Exponent Failure Analysis Associates in Menlo Park, CA. See attachment for details and send resume/information to hr-at-exponent.com.
by arwen.otago.ac.nz (8.9.1/8.9.1) with ESMTP id NAA17599; Thu, 10 Sep 1998 13:21:44 +1200 (NZST) X-Sender: st004716-at-brandywine.otago.ac.nz Message-Id: {l03130302b21cdafa0cf4-at-[139.80.34.56]} In-Reply-To: {3.0.32.19980909161244.0090e6d0-at-pilot.msu.edu} Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
} Gut tissue fixation } I am looking for a protocol for intestinal tissue. I have opossum } intestinal tissue fixed by a colleague that has a lot of OsO4 pepper. I } have also received gut tissue that appeared to have undergone autolysis. I } am expecting more of these samples and would like to recommend a change in } protocol. It is probably not possible to use perfusion. } I am looking for intercellular and intracellular bacteria and /or } protozoal infections in these tissues. } Would the chemistry of the intestine be causing this type of } problem? Or } is it simply a matter of inadequate washing? } Thanks=85=85.. Sally } } The fixation used was: } 2% glut in PBS at pH 7.2 for 3 hours. } 3X washed in PBS } 1X washed in H20 } 1% OsO4 for 4H room T } washed 3x in H20 } dehydrated in a 25% series of acetone } Infiltrated and embedded in a 25% series into a Spurrs/ Quetol blend.
Sally, Are you sure that is OsO4 pepper? Could it be PO4 pepper? Although it seems that there are heaps of water washes in your protocol anyway, so possibly may not be. We have processed this stuff before, using perfusion admitedly, but using Cacodylate buffer, havent had too many probs as far as I can remember. I would change buffer initially to have a go. Is it necessary to fix for 4hrs? We routinely fix in 1%OsO4 in buffer for 1hr at RT. Perhaps having it in here for too long would cause excess OsO4 deposition. About the apparent autolysis, could it be that there is lots of mucus/garbage around the tissue preventing fix from getting in? Would a paraformaldehyde/glut fix be better?
Just thinking aloud.............! Hope you find the cure to your problems,
Rich.
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
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=3EGut tissue fixation =3E I am looking for a protocol for intestinal tissue=2E I have = opossum =3Eintestinal tissue fixed by a colleague that has a lot of OsO4 pepper= =2E I =3Ehave also received gut tissue that appeared to have undergone autoly= sis=2E I =3Eam expecting more of these samples and would like to recommend a cha= nge in =3Eprotocol=2E It is probably not possible to use perfusion=2E =3E I am looking for intercellular and intracellular bacteria an= d /or =3Eprotozoal infections in these tissues=2E =3E Would the chemistry of the intestine be causing this type of=
=3Eproblem=3F Or =3Eis it simply a matter of inadequate washing=3F =3EThanks=85=85=2E=2E Sally =3E =3EThe fixation used was=3A =3E2% glut in PBS at pH 7=2E2 for 3 hours=2E =3E3X washed in PBS =3E1X washed in H20 =3E1% OsO4 for 4H room T =3Ewashed 3x in H20 =3Edehydrated in a 25% series of acetone =3EInfiltrated and embedded in a 25% series into a Spurrs=2F Quetol ble= nd=2E
Sally=2C Are you sure that is OsO4 pepper=3F Could it be PO4 pepper=3F Althoug= h it seems that there are heaps of water washes in your protocol anyway=2C s= o possibly may not be=2E We have processed this stuff before=2C using pe= rfusion admitedly=2C but using Cacodylate buffer=2C havent had too many probs a= s far as I can remember=2E I would change buffer initially to have a go=2E Is it necessary to fix for 4hrs=3F We routinely fix in 1%OsO4 in buffe= r for 1hr at RT=2E Perhaps having it in here for too long would cause excess= OsO4 deposition=2E About the apparent autolysis=2C could it be that there is lots of mucus=2Fgarbage around the tissue preventing fix from getting in=3F Wo= uld a paraformaldehyde=2Fglut fix be better=3F
Just thinking aloud=2E=2E=2E=2E=2E=2E=2E=2E=2E=2E=2E=2E=2E! Hope you find the cure to your problems=2C
Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P=2EO=2E Box 913 Dunedin New Zealand=2E Tel=2E National 03 479 7301 Fax=2E National 03 479 7254
=22Southernmost EM Unit in the World!=22 -----------------------------------------------------------------------= -
We are selling our 5 year old Leo(Leica) S440, with availibility approx. = 2-3 months. If anyone ( Ireland & possibly UK ) is interested will you = contact me directly for full details.
Thanks,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
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=3EGut tissue fixation =3E I am looking for a protocol for intestinal tissue=2E I have = opossum =3Eintestinal tissue fixed by a colleague that has a lot of OsO4 pepper= =2E I =3Ehave also received gut tissue that appeared to have undergone autoly= sis=2E I =3Eam expecting more of these samples and would like to recommend a cha= nge in =3Eprotocol=2E It is probably not possible to use perfusion=2E =3E I am looking for intercellular and intracellular bacteria an= d /or =3Eprotozoal infections in these tissues=2E =3E Would the chemistry of the intestine be causing this type of=
=3Eproblem=3F Or =3Eis it simply a matter of inadequate washing=3F =3EThanks=85=85=2E=2E Sally =3E =3EThe fixation used was=3A =3E2% glut in PBS at pH 7=2E2 for 3 hours=2E =3E3X washed in PBS =3E1X washed in H20 =3E1% OsO4 for 4H room T =3Ewashed 3x in H20 =3Edehydrated in a 25% series of acetone =3EInfiltrated and embedded in a 25% series into a Spurrs=2F Quetol ble= nd=2E
Sally=2C Are you sure that is OsO4 pepper=3F Could it be PO4 pepper=3F Althoug= h it seems that there are heaps of water washes in your protocol anyway=2C s= o possibly may not be=2E We have processed this stuff before=2C using pe= rfusion admitedly=2C but using Cacodylate buffer=2C havent had too many probs a= s far as I can remember=2E I would change buffer initially to have a go=2E Is it necessary to fix for 4hrs=3F We routinely fix in 1%OsO4 in buffe= r for 1hr at RT=2E Perhaps having it in here for too long would cause excess= OsO4 deposition=2E About the apparent autolysis=2C could it be that there is lots of mucus=2Fgarbage around the tissue preventing fix from getting in=3F Wo= uld a paraformaldehyde=2Fglut fix be better=3F
Just thinking aloud=2E=2E=2E=2E=2E=2E=2E=2E=2E=2E=2E=2E=2E! Hope you find the cure to your problems=2C
Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P=2EO=2E Box 913 Dunedin New Zealand=2E Tel=2E National 03 479 7301 Fax=2E National 03 479 7254
=22Southernmost EM Unit in the World!=22 -----------------------------------------------------------------------= -
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Fully operational JEOL JEM 100B with a W filament available in Export, = PA. Also includes numerous spare components. ALL offers considered, = best offer accepted.
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Hello all, Has anyone experienced puckering of tissues embedded in LRWhite resin? My = tissues are cell cultures fixed in 2%PFA, .5%GA, dehydrated to75% ETOH = then into graded LRWhite resin. I cut on a diamond knife and embed on 200 = mesh grids. Before staining, the puckers and wrinkles are seen as small, = and frequen, folds over most cells or along the cell borders. The resin = areas are totally free of folding. It's as if the tissue area is picking = up H2O during sectioning then has no where to go when it re-dries....very = frustrating. After staining it's worse, as the stain seems to stay in the = folds and gets really dark. Any thoughts and suggestions are very welcome = as always. Linda Fox Loyola University=20 Stritch School of Medicine lfox1-at-wpo.it.luc.edu
So called "osmium pepper" may be caused by the following:
1. Incompletely depolymerized paraformaldehyde or old glutaraldehyde that has started to polymerize. The aldehyde "globs" are left behind in the tissue to then react with the osmium and form a very fine precipitate. Do you see the pepper only in tissue? If so, this is a possibility.
2. Incomplete removal of PO4, as described by Richard Lander.
3. Lead stain that is precipitating (just starting to go bad). Is the pepper everywhere (even where tissue is absent)? If so, this is a possibility. Do unstained sections show the pepper? If not, then stain can be ruled out.
Best solutions: use freshly prepared, EM-grade aldehydes, cacodylate buffer, verified hi quality lead stain.
} } I am looking for a protocol for intestinal tissue. I have opossum } } intestinal tissue fixed by a colleague that has a lot of OsO4 pepper. I } } have also received gut tissue that appeared to have undergone autolysis. I } } am expecting more of these samples and would like to recommend a change in } } protocol. It is probably not possible to use perfusion.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
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Manhattan, New York City:
If I could be helpful to your lab or imaging department, I may accept payment in exchange for education: as an intern, or barter, or as a volunteer "tryout". (Salary when appropriate.)
-------------
-Currently I work as a commercial web and print designer and use Photoshop imaging software daily for the past 4 years. -I have been freelancing; coloring microscans in photohop for a stock photography company. -Also, well practiced in nature photography -I have designed web pages* for the invertebrate scientists at the American Museum of Natural History and examined the light micoscope in the histology lab. I have also observed the Interdepartmental Labs' SEM and Confocal. As a layman, I find the visual aspect of scientific analysis is fascinating. *http://research.amnh.org/invertebrates *http://www.renorex.com/idltest (under construction)
Art and Science: I would like to continue combining my visual imaging skills with my general interest in Science. How can I learn microscopy equipment and/or 3D animated software and thereby provide assistance to scientists.
Thank you in advance for any education or vocation advice you have time to give.
Renee Recker 16 W. 16th St. NYC, NY 10011 212-675-1665
http://www.renorex.com (portions require shockwave and Netscape 4.0 browser)
P.S.
Previously I have posted e-mail asking about how to obtain microscopy education in a college program. I am still amazed and cannot thank enough the generous personal and professional responses. But, still stuck at the starting gate. The only course in commuting distance is NYU Spring '99. So, I thought I might ask about on-the-job training.
A few years ago I prepared some intestinal tissue for use in a probiotics study. Although my main aim was the fixation of the mucus blanket, it was also necessary to fix the intestine itself.
The protocol I used was an unconventional one - an anhydrous fixation, where osmium in solvent was used as the primary fixative. No "pepper" was observed, but the images, of course, are different from ones obtained using a conventional protocol where glutaraldehyde in buffer was used as the primary fixative. This may be OK for you, depending on what you want to see.
The following reference gives the details of the protocol, and shows the SEM results. The equivalent TEM results, which are as yet unpublished, looked good, too.
Allan-Wojtas, P., Farnworth, E.R., Modler, H.W. and Carbyn, S. (1997). A solvent-based fixative for electron microscopy to improve retention and visualization of the intestinal mucus blanket for probiotics studies. Microscopy Research and Technique 36:390-399.
If you would like a reprint, please contact me offline with your address and other necessary information, and I send one to you.
Just another idea.
Good Luck.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
I have been asked to post the following question. One of our students wants to selectively stain polystyrene (to differentiate from polyethylene oxide) and has read that this has been done by ruthenium oxide vapour staining. Has anyone had any experience with this that they could share with us?
TIA,
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-medcor.mcgill.ca
Yes I see it. There is apparently a server at okstate.edu which may be the culprit. I have added them as well as the 2 addresses which are associated to the bounding mail
Rich Lander and Y Chen
to the filter. That should stop the mail from reaching the rest of you until we can figure out why it started.
Richard and Ya you'lll be in libo for awhile. You should be able to receive Email from Microscopy but not post messages.
Dear Listers, I will need to reorder Mo grids. The (very old) ones I've been using have very ragged grid bars--often with holes in them--and I'd like to find some better-quality ones. Has anyone found a vendor for really good Mo grids? Any vendor who thinks (s)he qualifies is welcome to email me directly. TIA. Yours, Bill Tivol
Project MICRO has been contacted by an energetic 4th grade teacher in Marin County, CA. She's been doing some microscopy and wants to do more, using the new MSA/LHS "Microscopic Explorations". She needs help now with cleaning her scopes, and could use a volunteer to help with the microscopy later. Do any of you folks live in Marin? Reply offlist, to me or direct to her (cc to me, please?)
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I would like to contact Dr. O.P. Ottersen, who works in the department of Anatomy at the University of Oslo, Norway. Does anyone have his e-mail address, per chance?
Thank you in advance,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
The needle valves (vent, fill and drain) on our Ladd critical point drier are quite diffucult to turn, even when warm. Does anyone know if they can be lubricated and, if so, how?
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
I have a method for an Oil Red O stain for GMA sections. It uses 60% aqueous triethyl phosphate and 5% aqueous ferric ammonium sulphate, however I can only locate these chemicals in the powder form. Does anyone know if it matters if the powder form is used and what they are likely to be dissolved in as it doesn't state this in the methodology?
Elizabeth Cox Fisheries Biologist Queensland Department of Primary Industries Northern Fisheries Centre, PO Box 5396, Cairns Qld Australia 4870
Does anayone on this network have experience with the new field of Raman Imaging ? We have a raman instrument with a tunable filter and imaging. I have a couple of questions. I am a TEM person and new at this so excuse me if these questions are too elementary.
The manufacturer claims that on aquiring the image the spatial resolution of the 'spectra' is as small as a pixel. But since unlike IR, Raman is a scattering process wouldn't all regions limited by the beam (probe) , about 150 microns, be affected by each other and thus negating the claim of 1 pixel resolution? Infact I did observe this experimentaly while imaging composite polymeric samples. Spectra from all regions illuminated by the beam are almost identical although the image does exhibit contrast due to different phases.
Thanks
Kalpana
******************************************************************************** **** Kalpana S Katti, Ph.D. Department of Polymers and Coatings North Dakota State University Fargo, ND 58105 ph:(701)231-8410 fax:(701)231-8439 email:kkatti-at-prairie.nodak.edu ******************************************************************************** ****
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } Has anyone experienced puckering of tissues embedded in LRWhite resin? My tissues are cell cultures fixed in 2%PFA, .5%GA, dehydrated to75% ETOH then into graded LRWhite resin. I cut on a diamond knife and embed on 200 mesh grids. Before staining, the puckers and wrinkles are seen as small, and frequen, folds over most cells or along the cell borders. The resin areas are totally free of folding. It's as if the tissue area is picking up H2O during sectioning then has no where to go when it re-dries....very frustrating. After staining it's worse, as the stain seems to stay in the folds and gets really dark. Any thoughts and suggestions are very welcome as always. } Linda Fox } Loyola University } Stritch School of Medicine } lfox1-at-wpo.it.luc.edu } } Hi, You have several problems here. First, acrylics like LR White, etc., do not bond WITH the tissue as do epoxies. This allows a lot of shifting as soon as the stresses are relieved by cutting a thin section. LR White also is modestly poorly crosslinked (as compared to the epoxides). You can increase the crosslinkage chemically (unless you are doing immunostaining, then you do not want to heavily crosslink). What to do? Do you need to use LR White? What is your purpose of using it. It has a number of downsides which one can trade off in the immunoprocessing protocols for better location of antigens. For standard TEM work is is inferior to epoxides (wrinkling of thick sections, poor crosslinkage, polymerization damage, difficulty sectioning because of its property of attracting water to the block face, etc.) What to do? If you must use LR White, try using a 100 mesh grid with a film. Also when picking up floating grids, DO NOT suck the water off under the grid with filter paper. Do draw the water off by sticking filter paper between the forceps. Then, lay the grid in the forcepts down to dry. Pull a lamp with a 60W bulb down over the forceps to slightly warm the situation. Use at least 5 forceps at once. That way some grids are drying and others are being picked up. This is the recommendation of Newman who holds the patent for LR White and LR Gold (He sold the license to the London Resin Company.) Should you decide to more heavily crosslink the resin, contact EMS for advice and the chemical. (I have no stock in EMS). If your sections have folds and you posstain them with heavy metals, the stain will accumulate in the folds. There is nothing you can do about that. You must avoid the wrinkles in the first place. Should you have more trouble, contact me. Bye, Hildy
Pat: Both polystyrene and Polyethylene oxide will stain with ruthenium oxide. However they may stain at different rates. There is a paper by Trent et al in Macromolecules 1983 v16 pg. 589-598 that descibes RuO4 staining of polymers. RuO4 is quite hazardous so be aware of the safety issues. Steve
At 01:12 PM 9/10/98 -0400, you wrote: } } I have been asked to post the following question. One of our students wants } to selectively stain polystyrene (to differentiate from polyethylene oxide) } and has read that this has been done by ruthenium oxide vapour staining. } Has anyone had any experience with this that they could share with us? } } TIA, } } Pat Hales } McGill University } Dept. of Anatomy & Cell Biology } hales-at-medcor.mcgill.ca } } }
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
John Mike Taylor of the Electrical Engineering Dept at the University of Delaware is in need of price and delivery of a 40 pin EPROM made by Intel or compatible No. 8755AD or D8755A. If anyone out there has any source please contact John Mike Taylor directly at jtaylor-at-ee.udel.edu. Thanks in advance Pete Dondl, P & S Products
Responding to the message of {v04003a09b21dc32600cf-at-[141.233.130.134]} from "wise-at-vaxa.cis.uwosh.edu"-at-Sparc5.Microscopy.Com: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } The needle valves (vent, fill and drain) on our Ladd critical point drier } are quite diffucult to turn, even when warm. Does anyone know if they can } be lubricated and, if so, how?
I have the same problem with my Ladd CPD. Other than that, I find the Ladd CPD to be a very good device (IHNCIIL,JASC). I allow users of the CPD to loosen the valves with pliers, but NOT to tighten them that way, just use finger pressure to tighten. But I suspect that those with delicate fingers cheat a little when I'm out of the room, but hopefully, without over-tightening the valves. So far so good. We've had the unit for over 12 years and never had to replace a valve.
Is it possible to adjust the turning tension on these needle valves??
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Michael T. Marshall Research Engineer, Electron Microscopy University of Illinois at Urbana-Champaign Frederick Seitz Materials Research Laboratory 104 South Goodwin avenue Urbana, IL 61801-2985 (217) 244-8193 fax: (217) 244-2278
Hi =20 Can anyone explain what the shape factor is (image analyses softwares) =20 As I understand it, when it is equal to 1 it means the object is a=20 disc...what if it is say 2.5 and 0.3?? =20 =20 Thanks =20 F.
wise-at-vaxa.cis.uwosh.edu-at-Sparc5.Microscopy.Com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } The needle valves (vent, fill and drain) on our Ladd critical point drier } are quite diffucult to turn, even when warm. Does anyone know if they can } be lubricated and, if so, how? } } Bob } } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } www.uwosh.edu/departments/biology/wise/wise.html
Dear Dr. Robert R. Wise,
RE: NEEDLE VALVES ON CPD
Try graphite or teflon grease. If this doesn't help than they may be wearing out and will probably need replacing if the problem gets too severe.
John Arnott --
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
} Does anayone on this network have experience with the new field of Raman } Imaging ? We have a raman instrument with a tunable filter and imaging. I } have a couple of questions. I am a TEM person and new at this so excuse } me if these questions are too elementary. } } The manufacturer claims that on aquiring the image the spatial resolution } of the 'spectra' is as small as a pixel. But since unlike IR, Raman is a } scattering process wouldn't all regions limited by the beam (probe) , } about 150 microns, be affected by each other and thus negating the claim } of 1 pixel resolution? Infact I did observe this experimentaly while } imaging composite polymeric samples. Spectra from all regions illuminated } by the beam are almost identical although the image } does exhibit contrast due to different phases.
I assume that you have an instrument wherein the sample is imaged onto a CCD, but in between there is a tunable filter so that at any one instant only a narrow range of wavelengths is striking the CCD. In this case light that is scattered from the sample will be imaged onto the CCD in the place corresponding to the place on the sample it came from.
I have been reading a lot of patents lately and it has made my exposition a little stilted, sorry.
Hope this helps. best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
Jack and Jill go to court. Jill takes the stand and lies her head off. Jack takes the stand and lies his head off. Who's telling the truth?
We have an opening for a post-doctoral research associate to work on a project involving in situ straining of fcc metals. The purpose is to study dislocation intersections and to compare experimental observations with large-scale atomistic simulations. Experimental techniques include dynamic imaging, weak-beam dark-field microscopy and high resolution electron microscopy. Experience with in-situ straining is highly desirable but not essential.
Please send resumes and the names of three references to the address below or attach to an e-mail reply. The post-doctoral program at Los Alamos is described at http://stb.lanl.gov/postdoc/postdoc.html
Terence E. Mitchell Laboratory Fellow Center for Materials Science MS-K765 Los Alamos, NM 87545 voice mail: 505-667-0938 fax: 505-665-2992 e-mail: temitchell-at-lanl.gov http://www.mst.lanl.gov/cms/welcome.html http://www.mst.lanl.gov/CMS/EMF/emf-people.html
I need to get in touch with a company that used to be called GW Electronics and they used to be in Norcross, Ga. My "new address and phone number" card is dated Sept. 1, 1983. Are they still around?
Bill -- ============================================================= Bill Chissoe III Electron Microscopist,University of Oklahoma E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
Shape factors can be really useful for morphometry studies but I've seen some poor formulas used that miss a lot of detail. You have to look at the particular formula to see what the investigator is trying to describe, hopefully, they would give you a few examples of the object of interest and its value so the reader can gauge the meaning and significance of different values.
A good stereology text (unpaid endorsement) that talks a bit about this that I would recommend is:
Russ, J. C. (1986). Practical Stereology. New York, Plenum Press.
I think these journal articles might be helpful too:
True, L. D. (1996). "Morphometric applications in anatomic pathology [Review]." Human Pathology 27(5): 450-467.
Royet, J. P. (1991). "Stereology: a method for analyzing images." Progress in Neurobiology 37(5): 433-74.
Prakash, Y. S., K. G. Smithson, et al. (1993). "Measurements of motorneuron somal volumes using laser confocal microscopy: Comparisons with shape-based stereological estimations." Neuroimage 1: 95-107.
Lastly, a great resource (like this discussion list) for image analysis questions is to go to the NIH-Image website at http://rsb.info.nih.gov/nih-image/, and do a Gopher search of the discussion list archive, which is a terrific resource (plus join the discussion list!). If you didn't know, NIH-Image is free image analysis software, and has a bunch of free macro code that can be easily modified to fit many image analysis needs, and it is still FREE! Yahoo!
good hunting,
Brian C. Tryon MD/PhD student Allegheny University of the Health Sciences School of Medicine 3200 Henry Avenue Philadelphia, PA 19129 USA ----------------------------------------- "Quantifying is a committing task." - Cruz-Orive, 1994.
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." - Richard Feynman
} Can anyone explain what the shape factor is (image analyses softwares)
}
} As I understand it, when it is equal to 1 it means the object is a
} disc...what if it is say 2.5 and 0.3??
}
}
} Thanks
}
} F.
Hi,
Shape factors can be really useful for morphometry studies but I've seen some poor formulas used that miss a lot of detail. You have to look at the particular formula to see what the investigator is trying to describe, hopefully, they would give you a few examples of the object of interest and its value so the reader can gauge the meaning and significance of different values.
A good stereology text (unpaid endorsement) that talks a bit about this that I would recommend is:
{fontfamily} {param} Geneva {/param} {bigger} {bigger} Russ, J. C. (1986). {underline} Practical Stereology {/underline} . New York, Plenum Press.
{/bigger} {/bigger} {/fontfamily} I think these journal articles might be helpful too:
{fontfamily} {param} Geneva {/param} {bigger} {bigger} True, L. D. (1996). "Morphometric applications in anatomic pathology [Review]." {underline} Human Pathology {/underline} {bold} 27 {/bold} (5): 450-467.
Royet, J. P. (1991). "Stereology: a method for analyzing images." {underline} Progress in Neurobiology {/underline} {bold} 37 {/bold} (5): 433-74.
Prakash, Y. S., K. G. Smithson, et al. (1993). "Measurements of motorneuron somal volumes using laser confocal microscopy: Comparisons with shape-based stereological estimations." {underline} Neuroimage {/underline} {bold} 1 {/bold} : 95-107.
{/bigger} {/bigger} {/fontfamily} Lastly, a great resource (like this discussion list) for image analysis questions is to go to the NIH-Image website at http://rsb.info.nih.gov/nih-image/, and do a Gopher search of the discussion list archive, which is a terrific resource (plus join the discussion list!). If you didn't know, NIH-Image is free image analysis software, and has a bunch of free macro code that can be easily modified to fit many image analysis needs, and it is still FREE! Yahoo!
good hunting,
{fontfamily} {param} Geneva {/param} Brian C. Tryon
MD/PhD student
Allegheny University of the Health Sciences
School of Medicine
3200 Henry Avenue
Philadelphia, PA 19129
USA
-----------------------------------------
{/fontfamily} "Quantifying is a committing task." - Cruz-Orive, 1994.
"For a successful technology, reality must take precedence over public
relations, for Nature cannot be fooled." - Richard Feynman
At 03:13 PM 9/11/98 +0000, Frank S. wrote: } Can anyone explain what the shape factor is (image analyses softwares) } } As I understand it, when it is equal to 1 it means the object is a } disc...what if it is say 2.5 and 0.3??
It all depends on who wrote the code. Our old LeMont made a point of reporting what parameters were being used to calculate shape. Nowadays it is harder to tell.
Shape is often a ratio of width to length or vice versa. Whether the values are greater than 1 or less than 1 will give you a hint. Then there is the question of how W and L are calculated. They might be minimum and maximum projected measurements. L may be taken as the maximum projected measure and W as the projected measurement at 90 degrees to the L direction. They might be rectangular or elliptical equivalent measures given the features area and perimeter. Or they might be a ratio between perimeter squared and area. And that should, but may not always be, corrected by 4*pi.
Now if you are still using the Visilog package, the answer is Perimeter^2/(4*pi*Area). You should be able to plug in some actual measurements to verify it. A shape of 1 corresponds to a perfect circle - values larger than 1 indicate more elongation and/or convolution. You better not get any (many) values less than 1.
The shape factor of an object is defined by (4 X pi) X area / perimeter squared. In this instance a circle would have a shape factor of 1 and an irregularly shaped object would have a shape factor less than 1. Another way to measure shape is the area perimeter length squared / area. In this latter instance a circle would have a value of 4 x pi (12.57) and irregularly shaped objects would have values greater than this.
John
Can anyone explain what the shape factor is (image analyses softwares)
As I understand it, when it is equal to 1 it means the object is a disc...what if it is say 2.5 and 0.3??
Thanks
F.
John J. Turek, Ph.D. Associate Professor Department of Basic Medical Sciences 1246 Lynn Hall, G193C Purdue University W. Lafayette, IN 47907-1246 Phone: 765-494-5854 Fax: 765-494-0781 Email: jjt-at-vet.purdue.edu
The Chesapeake Society for Microscopy Fall Dinner Meeting
October 13th, 1998 Speaker: Dr. Joseph Gall (after dinner) "Microscopes and Discoveries in Cell Biology: Chicken or Egg?" 6:00 PM Social Hour 7:00 PM Dinner ($20.00, Students $10:00) Make a selection of one at time of reservation Maryland Crab Cakes Assorted Seafood Platter (fried) Filet Mignon with Mushrooms Roast Prime of Beef (thick cut) Pasta vegetarian dish Location: Snyders Willow Grove (410-789-8244) Linthicum, MD Make Reservation and meal selection by Oct 9th To: Andrea Weisberg-(301) 435-1977 aweisberg-at-nih.gov Dinner payable at meeting to 'CSM' Andrea S. Weisberg NIH/NIAID/LVD Bldg.4/Rm.210 4 Center Dr. Bethesda,MD 20892-0445 office (301) 435-1977 Fax (301) 480-1147 e-mail: aweisberg-at-nih.gov
I have an old Balzers CPD, circa 1979. It looks like someone has dropped the cover and chipped the glass window. I am concerned about its safety and need to do something to fix it.
This unit is pretty old and I need some help with repairs or parts. The glass window looks impossible to relace easily, looks like I need a whole new cover. It's a pretty substantial piece of stainless steel, threaded to go on the chamber, with a thick glass window pressed into it.
Does anyone have a lead on getting replacement parts or ideas about a safe repair?
If fixing is not possible, can anyone get me up to date on current CPD's to replace it?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 FAX (831) 429-0146 jmkrupp-at-cats.ucsc.edu
I have a problem of cleaning components of EM like anodes, assemblies and aperatures when they are contaminated. Usual practice is to rub with diamond paste of various grades (depending upon contamination level) and ultrasonicating in solvent like methnol. Finally rinsing three-four times with fresh methonal. One SEM user has suggested me to ultrasonicate the items directly with liquid Ammonia and demonstrated it in his lab. Although it shows a very good cleaning, but I am fraid of whether they really works in EM.
As I have not come across any such reference can anybody experienced in this field, kindly guide me?
Thanks in advance.
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road, Pune - 411 004 India Phone : 91-212-353680/354357 91-212-351542 E-mail: rajdeep-at-aripune.ernet.in
The shape factor is defined as 4 * PI * area / (perimeter^2).
For a circle this computes to 1. Since a circle has the largest area for a given perimeter, any other geometrical figure will have a shape factor smaller than 1. Due to pixelation artifacts, sometimes values of larger than 1 are calculated, but these are artifacts that mostly appear for very small particles.
Hope that helps.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St. # 105N Lakewood, CO 80215 voice: (888) FIND-SIS fax: (303) 234-9271 info-at-soft-imaging.com http://www.soft-imaging.com
} -----Original Message----- } ---------- } From: } "frank.sarrazit-at-AVESTASHEFFIELD.COM"-at-sparc5.microscopy.com[SMTP:"frank.sarra } zit-at-AVESTASHEFFIELD.COM"-at-sparc5.microscopy.com] } Sent: Friday, September 11, 1998 9:13 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Shape factor (image analysis) } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} It all depends on who wrote the code. Our old LeMont made a point of } reporting what parameters were being used to calculate shape. Nowadays it is } harder to tell. snip } Now if you are still using the Visilog package, the answer is } Perimeter^2/(4*pi*Area). You should be able to plug in some actual } measurements to verify it. A shape of 1 corresponds to a perfect circle - } values larger than 1 indicate more elongation and/or convolution. You better } not get any (many) values less than 1.
In addition, we have used a similar 3-dimensional factor derived from Green (1927): sf= (36 x pi x vol-squared)/surface-area-squared for which sphere =1, cube=0.52 and a unit cylinder=0.67
Gib Ahlstrand wrote: } } } Responding to the message of {v04003a09b21dc32600cf-at-[141.233.130.134]} } from "wise-at-vaxa.cis.uwosh.edu"-at-Sparc5.Microscopy.Com: } }
} } The needle valves (vent, fill and drain) on our Ladd critical point drier } } are quite diffucult to turn, even when warm. Does anyone know if they can } } be lubricated and, if so, how? } } I have the same problem with my Ladd CPD. Other than that, I find the Ladd CPD } to be a very good device (IHNCIIL,JASC). I allow users of the CPD to loosen the } valves with pliers, but NOT to tighten them that way, just use finger pressure } to tighten. But I suspect that those with delicate fingers cheat a little when } I'm out of the room, but hopefully, without over-tightening the valves. So far } so good. We've had the unit for over 12 years and never had to replace a valve. } } Is it possible to adjust the turning tension on these needle valves?? } } Gib Ahlstrand
Dear Gib Ahlstrand,
Thank you for your kind comments about the Ladd CPD. You can adjust the tension nut next to the foam behind the needle valves. A word of caution though, you must be careful not to overloosen as it may start to leak. You can also remove the tension nut, take the stem from the valve and apply graphite, white lithium or teflon grease to make the needle valves easier to turn. Too much grease may clog the valve so it should be applied conservativly. If you have any further questions you would like to discuss, please feel free to call Mike Bouchard here at Ladd at 1-802-878-6711. Hope this is of some help,
John Arnott Chairman --
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
I've done cell culture using LRWhite fairly similarly to how you treated your cells. I dehydrated to 100% ethanol, which may have helped with infiltration. I didn't do it, and I don't know if you did, but embedding in a vacuum might help.
When was working on these cells, the only dishes the cells would grow on were slightly soluble in LRWhite. Where the plastic dissolved, it was opaque white, covering the cells. I asked the company if there was a way around that, they said that it shouldn't interfere with anything I was doing. So I sectioned through the dish and the LRWhite, and was able to immunostain. I don't know to what degree, but I guess I embedded my cells in LRWhite and the soluble component of tissue culture plastic. Maybe that stiffened it to prevent the folding problem you're having. Hope this is helpful. Charlie Ginsburg Research Dept. National College of Chiropractic Lombard IL
Linda Fox {lfox1-at-wpo.it.luc.edu} wrote: } } Hello all, } Has anyone experienced puckering of tissues embedded in LRWhite resin? My tissues are cell cultures fixed in 2%PFA, .5%GA, dehydrated to75% ETOH then into graded LRWhite resin. I cut on a diamond knife and embed on 200 mesh grids. Before staining, the puckers and wrinkles are seen as small, and frequen, folds over most cells or along the cell borders. The resin areas are totally free of folding. It's as if the tissue area is picking up H2O during sectioning then has no where to go when it re-dries....very frustrating. After staining it's worse, as the stain seems to stay in the folds and gets really dark. Any thoughts and suggestions are very welcome as always. } Linda Fox } Loyola University } Stritch School of Medicine } lfox1-at-wpo.it.luc.edu }
_________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
Contamination removal has become a very hot topic and has prompted a lot = of research into methods to remove it. We have commercialized a plasma cleaning system based on technology developed by Dr. Nestor Zaluzec (our friendly neighborhood sysop!). We have collaborated on a lot of research=
into plasma cleaning for electron microscopy and have done a fair amount = of work on cleaning microscope parts and accessories. =
While our system was actually developed for cleaning TEM specimens and specimen holders, it can also be used conveniently for cleaning any type = of microscope parts. The only limitation is that it must be able to fit within the 6" ID and 4" tall chamber - not a problem for almost anything you would like to clean. Please contact me off-line or visit our web sit= e for more detailed information.
Elisabeth: Aqueous means water based. So you need to make a 60% and a 5% solution of those chemicals in water. A percent solution of a solid in water solution means somany grams made up to 100 ml. So its 60 grams made up to 100ml when fully dissolved - not many powders will dissolve in that concentration. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au
On Friday, 11 September 1998 11:37, Cox, Elizabeth [SMTP:CoxE-at-prose.dpi.qld.gov.au] wrote: } I have a method for an Oil Red O stain for GMA sections. } It uses 60% } aqueous triethyl phosphate and 5% aqueous ferric ammonium } sulphate, however } I can only locate these chemicals in the powder form. } Does anyone know if } it matters if the powder form is used and what they are } likely to be } dissolved in as it doesn't state this in the methodology?
} Elizabeth Cox } Fisheries Biologist } Queensland Department of Primary Industries } Northern Fisheries Centre, } PO Box 5396, } Cairns Qld Australia 4870 } } ph:+61 7 4035 0100 } Fax: +61 7 4035 1401 } } }
Sally, about 15 years ago somebody published the reason for this Os "pepper". Simply, for this to occur, some chemically unbound Os, GA and phosphate must remain in the tissue. If any one of these is missing you will not get this frustrating artefact.
I suppose this is one major reason for the popularity of cacodylate buffer. It makes the very thorough rinsing process which is required between GA and OS redundant; simply any unbound GA no longer matters.
The situation is aggravated by your fixation schedule. That tissue is overfixed! One hour in 1% GA and one hour in one percent OSO4 in the cold is the general standard for fixing of soft tissues. Intestine should be easy and which animal the tissue is from would not matter for fixation purposes. Fixing at room temperature may be two to four times more severe than fixing "on ice". Your fixation schedule should be close to the point were the osmium completely oxidised membranes and "white" spaces only remain. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Thursday, 10 September 1998 6:13, Sally Burns [SMTP:burnssal-at-pilot.msu.edu] wrote: } Gut tissue fixation } I am looking for a protocol for intestinal tissue. I have } opossum } intestinal tissue fixed by a colleague that has a lot of } OsO4 pepper. I } have also received gut tissue that appeared to have } undergone autolysis. I } am expecting more of these samples and would like to } recommend a change in } protocol. It is probably not possible to use perfusion. } I am looking for intercellular and intracellular bacteria } and /or } protozoal infections in these tissues. } Would the chemistry of the intestine be causing this type } of problem? Or } is it simply a matter of inadequate washing? } Thanks??.. Sally } } The fixation used was: } 2% glut in PBS at pH 7.2 for 3 hours. } 3X washed in PBS } 1X washed in H20 } 1% OsO4 for 4H room T } washed 3x in H20 } dehydrated in a 25% series of acetone } Infiltrated and embedded in a 25% series into a Spurrs/ } Quetol blend. } } } } Sally Burns } Center for Electron Optics } B5 Pesticide Research Center } (517) 355-5004 } } burnssal-at-pilot.msu.edu }
In theory, I expect that when compared with a tungsten filament, at least the solid, single post LaB6 cathode design by Kimball, would make for less movement, especially during the warming-up period. In practise, tungsten filaments are more stable because the emitting area is much larger and so, minor misalignment due to a drifting filament is less consequential. The stability we are talking about is due to slow drift and this is of no consequence to normal, including high resolution imaging in TEM or SEM. Stability over several minutes matters when performing quantitative microanalyses with a probe or EDS in dedicated SEM/TEM. In quantitative analyses a spectrum maybe acquired over two minutes and that is related to standard spectra and all numbers must lead to results that come within 1% of reality. The obvious way to increase stability in LaB6 is to increase the size of the microflat at the tip of the LaB6 cone. The normal size of the cone (Kimball's) is 15 or 20 micrometer. A 40 micrometer microflat is also available (unfortunately at greater cost) and this makes a LaB6 quite suitable for quantitative microanalyses. Brightness is reduced but for microanalyses that is no consideration. Reducing downtime by increasing "filament life" to over 5000 hours at low and medium emission is the main benefit of a Lab6 in a microprobe. For non-analyses work, a standard flat Lab6 cathode is quite stable for SEM and TEM requirements; extreme drift would matter but stability worries in these applications are more likely to relate to HV instabilities or a pulsing beam due to column/aperture contaminations. The question for Mark Darus is the EM's vacuum system: Is it good enough for LaB6 operation. Disclaimer: ProSciTech supplies Kimball cathodes and filaments. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Wednesday, 9 September 1998 0:38, Mark Darus [SMTP:DARUSM-at-cle.lg.bfg.com] wrote: } I'm new at operating an SEM that has a tungsten filament. } For the past 5 } years I had an instrument that used a LaB6, but I've } changed companies. } I have 2 questions. } 1. Both the people here and the instrument's service man } tell me } that the tungsten filament is more stable than the LaB6. } I ask them to } explain further and they really don't get into it. What } is the stable thing } about the tungsten filament? My LaB6 seemed fine to me, } perfect in fact, } so what is the unstableness of it? } 2. I'm also told here that, with the tungsten filament, } as the beam sits in } one area on the sample, Carbon will develop in that area. } Is this true? If } so how long does it take for the carbon to contaminate the } area, and also } does this take any confidence in a carbon analysis and } throw it out the } window? }
Just a reminder to all that, one of the good places to look for things like this is the SUSTAINING MEMBERS WWW page of MSA
http://www.msa.microscopy.com/SM/SustMembers.html
from that page the Current GW address is:
GW Electronics, Inc. Attn: Mr. Larry H. Glassman 6981 Peachtree Industrial Blvd. Norcross, GA 30092 Phone(days): (404) 449-0707 Fax Number: (404) 449-0284 E-Mail:
} } I need to get in touch with a company that used to be called GW } Electronics and they used to be in Norcross, Ga. My "new address and } phone number" card is dated Sept. 1, 1983. Are they still around? } } Bill } -- } ============================================================= } Bill Chissoe III } Electron Microscopist,University of Oklahoma } E-mail: wchiss-at-ou.edu Ph. (405)325-4391 } =============================================================
At 03:25 PM 9/3/98 +0900, =C1=A4=C1=F6=C8=C6 wrote:=20
} } } }
{excerpt} {fontfamily} {param} =B1=BC=B8=B2 {/param} {smaller} I am a freshman stu= dying dentistry in Seoul, Korea. Yesterday my professor gave us a question which we were supposed to answer via e-mail as quick as we could. The problem was "Why is there the 'phase contrast' in PCM?" I looked up all the references available to me, and searched all over the internet for 5 hours, and still don't have the answer. If can anybody help me, please e-mail me at { {mailto:gehoon-at-plaza1.snu.ac.kr} gehoon-at-plaza1.snu.ac.kr
{/smaller} {/fontfamily}
{/excerpt} { { { { { { { {
Hi,
The answer is yes, there very definitely is phase contrast in PCM. In regular brightfield microscopy, the image is formed by interference between the undiffracted background light and the light diffracted by the features in the specimen. This interference occurs at the Primary Image Plane, which can be easily viewed by removing the eyepiece and stretching a piece of lens paper over the resulting opening. The problem in viewing many unstained biological samples is that the phase relationship between the undiffracted background light and the diffracted specimen light is on the order of a quarter of a wavelength or less. The result is incomplete interference and very low contrast.
For Phase Contrast Microscopy, we carefully engineer the microscope to take advantage of this quarter wavelength phase shift. The whole concept is based on the the principle that, when waves are HALF a wavelength out of step, they will undergo destructive interference. Our challenge: to increase the phase shift between the background light and the specimen light to meet this requirement. The implementation is elegantly simple (so elegant, it earned Frits Zernike the Nobel prize!):
a. First, we need to control the exact location of the background light.=20 To accomplish this feat, we limit the aperture in the front focal plane of the condenser to just a ring.
b. Next, we need to generate the extra quarter wave difference. We accomplish this feat by inserting a special phase plate in the back focal plane of the objective. You can view this plane by rotating a phase objective in place then removing the eyepiece and looking far down the tube, into the back of the objective. There you will find a dark or "smokey" ring. =20
This back focal plane is "conjugate" (optically related) to the front focal plane of the condenser. If you rotate the phase annulus in the condenser into position, you will see the bright ring underlaying the smokey one. When the phase system is correctly aligned, you will notice that the bright ring from the condenser is imaged precisely in the darker ring mounted in the objective.=20
The phase plate which is mounted in the back focal plane of the objective has two characteristics:
One is a {underline} channel {/underline} , usually of less thickness than the general area of the plate. In most conventional phase systems, this channel is cut into the ring so that the background light has less glass to go through. The amount of the cut allows the background light to gain the extra quarter wave jump on the diffracted light from the sample.
The second characteristic of the phase plate is a {underline} neutral density filter {/underline} (the reason that this ring looks dark). For optimum interference, the two interfering waves need to be approximately the same amplitude. However, the diffracted light is usually only about 15% as bright as the background light. To correct this mismatch, we coat the cut through which the background light passes with a neutral density material.
The final outcome: when the diffracted light from the specimen meets the undiffracted light from the background in the Primary Image Plane, they are out of step by a total of one-half wave and are about the same intensity. They destructively interfere, generating darker features against a lighter (soft gray) background. Voila! Phase contrast.
c. One more point: To make the system work really well, you need to take into account several variables.
One is the sample: it really needs to be the type which closely approximates the initial quarter wavelength phase shift. Since this shift depends on both the real, geometric thickness of the specimen as well as the difference=20
in refractive index between the mounting medium and the sample, you can fine tune the system by changing the mounting medium (try glycerin, white corn syrup, or even immersion oil, if you sample will tolerate=20 it)
The second is the wavelength of light. Notice that we did not mention a quarter of a wave shift for any particular wavelength. Most modern phase contrast microscopes are engineered for 546 nm, so you need to use a good 546 nm (rich green) filter.
Finally, alignment is critical. Make sure that you have the correct phase ring or annulus in the condenser which matches the phase plate in the objective.
There are several really good references on Phase Contrast:
1. Ross, K. F. A. Phase Contrase and Interference Microscopy for Cell Biologists. Edward Arnold, Ltd, London. 1967
2. Pluta, M. Advanced Light Microscopy, Vol. 2.,Elsevier, NY. 1988
For a thorough but practical discussion, we also recommend our book, Optimizing Light Microscopy for Biological and Clinica Laboratories (1997). Details are available on our web site: { {http://www.MME-Microscopy.com/education}
Augusto, please check Micron 27(2) 1997, 129-139 and J. of Microscopy 188, 1997, 285-289.
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Fri, 11 Sep 1998 Augusto_A_Morrone-at-notes.seagate.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Subject: TEM imaging, CCD Vs film } } There have been several threads in the past on the advantages and } disadvantages of film and CCD cameras to acquire images in the TEM. I } enjoy having both capabilities as complementary in the operation of the } TEM, and became dependent on both. For convenience, I still take images } on film and then digitize the negatives; sometimes I also make contact } prints or 5x to 9x enlargements in the darkroom. However, I have no hard } data (numbers) to back up my impression that the resolution and contrast } range of film is better. Can anyone send me a note on this issue, a } reference, or forward me an earlier posting (couldn't find it in the } archives for the last several months)? } } Thank you. } } Augusto A. Morrone } Seagate Technology } 7801 Computer Ave South } Bloomington, MN 55435-5489 } Phone: (612) 844-5838 } Fax: (612) 844-8247 } Augusto_ A_Morrone-at-notes.seagate.com } } }
Augusto, please check Micron 27(2) 1997, 129-139 and J. of Microscopy 188, 1997, 285-289.
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Fri, 11 Sep 1998 Augusto_A_Morrone-at-notes.seagate.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Subject: TEM imaging, CCD Vs film } } There have been several threads in the past on the advantages and } disadvantages of film and CCD cameras to acquire images in the TEM. I } enjoy having both capabilities as complementary in the operation of the } TEM, and became dependent on both. For convenience, I still take images } on film and then digitize the negatives; sometimes I also make contact } prints or 5x to 9x enlargements in the darkroom. However, I have no hard } data (numbers) to back up my impression that the resolution and contrast } range of film is better. Can anyone send me a note on this issue, a } reference, or forward me an earlier posting (couldn't find it in the } archives for the last several months)? } } Thank you. } } Augusto A. Morrone } Seagate Technology } 7801 Computer Ave South } Bloomington, MN 55435-5489 } Phone: (612) 844-5838 } Fax: (612) 844-8247 } Augusto_ A_Morrone-at-notes.seagate.com } } }
Augusto, please check Micron 27(2) 1997, 129-139 and J. of Microscopy 188, 1997, 285-289.
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Fri, 11 Sep 1998 Augusto_A_Morrone-at-notes.seagate.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Subject: TEM imaging, CCD Vs film } } There have been several threads in the past on the advantages and } disadvantages of film and CCD cameras to acquire images in the TEM. I } enjoy having both capabilities as complementary in the operation of the } TEM, and became dependent on both. For convenience, I still take images } on film and then digitize the negatives; sometimes I also make contact } prints or 5x to 9x enlargements in the darkroom. However, I have no hard } data (numbers) to back up my impression that the resolution and contrast } range of film is better. Can anyone send me a note on this issue, a } reference, or forward me an earlier posting (couldn't find it in the } archives for the last several months)? } } Thank you. } } Augusto A. Morrone } Seagate Technology } 7801 Computer Ave South } Bloomington, MN 55435-5489 } Phone: (612) 844-5838 } Fax: (612) 844-8247 } Augusto_ A_Morrone-at-notes.seagate.com } } }
Bill Chissoe III wrote: ================================================== I need to get in touch with a company that used to be called GW Electronics and they used to be in Norcross, Ga. My "new address and phone number" card is dated Sept. 1, 1983. Are they still around? ================================================== G-W Electronics is very much "still around". Their website is at URL http://www.gwelectronics.com/
and their address/contact information is the following:
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Does anyone know of a source for an old Spencer-style external lamp with collector lens, diaphragm, filter holder, and bulb centering adjustment? Thanks.
-----Original Message----- } From: System Administrator To: Howe L. C. Josephine Sent: 9/13/98 11:24:59 AM
} ---------- } From: Howe L. C. Josephine } Sent: Sunday, September 13, 1998 11:24 AM } To: 'microscopy-at-msa. microscopy.com' } Subject: radiation from uranyl acetate } } A postgraduate had prepared 3 tubes of 10g uranyl acetate in 15 mls water. } Later she decided to dispose it away. She brought it to the radiactive } waste disposal to discard it. The safety officer did a check to see } whether it was safe to dispose it there. The radiation emitted was very } high, about 500 counts per unit. She was told it was not safe for her to } handle it without protection. She was worried and went for a blood test. } It showed the cell count of lymphocyte was lower than normal. 6 weeks } later she went for another count. This time the cell count was much } higher. She is now very worried and would like to how harmful is the } radiation from uranyl acetate. Can anybody help to ease her anxiety? } Till today she has not forgiven people who has been handling uranyl } acetate for not informing her of the risk. } } Josephine Howe } NUS }
} Does anayone on this network have experience with the new field of Raman } Imaging ? We have a raman instrument with a tunable filter and imaging. I } have a couple of questions. I am a TEM person and new at this so excuse } me if these questions are too elementary. } } The manufacturer claims that on aquiring the image the spatial resolution } of the 'spectra' is as small as a pixel. But since unlike IR, Raman is a } scattering process wouldn't all regions limited by the beam (probe) , } about 150 microns, be affected by each other and thus negating the claim } of 1 pixel resolution? Infact I did observe this experimentaly while } imaging composite polymeric samples. Spectra from all regions illuminated } by the beam are almost identical although the image } does exhibit contrast due to different phases.
This question can be best answered by the manufacturer of your instrument. However, I guess that your instrument has been designed to get the best possible resolution. In that case, as in other optical microscopy, the resolution is limited by the diffraction of light. In other words, due to the wave nature of light you can not focus a light beam below a certain size. In the visible range that means that your spatial resolution is around one micrometer on the sample.
Probably the manufacturer has arranged things in such a way that one micron on the sample is imaged at least as one pixel on the CCD detector.
I hope this helps.
--------------------------------------------------------------------------- Fernando Agullo'-Rueda Raman Microscopy Laboratory Instituto de Ciencia de Materiales de Madrid (CSIC) Cantoblanco, E-28049 Madrid Espan~a (Spain)
We normally use 'Quadralene' which is a detergent containing ammonia (followed by rinsing), although I have used ammonia when cleaning gun parts which aren't easy to remove from the gun. The only thing to remember is that you must never use ammonia on copper or brass parts (if you have any in your gun). Also I am surprised that you use methanol for cleaning - do you not think that ethanol or acetone are safer?
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: Rajdeep Dongre To: microscopy
Dear Friends,
I have a problem of cleaning components of EM like anodes, assemblies and aperatures when they are contaminated. Usual practice is to rub with diamond paste of various grades (depending upon contamination level) and ultrasonicating in solvent like methnol. Finally rinsing three-four times with fresh methonal. One SEM user has suggested me to ultrasonicate the items directly with liquid Ammonia and demonstrated it in his lab. Although it shows a very good cleaning, but I am fraid of whether they really works in EM.
As I have not come across any such reference can anybody experienced in this field, kindly guide me?
Thanks in advance.
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road, Pune - 411 004 India Phone : 91-212-353680/354357 91-212-351542 E-mail: rajdeep-at-aripune.ernet.in
id AA01120; Mon, 14 Sep 1998 10:12:55 -0400 Received: from FETC-Message_Server by FETC.DOE.GOV with Novell_GroupWise; Mon, 14 Sep 1998 10:16:27 -0400 Message-Id: {s5fcecfb.080-at-FETC.DOE.GOV} X-Mailer: Novell GroupWise 4.1
I have a way of doing this that I use regularly with good results in my study coal ash deposits---similar to rocks and fracture surfaces in ceramics. I use reflected-light illumination (bright- and dark- field) . The method can be easily implemented with typical image processing software---I have Optimas macros available. For more information: ftp://titan.petc.doe.gov/pub/ramer (Under /docs is a Word document of my Microscopy Today article [Feb/March 98], a PowerPoint file illustrating results, and an AVI file of an animated fly-by of a dot on the back of a penny. The macros are under /Optimas.)
Briefly, the method is: 1. Get a stack of images---The raised dot on a coin makes a good object: I use 5 um steps with a 20X objective and 2 um for 50X, which can be done easily by hand. 2. Save the focused part of each image---Apply the LAPLACIAN operator to find texture (scratches). Convert the result to a binary image (choosing the threshold will require some trial and error, but the threshold will remain the same for all the images in the stack). The binary image will have "snowy" patches corresponding to the regions with texture. Merge the "snow flakes" into solid patches by DILATING several times. AND the result with the original image and you should have the focused part. 3. Combine the focused parts of all the images---Use the MAX operator instead of ADD (or OR) because the focused parts might overlap some.
Everett Ramer Federal Energy Technology Center Pittsburgh, PA (412)892-4920
The ChemIcon instrument uses the LCTF as a tunable bandpass filter. The idea behind the instrument is to globally illuminate the sample with a large laser spot and then collect all of the Raman scatter from the sample and filter it through the LCTF.
You then can collect spectra by taking a series of images with the CCD, each at a different wavelength. If you select the same pixel in each of the wavelength dependent images and plot it's intensity as a function of the image wavelength, you can reproduce a spectra. Therefore, if you have a 100 pixel by 100 pixel image, you can have 10000 spectra in one spectral image dataset. The spectral resolution is based on the bandpass of the filter, which is about 7 cm-1. The spatial resolution, as Fernando said, is diffraction limited.
Although I no longer work for the compant, I helped design the ChemIcon instrument. So, if you have any questions about it's use or the science behind it's design, please feel free to contact me.
Tim Prusnick, PRUSNICK_TIM-at-worldnet.att.net Raman Application & Support Engineer Renishaw Inc. 623 Cooper Court Schaumburg, IL 60173 PHONE: 847-843-3666 FAX: 847-843-1744
-----Original Message----- } From: Fernando Agullo-Rueda [mailto:FAR-at-icmm.csic.es] Sent: Monday, September 14, 1998 2:35 AM To: microscopy-at-Sparc5.Microscopy.Com
Kalpana S Katti wrote:
} Does anayone on this network have experience with the new field of Raman } Imaging ? We have a raman instrument with a tunable filter and imaging. I } have a couple of questions. I am a TEM person and new at this so excuse } me if these questions are too elementary. } } The manufacturer claims that on aquiring the image the spatial resolution } of the 'spectra' is as small as a pixel. But since unlike IR, Raman is a } scattering process wouldn't all regions limited by the beam (probe) , } about 150 microns, be affected by each other and thus negating the claim } of 1 pixel resolution? Infact I did observe this experimentaly while } imaging composite polymeric samples. Spectra from all regions illuminated } by the beam are almost identical although the image } does exhibit contrast due to different phases.
This question can be best answered by the manufacturer of your instrument. However, I guess that your instrument has been designed to get the best possible resolution. In that case, as in other optical microscopy, the resolution is limited by the diffraction of light. In other words, due to the wave nature of light you can not focus a light beam below a certain size. In the visible range that means that your spatial resolution is around one micrometer on the sample.
Probably the manufacturer has arranged things in such a way that one micron on the sample is imaged at least as one pixel on the CCD detector.
I hope this helps.
--------------------------------------------------------------------------- Fernando Agullo'-Rueda Raman Microscopy Laboratory Instituto de Ciencia de Materiales de Madrid (CSIC) Cantoblanco, E-28049 Madrid Espan~a (Spain)
Dear Rajdeep, } } I have a problem of cleaning components of EM like anodes, assemblies } and aperatures when they are contaminated. Usual practice is to rub with } diamond paste of various grades (depending upon contamination level) and } ultrasonicating in solvent like methnol. Finally rinsing three-four times } with fresh methonal. One SEM user has suggested me to ultrasonicate the } items directly with liquid Ammonia and demonstrated it in his lab. } Although it shows a very good cleaning, but I am fraid of whether they } really works in EM. } We remove some of the harder-to-clean contamination with a polishing compound (similar to your diamond paste method), then we soak the parts-- mostly aluminum liners, stainless steel Wehnelt cylinders, and a few of other materials--in Alconox, a mild detergent, rinse with distilled water, rinse with ultra-pure water, rinse with ethanol (better for fingerprints & less toxic than methanol), and rinse with acetone. We do not sonicate; it is not a safe procedure with organic solvents, but most people get away with it. The microscopy list archives have considerable information about this. Good luck. Yours, Bill Tivol
Ruthenium tetroxide is a choice for polystyrene staining and since it is a strong oxidizing agent you will need to do a 1, 2, 3....min stainig time to determine the best time period for your sample. Be aware that the staining needs to be done under the hood-follow safety procedures- If you have any specific questions email me.
Ani
Ani M Issaian California Institute of Technology Pasadena, CA. 91125 MC 210-41
I am interested in high resolution coaters to use with field emission SEM. Does anyone working with FE-SEM's have some opinions for me on what is best specifically for polymer applications. Steve
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
Anyone doing yeast embedding for TEM immunocytochemistry on a routine basis, please contact me directly to discuss a possible collaborative work. Thanks in advance.
Michel **************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
As most of the people on the web know Protrain have a history in EM servi= ce and operator training and that we regulary run courses on the maintenance=
of electron microscopes. =
A little plug for the University of Sydney, Australian Centre for Microscopy and Analysis, who will be hosting our "Monitoring & Maintainin= g the Electron Microscope" course from 26th to 29th October 1998. Contact = - emma-at-emu.usyd.edu.au - for details.
That said attached is a cleaning programme for electron microscopes that = I hope will help you out.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Maintaining a Scanning Electron Microscope=0D =0D TUNGSTEN Gun Systems=0D =0D The cathode assembly should be cleaned every filament change, the anode e= very other change and the electron gun at least once a year.=0D =0D Materials - Almost any metal polish may be used to clean electron gun com= ponents however it must not be LONG LIFE. Long life additives coat the c= leaned item with a polymer that causes chaos in the electron gun. Look o= ut for any indication on the bottle or tube that the manufacturer is clai= ming that you will not need to clean the metalwork so often after using t= heir product!=0D =0D Method - Almost more important than the cleaning efficiency is our abilit= y to completely remove the polishing media. So many service call outs ar= e due to problems caused through inefficient removal of the media. For t= his reason it makes sense to use a metal polish that is easily removed by= a solvent for tungsten. In this way we not only remove the metal polish= but also clean the areas that are difficult to approach with the polish,= nooks and crannies! Also very important is the need to clean without da= maging the component, scratching it or placing cotton hairs within the "t= raps" that the manufacturers seem to put in our way. The best cleaning t= echnique is a wet clean, which is to use solutions and an ultrasonic clea= ner. In this way the damage that mechanical forces apply to the componen= ts are minimised. Sure the cathode aperture may need a little more encou= ragement to give up its deposit but only do this if the wet cleaning proc= edure falls short. We like "Silvo" or "Bluebell" or "Brasso", liquid met= al polishes that will mix with a dilute ammonia solution to form a cleani= ng media, but a solution that may be removed with further washes in dilut= e ammonia. The mix - 10% metal polish in 90% ammonia solution - where th= e solution is 10% ammonium hydroxide in water. Place the components, one= at a time, in the solution with their least important face down wards. = Never put gun components together in the solution, as they will damage ea= ch other. Do not put an aluminium cathode in ammonia as it will go black= , oxide! After 20 minutes in an ultrasonic the component should be clean= , wash off in running water and run for another 5 minutes in straight 10%= ammonium hydroxide in water. Swill off with running water and then wash= in alcohol and dry. NEVER throw away your solutions until you have reas= sembled the cathode, as it is quite possible for the small screws to have= fallen out and to reside in the debris at the base of one of the cleanin= g containers. If you do have a deposit remaining in the aperture area of= the cathode a little mechanical effort with the cleaning media may be re= quired.=0D =0D Once a component is clean check it with a hand lens or binocular microsco= pe before returning it to the microscope OR wrap it in kitchen (aluminium= ) foil until required. When working with clean components work on a shee= t of this foil as it is very clean and it makes an ideal working surface.= =0D =0D When rebuilding the components and placing them in the microscope try to = place the higher components first so that you are less likely to drop deb= ris on the cleaned components below. Alternatively always cover the colu= mn with aluminium foil when it is opened for removing components or close= the gun chamber down whilst cleaning is being carried out.=0D =0D The gun chamber IS important and this should be cleaned through disassemb= ly once a year, particularly with a TEM. Dirty guns hold gas and induce = micro discharge, which spoils images. Clean the gun chamber with metal p= olish, remove the metal polish with dilute ammonia and buff up the walls = with a clean chamois or dear skin leather. To retain the cleanliness of = the chamber, each time you change a filament buff up the walls with the l= eather. If the chamber smells, oily-ozone smell, but is not visibly stai= ned, this is the result of discharge and all traces of the smell should b= e removed with dilute ammonia.=0D =0D Look after your gun, it is probably the dirtiest area of the microscope, = other than the specimen area in a SEM or the camera chamber in a TEM, its= state will determine the ultimate performance of the instrument and your= filament life.=0D =0D LANTHANOM HEXABORIDE=0D Technique developed by Biology E.M. Unit Canberra=0D =0D Clean the cathode with 25% hydrochloric acid in water by immersing for 60= seconds and then cleaning with a weak alkaline (ammonia or sodium hydrox= ide). Wash with water and then alcohol before drying.=0D =0D LaB6 sources should last a long time (1000 hours plus) but they do need a= n intermediate cleaning session about every 250 to 350 hours. Some peopl= e amaze us by getting away with 1100 hours without cleaning but this is t= he exception not the rule.=0D =0D THE ELECTRON COLUMN=0D =0D The column requires cleaning when you find the stigmator controls reach t= he end of their range. In the first case change the variable aperture (f= inal aperture) to see if that enables you to carry on working within the = stigmator range. If so then you know that the aperture you first used is= too dirty for the kV you intended to use. If changing the aperture does= not change the level of astigmatism the problem is in the rest of the co= lumn; you have no alternative but to clean it.=0D =0D The column liner may be removed from underneath the anode or on some inst= ruments from the specimen chamber downwards. Some of the Hitachi range h= ave column components that are removed from the gun chamber end as well a= s the specimen area and complex manipulations may be required (watch your= service engineer).=0D =0D Clean the column liner with an Ultrasonic cleaner if possible, as a "wet"= clean is better at getting down inside the tubes. If the cleaner is amm= onia based it will attack copper-based materials so do not leave them in = the cleaner for more than a few minutes. Straight 5% ammonia solution in= water is fine but if you need a little abrasion to help the process the = commercially available "Quadralene" is ideal.=0D =0D The ultrasonic solution described in the gun cleaning process is fine pro= vided you do not leave the components in the ammonia solutions for more t= han a few minutes.=0D =0D SHINY APERTURES=0D =0D The silver coloured apertures are made of molybdenum or platinum and are = most efficiently cleaned using heat where temperatures in the orange-red = range are required.=0D =0D The ideal cleaning procedure uses a high vacuum coating unit where the ap= ertures are placed upon a platinum (for platinum) or molybdenum (for moly= bdenum) boat. Current is passed through the boat under high vacuum holdi= ng the apertures at orange-red until they display a constant colour all a= cross the aperture. Do not look at the boat without dark glasses and do = not let the temperature drift into the white range or you may melt the ap= ertures. After cleaning check with a lens that the apertures are still p= erfectly round. Throw miss shaped apertures away as they will give astig= matism problems if used.=0D =0D If you do not have a high vacuum coater you may clean platinum apertures = by holding them in a Bunsen flame using a platinum boat or platinum tippe= d tweezers. Again go to orange-red heat until the aperture glows one col= our all the way across. If you try to heat up molybdenum apertures in th= is way they will oxidise and go black. You have no alternative with thes= e apertures but to replace them.=0D =0D After cleaning any component check it thoroughly with a lens before placi= ng it back in the microscope. Always have a stock of the metal apertures= ready for replacement.=0D =0D VACUUM SEALS=0D =0D Each time you find a vacuum seal this requires some action. Remove the s= eal but ONLY USE A WOODEN STICK FOR REMOVAL. Gently pull the "O" ring th= rough the groove between the base of your thumb and first finger. This s= hould remove any debris and the "finger grease" should be sufficient to l= ubricate the "O" ring. DO NOT GREASE AN "O" RING UNLESS IT IS A MOVING S= EAL. Do not clean the "O" rings in a solvent as they will dry and start = to crack after repeated cleaning. If an "O" ring is really dirty wash it= in hot soapy water, running it between your fingers to massage in the cl= eaning media.=0D =0D Clean the "O" ring seat with a gentle solvent like alcohol before replaci= ng the seal.=0D Protrain Maintaining a Scanning Electron Microscope 3=0D =0D
} A postgraduate had prepared 3 tubes of 10g uranyl acetate in 15 mls water.
This is more UA than we usually use--our max is 1 or 2 g for 100 ml of 1% or 2% stock solutions.
} Later she decided to dispose it away. She brought it to the radiactive waste } disposal to discard it. The safety officer did a check to see whether it was } safe to dispose it there. The radiation emitted was very high, about 500 } counts per unit.
It is important to know what are "counts per unit". If you mean counts per second, that is a moderate amount of counts. It is also im- portant to know how the measurement was obtained. If a Geiger counter was used and the counts taken outside a closed jar, they likely arise from gamma rays, which will penetrate the skin and cause damage. If the counts were taken with a liquid scintillator, they could also arise from alpha particles, which are the decay product of the U, or beta particles, which are from decay of a daughter nuclide. Alphas and low-energy betas will not penetrate the dead layer of the skin. They are not harmful unless the material is inhaled, ingested or absorbed through the skin. The last process is not a problem with UA.
} She was told it was not safe for her to handle it without } protection. She was worried and went for a blood test. It showed the cell } count of lymphocyte was lower than normal. 6 weeks later she went for } another count. This time the cell count was much higher.
These counts are not specific for radiation. It is very unlikely that they are related to radiation, and much more likely that they are related to the stress from worrying about radiation. The amounts of radiation which lead to rapid changes in blood counts are massive--drin- king 30 g of UA might lead to that much radiation, and there would be chemical effects from that much UA which could cause blood cell changes.
} She is now very } worried and would like to how harmful is the radiation from uranyl acetate. } Can anybody help to ease her anxiety?
Unless the UA enters the body--as opposed to being on the skin or outside--only the gamma rays from decay of daughter products will be at all harmful. One should always wash one's hands after using UA to remove any droplets which may have gotten on the skin--especially before eating or smoking, which could lead to ingestion of the UA. The key measure for biological effects is the rem (Radiation Equivalent in Man) which is the product of the quality factor--1 for gammas & high-energy betas, about 2 for low-energy betas, and 10 to 20 for alphas--times the dose in rad (Radiation Absorbed Dose). The dose is measured in ergs per gram of tissue with 100 erg/g = 1 rad. This unit is roughly related to a mea- sure of ionization produced by a radiation field (which is measured in Roentgen units). To give an idea of possibilities for harm, the normal background radiation is about 50 mr (millirad) per year, the general population is given a limit of 500 mr per year before exposure is considered to be a problem, and radiation workers are allowed 5 rad per year. Your student should ask the safety officer how many rads were in the 30 g of UA. She should also ask how many mr/hr were measured at the outside of the jars. If this is a high number, she should calculate what she was exposed to. In particular, she should assume all the radiation would be absorbed in her hands. This calculation should reassure her.
} Till today she has not forgiven people who has been handling uranyl acetate } for not informing of the risk. } She should have been informed about the risks--everyone here who uses radioisotopes or radiation-producing equipment has to take a safety course and frequent refresher courses. UA is usually considered to be a negligable risk when used in the amounts usual for electron microscopy. There are far more serious hazards, such as OsO4 and glutaraldehyde, associated with EM. I hope this will put her mind at ease. Yours, Bill Tivol
I'm curious about using a LCTF to filter the laser. All the LCTFs I have seen have a very poor transmissivity. Doesn't this make a large impact on the spectra mwhich is collected by the CCD?
Timothy M. Prusnick wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } The ChemIcon instrument uses the LCTF as a tunable bandpass filter. The } idea behind the instrument is to globally illuminate the sample with a large } laser spot and then collect all of the Raman scatter from the sample and } filter it through the LCTF. } } You then can collect spectra by taking a series of images with the CCD, each } at a different wavelength. If you select the same pixel in each of the } wavelength dependent images and plot it's intensity as a function of the } image wavelength, you can reproduce a spectra. Therefore, if you have a 100 } pixel by 100 pixel image, you can have 10000 spectra in one spectral image } dataset. The spectral resolution is based on the bandpass of the filter, } which is about 7 cm-1. The spatial resolution, as Fernando said, is } diffraction limited. } } Although I no longer work for the compant, I helped design the ChemIcon } instrument. So, if you have any questions about it's use or the science } behind it's design, please feel free to contact me. } } Tim Prusnick, PRUSNICK_TIM-at-worldnet.att.net } Raman Application & Support Engineer } Renishaw Inc. } 623 Cooper Court } Schaumburg, IL 60173 } PHONE: 847-843-3666 } FAX: 847-843-1744 } } -----Original Message----- } } From: Fernando Agullo-Rueda [mailto:FAR-at-icmm.csic.es] } Sent: Monday, September 14, 1998 2:35 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Raman imaging } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Kalpana S Katti wrote: } } } Does anayone on this network have experience with the new field of Raman } } Imaging ? We have a raman instrument with a tunable filter and imaging. I } } have a couple of questions. I am a TEM person and new at this so excuse } } me if these questions are too elementary. } } } } The manufacturer claims that on aquiring the image the spatial resolution } } of the 'spectra' is as small as a pixel. But since unlike IR, Raman is a } } scattering process wouldn't all regions limited by the beam (probe) , } } about 150 microns, be affected by each other and thus negating the claim } } of 1 pixel resolution? Infact I did observe this experimentaly while } } imaging composite polymeric samples. Spectra from all regions illuminated } } by the beam are almost identical although the image } } does exhibit contrast due to different phases. } } This question can be best answered by the manufacturer of } your instrument. However, I guess that your instrument has been } designed to get the best possible resolution. In that case, } as in other optical microscopy, the resolution is limited } by the diffraction of light. In other words, due to the wave nature } of light you can not focus a light beam below a certain size. } In the visible range that means that your spatial resolution } is around one micrometer on the sample. } } Probably the manufacturer has arranged things in such a way that } one micron on the sample is imaged at least as one pixel on the } CCD detector. } } I hope this helps. } } --------------------------------------------------------------------------- } Fernando Agullo'-Rueda } Raman Microscopy Laboratory } Instituto de Ciencia de Materiales de Madrid (CSIC) } Cantoblanco, E-28049 Madrid } Espan~a (Spain) } } Tel: +34-91-334-9015 E-mail: {FAR-at-icmm.csic.es} } Fax: +34-91-372-0623 {http://www.icmm.csic.es/} } {http://www.icmm.csic.es/Fagullo/Fagullo.htm} } ---------------------------------------------------------------------------
-- ****************************** Jim Haley Applications Engineer I-CUBE 2411 Crofton Lane, Suite 14A Crofton, MD 21114 voice: (301) 858-0505 fax: (301) 858-0615 web site: http://www.i-cubeinc.com e-mail: haley-at-i-cubeinc.com ******************************
We have an environmental SEM (ESEM model 2020) with the standard stage. It would be of great help to several of our programs to have a stage with a wider range of motions. ESEM did make and deliver stages with longer traverses. The stage we have moves only about plus or minus one inch (twenty five mm).
If you have an ESEM 2020 with the large traverse stage and are willing to part with the stage, please contact me. We would be willing to buy the stage (if a price can be agreed). We could consider exchanging our stage for yours, if the smaller motion will meet your needs.
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I just began using a Zeiss 10C TEM which has been used by many people before, who unfortunately are no longer here. We have a minor (?) problem with the filament, what appears to be a loose connection of some sort. When the filament is switched on the voltmeter jumps as usual to around 1.5V, but quickly drops to around 0.2-0.3V. The vacuum is fine, and so I'm led to believe it is a loose connection, or the filament has been comprimised otherwise. The service representative has not responded for a few weeks. IF anyone could offer some advice as to what exactly should be looked for in the assembly, to minimize troubleshooting time, it would be very helpful. The professor I'm working for is also new to the university and would rather to have a list of "quick options" than to take the casing apart and so forth.
TJ LaFave University of North Carolina--Charlotte Department of Physics [Department of Electrical Engineering] Charlotte, NC 28223
The Cornell Nanofabrication Facility has an open staff position for electron beam lithography and related microfabrication technologies. The position is available immediately. BS., MS, or PhD.
If you know of anyone interested, please have them contact me.
Thanks
Lynn Rathbun
***************************************************** Dr. Lynn Rathbun, User Program Manager Voice (607)-255-2329 ext 110 Cornell Nanofabrication Facility FAX(607)-255-8601 Knight Laboratory-Cornell University email Rathbun-at-cnf.cornell.edu Ithaca, New York 14853 http://www.cnf.cornell.edu/ Webmaster -at- Christian World Adoption http://www.cwa.org/ Webmaster -at- Joint Council on International Children's Services www.jcics.org (any opinions or representations of fact re: adoption are my own however) *****************************************************
Does anyone know the email address or phone number for
Vishwas Bhide ?
At one time he worked at Intel, but apparently we have lost track of him.
You can email me directly if you wish.
Thanks in advance
Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
we use chromium exclusively for coating our polymer membranes etc. for FESEM work (S900, S4500).
The thing is you MUST pump away every gas other than the sputtering gas, as Cr, unlike Au, will form nitrides & oxides which are not as dense as the metal as a coating on your specimen.
Our coater is the Xenosput, obtained through Edwards. It uses xenon as the sputtering gas. One (small) cylinder lasts ~ 10 years. The best part of its operation is that the final purging of gases in the chamber is achieved by actually sputtering titanium in the chamber itself so you only have xenon left for the final coating stage. Ours has a rotating stage and gives a very even coat over at least 75mm radius.
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
I am new to EMs in general and have begun work on a Zeiss 10C TEM. The filament worked properly for about a week. Now, however, when the filament is activated the voltmeter immediately jumps to 1.5V (as normal), but instead of remaining there it quickly loses potential to about 0.2-0.3V. The vacuum is fine. I suspect a loose connection, but opt for an experienced person to perhaps suggest where I might begin. I am still asking around for the schematics. It shouldn't be a very difficult job (?).
Unfortunately, no one here currently knows the electronic configuration for the filament, and we prefer to know this before going through a series of unguided troubleshooting. Also, the service rep hasn't responded for some two weeks yet. Furthermore, the maintenance section of the manual is missing.
If anyone has had experience and is willing to take the time to offer some useful suggestions it would be greatly appreciated.
TJ LaFave University of North Carolina at Charlotte Department of Physics Charlotte, NC 28223 (704)547-3392
The LCTF used in the ChemIcon instrument has a transmissivity max of only 20-30%, and this can get as low as 10% throughout the free spectral range of the device. The ChemIcon instrument overcame this fault by brute force, using a much higher power laser than the more common instruments to generate the Raman scatter in experiments. In addition, the CCD itself was back thinned and had a high QE in the wavelength range of the experiment (and binning the CCD helps too - but at a loss of spatial resolution).
Tim Prusnick, PRUSNICK_TIM-at-worldnet.att.net Raman Application & Support Engineer Renishaw Inc. 623 Cooper Court Schaumburg, IL 60173 PHONE: 847-843-3666 FAX: 847-843-1744
-----Original Message----- } From: Jim Haley [mailto:haley-at-i-cubeinc.com] Sent: Monday, September 14, 1998 1:20 PM To: prusnick_tim-at-worldnet.att.net Cc: microscopy-at-Sparc5.Microscopy.Com
Timothy,
I'm curious about using a LCTF to filter the laser. All the LCTFs I have seen have a very poor transmissivity. Doesn't this make a large impact on the spectra mwhich is collected by the CCD?
Timothy M. Prusnick wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } The ChemIcon instrument uses the LCTF as a tunable bandpass filter. The } idea behind the instrument is to globally illuminate the sample with a large } laser spot and then collect all of the Raman scatter from the sample and } filter it through the LCTF. } } You then can collect spectra by taking a series of images with the CCD, each } at a different wavelength. If you select the same pixel in each of the } wavelength dependent images and plot it's intensity as a function of the } image wavelength, you can reproduce a spectra. Therefore, if you have a 100 } pixel by 100 pixel image, you can have 10000 spectra in one spectral image } dataset. The spectral resolution is based on the bandpass of the filter, } which is about 7 cm-1. The spatial resolution, as Fernando said, is } diffraction limited. } } Although I no longer work for the compant, I helped design the ChemIcon } instrument. So, if you have any questions about it's use or the science } behind it's design, please feel free to contact me. } } Tim Prusnick, PRUSNICK_TIM-at-worldnet.att.net } Raman Application & Support Engineer } Renishaw Inc. } 623 Cooper Court } Schaumburg, IL 60173 } PHONE: 847-843-3666 } FAX: 847-843-1744 } } -----Original Message----- } } From: Fernando Agullo-Rueda [mailto:FAR-at-icmm.csic.es] } Sent: Monday, September 14, 1998 2:35 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Raman imaging } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Kalpana S Katti wrote: } } } Does anayone on this network have experience with the new field of Raman } } Imaging ? We have a raman instrument with a tunable filter and imaging. I } } have a couple of questions. I am a TEM person and new at this so excuse } } me if these questions are too elementary. } } } } The manufacturer claims that on aquiring the image the spatial resolution } } of the 'spectra' is as small as a pixel. But since unlike IR, Raman is a } } scattering process wouldn't all regions limited by the beam (probe) , } } about 150 microns, be affected by each other and thus negating the claim } } of 1 pixel resolution? Infact I did observe this experimentaly while } } imaging composite polymeric samples. Spectra from all regions illuminated } } by the beam are almost identical although the image } } does exhibit contrast due to different phases. } } This question can be best answered by the manufacturer of } your instrument. However, I guess that your instrument has been } designed to get the best possible resolution. In that case, } as in other optical microscopy, the resolution is limited } by the diffraction of light. In other words, due to the wave nature } of light you can not focus a light beam below a certain size. } In the visible range that means that your spatial resolution } is around one micrometer on the sample. } } Probably the manufacturer has arranged things in such a way that } one micron on the sample is imaged at least as one pixel on the } CCD detector. } } I hope this helps. } } -------------------------------------------------------------------------- - } Fernando Agullo'-Rueda } Raman Microscopy Laboratory } Instituto de Ciencia de Materiales de Madrid (CSIC) } Cantoblanco, E-28049 Madrid } Espan~a (Spain) } } Tel: +34-91-334-9015 E-mail: {FAR-at-icmm.csic.es} } Fax: +34-91-372-0623 {http://www.icmm.csic.es/} } {http://www.icmm.csic.es/Fagullo/Fagullo.htm} } -------------------------------------------------------------------------- -
-- ****************************** Jim Haley Applications Engineer I-CUBE 2411 Crofton Lane, Suite 14A Crofton, MD 21114 voice: (301) 858-0505 fax: (301) 858-0615 web site: http://www.i-cubeinc.com e-mail: haley-at-i-cubeinc.com ******************************
A solution of 10g UA in 15mls H2O was measured with a Geiger counter. } 500 counts/sec was generated. A supplier had measured 100g UA :- 1 Alpha - {2 counts/sec, using a 540 scintillation meter with AP-2 Probe 2 Beta - } 500 counts/sec, using a 540 E1 probe coupled to a GM Meter (this determines beta events and some low energy gamma events) 3 Gamma dose Rate (energy field) - two measurements done: using Mini monitor tpye R with GM Probe - 0.6mR/hr (mainly gamma) and Ionisation chamber DMM 95/0500 - 5 mR/hr (Beta and Gamma energy field). 4 Specific Activity (U approx. 55%) = 1.04 x 10 { {...} } Bq { {...} } gm { {...} } .
Can UA be used openly without protection in laboratory?
Hi Group, =20 I am looking for anybody who does or is interested in photothermal = microscopy studies. I have applied photothermal techniques for optical microscopy = to enhance optical sensitivity during investigation of living cells. = However I have found very little info about the subject and I would like to exchange = with ideas with somebody who has similiar experience/interest. =20 Dmitri Lapotko =20 Luikov Heat and Mass Transfer Institute 15 Brovka Street Minsk Belarus =20 tel:(375172)842483 fax:(375172)842486 e-mail: ld-at-ns1.hmti.ac.by
} With my tongue only partway in my cheek, I ask... Alconox is a *mild* } detergent? What do you consider to be a harsh one? } I know no detergent is going to attack stainless, but read the fine } print on the packages before you try soaking on aluminum & its alloys, } some are alkaline enough to etch the surface. } Dear Robert, I'm doing the experiment. I mixed up a 1% solution of Alconox per the directions on the package. The pH is 9.1, so this is not too alkaline a detergent. At present, there is a piece of Al foil in the solution; I'll let you & the list know what fate the foil suffers. Yours, Bill Tivol
Hi everyone! My computer has been upgraded. My E-mail address has been changed to reflect my married name. New address is : brygv-at-ferro.com thanks, Vicky
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } Has anyone experienced puckering of tissues embedded in LRWhite resin? My tissues are cell cultures fixed in 2%PFA, .5%GA, dehydrated to75% ETOH then into graded LRWhite resin. I cut on a diamond knife and embed on 200 mesh grids. Before staining, the puckers and wrinkles are seen as small, and frequen, folds over most cells or along the cell borders. The resin areas are totally free of folding. It's as if the tissue area is picking up H2O during sectioning then has no where to go when it re-dries....very frustrating. After staining it's worse, as the stain seems to stay in the folds and gets really dark. Any thoughts and suggestions are very welcome as always. } Linda Fox } Loyola University } Stritch School of Medicine } lfox1-at-wpo.it.luc.edu } } Hi, You have several problems here. First, acrylics like LR White, etc., do not bond WITH the tissue as do epoxies. This allows a lot of shifting as soon as the stresses are relieved by cutting a thin section.. LR White also is modestly poorly crosslinked (as compared to the epoxides). You can increase the crosslinkage chemically (unless you are doing immunostaining, then you do not want to heavily crosslink). What to do? Do you need to use LR White? What is your purpose of using it. It has a number of downsides which one can trade off in the immunoprocessing protocols for better location of antigens. For standard TEM work is is inferior to epoxides (wrinkling of thick sections, poor crosslinkage, polymerization damage, difficulty sectioning because of its property of attracting water to the block face, etc.) What to do? If you must use LR White, try using a 100 mesh grid with a film. Also when picking up floating grids, DO NOT suck the water off under the grid with filter paper. Do draw the water off by sticking filter paper between the forceps. Then, lay the grid in the forcepts down to dry. Pull a lamp with a 60W bulb down over the forceps to slightly warm the situation. Use at least 5 forceps at once. That way some grids are drying and others are being picked up. This is the recommendation of Newman who holds the patent for LR White and LR Gold (He sold the license to the London Resin Company.) Should you decide to more heavily crosslink the resin, contact EMS for advice and the chemical. (I have no stock in EMS). If your sections have folds and you posstain them with heavy metals, the stain will accumulate in the folds. There is nothing you can do about that. You must avoid the wrinkles in the first place. Should you have more trouble, contact me. Bye, Hildy
An addendum to Melvyn Dickson's message, particularly for our younger readers:
When I started here at Reading, there was still in operation an EM6 microscope (AEI) which used a lot of what we Britishers call "valves" and Americans call "tubes" (what about Australians?). These all had an iridescent shine from the little bit of metal (magnesium?) which has been fired to scavenge the last traces of gas, a process known as GETTERING.
} Our coater is the Xenosput, obtained through Edwards. It uses xenon as the } sputtering gas. One (small) cylinder lasts ~ 10 years. The best part of } its operation is that the final purging of gases in the chamber is achieved } by actually sputtering titanium in the chamber itself so you only have } xenon left for the final coating stage.
Concerning our Korean friend's enquiry about phase contrast, I was at a Scanning Force Microscopy workshop in Bristol last week. Prof. Andrew Keller gave a most interesting historical talk, suggesting that the new technique would follow a three stage development similar to that of Electron Microscopy:
(1) ELATION: The EM allowed one to visualize a dimension previously only inferred from colloid studies;
(2) ANTICLIMAX: it looked at the surface only, and was prone to artefacts;
(3) STEADY PROVEN USEFULNESS.
Hence the importance of backing up with other techniques, particularly optical microscopy. As regards polymers, what pulled EM out of the doldrums in the late '50s was being able to see the same polymer crystals (solution grown polyethylene lamellae) both under the TEM and the newly available Zernicke Phase Contrast microscopes.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
ELECTRON MICROSCOPY RESEARCH TECHNICIAN POSITION AVAILABLE
Laboratory of William Lehman Ph. D. Department of Physiology Boston University School of Medicine 80 East Concord Street Boston, MA 02118
Qualifications:
1. Experience in molecular electron microscopy, particularly cryo-EM. 2. Facility with computer based-image analysis. 3. Familiarity with handling purified protein samples. 4. Ability to work independently in a small group environment.
Brief description of project:
We carry out state of the art - high resolution electron microscopy, computer assisted image analysis and three-dimensional image reconstruction to determine the arrangement of muscle thin filament components on F-actin and evaluate their position and influence on actin domains. Many significant projects, both on smooth and skeletal muscle systems, are being performed, and there is an excellent opportunity for a motivated individual to contribute to our understanding of muscle contractility and regulation.
Some recent publications:
Lehman, W., R. Craig & P. Vibert (1994) Ca-induced tropomyosin movement in Limulus thin filaments revealed by three-dimensional reconstruction. Nature 368, 65-67.
Lehman, W., P. Vibert, P. Uman & R. Craig (1995) Steric-blocking by tropomyosin visualized in relaxed vertebrate muscle thin filaments. J. Mol. Biol. 252, 191-196.
Vibert, P., R. Craig & W. Lehman (1997) Steric-model for activation of muscle thin filaments. J. Mol. Biol. 266, 8-14.
Hodgkinson, J.L., M. EL-Mezgueldi, R. Craig, P. Vibert, S.B. Marston & W. Lehman (1997) 3-D Image reconstruction of reconstituted smooth muscle thin filaments containing calponin: Visualization of interactions between F-actin and calponin. J. Mol. Biol. 273, 150-159.
Lehman, W., P. Vibert & R. Craig (1997) Visualization of caldesmon on smooth muscle thin filaments. J. Mol. Biol. 274, 310-317.
Hanein, D., N. Volkmann, Goldsmith, S., A.-M. Michon, W. Lehman, R. Craig, D. DeRosier, S. Almo & P. Matsudaira (1998) An atomic model of fimbrin binding to F-actin and its implications for filament crosslinking and regulation. Nature Struct. Biol. 5 787-792.
Send resume and 3 references to Dr. Lehman at above address or either FAX to (617)638-4273 or e-mail to lehman-at-med-rana.bu.edu.
We ion beam sputter Pt for polymer FESEM work up to ~X100,000; Pt gives us no significant coating artifacts up to that magnification and, unlike Cr, doesn't require pre-sputtering the target to remove the native oxide. For higher mags we will Cr coat.
Ev Osten 3M Company St. Paul, MN efosten-at-mmm.com
Stephen McCartney {stmccart-at-vt.edu} on 09/14/98 10:29:47 AM
To: Microscopy-at-sparc5.microscopy.com cc: (bcc: Ev Osten/US-Corporate/3M/US)
I am interested in high resolution coaters to use with field emission SEM. Does anyone working with FE-SEM's have some opinions for me on what is best specifically for polymer applications. Steve
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
HELP! I am new to the microscopy listserver and I am currently conducting undergraduate research for the Department of Biomedical Sciences at Southwest Missouri State University, located in Springfield, Missouri (USA). I am attempting to develop a TEM protocol to study the gap junction protein, Connexin-43 (Cx43) at the ultrastructural level. I am specifically interested in a protocol that utilizes colloidal gold as a marker and a post embedding technique that uses conventional resins. If anyone would be willing to share a specimen protocol for the localization of Cx43 or any other Connexins, please reply to me off list at: aaronrea-at-hotmail.com Thank you very much for your time!
Aaron Rea Department of Biomedical Sciences Southwest Missouri State University aaronrea-at-hotmail.com
I have some experience in evaporating silver, (2 - 3mm shot) from a tungsten wire basket, however, I am now faced with challenge of evaporating Au wire onto a similar substrate. Is it appropriate to simply wrap the Au wire around the larger diameter tungsten wire and proceed or is there a better approach. Your comments/suggestions are appreciated. Thanks.
Regards, Paul Gerroir Xerox Research Center of Canada
The Microscopy and Microanalysis Center (MMC) at the University of = Maryland at College Park is searching for a laboratory manager to = maintain and run two TEMs (Hitachi 600 AB, and JEOL 4000FX), one = electron microprobe (JEOL 8900), an environmental SEM (Electroscan E3) = and AFM (Dimension 3000) among other equipment. The MMC is a campus = facility that provides service to faculty, students, and outside users. = The facility is also used for teaching and research. The laboratory = manager would supervise two graduate students in the use of the = equipment.
Qualifications: The qualified candidate should have experience in the = maintenance of electron microscopes and their use. Background on = electronics and vacuum technology is required. Bachelor or Masters = degree in Materials Science, Physics or Engineering is preferred. Good = oral and written skills, as well as supervisory skills are required. =20
Salary: The starting salary is $30,000 to $35,000 depending on = experience. =20
Availability: October 1, 1998.
For best consideration, interested candidates should send curriculum = vitae and a list of three references to:=20 Dr. Lourdes Salamanca-Riba at either riba-at-eng.umd.edu,=20 Fax No. (301) 314-9467, or=20 Materials and Nuclear Engineering Department University of Maryland College Park, MD 20742-2115
The University of Maryland is an equal opportunity affirmative action = employer.
Are any of you interested in the possibility of using your micrographs to illustrate children's books? Here's a conference that should be quite informative:
Marine Biological Laboratory Hosts Institute for Children's Book Authors and Illustrators October 9-11, 1998
Woods Hole, MA-The Marine Biological Laboratory's Science Writing Fellowships Program and the Center for Children's Environmental Literature is co-sponsoring an Author, Illustrator, Biologist Institute. Working and aspiring children's book authors and illustrators, as well as scientists, are invited to participate in the three-day meeting. Organizers hope to foster new collaborations between authors, illustrators, and biologists.
For more information, contact: Pamela Clapp Hinkle Director of Communications Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 Tel: 508-289-7276 Fax: 508-457-1924 e-mail: pclapp-at-mbl.edu
For a complete program, see http://www.mbl.edu/html/MISC/AIB.html.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
In a message dated 98-09-15 19:07:25 EDT, you write: { { I have some experience in evaporating silver, (2 - 3mm shot) from a tungsten wire basket, however, I am now faced with challenge of evaporating Au wire onto a similar substrate. Is it appropriate to simply wrap the Au wire around the larger diameter tungsten wire and proceed or is there a better approach. Your comments/suggestions are appreciated. Thanks.
Regards, Paul Gerroir Xerox Research Center of Canada } }
Yes - it works well to wrap the wire around the tungsten filament.
A word of caution on the evaporation: Heat up the filament slowly, observing the gold through suitable dark glasses. At some point the gold will melt and run into a ball in the "v" of the filament. Then you can turn up the filament current until the gold evaporates.
Hello! I work with Atriplex nummularia Lidl. and this specie has a profusion of vesicular trichomes on both surfaces that are very fragile. I'm trying to obtain a transversal view of the leaf lamina to measure the thickness of the trichome layer.=20 Who has any idea? Thanks in advance. Rejane
Rejane Magalh=E3es Pimentel Galindo =20 ggalindo-at-elogica.com.br Universidade Federal Rural de Pernambuco Av. Boa Viagem, 6592/602 FAX: 55 (081) 4416177 51130-000, Recife, Pernambuco, Brasil
At 03:57 PM 9/15/98 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Paul,
I once watched someone hang a piece of wire from a larger tungsten wire, letting it dangle by a loop (like a clothes hangar). While every else predicted that the metal would simply fall off the wire when the loop heated up, the technician simply increased the current until the wire melted, forming a perfect adhering drop. He then increased the current until the metal evaporated and got a beautiful coating on his substrate. I don't remember if it was Au, Ag, or Pt, but the technique was simple, and it worked for him. Until then, we had been carefully wrapping the wire to evaporate around the supporting wire, like an electrical coil---a real skill for some of us.
For what it's worth.
Randy
} } Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
The concentration quoted is far higher (btw at what concentration in water does UA become a saturated solution?) than normally would be used for EM staining.
} A solution of 10g UA in 15mls H2O was measured with a Geiger counter.
I have been using UA for many years and I recall that whenever I questioned and investigated its possible radiation implications I have been assured that it is not dangerous at the concentrations and quantities we use, provided that it is not ingested. Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
I can second what Randy says. I have been evaporating gold wire off larger diameter tungsten wire (occasionally) for 30 years. It works, in an old Edwards coating unit. I must admit, I have always wrapped the gold around the tungsten, using two pairs of clean forceps.
Ann Fook Yang EM Unit Eastern Cereal and Oilseed Research Centre Agriculture and Agri-Food Canada 960 Carling Ave Central Experimental Farm Ottawa, Ontario Canada K1A 0C6
The concentration quoted is far higher (btw at what concentration in water does UA become a saturated solution?) than normally would be used for EM staining.
} A solution of 10g UA in 15mls H2O was measured with a Geiger counter.
I have been using UA for many years and I recall that whenever I questioned and investigated its possible radiation implications I have been assured that it is not dangerous at the concentrations and quantities we use, provided that it is not ingested. Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
I used to evaporate Au in a tungsten basket in an old Denton evap coater. We simply took a length of Au wire, wadded it up into a tiny ball and placed that in the basket. Took no large amount of manual dexterity and gave good results.
Gerroir, Paul J wrote:
} Hello Interested Readers, } } I have some experience in evaporating silver, (2 - 3mm shot) from a } tungsten wire basket, however, I am now faced with challenge of evaporating } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } wire around the larger diameter tungsten wire and proceed or is there a } better approach. Your comments/suggestions are appreciated. Thanks. } } Regards, } Paul Gerroir } Xerox Research Center of Canada
Dear Josephine, } } A solution of 10g UA in 15mls H2O was measured with a Geiger counter. } 500 } counts/sec was generated. } A supplier had measured 100g UA :- } 1 Alpha - {2 counts/sec, using a 540 scintillation meter with AP-2 Probe } 2 Beta - } 500 counts/sec, using a 540 E1 probe coupled to a GM Meter (this } determines beta events and some low energy gamma events) } 3 Gamma dose Rate (energy field) - two measurements done: } using Mini monitor tpye R with GM Probe - 0.6mR/hr (mainly gamma) } and Ionisation chamber DMM 95/0500 - 5 mR/hr (Beta and Gamma energy } field). } 4 Specific Activity (U approx. 55%) = 1.04 x 10 { {...} } Bq { {...} } gm } { {...} } . } I calculated approximately the expected activity from 30 g UA (about 0.1 mole, or 6*10^22 atoms). T_1/2 is 4.4*10^9 y and there are 3.1*10^7 s/y, so the decay rate is 5*10^-18 s^-1, and the activity is 3*10^5 Bq. The build-up of daughter products with shorter half-lives will reach steady state at which point the activities of the daughters will be the same as that of the parent. Pa 234 has a gamma transition, and there are several betas in the chain. The longer-lived isotopes in the chain have lives of 10^4 to 10^5 y, and these will not be at steady state (unless your UA is *very* old ;-) ). 0.6 mR/hr is a significant amount of exposure, and, if one were to hold the jars for some minutes, a sizable fraction of the allowed annual dose would be attained.
} Can UA be used openly without protection in laboratory? } Small amounts can be used, but be sure to wash hands before eating. One area of the lab should be used for UA. A quiet area with little traffic is best. UA, while not nearly the most dangerous EM reagent, should still be treated with respect. Yours, Bill Tivol
Responding to the message of {01J1VEY9MEFC8WXL78-at-pt.Cyanamid.COM} from "corwinl-at-pt.cyanamid.com"-at-Sparc5.Microscopy.Com: }
} According to the Handbook of Chemistry and Physics, the solubility of } UO2(C2H3O2)2.2H2O is 7.694 g/100 mL in 15 deg C water. } } } Leonard Corwin } Fort Dodge Animal Health } Princeton, NJ 08543-0400
That's just the information I was looking for!
In thinking about my own experiences with solubility of UA, I've noticed that even at only 2, 3, 4 % w/v, there always seems to be a bit that doesn't go into solution, even with heating. I consulted my vendor of UA and was told that there is an insoluble component present in commercial UA, and thats what I was seeing. This insoluble contaminant would then confuse efforts to determine saturation point by visual obvservation - are those insoluble contaminants or are they leftover UA crystals, having reached saturation? The yellow color of the solution foils any attempt to detect differences in color of the two kinds of crystals.
I did isolate these unknown crystals (forgot what color they were), did EDS analysis on 10 clumps of crystals and found that in addition to moderate to high amounts of U (quick, on-the-fly subjective semi-quant intuition), there were also high amounts of titanium (one very high), moderate to high amounts of silicon (one very high), low amount of aluminum (not always present), low amount of iron (always present), varying carbon and oxygen, low phosphorus. One cubic crystal was quite high in Si and Ti, only.
As U has one of the highest backscatter coefficients, perhaps the Ti, Fe, Al is coming from stray x-rays generated inside the SEM chamber by BSE's. If not, then my results indicate the basic composition of the contaminant crystals. Any idea what they might be?
I should now take UA crystals out of the bottle, measure their spectrum, to compare with the "contaminants" spectra measured above to sort this out.
But thanks to the above solubility data, we can at least make saturated solution of UA, and ignore or filter out the contaminant crystals.
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Well, you've probably heard enough ideas on this subject. But I'll give you my $0.02 worth (that's Au standard :-).)
Working as a lab technician at UW Astrophysics Lab in Madison, WI, I operated a vacuum deposition apparatus invented by a grad student which vaporized gold wire and deposited it on the surface of insulated copper wire to form the anode of a proportional detector. Anyway, in this apparatus the gold wire was simply wrapped around tungsten (heater filament) wire by hand. In vacuum, the tungsten filament was heated and the gold wire melted, resulting in gold vapor being deposited evenly on the wire rotissing nearby in the apparatus. It all seemed pretty crude to me, but worked just fine. And I didn't take any special precautions in wrapping the gold wire around the tungsten heater wire.
Regs, -- Tim ---
Gerroir, Paul J wrote: } Hello Interested Readers, } } I have some experience in evaporating silver, (2 - 3mm shot) from a } tungsten wire basket, however, I am now faced with challenge of evaporating } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } wire around the larger diameter tungsten wire and proceed or is there a } better approach. Your comments/suggestions are appreciated. Thanks. } } Regards, } Paul Gerroir } Xerox Research Center of Canada
-- ...we now return control of your computer screen to you... ------------------------------------------------------------ Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments, Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ------------------------------------------------------------ "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ------------------------------------------------------------
} EDX folks: } } Does anyone know of or plan to pick up support and/or development for } NIST's Desktop Spectrum Analyzer (DTSA) software?
At a DTSA workshop a couple of years past, Bob Myklebust *hinted* that he might start a private consulting business after retiring from NIST that would, among other things, provide DTSA training and support. But I haven't heard any announcements.
Now that DTSA has been "open sourced" by NIST, perhaps we need to start up a mailing list for user support. ( I'm CC-ing this to the microprobe list, which might be a more appropriate place for DTSA discussions until such a list can be set up. )
As long as you brought up this topic: Can anyone explain what exactly were the circumstances behind NIST's decision ? I got one semi-official explaination, which didn't exactly jive with either the currently posted comments on NIST's web pages nor with various rumours about law-suits that I heard from various vendors. Anyone know the story?
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |---
"I'm not as big a fool as I used to be, I'm a smaller fool." - Jack Kerouac Some of the Dharma {http://members.aol.com/kerouacsis/SomeDharma.html}
If you have Zephyr (model ZEM 300SW) water chillers and have experienced cooling/heating problems, please let me know how the problems were solved. One Zephyr won't cool-- water temp is 25 C. The other won't heat--temp is 8 C. Our A/C people have always been able to solve Haskris and Neslab chiller problems but are unable to solve Zephyr problems. Suggestions?
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
The Midwest Microscopy and Microanalysis Society will host a Materials Sciences meeting on October 9, 1998, at Purdue University. The meeting= has been organized by Eric Kvam and includes the following sessions:
9:00 Welcome and Introduction 9:20 Advances in Scanning Force Microscopy, R.P. Andres 10:00 High Resolution EELS on Superconductor Grain Boundaries, J.L. Lee=
10:20 Coffee Break 10:30 ESEM In situ Observation of Fatigue Crack Propagation, B.M. Hillb= erry 11:10 Electron Beam Lithography using the SEM, D.B. Janes 11:30 Lunch 1:00 Laboratories Tour 2:30 People to People: Electron Microscopy in China, J.L. Lee 2:50 Coffee Break 3:10 TEM of Electronic Composite Films, D.E. Collins 3:30 Advanced Image Analysis of Concrete Microstructures, S. Diamond 3:50 Dislocation Structure and Electronics in Semiconductors, V. Gopal
Pre-registration is NOT required, and there is no registration fee for members of MMMS. For non-members who wish to attend, it will be possi= ble to join the society at the meeting. Dues are $10 for regular members and = $5 for student members. Current MMMS members will receive a complete agenda a= nd travel information in the mail next week. Others may contact me via e-= mail at jane.a.fagerland-at-abbott.com, and meeting information will be mailed = or faxed to you.
Jane A. Fagerland, Ph.D MMMS Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064 (847) 935-0104 =
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Hi, Paul.
Well, you've probably heard enough ideas on this subject. But I'll give you my $0.02 worth (that's Au standard :-).)
Working as a lab technician at UW Astrophysics Lab in Madison, WI, I operated a vacuum deposition apparatus invented by a grad student which vaporized gold wire and deposited it on the surface of insulated copper wire to form the anode of a proportional detector. Anyway, in this apparatus the gold wire was simply wrapped around tungsten (heater filament) wire by hand. In vacuum, the tungsten filament was heated and the gold wire melted, resulting in gold vapor being deposited evenly on the wire rotissing nearby in the apparatus. It all seemed pretty crude to me, but worked just fine. And I didn't take any special precautions in wrapping the gold wire around the tungsten heater wire.
Regs, -- Tim ---
Gerroir, Paul J wrote: } Hello Interested Readers, } } I have some experience in evaporating silver, (2 - 3mm shot) from a } tungsten wire basket, however, I am now faced with challenge of evaporating } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } wire around the larger diameter tungsten wire and proceed or is there a } better approach. Your comments/suggestions are appreciated. Thanks. } } Regards, } Paul Gerroir } Xerox Research Center of Canada
-- ...we now return control of your computer screen to you... ------------------------------------------------------------ Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments, Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ------------------------------------------------------------ "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ------------------------------------------------------------
I would like to fix small spiders for TEM. I am not particular interested in preservation of specific structures, but would like to preserve both integumental and inner structures in general. I noticed that my specimens floated, and I am afraid that the fixation will not penetrate properly.
Does anybody has experience with fixation of similar specimens ? I would appreciate your comments very much.
Thank you.
Regards
Peter Funch
______________________________________________________________________ Peter Funch Assistant Professor, Ph.D.
Department of Zoology Direct Line + 45 8942 2764 Institute of Biological Sciences Secretary + 45 8942 2727 University of Aarhus Telefax + 45 86 12 51 75 Universitetsparken E-mail: peter.funch-at-biology.aau.dk Building 135 DK-8000 Aarhus C Denmark ______________________________________________________________________
Department of Cell and Structural Biology University of Illinois at Urbana Champaign ---------------------------------------------------------------------------
Qualifications: Minimum requirement: Bachelor of Science degree. Applicants with advance degrees are invited to apply. Previous lab and/or electron microscopy experience required.
Responsibilities: To perform research in a laboratory of cell biology and structural biology. Work will combine electron microscopy and light microscopy with tissue culture, immunostaining, and molecular biology.
Salary: Dependent on qualifications.
Starting Date: As soon as possible after closing date.
Send Applications to: Ms. Helen Neef (asb) Department of Cell and Structural Biology B107 Chemical and Life Sciences Laboratory University of Illinois 601 S. Goodwin Avenue Urbana, IL 61801 Phone: (217) 244-6638
Closing Date: Nov. 15, 1998. Interviews may be conducted before the closing date, but all applications received by that date will receive full consideration and the final selection will not be made until after that date.
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN IS AN AFFIRMATIVE ACTION, EQUAL OPPORTUNITY EMPLOYER
****************************************************** Andrew Belmont 217-244-1648 (fax) Associate Professor 217-244-2311 (office) Department of Cell and Structural Biology asbel-at-uiuc.edu University of Illinois, Urbana-Champaign B107, 601 S. Goodwin Ave. Urbana, IL 61801 ******************************************************
Hi, I have a request for a micrograph of the pits and lands on a music CD. I thought this would be fun and simple but we aren't having any luck imaging anything (actually it was so reflective we imaged the detector!:-) Here's what we've done: We cut up a CD - using a piece of it close to the center hole (so we knew it had something on it to see). The piece was mounted (lower surface up) on a stub using a carbon sticky tab and silver paste. The sample was sputter coated for 2 minutes (some for 3) and scoped. The first time we viewed it we didn't coat it (see paragraph one for details).
Do we need a solvent to munch awhile on the surface layer of the CD? Any help would be greatly appreciated!
It was a Christmas music CD so maybe that is the problem :-).
beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
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Hi, Paul.
Well, you've probably heard enough ideas on this subject. But I'll give you my $0.02 worth (that's Au standard :-).)
Working as a lab technician at UW Astrophysics Lab in Madison, WI, I operated a vacuum deposition apparatus invented by a grad student which vaporized gold wire and deposited it on the surface of insulated copper wire to form the anode of a proportional detector. Anyway, in this apparatus the gold wire was simply wrapped around tungsten (heater filament) wire by hand. In vacuum, the tungsten filament was heated and the gold wire melted, resulting in gold vapor being deposited evenly on the wire rotissing nearby in the apparatus. It all seemed pretty crude to me, but worked just fine. And I didn't take any special precautions in wrapping the gold wire around the tungsten heater wire.
Regs, -- Tim ---
Gerroir, Paul J wrote: } Hello Interested Readers, } } I have some experience in evaporating silver, (2 - 3mm shot) from a } tungsten wire basket, however, I am now faced with challenge of evaporating } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } wire around the larger diameter tungsten wire and proceed or is there a } better approach. Your comments/suggestions are appreciated. Thanks. } } Regards, } Paul Gerroir } Xerox Research Center of Canada
-- ...we now return control of your computer screen to you... ------------------------------------------------------------ Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments, Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ------------------------------------------------------------ "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ------------------------------------------------------------
Numbers of JEOL owners in Oz have complained of similar oil contamination. When we last evaluated FESEMs the JEOL rep was persistent to know why we didn't like the JEOL models. One of my reasons was the oil contamination problem and I asked for some explanation as to why it was so common.
His answer was that users were opening the inner door of the airlock too soon and perhaps too quickly. There was too much air left in the airlock and the pressure rise in the chamber stalled the diffusion pump and backstreaming of the diffusion pump oil caused the contamination.
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Hi all, I was just wondering why everyone uses gold wire? Last time I looked, 5 9's gold splatters were the cheapest. With wire and shot you have to pay for the manufacturing/molding costs. I have always used a tungstun or moly boat, so there is no need to wrap or hang anything. -regards -andrew
--ListServer-at-MSA.Microscopy.Com
On Wed, 16 Sep 1998 10:02:47 Keith Ryan wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-----== Sent via Deja News, The Discussion Network ==----- http://www.dejanews.com/ Easy access to 50,000+ discussion forums
Hi all, I was just wondering why everyone uses gold wire? Last time I looked, 5 9's gold splatters were the cheapest. With wire and shot you have to pay for the manufacturing/molding costs. I have always used a tungstun or moly boat, so there is no need to wrap or hang anything. -regards -andrew
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On Wed, 16 Sep 1998 10:02:47 Keith Ryan wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-----== Sent via Deja News, The Discussion Network ==----- http://www.dejanews.com/ Easy access to 50,000+ discussion forums
Hi all, I was just wondering why everyone uses gold wire? Last time I looked, 5 9's gold splatters were the cheapest. With wire and shot you have to pay for the manufacturing/molding costs. I have always used a tungstun or moly boat, so there is no need to wrap or hang anything. -regards -andrew
--ListServer-at-MSA.Microscopy.Com
On Wed, 16 Sep 1998 10:02:47 Keith Ryan wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-----== Sent via Deja News, The Discussion Network ==----- http://www.dejanews.com/ Easy access to 50,000+ discussion forums
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Hi, Paul.
Well, you've probably heard enough ideas on this subject. But I'll give you my $0.02 worth (that's Au standard :-).)
Working as a lab technician at UW Astrophysics Lab in Madison, WI, I operated a vacuum deposition apparatus invented by a grad student which vaporized gold wire and deposited it on the surface of insulated copper wire to form the anode of a proportional detector. Anyway, in this apparatus the gold wire was simply wrapped around tungsten (heater filament) wire by hand. In vacuum, the tungsten filament was heated and the gold wire melted, resulting in gold vapor being deposited evenly on the wire rotissing nearby in the apparatus. It all seemed pretty crude to me, but worked just fine. And I didn't take any special precautions in wrapping the gold wire around the tungsten heater wire.
Regs, -- Tim ---
Gerroir, Paul J wrote: } Hello Interested Readers, } } I have some experience in evaporating silver, (2 - 3mm shot) from a } tungsten wire basket, however, I am now faced with challenge of evaporating } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } wire around the larger diameter tungsten wire and proceed or is there a } better approach. Your comments/suggestions are appreciated. Thanks. } } Regards, } Paul Gerroir } Xerox Research Center of Canada
-- ...we now return control of your computer screen to you... ------------------------------------------------------------ Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments, Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ------------------------------------------------------------ "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ------------------------------------------------------------
Hi all, I was just wondering why everyone uses gold wire? Last time I looked, 5 9's gold splatters were the cheapest. With wire and shot you have to pay for the manufacturing/molding costs. I have always used a tungstun or moly boat, so there is no need to wrap or hang anything. -regards -andrew
--ListServer-at-MSA.Microscopy.Com
On Wed, 16 Sep 1998 10:02:47 Keith Ryan wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-----== Sent via Deja News, The Discussion Network ==----- http://www.dejanews.com/ Easy access to 50,000+ discussion forums
Hi all, I was just wondering why everyone uses gold wire? Last time I looked, 5 9's gold splatters were the cheapest. With wire and shot you have to pay for the manufacturing/molding costs. I have always used a tungstun or moly boat, so there is no need to wrap or hang anything. -regards -andrew
--ListServer-at-MSA.Microscopy.Com
On Wed, 16 Sep 1998 10:02:47 Keith Ryan wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-----== Sent via Deja News, The Discussion Network ==----- http://www.dejanews.com/ Easy access to 50,000+ discussion forums
Hi Group, I am looking for anybody who does or is interested in photothermal microscopy studies. I have applied photothermal techniques for optical microscopy to enhance optical sensitivity during investigation of living cells. However I have found very little info about the subject and I would like to exchange with ideas with somebody who has similiar experience/interest. Dmitri Lapotko Luikov Heat and Mass Transfer Institute 15 Brovka Street Minsk Belarus tel:(375172)842483 fax:(375172)842486 e-mail: ld-at-ns1.hmti.ac.by
In a message dated 98-09-16 20:01:57 EDT, you write:
{ { Hi, I have a request for a micrograph of the pits and lands on a music CD. I thought this would be fun and simple but we aren't having any luck imaging anything (actually it was so reflective we imaged the detector!:-) Here's what we've done: We cut up a CD - using a piece of it close to the center hole (so we knew it had something on it to see). The piece was mounted (lower surface up) on a stub using a carbon sticky tab and silver paste. The sample was sputter coated for 2 minutes (some for 3) and scoped. The first time we viewed it we didn't coat it (see paragraph one for details).
Do we need a solvent to munch awhile on the surface layer of the CD? Any help would be greatly appreciated!
It was a Christmas music CD so maybe that is the problem :-).
beth } }
How about making a replica and imaging it in the TEM?
If you wish to do that I can give you some technique hints.
I have been asked to compare the grain size of 2 MgO aggregates. The samples were embedded and polished with diamond paste, but I am not happy with the result. The surface looks smeared. Does the sample need to be etched or do I need to evaluate my polishing technique???? Thanks for you help.
Nan Laudenslager Specialty Minerals, Inc. Easton, PA nhl-at-early.com
=46or Lorentz microscopy purposes we have modified a 200CX such that specime= n is located in a non-traditional position of the column. For the specimen to be in focus the objective lens is operating at a level that is 20% higher than normal. We would like to determine the Cs of the objective lens under these conditions. Note: the image resolution is about 50=C5 under these conditions. Any ideas??
Thanks,
Mark A. Wall L-350 7000 east Ave Chemistry & Materials Science Dept. Lawrence Livermore National Lab Livermore, CA 94550 925 423-7162
All the work we did in the past proved to us beyond doubt that the oil wa= s from the rotary pump. No matter how sophisticated the system is if you u= se a rotary pump at all in the cycle you do seem to get RP oil as a contaminant.
I think the guy had a good try but I for one do not believe him!
Hi all. Great topic. We have been involved in building a new control unit for a factory which = evaporates aluminium onto plastic parts to give them that silver look. This has given us a lot more understanding of the coating technique as = they have very large vacuum chambers and many samples that need an even = and smooth coat each time. What they do is to use tungsten wires coils that have two strands of W = wire twisted together as aposed to, what we all seem to use, the single = strand of W wire. The aluminium wire is then twisted by hand onto this = coil, fairly loosely but just that it makes contact. The reasoning is = that as you start the heating of the aluminium it will melt and flow, = via capillary action, between the two W wires. This means that the = aluminium is then in very good contact with the W coil and a lower = current is needed to vaporise the aluminium. You also have a source of = aluminium the whole length of the W coil each time. This ensures a even = and repeatable coat each time.
Cheers Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
-----Original Message----- } From: Timothy Moeller [SMTP:tmoeller-at-noran.com] Sent: Wednesday, September 16, 1998 7:15 PM To: Paul.Gerroir-at-crt.xerox.com Cc: Microscopy-at-sparc5.microscopy.com; Microscopy-at-sparc5.microscopy.com
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Hi, Paul.
Well, you've probably heard enough ideas on this subject. But I'll give you my $0.02 worth (that's Au standard :-).)
Working as a lab technician at UW Astrophysics Lab in Madison, WI, I operated a vacuum deposition apparatus invented by a grad student which vaporized gold wire and deposited it on the surface of insulated copper wire to form the anode of a proportional detector. Anyway, in this apparatus the gold wire was simply wrapped around tungsten (heater filament) wire by hand. In vacuum, the tungsten filament was heated and the gold wire melted, resulting in gold vapor being deposited evenly on the wire rotissing nearby in the apparatus. It all seemed pretty crude to me, but worked just fine. And I didn't take any special precautions in wrapping the gold wire around the tungsten heater wire.
Regs, -- Tim ---
Gerroir, Paul J wrote: } Hello Interested Readers, } =20 } I have some experience in evaporating silver, (2 - 3mm shot) from a } tungsten wire basket, however, I am now faced with challenge of = evaporating } Au wire onto a similar substrate. Is it appropriate to simply wrap the = Au } wire around the larger diameter tungsten wire and proceed or is there = a } better approach. Your comments/suggestions are appreciated. Thanks. } =20 } Regards, } Paul Gerroir } Xerox Research Center of Canada
--=20 ...we now return control of your computer screen to you... ------------------------------------------------------------ Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments, Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ------------------------------------------------------------ "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ------------------------------------------------------------
Jon, In the message below, I found a reference to a microprobe listserver. You should check it out. The website is at http://www.microanalysis.org/mas/maslistserver/maslistserver.html
This mailing list is sponsored by the Microbeam Analysis Society and managed by John Mansfield and Greg Meeker. More information is available at at the MAS web. To subscribe send email to the mailing list at:
microprobe-at-www.microanalysis.org with the word "subscribe" in the subject.
Hope this is as useful as the microscopy listserver is to me.
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On Wed, 16 Sep 1998, Heeschen, Bill (WA) wrote:
} EDX folks: } } Does anyone know of or plan to pick up support and/or development for } NIST's Desktop Spectrum Analyzer (DTSA) software?
At a DTSA workshop a couple of years past, Bob Myklebust *hinted* that he might start a private consulting business after retiring from NIST that would, among other things, provide DTSA training and support. But I haven't heard any announcements.
Now that DTSA has been "open sourced" by NIST, perhaps we need to start up a mailing list for user support. ( I'm CC-ing this to the microprobe list, which might be a more appropriate place for DTSA discussions until such a list can be set up. )
As long as you brought up this topic: Can anyone explain what exactly were the circumstances behind NIST's decision ? I got one semi-official explaination, which didn't exactly jive with either the currently posted comments on NIST's web pages nor with various rumours about law-suits that I heard from various vendors. Anyone know the story?
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |---
"I'm not as big a fool as I used to be, I'm a smaller fool." - Jack Kerouac Some of the Dharma {http://members.aol.com/kerouacsis/SomeDharma.html}
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Hi, Paul.
Well, you've probably heard enough ideas on this subject. But I'll give you my $0.02 worth (that's Au standard :-).)
Working as a lab technician at UW Astrophysics Lab in Madison, WI, I operated a vacuum deposition apparatus invented by a grad student which vaporized gold wire and deposited it on the surface of insulated copper wire to form the anode of a proportional detector. Anyway, in this apparatus the gold wire was simply wrapped around tungsten (heater filament) wire by hand. In vacuum, the tungsten filament was heated and the gold wire melted, resulting in gold vapor being deposited evenly on the wire rotissing nearby in the apparatus. It all seemed pretty crude to me, but worked just fine. And I didn't take any special precautions in wrapping the gold wire around the tungsten heater wire.
Regs, -- Tim ---
Gerroir, Paul J wrote: } Hello Interested Readers, } } I have some experience in evaporating silver, (2 - 3mm shot) from a } tungsten wire basket, however, I am now faced with challenge of evaporating } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } wire around the larger diameter tungsten wire and proceed or is there a } better approach. Your comments/suggestions are appreciated. Thanks. } } Regards, } Paul Gerroir } Xerox Research Center of Canada
-- ...we now return control of your computer screen to you... ------------------------------------------------------------ Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments, Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ------------------------------------------------------------ "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ------------------------------------------------------------
On 16 Sep 98 at 16:52, The slowly moving finger of Beth Richardson wrote:
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- } ---. } } } Hi, } I have a request for a micrograph of the pits and lands on a music } CD. I thought this would be fun and simple but we aren't having any } luck imaging anything (actually it was so reflective we imaged the } detector!:-) Here's what we've done: We cut up a CD - using a piece } of it close to the center hole (so we knew it had something on it to } see). The piece was mounted (lower surface up) on a stub using a } carbon sticky tab and silver paste. The sample was sputter coated } for 2 minutes (some for 3) and scoped. The first time we viewed it } we didn't coat it (see paragraph one for details). } } Do we need a solvent to munch awhile on the surface layer of the CD? } Any help would be greatly appreciated! } } It was a Christmas music CD so maybe that is the problem :-). } } beth } Beth I have done this in the past using a rewritable CD where the reflective coating can be readily stripped using tape. Using a normal CD I think we had to physically rip it apart, ( I seem to remember it was quite tough ), a good source for this is the freebies that you find on computer mags. John John Findlay Science Faculty EM Facility. Edinburgh University. Daniel Rutherford Bldg. Kings Buildings. Edinburgh EH9 3JH. tel. 0131-650-5344 fax. 0131-650-6563 John.Findlay-at-ed.ac.uk
this may be of no help what-so-ever, but we have a Flowcool water cooler. It has a digital readout of temperature and at present it 'thinks' that the water is about 70 deg C (ie temp readout is a bout 70) whereas the true temperature on the microscope is between 16 to 18 deg C.
This has happened once before and it turned out that it was the temperature sensor which was easy and fairly cheap to replace - so we did it and it worked. It looks like it's gone again (not very reliable) after 3 or 4 years and will be replaced soon.
You might just have a similar problem.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: John C. Wheatley To: Microscopy
If you have Zephyr (model ZEM 300SW) water chillers and have experienced cooling/heating problems, please let me know how the problems were solved. One Zephyr won't cool-- water temp is 25 C. The other won't heat--temp is 8 C. Our A/C people have always been able to solve Haskris and Neslab chiller problems but are unable to solve Zephyr problems. Suggestions?
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
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Hi, Paul.
Well, you've probably heard enough ideas on this subject. But I'll give you my $0.02 worth (that's Au standard :-).)
Working as a lab technician at UW Astrophysics Lab in Madison, WI, I operated a vacuum deposition apparatus invented by a grad student which vaporized gold wire and deposited it on the surface of insulated copper wire to form the anode of a proportional detector. Anyway, in this apparatus the gold wire was simply wrapped around tungsten (heater filament) wire by hand. In vacuum, the tungsten filament was heated and the gold wire melted, resulting in gold vapor being deposited evenly on the wire rotissing nearby in the apparatus. It all seemed pretty crude to me, but worked just fine. And I didn't take any special precautions in wrapping the gold wire around the tungsten heater wire.
Regs, -- Tim ---
Gerroir, Paul J wrote: } Hello Interested Readers, } } I have some experience in evaporating silver, (2 - 3mm shot) from a } tungsten wire basket, however, I am now faced with challenge of evaporating } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } wire around the larger diameter tungsten wire and proceed or is there a } better approach. Your comments/suggestions are appreciated. Thanks. } } Regards, } Paul Gerroir } Xerox Research Center of Canada
-- ...we now return control of your computer screen to you... ------------------------------------------------------------ Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments, Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ------------------------------------------------------------ "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ------------------------------------------------------------
The pits and lands on a CD are { {inside} } the plastic of the CD, so you need to dissolve it with some sort of solvent (I believe I used methanol, but can't recall for sure). The correct organic solvent will make the plastic disappear entirely, not just craze it and turn it cloudy. I think there may be several types of plastic used as the base, because I used toluene a number of years ago the first time I tried this, and it didn't work on the CD I used a few months ago. Anyway, use a couple of fresh changes until the foil is floating free, then mount it on a stub and take a look. You might need to mount both sides of a single piece so that you are sure you have the side with the pits. If you've gotten all the plastic off, you might get away without coating, but I coated mine anyway.
You can see an example of the pits and lands at http://www.mta.ca/~jehrman/cd.htm BTW, the CD I used is a Microsoft(TM) Office demo CD, so any flaws are Bill Gate's fault, not mine!
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------. } } Hi, } I have a request for a micrograph of the pits and lands on a music CD. } I } thought this would be fun and simple but we aren't having any luck } imaging } anything (actually it was so reflective we imaged the detector!:-) } Here's what we've done: } We cut up a CD - using a piece of it close to the center hole (so we } knew } it had something on it to see). The piece was mounted (lower surface } up) on } a stub using a carbon sticky tab and silver paste. The sample was } sputter } coated for 2 minutes (some for 3) and scoped. The first time we viewed } it } we didn't coat it (see paragraph one for details). } } Do we need a solvent to munch awhile on the surface layer of the CD? } Any help would be greatly appreciated! } } It was a Christmas music CD so maybe that is the problem :-). } } beth } } ************************************** } Beth Richardson } EM Lab Coordinator } Botany Department } University of Georgia } Athens, GA 30602 } } Phone - (706) 542-1790 } FAX - (706) 542-1805 } Email - beth-at-dogwood.botany.uga.edu } **************************************
There was a thread about CDs recently which might be helpful. In a mass produced CD the pits are in the aluminum layer which is under a plastic layer. Electron imaging won't stand a chance unles the plastic is removed. BTW... CD-Rs are quite different - use a dye rather than Al film.
Woody McDermott Technology
me: http://www.geocities.com/capecanaveral/3722 Please pardon the commercials!
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In a message dated 98-09-16 20:01:57 EDT, you write:
{ { Hi, I have a request for a micrograph of the pits and lands on a music CD. I thought this would be fun and simple but we aren't having any luck imaging anything (actually it was so reflective we imaged the detector!:-) Here's what we've done: We cut up a CD - using a piece of it close to the center hole (so we knew it had something on it to see). The piece was mounted (lower surface up) on
a stub using a carbon sticky tab and silver paste. The sample was sputter coated for 2 minutes (some for 3) and scoped. The first time we viewed it we didn't coat it (see paragraph one for details).
Do we need a solvent to munch awhile on the surface layer of the CD? Any help would be greatly appreciated!
It was a Christmas music CD so maybe that is the problem :-).
beth } }
How about making a replica and imaging it in the TEM?
If you wish to do that I can give you some technique hints.
Ted, I have good luck with CD images. Gold coat then tilt your sample as much as you can in your chamber, up to 85 degrees if possible. You should be able to see the pits and any defects such as shoulders around the pits, if present. I got a circular section (about 1.5 inch diameter) of the CD without introducing any stress by having it punched out in the machine shop. Hope this helps. ---------- } From: DUNNTEM-at-aol.com To: beth; Jackie Terry; Wayne England Cc: Microscopy -----------------------------------------------------------------------.
In a message dated 98-09-16 20:01:57 EDT, you write:
{ { Hi, I have a request for a micrograph of the pits and lands on a music CD. I thought this would be fun and simple but we aren't having any luck imaging anything (actually it was so reflective we imaged the detector!:-) Here's what we've done: We cut up a CD - using a piece of it close to the center hole (so we knew it had something on it to see). The piece was mounted (lower surface up) on a stub using a carbon sticky tab and silver paste. The sample was sputter coated for 2 minutes (some for 3) and scoped. The first time we viewed it we didn't coat it (see paragraph one for details).
Do we need a solvent to munch awhile on the surface layer of the CD? Any help would be greatly appreciated!
It was a Christmas music CD so maybe that is the problem :-).
beth } }
How about making a replica and imaging it in the TEM?
If you wish to do that I can give you some technique hints.
On Wed, 16 Sep 1998 DUNNTEM-at-aol.com-at-sparc5.microscopy.com wrote:
} I have a request for a micrograph of the pits and lands on a music CD. I } thought this would be fun and simple but we aren't having any luck imaging } anything (actually it was so reflective we imaged the detector!:-)
The pits and lands on the CD are on a metal layer that is sandwiched with a plastic one. The best way to image the metal surface is to delaminate the CD.
Nan Laudenslager wrote: ================================================ I have been asked to compare the grain size of 2 MgO aggregates. The samples were embedded and polished with diamond paste, but I am not happy with the result. The surface looks smeared. Does the sample need to be etched or do I need to evaluate my polishing technique???? ================================================= Certainly if there is room for improvement in the polishing technique, that is the first thing to do. However, when all else fails and you still have smearing (not unusual), then the smeared polymer can be selectively removed using a barrel type (isotropic) RF plasma etcher, using of course, oxygen. It is quite effective and it is a room temperature process. It takes only a few minutes of etching if that much.
Several commercially available table top etchers are available including our SPI Plasma Prep II, the details of which are on our website below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I seldom evaporate Au, but use W-wire baskets when I do... My source of Au is from "spent" sputter targets from years past. Why buy wire when I have a few perforated sputter targets around!
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I used to evaporate Au in a tungsten basket in an old Denton evap coater. We simply took a length of Au wire, wadded it up into a tiny ball and placed that in the basket. Took no large amount of manual dexterity and gave good results.
Gerroir, Paul J wrote:
} Hello Interested Readers, } } I have some experience in evaporating silver, (2 - 3mm shot) from a } tungsten wire basket, however, I am now faced with challenge of evaporating } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } wire around the larger diameter tungsten wire and proceed or is there a } better approach. Your comments/suggestions are appreciated. Thanks. } } Regards, } Paul Gerroir } Xerox Research Center of Canada
HELP! I am new to the microscopy listserver and I am currently conducting undergraduate research for the Department of Biomedical Sciences at Southwest Missouri State University, located in Springfield, Missouri (USA). I am attempting to develop a TEM protocol to study the gap junction protein, Connexin-43 (Cx43) at the ultrastructural level. I am specifically interested in a protocol that utilizes colloidal gold as a marker and a post embedding technique that uses conventional resins. If anyone would be willing to share a specimen protocol for the localization of Cx43 or any other Connexins, please reply to me off list at: aaronrea-at-hotmail.com Thank you very much for your time!
Aaron Rea Department of Biomedical Sciences Southwest Missouri State University aaronrea-at-hotmail.com
______________________________________________________ Get Your Private, Free Email at http://www.hotmail.com
The simplest way that was suggested by Brian McIntyre and what worked fine was to simply scratch the back surface with a knife and take some good scotch tape and press on the back over the scratch with the tape. When it has been pressed onto the back with a lot of pressure, simply peel it off. You'll see the pattern transferred to the tape. Since this layer is conductive, there is no need to coat it as long as you have a good conductive path from that surface to the support stub. You can use carbon paint.
If you have the conductive carbon double sticky pads, they work well also.
-Scott Walck Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: beth-at-dogwood.botany.uga.edu To: Microscopy-at-sparc5.microscopy.com
Hi, I have a request for a micrograph of the pits and lands on a music CD. I thought this would be fun and simple but we aren't having any luck imaging anything (actually it was so reflective we imaged the detector!:-) Here's what we've done: We cut up a CD - using a piece of it close to the center hole (so we knew it had something on it to see). The piece was mounted (lower surface up) on a stub using a carbon sticky tab and silver paste. The sample was sputter coated for 2 minutes (some for 3) and scoped. The first time we viewed it we didn't coat it (see paragraph one for details).
Do we need a solvent to munch awhile on the surface layer of the CD? Any help would be greatly appreciated!
It was a Christmas music CD so maybe that is the problem :-).
beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
We have several old American Optical/Spenser stereo microscopes that need a round glass stage insert. The inserts are 4" (10 cm) in diameter. I know we can have these made locally by a glass shop but was wondering whether anyone knows of an "off the shelf" source.
I am looking for the source of microparticles to colibrate/model light absorption processes. It can be latex spheres with diameter 1-10 um, partly stained with dyes(is it possible?). Any input will be appreciated.
Thanks
Dmitri Lapotko Luikov Heat and Mass Transfer Institute Minsk Belarus tel:(375172)842483 e-mail:ld-at-ns1.hmti.ac.by
Yes, having TWO (or more) tungsten filaments twisted together is important, to give the capillary action for more even dispersal as you mention. I forgot that important detail (it's been 27 years!) Thanks for jogging my memory. In fact, as I remember now, we were actually taking a long strand of tungsten wire and doubling it over more than once, to give a twisted set of 4 or more for our apparatus. We then wrapped the gold wire around that.
Regs, -- Tim
Luc Harmsen wrote: } Hi all. } Great topic. } We have been involved in building a new control unit for a factory which evaporates aluminium onto plastic parts to give them that silver look. } This has given us a lot more understanding of the coating technique as they have very large vacuum chambers and many samples that need an even and smooth coat each time. } What they do is to use tungsten wires coils that have two strands of W wire twisted together as aposed to, what we all seem to use, the single strand of W wire. The aluminium wire is then twisted by hand onto this coil, fairly loosely but just that it makes contact. The reasoning is that as you start the heating of the aluminium it will melt and flow, via capillary action, between the two W wires. This means that the aluminium is then in very good contact with the W coil and a lower current is needed to vaporise the aluminium. You also have a source of aluminium the whole length of the W coil each time. This ensures a even and repeatable coat each time. } } Cheers } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za } } -----Original Message----- } } From: Timothy Moeller [SMTP:tmoeller-at-noran.com] } Sent: Wednesday, September 16, 1998 7:15 PM } To: Paul.Gerroir-at-crt.xerox.com } Cc: Microscopy-at-sparc5.microscopy.com; Microscopy-at-sparc5.microscopy.com } Subject: Re: Evaporation of Gold } } Hi, Paul. } } Well, you've probably heard enough ideas on this subject. But I'll give } you my $0.02 worth (that's Au standard :-).) } } Working as a lab technician at UW Astrophysics Lab in Madison, WI, I } operated a vacuum deposition apparatus invented by a grad student which } vaporized gold wire and deposited it on the surface of insulated copper } wire to form the anode of a proportional detector. Anyway, in this } apparatus the gold wire was simply wrapped around tungsten (heater } filament) wire by hand. In vacuum, the tungsten filament was heated and } the gold wire melted, resulting in gold vapor being deposited evenly on } the wire rotissing nearby in the apparatus. It all seemed pretty crude } to me, but worked just fine. And I didn't take any special precautions } in wrapping the gold wire around the tungsten heater wire. } } Regs, } -- Tim } --- } } Gerroir, Paul J wrote: } } Hello Interested Readers, } } } } I have some experience in evaporating silver, (2 - 3mm shot) from a } } tungsten wire basket, however, I am now faced with challenge of evaporating } } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } } wire around the larger diameter tungsten wire and proceed or is there a } } better approach. Your comments/suggestions are appreciated. Thanks. } } } } Regards, } } Paul Gerroir } } Xerox Research Center of Canada }
-- ..we now return control of your computer screen to you... ---------------------------------------------------------- Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ---------------------------------------------------------- "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ----------------------------------------------------------
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Hi, Paul.
Well, you've probably heard enough ideas on this subject. But I'll give you my $0.02 worth (that's Au standard :-).)
Working as a lab technician at UW Astrophysics Lab in Madison, WI, I operated a vacuum deposition apparatus invented by a grad student which vaporized gold wire and deposited it on the surface of insulated copper wire to form the anode of a proportional detector. Anyway, in this apparatus the gold wire was simply wrapped around tungsten (heater filament) wire by hand. In vacuum, the tungsten filament was heated and the gold wire melted, resulting in gold vapor being deposited evenly on the wire rotissing nearby in the apparatus. It all seemed pretty crude to me, but worked just fine. And I didn't take any special precautions in wrapping the gold wire around the tungsten heater wire.
Regs, -- Tim ---
Gerroir, Paul J wrote: } Hello Interested Readers, } } I have some experience in evaporating silver, (2 - 3mm shot) from a } tungsten wire basket, however, I am now faced with challenge of evaporating } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } wire around the larger diameter tungsten wire and proceed or is there a } better approach. Your comments/suggestions are appreciated. Thanks. } } Regards, } Paul Gerroir } Xerox Research Center of Canada
-- ...we now return control of your computer screen to you... ------------------------------------------------------------ Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments, Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ------------------------------------------------------------ "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ------------------------------------------------------------
SCANNING PROBE MICROSCOPY: PRINCIPLES AND PRACTICE.
18-22 January 1999
Short Course from the School of Mechanical and Materials Engineering, University of Surrey, Guildford, UK.
THE COURSE
The aim of this five day intensive course is to introduce the principles and practice of Scanning Tunnelling Microscopy (STM), Scanning Force Microscopy (SFM) and other methods of Scanning Probe Microscopy (SPM). The physical concepts employed in the instrumentation of STM and SFM are simple, but the interpretation of the STM and SFM results can be complicated because of the convolution of several interactions in the measurement process. This complication exists in the large scale imaging of surface morphology as well as in the molecular- and atomic scale images. Thus, many STM and SFM studies can be misinterpreted. To help to alleviate this problem, we felt it necessary to bring together in this course the essential components of STM and SFM studies, namely, the practical aspects of STM and SFM, the image simulation and the qualitative evaluations of tip force induced surface corrugations.
The primary goal of the course will be to describe how the surfaces of various materials are characterised by employing STM, SFM and other methods of SPM and what physical/chemical features can be deducted from their images.
This will be achieved through a balance of lectures, tutorials and laboratory demonstrations. The course will provide a theoretical introduction to the field and an overview of recent development. Lectures given by leading SPM experts will be supported by supervised exercise classes in which experience will be gained in the solution of typical problems in SPM.
The Course will cover the basics of operation and advanced operation. Course registrants will have access to two STM and SFM microscopes in the Surface and Interface Reaction Group and much of the subject matter will be demonstrated on these instruments.
WHO SHOULD ATTEND
The course will be of maximum benefit to you if you are, or expect to be involved in using any form of Scanning Probe Microscopy as a research, diagnostic or trouble-shooting tool. Engineers, Chemists and Physicists who are using Scanning Electron Microscopy and Transmission Electron Microscopy will also find the course extremely useful.
COURSE FORMAT
The course will commence with registration at 9:30 on Monday 18 January 1999 and continue until 15:00 on Friday 22 January. The program of lectures is well distributed with a variety of tutorials each day. There will be plenty of opportunities for discussion with lecturers and other delegates. Full course notes will be supplied to all participants.
ORGANISERS AND PRINCIPAL LECTURERS:
Professor Jim Castle (SMME, University of Surrey) Dr. Peter Zhdan (SMME, University of Surrey)
INVITED LECTURERS INCLUDE:
Professor Michael Bowker (University of Reading) Professor Martyn Davies (University of Nottingham) Professor Trevor Page (University of Newcastle) Professor John Pethica (University of Oxford) Professor Richard Palmer (University of Birmingham)
For further information contact Dr. Peter Zhdan (P.Zhdan-at-surrey.ac.uk) or the Course Secretary:
Mrs. Margaret Morgan School of Mechanical and Materials Engineering University of Surrey Guildford, Surrey GU2 5XH UK
} -----Original Message----- } From: Steve Chapman [mailto:PROTRAIN-at-CompuServe.COM] } Sent: Thursday, September 17, 1998 12:04 AM } To: Melvyn Dickson } Cc: [unknown] Mel had written ... } } } } His answer was that users were opening the inner door of the airlock too } } soon and perhaps too quickly. There was too much air left in the airlock } } and the pressure rise in the chamber stalled the diffusion pump and } } backstreaming of the diffusion pump oil caused the contamination. }
Steve responded ... } } Do you believe this guy? } } All the work we did in the past proved to us beyond doubt } that the oil was from the rotary pump. No matter how } sophisticated the system is if you use } a rotary pump at all in the cycle you do seem to get RP oil as a } contaminant. } } I think the guy had a good try but I for one do not believe him! } } ...
I believe what the JEOL rep suggested was not out of line ... the contamination would still be from the RP if there were a slight DP burp.
What many JEOL users are not aware of is that (1) the "ready" lite responds to a timer not the interlock vacuum and (2) since JEOL started relying on a single RP (i.e., an RP is no longer 100% dedicated to the DP), the interlock pressure will rise after "ready" is indicated. I have to make sure all my users know this and that they should exchange the specimen immediately. Also (3) the interlock seal is subject to leaks because of dust, therefore the "ready" lite should be considered an indication of 90seconds only ... it is not an indicator that the interlock vacuum is ready. Given these possible problems, I wish JEOL would cap the DP for specimen exchanges ...
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
What are the lateral and Z dimensions of these structures? There have been a number of other techniques,including SWLI (Scanning White Light Interferometry) used to image these structures. I have access to a system which can do that, if it would be helpful.
I didn't think there was this much gold in the world to evaporate!
****************************** Jim Haley Applications Engineer I-CUBE 2411 Crofton Lane, Suite 14A Crofton, MD 21114 voice: (301) 858-0505 fax: (301) 858-0615 web site: http://www.i-cubeinc.com e-mail: haley-at-i-cubeinc.com ******************************
Timothy Moeller wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, Paul. } } Well, you've probably heard enough ideas on this subject. But I'll give } you my $0.02 worth (that's Au standard :-).) } } Working as a lab technician at UW Astrophysics Lab in Madison, WI, I } operated a vacuum deposition apparatus invented by a grad student which } vaporized gold wire and deposited it on the surface of insulated copper } wire to form the anode of a proportional detector. Anyway, in this } apparatus the gold wire was simply wrapped around tungsten (heater } filament) wire by hand. In vacuum, the tungsten filament was heated and } the gold wire melted, resulting in gold vapor being deposited evenly on } the wire rotissing nearby in the apparatus. It all seemed pretty crude } to me, but worked just fine. And I didn't take any special precautions } in wrapping the gold wire around the tungsten heater wire. } } Regs, } -- Tim } --- } } Gerroir, Paul J wrote: } } Hello Interested Readers, } } } } I have some experience in evaporating silver, (2 - 3mm shot) from a } } tungsten wire basket, however, I am now faced with challenge of evaporating } } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } } wire around the larger diameter tungsten wire and proceed or is there a } } better approach. Your comments/suggestions are appreciated. Thanks. } } } } Regards, } } Paul Gerroir } } Xerox Research Center of Canada } } -- } ...we now return control of your computer screen to you... } ------------------------------------------------------------ } Timothy G. Moeller | Microanalysis Products } Senior Software Engineer | NORAN Instruments, Inc., } {tmoeller-at-noran.com} | a ThermoSpectra company } ------------------------------------------------------------ } "I've spent my whole life trying to think up crazy ways of } doing things." } - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") } ------------------------------------------------------------
I am working with pollen and anther development of Ilex paraguariensis St. Hil. (Aquifoliaceae). However, the techniques used by me does not result in good ultrastrutural preservation. I am using Glutaraldehyde 2% + paraformaldehyde 2% in phosphate buffer ph 7.2, 0.1 M, at 4 oC, as a primary fixative. How obtain a "live-like" strutures in TEM? Which fixative should be used (concentrations, pH, osmolarity...)?
Thanks in advance M.Sc. Rinaldo Pires dos Santos Dept. of Botany - Universidade Federal do Rio Grande do Sul - UFRGS Av. Paulo Gama, 40 - Bairro Bom Fim - 90046-900 Porto Alegre - RS - Brazil e-mail: rinaldop-at-botanica.ufrgs.br
Dear Mel, This is a problem of all SEMs, not just JEOL, and really is a result of the EDS detector being the coolest spot in the chamber. The cleanliness of the vacuum system just regulates how long the contamination will take to build up. Link solved this problem by warming the snout of their detector. This doesn't solve the problem of oil contamination, it just moves it away from the EDS detector. I agree woth Steve Chapman that the oil is from the rotary pump. You wrote:
} Numbers of JEOL owners in Oz have complained of similar oil contamination. } When we last evaluated FESEMs the JEOL rep was persistent to know why we } didn't like the JEOL models. One of my reasons was the oil contamination } problem and I asked for some explanation as to why it was so common. } } His answer was that users were opening the inner door of the airlock too } soon and perhaps too quickly. There was too much air left in the airlock } and the pressure rise in the chamber stalled the diffusion pump and } backstreaming of the diffusion pump oil caused the contamination. } } } ***************************************************** } Mel Dickson, Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Beth, I have prepared CDs for SEM imaging and you must dissolve the plastic to get at the very thin aluminum foil that has the pits on it. The laser reads the pits through the plastic. I used tri-chlorethylene overnight to dissolve the plastic from a 1 square cm piece of the outside edge of a CD. In the morning I fished out this tiny scrap of Al foil and put it on an SEM stub. You can image either side and at 10,000 times see the little pits in their rows. These are pre-recorded CDs, the CD-Rs are quite different, since they use a laser-sensitive dye. You wrote: } } Hi, } I have a request for a micrograph of the pits and lands on a music CD. I } thought this would be fun and simple but we aren't having any luck imaging } anything (actually it was so reflective we imaged the detector!:-) } Here's what we've done: } We cut up a CD - using a piece of it close to the center hole (so we knew } it had something on it to see). The piece was mounted (lower surface up) on } a stub using a carbon sticky tab and silver paste. The sample was sputter } coated for 2 minutes (some for 3) and scoped. The first time we viewed it } we didn't coat it (see paragraph one for details). } } Do we need a solvent to munch awhile on the surface layer of the CD? } Any help would be greatly appreciated! } } It was a Christmas music CD so maybe that is the problem :-). } } beth Good luck, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
At the risk of further crowding your in-basket(s), I want to apologize to everyone for my recent mishap in posting an article (Re: Evaporation of Gold) which resulted in bouncing ad infinitum (or ad nauseum, if you prefer.) I believe the problem stems to a mistake I made in replying to the list on that subject. Apparently, you are not supposed to use the "Reply" function, but rather post a NEW message to the List Server at {Microscopy-at-MSA.Microscopy.Com} . So consider this a warning in addition to an apology -- it could happen to you as easily as it has happened to me. In fact, I'd seen this same thing happen to other posters as well recently, who apparently made the same mistake I did.
---------------------------------------------------------- Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ----------------------------------------------------------
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Hi, Luc et al.
Yes, having TWO (or more) tungsten filaments twisted together is important, to give the capillary action for more even dispersal as you mention. I forgot that important detail (it's been 27 years!) Thanks for jogging my memory. In fact, as I remember now, we were actually taking a long strand of tungsten wire and doubling it over more than once, to give a twisted set of 4 or more for our apparatus. We then wrapped the gold wire around that.
Regs, -- Tim
Luc Harmsen wrote: } Hi all. } Great topic. } We have been involved in building a new control unit for a factory which evaporates aluminium onto plastic parts to give them that silver look. } This has given us a lot more understanding of the coating technique as they have very large vacuum chambers and many samples that need an even and smooth coat each time. } What they do is to use tungsten wires coils that have two strands of W wire twisted together as aposed to, what we all seem to use, the single strand of W wire. The aluminium wire is then twisted by hand onto this coil, fairly loosely but just that it makes contact. The reasoning is that as you start the heating of the aluminium it will melt and flow, via capillary action, between the two W wires. This means that the aluminium is then in very good contact with the W coil and a lower current is needed to vaporise the aluminium. You also have a source of aluminium the whole length of the W coil each time. This ensures a even and repeatable coat each time. } } Cheers } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za } } -----Original Message----- } } From: Timothy Moeller [SMTP:tmoeller-at-noran.com] } Sent: Wednesday, September 16, 1998 7:15 PM } To: Paul.Gerroir-at-crt.xerox.com } Cc: Microscopy-at-sparc5.microscopy.com; Microscopy-at-sparc5.microscopy.com } Subject: Re: Evaporation of Gold } } Hi, Paul. } } Well, you've probably heard enough ideas on this subject. But I'll give } you my $0.02 worth (that's Au standard :-).) } } Working as a lab technician at UW Astrophysics Lab in Madison, WI, I } operated a vacuum deposition apparatus invented by a grad student which } vaporized gold wire and deposited it on the surface of insulated copper } wire to form the anode of a proportional detector. Anyway, in this } apparatus the gold wire was simply wrapped around tungsten (heater } filament) wire by hand. In vacuum, the tungsten filament was heated and } the gold wire melted, resulting in gold vapor being deposited evenly on } the wire rotissing nearby in the apparatus. It all seemed pretty crude } to me, but worked just fine. And I didn't take any special precautions } in wrapping the gold wire around the tungsten heater wire. } } Regs, } -- Tim } --- } } Gerroir, Paul J wrote: } } Hello Interested Readers, } } } } I have some experience in evaporating silver, (2 - 3mm shot) from a } } tungsten wire basket, however, I am now faced with challenge of evaporating } } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } } wire around the larger diameter tungsten wire and proceed or is there a } } better approach. Your comments/suggestions are appreciated. Thanks. } } } } Regards, } } Paul Gerroir } } Xerox Research Center of Canada }
-- .we now return control of your computer screen to you... ---------------------------------------------------------- Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ---------------------------------------------------------- "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ----------------------------------------------------------
Arthropods generally have problems like this, but they're easily solved by dissection. The problem is the cuticle.
You need to create as good a path for the fluids to flow through the critters as possible: pull off a leg and snip off the tarsus if you want to look at leg structures, etc.
Remove the opisthosoma and a couple of legs if you're interested in the prosoma, etc.
Phil
} Greetings, } } I would like to fix small spiders for TEM. I am not particular interested } in preservation of specific structures, but would like to preserve both } integumental and inner structures in general. I noticed that my specimens } floated, and I am afraid that the fixation will not penetrate properly. } } Does anybody has experience with fixation of similar specimens ? I would } appreciate your comments very much. } } Thank you. } } Regards } } Peter Funch } } ______________________________________________________________________ } Peter Funch } Assistant Professor, Ph.D. } } Department of Zoology Direct Line + 45 8942 2764 } Institute of Biological Sciences Secretary + 45 8942 2727 } University of Aarhus Telefax + 45 86 12 51 75 } Universitetsparken E-mail: } peter.funch-at-biology.aau.dk } Building 135 } DK-8000 Aarhus C } Denmark } ______________________________________________________________________
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net or poshel-at-hotmail.com
Does anyone know why some Polaroid shots from an SEM have faint parallel vertical lines with a consistant spacing of somewhat less than 0.5 mm? Are they in the film? I'm also getting patches of horizontal lines in some micrographs lately that I'm pretty sure aren't due to either charging or the Polaroid rollers. Could this be interference from the new lab next door?
Thanks,
Dee
____________________________________________________________________________ Note: Sometimes I don't receive incoming emails (with no notification to the sender). If I don't respond to your message, please send it again! ____________________________________________________________________________ _ Dee Breger Manager, SEM/EDX Facility Lamont-Doherty Earth Observatory Route 9W Palisades NY 10964 USA
To anyone who cares, After 50 hrs in 1% alconox the appearance and mechanical proper- ties of the Al foil seem to be unchanged. I didn't weigh the foil or look at it under a microscope, but anyone with too much free time is welcome to try it. Yours, Bill Tivol
I have some images of a CD master taken by SEM and AFM if you would like them. Drop me a line and I can send them as an attachement.
JB
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
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I am presently researching digital cameras which can be mounted on light microscopes for use for both fluorescence and bright field image capture. This is to augment film recording, not replace it. Resolution is very important as is even light spread. I am also interested in the different image storage mechanisms utilized by these cameras.
Any suggestions or personal experiences with using digital cameras for this purpose would be appreciated.
I promise to post a summary of responses.
Thanks, Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
} Does anyone know why some Polaroid shots from an SEM have faint parallel } vertical lines with a consistant spacing of somewhat less than 0.5 mm? Are } they in the film? I'm also getting patches of horizontal lines in some } micrographs lately that I'm pretty sure aren't due to either charging or } the Polaroid rollers. Could this be interference from the new lab next } door?
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
We make up a saturated stock bottle which we draw from, and replenish with UA and water (8-10g UA/100 ml) from time to time. The insoluble material is described in the Merck Index as being due to insoluble basic salts. It describes Uranyl Acetate as being "freely soluble in water acidulated with acetic acid" For years, we have followed a modification of a procedure from Millonig's 1976 book Laboratory Manual of Biological Electron Microscopy (pg 53) and added a few drops of acetic acid per 100mls of stock saturated UA (stored in a brown bottle). This seems to push the ppt reaction the other way and give a clear solution. There seems to be little difference in staining as long as only a few drops of acetic acid are used. Changing the pH of the stain by much, is risky though as there are numerous papers and procedures which modify the effects of UA stain by doing so. We have raised the pH to the 4.5-5.5 (any higher and the UA will ppt) and gotten improved staining but with unacceptable amounts of ppt on the sections.
When compared with the other chemicals in the EM lab, UA would seem to be relatively safe when used carefully. Ingestion and inhalation (exposure to dust) are our major concern due to heavy metal toxicity as well as the radiation hazards. Making sure that surfaces are not contaminated, and cleaning any spillage immediately from bottles and tables before it dries are important steps. Wearing gloves, and hand washing after glove removal are also important safeguards.
The radiation exposure hazard under most operating conditions seems minimal. The least exposure possible is desirable (ALARA), when you don't need to handle it, don't be near it. Using Bill's number's, you would still be well under the limits for occupational exposure if you were in constant contact with .6 millirem/hr for a 2000 hr work year, (correct me, but my references place the limits at 1.25 rem/quarter, 5 rem/year whole body and 18.75 rem/quarter, 75 rem/year for extremities (Rayburn)) The other factor to keep in mind is that we are not talking about a whole body exposure, but just exposure to the hands. All in all, the amount of exposure while making up and staining grids seems miniscule.
As an aside, the pretty flowered dinnerware from the 50's, the vivid oranges and yellows are from uranium. If you have any, run a Geiger counter over them, you'll be surprised the number of counts. Also the mantles from gas and propane lanterns contain radioactive thorium. In the past health physicist have suggested using them(sealed in their bags) for check sources for counters.
Regardless, because of the toxicity, radiation hazard, as well as expenses to purchase(well over $1.00/gm) and dispose of UA, minimizing the amount needed to be discarded and wasted seems desirable. To the extents possible, use of minimal amounts, and if considerable staining is done, making stock saturated solutions which can be diluted to the desired concentration as needed, are good ways to conserve UA, minimize radiation exposure, and inhalation and ingestion hazards.
Now, if we are starting a poll for the chemicals in the EM Lab that make us the most anxious, my vote is for cacodylate.
Excellent. We are also in the process of researching digital cameras for the same purpose. Any words of experience would be greatly appreciated. Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
On 17 Sep 1998, Debby Sherman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am presently researching digital cameras which can be mounted on } light microscopes for use for both fluorescence and bright field image } capture. This is to augment film recording, not replace it. Resolution is very } important as is even light spread. I am also interested in the different } image storage mechanisms utilized by these cameras. } } Any suggestions or personal experiences with using digital cameras for } this purpose would be appreciated. } } I promise to post a summary of responses. } } Thanks, } Debby Sherman, Manager Phone: 765-494-6666 } Microscopy Center in Agriculture FAX: 765-494-5896 } Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu } Purdue University } 1057 Whistler Building } West Lafayette, IN 47907-1057 } }
Beth- here is the technique I developed at UW (Seattle) with the help of a few of the gradstudents, it worked for me, hope it works for you. -Mike
Protocol for preparation of CDs for SEM analysis (by Mike Rock)
This simple method utilizes the coefficient of thermal expansion for separation of materials of differing densities. CDs are made up of a metallic core (usually aluminum or gold) surrounded by a plastic layer on either side. Other methods include dissolving the plastic with various solvents, or by removing the metal layer by etching techniques. Both may work fine, I have tried neither. This protocol uses liquid nitrogen to cool the sample (CD) to a point where the materials separate, and has proved successful with both gold and aluminum CDs.
Using tongues immerse the CD in the liquid nitrogen, after 15-30 seconds the CD will sound as if it is cracking. After 30- 60 seconds remove the CD from the liquid nitrogen.
Place the frozen CD on a firm surface and strike it with a hammer (wear safety glasses), the CD will shatter. Alternatively you may wish to slap the frozen CD down against the bench top (results of the two techniques are similar), shearing between the plastic and metal interface. The metal will easily pull away from the surface of the plastic if still in contact.
Mount the metallic layer, which contains the information tracks ("pits" and "lands") on a aluminum stub using double stick "conductive" carbon tape or tabs. Sputter coating is usually not necessary. Examination with the SEM is fairly routine at this point (5-15 kV).
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Hi, Luc et al.
Yes, having TWO (or more) tungsten filaments twisted together is important, to give the capillary action for more even dispersal as you mention. I forgot that important detail (it's been 27 years!) Thanks for jogging my memory. In fact, as I remember now, we were actually taking a long strand of tungsten wire and doubling it over more than once, to give a twisted set of 4 or more for our apparatus. We then wrapped the gold wire around that.
Regs, -- Tim
Luc Harmsen wrote: } Hi all. } Great topic. } We have been involved in building a new control unit for a factory which evaporates aluminium onto plastic parts to give them that silver look. } This has given us a lot more understanding of the coating technique as they have very large vacuum chambers and many samples that need an even and smooth coat each time. } What they do is to use tungsten wires coils that have two strands of W wire twisted together as aposed to, what we all seem to use, the single strand of W wire. The aluminium wire is then twisted by hand onto this coil, fairly loosely but just that it makes contact. The reasoning is that as you start the heating of the aluminium it will melt and flow, via capillary action, between the two W wires. This means that the aluminium is then in very good contact with the W coil and a lower current is needed to vaporise the aluminium. You also have a source of aluminium the whole length of the W coil each time. This ensures a even and repeatable coat each time. } } Cheers } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za } } -----Original Message----- } } From: Timothy Moeller [SMTP:tmoeller-at-noran.com] } Sent: Wednesday, September 16, 1998 7:15 PM } To: Paul.Gerroir-at-crt.xerox.com } Cc: Microscopy-at-sparc5.microscopy.com; Microscopy-at-sparc5.microscopy.com } Subject: Re: Evaporation of Gold } } Hi, Paul. } } Well, you've probably heard enough ideas on this subject. But I'll give } you my $0.02 worth (that's Au standard :-).) } } Working as a lab technician at UW Astrophysics Lab in Madison, WI, I } operated a vacuum deposition apparatus invented by a grad student which } vaporized gold wire and deposited it on the surface of insulated copper } wire to form the anode of a proportional detector. Anyway, in this } apparatus the gold wire was simply wrapped around tungsten (heater } filament) wire by hand. In vacuum, the tungsten filament was heated and } the gold wire melted, resulting in gold vapor being deposited evenly on } the wire rotissing nearby in the apparatus. It all seemed pretty crude } to me, but worked just fine. And I didn't take any special precautions } in wrapping the gold wire around the tungsten heater wire. } } Regs, } -- Tim } --- } } Gerroir, Paul J wrote: } } Hello Interested Readers, } } } } I have some experience in evaporating silver, (2 - 3mm shot) from a } } tungsten wire basket, however, I am now faced with challenge of evaporating } } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } } wire around the larger diameter tungsten wire and proceed or is there a } } better approach. Your comments/suggestions are appreciated. Thanks. } } } } Regards, } } Paul Gerroir } } Xerox Research Center of Canada }
-- .we now return control of your computer screen to you... ---------------------------------------------------------- Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ---------------------------------------------------------- "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ----------------------------------------------------------
This bounced once, I shall try again... (Nestor ignore the (not spam) email if this makes it to the listserver ok) ----------------------------------------------------------------
Well....
The Nuclear Regulatory Commission limits of skin & extremity (hands, feet) is 50 Rem per year. ...Not something to "shoot" for, since the dose is also limited to "As Low As Reasonably Achievable".
The (damage) conversion coefficient from mR from this source (no alpha if not ingested) to mRem is ~1. 50 Rem = 50,000 mRem. At a dose rate of 0.6 mR/hr, one would have to hold the container for many years to receive a one year maximum dose (50,000 / 0.6 per hr = max hours exposure). At 5 mR/hr, it would be 50,000 / 5. At that one would have to hold the container for 10,000 hours before exceeding NRC dose limits. Exposure will also decrease as a function of the square of the distance from the source.
For medical tests to discover any changes in body chemistry, it would take about 50 Rem acute whole body exposure.
Less dose is always better, but in realistic terms the dose from the UA should not be of any concern. If this level is of concern, do not fly in airplanes, live at high elevations, avoid all medical radiation, avoid certain beaches, beware of granite buildings, run from radium dial watches, etc. :)
The real danger is if the UA enters the body where the alpha source is in direct contact with livings tissue. Radiological bio-assay (urine/fecal) would be required to detect this.
} Using Bill's number's, you would } still be well under the limits for occupational exposure if you were in } constant contact with .6 millirem/hr for a 2000 hr work year, (correct me, } but my references place the limits at 1.25 rem/quarter, 5 rem/year whole } body
These limits are for radiation workers. Because we get paid, we can be exposed to a greater risk. The limit for the general population is 0.5 rem/year whole body, and I think this limit also applies to women who are or may be pregnant. I do not know the status of graduate students; I'd be inclined to err on the side of caution--especially since it is fairly easy to keep exposure to UA low. Yours, Bill Tivol
Look, I'm sorry. But this wasn't my doing. True, it stems from an oversight on my part (by posting to the wrong address, as I explained in my apology article), but I did NOT compound the error myself by re-sending the message, nor repeating the CC's, nor anything. The listserver did all that. This whole thing just blew up in my face, after I posted one simple little article incorrectly. I'm sorry, but it's out of my control.
Nestor -- please fix this at the server!
Everyone else -- please try to understand, I'm sorry!
You can improve membrane fixation in a couple of ways. The best I use is pos-fixation in 1% Uranyl acetate in bidestilled water = for 1hour (do not use Cacodilate or Phosphate buffers since the uranyl = acetate precipitates). This is done after the Osmium tetroxide fixation, = so, be carefull to wash these buffers out of the material before = applying the Uranyl acetate.=20 You can use if you wish 0.1M acetate acetic acid buffer. The pH should = be lower than 6.7 (? must check this value) since Uranyl acetate = precipitates at higher pH.
You can also add Potassium ferricyanide 0.5% to the osmium tetroxide and = fix longer than usual (up to 5 hours).
You may also have problems with the embeding medium. Some embeddings may = produce bad membrane preservation. I use Molenhauer's EPON-ARALDITE = mixture with good results.
Hope this helps
Dr. A.P. Alves de Matos Pathology Department Curry Cabral Hospital Lisbon apmatos-at-ip.pt
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} You can improve membrane fixation in = a couple of=20 ways. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} The best I use is pos-fixation in 1% = Uranyl=20 acetate in bidestilled water for 1hour (do not use Cacodilate or = Phosphate=20 buffers since the uranyl acetate precipitates). This is done after the = Osmium=20 tetroxide fixation, so, be carefull to wash these buffers out of the = material=20 before applying the Uranyl acetate. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} You can use if you wish 0.1M acetate = acetic acid=20 buffer. The pH should be lower than 6.7 (? must check this value) since = Uranyl=20 acetate precipitates at higher pH. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} You can also add Potassium = ferricyanide 0.5% to=20 the osmium tetroxide and fix longer than usual (up to 5 = hours). {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} You may also have problems with the = embeding=20 medium. Some embeddings may produce bad membrane preservation. I use=20 Molenhauer's EPON-ARALDITE mixture with good results. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D2} Hope this helps {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Dr. A.P. Alves de Matos {/FONT} {/DIV} {DIV} {FONT size=3D2} Pathology Department {/FONT} {/DIV} {DIV} {FONT size=3D2} Curry Cabral Hospital {/FONT} {/DIV} {DIV} {FONT size=3D2} Lisbon {/FONT} {/DIV} {DIV} {FONT size=3D2} {A = href=3D"mailto:apmatos-at-ip.pt"} apmatos-at-ip.pt {/A} {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {/BODY} {/HTML}
Group We hope to present the result of our salary survey in around a month. The hold up was caused by the lack of interest - particularly from this listserver. It has been, however, substantial from the readers of our publication and the MSA Conference attendees. If you are interested in the survey results, and have not yet provided your data (in absolute confidence), you are invited to do so. The survey includes only microscopists in the U.S. and excludes manufacturers and suppliers. Reponses may be made by return email or by fax (608-836-1969). As follows, your data should be in 8 fields: FIELD 1: Last Education Degree None/AA/BS/MS/PHD/MD FIELD 2: Years experience after graduation FIELD 3: Gender M - Male F - Female FIELD 4:Yearly income FIELD 5: Are you currently a supervisor/manager? Y - Yes N - No FIELD 6: Location. If in question, pick the area you feel closest to your own income level. MW - Midwest NE - Northeast SE- Southeast S - South W - West excluding California CA - California FIELD 7: Primary interest in: B - Biological Science P - Physical Science E - Earth Science FIELD 8: Now working in: I - Industry E - Education H - Hospital/Medical G - Government (as employee with GS Scale) GS - Government sponsered research
Immersion oil. Students do not always clean the oil off of the objectives after use, and thus we have suffered oil infiltration into some objectives. Recently one of our faculty members was advised to use type B immersion oil as opposed to type A oil. This due to the type objectives we are using, A.O. 100x oil and Olympus 100x oil. My questions, is there a chemical make up difference in these oils, or is it just a high-low viscosity issue. TIA tom_osborn-at-csubak.edu Tom Osborn Staff California State University Bakersfield
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Hi, Luc et al.
Yes, having TWO (or more) tungsten filaments twisted together is important, to give the capillary action for more even dispersal as you mention. I forgot that important detail (it's been 27 years!) Thanks for jogging my memory. In fact, as I remember now, we were actually taking a long strand of tungsten wire and doubling it over more than once, to give a twisted set of 4 or more for our apparatus. We then wrapped the gold wire around that.
Regs, -- Tim
Luc Harmsen wrote: } Hi all. } Great topic. } We have been involved in building a new control unit for a factory which evaporates aluminium onto plastic parts to give them that silver look. } This has given us a lot more understanding of the coating technique as they have very large vacuum chambers and many samples that need an even and smooth coat each time. } What they do is to use tungsten wires coils that have two strands of W wire twisted together as aposed to, what we all seem to use, the single strand of W wire. The aluminium wire is then twisted by hand onto this coil, fairly loosely but just that it makes contact. The reasoning is that as you start the heating of the aluminium it will melt and flow, via capillary action, between the two W wires. This means that the aluminium is then in very good contact with the W coil and a lower current is needed to vaporise the aluminium. You also have a source of aluminium the whole length of the W coil each time. This ensures a even and repeatable coat each time. } } Cheers } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za } } -----Original Message----- } } From: Timothy Moeller [SMTP:tmoeller-at-noran.com] } Sent: Wednesday, September 16, 1998 7:15 PM } To: Paul.Gerroir-at-crt.xerox.com } Cc: Microscopy-at-sparc5.microscopy.com; Microscopy-at-sparc5.microscopy.com } Subject: Re: Evaporation of Gold } } Hi, Paul. } } Well, you've probably heard enough ideas on this subject. But I'll give } you my $0.02 worth (that's Au standard :-).) } } Working as a lab technician at UW Astrophysics Lab in Madison, WI, I } operated a vacuum deposition apparatus invented by a grad student which } vaporized gold wire and deposited it on the surface of insulated copper } wire to form the anode of a proportional detector. Anyway, in this } apparatus the gold wire was simply wrapped around tungsten (heater } filament) wire by hand. In vacuum, the tungsten filament was heated and } the gold wire melted, resulting in gold vapor being deposited evenly on } the wire rotissing nearby in the apparatus. It all seemed pretty crude } to me, but worked just fine. And I didn't take any special precautions } in wrapping the gold wire around the tungsten heater wire. } } Regs, } -- Tim } --- } } Gerroir, Paul J wrote: } } Hello Interested Readers, } } } } I have some experience in evaporating silver, (2 - 3mm shot) from a } } tungsten wire basket, however, I am now faced with challenge of evaporating } } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } } wire around the larger diameter tungsten wire and proceed or is there a } } better approach. Your comments/suggestions are appreciated. Thanks. } } } } Regards, } } Paul Gerroir } } Xerox Research Center of Canada }
-- .we now return control of your computer screen to you... ---------------------------------------------------------- Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ---------------------------------------------------------- "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ----------------------------------------------------------
You probably have to cut the animals in a drop of fixative or inject the = fixative into the animal's body. If the animal is too small, I would try to injure the surface to provide = entry points to the fixative. Perhaps removing the legs?. The floating can be controled by pushing the animals into the fixative = bottle with a cotton plug, filter paper or something similar.
Dr. A.P. Alves de Matos Pathology Department Curry Cabral Hospital Lisbon apmatos-at-ip.pt
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} You probably have to cut the animals = in a drop=20 of fixative or inject the fixative into the animal's body. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} If the animal is too small, I would = try to=20 injure the surface to provide entry points to the fixative. Perhaps = removing the=20 legs?. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} The floating can be controled by = pushing the=20 animals into the fixative bottle with a cotton plug, filter paper or = something=20 similar. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Dr. A.P. Alves de Matos {/FONT} {/DIV} {DIV} {FONT size=3D2} Pathology Department {/FONT} {/DIV} {DIV} {FONT size=3D2} Curry Cabral Hospital {/FONT} {/DIV} {DIV} {FONT size=3D2} Lisbon {/FONT} {/DIV} {DIV} {FONT size=3D2} {A=20 href=3D"mailto:apmatos-at-ip.pt"} apmatos-at-ip.pt {/A} {/FONT} {/DIV} {/BODY} {/HTML= }
Interesting - I have always hit the 'reply' button, intending to send something just to the individual doing the posting, and that is exactly what has happened, and there has been nothing at all on the ListServer (never mind 'bouncing ad infinitum'). I have just now hit the 'reply all' button on our MS Exchange system for this sending (with the difference that the 'Microscopy-at-Sparc5.Microscopy.Com' address is there along with yours). So let's see what transpires, but I have a feeling the 'bouncing' has something subtle to do with your local stuff. I'm sure our esteemed Sysop would have posted 'How To Avoid' guidelines by now if it was as simple a matter as you say. (Or----, Nestor?).
tom
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 623-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: Timothy Moeller[SMTP:tmoeller-at-noran.com] } Sent: September 17, 1998 12:37 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: an apology (and a warning!) } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } At the risk of further crowding your in-basket(s), I want to apologize } to everyone for my recent mishap in posting an article (Re: Evaporation } of Gold) which resulted in bouncing ad infinitum (or ad nauseum, if you } prefer.) I believe the problem stems to a mistake I made in replying to } the list on that subject. Apparently, you are not supposed to use the } "Reply" function, but rather post a NEW message to the List Server at } {Microscopy-at-MSA.Microscopy.Com} . So consider this a warning in addition } to an apology -- it could happen to you as easily as it has happened to } me. In fact, I'd seen this same thing happen to other posters as well } recently, who apparently made the same mistake I did. } } ---------------------------------------------------------- } Timothy G. Moeller | Microanalysis Products } Senior Software Engineer | NORAN Instruments Inc., } {tmoeller-at-noran.com} | a ThermoSpectra company } ---------------------------------------------------------- }
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Hi, Luc et al.
Yes, having TWO (or more) tungsten filaments twisted together is important, to give the capillary action for more even dispersal as you mention. I forgot that important detail (it's been 27 years!) Thanks for jogging my memory. In fact, as I remember now, we were actually taking a long strand of tungsten wire and doubling it over more than once, to give a twisted set of 4 or more for our apparatus. We then wrapped the gold wire around that.
Regs, -- Tim
Luc Harmsen wrote: } Hi all. } Great topic. } We have been involved in building a new control unit for a factory which evaporates aluminium onto plastic parts to give them that silver look. } This has given us a lot more understanding of the coating technique as they have very large vacuum chambers and many samples that need an even and smooth coat each time. } What they do is to use tungsten wires coils that have two strands of W wire twisted together as aposed to, what we all seem to use, the single strand of W wire. The aluminium wire is then twisted by hand onto this coil, fairly loosely but just that it makes contact. The reasoning is that as you start the heating of the aluminium it will melt and flow, via capillary action, between the two W wires. This means that the aluminium is then in very good contact with the W coil and a lower current is needed to vaporise the aluminium. You also have a source of aluminium the whole length of the W coil each time. This ensures a even and repeatable coat each time. } } Cheers } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za } } -----Original Message----- } } From: Timothy Moeller [SMTP:tmoeller-at-noran.com] } Sent: Wednesday, September 16, 1998 7:15 PM } To: Paul.Gerroir-at-crt.xerox.com } Cc: Microscopy-at-sparc5.microscopy.com; Microscopy-at-sparc5.microscopy.com } Subject: Re: Evaporation of Gold } } Hi, Paul. } } Well, you've probably heard enough ideas on this subject. But I'll give } you my $0.02 worth (that's Au standard :-).) } } Working as a lab technician at UW Astrophysics Lab in Madison, WI, I } operated a vacuum deposition apparatus invented by a grad student which } vaporized gold wire and deposited it on the surface of insulated copper } wire to form the anode of a proportional detector. Anyway, in this } apparatus the gold wire was simply wrapped around tungsten (heater } filament) wire by hand. In vacuum, the tungsten filament was heated and } the gold wire melted, resulting in gold vapor being deposited evenly on } the wire rotissing nearby in the apparatus. It all seemed pretty crude } to me, but worked just fine. And I didn't take any special precautions } in wrapping the gold wire around the tungsten heater wire. } } Regs, } -- Tim } --- } } Gerroir, Paul J wrote: } } Hello Interested Readers, } } } } I have some experience in evaporating silver, (2 - 3mm shot) from a } } tungsten wire basket, however, I am now faced with challenge of evaporating } } Au wire onto a similar substrate. Is it appropriate to simply wrap the Au } } wire around the larger diameter tungsten wire and proceed or is there a } } better approach. Your comments/suggestions are appreciated. Thanks. } } } } Regards, } } Paul Gerroir } } Xerox Research Center of Canada }
-- .we now return control of your computer screen to you... ---------------------------------------------------------- Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ---------------------------------------------------------- "I've spent my whole life trying to think up crazy ways of doing things." - Chief Engineer Montgomery "Scotty" Scott (TNG:"Relics") ----------------------------------------------------------
Hi: I am looking for a used Zeiss 10C or Zeiss 109 for a customer of mine. The scope will be used in the Los Angeles area. Please respond directly to me. Thank you. Peter Jordan, EMSI 909 694-1839
The pits and lands on a CD are { {inside} } the plastic of the CD, so you need to dissolve it with some sort of solvent (I believe I used methanol, but can't recall for sure). The correct organic solvent will make the plastic disappear entirely, not just craze it and turn it cloudy. I think there may be several types of plastic used as the base, because I used toluene a number of years ago the first time I tried this, and it didn't work on the CD I
used a few months ago. Anyway, use a couple of fresh changes until the foil is floating free, then mount it on a stub and take a look. You might need to mount both sides of a single piece so that you are sure you have the side with the pits. If you've gotten all the plastic off, you might get away without coating, but I coated mine anyway.
You can see an example of the pits and lands at http://www.mta.ca/~jehrman/cd.htm
BTW, the CD I used is a Microsoft(TM) Office demo CD, so any flaws are Bill Gate's fault, not mine!
Cheers,
Jim
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Hello, all! This is proving to be a bit more "thorny" than I had expected. Thanks to those who have responded so far. Apparently there isn't any easy reference on TEM populations. Any or all additional inputs will be appreciated, and I WILL summarize and share the results! Thanks!
The repeating messages are NOT associated directly with an individual poster, so please don't complain to them or post your messages to the server it's not their fault as far as I can tell.
So far the only thing I have been able to determine for sure is that:
1.)" ALL " the repeating messages are definitely being bounced by a computer called "listserv.okstate.edu"
2.) "MOST" of those messages have "CC" microscopy
I have put the listserv.okstate.edu computer on the rejection list, which means if for some reason your Email routes through "okstate.edu" your posting may get rejected.
Do not "CC" microscopy. You should send all messages directly to our main address of
Microscopy-at-MSA.Microscopy.Com
while people have CC'ed Microscopy for several years, something has obviously changed recently (although not here) which causes CC'ed messaged which pass through listserv.okstate.edu to bounce.
My guess is that there is something strange going on at listserv.okstate.edu and I'm investigating. I seem to recall some recent requests from subscribers from that domain saying that their Email system has changed. Perhaps that is the source of the problem. I am investigating.
For now you will just have to bear with the problem.
The pits and lands on a CD are { {inside} } the plastic of the CD, so you need to dissolve it with some sort of solvent (I believe I used methanol, but can't recall for sure). The correct organic solvent will make the plastic disappear entirely, not just craze it and turn it cloudy. I think there may be several types of plastic used as the base, because I used toluene a number of years ago the first time I tried this, and it didn't work on the CD I
used a few months ago. Anyway, use a couple of fresh changes until the foil is floating free, then mount it on a stub and take a look. You might need to mount both sides of a single piece so that you are sure you have the side with the pits. If you've gotten all the plastic off, you might get away without coating, but I coated mine anyway.
You can see an example of the pits and lands at http://www.mta.ca/~jehrman/cd.htm
BTW, the CD I used is a Microsoft(TM) Office demo CD, so any flaws are Bill Gate's fault, not mine!
Cheers,
Jim
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
The pits and lands on a CD are { {inside} } the plastic of the CD, so you need to dissolve it with some sort of solvent (I believe I used methanol, but can't recall for sure). The correct organic solvent will make the plastic disappear entirely, not just craze it and turn it cloudy. I think there may be several types of plastic used as the base, because I used toluene a number of years ago the first time I tried this, and it didn't work on the CD I
used a few months ago. Anyway, use a couple of fresh changes until the foil is floating free, then mount it on a stub and take a look. You might need to mount both sides of a single piece so that you are sure you have the side with the pits. If you've gotten all the plastic off, you might get away without coating, but I coated mine anyway.
You can see an example of the pits and lands at http://www.mta.ca/~jehrman/cd.htm
BTW, the CD I used is a Microsoft(TM) Office demo CD, so any flaws are Bill Gate's fault, not mine!
Cheers,
Jim
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
If you've already checked the rollers in the Polaroid film back, the barring is probably on the CRT screen, and the next step is to calculate the frequency of the interfering signal. Simply divide the number of cycles of the barring in one scan line by the line time. If this comes out to 60 Hz, you probably have a ground loop, or an external magnetic field problem. If the frequency is 120 Hz, the problem is almost certainly ripple in the output(s) of your SEM's power supply. This may be a component failure (probably a capacitor, rectifier, or pass transistor), or could come from low line voltage. I've had problems with that on some older Philips machines, their power supplies really depend on line voltage being up to 100% of spec.
On Thu, 17 Sep 1998 micro-at-ldeo.columbia.edu-at-sparc5.microscopy.com wrote:
} Date: Thu, 17 Sep 98 13:33:55 EDT } From: "micro-at-ldeo.columbia.edu"-at-sparc5.microscopy.com } To: microscopy-at-sparc5.microscopy.com } Subject: sem image problems } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello collegues, } } Does anyone know why some Polaroid shots from an SEM have faint parallel } vertical lines with a consistant spacing of somewhat less than 0.5 mm? Are } they in the film? I'm also getting patches of horizontal lines in some } micrographs lately that I'm pretty sure aren't due to either charging or } the Polaroid rollers. Could this be interference from the new lab next } door? } } Thanks, } } Dee } } } ____________________________________________________________________________ } Note: Sometimes I don't receive incoming emails (with no notification to } the sender). If I don't respond to your message, please send it again! } ____________________________________________________________________________ } _ } Dee Breger } Manager, SEM/EDX Facility } Lamont-Doherty Earth Observatory } Route 9W } Palisades NY 10964 USA } } T: 914/365-8640 } F: 914/365-8155 } I: www.ldeo.columbia.edu/micro } } }
Robert Wieland wieland-at-me.udel.edu Neither Yankee nor Dixie, east of the Mason-Dixon line (look it up). You can't go faster than light, you can't get colder than absolute zero, and you can't help somebody by not telling them the truth.
Dee Berger wrote: Does anyone know why some Polaroid shots from an SEM have faint parallel vertical lines with a consistant spacing of somewhat less than 0.5 mm? Are they in the film? I'm also getting patches of horizontal lines in some micrographs lately that I'm pretty sure aren't due to either charging or the Polaroid rollers. Could this be interference from the new lab next door?
Dee, in 1987 I observed parallel lines, about 0.3mm separation, in Polaroid Type 52 film. The lines ran at a slight angle to the horizontal. Because the only appeared in one lot number of film and not others I blamed the film. By this time, however, we did not have much of that lot left and chose to ignore it. I still have copies those photos. I have not observed them since, but we have not used Polaroid film in quite a few years. Try exposing the film on an optical microscope. If you see them again, send the film and some of the photos to Polaroid. If it is their film, I'm sure they would like to know. If you don't see the lines, they are probably being produced in the SEM.
Dennis B. Barr (dennbarr-at-eastman.com) Physical Chemistry Research Laboratory ECC Physical & Analytical Chemistry Research Division B-150B, R-132E, (423) 229-2188
} -----Original Message----- } From: "micro-at-ldeo.columbia.edu"-at-Sparc5.Microscopy.Com } [SMTP:"micro-at-ldeo.columbia.edu"-at-Sparc5.Microscopy.Com] } Sent: Thursday, September 17, 1998 1:34 PM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: sem image problems } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Hello collegues, } } Does anyone know why some Polaroid shots from an SEM have faint } parallel } vertical lines with a consistant spacing of somewhat less than 0.5 mm? } Are } they in the film? I'm also getting patches of horizontal lines in some } micrographs lately that I'm pretty sure aren't due to either charging } or } the Polaroid rollers. Could this be interference from the new lab next } door? } } Thanks, } } Dee } } } ______________________________________________________________________ } ______ } Note: Sometimes I don't receive incoming emails (with no notification } to } the sender). If I don't respond to your message, please send it again! } ______________________________________________________________________ } ______ } _ } Dee Breger } Manager, SEM/EDX Facility } Lamont-Doherty Earth Observatory } Route 9W } Palisades NY 10964 USA } } T: 914/365-8640 } F: 914/365-8155 } I: www.ldeo.columbia.edu/micro }
I want to thank everyone who write to me about imaging a CD. The method that worked best (quickly, easily and no solvents!) was suggested by Brian McIntyre - see below. Just use a sticky tab on a stub to pull off the Al layer, sputter coat and scope. The images are nice. I also put some pieces of the CD in a solvent and got the Al layer that way, too. I apologize for ignoring the thread on CD prep recently. I do biological work and never thought I would need that info. So thanks to all who were willing to go there again.
best regards, Beth
} the surface you want to view is just under the top (label) side. the } easiest way to get the bit structure is to scribe a .5 X .5 cm square area } with a razor blade and then pull off the top plastic layer with the } underlying aluminum layer (with the bit structure replicated from the } bottom plastic layer) with a sticky tab on a metal stub.......this } technique works OK if you just want to see some pits and lands, it doesn't } however give a great whole piece sample. but this "problem" is surely } outweighed by its simplicity!! } } } good luck! } b- } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627 } } 716-275-3058 } 716-244-4936(fax) } "Be well, do good work, and keep in touch"
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Dear Timothy, } } Look, I'm sorry. But this wasn't my doing. True, it stems from an } oversight on my part (by posting to the wrong address, as I explained in } my apology article), but I did NOT compound the error myself by } re-sending the message, nor repeating the CC's, nor anything. The } listserver did all that. This whole thing just blew up in my face, } after I posted one simple little article incorrectly. I'm sorry, but } it's out of my control. } To err is human. To foul it up completely requires a computer. :-) Yours, Bill Tivol
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have received a number of responses to my original message (enclosed below) desiring info on a digital camera for LM however most have been from vendors. I would really like information from users.
I want to amend my original message with an additional requirement for a digital camera. Since I want maximum versatility, I do not want a system that requires hook-up to a computer during capture. I want a camera that can be easily moved for use on different microscopes which may not be in the same room. This would mean I need an image storage system like a PCMCIA card which can be used to collect the images and then read at a computer at another location. Our computers are in a room across the lab from the LM rooms and I do not want to tie up a network capturing images and transfering from afar. We also certainly do not need additional computers, nor do I want to take up resources or space for a computer dedicated to this camera. We also are a MAC lab so can handle images in TIFF format best as this can easily be dealt with on either computer platform.
I imagine that these requirements will really limit our choices but am shooting for everything and will worry about compromising if necessary later.
Thanks for the responses, Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Original message:
I am presently researching digital cameras which can be mounted on light microscopes for use for both fluorescence and bright field image capture. This is to augment film recording, not replace it. Resolution is very important as is even light spread. I am also interested in the different image storage mechanisms utilized by these cameras.
Any suggestions or personal experiences with using digital cameras for this purpose would be appreciated.
by nottingham.ac.uk with esmtp (Exim 1.92 #1) for Microscopy-at-Sparc5.microscopy.com id 0zJwpz-0000wY-00; Fri, 18 Sep 1998 10:28:23 +0100 Received: from CCW0F/SpoolDir by hermes.nottingham.ac.uk (Mercury 1.43); 18 Sep 98 10:28:24 GMT0BST Received: from SpoolDir by CCW0F (Mercury 1.43); 18 Sep 98 10:28:07 GMT0BST
Dear Microscopists I am looking for a bell jar (150mm diameter, 127mm high) for an Emscope SC500 coating unit. If any one has a spare they don't need and willing to part with please get in touch with me directly. We are able to pay for the item and shipping costs, but within the UK would be preferable.
Thanks a lot Nikki Nikki Bock EM Technician Dept. Materials Engineering University of Nottingham Nottingham NG7 2RD (0115) 9513759/9513871 Email: emznjb-at-hermes.nottingham.ac.uk
} We have several old American Optical/Spenser stereo microscopes that need a
} round glass stage insert. The inserts are 4" (10 cm) in diameter. I know
} we can have these made locally by a glass shop but was wondering whether
} anyone knows of an "off the shelf" source.
}
} Gary Radice
} Department of Biology
} Richmond VA
Gary,
These should probably be available through Leica. Also, you might want to contact a local service agent. Sometimes they know of sources which are more economical. If you don't have one in your area, contact me directly.
I recently had the same problem with faint parallel lines on Polaroid film with my SEM. This problem was not film-related and was simply due to electromagnetic interference from a nearby CRT. I had recently placed an X-terminal (with 20" monitor) on a table about 12 - 18 inches from the photo CRT on the SEM. The remedy was simple: either turn off the CRT when taking photos or move it away from the SEM console (3 - 4 feet was sufficient).
It sounds as though you may be having a similar problem and some kind of EM interference seems likely. Before shooting a Polaroid picture, try turning off devices around your SEM that are likely culprits, particularly any computer monitors, or ask the folks in the new lab next door if they will do the same. In lieu of that, you may be having an electronics problem with the microscope, such as high voltage instability, and your service person can help with that.
Hope this helps.
Dave
Dave Joswiak Dept. of Astronomy University of Washington Seattle, WA 98195
Hello collegues,
Does anyone know why some Polaroid shots from an SEM have faint parallel vertical lines with a consistant spacing of somewhat less than 0.5 mm? Are they in the film? I'm also getting patches of horizontal lines in some micrographs lately that I'm pretty sure aren't due to either charging or the Polaroid rollers. Could this be interference from the new lab next door?
Debbie Here's a summary of the most useful replies I received in response to my = inquiry about digital cameras. Naturally, most of the responses were from distributors, although many = had very informative content. Just the same, I left off their names in = the interest of fairness and maximizing S/N of this mailing list. I'm = happy to send unedited responses to anyone offline. Eric
------------ I think that there are two cooled CCD systems that you might consider. 1.) The first would be the Princeton Instruments MicroMax, a 12 bit, 5 = MHZ camera with a Grade 0 interline CCD, with thermoelectric cooling to = -10 C, which is also very fast as well, giving a full frame (1300 X = 1030) readout in 0.28 sec. This camera (including PCI interface) sells = for $19,500. 2.) An alternative system would be the Photometrics series 300, a 16 = bit, back illuminated camera using Grade 1, site 502B CCD, with = thermoelectric cooling to -25 C. This camera is not as fast as the = MicroMax, but has twice the QE, giving a full frame readout (512 X 512) = in 1. 4 sec, which may not be fast enough for your purposes. This camera = (including PCI interface) sells for $20,995. 3.) A third option would be the Quantix camera, also from Photometrics. = This is a 12-bit, 5 MHZ camera, using a Grade 1 KAF1400 CCD, also with = thermoelectric cooling (and the option for liquid cooling to -35 C). = This camera is similar in speed to the MicroMax, giving a full frame = readout (1317 X 1035) in 0.3 sec. This camera (with PCI interface) sells = for $24,495. One of the main differences between the first and third options is the = CCD chip. The interline chip is traditionally more sensitive in the = 'blue-green' region of the spectra than the KAF 1400 chip which is more = sensitive in the 'red' region of the spectra. Therefore if you will be = needing speed and sensitivity in the red as well as blue green part of = the spectra then the Quantix would be favored otherwise the MicroMax = would be favored. ---------- Olympus America sells a complete line of digital cameras for fura = applications. Currently we recommend either a frame transfer camera or = an interline camera for fura. These cameras are typically 12 - 14 bit = cameras with very good sensitivity for fura. Frame rates are variable, = typically from 1fps - 50fps. =20 You do not have to use ICCD cameras for fura. Digital cameras offer = several advantages over ICCDs. They have a greater dynamic range, = variable exposure rates and greater signal to noise ratio. ----------=20 I just received and quickly tested an Apogee KX2 - a cooled camera with = the Kodak 1300 chip. I am doing fluorescence work - although not = biological - and decided on this camera because of the dynamic range / = spectral range (I am using DAPI and IR filter sets) and price. Of = course it doesn't have quite the QE of a backilluminated camera, but on = first test it appears more than sufficient. Knowledgeable folks at that = company. Their first line of business is with astronomers but their = microscopy offerings are worth a look. Dave Calvert Eastman Chemical Co. P.O. box 1972 Lincoln Street=20 Kingsport, TN 37664 voice: (423) 229-4943 fax: (423) 229-4558 calvert-at-eastman.com ---------- I'm not sure what a "back-illuminated non-intensified" camera is, but I = would highly recommend a cooled CCD camera for your applications, = probably something like the Spot or maybe the Spot Jr., from Diagnostic = Instruments ($5-8K). =20 It depends on your application, but a 1/2 second exposure would be = pushing it for just about any CCD camera if you want color. There are = other high-end, high-speed digital cameras (maybe something from Dage = MTI) that could do this in B&W, but they are in the range of $20-25K.=20 ------------ We too are looking at digital cameras - had a demo of the olympus one = yesterday which I was impressed with! It did AO fluorescence on problem = - still have to try with other low light sources. problem with the olympus - nothing longer than 0.5s - and we regularly = take stuff of 2-3 s... ---------- Give our web on the new DVC 1300 digital cameras a review. It is very = detailed. http://members.aol.com/dvcco We can offer 1300 x 1030 pixels with integration / no cooling to 3 sec = or so with 10 bit S/N for a cost effective $4995 !!!! We are plug and = play with any RS-422 frame grabber board and we offer the boards also. = We are the US manufacturer of the camera and have taken on most board = lines for your convenience. ------------ Right now, the state of the art in Fura detectors is a Sony designed = chip family of progressive scan, interline transfer, Hyper HAD chips = like the ICX061 we use in our ORCA series cameras. These chips are used = by us and Princeton Instruments in cameras that produce high signal to = noise ratio images in low light conditions and are especially sensitive = in the blue wavelengths. The cameras cost about $15,000 and you will need a frame grabber and = software in addition. The total comes to about $20,000 usually. If = this is of interest to you, let me know and we will send you more = details. ------------ If you have to limit your exposures to 1/2 second, the SPOT camera will = not work as it is a real "Light Hog", and most exposures even with = bright fluorescence take 3-4 seconds per color channel. A color image = would then require 12-16 seconds to capture. Please call me at the = number below if you'd like to get pricing on some of our intensified CCD = cameras. We may be able to give you better pricing than you have been = seeing.=20 ----------- I sell a digital camera from the company Pixera. They did develop a = new digital camera. The sensitivity of this camera is 0.3 lux. You = can make frames accumulated or averaged. Max exposure time 12.8 sec. The complete system would cost about 6k. including a reduction lens for = the microscope.
For some work that we are doing we would like to know a source for Transine or Ammonium Fluoride. Does anyone know of a source? Thanks Al Bingham Semoptics Ltd. A manufacturer of Custom components for Scanning Electron microscopes.
I once saw a related problem in a Hitachi H2460N SEM, in which the Polaroid film would show lines at the top and bottom, but the middle would be clear. The lines faded out and back in again as the middle of the image was approached and passed. If memory serves me, the Hitachi service folks puzzled over this one and finally fixed it by adjusting the relationship of the X and Y screen scan controls. (If anyone from Hitachi is reading this maybe you can clarify/correct this?)
This particular artifact also showed up on the screen, however, while using the slower scan speeds.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML--Biology New Mexico State University Las Cruces, NM 88003
We are trying to embed relatively large (4mm x 5mm x 3mm) tissue samples in JB-4 and cut serial 5 micron sections for purposes of 3D reconstruction studies. The tissue of interest is the lamina cribrosa (connective tissue in eye). We are having a devil of a time for a variety of reasons, mostly related to my lack of expertise with JB-4. (For a complex set of reasons we cannot use paraffin.)
Can anyone help with the following questions? 1. The hardness of the resulting blocks is highly variable (maybe depending on the age of the catalyst?) The product insert states that more catalyst is needed as it ages, but gives no guidelines as to how much. Any ideas? 2. Sometimes the tissue sample becomes nearly transparent after infiltration, other times it does not. We cannot deduce why this occurs. 3. Some degree of curling and deformation of sections is inevitable. Any tips to reduce/minimize this?
Many thanks in advance, Ross Ethier -- Prof. C. Ross Ethier Department of Mechanical and Industrial Engineering, University of Toronto Toronto, Ontario M5S 3G8 email: ethier-at-mie.utoronto.ca voice: (416) 978-6728 fax: (416) 978-7753 http://mie.utoronto.ca/staff/profiles/ethier.html
I used to work in an ECM lab, so we had plenty of bottles of collagen lying about. Now that I'm ordering it for a new lab, I'm not sure if my predecessors just used what was available, or what was optimal.
When subbing slides (making them sticky so sections stay on), does it matter what sort of collagen is used? Is one particular type or bloom preferable,or just the cheapest? If the cheapest one is the answer, does anyone use Knox unflavored gelatin?
Thanks in advance.
Charlie Ginsburg NCC Research Dept. Lombard IL _________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
Dear Makroskopists, Someone asked me: what is the difference between the Wild Makroskop M420 and the M450? I know that the M420 is the M410 with with a trinoc or photo tube but I can find no info on the M450. Thank you.
########################### John Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901 Phone: 618-453-3730 Fax: 618-453-2665 ###########################
I think the "Gold" bouncing Email is done. If your interested read on..
First the bouncing mail was NOT the fault of either the Microscopy Listserver or of Timothy Moeller {tmoeller-at-noran.com} . If any of you vented on Tim, you owe him an apology.
The whole problem was a computer system at OKSTATE.
What apparently happened is that the system administrator's at OK State decided to rename a Email node from okway.okstate.edu to osu-com.okstate.edu. They did this without notice of the users and without setting up forwarding of Email from old to new system ID's.
The computer okway.okstate.edu serviced the EM facility and the Microscopy Listserver had 3 subscribers there.
In their infinite wisdom the SysOp's somehow mis configured things and mail did not forward from okway.okstate.edu to osu-com.okstate.edu but rather created the loop. Basically, their POP3 Server, which kept trying to receive mail for the now defunct computer okway.okstate.edu kept appending Microscopy-at-... to the CC: list of the mail which could not be delivered to the 3 people at the EM facility. Then, rather than just dying like a broken Email pipe sh ould have done, their's server forwarded mail to the CC list as if it originated at okway. This sent the mail back to Microscopy via the CC, which inturn sent it back to okway etc, etc, etc,.... hence the loop.
I would like to find out if and where there is an agent for PZO, the Polish Optical Company (other than in Poland). I am not having any luck corresponding directly and wish to contact an agent.
I think possibly the M450 was of the same series as the popular M400 but included some sort of built in illuminator. I am not positive but can probably look it up some time this week. If you have not already obtained an answer I can probably let you know sometime this week. -----Original Message----- } From: John J. Bozzola {bozzola-at-siu.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
The only digital camera for LM that does not require to be permanently connected to a computer I have heard of, is Olympus DP10. It uses SmartMedia card, although a permanent computer connection is an option. We do not have the camera, but considering to purchase it.
Alex ______________________________ Alexander Titkov
Millennium Inorganic Chemicals PO Box 245 Bunbury WA 6231 Australia Ph: (08) 9780 8505 FAX: (08) 9780 8500 E-mail: atitkov-at-micl.com.au
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I have received a number of responses to my original message (enclosed below) desiring info on a digital camera for LM however most have been from vendors. I would really like information from users.
I want to amend my original message with an additional requirement for a digital camera. Since I want maximum versatility, I do not want a system that requires hook-up to a computer during capture. I want a camera that can be easily moved for use on different microscopes which may not be in the same room. This would mean I need an image storage system like a PCMCIA card which can be used to collect the images and then read at a computer at another location. Our computers are in a room across the lab from the LM rooms and I do not want to tie up a network capturing images and transfering from afar. We also certainly do not need additional computers, nor do I want to take up resources or space for a computer dedicated to this camera. We also are a MAC lab so can handle images in TIFF format best as this can easily be dealt with on either computer platform.
I imagine that these requirements will really limit our choices but am shooting for everything and will worry about compromising if necessary later.
Thanks for the responses, Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Original message:
I am presently researching digital cameras which can be mounted on light microscopes for use for both fluorescence and bright field image capture. This is to augment film recording, not replace it. Resolution is very important as is even light spread. I am also interested in the different image storage mechanisms utilized by these cameras.
Any suggestions or personal experiences with using digital cameras for this purpose would be appreciated.
I'd really have to see the micrographs, but the vertical variations sound like some sort of electronic problem, although the frequency seems abnormally high. Can you provide more information in regards to the total record time and the number of lines of resolution used? Possibly the filtering of the accelerating voltage (usually around 20KHz) or the PMT high voltage (around the same frequency). In either case, the frequency would produce a vertical banding that would not be strictly vertical but at least slightly diagonal.
The horizontal variations may be a problem with the stage electrical connection. The sample is grounded through a connection that must remain constant, but in older instruments may vary. Normally a copper or copper-berylium spring contact is used that rubs against the sample holder through sample rotation. In older instruments, that contact may be flakey due to buildup of contaminants or reduction of spring force. Find the connection and clean both the spring loaded contact and the rotational surface it contacts with and try to reform the spring to produce a stronger contact.
} Hello collegues, } } Does anyone know why some Polaroid shots from an SEM have faint } parallel vertical lines with a consistant spacing of somewhat less } than 0.5 mm? Are they in the film? I'm also getting patches of } horizontal lines in some micrographs lately that I'm pretty sure } aren't due to either charging or the Polaroid rollers. Could this be } interference from the new lab next door? } } Thanks, } } Dee } } } ____________________________________________________________________ } ________ Note: Sometimes I don't receive incoming emails (with no } notification to the sender). If I don't respond to your message, } please send it again! } ____________________________________________________________________ } ________ _ Dee Breger Manager, SEM/EDX Facility Lamont-Doherty Earth } Observatory Route 9W Palisades NY 10964 USA } } T: 914/365-8640 } F: 914/365-8155 } I: www.ldeo.columbia.edu/micro } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Ken, I think you're wrong here. Silicone based oils will crack into an electrically non-conductive form in an electron optics column. The resultant contaminant coating will cause sample and optics component charging which will affect the electron beam. In my work, I suggest and use only Santovac DP oils, not just for their cracking characteristics to an electrically conductive film but also for their tolerance of large influxes of air while heated.
The ETEC instruments switched over, when properly calibrated, to the diffusion pump at 70 microns. This is appropriate to the range of a diffusion pump. But many other instruments switch over earlier, sometimes as high as 150 microns. In those SEMs, the early switch-over results in a stalling of the diffusion pump and a large release into the chamber of diffusion pump oils. As a case in point, I offer the Hitachi S-570. The preferred cure is a delay in the diffusion pump switch over to well under 100 microns.
Even in a well calibrated SEM, the delays in switching over to the diffusion pump and moderate air leaks can lead to a stalled diffusion pump. While sudies have shown that the primary contribution to most contamination problems is from mechanical pump oils, my practical experience is that contamination is often from diffusion pump oils. This also naturally leads to the question of what is an appropriate oil to use for mechanical pumps. Another thread?
} steve-at-facstaff.wisc.edu wrote: } } Dear all } } } } I may be dragging the thread away from contamination, but I } } thought that most electron microscope users avoided silicon based } } oils in diff pumps etc } } } } because they are extremely difficult to remove from the interior } } of an electron microscope and any contamination would normally } } have electrical insulating properties (catastrophic in an e.m.). } } Perhaps I am wrong but I would welcome any comments. After all } } this is one of the reasons why Santovac oils and their relatives } } became so popular (despite their costs). } } } } If I am labouring under a mis-apprehension then I apologise. } } } } Malcolm Haswell } } University of Sunderland } } UK } } } } All of the silicon based DP fluids I am aware of (which may not be } } all that are on the market) will break down under an electron beam } } and deposit a layer similar to glass on the nearest cool surface } } in the column. I don't know of any EM manufacturers that would } } recommend their use because they are almost impossible to remove } } if they do get in the column. We don't even use silicon based } } fluids in our vacuum evaporator. } } } } K. A. Brackett, Ph.D. } } TN & Assoc./USEPA } } } } Steve Limbach } } Associate Researcher } } Bock Research Lab. } } 1525 Linden Dr. } } } } UW-Madison } } Madison, Wisc. 53706 } } } } TEL 608 263-2582 } } FAX 608 262-4570 } } EMAIL slimbach-at- facstaff.wisc.edu } } Malcom, } I've been watching this thread with considerable interest, also. } ETEC sold almost every one of its microscopes with Transene Vacoil } (later Vacoil-S) which I believe is redistilled Dow-Corning 705. I } have never had any problems with it in 21 years of servicing these } instruments. Even a burped DP (and I've dealt with my share) has } never been a problem beyond the fact that you have to clean } everything. Many ETECs were sent out with the water-cooled baffle } plumbed in just before the DP rather than before the power supplies. } Those instruments will condense MP oil on their EDS detectors until } they get replumbed. but silicone doesn't seem to present any } problems. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
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} Dee, I had a recurring problem of vertical lines on the SEM image and Polaroids that was caused by noise in the digital logic circuits. This SEM had these logic circuits installed but not used. They would occasionally generate the vertical lines and the fix was to clean the circuit board "fingers" with and eraser. The boards were those in the CRT imaging bin. Hope this helps. Dave Audette audette-at-osi.sylvania.com } }
PZO has a website, with a possibility to send them an email trough that chanel. I have had some "bounced messages" some time ago. Perhaps the problem is solved now.
http://www.PZO.com
You'll find a link to their website in english there.
I don't know about PZO dealers in your country, their agent in Germany (and probably Europe...) is:
Gerhard G=F6ke Bahnhofstr. 27 D-58095 Hagen Germany. Telefon 02331/31754 Telefax 02331/31754
Dee, I have found the old Polaroids which exhibited parallel lines similar to those you wrote about. THEY WERE OPTICAL MICROGRAPHS. So the lines couldn't have been due to electronic interference. They were spaced at ~0.3 micron and were found throughout the image. We tried several different Polaroid film holders and got the same results with all of them, so we concluded the problem was not due to the film rollers. When we switched lots of film the problem went away.
Has anyone else ever seen lines like this?
Dennis B. Barr (dennbarr-at-eastman.com) Physical Chemistry Research Laboratory Physical & Analytical Chemistry Research Division Eastman Chemical Company Kingsport, TN 37662-5150
Debby - you are looking for a digital camera to suit your particular needs. It's a good starting point, but it is equally important to see what is available. Manufacturers of these instruments balance carefully customer's needs with what is technically possible and desirable. Professional digital cameras generally and those for microscopy in particular are tethered for good reasons. The non-tethered cameras for micrography most likely are toys. The problem is not just image storage, but extensive pre-photography software functions that are not available in any conventional camera. Most importantly the computer's screen serves as a quite superior viewer in all of these systems and, somehow, that requires a computer close to the camera. I cannot foresee this changing in the medium term. You could have a "roving" digital camera for your microscopes, by using a laptop, perhaps with a "real" screen attached. I expect, however, that in the long term it would be more practical to move the microscopes and to utilise a centrally located computer between a couple of superior microscopes. For class use, several 1 or 2 mega-pixel cameras could give students full-on experience in the new skills required with digital photography. The other points made: Uneven illumination relates to lenses or condenser system used and has nothing to do with the digital camera. Only "toy cameras" do not have the option to produce files in TIFF. Our online catalogue has a lot of non-camera specific digital photography information, particularly on the /epix.htm page I was not one of the vendors who wrote to Debby offering a digital camera, but would like to make the point that most vendors have much experience and most are honest. Sure, they may present a bias viewpoint, but don't expect to get an unbiased view from users either. They made a decision to buy a particular brand and they have experience with a particular range of jobs using that particular brand, well or badly. You can only hope for a diversity of views and that you have good judgement to pick A suitable system. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
I have received a number of responses to my original message (enclosed below) desiring info on a digital camera for LM however most have been from vendors. I would really like information from users.
I want to amend my original message with an additional requirement for a digital camera. Since I want maximum versatility, I do not want a system that requires hook-up to a computer during capture. I want a camera that can be easily moved for use on different microscopes which may not be in the same room. This would mean I need an image storage system like a PCMCIA card which can be used to collect the images and then read at a computer at another location. Our computers are in a room across the lab from the LM rooms and I do not want to tie up a network capturing images and transfering from afar. We also certainly do not need additional computers, nor do I want to take up resources or space for a computer dedicated to this camera. We also are a MAC lab so can handle images in TIFF format best as this can easily be dealt with on either computer platform.
I imagine that these requirements will really limit our choices but am shooting for everything and will worry about compromising if necessary later.
Thanks for the responses, Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Original message:
I am presently researching digital cameras which can be mounted on light microscopes for use for both fluorescence and bright field image capture. This is to augment film recording, not replace it. Resolution is very important as is even light spread. I am also interested in the different image storage mechanisms utilized by these cameras.
Any suggestions or personal experiences with using digital cameras for this purpose would be appreciated.
Dee, On my SEM if I use the quick scan to produce an image I get lines. When I ues the slow scan the lines are gone. It has to do with the way the recording monitor displays the image. Have you tried the different photo rates on the scope?
-- Regards, Gregory Rudomen Technical Specialist University Microscopy Imaging Center State University of New York at Stony Brook 516-444-3126 Greg-at-umic.sunysb.edu ********************************************************* Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. **********************************************************
Well, I guess that system is not OK after all. yuk, yuk, yuk.
Thanks Nestor for taking care of it!
Paula :-)
} Colleagues... } } I think the "Gold" bouncing Email is done. If your interested read on.. } } First the bouncing mail was NOT the fault of either the Microscopy } Listserver or of Timothy Moeller {tmoeller-at-noran.com} . If any } of you vented on Tim, you owe him an apology. } } The whole problem was a computer system at OKSTATE. } } What apparently happened is that the system administrator's at } OK State decided to rename a Email node from okway.okstate.edu to } osu-com.okstate.edu. They did this without notice of the users } and without setting up forwarding of Email from old to new system } ID's. } } The computer okway.okstate.edu serviced the EM facility and the } Microscopy Listserver had 3 subscribers there. } } In their infinite wisdom the SysOp's somehow mis configured } things and mail did not forward from okway.okstate.edu to } osu-com.okstate.edu } but rather created the loop. Basically, their POP3 Server, which } kept trying to receive mail for the now defunct computer okway.okstate.edu } kept appending Microscopy-at-... to the CC: list of the mail which could not be } delivered to the 3 people at the EM facility. Then, rather than just } dying like a broken } Email pipe sh ould have done, their's server forwarded mail to the CC list } as if } it originated at okway. This sent the mail back to Microscopy via the CC, which } inturn sent it back to okway etc, etc, etc,.... hence the loop. } } Once again we have been done in by someone else. } } Cheers.... } } Nestor } Your Friendly Neighborhood SysOp }
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
It was a great pleasure hearing Nancy Lane, a Cambridge cell biologist, speaking of the wonders of the microscope (including the electron variey) on the Classic FM programme "Masters of Their Art" (Saturday 19th September). This is Britain's largest commercial radio station.
Her work in particular uses invertebrates to research systems which have parallels in ourselves, such as the cell membranes that constitute the blood-brain barrier. Invertebrates generally reach us, at best, on the table, and so the "cruelty and cuddle" factors do not as yet restrict their use in experiments. But she deplored the low profile invertebrate studies have in biology courses, likening it to the situation where people talk of "shellfish" in a restaurant without regard to their nearly inexhaustible variety.
Still, such ignorance would be easily dispelled by a trip to Hong Kong, where I for one have expanded my gastronomic experience by at least two PHYLA, the echinoderms (sea cucumber) and the coelenterates (fried jellyfish).
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
At 09:00 AM 9/21/98 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm looking back a few years here, but I seem to recall a problem that was once caused by the rubber rollers in the Polaroid film holder (the one for P/N 55), rather than the metal rollers which squeeze the chemical packet. These rubber rollers had finely spaced parallel ridges (don't know the width) that can harden with encrusted chemicals if the unit is not frequently cleaned.
This doesn't answer as to why switching film lots caused the problem to go away, unless perhaps one lot had the quality of being ultra-sensitive to pressure. Anyone who has ever dropped a Polaroid film on the floor and stepped on it has seen the tread pattern of their sneakers on the final image.
This is probably reaching way out for an explanation, but your message did ring a bell. For what it's worth.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML--Biology New Mexico State University Las Cruces, NM 88003
Hi Charlie, we have always used the following mixture to coat slides so that plant sections (from paraffin embedments) stick:
Haup'ts adhesive
one gm Knox gelatin dissolved in 100ml dH2O at 30C. After dissolution of the gelatin add 2 gm phenol and 15 ml glcerin. Filter.
The phenol is added to prevent fungus from colonizing the gelatin but, as phenol is not nice, I have lately dispensed with it. Instead, freeze the mixture in aliquoits in Eppendorf tubes that can be thawed as required. Coat slides by rubbing 1-2 drops of the mixture onto the slide and letting sit until dry. Do not do this with bare fingers if there is phenol in the mix or ever if cleanliness is a concern. Hope this helps, John
} } I used to work in an ECM lab, so we had plenty of bottles of collagen } lying about. Now that I'm ordering } it for a new lab, I'm not sure if my predecessors just used what was } available, or what was optimal. } } When subbing slides (making them sticky so sections stay on), does it } matter what sort of collagen is used? } Is one particular type or bloom preferable,or just the cheapest? If } the cheapest one is the answer, does anyone use Knox unflavored gelatin? } } Thanks in advance. } } } Charlie Ginsburg } NCC Research Dept. } Lombard IL
________________________ C. John Runions Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
I have not seen MgO now for eight years, but I used to see a lot of them, together with other refractories. We compared grain-size by light microscopy. The problem with polishing with diamond paste only is that you get a perfect flat surface, without clear indications of grain boundaries, at least if you are dealing with 96+ % MgO, not the less pure ones. This prefect flat surface is probably what you mean by "smearing", because you expected grain boundaries. A simple method to make part of the grain boundaries visible is to do a last polish NOT with diamond paste, but something softer, such as alumina or aluminosilicate, the softer the better (and around 1 micron or finer). This gives relief polishing, boundaries becoming visible by differences in orientation . However, not all grain boundaries will be well visible, some boundaries have to be "filled in" by the observer.
A better visiblity of grain boundaries is provided by a thermal etch, but even here there is not 100% visibility of the grain boundaries. Saw a 1-2 mm section with the polished surface from the MgO (remove embedding material) and put that for 1/2 - 1 hour in a furnace at about 700C. These conditions are what I remember after 8 years, so I may be off slightly. With best regards, Emond de Roever
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I have been asked to compare the grain size of 2 MgO aggregates. The samples were embedded and polished with diamond paste, but I am not happy with the result. The surface looks smeared. Does the sample need to be etched or do I need to evaluate my polishing technique???? Thanks for you help.
Nan Laudenslager Specialty Minerals, Inc. Easton, PA nhl-at-early.com
by is2.nyu.edu (8.8.8/8.8.7) with SMTP id PAA01370; Mon, 21 Sep 1998 15:45:41 -0400 (EDT)
On Mon, 21 Sep 1998, Robert H. Olley wrote:
} Her work in particular uses invertebrates to research systems which have } parallels in ourselves, such as the cell membranes that constitute the } blood-brain barrier. Invertebrates generally reach us, at best, on the } table, and so the "cruelty and cuddle" factors do not as yet restrict } their use in experiments. But she deplored the low profile invertebrate } studies have in biology courses, likening it to the situation where people } talk of "shellfish" in a restaurant without regard to their nearly } inexhaustible variety.
I don't know your experience but invertebrates provide the substrate for much of classic and present research in basic neurobiology. And the advantages they provide are greatly appreciated and extolled in the standard texts.
} Still, such ignorance would be easily dispelled by a trip to Hong Kong, } where I for one have expanded my gastronomic experience by at least two } PHYLA, the echinoderms (sea cucumber) and the coelenterates (fried } jellyfish).
Or to modern Chinese and Japanese restaurants in the States.
} } I used to work in an ECM lab, so we had plenty of bottles of collagen } } lying about. Now that I'm ordering } } it for a new lab, I'm not sure if my predecessors just used what was } } available, or what was optimal. } } } } When subbing slides (making them sticky so sections stay on), does it } } matter what sort of collagen is used? } } Is one particular type or bloom preferable,or just the cheapest? If } } the cheapest one is the answer, does anyone use Knox unflavored gelatin?
If you simply want to make your slides sticky for paraffin or plastic sections, here is a protocol that works a lot better than polylysine, chromgel, or anything else I have ever used. It is also cheap!
PREPARATION OF COATED SLIDES Protocol 05.01.02 - last updated 9/5/95
Reference: In situ hybridization to cellular mRNAs using radioactively labeled RNA probes. L. Angerer (1989) In: In situ hybridization: methods for detecting DNA and RNA sequences at cellular and subcellular resolution. Am. Soc. Cell Biol. Workshop Manual. pp 1-21. Based on the more complex technique of Gotlieb and Glaser (1976) BBRC 63:815-821.
HAZARDOUS CHEMICALS: Do not use this protocol if you are not aware of all the safety precautions required to work with these chemicals. Consult all material safety data sheets. Work in a fume hood with proper safety technique. Wear appropriate safety gear.
SUPPLIES: 3 Coplin jars; 1 stock bottle; forceps. They need to be clean and lint free. Wear gloves when making the solution and dipping the slides.
1. Make a 2% 3-aminopropyltriethoxysilane(Sigma) working solution. Put 98 mls of acetone in the Schott bottle (orange cap) that is dedicated and marked for this purpose. Add 2 ml of 3-aminopropyltriethoxysilane. Pipet up and down a few times to mix the solution.
2. Fill the first Coplin jar with enough stock solution to cover the slide. Put the lid on unused stock. As the 3-aminopropyltriethoxysilane evaporates, refill Coplin jar with fresh stock. Fill a second Coplin jar with approximately 50 mls of acetone. Fill a third Coplin jar with approximately 50 mls of dH2O.
3. Line the work space in the fume hood with paper towels and place about three or four test tube racks on top of them as rests to lean the slides against after dipping.
4. To begin dipping the slides, you will need a pair of forceps. First, place between five and seven slides in the first jar (the acetone/aminopropyltriethoxysilane working solution). Wait about one minute before removing them.
5. Using the forceps, transfer these slides to the second Coplin jar (acetone). While you are letting these slides sit in the acetone for a minute, you can begin putting new slides into the first jar. If one is preparing large batches of slides, replace the acetone rinse periodically.
6. Transfer the slides in the second jar to the third jar (dH2O). Again, you will want to let the slides sit in the jar for about a minute. While waiting for these slides, you can continue to add slides to the first and second jars. If one is preparing large batches of slides, replace the dH20 periodically.
7. Before removing the slides out of the dH2O, you will want to dip them up and down a few times to make sure they have been rinsed well. Using the forceps, lean the slides against the test tube racks in order for them to dry.
8. Repeat this process until you have coated sufficient slides. When the slides are dry (about 10-15 min.), store them in a labeled slide box. The slides should be clear. White spots are a sign of improper rinsing.
9. When you are finished dipping the slides, clean up the hood. Do not save the stock solution longer than a couple of hours. Place the stock solution from the stock bottle and first Coplin jar into the appropriate hazardous waste bottle. Rinse these two containers with the acetone from the second Coplin jar. Discard the acetone into the appropriate hazardous waste bottle. You may dispose of the dH2O in the sink. The three Coplin jars and stock jar should be placed neatly off to the side within the hood to await future use. Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax) --============_-1305711192==_ma============ Content-Type: text/enriched; charset="us-ascii"
} } I used to work in an ECM lab, so we had plenty of bottles of collagen
} } lying about. Now that I'm ordering
} } it for a new lab, I'm not sure if my predecessors just used what was
} } available, or what was optimal.
} }
} } When subbing slides (making them sticky so sections stay on), does it
} } matter what sort of collagen is used?
} } Is one particular type or bloom preferable,or just the cheapest? If
} } the cheapest one is the answer, does anyone use Knox unflavored gelatin?
If you simply want to make your slides sticky for paraffin or plastic sections, here is a protocol that works a lot better than polylysine, chromgel, or anything else I have ever used. It is also cheap!
{bold} {bigger} {bigger} PREPARATION OF COATED SLIDES
{/bigger} {/bigger} {/bold} Protocol 05.01.02 - last updated 9/5/95
Reference: In situ hybridization to cellular mRNAs using radioactively labeled RNA probes. L. Angerer (1989) In: In situ hybridization: methods for detecting DNA and RNA sequences at cellular and subcellular resolution. Am. Soc. Cell Biol. Workshop Manual. pp 1-21. Based on the more complex technique of Gotlieb and Glaser (1976) BBRC 63:815-821.
HAZARDOUS CHEMICALS: Do not use this protocol if you are not aware of all the safety precautions required to work with these chemicals. Consult all material safety data sheets. Work in a fume hood with proper safety technique. Wear appropriate safety gear.
SUPPLIES: 3 Coplin jars; 1 stock bottle; forceps. They need to be clean and lint free. Wear gloves when making the solution and dipping the slides.
1. Make a 2% 3-aminopropyltriethoxysilane(Sigma) working solution. Put 98 mls of acetone in the Schott bottle (orange cap) that is dedicated and marked for this purpose. Add 2 ml of 3-aminopropyltriethoxysilane. Pipet up and down a few times to mix the solution.
2. Fill the first Coplin jar with enough stock solution to cover the slide. Put the lid on unused stock. As the 3-aminopropyltriethoxysilane evaporates, refill Coplin jar with fresh stock. Fill a second Coplin jar with approximately 50 mls of acetone. Fill a third Coplin jar with approximately 50 mls of dH2O.
3. Line the work space in the fume hood with paper towels and place about three or four test tube racks on top of them as rests to lean the slides against after dipping.
4. To begin dipping the slides, you will need a pair of forceps. First, place between five and seven slides in the first jar (the acetone/aminopropyltriethoxysilane working solution). Wait about one minute before removing them.
5. Using the forceps, transfer these slides to the second Coplin jar (acetone). While you are letting these slides sit in the acetone for a minute, you can begin putting new slides into the first jar. If one is preparing large batches of slides, replace the acetone rinse periodically.
6. Transfer the slides in the second jar to the third jar (dH2O). Again, you will want to let the slides sit in the jar for about a minute. While waiting for these slides, you can continue to add slides to the first and second jars. If one is preparing large batches of slides, replace the dH20 periodically.
7. Before removing the slides out of the dH2O, you will want to dip them up and down a few times to make sure they have been rinsed well. Using the forceps, lean the slides against the test tube racks in order for them to dry.
8. Repeat this process until you have coated sufficient slides. When the slides are dry (about 10-15 min.), store them in a labeled slide box. The slides should be clear. White spots are a sign of improper rinsing.
9. When you are finished dipping the slides, clean up the hood. Do not save the stock solution longer than a couple of hours. Place the stock solution from the stock bottle and first Coplin jar into the appropriate hazardous waste bottle. Rinse these two containers with the acetone from the second Coplin jar. Discard the acetone into the appropriate hazardous waste bottle. You may dispose of the dH2O in the sink. The three Coplin jars and stock jar should be placed neatly off to the side within the hood to await future use.
i've been doing some analysis of particulate matter filtered from urban and rural air using the old PRC program that tracor-northern sold in the '80s (boy do i feel old!). i have a few definition files setup to classify the particles, but i'm wondering if anyone out there has some files that work well for them that they'd like to share; i'd like to try some "better" or at least different classification schemes to see what i get. (this work is being done as part of a class project in environmental engineering for freshman undergraduates....its a hoot!)
thx!
b-
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax) "Be well, do good work, and keep in touch"
Hi, Recently I was quite unable to email the listserver. Some users may have had the same experience, so it seems worth posting. My attempted postings were returned with various computer generated messages, most commonly: "554 MX list for sparc5.microscopy.com points back to ultra.ultra.net.au" or "local configuration error"
"Points back", to me meant that the listserver, for whatever reason refused the email. I contacted Nestor via postmaster-at-microscopy.com with "I am not a spam" in the subject line. The short of it is that our service provider had changed their software about 10 days earlier. Amazingly, only about four out of dozens of email addresses caused problems but the fault clearly lay with our service providers software. Happily our genial and patient (going by emails only!) SysOp pointed me in this right direction. Thank you Nestor. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
} In theory, I expect that when compared with a tungsten } filament, at least the solid, single post LaB6 cathode } design by Kimball, would make for less movement, especially } during the warming-up period.
Just for the sake of fairness among us vendors, let me point out that both Denka (the LaB6 manufacturer which we represent in the United States) and FEI make LaB6 cathodes in which the crystal is mounted on rigid molybdenum posts, and this design is also recommended for microanalysis by virtue of its mechanical stability.
{snip}
Jim Darley added: } The obvious way to increase stability in LaB6 is to } increase the size of the microflat at the tip of the LaB6 } cone. The normal size of the cone (Kimball's) is 15 or 20 } micrometer. A 40 micrometer microflat is also available } (unfortunately at greater cost) and this makes a LaB6 quite } suitable for quantitative microanalyses. Brightness is } reduced but for microanalyses that is no consideration.
Again, this is product-specific. Denka, for example, produces "microflats" of 20, 40, 60 and 100 microns. It is also important to take cone angle into consideration. Both 60 and 90 degree cone angles are available, and the 60 degree cone angle has proven very useful for applications, like electron beam lithography, which require maximum stability.
Best regards, Steven E. Slap
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
Jim Darley wrote} In theory, I expect that when compared with a tungsten } filament, at least the solid, single post LaB6 cathode } design by Kimball, would make for less movement, especially } during the warming-up period. } In practise, tungsten filaments are more stable because the } emitting area is much larger and so, minor misalignment due } to a drifting filament is less consequential. } The stability we are talking about is due to slow drift and } this is of no consequence to normal, including high } resolution imaging in TEM or SEM. Stability over several } minutes matters when performing quantitative microanalyses } with a probe or EDS in dedicated SEM/TEM. In quantitative } analyses a spectrum maybe acquired over two minutes and } that is related to standard spectra and all numbers must } lead to results that come within 1% of reality. } The obvious way to increase stability in LaB6 is to } increase the size of the microflat at the tip of the LaB6 } cone. The normal size of the cone (Kimball's) is 15 or 20 } micrometer. A 40 micrometer microflat is also available } (unfortunately at greater cost) and this makes a LaB6 quite } suitable for quantitative microanalyses. Brightness is } reduced but for microanalyses that is no consideration. } Reducing downtime by increasing "filament life" to over } 5000 hours at low and medium emission is the main benefit } of a Lab6 in a microprobe. } For non-analyses work, a standard flat Lab6 cathode is } quite stable for SEM and TEM requirements; extreme drift } would matter but stability worries in these applications } are more likely to relate to HV instabilities or a pulsing } beam due to column/aperture contaminations. The question } for Mark Darus is the EM's vacuum system: Is it good enough } for LaB6 operation. } Disclaimer: ProSciTech supplies Kimball cathodes and } filaments.
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
We use Fisherbrand Superfrost/Plus Microscope Slides from FisherScientific. Cat # 12-550-15. They are more expensive than plain glass ones, but you save the trouble of having to mix up the collagen or p-L-lysine, worry about storage, or deal with contamination by fungus, etc. Sections never fall off, yet if you want to do in situ embedding, you can still get the resin slab off. I highly recommend them.
No commercial interest; just a satisfied customer.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I'll add my two cents regarding the discussion of Uranyl acetate radiation. Two years ago our Radiation Safety Office in Beltsville, MD asked our EM group to be responsible for the storage, etc. of all uranium compounds located at our site. I had heard the same comments in the past about not "needing to worry about UAc radiation". "Hey, the old reagent bottles don't even have radiation warnings on them"! "There is only a slight amount of alpha radiation".
Anyway, while gathering all uranium compounds, our radiation safety officer checked the compounds using a dosimeter that recorded alpha, beta, and gamma radiation. Levels of radiation were found that were just below OSHA guidelines for maximum daily exposure when the powder was checked from several inches away from the dosimeter. Thus, we decided to store the compounds in an acrylic box behind a beta shield in an isolated location. Whenever the actual Uranyl acetate is weighed to prepare dilute solutions proper safety precautions are recommended, i.e. use a beta shield, wear gloves and dust mask, weigh in a low occupancy room, etc.
We repeated our dosimeter readings last month with two different alpha, beta, and gamma dosimeters taking readings several feet from the bottles, removing the two acrylic shields one by one, and then with the cover of the UAc exposed (always moving closer to the chemical source). I won't enter the millirem/rad discussion; suffice to say it's an interesting little experiment for your support staff especially with the dosimeters left on audio signal. The dilute solutions themselves don't generate much radiation above background levels but should be treated as heavy metal wastes (don't pour down sinks).
Thanks for now.
PS Vote for most dangerous EM chemical DAB (diaminobenzidene).
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124
} ---------- } From: William Tivol[SMTP:tivol-at-wadsworth.org] } Sent: Wednesday, September 16, 1998 10:48 AM } To: michowej-at-nus.edu.sg } Cc: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Uranyl Acetate radiation } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Josephine, } } } } A solution of 10g UA in 15mls H2O was measured with a Geiger counter. } 500 } } counts/sec was generated. } } A supplier had measured 100g UA :- } } 1 Alpha - {2 counts/sec, using a 540 scintillation meter with AP-2 Probe } } 2 Beta - } 500 counts/sec, using a 540 E1 probe coupled to a GM Meter (this } } determines beta events and some low energy gamma events) } } 3 Gamma dose Rate (energy field) - two measurements done: } } using Mini monitor tpye R with GM Probe - 0.6mR/hr (mainly gamma) } } and Ionisation chamber DMM 95/0500 - 5 mR/hr (Beta and Gamma energy } } field). } } 4 Specific Activity (U approx. 55%) = 1.04 x 10 { {...} } Bq { {...} } gm } } { {...} } . } } } I calculated approximately the expected activity from 30 g UA } (about 0.1 mole, or 6*10^22 atoms). T_1/2 is 4.4*10^9 y and there are } 3.1*10^7 s/y, so the decay rate is 5*10^-18 s^-1, and the activity is } 3*10^5 Bq. } The build-up of daughter products with shorter half-lives will } reach steady state at which point the activities of the daughters will be } the same as that of the parent. Pa 234 has a gamma transition, and there } are several betas in the chain. The longer-lived isotopes in the chain } have lives of 10^4 to 10^5 y, and these will not be at steady state (unless} } your UA is *very* old ;-) ). 0.6 mR/hr is a significant amount of exposure, } and, if one were to hold the jars for some minutes, a sizable fraction of } the allowed annual dose would be attained. } } } Can UA be used openly without protection in laboratory? } } } Small amounts can be used, but be sure to wash hands before eating. } One area of the lab should be used for UA. A quiet area with little traffic } is best. UA, while not nearly the most dangerous EM reagent, should still } be treated with respect. } Yours, } Bill Tivol }
i have a collegue that just got a hold of a relatively old ( about 20 yrs ) spectrophotometric detector from perkin elmer model LC-75, however the opreating manuals seem to be missing. I realize that this is not a chromatography listserve, however, i was wondering if someone had any information that could help him out. He's contacted the manufacturer and they only sell manuals. He's a little hesitant to start purchasing material for this unit until he is certain that it can be of use to him.
thanks in advance
Michael Mandanas Particulate Materials Center 218 MRL Bldg. Pennsylvania State University University PArk, PA 16802 mxm67-at-email.psu.edu
Untethered, no cables required, camera, with high resolution? Full color?
There are VERY few. The best ones seem to be collaborations by Kodak with Nikon and Canon ( http://www.kodak.com/global/en/professional/products/cameras/range.shtml and http://www.kodak.com/US/en/digital/genInfo/EOSDCS1DCS460.shtml ). The Kodak DCS 460 (NIKON N90 camera body) and the Kodak EOS-DCS 1 (Canon EOS-1N body) were designed (1) to match the resolution of 35mm ISO 80-100 film; (2) be usable by professional photographers as to get the benefits of digital photography with the functionality and feel of an SLR camera. The Recording CCD records at 2036 x 3060 x 36bit (i.e. 18MB tiff files) with a sensitivity of ISO 80 (which is a little low for fluorescent work but usable). Both cameras utilize regular Nikon or Canon lenes, as well as the respective camera mounts for microscopes. "The images you shoot are stored on small, removable storage cards. A 170 MB card holds up to twenty-six 18 MB images. When you fill up one card, simply load another and keep shooting.After downloading image files from a storage card to your computer, you can even shoot over old files. Record more than 200 images per battery charge or an unlimited number if you're using the AC battery charger/adapter. " (You can get PC or Mac adapters to plug directly into your computer system.) Or you can capture directly via SCSI cable.
Sounds like what we're all looking for in a general LM camera? The problem is price Last time I got a price quote they were ~$13k. There are others available from Kodak with lower resolutions as well as high senesitivity (i.e ISO 200 - 1600 ), and I seemed to recall having read that Kodak is working on a new camera with both the high 2k x 3k resoltuion and high light senesitivity.
Sorry, since I still can't afford one, I can't tell you how well they handle LM. (Oh, I don't work for any vendor, I'm not selling anything, nor do I own stock in Kodak - at least I don't think I do.... : ).
So you see Virginia there is a Santa Claus (He's just really expensive).
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
Botanical Microtechnique and Cytochemistry by G.P. Berlyn and J.P. Miksche. Iowa State U. Press. 1976. ISBN 9138-0220-2
Nice book! Note, this is my copy, I don't know if there is a more recent edition, but Books In Print (at your local library or bookstore) should have the current information (you may need to check Forthcoming Books In Print). I used this text among others when I learned microscopy.
Phil
} Can anyone give me some references that pertain specifically to botanical } microtechniques? } } Preferably in english... } } Thanks! } } Yvan Lindekens.
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net or poshel-at-hotmail.com
Subbing slides. Make huge quantities at once using nothing but gelatin and water. (Additives may interfere with the stability of LM stains which engage in redox shifts with alacrity). Dry the slide, store dustfree, and throw out the gelatin mixture. Do not keep gelatin mixture even overnight. No problem with bacteria ever this way. The slides last forever. (0.1% gelatin in water is good, more is better ifyou can tolerate it. The thicker the gelatin coat the more interferece with photomigrography, but the more secure the section on the slide). So long, Hildy
i've been doing some analysis of particulate matter filtered from urban and rural air using the old PRC program that tracor-northern sold in the '80s (boy do i feel old!). i have a few definition files setup to classify the particles, but i'm wondering if anyone out there has some files that work well for them that they'd like to share; i'd like to try some "better" or at least different classification schemes to see what i get. (this work is being done as part of a class project in environmental engineering for freshman undergraduates....its a hoot!)
thx!
b-
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax) "Be well, do good work, and keep in touch"
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We recently had an interesting problem with vertical even spaced lines. We added a digital aquisition system to our JEOL JSM-840 SEM. All worked fine until we tried to capture an image using the compositional backscattering detector. The evenly spaced vertical lines appeared across the entire image. They did not appear under TOPO-BSI or SEI imaging of the same area.
Further investigation showed that the lines were still there, although the spacing changed, even when filament heating and KV were turned off as long as the mode selector was in COMP. It turned out that the problem was a wire from the COMP-SEI detector to ground. This wire had been there since the purchase of the microscope and we never could see a problem when recording COMP-SEI images using film. It only showed it's ugly side when recording using digital acqusition. The wire was removed and all is well.....not sure why it was there in the first place since there was other grounding as well. I do have a wonderful example of electrical interference for my SEM class so all is not a loss.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
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Hi,
We are planning to use DAB and nickel sulfate methods to colocalize synaptic proteins in wet sections. Question: If two proteins occur in the same area, does one stain cover the other? Has anyone had this experience? Does anyone have a better idea using wet sections and a light microscope (rather than a confocal, etc)? I would so much appreciate any opinions. Bye, Hildy {hcrowley-at-du.edu}
Greetings fellow microscopists; I have recently inherited a Leitz Dialux 22EB microscope, with attached camera. While I have little trouble with basic operation of a light microscope, the camera and setup are unfamiliar to me... is there anyone out there with the same or similar model who still has the manual, and would be willing to send me a copy? I have tried contacting Leica, who seem to have either been Leitz in a past life, or absorbed them somehow, but have not received a response so far. Any help would be most appreciated.
thanks in advance shea
Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
Hello fellow microscopists; I have inherited a Leitz Dialux 22EB light microscope, with an attached camera that is unfamiliar to me. The camera body says Wild MPS12 (Heerbrugg, Switzerland). While I suspect that I could eventually figure out how most of the stuff on the camera works (having been forced to do it with other scopes in the past) it would save me all kinds of time if someone has a manual kicking around that they could copy the pertinent bits for me from.
thanks in advance shea Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
The books that I use are john e. sass Botanical Microtechnique Plant Microtechnique by Johansen I will look for extra johansen... thought I had an extra one here...
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many months ago a collegue of mine asked to the microscopist from this listserver about tips and receipes por treating sand grains covered with a biofilm builded up by bacteria.
The procedure we followed was designed according to the literature and some of the answers we got from some of you. At the same time we followed instructions from someone who had done something like that before here. Neither of both gave us good results. My question is what we did wrong?????
These are some details of the 2 procedures we followed:
Procedure I
1) Primary fixation with glutaraldehyde 2,5% in buffer phosphate pH=6,7, at refrigerator temperature, 3 hs.
2) Three washes 10 or 15 minutes each in buffer phosphate
3) Secondary fixation with OsO4 2% in H2O, in the same buffer, at the same temperature, 1 h in darkness.
4) Dehydration in ETOH 50%, 70% and 95% once, 10' each, and 100%, twice or three times for 15' each.
5) Drying by filtering under vacuum on a 45 microns sterile Micropore filter. (we don't have facilities for crytical point drying)
Procedure II
The same steps except for the 3), that means only one fixation step (as done by someone else here before)
What we got is something strange (at least to us, who don't have any experience on treating biological samples). The grains treated according to Procedure II look very clear, transparent in some cases, and only a few are dark or grey. We think that is normal since the fixation treatment is not the appropiate one. Those treated by Procedure I look from light grey to dark grey and even black, just like the grains collected from the bioreactor that treats wastewater from a dairy products manufacture.
Nevertheless, on the SEM the biofilm supossed to be added on the grain surface is gone on both samples!!!! Where has it gone? Why that difference in colour then?
Please, we need to hear comments about this because we have no idea what has happened.
One more detail, the glutaraldehyde 25% we use looks light yellow, its pH is 3,7. If that is the problem, too much polimerysation, why do we still get dark grains?
Thanks to all of you in advance.
Silvia Montoro Centro Regional de Investigacion y Desarrollo de Santa Fe Santa Fe Guemes 3450 3000 Santa Fe Argentina csedax-at-arcride.edu.ar fax +54 42 550944
The wire was removed and all is well.....not sure why it was there in the first place since there was other grounding as well. I do have a wonderful example of electrical interference for my SEM class so all is not a loss.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
This may be a silly question. What is the smallest object ( using brightfield illumination) that can be observed. For example, I calculated resolution of .4 micron for a 100x, .90 NA objective and wavelengh value of 540. This is the smallest separation between features that can be resolved. Is this also the smallest feature that can be observed given a single feature on a smooth background?
Michael Ingram Rodel, Inc Polishing Lab 451 Bellevue Rd Newark, DE 19713 (302) 366-0500, ext.: 2545
Seems to me that I have seen in one of the Popular Science type magazines that a company called Imagek owned by Irvine Sensors Corp. is going to sell something called an EFS-1. It is a drop in cartridge for your 35mm camera body that is battery powered and capable of capturing mega pixel images. It will come with an adapter to upload your images to a computer and I think they were planning to sell for under $1,000. The neat thing is that you could still use all those expensive camera optics that you still have around. As usual I have no financial interest in this product, just something I thought I'd interject into this thread. I just checked and they have a web site at http://www.imagek.com/index.shtml.
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Smallest object depends on contrast: much smaller (than .4 micron) hole in thin specimen with transmitted light illumination will be visible.
Vladimir Dusevich
On Wed, 23 Sep 1998, Ingram, Mike wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This may be a silly question. What is the smallest object ( using } brightfield illumination) that can be observed. For example, I } calculated resolution of .4 micron for a 100x, .90 NA objective and } wavelengh value of 540. This is the smallest separation between } features that can be resolved. Is this also the smallest feature that } can be observed given a single feature on a smooth background? } } Michael Ingram } Rodel, Inc } Polishing Lab } 451 Bellevue Rd } Newark, DE 19713 } (302) 366-0500, ext.: 2545 } }
This may be a silly question. What is the smallest object ( using brightfield illumination) that can be observed. For example, I calculated resolution of .4 micron for a 100x, .90 NA objective and wavelengh value of 540. This is the smallest separation between features that can be resolved. Is this also the smallest feature that can be observed given a single feature on a smooth background?
Michael Ingram Rodel, Inc Polishing Lab 451 Bellevue Rd Newark, DE 19713 (302) 366-0500, ext.: 2545
This is not a silly question, and I have seen a suprising number of people get confused by this. Here is a simple test: go outside at night and look at the stars. Now, individual stars are below the resolution limit of the unaided eye (ex. you won't be able to resolve a binary star system). However, obviously you can see (detect!) them. This is because resolution limits and detection limits are two different things. Think of a single small bright object on a dark background. As the object approaches a true point, the image approaches the point spread function (by definition). There is no reason to expect that as the object drops below some (resolution) limit, the light coming from the object will stop propagating to the detector. As long as it is there is enough light reaching the detector, you can still detect it. The image doesn't get smaller (since the PSF is the limit, and it will get dimmer, but it still can be detected). Resolution simply involves seeing how close _another_ object can get to the first one without their images overlapping by some amount (that amount depending on whether you use the Rayleigh or Sparrow criterea). The perception or detection of a bright object on a dark background is limited by the "brightness" of the object. Now, the perception of a dark object on a bright background is different situation. Consider a dark line on a bright background: what happens when the line narrows and the two line spread functions get close to start to overlap? The minimum will become shallower and shallower. Based on the eyes' ability to discern intensity variations, Pluta (reference below) gave formulas for the detection of these objects: Using typical parameters (488nm, 0.9NA) the smallest dark objects on a bright background that can be observed are 41nm for a disk, and 2.6nm for a line.
Resolution, perception/detection and location are different, but unfortunately all three tend to get lumped together as "resolution". I know you didn't ask, but since it's related, you can also locate an isolated small object to better than the resolution limit (if you know that your system has at worst only symmetric aberrations). Try this: draw a circle and then try to find the center. Remember, this is not a random processes, but a deterministic one. This also extends to edge location. One study (can't find the reference right now) back in 1986 showed that confocal microscopes (of that era) had a 20nm uncertainty in locating edges (the same value as for the SEMs of the day). This means that as long as a the two sides of a line object are resolved, then you can measure the width of the object to much better than the resolution limit. Obviously SEMs can perform the measurements on narrower objects than LM, but I did included the qualification concerning the object being wide enough for the edges to be resolved in the previous sentence. (No flames for saying that LM will replace EM: I stated the limitations twice! (and pointed it out again)) Again, resolution, perception/detection and location are different (related to the imaging system, but still different).
Regards, Matt Atkinson 3M Corporate Research Labs
(Pluta's book Advanced Light Microscopy, vol 1, pg337-348 gives a very good explanation.)
mingram-at-rodel.com wrote:
} This may be a silly question. What is the smallest object ( using } brightfield illumination) that can be observed. For example, I } calculated resolution of .4 micron for a 100x, .90 NA objective and } wavelengh value of 540. This is the smallest separation between } features that can be resolved. Is this also the smallest feature that } can be observed given a single feature on a smooth background? } } Michael Ingram } Rodel, Inc } Polishing Lab } 451 Bellevue Rd } Newark, DE 19713 } (302) 366-0500, ext.: 2545
Can anyone point me in the right direction concerning the preparation of inorganic colloid samples in moderate/high ionic strength waters for quantitative analysis by TEM? The dispersions contain relatively low number concentrations of particles which we are trying to measure.
So far, samples have been prepared by ultracentrifuging onto carbon films (based on published method of Nomizu et al. Electron microscopy of nanometer particles in freshwater, Anal. Chem. vol 60, 2653-2656, 1988) and wicking the residual solution away with a filter paper. The problem is that there are considerable drying artefacts such as matting of particles, uneven dispersion of particles within a grid square (possibly due to surface tension effects from residual water films during drying?) and crystallisation of residual salts.
I'd be most interested to hear from anyone with experience of preparation of this type of sample or any good ideas.
Thanks in anticipation,
Simon
Dr Simon Dumbill Team Leader, Materials Characterisation AEA Technology 220, Harwell Tel: +44 1235 434245 Didcot Fax: +44 1235 435941 Oxfordshire OX11 0RA Email: Simon.Dumbill-at-aeat.co.uk UK
May I humbly suggest the following recipe: 0.5% gelatin (300bloom, nothing fancy), 0.05% chrome alum. Clean slides with a good detergent, rinse extensively with tap, then d.i.H2O. Sprinkle gelatin on cold H2O, warm carefully until gelatin is dissolved (do not boil!), add chrome alum*. Do NOT add the chrome alum before the gelatin. You may dip the slides in warm or cool. Allow to air dry, loosely covered from dust. I do not store the solution.
I do use the fancy Fisher slides for in situs, but NOT for free floating sections. Free floaters are thicker than what is recommended for those slides and sometimes do not adhere well to them. *chrome alum=chromium potassium sulfate
Thank you Caroline for the message. Coincidentally, I have two stereo SEM books for children 8 years and up coming into the book stores in November. One is called "Bug Eyes and Butterfly Wings" and the other is called "Snail Tongues and Spider Fangs."
They come with stereo glasses. Big kids should enjoy them too! My co-authors are Shar Levine and Leslie Johnstone who have published quite a number of Science books for kids.
ISBN: 1-894042-16-6 and 1-894042-05-0 Publishers: Canada - Somerville House, USA - Andrews McMeel, New Zealand - David Bateman
If anyone is interested in SEM puzzles, check out "Small Wonders" on -at- Discovery Canada on the Discovery Channel every Monday night 8pm-9pm. Unfortunately, I don't think -at- Discovery Canada is shown anywhere but Canada, but you can still participate in the puzzle by going to the Discovery Channel web site at www.exn.ca. Then go to -at- Discovery and then "Small Wonders." It is an interesting web site for all sorts other science stories.
} } Are any of you interested in the possibility of using your micrographs to } illustrate children's books? Here's a conference that should be quite } informative: } } Marine Biological Laboratory Hosts Institute for } Children's Book Authors and Illustrators } October 9-11, 1998 } } Woods Hole, MA-The Marine Biological Laboratory's Science Writing } Fellowships Program and the Center for Children's Environmental } Literature is co-sponsoring an Author, Illustrator, Biologist } Institute. Working and aspiring children's book authors and } illustrators, as well as scientists, are invited to participate in the } three-day meeting. Organizers hope to foster new collaborations between } authors, illustrators, and biologists. } } } For more information, contact: } Pamela Clapp Hinkle } Director of Communications } Marine Biological Laboratory } 7 MBL Street } Woods Hole, MA 02543 } Tel: 508-289-7276 } Fax: 508-457-1924 } e-mail: pclapp-at-mbl.edu } } For a complete program, see http://www.mbl.edu/html/MISC/AIB.html. } } Caroline Schooley } Educational Outreach Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
Some years ago I heard an account of a radiation safety inspection as part of a larger safety review at a large lab. The lab was involved in coal research, as were we, and had a fair amount of coal samples stored in glass jars. An inspection team came thru and happened to check the jars for radiation, found some, and instructed the researchers that they would need to start following radiation safety procedures.
Now coal can contain minute amounts of uranium in its mineral matter, but not enough that should set off a detector. Besides the detectors were setup to meaure alpha particles, and there was no way that alpha particles emitted from the contents of a glass jar should be detected on the outside.
A little digging revealed that there was in fact a little radiation present, but it was a slight residue left on the jar surface after washing. Apparently the jars went through the same washer as did other jars which had held radioactive materials. I think the radiation safety folks may have received additional training after the incident.
} Anyway, while gathering all uranium compounds, our radiation safety officer checked the compounds using a dosimeter that recorded alpha, beta, and gamma radiation. Levels of radiation were found that were just below OSHA guidelines for maximum daily exposure when the powder was checked from several inches away from the dosimeter. Thus, we decided to store the compounds in an acrylic box behind a beta shield in an isolated location. Whenever the actual Uranyl acetate is weighed to prepare dilute solutions proper safety precautions are recommended, i.e. use a beta shield, wear gloves and dust mask, weigh in a low occupancy room, etc. } } We repeated our dosimeter readings last month with two different alpha, beta, and gamma dosimeters taking readings several feet from the bottles, removing the two acrylic shields one by one, and then with the cover of the UAc exposed (always moving closer to the chemical source). I won't enter the millirem/rad discussion; suffice to say it's an interesting little experiment for your support staff especially with the dosimeters left on audio signal.
{snip}
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
Osmium is a contrast agent hence your black grains. As to the disapperance of the biofilm, I suspect that it is still there (at least to some degree), but that you cannot recognize anything biological because everything has completely collapsed/debrided during the final dehydration (essentially air-drying made even worse by the filtration under vacuum). I doubt you will see anything meaningful unless you find a critical-point dryer, particularly if the biofilm is rather thick to begin with. Bacterial cells are rather hardy, but there are good reasons why critical-point drying and other fancy methods were developed.
On the other hand, just what exactly are you trying to show by doing the SEM? If it is simply the presence of a biofilm, then why not use epifluorescence microscopy using some vital stains? It is a lot simpler and will give you more meaningful images than will SEM.
Rob Palmer CEB/UT
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ingram, Mike wrote: } } This may be a silly question. What is the smallest object ( using } brightfield illumination) that can be observed. For example, I } calculated resolution of .4 micron for a 100x, .90 NA objective and } wavelengh value of 540. This is the smallest separation between } features that can be resolved. Is this also the smallest feature that } can be observed given a single feature on a smooth background? } } Michael Ingram } Rodel, Inc } Polishing Lab } 451 Bellevue Rd } Newark, DE 19713 } (302) 366-0500, ext.: 2545
-- Not a silly question; I have a similar question along these lines... Correct me if I'm wrong, but don't we usually just use the Rayleigh criterion as a rule of thumb for estimating resolution limit in any optical system such as this?
---------------------------------------------------------- Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ----------------------------------------------------------
} Can anyone point me in the right direction concerning the preparation } of inorganic colloid samples in moderate/high ionic strength waters for } quantitative analysis by TEM? The dispersions contain relatively low } number concentrations of particles which we are trying to measure. } } So far, samples have been prepared by ultracentrifuging onto carbon } films (based on published method of Nomizu et al. Electron } microscopy of nanometer particles in freshwater, Anal. Chem. vol } 60, 2653-2656, 1988) and wicking the residual solution away with a } filter paper. The problem is that there are considerable drying } artefacts such as matting of particles, uneven dispersion of } particles within a grid square (possibly due to surface tension } effects from residual water films during drying?) and } crystallisation of residual salts.
I work on similar things and have a number of methods.
Firstly, you can just proceed by dipping a copper grid with supported carbon film into the colloid (if fairly dilute in particles) or a diluted version of the colloid. The carbon film is then left to dry on some filter paper. This still gives problems of clumping and crystallisation of salts on drying but may be the only way in some cases.
Alternatively, one can filter the particles from the colloid (if they will filter and not just go straight through the filter paper), wash with water, and redisperse in an organic solvent such as methanol. Then dip a copper grid with a supported carbon film into the suspension. This avoids the salt problem and does give better particle dispersion on the grid although it is not perfect and some particle clumping always occurs. The concentration of the suspension is critical here: too high and lots of clumping is inevitable, too low and the concentration of particles on your carbon film is rather low. Only trial and error will give the best concentration for your taste.
Finally, one could centrifuge the particles out, pour off the aqueous solution, add methanol or ethanol and redisperse and proceed as above. This is an alternative for the case where the particles are impossible to filter off.
Hope this helps.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, or: ianmaclaren-at-hotmail.com Birmingham B15 2TT, http://web.bham.ac.uk/I.MacLaren England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
After your final 100 ETOH rinse, try putting the samples in HMDS (hexamethyldisilazane) for about 30 minutes then keep in a desiccator for another 24 hours before viewing.
kim
} } 4) Dehydration in ETOH 50%, 70% and 95% once, 10' each, and 100%, twice or } } three times for 15' each. } } } } 5) Drying by filtering under vacuum on a 45 microns sterile Micropore filter. } } (we don't have facilities for crytical point drying) } }
} } } } Silvia Montoro } } Centro Regional de Investigacion y Desarrollo de Santa Fe } } Santa Fe } } Guemes 3450 } } 3000 Santa Fe } } Argentina } } csedax-at-arcride.edu.ar } } fax +54 42 550944 } }
HILDEGARD CROWLEY wrote: } } Hi, } } We are planning to use DAB and nickel sulfate methods to colocalize } synaptic proteins in wet sections. Question: If two proteins occur in } the same area, does one stain cover the other? Has anyone had this } experience? Does anyone have a better idea using wet sections and a light microscope (rather than a confocal, etc)? I would so much appreciate any opinions. } Bye, } Hildy } {hcrowley-at-du.edu}
Hildy:
I hate to say it all depends but it all depends. I have had good luck with doing DAB+Ni-Co first then DAB 'plain' for the second Ab but I was staining two different cell populations. When I was doing 2 Ab on one cell my results were mixed. Poor results when doing 2 monoclonals in one cell, good results when doing one mono and one poly. In an area as small as a synapse you might want to use two fluorescent markers, this usually gives very good results and can be very beautiful.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Anybody have a good protocol for preparing coccolithophores for taking pix of the intact coccospheres? I have fumbled around and obtained a few pix, but the yield of coccospheres is miniscule; these organisms seem to want to shuck their liths if you look at them cross-eyed.
I have a polarizer which is delaminating from the base glass substrate. This is in an rotating assembly specifically built as an Analyser for an older optical microscope.
Does anyone know how to either repair this or know where replacements can be obtained? I am assuming that the polarizer material is a linear polarizer - is this a fair assumption?
FYI: The polarizing material appears to be a thin platic-type material which has been glued to an underlying glass substrate. This sandwich is then mounted between 2 outer glass plates which have been (AR?) coated (or are these 1/4 wave plates?).
Any great ideas on how to recover this? Much thanks in advance - Ian L.
Wow! Thanks to everyone who responded so speedily to my cry for help. Sorry about the dual message... I tried to recall the first one, as it was only a day or so since I had tried to reach Leica via their web site. Leica (in the person of Philip Hyam) was in fact quite speedy in contacting me and assuring me of the availability of the manual.
Hi everybody! For one of our projects we used what we called crustacean buffer: 500mM NaCl, 12 mM KCl, 12mM CaCl2, 20 Mm MgCl2, 10 mM Tris (no HCl), and 4.3 mM Maleic Acid. We made double strenght buffer and mixed it with 4% aqueous sol. of osmium to obtain 2% working solution We did the same with glutaraldehyde: double strenght buffer with 10% aqueous glut to get 5% glut. Osmium solution truned yellow to brown within 1 hour after mixing and glut change color to yellow overnight. Needless to say fixation of the tissue was not great. Is there an expanation for that fenomena? I found a reference of maleate buffer used in staining procedures but not for fixation. Thank you. Dorota
} This may be a silly question. What is the smallest object ( using
} brightfield illumination) that can be observed. For example, I
} calculated resolution of .4 micron for a 100x, .90 NA objective and
} wavelengh value of 540. This is the smallest separation between
} features that can be resolved. Is this also the smallest feature that
} can be observed given a single feature on a smooth background?
}
} Michael Ingram
} Rodel, Inc
} Polishing Lab
} 451 Bellevue Rd
} Newark, DE 19713
} (302) 366-0500, ext.: 2545
Dear Michael,
The typical formula for resolution,R = (1.22 x lambda)/(NA objective + NA condenser), takes into account the following information:
(1) the nature of the background illumination, (2) orientation of the feature, (3) spacing (in your case, edge to edge) and (4) edge fidelity. Your calculation is correct for the smallest object which can be RESOLVED using brightfield microscopy and the optics quoted. However, providing that the object could scatter light (a pit, for instance) much smaller objects could be OBSERVED, i. e., you could tell that they were there but nothing about size, shape, etc.
"Optimizing Light Microscopy..." has a detailed but easily understood discussion of the whys and wherefors. More info is available on our website: { {http://www.MME-Microscopy.com/education} . (Don't be scared away by the fact that the book was originally written for biologists and clinicians. There's lots of pertinent stuff for materials scientists, too.).
In July I asked this group for assistance in getting Leica to respond to us. I would like to thank all of you that responded and to let you know that I heard from them soon after my message was posted. I would also like to thank Ed Rae for resolving the problem. I also apologize to Leica for taking this long to let you all know that they helped us out.
Sincerely Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
We would like to have regular object in the background to track cell movement relative to. We have used polystyrene beads in the past, but these are very difficult to work with and expensive. Someone in materials science recommended Lycopodium powder which I gather is some kind of dried fungal spore. Does anyone know how large and/or regular these spores are? Any alternative suggestions? We are looking for 0.5-5um and (obviously) easily visualized. Thanks- Dave Knecht
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
For the first clue, look at the objectives. If they are all 160 mm (a marking on the objective barrel like 160/0.17), they can be interchanged. If there is a sign for "infinity" (a figure 8, on its side) instead of 160 on both, they can be interchanged. The only problem arises when one is "fixed tube length" (160mm) and the other is "infinity corrected".
Re: Confocal - better check with Olympus. The hardware fitting and the optical planes may or may not be compatible.
Dear Simon, } } Can anyone point me in the right direction concerning the preparation } of inorganic colloid samples in moderate/high ionic strength waters for } quantitative analysis by TEM? The dispersions contain relatively low } number concentrations of particles which we are trying to measure. } What gives the dispersion its high ionic strength, and do you want to analyse it? If you are only interested in the particles, and if the high IS is not from volatile compounds, you will have to rinse the particles in H2O (or some volatile buffer). If you want to analyse the liquid phase, you will have to get areas where it is on the grid, but no particles.
} So far, samples have been prepared by ultracentrifuging onto carbon } films (based on published method of Nomizu et al. Electron } microscopy of nanometer particles in freshwater, Anal. Chem. vol } 60, 2653-2656, 1988) and wicking the residual solution away with a } filter paper.
This seems to be a good start. If you do not wick away all of the solution, you could plunge-freeze the grid and analyse the frozen-hydrated specimen, then freeze-dry in situ and re-analyse. Do the first on Friday in a cryo-holder (which must be kept full somehow), then raise the temp to ~-90 C. Leave under vacuum for 24 hrs, raise temp to -80 C, leave for another 24 hrs, raise to -70 C, leave for 24 more hrs, then do the analysis.
} The problem is that there are considerable drying } artefacts such as matting of particles, uneven dispersion of } particles within a grid square (possibly due to surface tension } effects from residual water films during drying?) and } crystallisation of residual salts. } The cryo procedure should take care of all but the crystallization problem (unless the residual salts are volatile--then they will be gone too).
} I'd be most interested to hear from anyone with experience of } preparation of this type of sample or any good ideas. } No experience with this procedure, but it was featured in an EDS class I took -at- Lehigh. Possibly some of the Lehigh folks will also reply. Yours, Bill Tivol
Now that our TEM is fully functional I was hoping to be able to *do* something with it. During sample preparations we have typically used wax/heater to place samples to glass slides, etc. I would rather have an epoxy (acetone soluble) which would eliminate the need for heating during initial polishing. Any suggestions?
Also, we are looking for an epoxy (acetone INSOLUBLE) which can be used for the final specimen, for instance (especially), bonding two silicon wafers back to back before/after milling. Any suggestions here are greatly welcome. We aren't too picky about (non)conducting, just an epoxy which will be strong, adhere very well to silicon to a reasonable temperature range and not lose cohesion in acetone/ethanol, etc.
__ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3392 [x4] `___/ //\./=/~\====\ \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
Actually, purely from a basic physics point of view an LM is limited to the wavelength of the light used. YOu say you have a 540(nm) light source, which would be an arc lamp or some other source like a laser in order to acheive 540nm rather than a broad range of wavelength values. Especially since you have reported 0.4microns (400nm) as the smallest level of resolution. YOu would need to have 0.4micron (400nm, blue) light to truly be able to have resolution. The lower part of the *visible* range is around 0.25-0.3microns. Hence LM's are limited. EM's on the other hand make use of much smaller wavelengths (higher frequncies) of light to resolve far smaller images...down to angstroms (the size of atoms) (by higher frequencies it is meant that we have higher energies---E = hf.)
So yes it is a silly question, but even as a grad student I think it's good to "recall" these things. Sort of like picking up a history book.
__ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3392 [x4] `___/ //\./=/~\====\ \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
On Wed, 23 Sep 1998, Ingram, Mike wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This may be a silly question. What is the smallest object ( using } brightfield illumination) that can be observed. For example, I } calculated resolution of .4 micron for a 100x, .90 NA objective and } wavelengh value of 540. This is the smallest separation between } features that can be resolved. Is this also the smallest feature that } can be observed given a single feature on a smooth background? } } Michael Ingram } Rodel, Inc } Polishing Lab } 451 Bellevue Rd } Newark, DE 19713 } (302) 366-0500, ext.: 2545 }
Tim (TJ) LaFave Jr. wrote: ================================================ Also, we are looking for an epoxy (acetone INSOLUBLE) which can be used for the final specimen, for instance (especially), bonding two silicon wafers back to back before/after milling. Any suggestions here are greatly welcome. We aren't too picky about (non)conducting, just an epoxy which will be strong, adhere very well to silicon to a reasonable temperature range and not lose cohesion in acetone/ethanol, etc. ================================================= For this purpose, we have sold quite a bit of an interesting epoxy-phenolic adhesive called M-Bond™ 610 for this purpose. Technical information, including mixing and other use instructions can be found on our website below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The best material to use as an acetone soluble mounting material that you=
do not need to heat is a cyanoacrylate material (ie super glue). There have been many suggestions over the years as to the best including Sally Hansen's quick bonding nail glue, Loctite 460, and others. The Loctite product is available from a Loctite distributor and seems to be a good choice especially for Tripod Polishing. For less critical applications, any cyanoacrylate will do. They all vary in viscosity and curing time, b= ut generally if you get a good fresh product it will work. A good acetone soluble low temperature mounting wax is our QuickStick 135. This is a clear wax that flows at about 135 degrees C. The same material is also sold under the name Crystalbond.
As for non-acetone soluble epoxies, the 2 part epoxy that you want is mad= e by:
Epoxy Technology, Inc. 14 Fortune Drive Billerica, MA 01821
TEL: 800-227-2201 FAX: 978-663-9782
The product you want is EpoTek 353ND. They sell it in an 8 ounce kit whi= ch is the smallest size they offer. This is often used in TEM cross section= s.
Another material that is often used is M-Bond 610. That material is made=
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "Tim P. LaFave Jr." }
Now that our TEM is fully functional I was hoping to be able to *do* something with it. During sample preparations we have typically used wax/heater to place samples to glass slides, etc. I would rather have an epoxy (acetone soluble) which would eliminate the need for heating during= =
initial polishing. Any suggestions?
Also, we are looking for an epoxy (acetone INSOLUBLE) which can be used for the final specimen, for instance (especially), bonding two silicon wafers back to back before/after milling. Any suggestions here are greatl= y welcome. We aren't too picky about (non)conducting, just an epoxy which will be strong, adhere very well to silicon to a reasonable temperature range and=
not lose cohesion in acetone/ethanol, etc.
__ _-=3D=3D-=3D_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. =
Department of Physics \ \\ " / =
University of North Carolina, Charlotte } =3D\\_/` { Charlotte, NC 28223 ____ /=3D | \_|/ _' `\ _/=3D=3D=3D \___/ (704)547-3392 [x4] `___/ //\./=3D/~\=3D=3D=3D=3D\ \ // / | =3D=3D=3D: http://www.iit.edu/~lafatim | ._/_,__|_ =3D=3D: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /=3D=3D:-\ `.__' `-____/ |--|=3D=3D: Such that the future be theirs \ \ =3D=3D=3D\ :=3D=3D:`-= ' to shape and direct. _} \ =3D=3D=3D\ /=3D=3D/ -----------------------------------------------------------------
Thank You for all the answers given to my question about using Helium gas in the ESEM: I asked:
Stowe and Robinson report about reducing beam scattering in conventional Low Vacuum SEM's (Scanning, Vol. 20, 57-60). Are there any experiences using Helium in an ESEM from Electroscan or Philips with a special ESEM-detector? Is the ionization efficiency high enough to get a good performance for amplifying the electrons coming from the sample? How is the image quality compared to e.g. water vapor?
Kind regards
Rainer Ziel ------------------------------------------------------------------------ -------------------------- Roger Moretz wrote:
I can't give you the exact answers you are looking for, but hopefully the following can point you in the right direction. At the '99 MSA meeting in Atlanta, there was a session on ESEM. Several papers were given by the group from the Cavendish Laboratory, especially one on SEM at freezer temperatures by A.L. Fletcher, ... & A.M. McDonald. In an attempt to maintain the proper humidity without icing or thawing, this group used other gases, and determined that N2 was minimally acceptable in terms of obtaining desired imaging SEs. Discussion following the paper included comments on the scattering and SE generation that could be expected from lower atomic number gases, including He. I would suggest your contacting this group. Additionally, I would recommend that you do a literature search for the work of K. Rudiger-Peters and the various publications he has on the physical characteristics and SE generation in the ESEM. I think most of those papers were in the journal Scanning. The principal author on the ESEM is of course Danilatos, and his publications span from 1982 to 1990. Since I do not have ready access to those publications (everything is in boxes, no longer arranged as per my reference manager database) I can't give you figures or exact references even, but I believe that he did publish on different gases. Hope this helps.
Roger Moretz Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. ------------------------------------------------------------------------ ------------------------------------
Bradley L. Thiel wrote:
The question that you posted to the microscopy list server was passed on to me. We have investigated the imaging and amplification properties of several gases in the ESEM. Unfortunately, we have not been very efficient at publishing the results....
Helium actually works very well in the Philips-ElectroScan instruments. However, it is not a very efficient signal amplifying gas, so the emperical results can be a bit deceiving. First, because of the small elastic scattering, the probe-skirt formation is much reduced relative to water vapour. Second, the gas is even less efficient at amplifying the spurious signal from primary beam and backscattered electron ionisations. This means that although the signal collected by the ESD/GSED is not amplified much, it is a very pure secondary signal. The reduced skirt also gives stronger useful contrast.
Unlike other gases, He does not exhibit a peak in its amplification efficiency within the pressure range accessible in the ESEM. It is difficult maintaining pressures above about 8 Torr, because of backstreaming up the column. RP3 has a difficult time pumping the large volume of He.
Consequently, you should use a moderate pressure of gas, avoid detector biases that give arcing, and use the electronic signal amplifier and/or image integration to get a good image.
You may wish to see A.L. Fletcher, B.L. Thiel, and A.M. Donald, Journal of Physics D: Applied Physics, Vol. 30, 2249-2257 (1997). for some discussion of gases.
I hope this is of some help to you. Please feel free to contact me if you have other questions.
Best Wishes, Brad Thiel Polymers & Colloids Group Cavendish Laboratory Department of Physics University of Cambridge ------------------------------------------------------------------------ -------------------- Warren Straszheim wrote:
We routinely use He in our Hitachi 2460N, but we normally run at 40 Pa and use a backscattered electron detector. The resolution is _much_ better than air, and probably better than water vapor. I don't know how it would behave with the specialized sec. e detectors.
Also recall seeing at least one slide on the effect of gas type and pressure on scattering during ESEM sessions at the Atlanta MSA meeting. Sorry, I cannot remember who showed it.
Warren
------------------------------------------------------------------------ -------- Thank You
Rainer Ziel ------------------------------------------------------------- Dipl.-Phys. Rainer Ziel Akzo Nobel Central Research ACR-O/RMG-EM 63784 Obernburg
For an acetone soluble glue that's stronger/better than wax you need to use a cyanoacrylate; any of the family of superglues available in hardware stores. Look on the back of the tube. We use Ross Super Glue and Loctite 460 (preferred). Remember to get a cyanoacrylate *not* an epoxy. Time to dissolve in acetone isn't much different than wax.
For a glue that won't dissolve in acetone we use M-Bond 610 from Measurements Group, Inc., Raleigh, NC ph 919-365-3800, or obtainable from e.m. suppliers. This product is used to bond strain gauges to boilers, etc., but nothing beats it for TEM prep. Er, ah, if you call Measurements Group you may want to concoct a story about having a boiler that needs a strain gauge--better that than see the price go up if they tumble to the fact that the stuff is so good for TEM. :-) Despite their instructions to cure at over 150C for hours we routinely cure it at 70C for less than an hour. No problem at 30C, or even room temp, over night. But isn't 30C = room temp in NC?...never mind.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com IBM Analytical Services; http://www.chips.ibm.com/services/asg
This is a multi-part message in MIME format. --------------B9270C0DCC3F8B822F5E5C45 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Dear Tim,
If you wish to use a product which requires no heat for curing, try the LocTite 460. Depending on the amount of glue placed on the surface, it will harden in about 10 to 15 minutes. Larger mass will require longer cure times.
Every thin film epoxy, will require heating. Epoxies require mass to generate exotherm to cure and a thin film of epoxy does not generate enough heat to cure at RT. Even waxes will require heat, however if you mix the wax with acetone and let it stand, the acetone will evaporate and the wax will cure when this occurs. Time is dependent on the ratio of wax to acetone. M-Bond 610 also requires heat making it useless for your requirement.
We stock both LocTite 460, MBond 610 and EpoxyBond 110(similar to the Epoxy Technology epoxy). Should you have any further questions, please contact me at 800-675-1118 or via email.
Good Luck,
Gary Liechty Allied High Tech Products, Inc.
Tim P. LaFave Jr. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Now that our TEM is fully functional I was hoping to be able to *do* } something with it. During sample preparations we have typically used } wax/heater to place samples to glass slides, etc. I would rather have an } epoxy (acetone soluble) which would eliminate the need for heating during } initial polishing. Any } suggestions? } } Also, we are looking for an epoxy (acetone INSOLUBLE) which can be used } for the final specimen, for instance (especially), bonding two silicon } wafers back to back before/after milling. Any suggestions here are greatly } welcome. We } aren't too picky about (non)conducting, just an epoxy which will be } strong, adhere very well to silicon to a reasonable temperature range and } not lose cohesion in acetone/ethanol, etc. } } __ _-==-=_,-. } /--`' \_-at---at-.-- { } Tim (TJ) LaFave Jr. `--'\ \ {___/. } Department of Physics \ \\ " / } University of North Carolina, Charlotte } =\\_/` { } Charlotte, NC 28223 ____ /= | \_|/ } _' `\ _/=== \___/ } (704)547-3392 [x4] `___/ //\./=/~\====\ } \ // / | ===: } http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ } \/ \\ \\`--| / \\ } ---------- +*+ ---------- | _ \\: /==:-\ } `.__' `-____/ |--|==: } Such that the future be theirs \ \ ===\ :==:`-' } to shape and direct. _} \ ===\ /==/ } -----------------------------------------------------------------
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I use M-Bond 610, it comes in an adhesive kit form M-Line Accessories, Measurement Group Inc. Raleigh, NC. M-Bond is designed to mount strain gauges, say to evaluate stress and strain in airplane crashes. The epoxy is a two part, mixed, stored refrigerated, it has about a one month shelf life. Unmixed the two components refrigerated, have a shelf life of about a year. The mix is very thin, low centipoise, due to a high solvent content, this also allows the mix to move into very small spaces, via capillarity.
-- Respectfully, Bob ( Robert G. ) Lawrence Failure Analyst Motorola Phoenix Corporate Research Lab 2100 E. Elliot Rd. MD EL-703 Tempe, AZ 85284-1806 Phone: 602-413-5848 Fax: 602-413-4952 Pager: 1-800-759-7243 PIN 834-2458
We are seeking information on optics that could image the inner surface= of a circular object when shot from above. For example, what type of mirro= r could be placed in the inside of a waste-paper basket to photograph its inner= walls it from above? Our interest is in the simpliest type of mirror (e.g. hemispherical?) with the minimum moving parts.
Please reply off line to: jaincavo-at-goodyear.com Thank you for your help in this matter. =
As far as the acetone insoluble material, I used the LX112 resin kit from Fullam for 7 years (on Si thin sections) with great success. It is now called EPOX 812. Our ratios were as follows: LX112 resin = 5 grams NMA = 4.3 grams DMP - 30 = 7 drops This was well mixed and then the Si slices were "dunked" in the mixture, placed in a teflon vice and then baked at 85 degrees C for 12 hours. This results in a laminate that is resistant to just about everything. It is not as quick as some of the other materials but it is very reliable. Hope this helps.
John Staman Consulting FA Engineer LSI Logic, Colorado Springs
} -----Original Message----- } From: Tim P. LaFave Jr. [SMTP:lafatim-at-charlie.cns.iit.edu] } Sent: Wednesday, September 23, 1998 4:26 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Specimen epoxies for TEM } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Now that our TEM is fully functional I was hoping to be able to *do* } something with it. During sample preparations we have typically used } wax/heater to place samples to glass slides, etc. I would rather have } an } epoxy (acetone soluble) which would eliminate the need for heating } during } initial polishing. Any } suggestions? } } Also, we are looking for an epoxy (acetone INSOLUBLE) which can be } used } for the final specimen, for instance (especially), bonding two silicon } wafers back to back before/after milling. Any suggestions here are } greatly } welcome. We } aren't too picky about (non)conducting, just an epoxy which will be } strong, adhere very well to silicon to a reasonable temperature range } and } not lose cohesion in acetone/ethanol, etc. } } } } __ _-==-=_,-. } /--`' \_-at---at-.-- { } Tim (TJ) LaFave Jr. `--'\ \ {___/. } Department of Physics \ \\ " / } University of North Carolina, Charlotte } =\\_/` { } Charlotte, NC 28223 ____ /= | \_|/ } _' `\ _/=== \___/ } (704)547-3392 [x4] `___/ //\./=/~\====\ } \ // / | ===: } http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ } \/ \\ \\`--| / \\ } ---------- +*+ ---------- | _ \\: /==:-\ } `.__' `-____/ |--|==: } Such that the future be theirs \ \ ===\ :==:`-' } to shape and direct. _} \ ===\ /==/ } ----------------------------------------------------------------- }
We just look delivery of a new Philips XL30/TMP so the ETECs must go. Here's the deal. Three ETEC Autoscans. One is a split console/microscope flavor. One was working, one was semi-working. Two WDS detectors and WDS electronics. You pickup and take away (hint, need a lift gate truck). If no one wants these fine examples of american engineering from the '70 they will be scraped.
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
It is widely printed in every textbook to prepare gelatin subbed slides with an addition of chrom alum. Its oxidative properties are supposed to keep the gelatin from taking on bacterial populations. Bacteria are no problem if the gelatin solution is used, and discarded. However, there is a lot of grief with fading of sections, or color change of sections, when stained with the common LM stains. Fading of sections is a complicated problem, but one factor is the propensity of LM stains to react to redox shifts. Therefore, do not use chrome alum in subbing solutions. What if you use the slides for immunostaining and you have a nice underlayer of chrome alum below the section? Chrome alum makes perfect sense in the path lab perhaps. That is where it started - with paraffin sections. Many, many conceptual errors have been made by simply applying the methodologies designed for paraffin section to methods employed in TEM. Those two research specialties have very little in common. Hildy
Maleate buffers are not used for primary or secondary fixation. Why not use cacadylate? The maleate buffer system is very useful for enbloc use of UA in tissues which have been previously treated with osmium. They can be made to approximate the pH of UA in solution, causing the osmium to be stable and not move slightly. (this method can yield most beautiful membrances) If you want a reference on that, let me know. Why not use cacadylate buffer? It is toxic, but it would seem to be the method of choice if your specimens tolerate it. Make stock cocadylate at two times the strength. Wear a mask when weighing it. The powder is dangerous. Bye, Hildy
Does anyone know where Rapidoprint DD37E repair parts and chemicals might be obtained? Also, does anyone know if Kodak still makes Motion Picture #5302 is still made? TIA.
Chuck Butterick Engineered Carbons, Inc. Borger, Texas
Can anyone tell me a good way of removing the polymer packaging from an IC chip? I'd like to strip away the packaging and use the IC as an SEM specimen.
Many thanks, steve
-- Steven Kim | stevekim-at-nwu.edu MLSF, Room 3078 | 847.491.5888 (work) 2225 N. Campus Dr. | 847.491.7820 (fax) Evanston, IL 60208 | 847.332.1069 (home)
Dear All, In my reply to the Listserver of 9/17/98 on this subject, I mistakenly stated that Link had warmed the snout of their detector to prevent oil from settling on the detector. This, in fact, was the work of one individual on his Link detector, and did not work. The message I got from Link, privately, is as follows:
You recently sent a message to the microscopy listserver on the subject of oil contamination. The text was as follows:- ************* This is a problem of all SEMs, not just JEOL, and really is a result of the EDS detector being the coolest spot in the chamber. The cleanliness of the vacuum system just regulates how long the contamination will take to build up. Link solved this problem by warming the snout of their detector. This doesn't solve the problem of oil contamination, it just moves it away from the EDS detector. I agree woth Steve Chapman that the oil is from the rotary pump. *************
I should just like to correct a few points in this message.
The conditioning circuit fitted to the Oxford Link detectors DOES NOT warm the snout of the detector. The conditioner acts on the internal components of the detector, and cannot remove oil contamination. Warming the end of an oil contaminated detector by any means is likely to cause expensive damage to the window.
If the end of a detector becomes contaminated with oil, cleaning must only be attempted by a qualified engineer - the windows are fragile, and if cleaning is attempted in the wrong way the window could fail.
We would appreciate it if you could publish a note correcting these points, using your own words - we don't want people reporting window failures because they have been warming the ends of their detectors to remove oil!
Thanks
Chris Lay Customer Support Manager Oxford Instruments Microanalysis Group
*********************************************** Microanalysis Group, Oxford Instruments Halifax Road, High Wycombe Buckinghamshire, HP12 3SE, England Tel: +44 (0)1494 442255 Fax: +44 (0)1494 524129 URL http://www.oxford-instruments.com/ *********************************************** Needless to say, I was somewhat taken aback by the harsh tone of this message, and I was suprised to learn that customers cannot clean the window of a Link detector themselves. My experience with a Kevex Quantum window detector is that serious work with light elements must be preceeded by a careful solvent wash of the window, since the thinnest oil film will seriously absorb the lighter elements. Kevex have a Technical Bulletin (#21) showing this very graffically on SiO2, before and after cleaning, and complete instructions on cleaning. I always clean the detector, now, just before anyone does light element work. I have a Link detector, but a service call from a qualified Link engineer would cost airfare for 3000 miles plus time charges and supplies: a minimum of $3500 CAN. Sorry if I misled anyone.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
I have reacted epoxy-like material by slowly dropping boiling, red fuming nitric acid on the chip. Be careful! This acid is very reactive and will produce noxious/toxic fumes.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anyone tell me a good way of removing the polymer packaging from an IC chip? I'd like to strip away the packaging and use the IC as an SEM specimen.
Many thanks, steve
-- Steven Kim | stevekim-at-nwu.edu MLSF, Room 3078 | 847.491.5888 (work) 2225 N. Campus Dr. | 847.491.7820 (fax) Evanston, IL 60208 | 847.332.1069 (home)
via smtpd (for sparc5.microscopy.com [206.69.208.10]) with SMTP; 25 Sep 1998 13:20:16 UT Received: from ptag2 (ptag2.pt.cyanamid.com [141.173.51.2]) by igate.Cyanamid.COM (8.8.6/8.8.6) with ESMTP id JAA25089 for {Microscopy-at-MSA.Microscopy.com} ; Fri, 25 Sep 1998 09:19:54 -0400 (EDT) Received: from ccmail.pt.Cyanamid.COM by pt.Cyanamid.COM (PMDF V5.1-9 #28913) id {01J27VI3Y90W8X1M0W-at-pt.Cyanamid.COM} for Microscopy-at-MSA.Microscopy.com; Fri, 25 Sep 1998 09:20:09 EDT
For those of us in other fields with some curiosity, what is this word? I assume it's short for something. Is it just glue or does it have magical properties?
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
I need the manual for an older AO Interference scope. There is no model number on the unit, but I believe it is Series 7 or 9, and I believe the manual is:
I would appreciate some input on how to successfully perform antigen unmasking on formaldehyde fixed archived skin. I have experimented = with 3 commercial compounds and had only moderate success. Most of the problems seems to stem from the multiple layer nature of skin and the thickness of the tissue. I would ideally like to be able to perform this procedure on section upward of 25 =B5m and up to 100 =B5m. At = this thickness the tissue has a hard enough staying on the slide w/o being microwaved or steamed. I have tried the same compounds + protocols = with free floating sections, but the results were equally unimpressive.
I do realize that I may be forced to cut much thinner sections, = possibly around 10=B5m. Any input on successful unmasking procedures and the thickness of the tissue the procedure was performed on would be much appreciated.
Ramin Rahbari PARKE-DAVIS Pharmaceutical Research Pathology and Experimental Toxicology 2800 Plymouth Road Ann Arbor, MI 48105 Office (734) 622-3383 Fax (734) 622-5001 Ramin.Rahbari-at-WL.COM
If this is the typical "phenolic", black resin plastic, fuming nitric acid brought to approximately 90 degrees C will do the job. My suggestion is to put the part in some type of "basket" to facilitate easy removal and progress monitoring. Depending on the size of the part, it usually takes 5 - 8 minutes (most of the time) at that temperature.
John Staman Consulting FA Engineer LSI Logic, Colorado Springs
} -----Original Message----- } From: Steven Kim [SMTP:stevekim-at-casbah.acns.nwu.edu] } Sent: Thursday, September 24, 1998 4:38 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Solvent for IC Packaging? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Can anyone tell me a good way of removing the polymer packaging from } an IC } chip? I'd like to strip away the packaging and use the IC as an SEM } specimen. } } Many thanks, } steve } } -- } Steven Kim | stevekim-at-nwu.edu } MLSF, Room 3078 | 847.491.5888 (work) } 2225 N. Campus Dr. | 847.491.7820 (fax) } Evanston, IL 60208 | 847.332.1069 (home) } } http://pubweb.acns.nwu.edu/~stevekim } }
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Hello Steven,
There is a company in Hayward Ca, called EKC Technology that sells a product you are looking for. It is called EKC 270 and will remove the px from the device under heat in about 30 to 45 minutes, depending on the thickness.
Good Luck,
Gary Liechty Allied High Tech Products, Inc.
Steven Kim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Can anyone tell me a good way of removing the polymer packaging from an IC } chip? I'd like to strip away the packaging and use the IC as an SEM } specimen. } } Many thanks, } steve } } -- } Steven Kim | stevekim-at-nwu.edu } MLSF, Room 3078 | 847.491.5888 (work) } 2225 N. Campus Dr. | 847.491.7820 (fax) } Evanston, IL 60208 | 847.332.1069 (home) } } http://pubweb.acns.nwu.edu/~stevekim
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I have used boiling sulfuric acid to remove epoxy packaging. I usually cut off all the metal leads first. It can take all day depending on the thickness, but this has the advantage of not removing the aluminum traces (some of the time!)
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
I'm interested in making/buying an amplifier/pre-amp for specimen current measurements. Does anyone use a Stanford Research Systems SR570? If so, how do you like it? Any suggestions for getting the best performance from SC as an imaging technique?
Thanks in advance,
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
} go. Here's the deal. Three ETEC Autoscans. One is a split } console/microscope flavor. One was working, one was semi-working. Two WDS } detectors and WDS electronics. You pickup and take away (hint, need a lift } gate truck).
The ETECs have been taken.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
I want to thank all of you who take the time to help me out when I am up a creek, or about to embark on something retarded. You have saved me endless hours of grief with your sharing of knowledge and experience. In return I try to help where I can. (The refrences you want for the maleate enbloc UA I will find on Tuesday). Bye, Hildy
A few more caveats and questions - helium can certainly produce a dramatic difference in image quality and the amount of X-ray scatter, especially at lower kVs. We are using it a lot in CL imaging as well. However there are a couple of complications arising from its increased thermal conductivity relative to nitrogen or air. -
- A liquid nitrogen-cooled stage requires noticably more LN2 - so temperature at the specimen surface MAY be a little higher than in other gases, depending on the detector positions I suppose.
More importantly -2. Pressure readout from a gauge depending on a thermal effect, as in the Piranis used in most chamber pressure control systems, will be more in helium than nitrogen at the same pressure. There are graphs around. The diifference is not very much in the 0.01-0.1 torr range, but diverges very rapidly - from memory an indicated pressure of 3.5-4 torr corresponds to a true pressure of only about 1.3 torr. Which may affect the pressure range a gaseous amplification system thinks it is working at? (Ionisation potential of helium is about 25eV, cf 15eV for nitrogen...I dont know if that is enough to have a big effect on detector efficiency?) We only use BSE and CL, so I've no data on this, but for what its worth in the pressure range we use, we dont get the swamping background CL signal in helium that comes in at higher pressures in air.
3. Permeability -the EDS on our VP system has a Be window, and we havent had obvious problems resulting from helium. However somebody with an ultra-thin window EDS once mentioned that after using helium for leak testing, they thought some gas had found its way into the EDS detector. Not a good prospect, especially given the thermal properties. Does anyone have any more information or experience relating to this?
Sally Stowe
} Rainer Ziel wrote - } Thank You for all the answers given to my question about using Helium } gas in the ESEM: } I asked: } } Stowe and Robinson report about reducing beam scattering in conventional } Low Vacuum SEM's (Scanning, Vol. 20, 57-60). Are there any experiences } using Helium in an ESEM from Electroscan or Philips with a special } ESEM-detector? Is the ionization efficiency high enough to get a good } performance for amplifying the electrons coming from the sample? How is } the image quality compared to e.g. water vapor? } } Kind regards } } Rainer Ziel } ------------------------------------------------------------------------ } -------------------------- } Roger Moretz wrote: } } I can't give you the exact answers you are looking for, but hopefully } the following can point you in the right direction. At the '99 MSA } meeting in Atlanta, there was a session on ESEM. Several papers were } given by the group from the Cavendish Laboratory, especially one on SEM } at freezer temperatures by A.L. Fletcher, ... & A.M. McDonald. In an } attempt to maintain the proper humidity without icing or thawing, this } group used other gases, and determined that N2 was minimally acceptable } in terms of obtaining desired imaging SEs. Discussion following the } paper included comments on the scattering and SE generation that could } be expected from lower atomic number gases, including He. I would } suggest your contacting this group. Additionally, I would recommend } that you do a literature search for the work of K. Rudiger-Peters and } the various publications he has on the physical characteristics and SE } generation in the ESEM. I think most of those papers were in the } journal Scanning. The principal author on the ESEM is of course } Danilatos, and his publications span from 1982 to 1990. Since I do not } have ready access to those publications (everything is in boxes, no } longer arranged as per my reference manager database) I can't give you } figures or exact references even, but I believe that he did publish on } different gases. Hope this helps. } } Roger Moretz } Toxicology } Boehringer Ingelheim Pharmaceuticals, Inc. } ------------------------------------------------------------------------ } ------------------------------------ } } Bradley L. Thiel wrote: } } The question that you posted to the microscopy list server was passed on } to me. We have investigated the imaging and amplification properties of } several gases in the ESEM. Unfortunately, we have not been very } efficient } at publishing the results.... } } Helium actually works very well in the Philips-ElectroScan instruments. } However, it is not a very efficient signal amplifying gas, so the } emperical results can be a bit deceiving. First, because of the small } elastic scattering, the probe-skirt formation is much reduced relative } to } water vapour. Second, the gas is even less efficient at amplifying the } spurious signal from primary beam and backscattered electron } ionisations. } This means that although the signal collected by the ESD/GSED is not } amplified much, it is a very pure secondary signal. The reduced skirt } also gives stronger useful contrast. } } Unlike other gases, He does not exhibit a peak in its amplification } efficiency within the pressure range accessible in the ESEM. It is } difficult maintaining pressures above about 8 Torr, because of } backstreaming up the column. RP3 has a difficult time pumping the large } volume of He. } } Consequently, you should use a moderate pressure of gas, avoid detector } biases that give arcing, and use the electronic signal amplifier and/or } image integration to get a good image. } } You may wish to see } A.L. Fletcher, B.L. Thiel, and A.M. Donald, Journal of Physics D: } Applied } Physics, Vol. 30, 2249-2257 (1997). } for some discussion of gases. } } I hope this is of some help to you. Please feel free to contact me if } you } have other questions. } } Best Wishes, } Brad Thiel } Polymers & Colloids Group } Cavendish Laboratory } Department of Physics } University of Cambridge } ------------------------------------------------------------------------ } -------------------- } Warren Straszheim wrote: } } We routinely use He in our Hitachi 2460N, but we normally run at 40 Pa } and } use a backscattered electron detector. The resolution is _much_ better } than } air, and probably better than water vapor. I don't know how it would } behave } with the specialized sec. e detectors. } } Also recall seeing at least one slide on the effect of gas type and } pressure } on scattering during ESEM sessions at the Atlanta MSA meeting. Sorry, I } cannot remember who showed it. } } Warren } } ------------------------------------------------------------------------ } -------- } Thank You } } Rainer Ziel } ------------------------------------------------------------- } Dipl.-Phys. Rainer Ziel } Akzo Nobel Central Research } ACR-O/RMG-EM } 63784 Obernburg } } Tel: (06022) 81-2645 } Fax: (06022) 81-2896 } E-mail: Rainer.Ziel-at-AkzoNobel.com } } } } ---------------------------------------------------------------------- Dr Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: GPO Box 475 ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 (0)2 6249 2743 |Australian National Univ. FAX 61 (0)2 6249 4891 |Canberra, Australia 2601 http://online.anu.edu.au/EMU/home.htm
A while back (a year ago), I was told by AGFA that the DD3700 was being phased out and would no longer be serviced and that graded Rapitone paper P 1-4, ETC would no longer be available. We now use polycontrast resin coated paper, but most older users are quite unhappy with using the Kodak filter sets to get contrast. As a result, we are switching over to the Mohr-Pro 8 system and also doing more digital imaging. I have four AGFA DD3700's which are about to be surplused, along with many extra parts which I have been stock piling for years. If any one is interested in obtaining these items, let me know. Also, if anyone knows of a source for graded rapitone papers that are still good, let me know and maybe we can work out a deal. Does any one out there like their Mohr-Pro 8 system? Any long term issues, maintenance, paper, ETC to watch out for ? Thanks, Thomas A Baginski, Room G-230 Technical Coordinator for Microscopy Uniformed Services University of the Health Sciences 4301 Jones Bridge Road Bethesda, MD 20814-4799
Postdoctoral Position in Environmental Catalysis/HREM
A postdoctoral position is available at Northwestern University to work on Environmental Catalysis. The work will involve in-situ work using a unique UHV-HREM and gas treatment of various samples (see http://www.numis.nwu.edu for some information about the hardware). A strong background in HREM and crystallography is highly desirable, and experience in catalysis and/or surface science will be significant; extensive experience in other types of TEM are an alternative. To apply, send an email to L. D. Marks at ldm3-at-apollo.numis.nwu.edu including a CV (text only, no attachments) and the names of referees.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Dear Sally, } } 3. Permeability -the EDS on our VP system has a Be window, and we } havent had obvious problems resulting from helium. However somebody } with an ultra-thin window EDS once mentioned that after using helium } for leak testing, they thought some gas had found its way into the } EDS detector. Not a good prospect, especially given the thermal } properties. Does anyone have any more information or experience } relating to this? } When I was a grad student studying the scattering of protons on He, we had a target which was an evacuated, completely sealed thin glass sphere. The target was filled by immersing the sphere in a container filled with He. The He would diffuse through the sphere, which we would then use until sufficient He had diffused out. Since the glass was thick enough to be mechanically self-supporting, one can conclude that He will diffuse quite readily through many materials usually considered imper- meable. I'm not surprised that He can get through an ultra-thin window, but I would be surprised if it didn't also get through a Be window. I would also not expect obvious problems from the relatively small amount of He which got through the small area of an EDS window. Yours, Bill Tivol
Hi: If you have a complete Zeiss 109 or 10C for sale or if you know of any please contact me. It does not have to be in working condition. Thank you, Peter Jordan, EMSI 909 694-1839
To prevent tissues from schrinkage, EtOH 50% can be used instead of 70%. This is a very good fixative for general plant anatomy, it can be used as a preservation fluid as well: samples can be stored in it for years. Large specimens can be fixed. Fixation of mitotic figures and nuclear detail is poor... Fixation time for specimens 1cmx1cmx0.5cm: at least 24h. washing in EtOH 70% or 50%, depending on EtOH concentration used in the fixative. _______________
Chrom-acetic acid -------------------------- CrO3, 10% soln in dist. water: 2.5 - 10ml acetic acid (10%): 5 - 10ml add distilled water to obtain 100ml of fixative. Prepare just before use. Keep samples as small as possible. Fixation time: not longer than necessary (max. 24h), otherwise: maceration! Wash in running (or at least frequently changed) water, untill most of the yellow stain is removed (complete removal is difficult!). Fixation and washing in the dark to prevent the formation of chromates. Good preservation of nuclear detail and almost all structures (mitochondria and proplastids are frequently lost). Fixatives containing CrO3 are good mordants prior to staining with safranin. Staining with hematoxylins (f.e. Delafield's) is dfficult, but Fe-hematoxylins (Heidenhain's, Weigert's) can be used. Waste disposal of chrome is problematic: check with your local authorities! _______________
Bouin -------- This wel known fixative for animal histology can also be used for plant tissues.
Prepare just before use (there are conflicting opinions regarding the shelf life of the solution).
Fixation time: up to 3 days. Wash in EtOH 70% untill most of the yellow color is removed.
---------- } Van: Botanique1-at-aol.com-at-sparc5.microscopy.com } Aan: Microscopy-at-sparc5.microscopy.com } Onderwerp: Glandular trichomes } Datum: zondag 27 september 1998 7:01 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } }
} Hello all, } } I'm looking for a fixation recipe for herbaceous plants... I'm wanting to } look at glandular trichomes.... Any comments are welcome! } } Stefanie } ****************** } Stefanie Galgon } Department of Biology } Northern Arizona University } smg4-at-dana.ucc.nau.edu } Botanique1-at-aol.com }
Thank you to those who have replied. I do appreciate the help. However, I realize now, that in my hasty initial email, I left out ALL the details... silly me! Now that I have a moment, I'll say a little more....
For a few years now, I have been looking at the differences of concentrations of particular terpenes in Salvia officinalis. A main focus has been the differences in young and mature leaves. I now want to compare anatomy and morphology of peltate glandular trichomes (the terminology is so scattered in this area... peltate, sessile etc.) in young and mature leaves using TEM. Fun stuff, eh?
During my literature search, I came across quite a few SEM papers and recipes. I figured I would just go with one of the basics I am familiar with, but I thought shooting the question out to you guys wouldn't hurt.
Thanks again for the input.
Regards,
Stefanie ****************** Stefanie Galgon Department of Biology Northern Arizona University smg4-at-dana.ucc.nau.edu Botanique1-at-aol.com
Dear All, Can anybody let me know the recipe for twinning-jet-polish of Ag? Thanks in advance. Ke Han Center for Materials Science Los Alamos National Laborotory Los Alamos, New Mexico NM87545, USA Tel: 505-6650771 Fax: 505-6652992
I know two people looking for employment in EM/microscopy . One has managed an EM lab for many years - their lab is about to close. The other is a post-doc in Genetics with lots of microscopy experience.
Any help/leads would be greatly appreciated.
Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Ken: The following is for the window technique but it might work for jet polishing: aqueous potassium cyanide at less than 20 degrees C. voltage of about 8 V and current density of 1.8A/cm.sq. The sample must be rinsed in distilled water and used at once or stored in ethanol} Source: Desmond Kay editor. Pg. 403.
Jordi Marti ---------- } From: ke Han To: microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
A colleague has an EMITECH K850 CPD unit that appears to be malfunctioning.
1. Can any one supply us with information on a US distributor who might me able to help?
2. Can anyone help us diagnose the problem? The unit is 3 years old but has never been used. Upon presurization with CO2, there is a large gas leak originating from inside the unit. Upon removal of the back, the leak is seen to originate from what I am guessing is the overpressure/ heating override sensor. In other words, the leak is coming from a stainless steel or chromed sensor that has a copper CO2 line in one end and two electrical leads off the top (brown and yellow?--I forget exactly what the colors are). My guess is that this is the sensor that disables the heater when some pre-set pressure is exceeded, but I could be wrong. At any rate, the electrical leads on top of the sensor extend from a black plastic insert--the leak is between the black plastic insert and the SS surround. Does anyone know if this sensor is repairable?
TIA
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
Details of the October 9, 1998, Midwest Microscopy and Microanalysis Me= eting at Purdue University are now posted on the MSA webpage at http://msa.microscopy.com/. Follow the Local Affiliate Societies links= to find the agenda and maps. For further information, you may contact me = via e-mail. Thanks!
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064 =
{excerpt} {fontfamily} {param} Times {/param} Postdoctoral Position=20
An NIH-funded postdoctoral position is available immediately for structural studies of HIV-1 gp120. This is a joint project between Ken Roux and Ken Taylor.=A0 The project involves three dimensional epitope mapping and segmental flexibility analyses of gp120, gp160 and polymers there of in complex with ligands and monoclonal antibodies, using electron microscopy=A0 (negative staining and cryo-EM) and computational image analysis.=A0 Experience in electron microscopy and/or computational image analysis is essential. The project provides opportunities for development of innovative approaches to single particle microscopy and image analysis.=A0 State of the art microscopy facilities include a Philips CM300-FEG and a Philips CM120, each equipped with a fully computer controlled goniometer (CompuStage) for tomography and low dose cryoEM data collection.=A0 Computational facilities include a cluster of DEC Alpha compute servers and numerous Silicon Graphics workstations.=A0 Digitizing facilities include a Perkin-Elmer PDS 1010M densitometer.=A0 The labs and facilities are part of an extensive interdisciplinary Structural Biology program with expertise in X-ray crystallography, NMR, protein engineering, 3-D electron microscopy.=A0=20
We have a copy of your E mail on the Server, I believe that you have been given the address on our USA and UK Office.
However, from your detailed description your analysis of the problem would appear to be correct. This switches off heaters on an overpressure, which normally does not occur in normal use.
These are the seal units which can't be repaired in a local situation, and Emitech will need to return it to the original manufacturer.
However as part of our Service Support Policy, which gives you two years warranty on all parts, we would be pleased to extend this and send you an advanced replacement free of charge. We would be pleased if you could then return the old one.
Please advise delivery address.
Kind Regards,
David Robinson
In message {v04003a04b23548a2ce8b-at-[141.233.130.134]} , "wise-at-vaxa.cis.uwo sh.edu"-at-Sparc5.Microscopy.Com writes } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all, } } We want to integrate a focused ion beam gun into our large chamber SEM. Who } has experience } in doing this? } } Thanks in advance, Martin Klein } ___________________________ } } VisiTec Microtechnik GmbH } Karl-Marx-Str. 14 } D-23936 Grevesmuehlen } Germany } } Fon: +49-3881-790-49 } Fax: +49-3881-790-48 } email: mklein-at-visitec-em.de } web site: www.visitec-em.de } } } }
Please help me out on this, as its been many years since I have used permount and similar mterials.
A user is mounting ostracods in Cytoseal. Initially, she has beautiful mounts. After a few days of drying at room temperature, however, she finds lots of bubbles under her coverslip. Is this buffer/solvent leaching out of her sample? What can she do to solve this problem?
thanks in advance
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-8759 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
JEOL Geremany. Dr. W. Knoll 0049/8165-77346 0049/8165-77512 (fax)
-----Original Message----- } From: Dr P. Echlin {pe13-at-cus.cam.ac.uk} To: Martin Klein {Visitec-at-t-online.de} Cc: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
We are looking for image analysis software that will detect small area= s of color (say yellow on black) and automatically send a relay to a control= system when a colored area is detected? Thanks in advance for your attention = to this posting. Please reply off line to: jaincavo-at-goodyear.com =
Has anyone had experience in resolving Golgi with 100x fluroescence microscopy?
I am trying to discern the tubular-vesicular mechanisms of PC-12 cells through fluorescence microscopy and want to label Golgi with BODIPY ceramide, but fear they are too small to show up decently.
Appriciate the help!
-- Vickie A. Kimler, Ph.D. Assistant Professor of Biology and Allied Health Director, Cancer Research Facility Mercyhurst College 501 East 38th St. Erie, PA 16546 Voice: 814-824-2169 FAX: 814-824-2188 e-mail: {vkimler-at-mercyhurst.edu}
Ke Han asked "Dear All, Can anybody let me know the recipe for twinning-jet-polish of
Ag?"
Bernie Kestel used the following technique on South Bay 550-B single jet instrument with good results: 60 ml perchloric acid 460 ml ethyl alcohol 280 ml butyl alcohol 150 ml butyl cellosolve
Temperature: -20 degrees centigrade. Voltage: 220 V. Current: 70mA. with a thin plastic diaphram having a 1.4 mm center hole retaining the specimen in it's holder. In -situ, magnified viewing of the polishing and the unit's 300 volt capability were very helpful.
-- Dr. Roseann Csencsits Electron Microscopy Center Building 212/C217 Argonne National Laboratory 9700 South Cass Avenue Argonne, IL 60439-4838 Phone: (630) 252-4977 Fax: (630) 252-4798
You can add a drop of xylene to dissolve the mounting medium then expose the slide to a vacuum for a few seconds to remove the bubles. For more details see the above reference.
Jim Harper microfab-at-aol.com
In a message dated 98-09-29 15:38:23 EDT, you write:
{ { Please help me out on this, as its been many years since I have used permount and similar mterials.
A user is mounting ostracods in Cytoseal. Initially, she has beautiful mounts. After a few days of drying at room temperature, however, she finds lots of bubbles under her coverslip. Is this buffer/solvent leaching out of her sample? What can she do to solve this problem?
thanks in advance
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-8759 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/ } }
Discussion came up regarding the use of maleate buffers. Some people asked for a protocol. I will give you the reference, as writing it out would take a long time. This protocol is wonderful for the preservations of membranes, and great for preservation in general. It calls for GA,PAF prefix in phos buffer (or cac, if absolutely necessary), a one second rinse and then postfixation in veronal acetate buffered osmium. Veronal buffer cannot be used as a prefixation buffer, but the bacteriologist have used it for eons for good membrane preservation. I have left the material to fix overnight in osmium, if I had a structure of special interest which contained a lot of unsaturated lipid. Next the prep is washed with maleate of a certain pH in order not to dislocate the osmium. The material is then enbloc stained with UA in maleate. There are no huge shifts of pH here in this method. Acetone dehydration (saves lipids), PO intermediate and epoxy follows. All steps are done on ice except the last 100% acetone. All this was backed up at an EMSA meeting years ago by Janet Boyne of SF. Her talk appeared in the Bulletin.
Here is the reference. Williams MC. Conversion of lamellar body membranes into tubular myelin in alveoli of fetal rat lungs. J Cell Biol 1977; 72:260
If you try this method and like it, make up large quantities of the buffers and freeze them in aliquots. Bye, Hildy
I have inherited an old Ladd Sputter coater, the unit works, but the
film thickness is twice that indicated on the meter. This machine has a
density switch to enter the density of the material being sputtered, it is set to the correct density. I have been working with Ladd and they have been very helpful, but this unit is so old and they have moved several times.
I am looking for a schematic/ manual for a Poloron MK1 Film Thickness Monitor, it is used in a LADD Sputter Coater, model 30800 , the Ladd model number on the film thickness monitor is 30808. Any help would be greatly appreciated.
-- Respectfully, Bob ( Robert G. ) Lawrence Failure Analyst Motorola Phoenix Corporate Research Lab 2100 E. Elliot Rd. MD EL-703 Tempe, AZ 85284-1806 Phone: 602-413-5848 Fax: 602-413-4952 Pager: 1-800-759-7243 PIN 834-2458
Hello everyone; I am about to revisit in situ hybridization using DIG labelled probes. One thing I remember from the course I took was to NOT use Permount to mount the slides at the end of the protocol, or other organic based mounting media. My inclination is to use glycerol, but this doesn't seem to be a "permanent" treatment. Any suggestions would be greatly appreciated.
thanks in advance shea Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
I got no responses from my request of last week, so I am calling out again in the hope that someone on the list does indeed have this manual:
AO-Baker Interference Microscope, a Series 7 or 9 (I believe) built upon a basic AO Series 4 binocular scope, made probably in the late 50s or early 60s
Again, I will pay for copying costs and postage.
ALTERNATIVELY: I heard that someone in Canada has been collecting old microscope manuals for resale. Does anyone know who this person is?
Microscopy List Server {Microscopy-at-Sparc5.Microscopy.Com}
Dear Recipient,
I am hoping to reach email or internet catch-all addresses for microscopy secondhand equipment dealers.
Thieves stole three slide cabinets from our histology teaching lab over the last weekend. We bolted the stable door after the horse had flown, they came back yesterday lunch hour, forced the door, and took three more.
The cabinets must be valuable. The slides that they contained were in part invaluable.
I knew about the confocal list server, but I figure that confocal users will not be the class of person looking for antique slide filing cabinets. One of them was a rare [I guess late Victorian] beauty, mahogany, glass doors and ivory handles on all the 30 drawers. That was, of course, the one with our most valuable teaching slides in. The others were of a white coloured wood, probably beech.
I cannot imagine that the thieves would try to trade the slides because they would be too easily identified.
I would like to circulate to people who might be offerred these goods for sale and to ask them to contact us for further identification.
Yours sincerely,
Alan Boyde
Alan Boyde, Professor Hard Tissue Research Unit Department of Anatomy and Developmental Biology University College London Gower Street London WC1E 6BT United Kingdom Phone: If I do not answer on +44 171 419 3316 [or 3320/3322/3321] There are recording machines for messages up to 3 minutes on 3313 [and 3315] FAX +44 171 391 1302
On Tue, 29 Sep 1998 14:57:40 +0000 "Vickie A. Kimler, Ph.D." {vkimler-at-mercyhurst.edu} wrote:
} } } Has anyone had experience in resolving Golgi with 100x fluroescence } microscopy? } } I am trying to discern the tubular-vesicular mechanisms of PC-12 cells } through fluorescence microscopy and want to label Golgi with BODIPY } ceramide, but fear they are too small to show up decently. } } Appriciate the help! } } Vickie,
Its a piece of cake. We have undergrads do ceremide-Golgi labeling in lab exercises and they get beautiful pictures to take home and wow their parents, when recorded on a color video printer. One wrinkle that helps: Incubate the cells in ceremide for 10' at 4 deg C, then warm to 37. The dye will bind to the cell surfaces at 4 and be taken up synchronously and transferred to the Golgi when the cells are warmed.
-Dennis ---------------------- Dennis Goode Dept. of Biology University of Maryland College Park 20742 dg0-at-umail.umd.edu
} Microscopy List Server {Microscopy-at-Sparc5.Microscopy.Com} } } Dear Recipient, } } I am hoping to reach email or internet catch-all addresses for microscopy } secondhand equipment dealers.
To the list:
We all have sympathy when theft like this happens. Fear and loathing are two thoughts that come to mind.
Perhaps the following story will help a little.
Several years ago someone stole a computer that was a part of an imaging workstation. The poor soul whose lab was hit had his wits about him. He placed a classified ad in a local buy and swap newspaper (one that was known to list stolen merchandise from time to time). The ad read something like: "Desparate, looking for an older computer board with a serial number from between n and n+x. The board is usually mounted in a computer model XX. Will pay top dollar for the board." He gave his home phone number. He got a phone call within 24 hours. He actually got the caller to give him the serial number off of the computer. It was the lab computer. He arranged to go over to the caller's house to buy the board; of course, accompanied by a police detective. They recovered the computer and arrested two guys on the spot. They also found other "hot" items at the house. Sometimes you get lucky.
Blystone in Texas
-------------------------------- Robert V. Blystone, Ph.D. rblyston-at-trinity.edu
Department of Biology Trinity University 715 Stadium Drive San Antonio, Texas 78212 210.736-7243 FAX 210/736-7229
We would like to look at some heavier elements using the TEM, specifically alloys with Pt and Rh. Does anyone know the equation for calculating the thickness (thinness) requirements for samples to be electron transparent for different accelerating voltages?
TIA, John -- John Phelps NIST / DIV 853 Boulder, CO 80303
I need a TEM specimen consisting of gold monocrystalline islands on a thin, amorphous and preferably low atomic number supporting film for a study. I believe that a sample such as this is available commercially but am having difficulty locating a supplier. Many suppliers have standard specimens with polycrystalline gold islands evaporated onto a holey carbon film. However, I have examined this type of specimen and determined that it will not work for my study. Does anyone have any suggestions where I might find a supplier for a TEM specimen having monocrystalline gold islands with diameters of a few nanometers.
Thanks in advance.
_____________________________________________________ Michael Brickey (brickey-at-ecn.purdue.edu) Graduate Research Assistant School of Materials Science & Engineering Purdue University Office: MSEE S039 Phone: (765) 494-8751 _____________________________________________________
This won't be of much help for Mr. Boyde, but these are some of my guidelines when buying second-hand equipment... ---------------------------------------------------------------------------
Second-hand buyers should be very carefull in buying: as the prevention of theft is allmost impossible, the only other solution is to spoil the thieves' market...
It's good practice to check serial numbers with the manufacturer and/or agent when buying second-hand equipment, especialy when it comes very cheap...
When the seller don't allow you to take a note of the number(s): don't buy!
When an item doesn't carry a serial number or the manufacturer doesn't exist anymore: check the source! When the seller don't want to reveal his source: don't buy!
These investigations are perhaps time consuming, but for amateurs (I believe that most of the second-hand buyers are amateurs as I am) it's part of the fun!
Never buy "on the spot"! I always take a buyers option for a week. Sure, I know that the view of a beautiful instrument you've always wanted can be very tempting... But waiting a while and do some investigations can be rewarding: about 4 years ago, on a "flea-market", I took an option on a very cheap second-hand Zetopan microscope (15 000 Belgian francs, about US$ 430). The seller wouldn't reveal his source, but he said "it's from a deceased GP in..."a small village nearby. Some investigations in old and more recent telephone books learned me, that there were only 4 GP's in that village, so I took the telephone... It took me about 5 minutes to find the widow of the man... She said:" Sure, we have sold his equipment to that man, but there are still a few boxes of junk left. If you want them, you can have them, we want to sell the house, it has to be emptied...". The "few boxes of junk" contained amongst other things a full set of PL APO objectives for the Reichert microscope, and... a second microscope...: a very decent binocular Will-Wetzlar BX200 lab microscope... Waiting can be very rewarding indeed...
The manufacturer/agent can give very usefull information based on the serial number: precise model, year of production, availlability of spare parts, manuals etc... Somethimes he can even tell if the instrument was stolen...
Manufacturers should have a database with a list of all sold equipment, containing information on the whole life cycle of the item... Perhaps this is common practise, I don't know, but it should! I do know that this is the case for some other items (f.e.: Volvo has a central computer in Germany, with continuously updated information on all Volvos sold in Europe). A database of that kind could be linked on other information and become a very powerful instrument for internal QC, client relations etc... This can't be much of a problem in the age of computers...
Michael Brickey wrote: ================================================== I need a TEM specimen consisting of gold monocrystalline islands on a thin, amorphous and preferably low atomic number supporting film for a study. I believe that a sample such as this is available commercially but am having difficulty locating a supplier. Many suppliers have standard specimens with polycrystalline gold islands evaporated onto a holey carbon film. However, I have examined this type of specimen and determined that it will not work for my study. Does anyone have any suggestions where I might find a supplier for a TEM specimen having monocrystalline gold islands with diameters of a few nanometers. ==================================================== So far as I know, our product shown on URL http://www.2spi.com/catalog/standards/othertem17.html would meet the above description in your posting.
I don't believe we are your only source for this kind of product, some of the other major EM product suppliers have a similar (or maybe identical) kind of sample, or at least they have in the past.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Has anyone besides myself sputtered breakfast bars? No, I'm not starting a secret society, but, I have noticed a profound difference in the deposition of AuPd from the jellied center of the bar through to the delicious crunchy outer coating. We have no problem viewing the ultrastructure of the jelly center, that coats very well; as does the immediate interface with the cereal. When we go to analyze the cereal it is extremely unstable {charging}. Most likely this is due to the great difference in porosity between the tasty center and the delectable cereal, but if anyone has any other idea please get back to me before the samples are eaten. Yes an ESEM would do well, I guess, but we do not have one; and the field emitter is not used for "dirty" materials. I need some good basic suggestions that I may not have thought of .
Thanks,
John Grazul Rutgers University Electron Imaging Facility
This is a multi-part message in MIME format. --------------7A1DE8B357E8F2A69AF59121 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Earl Weltmer wrote:
} Hi All, } } Would any interested Third Party Maintenance organizations drop me an } e-mail including their location, area of expertise, main contact, etc. I } wish to form a clearinghouse for all TPMs to share technical } information, customer referral, etc. } } I have been in business as an SEM third party maintenance organization } in Southern California for over 15 years. Quite often I am asked to } recommend someone or repair equipment outside our geographical area. } } We have everything to gain. The ultimate winner is the customer. } } Earl Weltmer } } Scanservice Corporation } Tustin, CA
Hi,
For all interested parties, attached is a list of the Third Party Maintenance Organizations that have responded. I make no guarantees of their reputation but I am a believer in the free enterprise system. The stronger and more competent will remain in business especially in this limited field.
Evex Analytical, manufacture of x-ray microanalysis systems and digital = imaging systems also services and upgrades x-ray analyzers and detectors = manufactured by Edax, Kevex, Noran, and PGT.
Evex Aanalytical 857 State Road Princeton, NJ 08540 609-252-9192 service-at-evex.com
John, I think that the best approach for this is to calculate the extinction distances for a few reflections at the different accelerating voltages. The equation for extinction distance (see chapter 13 of Williams' and Carter's book or for the complete description, go to the Purple Peril - Chapter 4.6 of Electron Microscopy of Thin Crystals, Hirsch, Howie, Nicolson, Pashley, Whelan -Plenum) is
Eg = Pi * Vc cos(thetaB) / (Lambda * Fg)
where Vc is the volume of the unit cell, thetaB is the Bragg angle for the reflection, lambda is the electron wavelength, and Fg is the structure factor calcuated for the particular reflection, g responsible for the Bragg reflection.
You could put it in a spreadsheet for several voltages, reflections and compare for some common materials that you have a feeling for such as aluminum or silicon. The rule of thumb for usable diffraction contrast imaging is that the specimen should be less than 5*Eg in thickness.
Acc. voltage Lambda
---------- } From: John Phelps To: Microscopy ListServer
We would like to look at some heavier elements using the TEM, specifically alloys with Pt and Rh. Does anyone know the equation for calculating the thickness (thinness) requirements for samples to be electron transparent for different accelerating voltages?
TIA, John -- John Phelps NIST / DIV 853 Boulder, CO 80303
John, I think that the best approach for this is to calculate the extinction distances for a few reflections at the different accelerating voltages. The equation for extinction distance (see chapter 13 of Williams' and Carter's book or for the complete description, go to the Purple Peril - Chapter 4.6 of Electron Microscopy of Thin Crystals, Hirsch, Howie, Nicholson, Pashley, Whelan -Plenum) is
Eg = Pi * Vc cos(thetaB) / (Lambda * Fg)
where Vc is the volume of the unit cell, thetaB is the Bragg angle for the reflection, lambda is the electron wavelength, and Fg is the structure factor calculated for the particular reflection, g responsible for the Bragg reflection.
You could put it in a spreadsheet for several voltages, reflections and compare for some common materials that you have a feeling for such as aluminum or silicon. The rule of thumb for usable diffraction contrast imaging is that the specimen should be less than 5*Eg in thickness.
We would like to look at some heavier elements using the TEM, specifically alloys with Pt and Rh. Does anyone know the equation for calculating the thickness (thinness) requirements for samples to be electron transparent for different accelerating voltages?
TIA, John -- John Phelps NIST / DIV 853 Boulder, CO 80303
You might use one of the Monte Carlo programs. Although I have not tried it, from what I have read "Electron Flight Simulator" would model your analysis. Try: http://www.small-world.net
We would like to look at some heavier elements using the TEM, specifically alloys with Pt and Rh. Does anyone know the equation for calculating the thickness (thinness) requirements for samples to be electron transparent for different accelerating voltages?
TIA, John -- John Phelps NIST / DIV 853 Boulder, CO 80303
I will soon be submitting a paper to a yet to be determined materials science journal (Acta Met., Phil. Mag. or Materials Science and Engineering A) and I need to prepare some SEM and TEM images. The methods that we have been used to produce these are quite tedious, and I am seeking simpler methods. For TEM images, we normally produce a print of the negative, mark it up with the suitable labels, take another photograph of it, and then have this printed in the correct proportions to fit into the paper. We also have in our lab a light table and MTI 70 series digital cameras, which can directly capture images to a Macintosh (using NIH Image). Is it acceptible to submit such digitized images to microscopy journals? Also, what sort of resolution is required? Also this is the first time that we will publish the SEM micrographs, taken on 4X5 Polaroid films. What methods are needed to insert these into papers? As far as formatting text with the images, I usually use MS Word, but I have access to other Linux and Unix packages as well. If anyone has experience with such issues, I would greatly appreciate any feedback, either on the list or by e-mail.
Thank you,
Kevin Croat Physics Dept. Washington University in St. Louis tkc-at-howdy.wustl.edu
For immediate distribution: ELECTRON MICROSCOPIST POSITION Department of Pathology, Yale University School of Medicine
RESEARCH ELECTRON MICROSCOPIST, to begin immediately. Full-time Research Associate, for NIH-funded investigation of the mechanisms of bile formation by the liver. Ultrastructural studies focus on mechanisms for hepatic secretion of phospholipids and cholesterol into bile, and how these mechanisms go awry in liver disease. Studies in progress have the potential to provide major insights into cholesterol homeostasis, since bile is the only effective route for elimination of excess cholesterol from the body. These studies are thus relevant to the human conditions of atherosclerosis, cholesterol gallstone disease, and jaundice. The research associate will play a major role in all aspects of the project as skills permit, including but not limited to: ultrarapid in situ cryofixation, tissue processing for TEM, transmission electron microscopy, digital imaging morphometry, small animal surgery, photography and computer skills, literature search skills, and administration of the laboratory effort. Additional optional skills include reports and presentations, tissue culture experimentation, and electron energy loss spectroscopy (EELS) with electron spectroscopic imaging (ESI). Education/work experience requirements: B.S. or B.A. degree with extensive electron microscopy experience preferred, or M.S. degree with training in electron microscopy. Yale University is an equal opportunity employer. Contact: James M. Crawford, M.D., Ph.D., Department of Pathology, Yale University School of Medicine, 310 Cedar Street, P.O. Box 208023, New Haven, CT 06520-8023. Fax 203-737-1064. E-mail {jamesmac.crawford-at-yale.edu} .
Thank you.
James M. Crawford, M.D., Ph.D. Department of Pathology Yale University School of Medicine P.O. Box 208023, 310 Cedar Street New Haven, CT 06520-8023 Tel 203-785-2784 Fax 203-737-1064 jamesmac.crawford-at-yale.edu
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These darn food scientist types are always throwing a cereal or two in our direction. The flaky outer coating is great for crunch but lousy for conductivity. About the only thing you can do is coat in presence of Argon to get the best coat possible and then use very low KV to reduce charging. A sample such as this is very low density and will not withstand high magnification (empty magnification is usually only a few thousand times in this kind of sample). Since you can only gather useful information at fairly low mags, you should have plenty of signal at 4-5KV even with conventional tungsten hairpin filament and standard gun configuration. We do this routinely with our JEOL JSM-840.....in fact we rarely do secondary electron imaging above 5-6kv on low density samples typical of life science material.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------
Fellow Sputterers,
Has anyone besides myself sputtered breakfast bars? No, I'm not starting a secret society, but, I have noticed a profound difference in the deposition of AuPd from the jellied center of the bar through to the delicious crunchy outer coating. We have no problem viewing the ultrastructure of the jelly center, that coats very well; as does the immediate interface with the cereal. When we go to analyze the cereal it is extremely unstable {charging}. Most likely this is due to the great difference in porosity between the tasty center and the delectable cereal, but if anyone has any other idea please get back to me before the samples are eaten. Yes an ESEM would do well, I guess, but we do not have one; and the field emitter is not used for "dirty" materials. I need some good basic suggestions that I may not have thought of .
Thanks,
John Grazul Rutgers University Electron Imaging Facility
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Kevin,
Being an old-fashioned guy with a limited budget, I have gravitated to a part low-tech, part high-tech approach that offers the path of least resistance (and expense) for me to get from microscope to manuscript. Using the darkroom, I print individual photos from negative to the size I want for the journal (usually 2" or 3" by 3" or 4"). Then I trim and miter multiple photos, mount them on heavy card stock (heat press or spray adhesive will do), and annotate with press-on letters. Those plates become the master, "engraver's" copy. I next have color photocopies made of the masters. Photocopying technology is pretty amazing these days--copies made on a color photocopier are at least as good as your photos are likely to look when they eventually appear in the journal. The photocopies are used for reviewing purposes. They cost about $1.50 each but are worth all of the darkroom time saved in making second generation prints (plus they reduce mailing costs and ship better). This way, I invest in the low-tech hardware (a darkroom and a pair of scissors) and pay somebody else ~$30/pub to stay current with the high tech photocopying hardware. Another possibility would be to scan photo or negative into computer, make up plates and annotate (using Photoshop or others), transfer to disc, and take disc to an outlet (Kinko's?) that has a photo-quality printer. This route would probably take as much time and cost as much as option #1, above. Also, image resolution could suffer greatly depending on your computer. The techno-jocks and computer sales people will recommend digital image acquisition, then cut, paste and annotate on computer, then output to a photo-quality printer. All that is wonderful, but I don't have the 10^nth dollars to invest in hardware and software--especially since that hard/software is obsolete within years, or months, of purchase.
Bob
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Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
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Have you considered making a replica? You will lose the ability to do elemental analysis and it won,t be as tasty, but they work well for this type of sample. Ref: Sheffield E. etal. American Fren Journal 81 (4): 128-133 (1991) } } Fellow Sputterers, } } Has anyone besides myself sputtered breakfast bars? No, I'm not } starting a secret society, but, I have noticed a profound } difference in the deposition of AuPd from the jellied center of the } bar through to the delicious crunchy outer coating. We have } no problem viewing the ultrastructure of the jelly center, that } coats very well; as does the immediate interface with the } cereal. When we go to analyze the cereal it is extremely } unstable {charging}. Most likely this is due to the great difference } in porosity between the tasty center and the delectable cereal, but } if anyone has any other idea please get back to me before the samples } are eaten. Yes an ESEM would do well, I guess, but we do not have } one; and the field emitter is not used for "dirty" materials. I } need some good basic suggestions that I may not have thought of . } } Thanks, } } } John Grazul } Rutgers University } Electron Imaging Facility
William R.McManus Supervisor Electron Microscope Facility Department of Biology Utah State University Logan UT 84322-5305
Michael; This is not too silly a question, because many people apparently get so caught up in worrying about resolution that they miss the difference between
resolution and visibility. You may be able to detect a particle much less than the resolution limit, but you just won't be able to tell anything about it. One example: the human eye can resolve about 100 microns-that means two particles less than 100 microns apart will appear as one! You can easily see a 20 micron hole in an aperture strip if you shine a light behind the aperture strip, but if there happen to be two 20 micron holes 25 microns apart (center to center) it will look about the same. Astronomers know this well. Most "stars" you see with the naked eye are really clusters of stars. You can see the single point of light (on a clear night) but you cannot tell how many stars are in the cluster without better optics that can resolve them. In general, features much smaller than your resolution limit CAN be detected, but only if the contrast is strong, and remember that detection and resolution are not the same.
John Mardinly Intel Subj: LM --- Ultimate Resolution Question
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This may be a silly question. What is the smallest object ( using brightfield illumination) that can be observed. For example, I calculated resolution of .4 micron for a 100x, .90 NA objective and wavelengh value of 540. This is the smallest separation between features that can be resolved. Is this also the smallest feature that can be observed given a single feature on a smooth background?
Michael Ingram Rodel, Inc Polishing Lab 451 Bellevue Rd Newark, DE 19713 (302) 366-0500, ext.: 2545
Hi Kevin, Should you wish your image reproduced at the hightest possible quality, I have several thoughts to present. While not to be too basic, we all know that with offset printing, the ink is carried to the paper with a metal plate - and the image is burned into the plate through a negative. And that this negative is produced completely by the publisher/printer. You should provide your source image (more follows) in its first and purest form - you should not size it, and you should certainly not run it through a computer for page placement, etc. The reason being that each of these steps will reduce final resoultion. Then, your publisher, printer or film separator will take your image, and with one pass put in on the film which will finally produce the metal plate - with very high quality photographic equipment. Sure, with this process in page make-up, you will have to leave space with the correct width-height ratio for your image. Others might prefer using a software system such as Pagemaker or Quark Xpress and make up the full page - including the image - to produice this final film.. OK - but the image will be somewhat degraded. As to source image, many have different opinions. Mine first would be 4 x 5 film, followed by 35 mm slides. Next, and close, would be digital. It is factual that with discs, occasionally bits are "dropped" and errors made - and without other advantage, I question its real value. And - disc processing tends to be a bit more expensive over all others. Last would be photos. I hope that your find these opinions of interest. Don Grimes, Microscopy Today
On Thu, 1 Oct 1998 wise-at-vaxa.cis.uwosh.edu-at-sparc5.microscopy.com wrote:
} Using the darkroom, I print individual photos from negative to the } size I want for the journal (usually 2" or 3" by 3" or 4"). Then I trim } and miter multiple photos, mount them on heavy card stock (heat press or } spray adhesive will do), and annotate with press-on letters. Those plates } become the master, "engraver's" copy. I next have color photocopies made } of the masters.
This is pretty much what we do as well most of the time.
snip
} Another possibility would be to scan photo or negative into } computer, make up plates and annotate (using Photoshop or others), transfer } to disc, and take disc to an outlet (Kinko's?) that has a photo-quality } printer. This route would probably take as much time and cost as much as } option #1, above. Also, image resolution could suffer greatly depending on } your computer. snip } All that is wonderful, but I don't have the } 10^nth dollars to invest in hardware and software--especially since that } hard/software is obsolete within years, or months, of purchase.
More and more, especially with color, we go this way. Also, many of our illustrations (not the micrographs) are computer generated anyway. The investment is not terrible, resolution is quite adequate and obsolesence is not a problem; just keep using it if it works and ignore the newer stuff.
I am a retired mechanical engineer with an interest in astronomy and = optics. I have recently acquired two old microscopes: 1. A B&L Metallographic microscope 2. A Richert inverted polarizing microscope I am primarily interested in studying meteorites (as a hobby) so the = metallographic scope can be used for iron meteorites and the polarizing = scope for thin sections of stones. The problem is that neither = instrument is complete and I have no documentation on either. Can someone give me a source of=20 documents (instructions, servicing details, or drawings) on these = scopes? Thanks. John R. Chappell 6111 E. Sunnyside Rd. Idaho Falls, ID 83406 hbjrc-at-srv.net
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#fff8e0} {DIV} {FONT color=3D#000000 size=3D2} I am a retired mechanical engineer = with an=20 interest in astronomy and optics. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} I have recently acquired two old=20 microscopes: {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} 1. A B&L Metallographic=20 microscope {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} 2. A Richert inverted = polarizing=20 microscope {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} I am primarily interested in = studying meteorites=20 (as a hobby) so the metallographic scope can be used for iron meteorites = and the=20 polarizing scope for thin sections of stones. The problem is that = neither=20 instrument is complete {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} and I have no documentation on = either. Can=20 someone give me a source of {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} documents (instructions, servicing = details, or=20 drawings) on these scopes? {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Thanks. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} John R. Chappell {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} 6111 E. Sunnyside Rd. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Idaho Falls, ID 83406 {/FONT} {/DIV} {DIV} {FONT size=3D2} {A = href=3D"mailto:hbjrc-at-srv.net"} hbjrc-at-srv.net {/A} {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {/BODY} {/HTML}
Hi All, I enclose details of a vacancy in our department. Any one interested please reply directly to Amanda or our administrator.
Ron
**************************************************************************** Department of Materials, University of Oxford
Postdoctoral Research Position, 3 Years from October 1998
Advanced Information Storage films with magnetoresistive properties
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A three-year post-doctoral research assistantship funded by the Advanced Magnetics Programme of the EPSRC is available in the Department of Materials at Oxford from October 1998 (or as soon as possible thereafter). The aim of the project is to assess the effects of microstructure on magnetic and transport properties in magnetoresistive spin-valve and spin tunnel-junction materials. The project is a collaboration involving Oxford, Cambridge and Plymouth Universities.
The project will primarily involve the characterisation of both tunnel-junction and spin-valve material using electron microscopy (EM) techniques. Films will be grown, their magnetisation reversal mechanisms will be studied using Lorentz EM and the microstructure and chemical composition using HREM and microanalysis. Lithographic patterning of the material will also be carried out.
EM experience is essential, preferably at an advanced level, and a knowledge of thin layered films or magnetic materials would be helpful, as would experience in lithographic processing.
Applications including a cv, list of publications, and the names and addresses of three referees should be sent to: The Administrator, Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH, UK, from whom further particulars are available. Please quote ref: AKPL. Further particulars are also available by e-mail: amanda.petford-long-at-materials.ox.ac.uk
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meeting point for developers and users working in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
I recently attached a list of the Third Party Maintenance Organizations (IBM PC,EXCEL format). Many of you e-mailed me and asked that I re-submit the list via regular e-mail.
Below is that list:
Company Contact Type Specialty Location Telephone e-mail Equipment Advanced Allen Research SystemsSampson Any Any Chicago 630.513-7093 ars-at-mcs.net
AMTEC Sam TEM Phillips New amtec-at-neca.com Amtower England
E-MAC Walter Probe Cameca Texas 281.879-7211 Corvos-at-aol.com Protheroe
ELECTROVAC TECH.Peter SEM ISI/Topcon Canada 905.294-9259 petere-at-pathcom.com Earl Focused Larry Resolutions Barry SEM ISI/Topcon Mass 978.689-3977 Focres-at-aol.com Micro-AnalyticalJoe Service Barney Probe Cameca Penn 717.299-0599 jbarney_microanalytical-at-email.msn.com
Quality Images Ken SEM ETEC Penn 717.456-5491 qualityimages-at-netrax.net Converse
Scanners Corp. Gary SEM Any Maryland 410.456-5491 qeaston-at-ibm.net Easton Scanservice Earl Corp. Weltmer SEM Any California714.573-9158 earlw-at-pacbell.net Scientific Inst.Alex Services Greene TEM Any Texas 512.282-5507 ablue-at-io.com
VEC Craig SEM Hitachi Colorado 303.689-2224 Franklin-at-idcomm.com Franklin Vitaly Feingold Any Any Georgia 770.232-1791 vitalylazar-at-worldnet.att.net Chuck Humphrey SEM ISI/Topcon 888.793-8103 cchumph-at-ibm.net
{P} I recently attached a list of the Third Party Maintenance Organizations (IBM PC,EXCEL format). Many of you e-mailed me and asked that I re-submit the list via regular e-mail.
{P} Below is that list: {TABLE BORDER COLS=7 WIDTH="100%" } {TR} {TD} Company {/TD}
I would like to get some information/references about 'Etching LR White for immunogold labeling'. I am recently working on labeling a cytoplasmic protein in the plant of Brassica anther/pollen. Few labels occur when I use the "regular" concentration of antibody. As I increase the antibody concentration, the background is getting higher and higher but still, not too much labels inside the cytoplasm. I wonder whether it is the problem of resin (LR White). Many thanks in advance.
Zhaojie Zhang Dept. of Botany and Microbiology University of Oklahoma Norman, OK 73019
A RESEARCH TECHNICIAN position is available in the laboratory of Tom Misteli at the National Cancer Institute, NIH, Bethesda, MD. The laboratory uses molecular biology tools in conjunction with microscopy techniques (confocal, live cell, deconvolution) to study the architecture of the mammalian cell nucleus and the interphase organization of chromosomes.
Candidates must have experience in fluorescence microscopy and in molecular biology techniques. Knowledge of cytogenetic techniques (FISH, chromosome painting, CGH) is an advantage. The position involves experimental work on ongoing projects, microscope maintenance and laboratory management. Development of independent projects will be encouraged.
This permanent position is funded by the NIH and includes NIH benefits. Salary is commensurate with experience. Starting date is early 1999. Send CV including a list of skills and names of three references to Tom Misteli, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724 USA, email: misteli-at-cshl.org.
Cold Spring Harbor Laboratory Cold Spring Harbor, NY 11724 T: 516 367-8478 F: 516 367-8876 misteli-at-cshl.org
Applications are invited for a POST-DOCTORAL POSITION in the group of Tom Misteli at the National Cancer Institute, NIH, Bethesda, MD. The laboratory studies the molecular mechanisms which control nuclear architecture and function in-vivo, particularly the coordination of transcription and pre-mRNA splicing (see Nature, 1997, 387, p.523, Science, 1997, 276, p.1495, Curr. Opi. Cell Biology, 1998, 10, p. 323). In addition, we are analyzing the spatial organization of genes and chromosomes within the interphase cell nucleus. We use conventional and time-lapse fluorescence microscopy of living cells in conjunction with molecular methods to address these cell biological aspects of gene expression. The laboratory is closely associated with a new NCI imaging facility equipped for laser scanning confocal microscopy, deconvolution methods, in-vivo microscopy, 4D-image analysis and multi-color cytogenetics.
Candidates must have experience in fluorescence microscopy and molecular biology techniques. Knowledge of cytogenetic techniques (FISH, chromosome painting etc.) is an advantage.
The NCI provides outstanding resources and a large, interactive environment to perform high level innovative research. The successful applicant will have the opportunity to develop his/her own projects within a highly interactive team.
This NIH funded position is available from January 1999. Please send CV including a list of publications and names of three referees to Tom Misteli, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724 USA, email: misteli-at-cshl.org
Cold Spring Harbor Laboratory Cold Spring Harbor, NY 11724 T: 516 367-8478 F: 516 367-8876 misteli-at-cshl.org
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager of Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
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Kevin, It appears that all your images are originally produced on film. This is great because a good negative taken to the darkroom and printed appropriately will normally result in the sharpest final image. Drawbacks are that digital images can be processed using a program like Adobe PhotoShop in ways that are difficult to do in the darkroom.
I don't want to get into the ethics of image processing but removing dust specks and adjusting contrast and brightness are probably OK with most people. It is a lot easier to "dodge" and "burn" on a computer than in the darkroom once you get the hang of doing it. However, final digital images, even those printed out on a top-of-the-line dye sublimation printer have a softness to them that darkroom prints won't have. You can run a sharpening filter on the images prior to printing but....that's where there will be disagreements as to how much and what type of filtering is ethically OK. Although we do digitally acquire SEM images in our lab, numerous tests have shown that normally the best images for future publication are obtained using film. I just did a test series while making demo slides for my SEM class, using a piece of fabric with fibers going in different directions. The digitization was very apparent in some fibers where you could actually see stepwise edges along fibers angled in certain directions to the scan ( digitized at 1280x960). The polaroid negative gave clean edges. It was even preferable when the negative was later scanned into the computer at 300dpi ...although I normally scan negatives in at at least 600dpi to pick up maximum detail.
I personally do a combination of things to get final pictures. If the plate is complicated, with many different images, I will often scan the negatives and then assemble the plate on the computer. I can resize and crop images at will so can easily play with the layout to get exactly what I want for publication. If a very good printer is available, you can label these "mock-ups" and use the result often as reviewer's copies or internal prints for co-authors.
Once I have my layout, I can go to the darkroom and in one printing, print the images to the sizes needed, matching contrast with adjacent images as I go. Mounting and labeling will then result in a top quality final plate. This plate is the one that goes to the journal for reproduction. We have 4x5 negatives made of each final plate. Excellent reviewer's copies can then be made from these negatives.
With an easy layout, I may not need the computer step and can do final images the first trip into the darkroom or can make some quick working prints, crop, etc. and then go back into the darkroom for final printing.
I have published digitally generated images when I had to do processing that could not easily be done in the darkroom. This is OK as long as you describe in detail what was done in the material and methods section of the paper. However, in my opinion, the bottom line is that regardless of what goes into the computer, if you cannot generate top quality output, this is not the method of choice for publication images. And reviewers must see good images in order to adequately evaluate a manuscript. Digitized images, although getting better all the time, are still not the same quality of good darkroom prints from good negatives.
Okey, off the soapbox and back to work..
Debby ------------------------------------------------- Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Greetings,
I will soon be submitting a paper to a yet to be determined materials science journal (Acta Met., Phil. Mag. or Materials Science and Engineering A) and I need to prepare some SEM and TEM images. The methods that we have been used to produce these are quite tedious, and I am seeking simpler methods. For TEM images, we normally produce a print of the negative, mark it up with the suitable labels, take another photograph of it, and then have this printed in the correct proportions to fit into the paper. We also have in our lab a light table and MTI 70 series digital cameras, which can directly capture images to a Macintosh (using NIH Image). Is it acceptible to submit such digitized images to microscopy journals? Also, what sort of resolution is required? Also this is the first time that we will publish the SEM micrographs, taken on 4X5 Polaroid films. What methods are needed to insert these into papers? As far as formatting text with the images, I usually use MS Word, but I have access to other Linux and Unix packages as well. If anyone has experience with such issues, I would greatly appreciate any feedback, either on the list or by e-mail.
Thank you,
Kevin Croat Physics Dept. Washington University in St. Louis tkc-at-howdy.wustl.edu
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When we export an image using TIFF format on our XL30/TMP, the image is compressed in the y-axis (or expanded in the x-axis). Screen and video prints are fine. This is under Windows 3.11 and version 5.39 of the XL30 software. Has any else see this problem or is it particular to our SEM? FEI/Philips has yet to respond to this problem.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
When we are using external scan, we eventually blow the filiment because the XL30's software increases the filament bias to max (6). Has anyone else seen this problem or is it particular to our scope? This is under Windows 3.11 and version 5.39 of the XL30 software. FEI/Philips has yet to respond to this problem.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
I recently started an SEM/TEM independent study of C. ellegans. I am runn= ing into some problems with the fixation methods and protocols for manipulating the worms. I could use any help that is out there! We started with glutaraldeyde and osmium solutions, but I=B9m using very old protocols and = I think they might need to be refreshed. =20
Thanks, Rob Reff
Research Student Under the Direction of:=20 Professor William J. Perreault Lawrence University Appleton WI 54915 (920) 730-8236
I believe we had the same issue with our Zeiss/Leo. The Tiff specification allows for non-square pixels, however most external image processing programs assume the pixels are square. I believe the SEM is outputing a file with rectangular pixels but other programs are ignoring it.
Lynn Rathbun
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Dr. Lynn Rathbun,Cornell Nanofabrication Facilty, Ithaca, New York Rathbun-at-cnf.cornell.edu Webmaster -at- Christian World Adoption www.cwa.org Webmaster -at- Joint Council on International Children's Services www.jcics.org (All opinions or representations of fact re: adoption are my own however)
I do not have the manual but have the catalog writeup in the van waters and rogers scientific catalog, it is brief but would that help any at all? thanks ed sharpe, archivist smecc
Dear Sirs, I am Nelson Fava and I work with a CAMEBAX SX50 at the U. de Brasília, Brazil (SX#359). HOw are you? I am looking for a used camebax or a used JEOL microprobe to buy. Please contact me if you have any news
MME offers customized on-site courses in all areas of microscopy which incorporate both information about the technology as well as the basic principles. If we can be of service, please contact me privately.
Thank you for accepting my question. I'm seeking information on past and current ways digital images are gained on transmission electron microscope (TEM) e.g. digitizers, image plates, slow scan ccd It would be especially helpful to find or be directed to this information that would include logistics of image collecting, hardware, and advantages/disadvantages of each method. One recommendation as a resource was to search in the MSA proceedings journal starting around 1990. I would greatly appreciate any direct information or suggested resources. Thank you.
We have no problems with our LEO S440 software v3.04L on exporting TIF Files. I would suggest that a close look at the export setup parameters to see if something should be turned off such as compression. Our TIFs are stored as gray level no compression overlay on (for micron markers and labeling)
No one has reported any problems with image analysis software to included using extrenal scan control with our Oxford EDX system. No problems importing TIF files into CorelDraw 7, Corel Photo-Paint7 or various version of WordPerfect.
Good Luck Keith Collins
DOE Albany Research Center 1450 Queen Ave SW Albany, Oregon 97321
Dear Friends I am Young Gyu Rho who work at Chemolee Lab, a small cosmetic company, in Irving TX. I am looking for a used SEM with EDS system. If anyone has one for sale or know someone who want to sale, please let me know. Thank you.
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That is correct. Many, especially older microscopes use non-square pixels that correspond to the aspect ratio of the viewing screen. To read these images, the software needs to know about the pixel aspect ratio. If this aspect ratio is not stored in a tag of the tif file, or perhaps stored in a private tag that is not publicly known, the software cannot correctly read the image.
You may be able to adjust the pixel aspect ration in your software. Another possibility is to resample the data file with a higher frequency in one direction (use a B-spline for interpolation).
Our software, analySIS, and I am sure many of the competing products also, comes with import filters for a number of microscopes, among them Philips and LEO. This provides a transparent method for opening the files.
In some cases the manufacturers not only have non-square pixels, they store the image in several resolutions in the same tif file, and they have their own private tags for SEM or image parameters. In these cases you definitely need a dedicated import filter.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS fax: (303) 234-9271 email: info-at-soft-imaging.com
} } } ---------- } From: Lynn Rathbun[SMTP:Rathbun-at-cnf.cornell.edu] } Sent: Friday, October 2, 1998 7:48 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Questions about TIFF export under Philips XL30/TMP } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The answer to this is that the pixels in the XL30 images are not square.=20 Philips tell you that the pixels are square to within 10% (not very square= =20 in MY opinion, on our XL the images look 13% out of square if viewed in=20 Photoshop!). The upshot of this is that the TIFF images are saved at two=20 different resolutions in the x and y directions. The result is that=20 programs like NIH-Image, and Adobe Photoshop and many other Mac and PC=20 image viewing and maipulation programs cannot display the images correctly.= =20 I have found a very nice image format converter program for the Mac by a=20 programmer in Germany called Thorsten Lemke. This program is called=20 Graphic Converter and it recognizes the TIFF tags that specify the=20 different resolutions in the two directions. x=3D75dpi and 7=3D68dpi. Graph= ic=20 Converter lets you change the resolutions to match and save the file in a=20 format that NIH-Image and Photoshop etc. will understand. In fact, if you=20 pay Thorsten his shareware fee, the program will do batch conversions of=20 whole directories. Well worth the $35 shareware fee in my opinion. His=20 program can be downloaded from a variety of sites, his home site is=20 http://www.lemkesoft.de.
Of course, why an upscale program like Photoshop and a well written program= =20 like NIH-Image will not recognize these formatting tags is up for=20 discussion. Is it because Philips are using the tags the wrong way or is=20 it because the programmers of the other programs didnt ever believe that=20 someone would have non square pixels these days? I leave that for=20 discussion.
At 1:44 PM -0400 10/2/98, Scott D. Davilla wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
John Mansfield PhD CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Eventhough I tried most of your great suggestions, each and everyone of you guys are so special for helping me out; give yourselves a big hand out there in microscope cyber-space...I mean that! I will thin the sample up and stick the sample in a pool of carbon {this may affect the taste though} . Replication and vacuum perfusion will be my last resort, I'm not an expert at perfusion and I want to cheap-out on chemicals until I exhaust the other techniques.
Thanks again,
John Grazul Rutgers University Electron Imaging Facility
Referring to John Mansfield's posting on this subject: If the XL30 produces non-square pixels simply because of calibration errors (+/- 10% seems huge for a calibration error!) then perhaps the software is in fact setting the TIFF tags for x- and y-pixels per inch to be the same. (Are there such tags?). If you know the calibration error, then it is in fact possible to use Photoshop to re-sample the image to make the pixels square. Under the "Image" menu, go to "Image Size", then make sure the "Constrain Proportions" checkbox is cleared and the "Resample" box checked. Change the horizontal (or vertical - your choice) size (it doesn';t matter whether you are measuring in inches, centimeters or pixels) to make the aspect ratio correct, then click "OK". It is crude, and obviously open to introduction of artifacts during the resampling, but does give you a correctly proportioned image.
THE COURSE YOU WANT IS THE ONE RUN AT LEHIGH UNIVERSITY IN JUNE NEXT YEAR. SUGGEST YOU CONTACT MS SHARON COE AT E-MAIL ADDRESS slc6-at-lehigh.edu for details.
} From one of the course teachers
Patrick EchlinOn Fri, 2 Oct 1998, Mary Priestley wrote: Multi-Imaging Centre School of Biological Sciences University of Cambridge UK
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am in search of an introductory short course in SEM. Do you know of any } that are being offered? } } Thank you, } } Mary Priestley } Dept. of Biology } The Univ. of the South } 735 University Avenue } Sewanee, TN 37383 } } } }
A postdoctoral research position is available to study the chemistry of reactions occurring at the interface of synthetic materials with biological substrates, e.g. enamel, dentin, and bone. Applicants must be experienced in IR and/or Raman spectroscopic characterization of polymeric materials. Ph.D. in chemistry or materials science required. Please send curriculum vitae, description of research experience and three letters of reference to: Paulette Spencer DDS, PhD, University of Missouri-Kansas City School of Dentistry, 650 E. 25th St., Kansas City, MO 64108. E-mail: spencerp-at-umkc.edu.
I would be interested checking some TIFF image exports from other users. You can e-mail the TIFF images as attachments to me (not the listserver please). Please indicate some details about the image (microscope, etc.) in the e-mail. If you cannot handle the e-mail attachment on your end. I can provide a link to our ftp site where you can transfer the images. I will post a summary of results after I have a chance to check them out.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
We have here on our JEOL 8600 electron probe a NORAN EDS detector (Si(Li)) which is about 10 years of age. This spectrometer has been quite reliable, but since about a week now, I have noticed major shifts in peak energy which for a specific characteristic x-ray can be in the order of up 1 keV. The shift seem to be intermittent, and for example, after one hour, the system could come back to normal for an hour or so, and so on.
We are trying to troubleshoot this problem, and I was wondering if some would have any suggestions of where the problem may likely lie (detector, PHA, etc...??).
Thank you
Yves Thibault Research Scientist Dept. of Earth Sciences University of Western Ontario London, Ontario CANADA e-mail : ythibaul-at-julian.uwo.ca
We have been dealing with the nonsquare pixels from our XL30 for a coup= le of years now. We use Photoshop Actions to resample by using the Image Siz= e command and the following settings.
Photoshop can be set to for individual or batch modes reducing the whol= e process one single mouse click. At one time Philips had an import comm= and for MS Word that imported images and corrected the aspect ratio. Bob Anderhalt (now with Edax) may have the instructions to make this work.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-3537 USA phone: 847-938-5024 email: joe.neilly-at-abbott.com =
We have a Link eXL spectrometer and have experienced the same problem you are describing. We are still not sure what the source of the problem is, but these are some options to try for correcting the problem: 1. Frequently run a gain calibration. We use copper. 2. Condition the spectrometer to clear away contaminants. 3. Re-load the software. Good luck! ---------- } From: Yves Thibault To: Microscopy; Jackie Terry; Wayne England -----------------------------------------------------------------------.
Hi to all
We have here on our JEOL 8600 electron probe a NORAN EDS detector (Si(Li)) which is about 10 years of age. This spectrometer has been quite reliable, but since about a week now, I have noticed major shifts in peak energy which for a specific characteristic x-ray can be in the order of up 1 keV. The shift seem to be intermittent, and for example, after one hour, the system could come back to normal for an hour or so, and so on.
We are trying to troubleshoot this problem, and I was wondering if some would have any suggestions of where the problem may likely lie (detector, PHA, etc...??).
Thank you
Yves Thibault Research Scientist Dept. of Earth Sciences University of Western Ontario London, Ontario CANADA e-mail : ythibaul-at-julian.uwo.ca
We are having difficulty fixing schistomatium parasites for TEM. Standard glut/Os protocols results in shrinkage and curling of the adult worms. Does anyone have a better approach?
TIA
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
This is interesting and I didn't know about this problem. Now that I am in the market for a new SEM, I have a question to pose, Why are we as microscopists and consumers putting up with this type of crap? Is it time to press the microscope manufacturers to have a digital image format for interchange as was done for the X-ray spectra, i.e. MSA/MAS format, a number of years ago? I agree with John Mansfield, it is a no-brainer that they should be giving us square pixels and outputs with correct aspect ratios.
This question may not be appropriate for this site, but I thought it would be the quickest we to reach a large number of people. We are trying to locate a lab in the Chicago area (around the airport) with an SEM or EMP. What we hope is to have the instrument available for same day analysis of normal-to-surface samples(EDS). We are working with Alliance Steel and hoped to find a lab fairly close. If anyone can help with this matter, we'd greatly appreciate it. The dates we are looking at would be Oct 13-15th. Please respond via e-mail or call me at 513-727-5850 Armco, Inc. Thanks!!
Thanks to those that helped me with the silicon spicules in the sponge sample. Just using a diamond knife (an old one) made good sections, but soaking the block in dilute formic acid made the sections cut with less damage and I saw no change in the tissue. THANKS
I know from experience that there is no absolutes with immunolabeling with EM but my questions are "in general" 1. Has anyone tried doing immunogold labeling (post embedding) after doing DAB and peroxidase staining (before or after processing) What effect would hydrogen peroxide have on antigenic sites? We will be using UNICRYL as the embedding media polymerised by UV at 4 C. I don't have additinal tissue to experiment with. 2. When doing Double immunogold what pit fallls have people found? There are some old references (70s - 80s) that seem to be referenced with no new adaptations, Bendayan, Roth etc. I have read about using two sided and one sided. Pros & cons? Thanks for any help!
B&L do not seem to make long working distance objectives anymore. But they may be on the used market. I have been unable to locate any.
The semiconductor company that I work for, is looking for some of these for our B&L Micro Zoom scope. One lead that was a dead end at least gave me some possible catalog numbers.
B&L # 311384 25X 311385 50X 311386 50X High Res 311390 50X Ultra LWD
Any leads would be great! TIA
Phil
FYI The use for these LWD is for looking at the die inside a package while probing various signal lines. -- ********************************************************************** Philip Datner datner-at-netcom.com San Jose, CA datner-at-engmail.ulinear.com **********************************************************************
I would like to thank all responded to my call, especially Mr. ZHANG Tiejun who was very kind and helpfull, and people from JEOL US. Thank you very much! Andrew
One possible area to look at is a flucuating power supply. Does the peak always shift in the same direction, or does it flucuate around the correct eV. If it shifts in one direction then it points to a different problem.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Jackie Terry {jterry-at-ortech.on.ca} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
University of Oxford on 22-25 March, 1999 Abstract deadline: 4 December 1998
Organized on behalf of EMAG, Institute of Physics by: Prof Tony Cullis (a.g.cullis-at-sheffield.ac.uk) Dr Richard Beanland (richard.beanland-at-gecm.com =20 Co-sponsored by the Royal Microscopical Society and Endorsed by the M= aterials Research Society
**********************************
CONFERENCE SCIENTIFIC SESSIONS These will focus on the most recent advances in the application of tr= ansmission and scanning electron microscopy, SPM and X-ray diffraction to the st= udy of the structural and electronic properties of semiconducting materials.
Main topic areas: - Characterisation of as-grown semiconductors. - Investigation of lattice defect and impurity behaviour. - Study of the effects of semiconductor processing treatments. - Assessment of finished electronic devices.
Special conference sessions: - Developments in high resolution imaging and analytical transmissi= on electron microscopy. - Local area thin specimen preparation by FIB and related methods. - The nature of epitaxial layers, including quantum well, wire and = dot structures - strain relaxation, defect introduction, morphological distort= ion, self-organization, luminescence. - Wide bandgap semiconductors, especially III-V nitrides. - The structure and properties of dislocations and defect boundarie= s. - Metal-semiconductor contacts and silicides. - The effects of processing treatments. - The exploitation of advanced scanning techniques - SEM-EBIC, SEM-CL, etc - STM, AFM, SCM, BEEM, etc.
*******************
INVITED SPEAKERS H. Bender (IMEC) "Focused Ion Beam Sample Preparation" J.C. Bravman (Stanford University) "In situ Electromigration Studies" H. Cerva (Siemens) "TEM for Device Process Development" R.F. Davis (NCSU) "Growth of GaN Films" P.F. Fewster (Philips) "X-ray Diffraction Methods" E.A. Fitzgerald (MIT) "Dislocations in Graded Layers" Y Homma (NTT) "In situ SEM of Epitaxy" C J Humphreys (Cambridge University) "Advances in HREM" R. Kleiman (Lucent Technologies) "SCM of MOSFETs" J.A. Mardinly (Intel) "TEM of Devices" P. Ruternan (ISMRA, Caen) "Atomic Structure of Defects in GaN" B. Sieber (Lille University) "CL and EBIC of Heterostructures" P. Werner (MPI) "Quantum Dot Growth Phenomena"
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CONFERENCE PAPERS Contributed papers are requested in all areas indicated above. Submi= tted papers will be scheduled for either oral or poster presentation. Oral= papers will be given in a single sequence of sessions: there will be no para= llel presentations. Poster papers will be available for viewing near to t= he conference lecture theatre, with assigned times for defence by author= s.
ABSTRACT SUBMISSION The abstract deadline is 4 December 1998 and online abstract submissi= on is now available (see below).=20
CONFERENCE PROCEEDINGS The Proceedings of the conference will be published in the Institute = of Physics Conference Series, as for previous meetings. Final paper manuscripts= will be scheduled for delivery at the conference.
TRADE EXHIBITION A trade exhibition covering all semiconductor microscopy technologies= will be held on 23-24 March in areas adjacent to the conference lecture theat= re.=20 Commercial enquiries should be directed to Jacqui Watts (see below).
ADDITIONAL CONFERENCE ACTIVITIES These include: - the Annual Materials Lecture of the Royal Microscopical Society (= topic to be confirmed) - special symposium on =91Thin Sample Preparation=92 covering topic= s such as focussed ion beam methods, tripod polishing and direct cleavage techn= iques.
CONFERENCE INFORMATION Further details, including abstract submission information, can be ob= tained =66rom the conference Website http://www.iop.org/Confs. Alternatively, please contact Ms Jacquie Watts, Conferences Departmen= t, The Institute of Physics, 76-78 Portland Place, London W1N 3DH, UK. Tel: +44-(0)1865-248768; Fax: +44-(0)1865-791237; E-mail: conferences-at-io= p.org =20
This problem sounds similar to a problem I had with an old car. The engine ran well until it warmed up. It would then cough and stall. It would start again after the engine had cooled. Once the engine was hot, the car would stall again. The root cause was a loose connection in an oxygen sensor. When the engine was cold, there was sufficient contact between the wires. When hot, the contact was intermittent due to expansion and separation of the metals in the sensor connector.
Since your data acquisition hardware is ten years old, some of the components may have degraded slightly. First, take a look at the boards and look for obvious signs of damage such as fried resistors, blackened/bubbled plastic, and similar features. If possible, see if you can locate a hot spot when this problem occurs. You might be able to smell something unusual as well. If you have an infrared camera, use it. Then, I'd call Noran for help. The service engineer in my area excels at hardware diagnostics and she says the rest of the service group is better than she is.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com http:///www.sylvania.com
} Hi to all } } We have here on our JEOL 8600 electron probe a NORAN EDS detector } (Si(Li)) which is about 10 years of age. This spectrometer has been } quite reliable, but since about a week now, I have noticed major shifts } in peak energy which for a specific characteristic x-ray can be in the } order of up 1 keV. The shift seem to be intermittent, and for example, } after one hour, the system could come back to normal for an hour or so, } and so on. } } We are trying to troubleshoot this problem, and I was wondering if some } would have any suggestions of where the problem may likely lie } (detector, PHA, etc...??). } } Thank you } } Yves Thibault } Research Scientist } Dept. of Earth Sciences } University of Western Ontario } London, Ontario } CANADA } e-mail : ythibaul-at-julian.uwo.ca } }
I am a grad student at WPI. I am currently attempting to write a paper = on the above subject. Do you have any resources or leads on microbial = induced corrosion. Thanks for your assistance.
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.2106.6"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I am a grad student at WPI. I = am currently=20 attempting to write a paper on the above subject. Do you have any=20 resources or leads on microbial induced corrosion. Thanks for your =
If the shift is linear across the collected spectrum (same offset at high energy as low) I would suspect the Analog to Digital Converter reference voltage to be unstable.
If the shift is non linear, the ADC gain stability would be suspect.
I would think such a change could also result from gain changes in the detector FET/preamp.
Trouble shooting details would require a rather lengthly reply.
Good luck!
Woody White McDermott Technology, Inc. mtiresearch.com
Me: http://www.geocities.com/capecanaveral/3722
he Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi to all
We have here on our JEOL 8600 electron probe a NORAN EDS detector (Si(Li)) which is about 10 years of age. This spectrometer has been quite reliable, but since about a week now, I have noticed major shifts in peak energy which for a specific characteristic x-ray can be in the order of up 1 keV. The shift seem to be intermittent, and for example, after one hour, the system could come back to normal for an hour or so, and so on.
We are trying to troubleshoot this problem, and I was wondering if some would have any suggestions of where the problem may likely lie (detector, PHA, etc...??).
Thank you
Yves Thibault Research Scientist Dept. of Earth Sciences University of Western Ontario London, Ontario CANADA e-mail : ythibaul-at-julian.uwo.ca
Bob: I worked with those critters some years ago. They are challenging and I do not believe that you would find any "beautiful" pictures of schistomatium in the literature. I concluded that the glutaraldehyde, which is the larger molecule, does not enter through the cuticle. GA penetrates other specimens for a greater distance then does Os, but very dense cuticles may not be penetrated by GA at all. I would fix this schistosomes using Os at perhaps 30 degrees C, for half an hour, in the hope to achieve any fixation at all. The alternate approach and I never had opportunity to pursue this, would be freeze substitution or a freeze-vitrification method. I think the latter would be the most promising means to achieve better preservation. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Wednesday, 7 October 1998 1:54, "wise-at-vaxa.cis.uwosh.edu"-at-sparc5.microscopy.com [SMTP:"wise-at-vaxa.cis.uwosh.edu"-at-sparc5.microscopy.com] wrote: } } } We are having difficulty fixing schistomatium parasites } for TEM. Standard } glut/Os protocols results in shrinkage and curling of the } adult worms. } Does anyone have a better approach? } } TIA } } Bob } } } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } www.uwosh.edu/departments/biology/wise/wise.html } }
Ricky, Yes, it is possible to combine histochemical and immunocytochemical procedures successfully on the same specimen. I first stained PMN's (leucocytes) for myeloperoxidase and osmicated (preembedding) as per published protocols. The fixation was specific for immunolabelling . The cells were then embedding in LR White and sections were immunostained routinely.
Wallace Ambrose Electron Microscopy Center Dental Research Center University of North Carolina Chapel Hill, NC
I am interested in any information or experience that you might have or had with the printers above for reproduction of photo quality prints. Cost per page (for B&W), print integrity, etc. I am looking to purchase a printer to defray the cost of film for more routine analysis.
Any information is appreciated.
David Rose
======================== David BG Rose W.L. Gore and Associates Elkton, MD
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This posting is a response to a communication between Hildy Crowley and me concerning which is preferable for dehydration...acetone or ethyl alcohol. It originated as a question I had regarding her procedure for membrane fixation which was a response to a thread asking about use of maleate buffers. Other comments/opinions would be welcomed. ==========
In response to whether to use acetone or alcohol as preferred dehydrant, there does not seem to be a definitive answer. Some of the older texts seem to lean on the side of acetone because it appears to extract less phospholipid. However, depending on the type of tissue you are fixing, that may not be the only, or even the most important type of intracellular lipid. Ethanol appears to extract less hydrophobic lipids than acetone. The type of fixation is critical for preservation of lipids (as well as proteins and other cell components) for each specific tissue. Time of dehydration also appears to be important to extraction rates.
For those that want to review the literature, a good place to start is some of the general reference books (Bozzola & Russell-Electron Microscopy(1992); Dykstra-Biological Electron Microscopy(1992); Hayat-Fixation for Electron Microscopy(1981); Hayat-Principles and Techniques of EM(1981); etc.).
I have always tried different fixative-buffer combinations with new tissues before deciding on a specific protocol since what works for one sample is not always optimum for another. Perhaps we should take the little extra time to check dehydration agents as well. I prefer using ethanol since it is less volatile, less toxic and does not take up water quite as quickly as acetone....however, I also am open to using whatever will give superior final results.
Debby ===================== Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
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FALL MEETING OF THE METROPOLITAN MICROSCOPY SOCIETY
Time: 9:30 am (registration begins)
Place: Radisson Inn, 601 From Rd., Paramus, NJ, (201) 262-6900
Directions: Garden State Parkway to Exit 165 (E. Ridgewood Ave./Oradell Ave.) From Road is on the west side of the Garden State Parkway, and adjacent to it.
AGENDA =======================
* 9:30 - 10:30 Registration -- $20 (includes buffet lunch). PLEASE PRE-REGISTER SO WE CAN GET A PROPER ESTIMATE FOR THE BUFFET LUNCH! (See contacts at bottom of notice)
*************************************** * Coffee and Danish sponsored by JEOL * ***************************************
* 10:30 - 10:45 Introductory Remarks and society business (Phil Flaitz).
* 10:45 - 11:30 GOING NONDISPERSIVE, Kurt Heinrich, National Institute of Standards and Technology, Gaithersburg, MD.
* 11:30 - 12:15 THE APPROACHING REVOLUTION IN X-RAY MICROANALYSIS: ENERGY DISPERSIVE SPECTROMETRY WITH SILICON DRIFT DETECTORS AND MICROCALORIMETERS, Dale E. Newbury, National Institute of Standards and Technology, Gaithersburg, MD
* 1:00 - 1:45 EDS FROM THEN TILL NOW---A CHRONOLOGY OF INNOVATION, John Friel, Princeton Gamma-Tech, Princeton, NJ.
* 1:45 - 2:30 SOME PRACTICAL CONSIDERATIONS IN THE ANALYSIS OF EDS SPECTRA AT LOW ENERGIES AND LOW VACUUM, Bob Anderhalt, EDAX Inc., Mahwah, NJ.
* 2:30 - 3:15 PROGRESS TOWARDS ATOMIC-RESOLUTION X-RAY MICROANALYSIS, David Williams, Department of Materials Science and Engineering, Lehigh University, Bethlehem, PA.
For more information OR TO PRE-REGISTER please contact either:
Phil Flaitz or Evan Slow (914) 892-3094 (201) 760-2524 flaitz-at-us.ibm.com ess-at-feico.com
Philip L. Flaitz IBM Analytical Services Ph.......(914) 892-3094, FAX -2003 flaitz-at-us.ibm.com
by mail.unixg.ubc.ca with smtp (Exim 1.92 #1) id 0zQw6a-0000I3-00; Wed, 7 Oct 1998 09:06:24 -0700 Message-Id: {2.2.32.19981007160636.008e9f60-at-pop.interchange.ubc.ca} X-Sender: mager-at-pop.interchange.ubc.ca X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Dear Yves, I had a similar problem with a Kevex that turned out to be a broken wire inside the high-voltage bias supply. The wire to the connector was broken but hidden under the shrink-wrap insulation. If it touches momentarily you will get good calibration, but after a while with no bias your peaks will droop. Hope this helps. You wrote: } Hi to all } } We have here on our JEOL 8600 electron probe a NORAN EDS detector } (Si(Li)) which is about 10 years of age. This spectrometer has been } quite reliable, but since about a week now, I have noticed major shifts } in peak energy which for a specific characteristic x-ray can be in the } order of up 1 keV. The shift seem to be intermittent, and for example, } after one hour, the system could come back to normal for an hour or so, } and so on. } } We are trying to troubleshoot this problem, and I was wondering if some } would have any suggestions of where the problem may likely lie } (detector, PHA, etc...??). } } Thank you } } Yves Thibault
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
It was asked whether etching LR White would help solve high background problems. LR White is an acrylic. Acrylics are far less crosslinked and far more hydrophilic than epoxies. Moreover, osmium is generally not used with this system. Etching LR White could be done, but it almost never is. It probably would not withstand the process very well - even epoxies develop holes and thin out. Your first goal in immunoAu is to get signal. Got signal? Great! Increasing the concentration of antibody is likely to cause more background than wanted. Need to lower background? Dilute the antibody stepwise for starters. Get any good immuno text and follow the suggestions for lowering background one by one. Not enough signal then? Try increasing signal stepwise as instructed in immuno texts. Etching the LR will not solve your background problems. It is likely to be a big mess. Bye, Hildy
I am looking for a motorized drawer (like a CD rom Drawer) that is used for microscope slides. Does anyone know of such a unit or where I might be able to look. Any help or info would be greatly appreciated.
A good start: ASM Metals Handbook Volume 13: Corrosion. The whole ASM Metals Handbook series is probably in the WPI library.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com http:///www.sylvania.com
} -----Original Message----- } From: John Rhatigan [SMTP:jrhatigan-at-entwistleco.com] } Sent: Wednesday, October 07, 1998 7:37 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Microbial Induced Corrosion } =20 } I am a grad student at WPI.=A0 I am currently attempting to write a = paper on } the above subject.=A0 Do you have any resources or leads on microbial } induced corrosion.=A0 Thanks for your assistance.
The references I sent out for the maleate buffer systems required acetone for dehydration. I have used the system for llamelar bodies in lung tissue. These structures have a high phospholipid component. We tried the acetone, we tried the alcohol. Acetone was slightly better than alcohol for this purpose. But the most astonishing and unexpected preservation improvement appeared when the tissue was fixed overnight in the refrigerator in a fresh quantity of osmium. That effect was truly startling. Bye, Hildy
John, One area that comes to mind involves the biofilm corrosion of ocean = going boats' copper paint bottoms. I have seen some posters on biofilms = attacking copper at a MSA meeting a few years ago and it also involves work the = marine paint industry is interested in as well. Good luck, Dave Audette audette-at-osi.sylvania.com
} -----Original Message----- } From: John Rhatigan [SMTP:jrhatigan-at-entwistleco.com] } Sent: Wednesday, October 07, 1998 7:37 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Microbial Induced Corrosion } =20 } I am a grad student at WPI.=A0 I am currently attempting to write a = paper on } the above subject.=A0 Do you have any resources or leads on microbial } induced corrosion.=A0 Thanks for your assistance.
Sounds like a problem for Click and Clack ---------- } From: Crossman, Harold To: Microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
We have both. HP LaserJet's which we use for viewing B&W images. The 600 dpi output is more than good enough for most work and we seldom go to film anymore. You won't have any trouble getting someone who has one to print a BMP or other photo out to show you how good they look. I also have an Epson 1440x720 dpi color printer for near photo quality in color, and I seldom use it for that, although I just bought a digital camera. I also have a 300 dpi HP 1600color inkjet which I use the heck out of because its fast and good enough for initial work like cropping and framing and positioning. Film is fast becoming sidelined in all the branches of our company, and I see the same thing wherever I go. I am getting a 1200 dpi LaserJet with my next machine so my B&Ws will look even better. The metallurgists are jealous because they are stuck with an expensive, slow photo printing system which uses proprietary file formats. They will be upgraded someday.
I don't know anything about Alps. HP is so easy to set up (network too) and has the fastest drivers, Epson had to outdo them big in dpi to get my attention.
Tim Chavez Boeing Chem Characterization Lab
____________________________________________________ It is those people who have been good to you that you should try to get even with.
} ---------- } From: } "drose-at-wlgore.com"-at-Sparc5.Microscopy.Com[SMTP:"drose-at-wlgore.com"-at-Sparc } 5.Microscopy.Com] } Sent: Wednesday, October 07, 1998 8:27 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Printers - Epson vs HP vs Alps? } } } I am interested in any information or experience that you might have } or had } with the printers above for reproduction of photo quality prints. } Cost per } page (for B&W), print integrity, etc. I am looking to purchase a } printer } to defray the cost of film for more routine analysis. } } Any information is appreciated. } } David Rose } } } ======================== } David BG Rose } W.L. Gore and Associates } Elkton, MD } } } } } }
I wish to find out when to use digital deconvulation software vs confocal real time vs confocal scanning microscopy. The application is to visualize the intracellualr location of FITC tagged oligonuceotides obtained from animals treated with it in vivo. The second application is to characetrize the movement of cells in response to chemotactic agents. These are very slow movements, but the cells have to be live.
What is a good general criterion for evaluation of a core microscopy facility in an academic situation? What kind of ranking priority may be assigned to different operations, such as research and service, management of the core facility, cost recovery (user fee), teaching commitment (regular and short courses), publications, etc.
A user fee is levied in most labs for use of major instruments. The sample preparation methods may influence the amount of time a particular instrument is in use, specially a TEM. What would be an average estimate, in terms of percent time spent per year on TEMs, SEMs and Confocals in a core facility?
Randy Mandryk Microtechnique Lab, Room CW 225T EXT 3473, Biological Sciences University of Alberta
} In response to whether to use acetone or alcohol as preferred } dehydrant, there does not seem to be a definitive answer ..........
Quite right, there does not seem to be a simple or definitive answer to this. Years ago we did some work on the extraction of C14 labelled components from cells fixed and dehydrated with different protocols. The final conclusion was that there is not much difference in general extractive power of acetone versus ethanol, but what came out clearly was that the 70% step (acetone or ethanol) is the great extractor. This step extracts 10x as much as the anhydrous step. Leaving the material in 70% ethanol or acetone does serious damage to its structure and composition. The ref is: Coetzee & van der Merwe, 1989: Extraction of Carbon 14-labeled compounds from plant tissue during processing for electron microscopy. J Electron Microsc. Technique, 11:155-160.
Jan C
Prof Jan Coetzee Head: Lab for Microscopy and Microanalysis Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-362-5150 Pretoria 0002, South Africa http://www.up.ac.za/science/electron/emunit1.htm
As a SEM user and responsible for lab equipment, I'm looking for = scientific and technical informations about what's new and which are the = last subjects discussed all around our world. I'm only registered to one = journal "microscopy and analysis" (because it is free of charges, of = course...) I would like to know which are the most common journals you read to get = knowledge in new SEM techniques, applications (I'm especially interested = in VPSEM) and also in X-ray analysis.What are the interest/concern of = each journal, publication delay for papers after being submitted and so = on...
As I don't need to receive each whole journal (and I can't afford the = whole fees neither) and that University is for me too costly in time, my = question is : Is there any way to get the abstracts from any web site? or at least the = current contains? May I register somewhere to receive last submitted = papers abstracts or at least titles with the e-mail?
Thank you for answers. Jean-Francois COULON,=20 Materials Department, ecole d'ingenieurs Louis de Broglie Campus de Ker Lann 35170 Bruz, France. direct e-mail : materiaux-at-ecole-debroglie.fr
Another paper which contains valuable information on the extraction of various radiolabeled components (from bacteria) during substitution is: L.L.Graham, T.J.Beveridge (1990) J.Bacteriol. 172, 2141-2149 Evaluation of freeze-substitution and conventional embedding protocols for routine electron microscopic processing of eubacteria.
regards, Reinhard Rachel
Dr. Reinhard Rachel Universitaet Regensburg Lehrstuhl fuer Mikrobiologie (Prof. Dr. K.O. Stetter) D - 93040 Regensburg Tel.: xx49-941-943-4534 Fax.: xx49-941-943-1824 http://www.biologie.uni-regensburg.de/Mikrobio/Stetter/Gruppen/rachel.html e-mail: Reinhard.Rachel-at-biologie.uni-regensburg.de
Crowley's statement about the importance of fixation, in my experience, is correct. Previous work (20 years ago) on multilamellar bodies in lung indicated a slight difference in choice of dehydrants. Ethanol and acetone alone seemed to extract lipids similarly. However, a mixture of ethanol and acetone (70:30) worked better. Further, dehydration was done at 4 degrees C with tissue cut to {1mm cubes, 2 minutes per step and put into 100% Spurr before warming to room temperature. Beware of condensation ruining the Spurr. I might choose LR White or Gold now.
Hello ,microscopists does anybody know e-mail and the name of the chairman of "Uni-Export Instruments Ltd" (UK)? Please, mail to gusev-at-ipm.sci-nnov.ru
Best regards, Sergey mailto:gusev-at-ipm.sci-nnov.ru
First I want to thank all that sent TIFF images exported from their microscopes. I also want to thank Dirk van der Wal of Philips in Eindhoven for a very quick reply to my specific questions about the XL30 and TIFF export.
Now the results. Yes, on some microscopes, the TIFF exports are images with non-square pixels. However, all the images that have non-square pixels also have x and y resolution tags that if used will present the image in the correct aspect ratio. For example, our XL30 (in standard definition mode) exports a 712 x 484 pixels image that is really 4:3 in visual aspect ratio. The x-resolution tag is 89/3 or 29.667 pixels/cm and the y-resolution tag is 26.889 pixels/cm. If the image processing program applies this scaling, then the image will now look correctly. If it does not (and many do not), then the image looks distorted. Why is this done. Well it has to do with how (and when) the framestore in the microscope was designed. Back in the non-digital days, square pixels on the viewing crt and photo crt was the important issue. Even though the actual scan might not be with square pixels, it could be corrected later in analog display. Surprisingly this also tends to be true with the newer digital microscopes. Back to the one I really know, the XL30, why 712 x 484. Why not a true 4:3 for the framestore. I suspect that the reason has to do with NTSC/PAL video output. The XL has a natural NTSC (for N.America) video output. Rather than scan convert the framestore to NTSC/PAL, the framestore uses clever tricks to output either NTSC or PAL video. The side effect is that if you save the digital contents of the framestore and display it assuming square pixels, the image looks distorted.
Is this a misuse of the TIFF tags. Well not really. According to TIFF 6.0 spec, the use of these tags is optional and can be applied to display, printing or both. Since there is both an x and y tag, this implies that the values can be different. Unfortunatly, about 99% of the image processing programs (both Mac and Win) either ignore the tags or assume that their values are the same (square pixels). It's kind of like pixels can have TIFF legal floating point values but how many image processing programs know what to do with a floating point pixel? Not very many. Are non-square pixels bad? Yes and no. If you do not know that they are non-square, that is very bad. If you know and can tolerated rescaling, then they are not a problem. Just rescale and go. The real problem is the 1) not many imaging programs understand non-square pixels and 2) sometimes this issue is presented as a calibration problem.
So far, only Graphic Converter (thanks John) at http://www.lemkesoft.de. is the only program that I have found that correctly understands and uses these tags. It can also be used to rescale to square pixels. You can also rescale manually in other image processing programs. Philips supplies (thanks Dirk) a Win program called XLstretch that will also correct the TIFF image to square pixels. I know the next time I write a TIFF reader, I too will understand and apply these tags.
Now with the mechanics behind, what of the other issues. Philips exports the raw contents of the framestore without correcting the aspect ratio. This allows reloading into the framestore without rescaling. Well I have to agree with them on this point. Images are data to me and I don't like rescaling very much. Other microscope makers export rescaled images and don't let you know that it was rescaled. In some applications, this whole issue is not important as long as there is some way to rescale the image for display/printing. In others (like mine), the data is the most important part. I cannot rescale or I will contaminate the measurements I'm trying to make. What I will do now is adjust my other equipment to match the pixel aspect ratio. That way I can do data to data comparisions.
So the real result is if TIFF export is important for your use and you require original data then you must ask these two questions; 1) Are the contents of the TIFF file rescaled in anyway? 2) Does the TIFF image have non-square pixels.
If the answer is yes, then the TIFF export is not going to help you and you really need some type of external active scan device that will acquire the type of images you need.
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Madelaine Rodriguez wrote: ================================================== Can anyone direct to a source for Feon FT. I need it to clean an ultrathin window on my EDS system. =================================================== This product was deemed to be a big time ozone eater! That is why you can't find it anywhere, it is illegal to sell it, at least in most countries.
The "replacement" (so far as I know the only one) is called Asahiklin AK 225 . It is a product of Asahi Chemical in Japan. The price however will be a shock to you. But you can purchase it from a firm called Tech Spray, Inc., PO Box 949, Amarillo, TX 79105 Ph: (806) 372-8523. Just make sure you are sitting down when you hear the price. You will wish for the days (and the pricing) of TF!
With regard to its safety in terms of cleaning the window of your EDS system, that I can give no guarantees.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The references I sent out for the maleate buffer systems required acetone for dehydration. I have used the system for llamelar bodies in lung tissue. These structures have a high phospholipid component. We tried the acetone, we tried the alcohol. Acetone was slightly better than alcohol for this purpose. But the most astonishing and unexpected preservation improvement appeared when the tissue was fixed overnight in the refrigerator in a fresh quantity of osmium. That effect was truly startling. Bye, Hildy
Only real photo quality is dye sublimation (kodak, codonics, tektronics, etc) which are expensive (maybe $5K and up). Inkjets like you name, with special, expensive glossy paper and 1200 dpi are almost as good for a few hundred $. I like the Epson Stylus Photo (EX) myself. All will be color (but make good B&W) unless you get a laser printer. Dave Pevear, Houston
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am interested in any information or experience that you might have or had } with the printers above for reproduction of photo quality prints. Cost per } page (for B&W), print integrity, etc. I am looking to purchase a printer } to defray the cost of film for more routine analysis. } } Any information is appreciated. } } David Rose } } ======================== } David BG Rose } W.L. Gore and Associates } Elkton, MD
Does anyone have current info on what are the requirements for publishing digital micrographs? Has ther ben developed an industrial standard or does each journal have it's own requirements. Are the standards different for TEM vs. SEM? We are looking into digitlzing our microscopes and it is important that this modification is done appropriately so that we obtain a system that does more than produce images for the internet.
Bill
William R.McManus Supervisor Electron Microscope Facility Department of Biology Utah State University Logan UT 84322-5305
My latest opinion- We went 'round and 'round and evaluated all kinds of printers for not just the EM Facility, but our entire research group. We were not deliriously happy with any of them, and finally selected an Epson Stylus Photo (ca. $300) and a Tektronix something dye-sub for ca. $7,000. The Epson is slightly better (!), but the prints are not waterproof.
Here's the punchline: right after finally committing our monies, I found printers that I *far* prefer above all the rest - the Fujix Pictrography 3000 and 4000. They are not dye-sublimation, but use a silver halide technology, making them more like actual photographs than the others. They should be archivable, and the only chemical they require is distilled water. They are cost competitive, and the output is totally awesome! These printers are the best I've seen. The 3000 has come down in price to {$10K. I'd buy one in a heartbeat if I had the money.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Be very careful using AK-225 for cleaning windows. It is a stronger solvent than Freon, especially for polymers. I'm not sure what polymer the windows are made with, but AK-225 has some definite incompatibilities. AK-225 also has a relatively low exposure limit of 50 ppm.
There are several "Freon" or CFC-113 replacements on the market. None are exactly like CFC-113, but the ones closest to it are AK-225 (hydrochlorofluorocarbon), 3M's HFE 7100 (hydrofluoroether) and DuPont's Vertrel XF (hydrofluorocarbon). Another replacement are the perfluorocarbons from 3M (like PF5052), but I don't believe the general public can get these any more. As Chuck indicated, all are expensive. In drum quantities they are $175-250/gallon. They are also all available as various mixtures and azeotropes to increase their solvency power, since most are not very strong solvents.
I've spent the last 8 years (and a LOT of $) working with replacements for CFC-113 for precision cleaning, so I'm pretty familiar with what's available.
I would not use AK-225 without compatibility studies or the manufacturer's okay. I'd also be hesitant to recommend anything without compatibility testing.
You may want to check some chemical supply houses as you may be able to still get small quantities of CFC-113 for laboratory use.
John Giles Principal Materials Engineer Honeywell Space Systems
This product was deemed to be a big time ozone eater! That is why you can't find it anywhere, it is illegal to sell it, at least in most countries. The "replacement" (so far as I know the only one) is called Asahiklin AK 225 . It is a product of Asahi Chemical in Japan. The price however will be a shock to you. But you can purchase it from a firm called Tech Spray, Inc., PO Box 949, Amarillo, TX 79105 Ph: (806) 372-8523. Just make sure you are sitting down when you hear the price. You will wish for the days (and the pricing) of TF! With regard to its safety in terms of cleaning the window of your EDS system, that I can give no guarantees. Chuck
} Can anyone direct to a source for Feon FT. I need it to clean an } ultrathin window on my EDS system. } Madelaine Rodriguez } AlliedSignal
I will soon use up the last of the Freon which I use for this purpose, and I intend to try a hydrocarbon, perhaps hexane. Recent correspondance on this list does indicate that the oil buildup is predominantly rotary pump oil, so a hydrocarbon solvent should work pretty well without, I hope, being too aggressive to the window-mounting cement. Any detector manufacturers got anything to add on this?
cheers
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I've seen posts which responded to the original "HP vs Epson vs Alps" which imply Alps is an ink jet. The original poster may have been referring to the Alps 1300 which is a dual mode 600dpi ink jet and dye-sub printer for ~$500. I was personally impressed with the example Alps sent me, but I also hear it may be the slowest dye-sub on the market ... and we all know printer performance has more to do with how well the printer interfaces with the OS. I know nothing beyond this ... but certainly am curious if someone has practical experience.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Since MOXTEK makes most of the thin windows used I should step forward here and say what I know, but I defer to the x-ray detector manufacturers, since only they know which windows were put into which detectors.
We have done some cleaning experiments here, which is appropriate since only we can afford to clean a window till it breaks. We have not tried every solvent, however, but if it is important to you let us knowknow and we will try it.
We like freon too. The only alternate solvents that we have tested are methanol and acetone. These don't damage the window as long as the following rules are met:
1. Use pure solvents so you don't leave a residue 2. Don't let the solvent sit or puddle on the window 3. Don't squirt the window directly
Our method is to point the "snout" of the detector down, at an angle of about 30 degrees. Then we squirt the solvent at the mount (not the window), letting it run across the window by gravity. If you don't let the acetone sit on the epoxy for very long it does not hurt it.
Best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
I have two customers that are looking for older equipment. Specifically a Cambridge S-150 SEM and a Philips 300 TEM or equivalent. Please e-mail me if you have any leads. The SEM does not need to be in working order. The customer wants a "parts" machine for his existing S-150.
Dear Madelaine, If you are referring to Freon FT, this is difficult to obtain, since it is now banned from use by all signers of the Montreal Accord to reduce the use of ozone-destroying materials. The USA and Canada are both signers. You should ask your EDX manufacturer, but my thin-window snout can be cleaned with iso-propyl alcohol, then carefully air-dried.
You wrote: } Can anyone direct to a source for Feon FT. I need it to clean an } ultrathin window on my EDS system. } Madelaine Rodriguez } AlliedSignal } } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
I have followed the discussion about the best printer. I have a slightly different take. I have three color ink jets of different makes available to me. My question is what is the best "photographic paper" to use in an ink jet. I have tried Polaroid, Kodak, and HP. I have opinions but only qualitative. Has someone done a comprehensive study about photographic inkjet papers?
Thanks for considering this request.
Blystone in Texas
-------------------------------- Robert V. Blystone, Ph.D. rblyston-at-trinity.edu
Department of Biology Trinity University 715 Stadium Drive San Antonio, Texas 78212 210.736-7243 FAX 210/736-7229
Unfortunately competing publishers are not likely to agree=20 to join forces and publish, even journal contents by=20 subject on the internet. There are, however, abstracts or=20 contents listings available for various journals by=20 particular publishers. If you go to the links in our online=20 (append /links to our address) and use find for=20 "journal" you will get three URLs, including Elsevier's=20 content listing for hundreds of journals. I suppose if you=20 find Springer's equivalent the majority of EM journals=20 would be covered. Cheers Jim Darley ProSciTech Microscopy=20 PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au=20 *****
On Thursday, 8 October 1998 20:04, Jean-Fran=E7ois COULON=20 [SMTP:materiaux-at-ecole-debroglie.fr] wrote: } } Hi to all } } As a SEM user and responsible for lab equipment, I'm } looking for scientific and technical informations about } what's new and which are the last subjects discussed all } around our world. I'm only registered to one journal } "microscopy and analysis" (because it is free of charges, } of course...) } I would like to know which are the most common journals } you read to get knowledge in new SEM techniques, } applications (I'm especially interested in VPSEM) and } also in X-ray analysis.What are the interest/concern of } each journal, publication delay for papers after being } submitted and so on... } } } As I don't need to receive each whole journal (and I=20 can't } afford the whole fees neither) and that University is for } me too costly in time, my question is : } Is there any way to get the abstracts from any web site? } or at least the current contains? May I register } somewhere to receive last submitted papers abstracts or } at least titles with the e-mail? } } Thank you for answers. } Jean-Francois COULON, } Materials Department, } ecole d'ingenieurs Louis de Broglie } Campus de Ker Lann } 35170 Bruz, France. } direct e-mail : materiaux-at-ecole-debroglie.fr }
The references I sent out for the maleate buffer systems required acetone for dehydration. I have used the system for llamelar bodies in lung tissue. These structures have a high phospholipid component. We tried the acetone, we tried the alcohol. Acetone was slightly better than alcohol for this purpose. But the most astonishing and unexpected preservation improvement appeared when the tissue was fixed overnight in the refrigerator in a fresh quantity of osmium. That effect was truly startling. Bye, Hildy
I find that many operators and field service engineers use IPA (isopropyl alcohol) for EDS window cleaning. Some clean as often as every two weeks! It doesn't attack window glues like acetone might. They use the flow down and over technique. If they are careful it works. If they are too nervous about the process or break too many windows then they call me or check out our web site for a product that cleans differently.
More information: http://www.msa.microscopy.com/SM/XEI/XEIHomePage.html
Ronald Vane XEI Scientific 3124 Wessex Way Redwood City, CA 94061 650-369-0133
Robert, Your different take is valid. I have tried a dozen kinds, HP, Epson, Canon, Different Mill specialties. I have two answers for you. 1. 90 % of what I print on goes onto the plain old office stock because I am just evaluating the report for fit, crop, etc, the details that print preview always fails to clue me in on. Or else the image is instructive enough in the lower resolution mode that I can print on that paper mostly. 2. When I need to approach photo quality, I find I need to match the manufacturer's recommendation, even though the paper for higher than 720 dpi is $0.50 per sheet. This is pretty glossy paper, but the 720 paper doesn't really show the printer weaknesses in 1440 mode since the ink runs into the paper's imperfections which are similar in size to the small dots. I can get some practice scraps from our in house photo-xerox lab, which help me to demonstrate the benefits so my boss will let me order it. Still I seldom use it since our photo's need to be of instructive quality to start with. I do run into problems of properly printing images of slightly mottled panels, but I find that I am just as limited by my source file as I am by my printer's talents. And I have to relate this to you. The clearest smoothest image I ever printed, was on some free labels that came with my Korpak bottles. 12 labels to a sheet, I still have that beautiful image today. But here's something else you have to consider. I used to have a Canon 360 dpi printer and didn't think much of it. Just before I got rid of it, a friend emailed me a large photo. Because the source had so many bits in it, the canon stunned us as it had never had so much detail in the file to turn into an image before. I think it was a 10,000 by 10,000 pixel image from a fancy scanner [exageration]. We stared at the printed photo in disbelief for weeks wondering how we had misjudged the printer so badly, or how to duplicate such photo quality when we wanted it. I believe _____ Microscopy files are seldom even as big as 640x480. That's our challenge, make a good printable picture from that, just don't blow it up too big. Papers seldom limit us, and most customers are satified with low res paper unless you are doing dollar bill intaglio. [ididntsaythat]
I am continually trying new stock and have had GREAT luck with the shiny squares of white cardboard that I take out of my tshirt and underwear packages. Please send me yours (the cardboard not your underwear). These papers have the serious advantage of being very absorbent so the images dry quickly. When I tried printing on a cello window of an envelope, my Epson ink wouldn't dry for hours. So the dots puddled and gave me an artsy useless image that smeared too easily. Be careful, your printer needs a thick paper setting to handle card stock or it may be expensively damaged. I tore some shiny blank pages out of some books tossed by the local library [software engineering texts, 2 years old and uselessly outdated] and got great images on them. The yellow edges made a great image for my family photos. Many ads in the mail are glossy white and have a blank side and I seem to have a graffiti artist's disdain for empty spaces. So I feed it to my printer and finally commit that ______ photo to paper. I know there are other's out there who live by the goof rules and are always stretching these boundaries. Lets here from you guys. I'll bet there are some ladies who have tried printing on vacuum cleaner bags and cereal boxes. Where are you guys who have printed on their six pack cardboard and oil filter packages?
I am thinking about peroxide etching some trash bags and trying them ;-)
Tim Chavez His goofship tonight. (up too late, ... but you asked !!!)
[most of this reply is serious Bob]
} ---------- } From: Robert Blystone[SMTP:rblyston-at-trinity.edu] } Sent: Thursday, October 08, 1998 5:53 PM } To: Microscopy Listserver } Subject: Best printers, best paper } } ---------------------------------------------------------------------- } -- } To the List: } } I have followed the discussion about the best printer. I have a } slightly } different take. I have three color ink jets of different makes } available } to me. My question is what is the best "photographic paper" to use in } an } ink jet. I have tried Polaroid, Kodak, and HP. I have opinions but } only } qualitative. Has someone done a comprehensive study about } photographic } inkjet papers? } } Thanks for considering this request. } } Blystone in Texas } } -------------------------------- } Robert V. Blystone, Ph.D. } rblyston-at-trinity.edu } } Department of Biology } Trinity University } 715 Stadium Drive } San Antonio, Texas 78212 } 210.736-7243 FAX 210/736-7229 } }
Hi all Can people tell me their favourite method for adjusting thickness of formvar films. I'm trying concentration variation but without much success. If concentration variation is the method of choice what are suitable % ranges to try. Many thanks
Chris
Chris Gilpin Experimental Officer Biological Sciences EM Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 0161 275 5170 Fax +44 0161 275 5171 Chris Gilpin http://130.88.91.187/emunit
Michael referred to an Alps 1300, with which we have no experience, however the Alps MD2300 is both a dye-sub and dry transfer heat printer with a 600 dpi resolution (not an inkjet). They refer to the latter process as microdry. It sold for about $700, but may not be their current model.
The dye-sub is quite good, requiring their special paper and cartridges, but extremely slow, a full page photograph taking about 3/4 hour at this resolution. Lower resolution is faster, as is the microdry process which can be used on papers of differing quality. We have had trouble in feeding extremely rough paper through the machine. At 600 dpi resolution, the four color microdry prints take about 5 min.
I don't sell these machines! Len
} I've seen posts which responded to the original "HP vs Epson vs Alps" } which imply Alps is an ink jet. The original poster may have been } referring to the Alps 1300 which is a dual mode 600dpi ink jet and } dye-sub printer for ~$500. I was personally impressed with the example } Alps sent me, but I also hear it may be the slowest dye-sub on the } market ... and we all know printer performance has more to do with how } well the printer interfaces with the OS. I know nothing beyond this ... } but certainly am curious if someone has practical experience. } } cheerios, shAf } } {} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} } Michael Shaffer, R.A. - ICQ 210524 } Geological Science's Electron Probe Facility - University of Oregon } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/ } } } } } Leonard Zablow Howard Hughes Medical Institute 722 West 168 St. New York, N.Y. 10032
I have a ceramic sample that I want to do some carbon EDX analysis on. I have always used methanol or ethanol to clean my samples. My question is will my carbon EDX analysis be affected by the use of any of these organic solvents as a cleaning agent; if so, how much. Is there a better cleaning agent I should use to give me a more accurate result for carbon analysis.
The Windows 95 accessory program 'Imaging For Windows by WANG' can correctly read the x and y resolution tags in TIFF files. I use it to quickly view image files from our rectangular pixel Zeiss DSM982 FESEM.
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
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RE} publication of digital images 10/9/98 8:56 AM Bill,
You pose a very good question. We have been going through a number of tri= als with our publisher on this very topic. We also have been scanning 35= mm slides into digital format for journal publication.
After a number of iterations we have solved at least some of the problem.= Our publisher needs 300 dpi files so the final published picture(s) don'= t look pixelated.=20
I am sure that someone out there has had some experience with this whole = question, especially John Russ. His Image Processing book was done comple= tely digitally.
The resolution and depth of the files that are captured digitally are muc= h higher than our publisher needs them. The trick is to give the publishe= r just enough data so that their printers can print a good image. It does= n't make any sense to give them an extremely high resolution image if the= y only need 300 dpi/lpi.
I am very interested in hearing from others.
John Shane
--------------------------------------
Dear List;
Does anyone have current info on what are the requirements for publishing=
digital micrographs? Has ther ben developed an industrial standard or do= es each journal have it's own requirements. Are the standards different fo= r TEM vs. SEM? We are looking into digitlzing our microscopes and it is important that this modification is done appropriately so that we obtain = a system that does more than produce images for the internet.
Bill
William R.McManus Supervisor Electron Microscope Facility Department of Biology Utah State University Logan UT 84322-5305
I have a ceramic sample that I want to do some carbon EDX analysis on. I have always used methanol or ethanol to clean my samples. My question is will my carbon EDX analysis be affected by the use of any of these organic solvents as a cleaning agent; if so, how much. Is there a better cleaning agent I should use to give me a more accurate result for carbon analysis.
Hi microsopists, I am having a problem with Spurr resin. The blocks are coming out very brittle. I am careful when I weigh out the components and polymerize at 65C. What else would cause brittle blocks? Another problem is that staining with 1% T blue with 1% borax is very poor. Any suggestions would be of help. Thanks. -- Regards, Gregory Rudomen Technical Specialist University Microscopy Imaging Center State University of New York at Stony Brook 516-444-3126 Greg-at-umic.sunysb.edu ********************************************************** Standard disclaimer: The opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. **********************************************************
I would like to hear more about permanence of images made by different technologies. I am using a wax transfer printer (Sony Mavigraph), which produces fairly attractive glossy pictures on proprietary paper. Paper + "ribbon" cost is, I believe } $1 per print. The material smells unpleasant, but more important, it transfers to adjacent surfaces. Although the original images that have transferred don't seem to lose much in quality, I am concerned about permanence and am interleaving glassine (not easily found in the right size, half of a Euro letter size) to reduce this.
FWIW, I recently saw a demo of a small, relatively cheap dye sub printer, bundled with a camera, that supposedly has better permanence. The picture quality appeared acceptable for record rather than presentation purposes.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
Try Microtome Service Co 7568 Florian Way Liverpool, NY 13088 (315) 451-1404
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Matthew thomas cimino To: Microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------.
I am trying to locate a source for Sorvall MT-1 Microtome parts. Please let me know if any are available.
Hi to all, I have just changed the solutions in our developer and fixer baths that serve our two TEMs. Unfortunately, the first couple of users have informed me that their negatives are next to useless and that the negatives are pink around the edges! I'm pretty confident that the developer is ok but the fixer put at my disposal (Agfa Structurfix) was not what I used last time (Ilford Hypam). Any ideas? Many thanks, Andy
__________________________________ Dr Andy Burrows Materials Science and Engineering Department of Engineering The University of Liverpool Liverpool England L69 3BX
Instead of a Freon, you might consider using a low-boiling grade of petroleum ether, or a pure hydrocarbon such as heptane, for cleaning detector windows. It appears from all I have been able to determine that the oil contaminant on EDS detectors is usually hydrocarbon fragments from the oil-sealed rotary mechanical backing pump, and as such would be readily soluble in a hydrocarbon solvent, while the epoxy cements should be very little affected by it. Acetone and the various alcohols, on the other hand, will attack epoxies to some extent, although probably not significantly in the short time they would be exposed in the cleaning process usually used. On the other hand, acetone and the alcohols are not particularly good solvents for hydrocarbons. The old rule form chem lab is, "Like dissolves like".
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Usually brittleness in Spurr's is caused by moisture in the resin. For example, I once allowed some specimen vials to remain open overnight in a rotator - to enhance penetration. Bad idea. Result: brittle blocks that shattered during trimming. Even traces of moisture in Spurr's will cause this. In ordinary epoxies, this is not a problem (Epon for example could be left open overnight or several days without this problem developing).
} I am having a problem with Spurr resin. The } blocks are coming out very brittle. I am careful } when I weigh out the components and polymerize at } 65C. What else would cause brittle blocks? } Another problem is that staining with 1% T blue } with 1% borax is very poor. Any suggestions would } be of help. Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
At 02:13 PM 10/9/98 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Chris,
The way we do this is by taking a graduated cylinder with a constricted end that fits a glass slide (to conserve solution), filling it to the proper depth with the formvar solution, and then capping the cylinder to let the atmosphere inside become saturated with the evaporating solvent. The slide is dipped into the formvar, then lifted out and left dangling in the saturated atmosphere for varying amounts of time. This allows the formvar to drain off the glass surface without drying out. The longer the drain time, the thinner the film.
A paper clamp on a piece of wire is all you need to hold the slide, and the wire allows the tube to remain mostly capped as you pull the slide up out of the solution. Crude, but it works.
Let me know if you have any questions. Hope it helps.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Chris Gilpin asked the following: ==================================================== Can people tell me their favourite method for adjusting thickness of formvar films. I'm trying concentration variation but without much success. If concentration variation is the method of choice what are suitable % ranges to try. ==================================================== I consulted with the chief of our quality assurance in our filmed grid production area. There was some disagreement between telling all our "secrets" vs. not telling all our secrets. But here are the "secrets" which perhaps others might find of value. But keep in mind that specific technique probably has a lot to do with it as well and then there is that intangible called "art". So I can recite the "recipe" but not the "art" that has to be learned from practice:
For most filmed grid production, we use 0.25 to 0.3 % Formvar(TM) in ethylene dichloride. What is not generally appreciated is that there are a number of different grades of the resin called Formvar, but so far as we know, only one of them is optimum for the filmed grid application. If one was using other grades, they might require different concentrations for the best results. The grade that we use is the grade that we sell for this purpose. And no, we won't tell that little secret.
For thicker films, we would increase that concentration to the range 0.4 to 0.5%. Thicker films are usually made when one wants us to make a holely film with bigger holes. We find that a concentration above about 0.7% results in a film that is literally too thick for any EM application.
If we want thinner films, then one reduces the concentration. But we are already making the films at the borderline in minimum thickness anyhow (because that is what our customers demand) but the concentration could be reduced a bit more to about 0.2%. However, below 0.2%, the film becomes just too succeptible to cracking, so that becomes the lower limit for reducing the thickness.
But anyone who has made filmed grids will attest, the real test, e.g. where the tire hits the pavement, so to speak, is how the Formvar film behaves in the vacuum of the TEM and under exposure to the electron beam. These percentages and other aspects of the technique are all optimized in order to produce optimum performance, e.g. long lasting and virtually no drift. But one really does need a TEM right there, next to where the films are being made, so that they can be instantly inspected and corrective action taken, right there on the spot, e.g. by way of a concentration change. If this is not done, the the QC step end up being when you gear up to do your experiment and then you find out the grids are not stable. Variables such as temperature, humidity and perhaps other factors seem to come into play, so that the optimum concentration one day might not quite be the optimum concentration on some other day. There is even one school of thought that believes that light slowly degrades the solid Formvar which typically might be quite old, since it is used so sparingly.
Disclaimer: SPI Supplies has produced custom coated filmed grids for TEM for customers worldwide for some number of years. More information about the coating of TEM grids can be found on our website given below.
Chuck
================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Extend the fixative time. You could try to refix the pink-edged negatives. They may clear but not if it has been too long in the light. Best of luck,
Chuck --- On Fri, 9 Oct 1998 17:32:32 +0100 Andy Burrows {ab0895-at-LIVERPOOL.AC.UK} wrote: -------------------------------------------------------------- ---------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi to all, I have just changed the solutions in our developer and fixer baths that serve our two TEMs. Unfortunately, the first couple of users have informed me that their negatives are next to useless and that the negatives are pink around the edges! I'm pretty confident that the developer is ok but the fixer put at my disposal (Agfa Structurfix) was not what I used last time (Ilford Hypam). Any ideas? Many thanks, Andy
__________________________________ Dr Andy Burrows Materials Science and Engineering Department of Engineering The University of Liverpool Liverpool England L69 3BX
-----------------End of Original Message-----------------
------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
Hi Chris,
I just copy the method from the lab manual which I wrote for my TEM course as follows:
-clean the slide with clean lens paper. -dip slide into formvar (0.3 % w/v solution in chloroform). -draw back the slide slowly (the faster the motion, the thicker the film) and dry the slide vertically (about 1 min). -cut the membrane parallel to the edge of the slide with a razor blade. -dip the slide ( with scored side up ) into water slowly at a 30 degrees angle and let the slide sink down to the bottom of the deep dish. The film should float off onto the water. -place grids (polished side up) on the floating film. -a piece of paper or parafillm is dropped onto the film. -pick up the paper and leave it in a dust-free Petri dish.
The thickness of film can be judged by the light reflection as the method for thin sections. Grey or silver-greay film are good for coating.
Good luck,
Ming
On Fri, 9 Oct 1998, Chris Gilpin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all } Can people tell me their favourite method for adjusting thickness of formvar } films. I'm trying concentration variation but without much success. If } concentration variation is the method of choice what are suitable % ranges } to try. } Many thanks } } Chris } } } } Chris Gilpin } Experimental Officer } Biological Sciences EM Unit } G452 Stopford Building } Oxford Road } Manchester } M13 9PT } phone +44 0161 275 5170 } Fax +44 0161 275 5171 Chris Gilpin } http://130.88.91.187/emunit } } }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * #1074B Dentistry Pharmacy Building * * University Of Alberta. * * Edmonton, Alberta, Canada T6G 2N8 * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Here's a summary of the responses I received from my TEM inquiry. THANKS TO ALL OF YOU WHO HELPED ME!!
There were few responses, and it appears to me the data are mainly anectotal. I think the two people who may have authoritative data (or best available) are Barbara Foster and Elinor Solit. Both earn their livings as consultants, and since my interest isn't critical enough to fund a "real" study, I didn't feel justified in asking them for further efforts at this time. I think most of us know them already, but for the record: Elinor Solit: {cambrex%world.std.COM} Barbara Foster: {mme%map.COM} I have no stake in either's services.
The responses: NIH -at- Bethesda, MD ~300 (in 1987) UC Berkeley 10 TEM's active Hawaii 5 Canada ~200 China ~1000 United States ~1000 JEOL units World ~4000 to 5000, 1/3 USA, 1/3 Europe, 1/3 Asia
Chris Gilpin wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Hi all } Can people tell me their favourite method for adjusting thickness of formvar } films. I'm trying concentration variation but without much success. If } concentration variation is the method of choice what are suitable % ranges } to try. } Many thanks } } Chris } } Chris Gilpin } Experimental Officer } Biological Sciences EM Unit } G452 Stopford Building } Oxford Road } Manchester } M13 9PT } phone +44 0161 275 5170 } Fax +44 0161 275 5171 Chris Gilpin } http://130.88.91.187/emunit
-- For Ladd substrates we do the following:
1. Dipping method: 0.25 - 1%
2. Drop on water method: 1 - 2.5%
3. Increase thickness in method 1 by a multiple dipping process.
Charles Duvic, Chemist LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
Seneca College is now accepting bids for used EM related equipment from the Bio. Chem department. Please click on the URL provided to view the items available. Thank you.
{http://www/biochemsale/jeol.html} {http://www/biochemsale/jeol.html} Clickto view larger image Seneca College of Applied Arts & Technology has a number of used pieces of electron microscopy laboratory equipment for sale. The most notable of which is a {http://www/biochemsale/jeol.html} JEOL100CX Transmission Electron Microscope. (Click photo to view larger image) Bids for this equipment are being solicited under the following Terms and Conditions. Seneca College's Asset Disposal Policy is in effect throughout the bidding process. All sales are final. All items sold "As-Is, Where-Is" with no warranty given or implied. All offers are subject to sales taxes, where applicable. Payment to Seneca College, and removal of goods from premises within three (3) working days of acceptance of offer. The successful bidder must remove goods from existing positions using a properly insured heavy-equipment mover, if necessary. Seneca College does not assume any responsibility for damage or injury resulting from the use of items purchased. Seneca College reserves the right to accept or reject any, or all bids.
Inspection Date: Monday, October 19, 1998, beginning 11:00am On site contact: Laurel Schollen 416-491-5050 ext 2390 (e-mail; Laurel.Schollen-at-senecac.on.ca) Bids must be received prior to 3:00pm (local time) October 30, 1998 If you require additional information about this Equipment Sale, please contact: Chris Legros, Buyer Seneca College, 416-491-5050 ext 7141 (e-mail; {mailto:Chris.Legros-at-senecac.on.ca} Chris.Legros-at-senecac.on.ca), or John Shannon, Senior Buyer, Seneca College, 416-491-5050 ext 7144 (e-mail; {mailto:John.Shannon-at-senecac.on.ca} John.Shannon-at-senecac.on.ca)
Bids for the BioChem Equipment Sale can be submitted one of two ways:1) Print out and complete the form below, send by facsimile: 905-479-1281, Attention: Chris Legros 2) Fill out the electronic form below and submit via e-mail by selecting the "Submit" button at the end of the form. Within the form below, complete the "Bid Price" for as many items as required. Please complete the contact information at the end of this form. Electron MicroscopesItem #Bid Price Transmission Electron Microscope JEOL #100CX, Serial #Em156134-05 ( {http://www/biochemsale/jeol.html} seephoto)1A Scanning Electron Microscope JEOL #35C SEM, Serial #Em150017-78FK (for parts only)1B Transmission Electron Microscopy Ancillary EquipmentItem #Bid Price LKB knife breakers 1, Serial #7801B2A LKB knife breakers 2, Serial #7801B2B LKB knife breakers 3, Serial #7801B2C Riechert OMU3 microtome2D Riechert OMU3 cryo-cut unit, Serial #151-FC22E Microtome LKB Productor 12F Microtome LKB Productor 22G Microtome 2, Sorval MT-1(1)2H Microtome 3, Sorval MT-1(1)2I Microtome 4, Sorval MT-1(1)2J Microtome 5, Sorval MT-1(1)2K Fresh Tissue Cutter, Serial #V9108902L Vacuum Embedding Oven 1 (rotary vacuum pumps attached), Serial # Thelco 20-Z-72M Vacuum Embedding Oven 2 (rotary vacuum pumps attached), Serial # Thelco model 192N Riechert Embedding System 1, Serial # KT-1002O Riechert Embedding System 2, Serial # KT-1002P Evaporator 1, Baltzers BA32Q Evaporator 2, Baltzers BA3 Serial #14812R Automatic slide dipper (for EM)2S LKB pyramatome2T Sorval JB-4 2U Film Dessicator (rotary vacuum pumps attached)2V LKB Huxley ultramicrotome2W Photographic Darkroom SuppliesItem #Bid Price Durst Point light source enlarger, Serial #138-S3A Photographic Enlarger (Omega)3B Photo Mounting Press, Serial #J175863C Print drier for resin coated paper, Iford, Serial #1050-RC3D (4x) grain focusing aids3E Scanning Electron MicroscopyItem #Bid Price TriVac Rotary Vacuum Pump, D&A, Serial #12913086824A Critical Point Drying apparatus4B Sputter Coater-Hummer V4C Name: Title: Address: City: Prov/State: Telephone:Area codeTelephone:Extension: After completing the above, click "Submit" to send your bid, or "Reset" to clear all fields in the form.
There seems to be all kinds of problems with digital deconvulation software, most notably the time needed to obtain good images. I was interested in finding out if anyone had experience with a digital deconvulation software that was rapid. I have heard that the one from Improvision is rapid, but has someone used it and found it to be good?
I would not change Formvar concentration outside the range of 0.2-0.3% (w/w). (I use only ethylene dichloride; it seems to give better separation of film from the slide). The thickness of film depends on the speed of withdrawal of the slide from the solution; the faster, the thicker film. To make thin film, withdraw the slide from the solution by SLOW and STEADY motion; but NOT out of the jar yet. Drain excess Formvar solution by holding the slide in the vapor above the solution for 15 seconds. This will help you getting more uniform thickness of the film.
Good Luck!
Seung-Geuk Shin SUNY-ESF at Syracuse sgshin-at-syr.edu
============================== } -----Original Message----- } From: Chris Gilpin [mailto:cgilpin-at-fs1.sem.man.ac.uk] } Sent: Friday, October 09, 1998 9:14 AM } To: microscopy-at-sparc5.microscopy.com } Subject: formvar film thickness } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all } Can people tell me their favourite method for adjusting thickness } of formvar } films. I'm trying concentration variation but without much success. If } concentration variation is the method of choice what are suitable % ranges } to try. } Many thanks } } Chris } } } } Chris Gilpin } Experimental Officer } Biological Sciences EM Unit } G452 Stopford Building } Oxford Road } Manchester } M13 9PT } phone +44 0161 275 5170 } Fax +44 0161 275 5171 Chris Gilpin } http://130.88.91.187/emunit } } }
I had the same problem, when I used an old batch (3-5 years with occasional uses), or one exposed to moisture (If you keep the bottles in a low temperature for extended shelf life, let it warm before opening the bottle). With bottles out of the box, I had no problem. In my opinion, Spurr's has relatively poor cutting properties, even when the block itself is not brittle. I usually successfully mix Spurr's (complete homogeneous mixture including DMAE) with Epox 812 (complete homogeneous mixture including DMP-30) to improve cutting properties, with minimal sacrifice in Spurr's excellent viscosity. It's been very cooperative for plant tissues.
Seung-Geuk Shin SUNY-ESF at Syracuse sgshin-at-syr.edu
============================================ Disclaimer: The opinions expressed in this communication are solely my own. ============================================
} -----Original Message----- } From: Greg R [mailto:greg-at-umic.sunysb.edu] } Sent: Friday, October 09, 1998 11:11 AM } To: microscopy } Subject: Spurr resin } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi microsopists, } I am having a problem with Spurr resin. The } blocks are coming out very brittle. I am careful } when I weigh out the components and polymerize at } 65C. What else would cause brittle blocks? } Another problem is that staining with 1% T blue } with 1% borax is very poor. Any suggestions would } be of help. Thanks. } -- } Regards, } Gregory Rudomen } Technical Specialist } University Microscopy Imaging Center } State University of New York at Stony Brook } 516-444-3126 } Greg-at-umic.sunysb.edu } ********************************************************** } Standard disclaimer: The opinions expressed in } this } communication are my own and do not necessarily } reflect those of the University Microscopy Imaging } Center. } ********************************************************** } }
I would not change Formvar concentration outside the range of 0.2-0.3% (w/w). (I use only ethylene dichloride; it seems to give better separation of film from the slide). The thickness of film depends on the speed of withdrawal of the slide from the solution; the faster, the thicker film. To make thin film, withdraw the slide from the solution by SLOW and STEADY motion; but NOT out of the jar yet. Drain excess Formvar solution by holding the slide in the vapor above the solution for 15 seconds. This will help you getting more uniform thickness of the film.
Good Luck!
Seung-Geuk Shin SUNY-ESF at Syracuse sgshin-at-syr.edu
} -----Original Message----- } From: Chris Gilpin [mailto:cgilpin-at-fs1.sem.man.ac.uk] } Sent: Friday, October 09, 1998 9:14 AM } To: microscopy-at-sparc5.microscopy.com } Subject: formvar film thickness } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all } Can people tell me their favourite method for adjusting thickness } of formvar } films. I'm trying concentration variation but without much success. If } concentration variation is the method of choice what are suitable % ranges } to try. } Many thanks } } Chris } } } } Chris Gilpin } Experimental Officer } Biological Sciences EM Unit } G452 Stopford Building } Oxford Road } Manchester } M13 9PT } phone +44 0161 275 5170 } Fax +44 0161 275 5171 Chris Gilpin } http://130.88.91.187/emunit } } }
Randy I hope you get some good responses. I am interested in some of the same questions. Talking to two labs in the USA I seem to be giving away my services, but I need examples of cost recovery, number of personnel, samples per year, costs for current services to compare and substantiate any cuts in services or increases in costs. Another issue is, our chairman now wants lists of publications that (only) include EM micrographs (this is to indicate the value of the core labs). How many labs have to keep that information? I don't believe a core facility should be expected to see that an experiment gets published. I feel lucky if they remember to acknowledge the lab. Plus, work done as abstracts, thesis, presentations, and posters are also important. Am I alone here?
My faculty lab director and I are looking for labs around the US, especially the mid west that would be willing to discuss this type of data and send info through the mail for security if need be. We need to do this fairly soon to get cost increases in place for upcoming grant deadlines. Thanks
Rick Vaughn M.S.
Electronmicroscopy Research Facility Dept. Cell Biology & Anatomy Univ Neb Med Ctr (402) 559-7347 RLVAUGHN-at-MAIL.UNMC.EDU
I have the same problem with JB-4 Plus resin blocks. They are very brittle and tend to shatter during trimming which is very irritating. They have been left in a vaccum dessicator in vials without lids on. The blocks have set with small bubbles on the underside, a problem I haven't previously had using JB-4, not sure why?
On Saturday, 10 October 1998 6:28, Garber, Charles A. [SMTP:cgarber-at-priam.chesco.net] wrote: . . . .There was some disagreement between telling all our "secrets" vs. not telling all our secrets. But here are the "secrets" which perhaps others might find of value. . . . The message is clear: Chuck & Co know how to make filmed grids better than anybody else. Sadly, I learned nothing from the "secrets", I wonder in fact: what were the secrets?
. . . . What is not generally appreciated is that there are a number of different grades of the resin called Formvar, but so far as we know, only one of them is optimum for the filmed grid application. If one was using other grades, they might require different concentrations for the best results. The grade that we use is the grade that we sell for this purpose. And no, we won't tell that little secret. . . . . Oh, now I learn that I was just lucky that all of the films that I had made over about 30 years worked well, yet none of the formvar came from Chuck. But why his reply? The questioner had no problems with formvar but was interested in technique to control thickness.
. . . . .But one really does need a TEM right there, next to where the films are being made, so that they can be instantly inspected and corrective action taken, right there on the spot, e.g. by way of a concentration change. If this is not done, the the QC step end up being when you gear up to do your experiment and then you find out the grids are not stable. .. . . . Other suppliers know nothing and get their films probably made at the back of a soup kitchen. You take a grave risk if you buy your films anywhere else. But why? The question had been on how to control film thickness and not were to buy filmed grids.
. . . .Disclaimer: SPI Supplies has produced custom coated filmed grids for TEM for customers worldwide for some number of years. More information about the coating of TEM grids can be found on our website given below. . . . . This is not a disclaimer, but another vintage, bold-faced advertisement by Chuck. Nobody had asked about his filmed grids and he gave no useful information on how to control film thickness well. Two other writers provided methods ("double dipping" and "lengths of time after dipping in solvent saturated atmosphere") that he may find useful to apply to his assembly line. He is too cheap to pay for advertising his filmed grids and has again used the microscopy server instead.
Non italics by Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
Andy: The fixer was too weak (or exhausted). It is a good idea to periodically check fixer. Place a small piece of undeveloped film in the fixer, full light does not matter. The film will become quite clear in under 30 seconds if its really good. If it takes a minute it's exhausted or made-up too weak. Exhausted fixer could be used to prefix film for a couple of minutes before using good fixer. This will prolong the good fixer's life. A brief water rinse after developing and before fixing is a good idea. I don not like acetic acid because with some materials it can cause mottling. Minimum fixation time is three times the time film takes to clear, but I suggest to not fix for less than three minutes. The colouration of under fixed materials only sometimes disappears when refixing. Certainly, underfixed images should be refixed if useful parts of the neg were unaffected and they too would change if not refixed promptly. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Saturday, 10 October 1998 2:33, Andy Burrows [SMTP:ab0895-at-LIVERPOOL.AC.UK] wrote: } Hi to all, } I have just changed the solutions in our developer and } fixer baths } that serve our two TEMs. Unfortunately, the first couple } of users } have informed me that their negatives are next to useless } and that } the negatives are pink around the edges! I'm pretty } confident that } the developer is ok but the fixer put at my disposal (Agfa } Structurfix) } was not what I used last time (Ilford Hypam). Any ideas? } Many thanks, } Andy } } __________________________________ } Dr Andy Burrows } Materials Science and Engineering } Department of Engineering } The University of Liverpool } Liverpool } England } L69 3BX } } Tel: +(44) (0)151 794 5372 } Fax: +(44) (0)151 794 4675 } email: ab0895-at-liv.ac.uk
Elisabeth, Some years ago I saw a note in a journal saying: that blocks soften (for a while) and that trimming of brittle blocks is improved, if the blocks are soaked for 24 hours in abs. ethanol. I think that note referred to Spurr's but guess it would work for others. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Saturday, 10 October 1998 12:07, Cox, Elizabeth [SMTP:CoxE-at-prose.dpi.qld.gov.au] wrote: } I have the same problem with JB-4 Plus resin blocks. They } are very brittle } and tend to shatter during trimming which is very } irritating. They have } been left in a vaccum dessicator in vials without lids on. } The blocks have } set with small bubbles on the underside, a problem I } haven't previously had } using JB-4, not sure why? } } Liz } } }
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Jim., I feel your response to Charles Garber's posting was out of line.
You have compounded what may/may not have been a legitimate attempt to help with a totally useless response. You only served to emphasize your demonstrated lack of tolerance. I, for one, would appreciate you're keeping your personal opinions about other list serve members (which came through loud and clear from your sarcastic phrasing) to yourself. Sometimes silence is golden!! Nestor polices this listserve and I am sure will notify offenders of the rules of the listserve if he deems them out of line.
Chuck was right-on about checking films in a TEM. This is especially important when trying to vary thickness as either too thick or too thin can be problematic. The original question may have come from an inexperienced user who, in his/her desire to make thin/thick films, could end up with something quite useless....reminder to check results as you go is not irrelevant information.
Neither is a warning that there are different grades of formvar and what works for one may not work for others. Success in regulating thickness definitely starts with the formvar used, humidity, solvent, "art" of the user, etc....I have often seen where two people working side-by-side will use identical solutions and method yet produce very different results.
Disclaimer: I do not have any relationship with anyone at SPI and only occasionally purchase from that company.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
On Saturday, 10 October 1998 6:28, Garber, Charles A. [SMTP:cgarber-at-priam.chesco.net] wrote: . . . .There was some disagreement between telling all our "secrets" vs. not telling all our secrets. But here are the "secrets" which perhaps others might find of value. . . . The message is clear: Chuck & Co know how to make filmed grids better than anybody else. Sadly, I learned nothing from the "secrets", I wonder in fact: what were the secrets?
. . . . What is not generally appreciated is that there are a number of different grades of the resin called Formvar, but so far as we know, only one of them is optimum for the filmed grid application. If one was using other grades, they might require different concentrations for the best results. The grade that we use is the grade that we sell for this purpose. And no, we won't tell that little secret. . . . . Oh, now I learn that I was just lucky that all of the films that I had made over about 30 years worked well, yet none of the formvar came from Chuck. But why his reply? The questioner had no problems with formvar but was interested in technique to control thickness.
. . . . .But one really does need a TEM right there, next to where the films are being made, so that they can be instantly inspected and corrective action taken, right there on the spot, e.g. by way of a concentration change. If this is not done, the the QC step end up being when you gear up to do your experiment and then you find out the grids are not stable. .. . . . Other suppliers know nothing and get their films probably made at the back of a soup kitchen. You take a grave risk if you buy your films anywhere else. But why? The question had been on how to control film thickness and not were to buy filmed grids.
. . . .Disclaimer: SPI Supplies has produced custom coated filmed grids for TEM for customers worldwide for some number of years. More information about the coating of TEM grids can be found on our website given below. . . . . This is not a disclaimer, but another vintage, bold-faced advertisement by Chuck. Nobody had asked about his filmed grids and he gave no useful information on how to control film thickness well. Two other writers provided methods ("double dipping" and "lengths of time after dipping in solvent saturated atmosphere") that he may find useful to apply to his assembly line. He is too cheap to pay for advertising his filmed grids and has again used the microscopy server instead.
Non italics by Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
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-----Original Message----- } From: Chris Gilpin [mailto:cgilpin-at-fs1.sem.man.ac.uk] Sent: 10 October 1998 13:14 To: Me at home
On Saturday, 10 October 1998 6:28, Garber, Charles A. [SMTP:cgarber-at-priam.chesco.net] wrote: . . . .There was some disagreement between telling all our "secrets" vs. not telling all our secrets. But here are the "secrets" which perhaps others might find of value. . . . The message is clear: Chuck & Co know how to make filmed grids better than anybody else. Sadly, I learned nothing from the "secrets", I wonder in fact: what were the secrets?
. . . . What is not generally appreciated is that there are a number of different grades of the resin called Formvar, but so far as we know, only one of them is optimum for the filmed grid application. If one was using other grades, they might require different concentrations for the best results. The grade that we use is the grade that we sell for this purpose. And no, we won't tell that little secret. . . . . Oh, now I learn that I was just lucky that all of the films that I had made over about 30 years worked well, yet none of the formvar came from Chuck. But why his reply? The questioner had no problems with formvar but was interested in technique to control thickness.
. . . . .But one really does need a TEM right there, next to where the films are being made, so that they can be instantly inspected and corrective action taken, right there on the spot, e.g. by way of a concentration change. If this is not done, the the QC step end up being when you gear up to do your experiment and then you find out the grids are not stable. .. . . . Other suppliers know nothing and get their films probably made at the back of a soup kitchen. You take a grave risk if you buy your films anywhere else. But why? The question had been on how to control film thickness and not were to buy filmed grids.
. . . .Disclaimer: SPI Supplies has produced custom coated filmed grids for TEM for customers worldwide for some number of years. More information about the coating of TEM grids can be found on our website given below. . . . . This is not a disclaimer, but another vintage, bold-faced advertisement by Chuck. Nobody had asked about his filmed grids and he gave no useful information on how to control film thickness well. Two other writers provided methods ("double dipping" and "lengths of time after dipping in solvent saturated atmosphere") that he may find useful to apply to his assembly line. He is too cheap to pay for advertising his filmed grids and has again used the microscopy server instead.
Non italics by Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
Dear list Jim Darley has a point. I was asking about ways of controlling film thickness. Here follows the background to my request which will clarify further...
I have a "customer" in my core facility embarking on a serial sectioning project. She needs to use slot grids and has chosen the .75 variety. For whatever reason the films are not lasting as far a prolonged viewing in the microscope. Yes we are carbon coating. Yes we have tried carbon coating before and after section collection. Yes we have tried glow discharging with and without carbon. If anything our films are a bit on the thin side (great for some applications but not this one!) We have produced some thick films but they are very prone to wrinkling. Now you will see the reason for the question. I would like to make thicker films than silver/gold but without wrinkling. Perhaps I am guilty of asking the bare question without detailing why. I would add however that all replies are greatly received. Members of the list never cease to surprise me with their willingness to help. Other posts to the list show that most of us are fighting for our lives to stay in "business". As a community we stand a better chance of survival if we are all successful. Sharing knowledge is part of making our labs a success. I appreciate that commercial companies have a living to make and can be forgiven for keeping trade secrets. As long as the rest of us don't fall into that trap.
Again many thanks to ALL who replied. Long live EM
Chris
Experimental Officer Biological Sciences EM Unit University of Manchester Oxford Road Manchester M13 9PT Phone +44 (0)161 275 5170 Fax +44 (0)161 275 5171
What is available on video that contains footage of membracid sized organisms filmed in motion through an electron microscope? Is there anything available you would suggest from the MSA Video Library? Your help would be greatly appreciated! Thank you, Mark Diegel
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School: Providence College Question: I've had so many questions answered through this service over the last 2 years - It's wonderful.
I'm a senior biology undergrad looking forward to continuing my education in grad school - hopefully a Ph.D program in cell biology. Who out there has opinions on institutions with exceptional microscopy facilities? I'm really into TEM/SEM but would love to experience some more advanced techniques and equipment. I really want a program that's interested in answerering questions of cell biology by pushing the limits of microscopy and related tools. Where should I look?
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Mark -
The radiation used for imaging in an electron microscope is lethal. There's a lot of good light microscopy available on the WWW. Look at the website list in the MICRO bibliography (URL below); start with the list at "K-12 microscopy resources". "Cells Alive" might be a good place to begin.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Dear fellow microscopists: Could anyone direct me to a reliable antibody making facility here in the US? Thank you. Linda Chicoine Viatech Imaging Ivoryton, CT lchicoine-at-snet.net
Yes, and before the radition destroyed the advancing horde of the attacking nanocreatures the vacuum pump of the electron microscope rendered them lifeless - Ed Sharpe in the temple of doom
Doug - } } I'm a senior biology undergrad looking forward to } continuing my education in grad school - hopefully } a Ph.D program in cell biology. Who out there has } opinions on institutions with exceptional microscopy } facilities? I'm really into TEM/SEM but would love } to experience some more advanced techniques and } equipment. I really want a program that's interested } in answerering questions of cell biology by pushing } the limits of microscopy and related tools. } Where should I look?
I just visited the Keck Imaging Lab at Arizona State; very impressive & worth looking into (pardon the pun!).
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I apologize for providing incomplete information. I usually mix them 1 to 1 by weight. But you can reduce the amount of Epox 812 mixture, if low viscosity is important, as in freeze-dried tissues. It's been tested primarily on plant tissues, fixed chemically or lipophillized, and worked well for both. It worked on some chemically fixed animal tissues (small intestine & liver from a frog). I don't have much experience with animal tissues and I am not sure about lipophillized animal tissues. But, I can say generally Spurr's provides better infiltration, while Epox 812 offers better cutting properties (via better adherence to the tissue). There has been no negative effect of mixing two, as far as I know. Thus, I guess any tissue, which are not incompatible with Epox or Spurr's, can be safely embedded in the mixture of the two. ----------------------------------------------- I also forgot including the staining method with toluidine blue in the previous message. If it is to stain 'thick section' for preliminary observation, place a drop of your 1% toluidine onto the section, heat it on a 70 C-hot plate until you see thin dry edge around the stain, wash the stain gently with squirts of water.
For microautoradiographic purpose, I use following method. This applies to 1 um-sections affixed on a gelatin-coated slide. 1) Dip the slides in 10% paraformaldehyde for 1-2 min. 2) Rinse the slides in distilled water for 1 min (or rinse twice for 30 sec. each). 3) Stain sections by dipping the slide for 1/2 to 1 min in 0.05%-0.1% toluidine blue prepared in 0.2M Na-phosphate buffer (pH 7.0). (Stain may precipitate over the time; shake and refilter it.) 4) Wash the slide in running water quietly for about 2 min or shorter period (prolonged washing will result very faint staining). 5) Air-dry the slide in vertical position.
I hope this helps.
Regards,
Seung-Geuk Shin
} -----Original Message----- } From: Tamara Howard [mailto:howard-at-cshl.org] } Sent: Saturday, October 10, 1998 3:07 PM } To: Seung-Geuk Shin } Subject: RE: Spurr resin } } } Do you mix the two at a ratio of 1:1? Aso, do you use the mixture for } tissues from } animals, plants, or both? } } Thanks! } } Tamara Howard } CSHL } } } On Fri, 9 Oct 1998, Seung-Geuk Shin wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I had the same problem, when I used an old batch (3-5 years } with occasional } } uses), or one exposed to moisture (If you keep the bottles in a low } } temperature for extended shelf life, let it warm before opening } the bottle). } } With bottles out of the box, I had no problem. } } In my opinion, Spurr's has relatively poor cutting properties, } even when the } } block itself is not brittle. I usually successfully mix Spurr's } (complete } } homogeneous mixture including DMAE) with Epox 812 (complete homogeneous } } mixture including DMP-30) to improve cutting properties, with minimal } } sacrifice in Spurr's excellent viscosity. It's been very cooperative for } } plant tissues. } } } } Seung-Geuk Shin } } SUNY-ESF at Syracuse } } sgshin-at-syr.edu } } } } ============================================ } } Disclaimer: The opinions expressed in this } } communication are solely my own. } } ============================================ } } } } } -----Original Message----- } } } From: Greg R [mailto:greg-at-umic.sunysb.edu] } } } Sent: Friday, October 09, 1998 11:11 AM } } } To: microscopy } } } Subject: Spurr resin } } } } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } Hi microsopists, } } } I am having a problem with Spurr resin. The } } } blocks are coming out very brittle. I am careful } } } when I weigh out the components and polymerize at } } } 65C. What else would cause brittle blocks? } } } Another problem is that staining with 1% T blue } } } with 1% borax is very poor. Any suggestions would } } } be of help. Thanks. } } } -- } } } Regards, } } } Gregory Rudomen } } } Technical Specialist } } } University Microscopy Imaging Center } } } State University of New York at Stony Brook } } } 516-444-3126 } } } Greg-at-umic.sunysb.edu } } } ********************************************************** } } } Standard disclaimer: The opinions expressed in } } } this } } } communication are my own and do not necessarily } } } reflect those of the University Microscopy Imaging } } } Center. } } } ********************************************************** } } } } } } } } } } } } } }
To all, does anyone out there have the lens current values for the old Philips EM 200 EM. The Mag. is off in several steps and we nedd the currents of the IL, DIFF. & PROJ. Lenses for each step. Any help is greatly appreciated. Peter Stolzenberg, Pesto Inc. 215-699-6160, FAX 215-699-5275 pesto-at-erols.com
I apologize for providing incomplete information. I usually mix them 1 to 1 by weight. But you can reduce the amount of Epox 812 mixture, if low viscosity is important, as in freeze-dried tissues. It's been tested primarily on plant tissues, fixed chemically or lipophillized, and worked well for both. It worked on some chemically fixed animal tissues (small intestine & liver from a frog). I don't have much experience with animal tissues and I am not sure about lipophillized animal tissues. But, I can say generally Spurr's provides better infiltration, while Epox 812 offers better cutting properties (via better adherence to the tissue). There has been no negative effect of mixing two, as far as I know. Thus, I guess any tissue, which are not incompatible with Epox or Spurr's, can be safely embedded in the mixture of the two.
-----------------------------------------------
I also forgot including the staining method with Toluidine Blue in the previous message. If it is to stain 'thick section' for preliminary observation, place a drop of your 1% Toluidine Blue onto the section, heat it on a 70 C-hot plate until you see thin dry edge around the stain, wash the stain gently with squirts of water.
For microautoradiographic purpose, I use following method. This applies to 1 um-sections affixed on a gelatin-coated slide. 1) Dip the slides in 10% paraformaldehyde for 1-2 min. 2) Rinse the slides in distilled water for 1 min (or rinse twice for 30 sec. each). 3) Stain sections by dipping the slide for 1/2 to 1 min in 0.05%-0.1% Toluidine Blue prepared in 0.2M Na-phosphate buffer (pH 7.0). (Stain may precipitate over the time; shake and refilter it.) 4) Wash the slide in running water quietly for about 2 min or shorter period (prolonged washing will result very faint staining). 5) Air-dry the slide in vertical position.
I hope this helps.
Regards,
Seung-Geuk Shin
} -----Original Message----- } From: Tamara Howard [mailto:howard-at-cshl.org] } Sent: Saturday, October 10, 1998 3:07 PM } To: Seung-Geuk Shin } Subject: RE: Spurr resin } } } Do you mix the two at a ratio of 1:1? Aso, do you use the mixture for } tissues from } animals, plants, or both? } } Thanks! } } Tamara Howard } CSHL
Maybe I missed the posting, but when developing and printing one of the most important points that has not been raised is that of agitation!
Many years ago when I ran the Hitachi demo labs in the UK, I took advice = on our photography from Kodak and Ilford. It was to lift the film out of th= e developer/fixer after 15 seconds and then to tilt it to the left (15 seconds later to the right) for 2 seconds. This was done throughout developing, but a surprise to me, for the first minute and a half of fixing. They did not want a stop bath to be used but water at the same temperature as the other chemicals. We called this the dunk and tilt method.
My problem was patchy negatives though poor technique!
One other point was that when using this technique, as I pestered all our=
clients to follow, the next problem was fogged films. Why, because no o= ne ever changes their safe lights! However on the packet it always says tha= t they are only safe for a fixed period of time due to fading (~ 5 years). =
My clients all complained that they had never had this problem before usi= ng the dunk and tilt method and when questioned they almost all had 10 years=
old plus safe lights!
I hope that this is a tale worth telling?
Steve Chapman Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
I'm looking for a book(s) that describes the theoretical and practical aspects of EDX for our Unit library. The books should contain information on the use of EDX for biological applications (botanical - eg. sea grass secretions; & zoological - eg mussel tissues in pollution studies) and for the analysis of marine/lake/riverine silts/sediments. I'm sure there are numerous books on these subjects - which ones would you guys like to see on your shelves?
Thanks in advance
Mike Gregory Professor Mike Gregory Electron Microscope Unit, University of Durban-Westville Private Bag X54001 Durban, Natal, South Africa 4000
} } Hi all: } } I'm looking for a book(s) that describes the theoretical and practical aspects of EDX for our Unit library. The books should contain information on the use of EDX for biological applications (botanical - eg. sea grass secretions; & zoological - eg mussel tissues in pollution studies) and for the analysis of marine/lake/riverine silts/sediments. I'm sure there are numerous books on these subjects - which ones would you guys like to see on your shelves? } } Thanks in advance } } Mike Gregory Professor Mike Gregory Electron Microscope Unit, University of Durban-Westville Private Bag X54001 Durban, Natal, South Africa 4000
(1) Jim Darley's method of two fix stages is one we regularly use with prints also. A small dish of "first fix" then ends up in the bottle for whoever collects it to recycle the silver, while the second larger one completes the job. The extra agitation involved also helps to eliminate "dead spots" where one print has lain over another.
(2) HOWEVER, there is a problem which also occurs, in that sometimes we get areas on prints that slowly turn yellow, or in extreme cases pinky-brown, on subsequent exposure to light. This seems to happens especially to bits that have buckled out of the developer, so that they were exposed to air for a longer time while still wet with developer. No amount of fixing will cure this. Can anyine tell me what (chemically) is going on?
(3) Kodak TMAX 100 film likes extra long fixing. I do like this film , but one does need to give it twice as long in the fix.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
This is beginning to look like a chain letter but I thought I should add my two (UK) pence worth.
I have worked in a lab where we used a modified separating funnel to control the thickness of plastic coatings. This is the sort of gadget that you would use to separate organic solvent layers by draining them through a glass tap at the bottom. There is a type which has a cylindrical chamber which is just the right height and diameter to adequately cover a standard 76mm x 26mm glass slide. The modification was to take off the narrow neck (apparently a fairly straight-forward glass cutting exercise) so that slides could easily be placed in the chamber of the funnel.
The technique was to put the slide in the funnel and cover with the plastic solution. Let it settle to avoid any lumps or bubbles and then turn the glass valve at the bottom and drain the plastic into its stock bottle or a beaker. The beauty of this is that you can vary the flow rate of the plastic off the slide (fast for thick and slow for thin) which is much easier than controlling the rate of removal of a slide from the plastic. This often means that you could get much thinner and more even layers with careful selection of plastic, solvent and concentration (yes you do need to experiment). You can even get thicker layers by pulling the tap out to rapidly drain off the plastic.
Of course, do this in a fume hood.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
Disclaimer - it's not my invention and you pays your money and takes your chance. ----------
} From: Chris Gilpin To: Microscopy-at-Sparc5. Microscopy.
-----Original Message----- } From: Chris Gilpin [mailto:cgilpin-at-fs1.sem.man.ac.uk] Sent: 10 October 1998 13:14 To: Me at home
-----Original Message----- } From: Jim J Darley Sent: 10 October 1998 13:43 To: 'Garber, Charles A.'; MICROSCOPY BB Cc:
On Saturday, 10 October 1998 6:28, Garber, Charles A. [SMTP:cgarber-at-priam.chesco.net] wrote: . . . .There was some disagreement between telling all our "secrets" vs. not telling all our secrets. But here are the "secrets" which perhaps others might find of value. . . . The message is clear: Chuck & Co know how to make filmed grids better than anybody else. Sadly, I learned nothing from the "secrets", I wonder in fact: what were the secrets?
. . . . What is not generally appreciated is that there are a number of different grades of the resin called Formvar, but so far as we know, only one of them is optimum for the filmed grid application. If one was using other grades, they might require different concentrations for the best results. The grade that we use is the grade that we sell for this purpose. And no, we won't tell that little secret. . . . . Oh, now I learn that I was just lucky that all of the films that I had made over about 30 years worked well, yet none of the formvar came from Chuck. But why his reply? The questioner had no problems with formvar but was interested in technique to control thickness.
. . . . .But one really does need a TEM right there, next to where the films are being made, so that they can be instantly inspected and corrective action taken, right there on the spot, e.g. by way of a concentration change. If this is not done, the the QC step end up being when you gear up to do your experiment and then you find out the grids are not stable. .. . . . Other suppliers know nothing and get their films probably made at the back of a soup kitchen. You take a grave risk if you buy your films anywhere else. But why? The question had been on how to control film thickness and not were to buy filmed grids.
. . . .Disclaimer: SPI Supplies has produced custom coated filmed grids for TEM for customers worldwide for some number of years. More information about the coating of TEM grids can be found on our website given below. . . . . This is not a disclaimer, but another vintage, bold-faced advertisement by Chuck. Nobody had asked about his filmed grids and he gave no useful information on how to control film thickness well. Two other writers provided methods ("double dipping" and "lengths of time after dipping in solvent saturated atmosphere") that he may find useful to apply to his assembly line. He is too cheap to pay for advertising his filmed grids and has again used the microscopy server instead.
Non italics by Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
Dear list Jim Darley has a point. I was asking about ways of controlling film thickness. Here follows the background to my request which will clarify further...
I have a "customer" in my core facility embarking on a serial sectioning project. She needs to use slot grids and has chosen the .75 variety. For whatever reason the films are not lasting as far a prolonged viewing in the microscope. Yes we are carbon coating. Yes we have tried carbon coating before and after section collection. Yes we have tried glow discharging with and without carbon. If anything our films are a bit on the thin side (great for some applications but not this one!) We have produced some thick films but they are very prone to wrinkling. Now you will see the reason for the question. I would like to make thicker films than silver/gold but without wrinkling. Perhaps I am guilty of asking the bare question without detailing why. I would add however that all replies are greatly received. Members of the list never cease to surprise me with their willingness to help. Other posts to the list show that most of us are fighting for our lives to stay in "business". As a community we stand a better chance of survival if we are all successful. Sharing knowledge is part of making our labs a success. I appreciate that commercial companies have a living to make and can be forgiven for keeping trade secrets. As long as the rest of us don't fall into that trap.
Again many thanks to ALL who replied. Long live EM
Chris
Experimental Officer Biological Sciences EM Unit University of Manchester Oxford Road Manchester M13 9PT Phone +44 (0)161 275 5170 Fax +44 (0)161 275 5171
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I understand that the S1 accelerator used for curing Spurr's has a relatively short shelf life (I usually replace mine after about 6 months or a year). Also the anhydride hardener - NSA - is particularly prone to absorbing moisture so if a stock bottle has been opened for a long period and I have a problem I try an unopened one. In any event I always label bottles with the date of arrival in the lab and the date of opening.
Some moisture may be driven out of the resin during polymerisation but not if you seal the lids on polythene BEEM capsules.
How good is your temperature measurement? We have an old oven with a dial setting that needs to be checked if anyone else has used it and often the only way is to start polymerisation a couple of hours before I leave the lab on a night, so that I can confirm the polymerisation temperature.
I hope this helps.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
The usual disclaimer - these are my opinions and may have little basis in science or even fact. ---------- } From: Seung-Geuk Shin To: Microscopy ListServer
I had the same problem, when I used an old batch (3-5 years with occasional uses), or one exposed to moisture (If you keep the bottles in a low temperature for extended shelf life, let it warm before opening the bottle). With bottles out of the box, I had no problem. In my opinion, Spurr's has relatively poor cutting properties, even when the block itself is not brittle. I usually successfully mix Spurr's (complete homogeneous mixture including DMAE) with Epox 812 (complete homogeneous mixture including DMP-30) to improve cutting properties, with minimal sacrifice in Spurr's excellent viscosity. It's been very cooperative for plant tissues.
Seung-Geuk Shin SUNY-ESF at Syracuse sgshin-at-syr.edu
============================================ Disclaimer: The opinions expressed in this communication are solely my own. ============================================
} -----Original Message----- } From: Greg R [mailto:greg-at-umic.sunysb.edu] } Sent: Friday, October 09, 1998 11:11 AM } To: microscopy } Subject: Spurr resin } } Hi microsopists, } I am having a problem with Spurr resin. The } blocks are coming out very brittle. I am careful } when I weigh out the components and polymerize at } 65C. What else would cause brittle blocks? } Another problem is that staining with 1% T blue } with 1% borax is very poor. Any suggestions would } be of help. Thanks. } -- } Regards, } Gregory Rudomen } Technical Specialist } University Microscopy Imaging Center } State University of New York at Stony Brook } 516-444-3126 } Greg-at-umic.sunysb.edu } ********************************************************** } Standard disclaimer: The opinions expressed in this } communication are my own and do not necessarily } reflect those of the University Microscopy Imaging Center. } **********************************************************
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Hello all, Every message from Barbara Foster arrives with the heading but no text. Am I the only one this happens to?
Regards Eric ---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
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Doug,
Run, don't walk, to your nearest scientific library and go through the literature. Articles in current journals should give you a very good idea of the type of work being done by specific investigators at different Universities. Remember that microscopy is a technique that, unless you are in to instrument development, can be used to help solve many different questions. The area of cell biology is very broad. Try to find a copy fo the proceedings from the MSA meetings for the last year or two. Going through the abstracts may help you identify an area of particular interest.
My advise would be to find problems of interest to you which are being investigated with a broad range of techniques and can, or are presently, using microscopy as one of those techniques. It is very important for you as a graduate student to be exposed to many different techniques in this important phase of your training...although you may want to look for a laboratory with an emphasis in microscopy. There is plenty of time to narrow those techniques more in future post-doctoral positions.
When you identify a problem in cell biology that seems particularily interesting, you will be able to narrow your search for the right graduate program and primary investigator. Good luck with the search.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
School: Providence College Question: I've had so many questions answered through this service over the last 2 years - It's wonderful.
I'm a senior biology undergrad looking forward to continuing my education in grad school - hopefully a Ph.D program in cell biology. Who out there has opinions on institutions with exceptional microscopy facilities? I'm really into TEM/SEM but would love to experience some more advanced techniques and equipment. I really want a program that's interested in answerering questions of cell biology by pushing the limits of microscopy and related tools. Where should I look?
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Dear collegues,
We=B4re actually trying to establish an embedding method for = immunoelectronmicroscopy to detect epitopes by preserving fine = ultrastructure. In our first trial we had good results with LR-White resin and would = like to improve our protocol. To harden the blocks we add about 40 microlitres of accelerator to 10 ml = of the resin and then polymerize them at -35=B0C in UV-Light. But = although we do all the steps on ice it occurs that our new accelerator = works a bit to fast and the resin seems to polymerize superfast in a few = minutes developing quite good heat. Cutting the blocks we note = depremised that the specimen obviously did not catch any plastic. What can be done to avoid this? The accelerator is new and the resin is = about one year old. Anyone knows more about LR-White embedding? I would be thankful for any kind of reply. Bye, Michael
Michael Reiner Department of Anatomy University of Cologne (Germany) Joseph-Stelzmann-Str.9 50931 Cologne (K=F6ln)
Michael I have found that the recommended amount of accelerator (1drp/10ml) is way too hot for practical use. I experimented with 10 ml, 20,30 ml aliquots and 1 drp of accelerator, then polymerizing with temperature at -20C. I like 30 ml/drp of accelerator. If you need more cold try packing a little dry ice in the freezer to lower the temperature just a bit. (~5C). If you want to discuss further feel free to contact me. Marge
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
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Chris,
More details certainly permit more helpful responses. I did serial sectioning for years on large single hole grids using a very simple technique which made the problem of film thickness and wrinkles a very minor one. Even though this method does not deal directly with ways of determining film thickness, it may help you out with the specific problem at hand. I do not remember who originally gave me the idea for this method but I was not the developer. It goes as follows:
1) Have your machine shop cut some thin pieces of plexiglass into the size of glass slides. At one end, drill about a dozen holes, roughly 5mm in diameter in an area about the size of a formvar film cast on glass slides. These slides will serve as your template for holding your films.
2) Cast the formvar films on glass slides just like you normally do. Usually a good silver film, not grey, will work fine. I routinely used 0.2% in ethylene dichloride when casting by immersing the slide into the solution in a small jar, etc. We now use a film caster which lets us hold the slide in the dichlorothane vapors after lowering the formvar solution level. I would have to redetermine correct percentage and timing since this method tends to give you thinner films consistantly.
3) Float the film off the glass slide and pick it up with the plexiglass slide so the film covers the holes. Then draw the water out of the holes by pressing the plastic slide down onto filter paper, or using small pieces of filter paper and capillary action to draw the water out of individual holes. Even thin films will hold nicely over the holes in the slide. Store slides until needed.
4) Next, cut your sections using a block diameter that is fairly similar to the size of the slit in the grid. Pick up the sections on UNCOATED grids by gently lowering the grid to the surface of the knife boat. I put the dull side down on the premise that the rough surface would grab the film better during step 6. The surface tension of the water will hold the sections in the grid opening. Transfer the grid to a droplet of water until you are finished sectioning.
5) Now transfer the grid + sections + water droplet to a drop of stain. Allow the section to stain, then wash by transferring through a series of droplets of clean water. Continue to post-stain if desired and wash the same way. Never let the grid dry.
6) Final step is to transfer the grid to a film suspended over the hole in a plexiglass slide and let it dry down. The sections will now be stuck to the film with NO wrinkles and minimum breakage. Also there was minimum problem with stain percipitation, but you must use very clean water. When ready to view, just punch out around the grid with the tip of your forceps, grab the grid and insert into the mciroscope.
Believe me....the sections will still be there at the end!!!
I found that as long as the sections cover a substantial portion of the open area of the grid, carbon coating was not essential. I used to do 50-100 grids worth of serial sections without loosing any. The films on the plastic slides would hold for months so I could make a lot and store until needed. The method really works...do give it a try.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
P.S. Taking the time to write this stuff down has turned out to be very helpful.....it goes into a technique notebook for future users!! --------------------------------------
Dear list Jim Darley has a point. I was asking about ways of controlling film thickness. Here follows the background to my request which will clarify further...
I have a "customer" in my core facility embarking on a serial sectioning project. She needs to use slot grids and has chosen the .75 variety. For whatever reason the films are not lasting as far a prolonged viewing in the microscope. Yes we are carbon coating. Yes we have tried carbon coating before and after section collection. Yes we have tried glow discharging with and without carbon. If anything our films are a bit on the thin side (great for some applications but not this one!) We have produced some thick films but they are very prone to wrinkling. Now you will see the reason for the question. I would like to make thicker films than silver/gold but without wrinkling. Perhaps I am guilty of asking the bare question without detailing why. I would add however that all replies are greatly received. Members of the list never cease to surprise me with their willingness to help. Other posts to the list show that most of us are fighting for our lives to stay in "business". As a community we stand a better chance of survival if we are all successful. Sharing knowledge is part of making our labs a success. I appreciate that commercial companies have a living to make and can be forgiven for keeping trade secrets. As long as the rest of us don't fall into that trap.
Again many thanks to ALL who replied. Long live EM
Chris
Experimental Officer Biological Sciences EM Unit University of Manchester Oxford Road Manchester M13 9PT Phone +44 (0)161 275 5170 Fax +44 (0)161 275 5171
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RECEIVED: from SF_Database by POP_Mailbox_-1304058404 ; 11 OCT 98 00:01:32 UT Received: from SPARC5.MICROSCOPY.COM by mailcenter.btny.purdue.edu with SMTP (QuickMail Pro Server for MacOS 1.0.3); 11 OCT 98 00:01:15 UT Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id QAA10063 for dist-Microscopy; Sat, 10 Oct 1998 16:28:20 -0500 Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id QAA10060 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Sat, 10 Oct 1998 16:27:49 -0500 Received: from gannet.scg.man.ac.uk (gannet.scg.man.ac.uk [130.88.94.110]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id QAA10053 for {Microscopy-at-sparc5.microscopy.com} ; Sat, 10 Oct 1998 16:27:36 -0500 Received: from fs1.sem.man.ac.uk ([130.88.91.100]) by gannet.scg.man.ac.uk with esmtp (Exim 1.92 #2) for Microscopy-at-Sparc5.Microscopy.Com id 0zS6hZ-0003X8-00; Sat, 10 Oct 1998 22:37:25 +0100 Received: from MAN-SEM-FS1/MAILQUEUE by fs1.sem.man.ac.uk (Mercury 1.21); 10 Oct 98 23:09:48 GMT Received: from MAILQUEUE by MAN-SEM-FS1 (Mercury 1.21); 10 Oct 98 23:09:20 GMT Received: from blackpig by fs1.sem.man.ac.uk (Mercury 1.21); 10 Oct 98 23:09:12 GMT From: "Chris Gilpin" {cghome-at-fs1.sem.man.ac.uk} To: "Microscopy-at-Sparc5. Microscopy. Com" {Microscopy-at-Sparc5.Microscopy.Co m} Subject: RE: RE: Formvar (R) coating of TEM grids Date: Sat, 10 Oct 1998 22:36:02 +0100 Message-ID: {000601bdf495$fa29f4c0$e65b5882-at-blackpig} MIME-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: 7bit X-Priority: 3 (Normal) X-MSMail-Priority: Normal X-Mailer: Microsoft Outlook 8.5, Build 4.71.2173.0 Importance: Normal X-MimeOLE: Produced By Microsoft MimeOLE V4.72.3110.3 In-Reply-To: {01BDF491.A83F5500.jim-at-proscitech.com.au} Errors-to: Microscopy-request-at-sparc5.microscopy.com
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We have used Animal Pharm for years with good results.
Animal Pharm Services 7711 West Dry Creek Road Healdsburg, CA 95448 USA
(707) 431-0171, (800) 808-0550, FAX (800) 808-0551 Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
Dr Eric E. Lachowski wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello all, } Every message from Barbara Foster arrives with the heading } but no text. Am I the only one this happens to? } } Regards } Eric } ---------------------- } Dr Eric E. Lachowski } University of Aberdeen } Department of Chemistry } Meston Walk } Old Aberdeen AB24 3UE } Scotland } +44 1224 272934 } e.lachowski-at-abdn.ac.uk
I sent out some messages last week and they arrived with a heading but no content.
ELECTRON MICROSCOPY TECHNICIAN The Integrated Microscopy Core, Department of Cell Biology, Baylor = College of Medicine has an immediate full-time opening for an electron = microscopy technician. The Integrated Microscopy Core is a busy, = state-of-the-art facility with 2 TEMs, deconvolution, laser scanning = confocal and 2 CCD-based fluorescent microscopes. The applicant should = have at least 1-2 years experience in all aspects of sample preparation = for biological TEM including fixation, embedding, ultrathin sectioning = and staining of tissue samples and cell monolayers. The applicant = should have darkroom experience and some experience in the operation of = TEMs. Immunolabelling experience is desirable but not essential. Other = duties include preparation of solutions, embedding media and the = maintaining of records. The position offers opportunities for training = in advanced light microscopy techniques, including fluorescence, laser = scanning confocal and deconvolution microscopy. The position requires a = minimum of a Bachelors degree and will start as a Lab Technician II; = salary range is low 20's, commensurate with experience, and includes the = standard Baylor benefits package.
Send CV and letter of research/technical interests to:
Hank Adams Laboratory Manager Integrated Microscopy Core Department of Cell Biology Baylor College of Medicine One Baylor Plaza Houston, TX 77030 Email submissions to: hpadams-at-bcm.tmc.edu Fax submissions to: 713 790 0545
Baylor College of Medicine is an Equal Opportunity, Affirmative Action = and Equal Access Employer.
Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
Here is the recipe for Spurr's + Epox 812, I use. Basically, it is 1:1 mixture (by weight) of Spurr's and Epox 812. Each resin mixture must be completely homogenized, including accelerator, before combining the two.
1. Prepare Spurr's in a 100 ml disposable beaker. (Based on Spurr's original formula)
VCD -------- 10 g DER 736 ---- 6 g NSA -------- 26 g DMAE ------- 0.4 g ------------------- Total ------ 42.4 g
2. Prepare Epox 812 in a 50 ml disposable beaker. (Based on Luft's original formular, except for reduced DMP-30)
Epox 812 (WPE = 155*) --------- 25 g DDSA (MW = 266) -------------- 14 g NMA (MW = 178) -------------- 11 g DMP-30 ----------------------- 0.5 g
*If WPE is different, proportions of each component should be recalculated. Calculations can be found at http://www.emsdiasum.com/ems/techdata/68.html).
3. To make 1:1 mixture, add 42.4 g of Epox 812 mixture into the 100 ml beaker containing Spurr's and mix throughly. Amount of Epox mixture can be reduced for hard-to-infiltrate tissues.)
4. Infiltration for plant tissues.
Transitional solvent Resin Mixture Infiltration (propylene oxide or Time (with diethyl ehter) rotation or occasional swirring) -------------------- -------------- ------------ 3 part 1 part 2 hrs. 2 2 2 hrs. 1 3 4 hrs. 0 4 4 hrs. 0 4 6 hrs. -----------------------------------------------------------------
5. Cure blocks at 60 C for 48 hrs.
Regards,
Seung-Geuk Shin
} -----Original Message----- } From: Michelle L. Peiffer [mailto:mlk101-at-psu.edu] } Sent: Monday, October 12, 1998 12:31 PM } To: Seung-Geuk Shin } Subject: RE: Spurr resin } } } I was quite interested to see this reply, we routinely use Spurr's with no } trouble in insect tissues and cell cultures. However recently we have a } number of students without access to diamond knives who are having trouble } getting quality thin sections of plant tissue embedded in Spurr's. Would } you mind sharing your recipe for this spurr's epox mixture? Thanks . } } } #################################################### } Michelle Peiffer } Electron Microscope Facility for the Life Sciences } The Biotechnology Institute for Research and Education } 1 South Frear Lab } University Park, PA 16802 } 814-865-0212 email:mlk101-at-psu.edu } #################################################### } } }
The Australian Museum has purchased a Wentzscope for our new Biodiversity Gallery and we ordered a slide carrier from Henry Frickel, 124 N Janney St Baltimore MD 21224. We would be most interested to find out more about him, especially since he has our money and we have nothing! Has anyone had dealings with this person?
Hi Eric, it doesn't occurs with the Barbara Foster's messages that I receive. I don't know what happen but try open the message and click the "return to the author"option; it shows to you the text of the message. I don't know what's the mistery. Rejane Galindo Universidade Federal Rural de Pernambuco Plant Morphology and Anatomy Recife. Pernambuco, Brazil
Rejane Pimentel Galindo Universidade Federal Rural de Pernambuco Av. Boa Viagem, 6592/602 51130-000, Recife, Pernambuco, Brasil ggalindo-at-elogica.com.br Fax (081) 441 4697
there was a test strip that you could get for checking the fixing capacity of fixer by its pH and silver content.. It was called Fixing Bath Test and was manufactured by E. Merck. The pack of 100, I am using now, was purchased in 1991 so with frugal use it can last for years.
A pack of 100 strips costs about 16 UK pounds from Agar Scientific (it may be available from other e.m. or photographic suppliers) but I think that 0.16 of a UK pound is a reasonable price to test the quality of fixer. I wouldn't use it on fresh fixer unless we had problems and would normally only use it on heavily used film fixer so the cost compared with losing a valuable film or replacing fixer too quickly is justified. Before this I used lightly fogged photographic film or paper to test my fixer but this only gives a basic idea.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
Disclaimer - I have no connection with any of the companies mentioned other than as a satisfied customer. ---------- } From: jim-at-proscitech.com.au To: 'Andy Burrows'; Microscopy-at-sparc5.microscopy.co
Andy: The fixer was too weak (or exhausted). It is a good idea to periodically check fixer. Place a small piece of undeveloped film in the fixer, full light does not matter. The film will become quite clear in under 30 seconds if its really good. If it takes a minute it's exhausted or made-up too weak. Exhausted fixer could be used to prefix film for a couple of minutes before using good fixer. This will prolong the good fixer's life. A brief water rinse after developing and before fixing is a good idea. I don not like acetic acid because with some materials it can cause mottling. Minimum fixation time is three times the time film takes to clear, but I suggest to not fix for less than three minutes. The colouration of under fixed materials only sometimes disappears when refixing. Certainly, underfixed images should be refixed if useful parts of the neg were unaffected and they too would change if not refixed promptly. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au
On Saturday, 10 October 1998 2:33, Andy Burrows [SMTP:ab0895-at-LIVERPOOL.AC.UK] wrote: } Hi to all, } I have just changed the solutions in our developer and fixer baths that serve our two TEMs. Unfortunately, the first couple of users } have informed me that their negatives are next to useless and that the negatives are pink around the edges! I'm pretty confident } that the developer is ok but the fixer put at my disposal (Agfa } Structurfix) was not what I used last time (Ilford Hypam). Any ideas? } Many thanks, } Andy } __________________________________ } Dr Andy Burrows } Materials Science and Engineering } Department of Engineering } The University of Liverpool } Liverpool } England } L69 3BX } } Tel: +(44) (0)151 794 5372 } Fax: +(44) (0)151 794 4675 } email: ab0895-at-liv.ac.uk
id xma029043; Tue, 13 Oct 98 08:56:44 -0400 Received: by eastman.com id AA78347 (5.67b/IDA-1.5 for Microscopy-at-Sparc5.Microscopy.Com); Tue, 13 Oct 1998 08:19:08 -0400 Received: from ntmcon02.emn.com by eastman.com with SMTP id AA39942 (5.67b/SMI-4.1 for {Microscopy-at-Sparc5.Microscopy.Com} ); Tue, 13 Oct 1998 08:19:06 -0400 Received: by ntmcon02.emn.com with Internet Mail Service (5.5.1960.3) id {4VN3HXB5} ; Tue, 13 Oct 1998 08:19:16 -0400 Message-Id: {E788D998AAC8D1119EC10000F8CD1E8A5B4189-at-ntmail23.emn.com}
I have no problem getting the text from Barbara's messages, although they are in a larger font (Arial 12pt) than most other messages (Arial 10pt).
Dennis B. Barr (dennbarr-at-eastman.com) Physical Chemistry Research Laboratory Physical & Analytical Chemistry Research Division Eastman Chemical Company Kingsport, TN 37662-5150
B-150B, R-132E, (423) 229-2188
} -----Original Message----- } From: Dr Eric E. Lachowski [SMTP:che136-at-abdn.ac.uk] } Sent: Monday, October 12, 1998 9:27 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: blank messages } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Hello all, } Every message from Barbara Foster arrives with the heading } but no text. Am I the only one this happens to? } } Regards } Eric } ---------------------- } Dr Eric E. Lachowski } University of Aberdeen } Department of Chemistry } Meston Walk } Old Aberdeen AB24 3UE } Scotland } +44 1224 272934 } e.lachowski-at-abdn.ac.uk } } }
OK - now I have a question about this...the fonts on the messages I get from Foster are always different and in bold, and MY terminal resets to that font about 90% of the time after a Foster message! Anyone know why that happens? It is very weird. If I then disconnect from the network, my terminal resets to "normal"..but I've never understood how she does this! (Not that it is intentional, but kind of a neat party trick)
Tamara "definitely not a network guru" Howard CSHL
On Tue, 13 Oct 1998, Barr, Dennis B wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have no problem getting the text from Barbara's messages, although } they are in a larger font (Arial 12pt) than most other messages (Arial } 10pt). } } Dennis B. Barr (dennbarr-at-eastman.com) } Physical Chemistry Research Laboratory } Physical & Analytical Chemistry Research Division } Eastman Chemical Company } Kingsport, TN 37662-5150 } } B-150B, R-132E, (423) 229-2188 } } } } -----Original Message----- } } From: Dr Eric E. Lachowski [SMTP:che136-at-abdn.ac.uk] } } Sent: Monday, October 12, 1998 9:27 AM } } To: Microscopy-at-Sparc5.Microscopy.Com } } Subject: blank messages } } } } ---------------------------------------------------------------------- } } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } -. } } } } } } Hello all, } } Every message from Barbara Foster arrives with the heading } } but no text. Am I the only one this happens to? } } } } Regards } } Eric } } ---------------------- } } Dr Eric E. Lachowski } } University of Aberdeen } } Department of Chemistry } } Meston Walk } } Old Aberdeen AB24 3UE } } Scotland } } +44 1224 272934 } } e.lachowski-at-abdn.ac.uk } } } } } } } }
Dear all, has anybody an idea about solving the following problem with ou= r Hummer VII sputter coater: we're using routinely N2 for the gold -sputtering process. When = we start the program, the vacuum pump works well and reached the needed millitorr for further pr= ocessing, e. g. for the ignition point of the N- plasma. This becomes visible as a bluish light. = After that, normally, the digital display counts the coated thickness, but since a few weeks ago ,= it needed much more time (1 or 2 hours instead of 10 min.!) until the desired (20-30nm) thi= ckness was reached. But additionally, when we checked the probes, there isn't any gold layer = visible. At first, we bought a new target, but until today the problem is still unresolved... Bernward Laube Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=E4tsstrasse 25 Germany 33615 Bielefeld phone: 0521 1065592 fax: 0521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
I have had this happen with JB-4-- a long time ago. The problem turned out to be excess heating during polymerization. I use a heat sink of ice water arround the blocks, but I don't use a vacuum. Instead, I seal the tops of the molds (peel away) and place the blocks, ice water and all, into the refrigerator overnite.
Karen Pawlowski
On Sat, 10 Oct 1998, Cox, Elizabeth wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have the same problem with JB-4 Plus resin blocks. They are very brittle } and tend to shatter during trimming which is very irritating. They have } been left in a vaccum dessicator in vials without lids on. The blocks have } set with small bubbles on the underside, a problem I haven't previously had } using JB-4, not sure why? } } Liz } } } } }
Dear all, Several people have written about blank messages or weird effects with fonts on messages from certain subscribers. Maybe the main problem is that some people are sending email with styled text from one of a number of modern email systems but older emailers are somewhat limited in their ability to understand this formatting information (generally text size, font and colour). If you use Eudora Pro or Light 3.x or later, Microsoft Outlook Express or Netscape mail you should have no problems reading such mail but other older email packages may well have difficulties.
The question we should maybe be asking is "How acceptable is it to send email with text formattings on this listserver?" Modern email systems that send more than just plain text can produce nice looking messages, IF YOU CAN READ THEM. Using such features widely may discriminate against those who don't have the latest computer software, however, thus reducing the usefulness of this forum as a worldwide discussion medium. As for me, I sometimes use styled text if I know that the recipient will be able to read it, but for postings to general forums such as this one, I always just use plain text.
What do others think?
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, or: ianmaclaren-at-hotmail.com Birmingham B15 2TT, http://web.bham.ac.uk/I.MacLaren England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Can anyone help me find a short course in electron diffraction and similar techniques or suggest an alternate strategy?
A student here would like to learn more than I can teach her about analyzing her materials and interpreting her data. She is open to any possibility for now, though her options might be narrowed depending on the choices.
Thanks
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 FAX (831) 429-0146 jmkrupp-at-cats.ucsc.edu
Mike, I did a search on Frickel but could only find his address, 124 N. Janney Street, Baltimore, MD, 21224-1705 and his phone number, 410-342-5772. As a native of Baltimore, I'd hate for him to give it a bad name (or make it worse than it already is!), so here're a few url's that may help:
http://www.bbb.org (the Better Business Bureau) http://epfl2.epflbalto.org/citygov/police/police1.htm (five l's and one 1) (Baltimore City Police ).
Next time try what we used to do in Saudi Arabia. Find an acceptable disinterested intermediary(bank,lawyer,accountant,export firm) that will hold and release the payment once all is done and everyone is satisfied, especially with international transactions. It will save you alot of headache. Good luck. Winston. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supr. 10/13/98 12:15:54 PM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I agree that all messages should be in plain text.
I especially do not like messages sent as attachments that have to be opened. This seems to be happening more frequently of late. I automatically delete all such msgs, unopened.
Ron
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com IBM Analytical Services; http://www.chips.ibm.com/services/asg
Are you interested in Digital Imaging for Microscopy? If so, exciting thi= ngs are happening at Sound Vision Inc. The founders of the acclaimed Leaf Systems, the MicroLumina and other digital products have developed the SVMicro, a new high-resolution digital camera for the microscope for abou= t $2000.
? The camera mounts directly to the microscope via a c-mount and is tethe= red to your computer. ? The software allows instant archiving of images in a simple click and s= ave fashion. ? The SVMicro utilizes three shot RGB technology that gives you selectabl= e resolutions for various output file sizes up to 9MB. ? It is perfect for sending images across the Internet, documentation, archiving or high-resolution publication. ? The camera is available with ECP parallel port PC or a SCSI connection for MAC users. ? The same camera also takes great black and white pictures in a one shot mode. ? Able to do long exposures for most types of fluorescence without any hi= gh cost cooling system ? Able to rotate the green filter in front of the sensor to take one shot images ? Can save sequential shots to your hard drive without leaving the plugin.
To find out more about the SVMicro Digital Camera for the microscope, ple= ase visit our web site at www.soundvisioninc.com. You can also call Sound Vis= ion =92s sales department at 508-270-0044. Or send me an email at gwray-at-soundvisioninc.com.
Its because Barbara's messages are always HTML formatted, and too many people's mail reader software will only read plain text. On some Win95 systems running Internet Explorer 4.0 (and probably Win98 as well), this font could be carried over to other MS products.
------------------------------- Scott D. Ireland North American Sales Manager Media Cybernetics, LP "The Imaging Experts" Tel: 716.473.0222 Fax: 716.473.8048 Pager: 888.691.2492 scott-at-mediacy.com http://www.mediacy.com http://www.optimas.com -------------------------------
-----Original Message----- } From: Tamara Howard [mailto:howard-at-cshl.org] Sent: Tuesday, October 13, 1998 9:05 AM To: Barr, Dennis B Cc: Microscopy-at-sparc5.microscopy.com
OK - now I have a question about this...the fonts on the messages I get from Foster are always different and in bold, and MY terminal resets to that font about 90% of the time after a Foster message! Anyone know why that happens? It is very weird. If I then disconnect from the network, my terminal resets to "normal"..but I've never understood how she does this! (Not that it is intentional, but kind of a neat party trick)
Tamara "definitely not a network guru" Howard CSHL
On Tue, 13 Oct 1998, Barr, Dennis B wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have no problem getting the text from Barbara's messages, although } they are in a larger font (Arial 12pt) than most other messages (Arial } 10pt). } } Dennis B. Barr (dennbarr-at-eastman.com) } Physical Chemistry Research Laboratory } Physical & Analytical Chemistry Research Division } Eastman Chemical Company } Kingsport, TN 37662-5150 } } B-150B, R-(423) 229-2188 } } } } -----Original Message----- } } From: Dr Eric E. Lachowski [SMTP:che136-at-abdn.ac.uk] } } Sent: Monday, October 12, 1998 9:27 AM } } To: Microscopy-at-Sparc5.Microscopy.Com } } Subject: blank messages } } } } ---------------------------------------------------------------------- } } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } -. } } } } } } Hello all, } } Every message from Barbara Foster arrives with the heading } } but no text. Am I the only one this happens to? } } } } Regards } } Eric } } ---------------------- } } Dr Eric E. Lachowski } } University of Aberdeen } } Department of Chemistry } } Meston Walk } } Old Aberdeen AB24 3UE } } Scotland } } +44 1224 272934 } } e.lachowski-at-abdn.ac.uk } } } } } } } }
Ihave always used Zero Grade Argon for sputtering, so cannot directly address your problem - but...
Several things can cause lack of deposition (I may forget some)
Vacuum chamber leak, even if chamber vacuum looks ok. Sample outgassing, even if chamber vacuum looks ok. Contaminated sputtering gas Contaminated/dirty target (unlikely since you changed) Inappropriate vacuum level (unlikely unless your gauge is broken) Inappropriate voltage/current - Not mentioned in your message.
Woody White McDermott Technology, Inc.
------------------------
Dear all, has anybody an idea about solving the following problem with our Hummer VII sputter coater: we're using routinely N2 for the gold -sputtering process. When we
start the program, the vacuum pump works well and reached the needed millitorr for further processing, e. g. for the ignition point of the N- plasma. This becomes visible as a bluish light. After that, normally, the digital display counts the coated thickness, but since a few weeks ago , it needed much more time (1 or 2 hours instead of 10 min.!) until the desired (20-30nm) thickness was reached. But additionally, when we checked the probes, there isn't any gold layer visible. At first, we bought a new target, but until today the problem is still unresolved... Bernward Laube Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universitaetsstrasse 25 Germany 33615 Bielefeld phone: 0521 1065592 fax: 0521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
_She is sending embeded html code, not plain vanilla ascii text.
Woody
OK - now I have a question about this...the fonts on the messages I get from Foster are always different and in bold, and MY terminal resets to that font about 90% of the time after a Foster message! Anyone know why that happens? It is very weird. If I then disconnect from the network, my terminal resets to "normal"..but I've never understood how she does this! (Not that it is intentional, but kind of a neat party trick)
Tamara "definitely not a network guru" Howard CSHL
On Tue, 13 Oct 1998, Barr, Dennis B wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have no problem getting the text from Barbara's messages, although } they are in a larger font (Arial 12pt) than most other messages (Arial } 10pt). } } Dennis B. Barr (dennbarr-at-eastman.com) } Physical Chemistry Research Laboratory } Physical & Analytical Chemistry Research Division } Eastman Chemical Company } Kingsport, TN 37662-5150 } } B-150B, R-132E, (423) 229-2188 } } } } -----Original Message----- } } From: Dr Eric E. Lachowski [SMTP:che136-at-abdn.ac.uk] } } Sent: Monday, October 12, 1998 9:27 AM } } To: Microscopy-at-Sparc5.Microscopy.Com } } Subject: blank messages } } } } ---------------------------------------------------------------------- } } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } -. } } } } } } Hello all, } } Every message from Barbara Foster arrives with the heading } } but no text. Am I the only one this happens to? } } } } Regards } } Eric } } ---------------------- } } Dr Eric E. Lachowski } } University of Aberdeen } } Department of Chemistry } } Meston Walk } } Old Aberdeen AB24 3UE } } Scotland } } +44 1224 272934 } } e.lachowski-at-abdn.ac.uk } } } } } } } }
I have been asked about the possibility of examining a small electronic part using SEM. The first possible problem, which occurred to me, was the charging of any non-conductive surfaces. It now seems that this may be a small problem compared to what else was pointed out. The assembly also has a magnetic source of ~1000 gauss. My first guess is that examination of such a sample with SEM is not possible. Being optimistic, I did place a small wall magnet in the chamber just to observe the effect. The resulting image resembled a work by Dali. Just to make sure I was not wrong, I thought that this might be a good question to pose to the list to find if anyone else has experience with similar samples. Any replies, pro or con, of examination of such samples would be welcome.
Thanks *********************************************** Dave Strecker mailto:djstrecker-at-ra.rockwell.com Rockwell Automation/Allen-Bradley Phone: (440)646-3250 Component Engineering ND246 Fax: (440)646-3416 1 Allen-Bradley Dr. Mayfield Hts., Ohio 44124 USA ***********************************************
There were four rather good treatments of electron diffraction published a number of years ago. Since the phenomenon involved should not have changed much, these should still be useful, IF you can still find the books:
The first is: An Introduction to Electron Diffraction, by B. E. P. Beeston, Part II of Practical Methods in Electron Microscopy, Vol. 1. A. Glauert, Ed. North-Holland, American Elsevier,1972, ISBN 0-444-10404-6
The second is: 'Intrepretation of Electron Diffraction Patterns', by Andrews, Dyson &Keowon, Plenum Press, 1967 (Library of Congress Card No. 68-19540)
The third is: Ch. 15 by Alderson & Halliday "Electron Diffraction" in the book 'Techniques for Electron Microscopy' D. H. Kay, Ed., 2nd. Ed., F. A. Davis Publishers
The fourth: Chapts. 4 & 5 in 'Electron Microscopy of Thin Crystals' by Hirsch, et. al, Butterworths, 1967
The first is a thorough and detailed treatment covering several hundred pages and starting from very first principles. The third and fourth are descriptive and much shorter. The second contains a detailed instructions, tables of data, etc. for interpreting ED patterns.
While we're on the subject, here are a few other references that might be of some historical interest:
Structure Analysis by Electron Diffraction, by B. K. Vainshtein, MacMillan, 1964
Electron Diffraction, by Z. G. Pinsker, Butterworths, 1953
Theory & Practice of Electron Diffraction, by Thomson & Cochrane, 1939, MacMillan
Electron Microdiffraction, by Spence & Zuo. Plenum Press, 1992
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Your comments are very reasonable. However, I use an old text-only newsreader because it is the only on that permits me to maintain all my saved messages and pointers on the host machine while I access it from 3 or 4 other sites. All the newer readers download the messages and require me to carry around a news.src floppy to keep track.
I vote for plain text: it's always clear and readable.
Although I routinely take 0.5-1.5 um sections before ultrathin sections, I rarely keep and photograph these semithin sections. (Photons - who needs 'em?) However, right now I need to coverslip some Spurr's semithins, and I remember that one gets better results with a different thickness coverslip than the usual No. 1. Can anyone enlighten me, or offer suggestions for the best way to photograph Spurr's sections?
As always, I sincerely appreciate any responses!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
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The House Ear Institute, located in the downtown Los Angeles area, is a = non-profit research and education organization dedicated to improving the = lives of people who have hearing and hearing related disorders.
We are looking for a special person interested in biomedical imaging. The = ideal person would be someone with previous experience in hearing research = with training in histology, protein purification, cell or molecular = biology, aseptic techniques, electron microscopy (including SEM and = immunocytochemistry), data collection or data management. A bachelors or = masters degree, or equivalent laboratory experience is required. =
Candidates with a strong laboratory background not related to microscopy = but with an interest in learning imaging techniques are encouraged to = apply.
Please mail or fax resume with a letter of application to:
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I tried to sign up for the SIMS listserver at LISTSERV-at-FTMON.ARL.MIL = but recieved an error message in return. I think they changed it = sometime back but I cannot find the new address. Does anybody else = subscribe to this listserver, and if so have the new address? Thank you = very much in advance.
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Tina: The thickness of the coverslip is so important, most microscope manufacturers engrave the correct thickness on each objective! Some refer to the coverslip as the first lens in the path after leaving the specimen. Generally the correct thickness is 0.17. A 1 1/2 thickness coverslip has a thickness of between 0.16and 0.19 so it is usually the best choice. (A # 1 was a thickness of 0.13 to 0.17 so can be suitable if it is at the thick end of the range). With that wide of a variation, obviously not all manufacturers of # 1 1/2 coverslips are equal. The true anal compulsive can check this with a micrometer. The thickness assumes that there is an infinitely thin layer of mounting medium between the 0.17 glass and the coverslip. To achieve this, one needs to mount the section on the coverslip and not the slide. Doing this is truely the sign of the obsessed (I haven't done it in many months, honest). In the real world, one can just use a sparing amount of mounting medium and press firmly down (down allow any lateral motion or you will get funky waves in the medium). It is usually hard to appreciate the difference (by eye) the mistake of using a # 1 instead of a #1 1/2 but it is very obvious when a student uses too much mounting media. it is impossible to focus at high mags. If you are using an oil immersion objective, the optics are much more forgiving on coverslip thickness. Good luck, Tom
-------------------------------------. } } } Hi, all- } } Although I routinely take 0.5-1.5 um sections before ultrathin sections, I } rarely keep and photograph these semithin sections. (Photons - who needs } 'em?) However, right now I need to coverslip some Spurr's semithins, and } I remember that one gets better results with a different thickness } coverslip than the usual No. 1. Can anyone enlighten me, or offer } suggestions for the best way to photograph Spurr's sections? } } As always, I sincerely appreciate any responses! } } Aloha, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have a possible cause for all the confusion. Perhaps our friendly neighborhood sysop can offer his opinions. I have looked at some archived messages from Barbara and discovered that older messages came through fine, but lately messages arrive as an attached file which I must save to my hard drive and open with a word processor. I get the text below, which includes HTML tags for document formatting. I also noticed that the header info (not pasted in this message) indicated older messages were written with Netscape mail client ("Mozilla") with MIME content type set to "text/plain". New messages were sent using Eudora Pro and content type "text/enriched" which could explain the HTML tags and the fact that someone wrote they received Barbara's email with a larger font size.
Barbara, perhaps if you try setting content type to "text/plain" everyone could read the messages. Then again, I may be off in the weeds!
Jim Passmore Analytical Chemist Cryovac Division, Sealed Air Corp.
The Listserver rules specifically state not to send attachments and to send all messages in plain text. That judgement was made at the start of the listserver many, many moons ago and far in advance of the recent discussion.
Fonts/Colors etc are fine but they do not sure a sufficiently useful purpose in Email. They just cause headaches for some subscribers. These items belong on a WWW page and NOT imbedded in an Email message.
Please show courtesy to your colleagues and remove all such settings from any Listserver Email posting.
Dear All, The method I use to test my TEM film developer (Kodak Rapidfix) is to put two drops of saturated KI (potassium iodide) into a small beaker of the fixer dipped out of the fixing tank. If spent, the fixer will turn milky from the Ag I. If the beaker stays clear the fixer is still fixing silver. I've had a small bottle of KI on my shelf for many years and it is easy to make up.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
I recently read the postings relating to a vendor using this List in a somewhat roundabout way to advertise!
I have a proposal to make which I believe would put an end to this.
Vendors should all use the same simple disclaimer in their posts. This would read simply: "We are a vendor of electron microscope supplies" or words to that effect.
In addition, the body of the posting should address itself only to the specific questions being answered and not slip in statements like "...which this company sells....etc" unless someone is asking directly for a source.
We have the subject 3-chip, cooled CCD camera on a Nikon microscope with a Diagnostic Inst. adapter. When imaging a white field or any field with limited range in the histogram we like to expand the limited histogram to fill all 256 bins. When we do that we get a color distortion that, in a white screen gives pink on the top and green at bottom; middle is white. This does not happen with any of our single chip cameras.
We suspect that the color distortion is due to slight misalignment of the chips or prisms so that the R,G & B do not quite combine to give pure white. We have contacted the manufacturer, but they seem clueless, or suggest it is a microscope optics problem (of course!). We, would like to get this $25K camera working properly, or at least isolate the problem. Possibly 3-chip cameras cannot be "perfectly" aligned....we'd like to know.
Anyone out there have any experience with this kind of problem?
Dave Pevear, Exxon Production Research Co, Box 2189, Houston, TX 77252-2189
} From my point of view (as an EM service engineer), introducing a magnet into any area in an SEM can cause trouble. Even brief contact between a magnetic sample and EM stages and sample exchangers can cause them to become slightly magnetic themselves. Even if your stage is made of brass or aluminum, there are still setscrews and other mechanisms which might need degaussing if the imaging of the scope is affected.
Can you please explain why at least one of my messages was also received with no content? I don't use the same format as Barbara and, in this case sent the message privately.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Barbara, Eric, and others . . . } } I have a possible cause for all the confusion. Perhaps our friendly } neighborhood sysop can offer his opinions. I have looked at some } archived messages from Barbara and discovered that older } messages came through fine, but lately messages arrive as an } attached file which I must save to my hard drive and open with } a word processor. I get the text below, which includes HTML tags } for document formatting. I also noticed that the header info } (not pasted in this message) indicated older messages were } written with Netscape mail client ("Mozilla") with MIME content } type set to "text/plain". New messages were sent using } Eudora Pro and content type "text/enriched" which could explain } the HTML tags and the fact that someone wrote they received } Barbara's email with a larger font size. } } Barbara, perhaps if you try setting content type to "text/plain" } everyone could read the messages. Then again, I may be } off in the weeds! } } Jim Passmore } Analytical Chemist } Cryovac Division, Sealed Air Corp.
I would be interested in hearing from anyone who is working on remote = control of EM's across the Internet. We have people who are interested = in this and it would help greatly if we could get some pointers.
TIA
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} Hi, {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} I would be interested in hearing from anyone who is = working on=20 remote control of EM's across the Internet. We have people = who are=20 interested in this and it would help greatly if we could get some=20 pointers. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} TIA {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Colin {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Colin Reid, {BR} Electron Microscope=20 Unit, {BR} Trinity College Dublin, {BR} Dublin = 2, {BR} Ireland. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Tel: 353-1-6081820 {BR} Fax:=20 353-1-6770438 {BR} email: {A=20 href=3D"mailto:creid-at-tcd.ie"} creid-at-tcd.ie {/A} {/FONT} {/DIV} {/BODY} {/HTML}
Your comments on using a middle man/woman in commercial transactions are interesting. I for one would like to find out how the system works and also if people are using a similar system. I don't know if it is prevelant nowadays, but we personally have had two = bad deals in the last 24 months. We have always dealt on trust in the past = but this seems to have died out. First we placed orders for a TEM CCD camera and an LM CCD Camera with a Surrey based company ( Digital Pixel ). We were asked for and paid 30% = of the cost. 6-8 months later Dr Leslie Vanderpant attempted to install th= e TEM camera. He failed to get it to work and took it away. We never sa= w him, or the camera, again. We also did not receive the LM camera. Despite asking for a refund we have got nothing back and have lost approx. =A310K. Then I sourced a second-hand Microlumina in Carolina. We were asked for= a percentage before the camera would be shipped. Everything seemed fine until the individual absconded with the camera & the contents of the comp= any bank account. Thankfully his partner was an honest individual and he returned our money from his own bank account. These were two of our personal experiences recently, but it does raise th= e question of how safe your money is when you have to hand over large percentages to equipment companies. How do most people deal with this problem ( or have we just been unlucky = ? ) ?
Colin Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Winston Wiggins {wwiggins-at-carolinas.org} To: Michael Dingley {michaeld-at-amsg.austmus.gov.au}
Thanks to all who replied to my posting. Barbara has changed her settings and is coming through loud and clear.
Regards, Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
---------- } De: Ian MacLaren {I.MacLaren-at-BHAM.AC.UK} } Para: Microscopy-at-sparc5.microscopy.com } Assunto: Fonts in messages etc. } Data: Ter=E7a-feira, 13 de Outubro de 1998 12:30 } =20 } -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America= =20 } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } =20 } =20 } Dear all, } Several people have written about blank messages or weird effects with } fonts on messages from certain subscribers. Maybe the main problem is that } some people are sending email with styled text from one of a number of } modern email systems but older emailers are somewhat limited in their } ability to understand this formatting information (generally text size, } font and colour). If you use Eudora Pro or Light 3.x or later, Microso= ft } Outlook Express or Netscape mail you should have no problems reading su= ch } mail but other older email packages may well have difficulties. } =20 } The question we should maybe be asking is "How acceptable is it to send } email with text formattings on this listserver?" Modern email systems that } send more than just plain text can produce nice looking messages, IF YO= U } CAN READ THEM. Using such features widely may discriminate against tho= se } who don't have the latest computer software, however, thus reducing the } usefulness of this forum as a worldwide discussion medium. As for me, = I } sometimes use styled text if I know that the recipient will be able to read } it, but for postings to general forums such as this one, I always just use } plain text. } =20 } What do others think? } =20 } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Ian MacLaren, Tel: (44) (0) 121 414 3447 } IRC in Materials for FAX: (44) (0) 121 414 3441 } High Performance Applications, email: I.MacLaren-at-bham.ac.uk } The University of Birmingham, or: ianmaclaren-at-hotmail.com } Birmingham B15 2TT, http://web.bham.ac.uk/I.MacLaren } England. } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } =20 } =20 } =20
Check out http://tpm.amc.anl.gov/mmc/ and links therein to the laboratories involved in the DOE2000 project on TelePresence Microscopy. Ours is http://www.ornl.gov/doe2k/, for example. Feel free to call for more information.
Larry
} Hi, I would be interested in hearing from anyone who is working on } remote control of EM's across the Internet. We have people who are } interested in this and it would help greatly if we could get some } pointers. TIA Colin Colin Reid, } Electron Microscope Unit, } Trinity College Dublin, } Dublin 2, } Ireland. Tel: 353-1-6081820 } Fax: 353-1-6770438 } email: creid-at-tcd.ie
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
The correct coverslip thickness for a given objective is engraved on its mount. Keep in mind, that this "coverslip thickness" is the sum of:
thickness of adhesive+thickness of the section/specimen+thickness of the mounting medium+thickness of the coverslip...
In that respect, "preparation thickness" is more correct!
To thick a preparation results in undercorrection of spherical abberation. This is usually not a big problem with objectifs NA 0.40 or less and visual observation. It is a problem with dry objectives of higher NA and in photomicrography: the image looks very "blured", details are missing...
This can be easily demonstrated: take a slide and put a second coverslip on it with a small drop of paraffin- or immersion oil and examine, you'll notice the effect...
Immersion objectives are much more forgiving on coverslip thickness, due to the fact that there are little or no deviations in refracting index between specimen and front lens of the objective ("homogenious immersion").
Keep in mind however, that the working distance of a 100x immersion lens is only about 0.1mm...
As some kind of rule of thumb:
* with an NA 0.90 (dry), deviations in "coverslip thickness" of 1 - 2=B5m ar= e noticeable... * with an NA 0.50, deviations in "coverslip thickness" of 50=B5m are not noticeable... (James, J.J.:"Microscopische waarnemingsmethoden", Oosthoek, Utrecht, The Netherlands, 1969).
This may be too simple, but what room lighting are you using while grabbing images? I get the same kind of thing with an ordinary single chip camera with fluorescent room lights on. It's especially strong at low magnifications when more stray light enters the objective. Hope this helps.... -- James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
We have four Hitachi 3-chip color cameras (HV-C20) with couplers from both Diagnostic Instruments and Optem International. These cameras have a shading correction function which attempts to even out the red to green color shading (this appears to be a universal problem with 3-chip cameras). I have not noticed the problem as much when the cameras are used with conventional video camera lenses.
On the other hand, I have a problem with color fringing in certain parts of the images. It is most noticeable with high contrast images, such as black lines on a white background. We have four of these cameras and they all have this problem to one degree or another. This particular problem also exists when using video camera lenses. It could be chromatic aberration, but I think it is a phenomenon of 3-chip cameras.
Any other ideas out there?
Dennis B. Barr (dennbarr-at-eastman.com) Physical Chemistry Research Laboratory Physical & Analytical Chemistry Research Division Eastman Chemical Company Kingsport, TN 37662-5150
B-150B, R-132E, (423) 229-2188
} -----Original Message----- } From: P00bare [SMTP:p00bare-at-pdq.net] } Sent: Tuesday, October 13, 1998 11:44 PM } To: Microscopy Listserver } Subject: Hamamatsu C5810 Camera } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } We have the subject 3-chip, cooled CCD camera on a Nikon microscope } with a } Diagnostic Inst. adapter. When imaging a white field or any field } with } limited range in the histogram we like to expand the limited histogram } to fill } all 256 bins. When we do that we get a color distortion that, in a } white } screen gives pink on the top and green at bottom; middle is white. } This does } not happen with any of our single chip cameras. } } We suspect that the color distortion is due to slight misalignment of } the } chips or prisms so that the R,G & B do not quite combine to give pure } white. } We have contacted the manufacturer, but they seem clueless, or suggest } it is a } microscope optics problem (of course!). We, would like to get this } $25K } camera working properly, or at least isolate the problem. Possibly } 3-chip } cameras cannot be "perfectly" aligned....we'd like to know. } } Anyone out there have any experience with this kind of problem? } } Dave Pevear, Exxon Production Research Co, Box 2189, Houston, TX } 77252-2189
I am looking for a software (database) where input would be (elemental) chemical composition of an unknown phase (from quantitative EDS and/or WDS analysis) and the output: possible minerals that match (inside some error boundaries)that chemical composition.
Very best regards,
Goran Drazic J. Stefan Institute Ljubljana Slovenia
I once saw a wonderful teaching tool for EM which consisted of a column which had been bisected longitudinally to show the windings in the lenses, etc. I have an old column, access to a good metal cutting bandsaw, and would like to do the same, but wonder if it's really feasible.
Does anyone know if there are any special tricks to doing this?
I'm pretty sure Barbara did not do anything intentionally. I know here at Cal the computer people change things all the time without telling anybody, and us computer "illiterates" keep doing what we've always done not knowing we're doing something wrong. I never had a problem with her messages, but at least she got us all to pay attention to how our e-mail systems are configured so we can make sure stuff is sent out correctly. Thanks Barbara for bringing this to all of our attentions! I'm checking all my settings right now.
Still trying to figure out what HTML stands for (looks like hatemail to me),
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
I apologize for the tone but I feel that this was a terrible answer to send out. I assume that the person who asked the question is either young or not too familiar with the ideas of science. Therefore to imply that the magnification requested can be known so accurately (more accurately than we know any of the fundamental constants, for example) is leading the reader astray, to put it mildly. If the diameter of a quarter varies by, say, a part in a thousand from one to another, the answer would be 574 times ten to the power six i. e. 574 million. All the other figures are wrong.
Alwyn Eades
At 09:04 AM 10/14/98 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Dear Dave, I have examined a cobalt-based super-magnet in the SEM, but strictly for EDX analysis. The image is grossly distorted. The only suggestion would be to use as long a working distance as possible and as high an accelerating voltage. Near the magnet will still be distorted. You wrote: } } I have been asked about the possibility of examining a small electronic } part using SEM. The first possible problem, which occurred to me, was } the charging of any non-conductive surfaces. It now seems that this may } be a small problem compared to what else was pointed out. The assembly } also has a magnetic source of ~1000 gauss. My first guess is that } examination of such a sample with SEM is not possible. Being } optimistic, I did place a small wall magnet in the chamber just to } observe the effect. The resulting image resembled a work by Dali. } Just to make sure I was not wrong, I thought that this might be a good } question to pose to the list to find if anyone else has experience with } similar samples. Any replies, pro or con, of examination of such } samples would be welcome. } } Thanks } *********************************************** } Dave Strecker mailto:djstrecker-at-ra.rockwell.com } Rockwell Automation/Allen-Bradley Phone: (440)646-3250 } Component Engineering ND246 Fax: (440)646-3416 } 1 Allen-Bradley Dr. } Mayfield Hts., Ohio 44124 USA } *********************************************** Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Madam or Sir, My name is Hong Feng Zhang. I am a new member of the Microscopy Society of America. I saw the other member got the email message from Listserver of MSA, such as EM job opening. I would be very appreciated if you could tell me some information about how to get on to the list. I am interested in looking for an EM technician position in NY. Is there any way I could post this message to the other member of our Society?
Thank you very much for your consideration. I am looking forward to hearing from you soon.
Cordially, Hong Feng Zhang Phone: (718) 319-8240 Email: hfzhang-at-aecom.yu.edu
And seconds to the empathy expressed to Barbara. (After my own experience with "Gold" mishap a few weeks ago, I know what she must be going through.)
Fortunately, the majority of subscribers on this list are quite understanding professionals who've got better things to do than keep up with the idiosyncrasies of computer networks. And we want to continue to hear from them. We would do ourselves a great disservice in discouraging their input by forcing our own standards of computerese upon them. In that case, we would lower ourselves to the level and thinking of those rednecks who stand up against U.S. immigrants on the basis of their not having a mastery of the English language. There are plenty of places for computer professionals (newsgroups on the USENET, for example), and those intent on debating these kinds of issues should exit to those; on the other hand, there are not enough forums for people like Barbara and Paula, and we should be working to make the few such as this that are available as friendly as possible.
Paula Sicurello wrote: } } Listers, } } I'm pretty sure Barbara did not do anything intentionally. } I know here at Cal the computer people change things all the time } without telling anybody, and us computer "illiterates" keep doing } what we've always done not knowing we're doing something wrong. } I never had a problem with her messages, but at least she got us } all to pay attention to how our e-mail systems are configured so } we can make sure stuff is sent out correctly. } Thanks Barbara for bringing this to all of our attentions! } I'm checking all my settings right now. } } Still trying to figure out what HTML stands for (looks like hatemail } to me), } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } phone: 510-642-2085 } fax: 510-643-6207 } http://biology.berkeley.edu/EML
-- ..we now return control of your computer screen to you... ---------------------------------------------------------- Timothy G. Moeller | Microanalysis Products Senior Software Engineer | NORAN Instruments Inc., {tmoeller-at-noran.com} | a ThermoSpectra company ----------------------------------------------------------
Colin, Try contacting an import/export company first. Make sure they're experienced in the country in which you're making your purchase. Give them the details and ask if they will negotiate or if they have a standard procedure to act as intermediary. Make them aware of your previous experiences. This is a technique we commonly used in the Middle East to make sure that products were delivered or money was returned, no slander intended. If the selling party refuses to agree to the intermediary, that should raise a red flag for you. Good luck. Winston.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supr. 10/14/98 12:56:23 PM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I need to know the diameter of a quarter and the diameter of the world. } How many times a quarter would have to be magnified to cover the world?
At 08:24 AM 10/14/98 -0700, Paula Sicurello wrote: } Still trying to figure out what HTML stands for (looks like hatemail to me),
HTML=HyperText Markup Language. It is what web pages are written in. There are tags that identify what fonts, effects, or other special things to use. Similar codes are embedded in word processor documents. However, they are usually not readable if you were to look at your document with a text editor, whereas HTML tags are readable albeit a foreign language.
HTML allows for the fancy effects in a web brower or enhanced e-mail program, but sure looks ugly in a text editor or regular mail program.
What happens when you take the camera off the microscope and image a white sheet of paper? You may not even need a focused image or even the adapter. I suspect that you will get the same effect and go back to the camera maker and remake your case.
Of course, it may be that they come back and say that the colors were balanced within their specifications until you drastically rescaled the brightness.
Warren
At 09:43 PM 10/13/98 -0600, you wrote: } } We have the subject 3-chip, cooled CCD camera on a Nikon microscope with a } Diagnostic Inst. adapter. When imaging a white field or any field with } limited range in the histogram we like to expand the limited histogram to fill } all 256 bins. When we do that we get a color distortion that, in a white } screen gives pink on the top and green at bottom; middle is white. This does } not happen with any of our single chip cameras. } } We suspect that the color distortion is due to slight misalignment of the } chips or prisms so that the R,G & B do not quite combine to give pure white. } We have contacted the manufacturer, but they seem clueless, or suggest it is a } microscope optics problem (of course!). We, would like to get this $25K } camera working properly, or at least isolate the problem. Possibly 3-chip } cameras cannot be "perfectly" aligned....we'd like to know. } } Anyone out there have any experience with this kind of problem? } } Dave Pevear, Exxon Production Research Co, Box 2189, Houston, TX 77252-2189 } }
Did I get the attention of all you filmmakers with the title? Use butvar (EMS) either from powder or from liquid. It is MUCH stronger per thickness than formvar, and it has a lot less inherent "texture". You use it just like formvar. It is dissolved in chloroform. If anyone wants more information, let me know.
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i did this with an old philips column. to get a good clean segment i had to rough cut it (with a bandsaw) and then used a surface mill to "polish" it. some of the loose parts needed to be potted to hold them still during the milling. i kept the O-rings intact to show vacuum sealing, too.
it makes a wonderful "prop" for lectures on electron optics...good luck!
b-
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax) "Be well, do good work, and keep in touch"
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Don't forget that the sample is also part of the optical system. Best = results are obtained if the section is as close to the cover glass as = possible. We mount our sections directly onto the cover glass, not on the = slide.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org
with ESMTP id {01J2YR6PYPKM8Y7133-at-denver.du.edu} for Microscopy-at-MSA.Microscopy.Com; Wed, 14 Oct 1998 15:07:23 MDT Received: from localhost by du.edu (PMDF V5.1-10 #28062) with SMTP id {0F0U00H015CB6J-at-du.edu} for Microscopy-at-MSA.Microscopy.Com; Wed, 14 Oct 1998 15:07:23 -0600 (MDT)
Hi,
Spurr's is the most highly crosslinked of all embedding media used for LM-TEM. The higher the crosslinkage, the less likely any stain, whether for LM or heavy metal for TEM, is to penetrate well enough to be called successfull. First: Get rid of Spurr's. It contains a potent carcinogen, VCD. Do you need this? Second: If you cannot get rid of Spurr's (try real hard), then use the softest mixture you can tolerate for cutting. Polymerize at 60deg C overnight, and see if this is adequate. This will reduce the final crosslinkage. Third: Blocks in existence already! Don't stain well? Try soaking the sections prior to staining for extended periods of time in water, then 5 min in alcohol, in increasing concentrations until you reach 95%. Then go back down to water stepwise. Try staining. Fourth: Use as alkaline as possible a vehicle for your stain. pH 12 is about right. Fifth: Combine all of the above. Does not work? Soak the sections in water and gradually bring to 95% alcohol.. Expose to stain dissolved in alcohol. Does not work? Forget it. Start over.
If you have very valuable sections and you must stain them for TEM, use alcoholic UA for 10min at 60deg C. Use Reynolds lead citrate at a pH of about 9 or 10. This last trick is truly a last resort, since the lead may dump erratically (or stain easily) at this low pH.
If you polymerize a block at a low power for 45 minutes in a microwave, you crosslink the resin to such an extent that nothing, nothing, nothing you do will stain it. (Lost my best meat loaf dish this way). For TEM we do not polymerize resins totally, about 10% of the monomers are left unreacted with one another. If you "drive" the crosslinkage, the block will be harder, less elastic, and impenetrable for liquids (except if you boil it for a year or so in water). I am not exaggerating. I got this info out of one my very favorite materials science books.
Hi All: I'm looking for a independent service person who is experienced in repairing Cambridge SEM's. Preferably the Cambridge 120. Is there anyone like this in the Dallas-Fort Worth or Texas Area?
Regards,
Michael Coviello Materials Science UT Arlington
UT Arlington Attn: Michael Coviello Box 19031, 500 W. 1st Street Arlington, TX 76019
I have recently been involved with installing SEMs for equipment reseller= s. The industry policy is to have the equipment 100% paid before shipment. This presents some problems for both the buyer and seller as some buyers have = a company policy of paying for equipment "Net 30 days" after delivery. The = sellers have also had equipment stolen from them after the buyers made a partial payment.
Recently, I have attempted to resolve this problem by purchasing the equi= pment from the reseller AFTER I have received a purchase order from the Buyer. = After installation, the buyer (a large Company) informed me that they were not = paying for the remaining balance as the dollar amount that they had written on t= he purchase order "was an arbitrary amount to be changed as they saw fit". T= hey further informed me that they had attorneys that could drag this out for = as long as possible. I lost about $5,000.00 USD on the arrangement.
I now only deal with Sellers and Buyers I personally know and have done a reasonable amount of business. I now have a Buyer for an SEM that is givi= ng $80,000.00 USD to an equipment reseller. As the Buyer put it "this goes a= gainst every grain in my body; I DO NOT pay in full for equipment I have not ev= en seen but I trust you and you trust the Seller so we sent the check".
You are right, trust has everything to do with the transaction. I now onl= y do business with people I trust. The Asians have long chided the American wa= y of doing business as Americans do not want to establish a personal relations= hip with their clients. Perhaps they had it right all along.
Regards,
Earl Weltmer
Colin Reid wrote:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } } Winston, } } Your comments on using a middle man/woman in commercial transactions ar= e } interesting. I for one would like to find out how the system works an= d } also if people are using a similar system. } I don't know if it is prevelant nowadays, but we personally have had tw= o bad } deals in the last 24 months. We have always dealt on trust in the pas= t but } this seems to have died out. } First we placed orders for a TEM CCD camera and an LM CCD Camera with a } Surrey based company ( Digital Pixel ). We were asked for and paid 30= % of } the cost. 6-8 months later Dr Leslie Vanderpant attempted to install = the } TEM camera. He failed to get it to work and took it away. We never = saw } him, or the camera, again. We also did not receive the LM camera. } Despite asking for a refund we have got nothing back and have lost appr= ox. } =A310K. } Then I sourced a second-hand Microlumina in Carolina. We were asked f= or a } percentage before the camera would be shipped. Everything seemed fine } until the individual absconded with the camera & the contents of the co= mpany } bank account. Thankfully his partner was an honest individual and he } returned our money from his own bank account. } These were two of our personal experiences recently, but it does raise = the } question of how safe your money is when you have to hand over large } percentages to equipment companies. } How do most people deal with this problem ( or have we just been unluck= y ? ) } ? } } Colin } Colin Reid, } Electron Microscope Unit, } Trinity College Dublin, } Dublin 2, } Ireland. } Tel: 353-1-6081820 } Fax: 353-1-6770438 } email: creid-at-tcd.ie } -----Original Message----- } } From: Winston Wiggins {wwiggins-at-carolinas.org} } To: Michael Dingley {michaeld-at-amsg.austmus.gov.au} } Date: Tuesday, October 13, 1998 11:14 PM } Subject: RE: Help with slide carrier } } } ----------------------------------------------------------------------= -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of Americ= a } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.C= om } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht= ml } } ----------------------------------------------------------------------= -. } } } } } } } } Mike, } } I did a search on Frickel but could only find his address, } } 124 N. Janney Street, Baltimore, MD, 21224-1705 and his phone } } number, 410-342-5772. As a native of Baltimore, I'd hate for him } } to give it a bad name (or make it worse than it already is!), so } } here're a few url's that may help: } } } } http://www.bbb.org (the Better Business Bureau) } } http://epfl2.epflbalto.org/citygov/police/police1.htm (five l's and on= e 1) } } (Baltimore City Police ). } } } } Next time try what we used to do in Saudi Arabia. Find an } } acceptable disinterested intermediary(bank,lawyer,accountant,export fi= rm) } } that will hold and release the payment once all is done and everyone i= s } satisfied, } } especially with international transactions. It will save you alot of } headache. } } Good luck. } } Winston. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } Winston W Wiggins, Supr. 10/13/98 12:15:54 PM } } CRC-Electron Microscopy Lab. Ofc:704/355-1267 } } Carolinas Medical Center Fax:704/355-7648 } } P.O. Box 32861 Lab:704/355-7220 } } Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } } } }
There are a number of articlea on magnetic domain imaging. The idea is to keep the magnetic material away from the polepiece. As far as charging of the package goes one could go to a low KV or paint the non-conductive package surface with conductive paint.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dave, } } } From my point of view (as an EM service engineer), introducing a magnet into } any area in an SEM can cause trouble. Even brief contact between a magnetic } sample and EM stages and sample exchangers can cause them to become slightly } magnetic themselves. Even if your stage is made of brass or aluminum, there } are still setscrews and other mechanisms which might need degaussing if the } imaging of the scope is affected. } } Sincerely, } } Bill Carmichael } } Allequash Engineering
RESEARCH RESOURCES CENTER, UNIVERSITY OF ILLINOIS AT CHICAGO
Position Title: HEAD, ELECTRON MICROSCOPY FACILITY
The Research Resources Center (RRC), University of Illinois at Chicago, is seeking a microscopist to manage its Electron Microscope Facility, a UIC core instrumentation facility located in the UIC Health Sciences Center and serving primarily the biological and health-related sciences. The position carries the academic professional title Research Electron Microscopist. This individual will manage and participate in all EMF functions including: providing users with access to instruments, training, services and technical advice, and maintaining instruments. Other duties are supervising the EMF staff, recommending equipment acquisitions and upgrades; developing an annual budget; administrative tasks; methods development; and reporting to and advising the RRC Director. Professional development is encouraged.
The Health Sciences Center EMF contains five JEOL electron microscopes including a new 1220 TEM, and a new high resolution, Field Emission-6320F SEM; Topometrix STM/AFMs; and a Zeiss 510 LSCM with 3 lasers. Support features include off-line analysis and processing of data from any of the computerized microscopes, specimen preparation lab; ultramicrotomes, and darkroom. Personnel include two experienced EM technologists and a confocal microscopist.
Candidates must have: a doctorate and considerable postdoctoral, independent research experience utilizing electron microscopy; significant theoretical understanding of and skills in utilizing modern, computerized TEM and SEM instruments and related analytical techniques; application skills for imaging and analyzing a variety of specimens, and digital image processing of microscopy data. They should also have the necessary administrative, communication, and social skills for managing a centralized instrument service facility and working effectively with faculty, staff and students. In order to develop the facility's capabilities in high resolution, FE-SEM and in ultrastructural immunocytochemistry, significant expertise in these areas is desirable.
Further information may be obtained by consulting the following website: http://www.rrc.uic.edu/jobs.html
For fullest consideration, interested parties should send an application letter, complete curriculum vitae, and the names and addresses of three references within three weeks after appearance of this notice to:
EMF Head Search Research Resources Center, m/c 937 University of Illinois at Chicago 835 S. Wolcott Ave. Chicago, Illinois 60612-7341
Mailing address: Robert F. Loizzi, Ph.D. Director, RRC-West Research Resources Center (m/c 937) University of Illinois at Chicago 835 S. Wolcott Avenue Chicago, Illinois 60612-7341
TEL: (312) 996-7600, or -8748 FAX: (312) 996-0539
Campus address: Rm. E102, Medical Sciences Bldg, m/c 937
Once upon a time in a past life I had to have EDX done on a rare-earth cobalt magnet. After the SEM operator tossed me out of his lab, I used a hysteresisgraph to demagnetize my sample. Then I was allowed back in the lab and we were able to do the analysis.
Janet Rice MCC Senior Member Technical Staff rice-at-mcc.com 512-338-3266
There are three tricks I know of (from the old RCAs that we had at the now-defunct Center for EM at U. Illinois): 1) Cut in a vertical plane slightly off-center, at a right-angle to the line of sight when standing directly in front of the 'scope. The off-center help with saving the positions for grid-holders and other such pieces. Although grid-holders, apertures strips, etc. should be out of the column when sawn. 2) Drill entry & exit holes in the metal to be removed just below and above the lenses. Inject any good, transparent resin into the lenses. This will hold them in place when the column is sawn. Inject *carefully* to avoid air-bubbles. 3) Have 2 or 3 (or 4 or ...) extra EMs to practice on.
Caveat: I worked at the Center. This work was done *long* before I got there by a highly skilled machinist. What I related above is what I was told by others who had been around when the guy did the work.
Phil
} I once saw a wonderful teaching tool for EM which consisted of a column which } had been bisected longitudinally to show the windings in the lenses, etc. I } have an old column, access to a good metal cutting bandsaw, and would like to } do the same, but wonder if it's really feasible. } } Does anyone know if there are any special tricks to doing this? } } Paul Grover
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 oshel-at-terracom.net or poshel-at-hotmail.com
Dear Wil, } } There were four rather good treatments of electron diffraction published a } number of years ago. Since the phenomenon involved should not have changed } much, these should still be useful, IF you can still find the books: } [skip] I would add Cowley's Diffraction Physics to this list. Yours, Bill Tivol
Randy- This is a very complex question... The necessity to accommodate the students, research projects, outside users, industrial users, etc. becomes difficult when all groups often seem to share deadlines. Second is the complexity of cost recovery... According to Federal and State regulations, in this country, core facilities funded with tax revenues are not intended to compete with private commercial industrial labs. Determining various rate for each sector is also difficult. I have done this at two different labs in the past, and recommend to KEEP IT SIMPLE. Try to get a co-op type system, if possible, pre-pay to cover all the maintenance contracts paid, and then an additional sum for the basic supplies. Also make sure you have money budgeted for salary for personnel, you will find it essential to have competent staff. Try a salary survey of other labs which have a recharge system, it is a great starting place to establish your rates. -Mike
On Wed, 7 Oct 1998, Randy Mandryk wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } What is a good general criterion for evaluation of a core microscopy } facility in an academic situation? What kind of ranking priority may be } assigned to different operations, such as research and service, management } of the core facility, cost recovery (user fee), teaching commitment } (regular and short courses), publications, etc. } } A user fee is levied in most labs for use of major instruments. The sample } preparation methods may influence the amount of time a particular } instrument is in use, specially a TEM. What would be an average estimate, } in terms of percent time spent per year on TEMs, SEMs and Confocals in a } core facility? } } } } Randy Mandryk } Microtechnique Lab, Room CW 225T } EXT 3473, Biological Sciences } University of Alberta } } }
I am in search of breaking pads for an LKB 2078 Histo Knifemaker. Any suggestions?
David O'Neil tel: (902) 426-8258 National Research Council of Canada fax: (902) 426-9413 Institute for Marine Biosciences 1411 Oxford St. Halifax, Nova Scotia B3H 3Z1 Canada email: david.o'neil-at-nrc.ca
You could try Alex Greene (ablue-at-io.com) 512.282-5507 or Walter Protheroe (Corvos-at-aol.com) 281.879-7211.
I would be glad to repair the Cambridge but I'm afraid the airfare and travel time would be excessive.
Earl Weltmer
Mike Coviello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi All: } I'm looking for a independent service person who is experienced in } repairing Cambridge SEM's. Preferably the Cambridge 120. Is there anyone } like this in the Dallas-Fort Worth or Texas Area? } } Regards, } } Michael Coviello } Materials Science } UT Arlington } } UT Arlington } Attn: Michael Coviello } Box 19031, 500 W. 1st Street } Arlington, TX 76019
Dear collegues, thank you for all the interesting and diversificated hints for my = LR-White embedding procedure. Yesterday I started embedding samples in = LR-White without the accelerator/catalyst, both in the oven at 60=B0C as = well as at -20=B0C in the UV-fridge in order to compare the grade of = ultrastructural resolution. I=B4ll tell you about my findings anyhow ...
Finally I=B4d like to ask about the possibly risks of Lowicryl because = Debby Sherman proposed to try out Lowicryl HM20. A few months ago we = ordered this stuff but up until now nobody agreed in working with this = substance fearing possible risks on health and life so that the delivery = box remains not opened somewhere in our Lab ... :-( Anyone, especially Debby, could tell me about experiences in working = with Lowicryl (HM20/K4) and the REAL risk of allergies? Are there secure = working conditions to avoid hazard? I=B4d like to know whether it=B4s = worth trying out this or if it=B4s better to keep hands off.
With best regards, Michael
Michael Reiner Department of Anatomy University of Cologne (Germany) Joseph-Stelzmann-Str.9 50931 Cologne (K=F6ln)
I am looking for information for acquiring images from a Philips 501 SEM and a Philips 300 TEM. I have done some research, but what I have come across so far is very expensive (for our budget). Does anyone have any ideas or experience with this situation?
Dear colleagues, I am going to prepare sections of polmer materials with a ultramicrotome for the TEM observation. Any suggestions for the sectioning conditions and sample preparation procedues, such as staining...? The materials will be ABS, PET, Nylon, and polypropylene. I appreciate any kinds of help.
Yes but, Spurr's has very low viscosity and does not require propylene oxide as an intermediate solvent. PO is a very powerful carcinogen. Most people seem to regard PO as essential with other epoxy resins. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Thursday, 15 October 1998 7:07, HILDEGARD CROWLEY [SMTP:hcrowley-at-du.edu] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } Hi, } } Spurr's is the most highly crosslinked of all embedding } media used for } LM-TEM. The higher the crosslinkage, the less likely any } stain, whether } for LM or heavy metal for TEM, is to penetrate well enough } to be called } successfull. } First: Get rid of Spurr's. It contains a potent } carcinogen, VCD. Do you } need this? } Second: If you cannot get rid of Spurr's (try real hard), } then use the } softest mixture you can tolerate for cutting. Polymerize } at 60deg C } overnight, and see if this is adequate. This will reduce } the final } crosslinkage. } Third: Blocks in existence already! Don't stain well? } Try soaking the } sections prior to staining for extended periods of time in } water, then 5 } min in alcohol, in increasing concentrations until you } reach 95%. Then go } back down to water stepwise. Try staining. } Fourth: Use as alkaline as possible a vehicle for your } stain. pH 12 is } about right. } Fifth: Combine all of the above. Does not work? Soak } the sections in } water and gradually bring to 95% alcohol.. Expose to } stain dissolved in } alcohol. Does not work? Forget it. Start over. } } If you have very valuable sections and you must stain them } for TEM, use } alcoholic UA for 10min at 60deg C. Use Reynolds lead } citrate at a pH of } about 9 or 10. This last trick is truly a last resort, } since the lead may } dump erratically (or stain easily) at this low pH. } } If you polymerize a block at a low power for 45 minutes in } a microwave, } you crosslink the resin to such an extent that nothing, } nothing, nothing } you do will stain it. (Lost my best meat loaf dish this } way). For TEM we } do not polymerize resins totally, about 10% of the } monomers are left } unreacted with one another. If you "drive" the } crosslinkage, the block } will be harder, less elastic, and impenetrable for liquids } (except if you } boil it for a year or so in water). I am not } exaggerating. I got this } info out of one my very favorite materials science books. } } Don't use Spurr's! } } } Bye, } hildy }
I would appreciate tips on preparing marine bacteriophages for TEM - a totally new departure for us. At present, they will have to be examined at ambient temperature.
Ted Dunn's suggestion would not put an end to commercial misuse on this listserver. It is NOW a requirement that vendors must write a disclaimer when mentioning products where a potential conflict of interest exists. Most of my contributions do not require a disclaimer. His suggestion was prompted by repeated misuse of the listserver by one person. 1 The offender has repeatedly used the disclaimer as an advertisement itself. 2 Similarly, it is against the rules for a vendor to proffer product information unless it's a direct reply to a question. The offender has made a habit of changing topics and uses other posting as a springboard for his own agenda. It's advertising on the cheap and rarely informative. 3 It is in poor taste and in deed offensive for a vendor to make general statements that "his" products are somehow superior to those from other vendors (nothing wrong in saying "our apertures have two holes", if a specific reason is given why that may be advantageous). The offender's postings alludes that his products are somehow, in a general way superior. 4 It is not furthering the aims of this listserver for a vendor to pretend to reply to a question, but then gives a misleading answer designed to steer towards the vendors products, rather then provide simple technical help. Trouble is that some readers look for information and do not have the experience to separate chaff and wheat; they stand to lose much time and some money.
Sure, occasionally we have a vendor run amok, making a single posting which is quite contrary to the rules with only a risk of ostracism. But why is this one offender allowed to persist after dozens of offending postings? I once asked him why he persists with these postings and the reply was: "It works, you should see the counter on my webside move after I've made a posting."
Dozens of suppliers read the listserver, almost all play by the rules most of the time. Precedents can cause problems: What if another and then many vendors decided to also abuse the listserver? Cutting them off could generate an interesting court case. The evidence is easy to find: It's all in the archives and the last major offence occurred in the recent "formvar" thread. My slightly acid reply to that posting was the first that has not resulted in some form of bullying, including threads to a common suppliers.
A bit of social science: Clever people are adept at finding reasons, right or wrong. Good judgement is a separate trait. Many of us also avoid confrontation or prefer to shoot the messenger. Surely the issue here is: What kind of listserver do we want? Should a person be allowed to make postings which with monotonous regularity offend as outlined in the above four points? Cheers? Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Wednesday, 14 October 1998 10:15, "DUNNTEM-at-aol.com"-at-sparc5.microscopy.com [SMTP:"DUNNTEM-at-aol.com"-at-sparc5.microscopy.com] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } I recently read the postings relating to a vendor using } this List in a } somewhat roundabout way to advertise! } } I have a proposal to make which I believe would put an end } to this. } } Vendors should all use the same simple disclaimer in their } posts. This would } read simply: "We are a vendor of electron microscope } supplies" or words to } that effect. } } In addition, the body of the posting should address itself } only to the } specific questions being answered and not slip in } statements like "...which } this company sells....etc" unless someone is asking } directly for a source. } } Any comments? } } } Ted Dunn } Maui, Hawaii }
Energy Beam Sciences has a position available for a Sales Executive/Product Specialist at our facility in Agawam, Massachusetts. Anyone interested in more details should contact me back-channel.
We have used KM4 and HM20 on at least a hundred occassions. We work in a standard fume hood and wear gloves. No allergic reactions have ever occured. Do you have some reason to think this is more toxic than the other resins?
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Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
This discussion is very interesting to me. I have been thinking about taking an old hard drive apart and trying to image [a piece of = the platter] the magnetic domains, particularly the format and data dots. = This sounds pretty ridiculous since the domains have no visual or elemental variations, but it was my intention to use something like iron powder = as a disclosing agent, Since static cling dominates (over gravity and = magnetic forces) in that particle size range, I was only going to expose the = piece to the airborne dust of iron powder [how"?]. This shouldn't be the size of magnetic forces that interfere with beam dynamics, so I think its = do-able. Anybody else tried something like this or thought about it?
Most of my work involves finding silicone or other organic contaminants on the surface of airplane parts. I use AUGER and ESCA, = but just wanted to get an SEM view of the platter.=20
} =20 } =20 } There are a number of articlea on magnetic domain imaging. The idea = is to } keep } the magnetic material away from the polepiece. As far as charging of = the } package } goes one could go to a low KV or paint the non-conductive package = surface } with } conductive paint. } =20 } Good Luck, } =20 } Earl Weltmer } =20 } P.S. Can we pick on Barbara some more? } =20 } BCarmic424-at-aol.com-at-sparc5.microscopy.com wrote: } =20 } } = ------------------------------------------------------------------------=
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of = America } } To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com } } On-Line Help = http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } = -----------------------------------------------------------------------.=
} } } } Dave, } } } } } From my point of view (as an EM service engineer), introducing a = magnet } into } } any area in an SEM can cause trouble. Even brief contact between a } magnetic } } sample and EM stages and sample exchangers can cause them to = become } slightly } } magnetic themselves. Even if your stage is made of brass or = aluminum, } there } } are still setscrews and other mechanisms which might need = degaussing if } the } } imaging of the scope is affected. } } } } Sincerely, } } } } Bill Carmichael } } } } Allequash Engineering } =20 } =20 } =20 } =20
{bigger} {bigger} MME {/bigger} {/bigger} is America's first national consortium dedicated to
customized on-site training in all areas of
microscopy, sample preparation, and image analysis.
At 08:29 AM 10/14/98 +0100, Colin Reid wrote:
} } } }
{excerpt} {smaller} Hi,
{/smaller}
{smaller} I would be interested in hearing from anyone who is working on remote control of EM's across the Internet. We have people who are interested in this and it would help greatly if we could get some pointers.
Your material will have to be cryo sectioned otherwise you will find too much smearing and deformation . We tend to section such materials at about -120 C. Diamond knives work better, but glass knives should do OK. You can stain the nylon with phosphotungstic acid and the B in ABS with osmium tetroxide. The PP will stain somewhat with ruthenium tetroxide.
I hope this helps.
Jordi. ---------- } From: "740206-at-ucl.itri.org.tw"-at-sparc5.microscopy.com To: Microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
It is not necessary to use propylene oxide as an intermediate agent, even if the embedding media is viscous (contains Araldite 502). One can use acetone with great success. It is important, however, to really "rinse out" all the acetone with resin. Remnants of acetone or alcohol, unlike propylene oxide, do not become part of the polymerized system, and will interfere with polymerization. We have not used propylene oxide in 7 years. There was one exception: Someone brought me very valuable blocks which had been badly embedded. I managed to pull out the bad embedment with PO over a three day period and reembed, section, treat it with radioactive label, etc. The micrographs were recently published. Otherwise we have not used PO. If you decide your epoxide embedding medium is too viscous, take advantage of the mechanical property of epoxides to become fluid between 37degC and 45deg. Put the vials on a rotator, and add a 60Watt light bulb to the setup. Position the lamp so that the mixture does not exceed 45 degrees. Wonderfully successfull for difficult or large specimen. Protect yourself! Get rid of Spurr's! Carcinogens are cumulative. Hildy
-----Original Message----- } From: HILDEGARD CROWLEY [mailto:hcrowley-at-du.edu] Sent: Wednesday, October 14, 1998 10:05 AM To: postmessage
Hi, Would somebody on this listserver be able to explain to me the "Balch procedure"? Suposedly this procedure permeabilize cells, and was introduced by William Balch. All correct answers will be appreciated. J. Gabrovsek CCF Cleveland, Ohio
There are three ways to image the magnetic domains directly in the SEM. = Two are readily available in any SEM. One uses the backscattered signal = and the other uses the secondary signal. Which one depends whether the domains = are oriented in the plane of the sample (BSE-I think) or perpendicular(SE - = I think). Take a look in the SEM book by Goldstein et al. and I think = that the type I and type II imaging modes are described there.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Chavez, Tim A To: 'MSA Miscroscopy Listserver' = -----------------------------------------------------------------------.=
This discussion is very interesting to me. I have been thinking about taking an old hard drive apart and trying to image [a piece of = the platter] the magnetic domains, particularly the format and data dots. = This sounds pretty ridiculous since the domains have no visual or elemental variations, but it was my intention to use something like iron powder = as a disclosing agent, Since static cling dominates (over gravity and = magnetic forces) in that particle size range, I was only going to expose the = piece to the airborne dust of iron powder [how"?]. This shouldn't be the size of magnetic forces that interfere with beam dynamics, so I think its = do-able. Anybody else tried something like this or thought about it?
Most of my work involves finding silicone or other organic contaminants on the surface of airplane parts. I use AUGER and ESCA, = but just wanted to get an SEM view of the platter.
} } } There are a number of articlea on magnetic domain imaging. The idea = is to } keep } the magnetic material away from the polepiece. As far as charging of = the } package } goes one could go to a low KV or paint the non-conductive package = surface } with } conductive paint. } } Good Luck, } } Earl Weltmer } } P.S. Can we pick on Barbara some more? } } BCarmic424-at-aol.com-at-sparc5.microscopy.com wrote: } } } = ------------------------------------------------------------------------=
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of = America } } To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com } } On-Line Help = http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } = -----------------------------------------------------------------------.=
} } } } Dave, } } } } } From my point of view (as an EM service engineer), introducing a = magnet } into } } any area in an SEM can cause trouble. Even brief contact between a } magnetic } } sample and EM stages and sample exchangers can cause them to = become } slightly } } magnetic themselves. Even if your stage is made of brass or = aluminum, } there } } are still setscrews and other mechanisms which might need = degaussing if } the } } imaging of the scope is affected. } } } } Sincerely, } } } } Bill Carmichael } } } } Allequash Engineering } } } }
Many years ago I abondoned formvar films for butvar films on grids. Butvar is "stickier", it is incredibly stable so that very thin films can be used. Butvar withstands repeated entry into the microscope. The films seem to last for years in storage dishes. Butvar B-98 (available from Electron Microscopy Sciences both in powder and liquid form)(I, unlike other microscopists have not amassed great wealth, and therefore own no stocks in EMS), also at high magnification displays a lot less "texture". I check every new film batch the following way: All butvar films (this is in published reports) display at very high magnifications very small isolated pinholes. I try to find one of these and turn the beam at crossover on the hole. Nothing happens - no stretching, no tearing, no movement. Butvar is used like formvar, except that it is dissolved in chloroform. It is floated off slides, etc, picked up on clean grids, etc. It requires clean glassware and a quiet casting environment. The 0.25% solution from EMS is useful for 25-50 mesh grids. Perfectly stable under most conditions. For large slot grids, it is best to use the powder at a concentration of about 0.75%. The film is so electron lucent that one can easily increase the concentration without excessive loss in contrast. Dissolving the powder can be done with heat, or by standing the bottle in the hood for about 3 days with occasional swirling. For use of the powder, please look up the reference below. Handley, D., Ultramicrotomy, 4 (1979) 479-480. Another useful feature of the butvar is that it is very sticky. If one dilutes it to 0.15% and places a drop on each clean grid lying on 2 layers of filter paper, the grids hang on tenaciously to epoxy sections. No loss during staining, etc. The "vest" of butvar is not detectable in the scope. For 10 years I have avoided the certain insanity caused by section loss during poststaining of grids by simply "vesting" all grids in butvar. I have not tried to do this with formvar.. It probably works well also. If there is section loss during protracted manipulation of grids, then the grids are warmed overnight in a 50 deg oven on the filmed grids. Presto. Glued together. (Do not flood grids while vesting. You will not be able to get them off the filter paper). I have not tried other films. There may be something even better out there. Have fun! Hildy
A back of an envelope calculation with my after dinner coffee comes up with the following numbers Radius of the Earth 6379 km Radius of a US quarter 12.25mm
Magnification comes out 520.73469 x 10 to the power 6
If Alvin Eades is worrying about wear along the milled edge of the quarter; where do we measure the earth's radius ? From the top of Everest +8848m or the bottom of the Dead Sea -394m ?
I'll settle for 520 million.
Patrick Echlin Cambridge
On Wed, 14 Oct 1998, Mark Darus wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Earth Diameter = 12,756.28 KM -at- Equater } Quarter Diameter (US) = 7/8 inch } } The Earth is 573,960,854.9 times larger, so that's your mag. } }
Fryer Co. (Huntley,IL) has openings for a Marketing Manager and Sales Personnel for our family of specialized biomedical research microscopy products.Specifically the product line consists of modules for light microscopes (any brand) which can perform the following biological investigations: Nearfield microsocpy (NeD); UV photolysis through a fiber optic probe; quantitative fluorescence; emission ratioing using dual cameras or high speed photomultipliers; detection of selected regions of fluorescence emission integrated with electrophysiology recordings. Mktg Mgr is based in Huntley, IL (near Chicago) while sales positions are located in key geographic locations in the U.S. Fryer Co. has been a NIKON microscope dealer for over 30 years and additional product lines include: Universal Imaging Corp, Media Cybernetics, Sony, Roper Scientific, etc. Positions report to VP-Mktg. Send resume to Roger Main via e-mail at roger-at-fryerco.com, fax to 847-669-2056, or mail to 11177 Dundee Rd, Hunt ley, IL 60142. EOE
} } Fonts/Colors etc are fine but they do not sure a sufficiently useful purpose } in Email. They just cause headaches for some subscribers. These items } belong on a WWW page and NOT imbedded in an Email message..... } }
The bottom line is that lots of folk simply will not read email that necessitates separate downloading and viewing. If folk want their email to get read, they need to send it in a format that folk will read. If folk don't want their email read, then why send it?
I have been doing negative staining on bacteriaphage from Lactobacillus. As milk is a highly osmotic meduim and my technique works well on this phage, it may work well with marine-based phage.
Negative stain with 200mM Uranyl oxalate, pH 6.8 Make stain immediately prior to use by adding equal amounts of 200mM uranyl acetate and 200mM oxalic acid. Adjust pH with 200mM ammonium hydroxide. Adsorb virus to carbon coated grid for 2 to 4 minutes. Do not rinse. Put grid on drop of UO stain for 2 min., wick off excess stain and let dry (~1hour). With this technique I am able to resolve the the individual proteins in the phage tail.
Bill
} Dear List } } I would appreciate tips on preparing marine bacteriophages for TEM - a } totally new departure for us. At present, they will have to be examined } at ambient temperature. } } Keith Ryan } Plymouth Marine Lab., UK } } PS - Hello, Danielle! } }
William R.McManus Supervisor Electron Microscope Facility Department of Biology Utah State University Logan UT 84322-5305
I agree with Alwyn Eades completely. This individual would not have posted the request if they had the means to perform the calculation themselves. BTW, my US quarter measured 0.952 inch (with calipers), so the mag is really 528 million.
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Although I prefer using Epon generic resins for many tissues, we use Spurr's regularily for plant tissue which requires a very low viscosity resin and careful, very slow infiltration to penetrate the heavy cell walls and give the best results. We get excellent staining for both LM and TEM.
We polymerize at 60oC for 48hrs. and use the standard Spurr's formulation. We also use gloves and work in fume hoods to minimize health hazards. The oven used for polymerization also sits in a fume hood.
Staining for TEM usually consists of 2% UA for about 5 min (this can be eliminated if you do en block staining prior to dehydration), followed by Reynold's lead citrate (original formulation) for 3 min in an NaOH atmosphere.
We usually use 1% aqueous toluidine blue for staining thick sections. We put our slides on a hot plate set at a very low temperature to stretch our sections in small droplets of water and then adhere them to the slide. Staining is then also carried out on the hot plate. Staining is much faster and more effective if you cover your sections with the Tol. blue and then add a drop or two of 1% aqueous Na Borate to raise the pH. You can mix the toluidine blue and Na borate together but it is not a very stable solution. Both will keep for months with no problems if kept separate.
I have stained larger numbers of slides by putting them in a coplin jar filled with the toluidine blue-Na borate solution (~1:1) diluted down to about 1/10 the original concentration. The coplin jar is then put into an oven (~60oC) for a prolonged period of time. The timing depends on the tissue and may be for 15 min or even 1 or more hours. Once you have it figured out for your material, you can efficiently batch stain.
There are some other low viscosity resins on the market but I have not had experience working with them. Perhaps there is one that is safer to use than Spurr's and will give equal or better results. Let's hear from other list members about alternatives.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Hi,
Spurr's is the most highly crosslinked of all embedding media used for LM-TEM. The higher the crosslinkage, the less likely any stain, whether for LM or heavy metal for TEM, is to penetrate well enough to be called successfull. First: Get rid of Spurr's. It contains a potent carcinogen, VCD. Do you need this? Second: If you cannot get rid of Spurr's (try real hard), then use the softest mixture you can tolerate for cutting. Polymerize at 60deg C overnight, and see if this is adequate. This will reduce the final crosslinkage. Third: Blocks in existence already! Don't stain well? Try soaking the sections prior to staining for extended periods of time in water, then 5 min in alcohol, in increasing concentrations until you reach 95%. Then go back down to water stepwise. Try staining. Fourth: Use as alkaline as possible a vehicle for your stain. pH 12 is about right. Fifth: Combine all of the above. Does not work? Soak the sections in water and gradually bring to 95% alcohol.. Expose to stain dissolved in alcohol. Does not work? Forget it. Start over.
If you have very valuable sections and you must stain them for TEM, use alcoholic UA for 10min at 60deg C. Use Reynolds lead citrate at a pH of about 9 or 10. This last trick is truly a last resort, since the lead may dump erratically (or stain easily) at this low pH.
If you polymerize a block at a low power for 45 minutes in a microwave, you crosslink the resin to such an extent that nothing, nothing, nothing you do will stain it. (Lost my best meat loaf dish this way). For TEM we do not polymerize resins totally, about 10% of the monomers are left unreacted with one another. If you "drive" the crosslinkage, the block will be harder, less elastic, and impenetrable for liquids (except if you boil it for a year or so in water). I am not exaggerating. I got this info out of one my very favorite materials science books.
Don't use Spurr's!
Bye, hildy
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Michael, If you intend to use your tissue for immunocytochemistry, keep the polymerization temperature below 60o to protect the antigenic sites from being denatured. We usually keep the temperature around 55o and polymerize= in a nitrogen atmosphere. Alternative is the cap your capsules in such a way to exclude as much air as possible.
You do have to use precautions with the lowicryls. It is a good idea to double glove when handling them and work in a fume hood. If LRWhite works Ok then stick with it as it is about the safest resin you can use....however, if you do not get sufficient antigenicity, then lowicryls m= ay be worth using and just take the necessary precautions to not get it on your skin.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University =20 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Dear collegues, =09=09thank you for all the interesting and diversificated hints for my LR-White embedding procedure. Yesterday I started embedding samples in LR-W= hite without the accelerator/catalyst, both in the oven at 60=B0C as well as at -20=B0C in the UV-fridge in order to compare the grade of ultrastructural resolution. I=B4ll tell you about my findings anyhow ...
Finally I=B4d like to ask about the possibly risks of Lowicryl because Debby Sherman proposed to try out Lowicryl HM20. A few months ago we ordere= d this stuff but up until now nobody agreed in working with this substance fearing possible risks on health and life so that the delivery box remains not opened somewhere in our Lab ... :-( Anyone, especially Debby, could tell me about experiences in working with Lowicryl (HM20/K4) and the REAL risk of allergies? Are there secure working conditions to avoid hazard? I=B4d like to know whether it=B4s worth= trying out this or if it=B4s better to keep hands off.
With best regards, Michael
Michael Reiner Department of Anatomy University of Cologne (Germany) Joseph-Stelzmann-Str.9 50931 Cologne (K=F6ln)
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On Thu, 15 Oct 1998, William R. Oliver wrote: } The bottom line is that lots of folk simply will not read } email that necessitates separate downloading and viewing. } If folk want their email to get read, they need to send } it in a format that folk will read. If folk don't want their } email read, then why send it?
Regarding recent posts by J. Darley and T. Dunn: I use several kinds of listservers and newsgroups, and I'm not a vendor. Vendors' comments are very often quite helpful to me. I don't think I've ever had a problem with a vendor trying to "foist off" his product line on me. Almost always they are proud of their business and clearly identify themselves and their business address.
It seems to me vendors citing their products are little different than scientists trying to get you to use their method or read their publications. Let's encourage vendors to contribute technically, as long as they don't post their entire product line, or automatically link you to their Home page. After all, if vendors are not part of our conversation, how will we get them to make the products we need?
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I have used Lowicryl resins for many years with excellent results. = However, the Lowicryl resins are (meth-)acrylic acid ester preparations = and the toxicological properties of these products is not fully known. It = is clear that (meth-)acrylic acids can cause allergies. This is important = to bear in mind when handling these resins.
Here are some tips to safe handling: *Work with them in a well ventilated area. *Use protective clothing and eye protectors. *Use protective skin creams and appropriate gloves. Latex gloves are not = fully protective (the resins, once on the gloves, can penetrate in a few = minutes!). 4H gloves are recommended. *Avoid direct skin contact from liquid or vapour (remember the vapours = also cause allergic reactions).
I have met researchers who develop no reaction to the resin (myself = included) while others develop serious skin reactions, instant headache or = sudden nausea when in contact with the vapour. Once sensitized, there is = no protection. Gloves and respirators offer no relief.
Having said all that, I use Lowicryls in a chemical hood, but without = gloves! Having seen the way gloves are misused in the laboratory, I feel = that for some special instances, no gloves and lots of care is best. I = say this because gloved workers tend to feel a false sense of total = protection. Gloves stay on when moving around the laboratory and large = areas of bench become contaminated with whatever was spilled onto the = glove. =
Without gloves, that rare spill is immediately detected and washed off = instantly. With gloves, the spill can often work through and remain in = contact with the skin for some time before being washed off.
I am sure there will be lots of negative comments about this, but bear in = mind I did say " for some special instances". I certainly wouldn't teach = this method to students or new arrivals into the lab. It is an educated = risk I take.
Did anyone try out the MonoStep resins, which were introduced to replace = the Lowicryls?
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org
I have problem trying to follow up on DiI labeled cells engrafted in mouse heart. When trying to stain them with whatever ab, the DiI diffuses/dissapears. Any suggestions on how to make it stay, so I can see where my cells are? The sections are 10 microm thick, frozen.
Thanks, Catalin Toma Johns Hopkins University School of Medicine Division of Cardiology Ross 812 Baltimore, MD 21205
I have been notified that after much delay the following conference proceedings are now available. Although they are obviously for sale, I believe that a short, one time, announcement is a reasonable courtesy.
Nestor Your Friendly Neighborhood SysOp...
------------------------------------------------------- Appended Text Edited by NJZ -------------------------------------------------------
Proceedings of the 14th Pfefferkorn Conference The Science of Biological Specimen Preparation for Microscopy Scanning Microscopy Supplement 10, 1996 ISSN: 0892-953X / ISBN: 0-931288-49-5 Edited by Marek Malecki and Godfried M. Roomans
Proceedings of the Thirteenth Pfefferkorn Conference Scanning Microscopy Supplement 9, 1995: Luminescence ISSN: 0892-953X / ISBN: 0-931288-48-7 Edited By: G. R=E9mond, L. Balk, and D.J. Marshall
Additional Details can be obtained directly from the publisher.
Scanning Microscopy Intl., Box 66507, AMF O'Hare (Chicago), IL 60666-0507, USA =46AX: (847) 985-6698 / E.mail: 73211.647-at-compuserve.com
I am in urgent need to get as cheap as possible (generally counting for one of the "FOC or ..take away for transport costs" solutions) the HT units form Philips EM301 TEM microscope. Preferably from Europe. We try to help one of our customers by equipment officially ,,out of service". We need specially A-units, X-unit, Z-units, eventually the whole HT-TANK. We have strange problem with some interferences in HT circuit so that when the modules are removed out from microscope on extension cables work sometimes O.K. but when installed back or relocated on desk do not regulate HT and it's going to maximum or even higher - this is known from the image - maximally overfocused intensity covers only half of the screen by highest lens settings.
appreciate any help
Krzysztof Herman FEI/Philips EO Service - Poland Labsoft, ul.Bazancia 45 A 02-892 Warszawa tel/fax:(+48 22)6446233 E-mail: kherman-at-labsoft.com.pl http://www.labsoft.com.pl
You'll find that the magnetic domains are quite visible directly on SEM examination. 16 years ago, I helped design an SEM for IBM that would image any portion of a 14" hard drive disk (very large sample chamber). Their primary concern was to image the magnetic material grain size and distribution, but it also enabled them to read the data and measure the magnetic domain distribution.
The magnetic fields show up as brightness variations that are the result of the different magnetic polarities having a differing effect on the production of detectable secondary and backscatter electrons. Depending on the fields present, the effect can be subtle. Two methods have been recognized - the first and simplest, is detection by secondary electrons. The secondary electrons emitted have their trajectories affected by the magnetic fields. Resolution of individual domains is not as good as the second method but imaging is easier.
The second method uses the detection of backscatter electrons. When the magnetic domains have their field axis parallel to the sample surface and perpendicular to the beam, a large beam to sample angle will produce a differential production of backscatter electrons because of the cyclotron effect, where the electrons will tend to move one way or another around the field, depending on the polarity. Those electrons moving closer to the surface will produce more backscatter electrons.
This method, while producing greater domain resolution, also produces a very low contrast (i.e. - crank up the gain).
Larger magnetic fields will affect the beam position also, giving very strange images, although you won't see this affect from magnetic recording media.
} This discussion is very interesting to me. I have been thinking } about taking an old hard drive apart and trying to image [a piece of } the platter] the magnetic domains, particularly the format and data } dots. This sounds pretty ridiculous since the domains have no visual } or elemental variations, but it was my intention to use something } like iron powder as a disclosing agent, Since static cling dominates } (over gravity and magnetic forces) in that particle size range, I } was only going to expose the piece to the airborne dust of iron } powder [how"?]. This shouldn't be the size of magnetic forces that } interfere with beam dynamics, so I think its do-able. Anybody else } tried something like this or thought about it? } } Most of my work involves finding silicone or other organic } contaminants on the surface of airplane parts. I use AUGER and ESCA, } but just wanted to get an SEM view of the platter. } } } Tim Chavez } 316-526-5394 desk } 316-526-1851 fax } } tim.chavez-at-wichita.boeing.com } } } __|__ } _______O_______ } =B0 =B0 } } ________________________ } Dyslexics of the world untie! } } } } } } ------------------------------------------------------------------ } } -----. } } } } } } There are a number of articlea on magnetic domain imaging. The } } idea is to keep the magnetic material away from the polepiece. As } } far as charging of the package goes one could go to a low KV or } } paint the non-conductive package surface with conductive paint. } } } } Good Luck, } } } } Earl Weltmer } } } } P.S. Can we pick on Barbara some more? } } } } BCarmic424-at-aol.com-at-sparc5.microscopy.com wrote: } } } } } ---------------------------------------------------------------- } } } -------- The Microscopy ListServer -- Sponsor: The Microscopy } } } Society of America To Subscribe/Unsubscribe -- Send Email to } } } ListServer-at-MSA.Microscopy.Com On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------- } } } -------. } } } } } } Dave, } } } } } } } From my point of view (as an EM service engineer), introducing } } } } a magnet } } into } } } any area in an SEM can cause trouble. Even brief contact } } } between a } } magnetic } } } sample and EM stages and sample exchangers can cause them to } } } become } } slightly } } } magnetic themselves. Even if your stage is made of brass or } } } aluminum, } } there } } } are still setscrews and other mechanisms which might need } } } degaussing if } } the } } } imaging of the scope is affected. } } } } } } Sincerely, } } } } } } Bill Carmichael } } } } } } Allequash Engineering } } } } } } } } } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Let me inform you that the 'Exhibition' link (http://www.eurem2000.isibrno.cz/exhib.html) was added to the website of the '12th EUROPEAN CONGRESS ON ELECTRON MICROSCOPY'.
Producers of microscopes and related instrumentation, of laboratory equipment, materials and tools for microscopy, and publishers of scientific literature are asked to fill in the Questionnaire for Exhibitors .
Best regards,
Petr Schauer
+---------------------------------------------------------------------+ | Dr. Petr Schauer, Vicechairman of the Or- | tel.: (+420 5) 41514313 | | ganization Committee of the 12th EUROPEAN | fax : (+420 5) 41514404 | | CONGRESS ON ELECTRON MICROSCOPY | (+420 5) 41514337 | | (Brno, Czech Republic, July 9 - 14, 2000) | e-mail: petr-at-isibrno.cz | | Kralovopolska 147, CZ-612 64 Brno | eurem2000-at-isibrno.cz | | Czech Republic |www.eurem2000.isibrno.cz | +---------------------------------------------------------------------+
Some references for the "Balch procedure" are listed below. See 1989 for early description, and 1996-97 papers for references to numerous 1994 and other citations.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Balch WE, Mccaffery JM, Plutner H, Farquhar MG. 1994. Vesicular stomatitis virus glycoprotein is sorted and concentrated during export from the endoplasmic reticulum. Cell 76(5 11C):841-852.
Bannykh SI, Rowe T, Balch WE. 1996. The organization of endoplasmic reticulum export complexes. J Cell Biol 135(1):19-35.
Beckers CJM, Keller DS, Balch WE. 1989. Preparation of semiintact Chinese hamster ovary cells for reconstitution of endoplasmic reticulum-to-Golgi transport in a cell-free system. Meth Cell Biol 31:91-102.
Nishimura N, Balch WE. 1997. A di-acidic signal required for selective export from the endoplasmic reticulum. Science 277(5325):556-558.
Rowe T, Aridor M, McCaffery JM, Plutner H, Nuoffer C, Balch WE. 1996. COPII vesicles derived from mammalian endoplasmic reticulum microsomes recruit COPI. J Cell Biol 135(4):895-911.
Tisdale EJ, Balch WE. 1996. Rab2 is essential for the maturation of pre-Golgi intermediates. J Biol Chem 271(46):29372-29379.
} Hi, } Would somebody on this listserver be able to explain to me } the "Balch procedure"? } Suposedly this procedure permeabilize cells, and was introduced by } William Balch. } All correct answers will be appreciated. } J. Gabrovsek } CCF Cleveland, Ohio }
Email: reznor-at-holly.colostate.edu Name: Glen El-Hayek School: Colorado State University
I was wondering if there was anywhere I could find info on what exactly freeze fracture electron microscopy is and how it works. I am working on a web site for a cell bio class and that is the topic we have to research.
Thank you very much, Glen El-Hayek Colorado State University
A co-worker said he could not stain his sections on Butvar coated grids with alcoholic UA. The film broke everytime. But if he used aqueous UA he had no problem. Have you experienced any problems staining Butvar with alcoholic UA?
George Lawton Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas
I also have a Sony printer (UP-D8800, 8.5"x11" dye sublimation) and experience problems with permanence. Initial output looks good, but after only a couple of weeks on the desk (no sunlight, just standard fluorescent lighting) prints are noticably faded. A simple sheet protector seems to keep that from happening, but it is annoying to have to put everything in one. Print cost for the UP-D8800 is about $2 per color print. I mentioned this to Sony at the MSA meeting in Atlanta. Their only suggestion was to use a different print paper, that accepts a laminated cover film. The cost and description of the paper made me decide to stay with sheet protectors, at least for now.
Interestingly, we have a smaller format Sony (UP-1800MD, as I recall; prints on paper ~4"x5.5") that does not have this problem. I guess they are using different print systems (Sony--hint?)
I haven't noticed any transfer of "ink" for either of these printers.
Jim Passmore Analytical Chemist Cryovac Division, Sealed Air Corporation P.O. Box 464 Duncan, SC 29334
---------- } From: corwinl } To: Microscopy } Subject: Prints: permanence } Date: Friday, October 09, 1998 12:04PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I agree with you about not using propylene oxide but my concern about acetone is that it has a habit of getting through gloves quickly or rendering them more fragile or porous so that you are more likely to be exposed to a less carcinogenic resin. I still feel safer about using Spurr's as a universal resin and LR White as a safe alternative, when possible. It means that we hold less types of epoxide resins and everyone is taught to treat Spurr's with great respect. We always wear gloves and try never to bring the gloves outside of the fume hood unless polymerized. All non-polymerized resin operations are ALWAYS carried out inside our fume hood including polymerization and I have tried to eradicate any sources of resin dust such as sawing of blocks, unless again done in the fume hood. I do, however, take your point about Spurr's and will continue to ponder it.
It would be great if we could completely remove the need to use cacodylate, osmium, propylene oxide, Spurr's. lead and uranium (not to mention glutaraldehyde, formaldehyde, lowicryl, ruthenium, tungsten and molybdenum) but the alternatives don't always seem to work or are just more difficult to manage in other ways. Perhaps we should all get robots or tissue processors.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
Disclaimer - My opinions again etc
---------- } From: HILDEGARD CROWLEY To: postmessage
Hi,
It is not necessary to use propylene oxide as an intermediate agent, even if the embedding media is viscous (contains Araldite 502). One can use acetone with great success. It is important, however, to really "rinse out" all the acetone with resin. Remnants of acetone or alcohol, unlike propylene oxide, do not become part of the polymerized system, and will interfere with polymerization. We have not used propylene oxide in 7 years. There was one exception: Someone brought me very valuable blocks which had been badly embedded. I managed to pull out the bad embedment with PO over a three day period and reembed, section, treat it with radioactive label, etc. The micrographs were recently published. Otherwise we have not used PO. If you decide your epoxide embedding medium is too viscous, take advantage of the mechanical property of epoxides to become fluid between 37degC and 45deg. Put the vials on a rotator, and add a 60Watt light bulb to the setup. Position the lamp so that the mixture does not exceed 45 degrees. Wonderfully successfull for difficult or large specimen. Protect yourself! Get rid of Spurr's! Carcinogens are cumulative. Hildy
by srvr22.engin.umich.edu (8.8.8/8.8.8) with ESMTP id LAA27914 for {microscopy-at-msa.microscopy.com} ; Fri, 16 Oct 1998 11:15:26 -0400 (EDT) X-Sender: bigelow-at-srvr5.engin.umich.edu Message-Id: {v03007800b24d128022df-at-[141.212.131.74]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I must say that I don't see what all the fuss is about. Most vendor's comments that I have seen on this listserver have been in response to a question raised by someone else, and have, in my opinion, been technically oriented and highly helpful. Several of the vendors who participate in this listserver also operate consulting laboratories, and thus have lots of experience in methods and techniques. Also, one of the big problems in our line of work often involves finding a material or a piece of apparatus to meet some particular need. If a vendor has such an item readily available, I think it is very helpful to know about it.
I say, unless things really get much worse, let's just relax and delete any messages we don't want to read.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
I think you will find the Freeze-Fracture method discussed in some detail in the book 'Low Temperature Methods in Biological Electron Microscopy', by Robards & Sleytr, Elsevier, 1985 (ISBN 0-444-80684-9 or -7) which is Vol 10 in the series 'Practical Methods in Electron Microscopy' that is edited by Audrey M. Glauert
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
As far as I can see, those vendors who contribute regularly to the list offer helpful hints from their perspective, state their commercial interest, and usually state if they are not the sole supplier of product XXX. The only occassional problems I have noticed have been with vendors who are not regular contributors coming out of the blue with blatant advertising (usually follwed by several flames from annoyed subscribers).
Why don't we leave the vendors alone for now and leave this discussion behind. They have something useful to contribute to this discussion group (and most of the rest of us couldn't work without them anyway). Also, Nestor is the person to contact if you have problems with a specific mail.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, or: ianmaclaren-at-hotmail.com Birmingham B15 2TT, http://web.bham.ac.uk/I.MacLaren England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
There have been several reports of butvar being dissolved in alcohols. I have no recollection of using alcohol on butvar films, but I believe that those who have volunteered th information are correct. Caution. Try it first. Also, butvar is a lot more hydrophilic than formvar. I used to love it for negative staining. Further (I cannot explain this) I had wrinkles in LR White while using someone's formvar coated grids. When using my butvar grids the wrinkles did not occur. The butvar polymer is known for its flexibility. Perhaps it is able to "adjust" better to section size. I don't know. I have several messages regarding butvar from people when they used the "Reply and include original message in the reply" function. I did not get the messages. There must have been at least 5 of these. If you need more information, ask me again, but do not send back my original message. I have been having all sorts of strange troubles with the University System. Bye, Hildy
Ken Converse said: } Chuck Garber and others identify themselves quite clearly and I have no } problem whatsoever with his, or others' postings. What really grates on } my nerves is people who think there is something wrong with being a } business person. Not everyone can run away and hide in an ivory tower } and not everyone is suited to being someone else's employee. Give us } business owners a little slack. Chuck offers a lot of good information } and sources besides SPI. I think that some subscribers could even offer } him an apology for their rude postings. } } Ken Converse } owner } Quality Images } Delta, PA } third party SEM service } } Opinions expressed are mine. Yes, I sell SEM services for money. No, I } do not apologize for having opinions or charging for my SEM services so } that I can earn a living and my customers' SEMs will run well.
As a manager of a University EM facility, I can use all the help I can get in regards to new products and methods availble. I have felt that vendors are a resource not an anoyance. Now, the people that call my home six times a night selling refinancing and long distance services, now they are annoying.
Bill
William R.McManus Supervisor Electron Microscope Facility Department of Biology Utah State University Logan UT 84322-5305
I have been using epon (and very successfully, I might add) for over 15 years without benefit of PO as an intermediate. My original procedure used graded acetone (100% was stored at -20C over molecular sieve) but I switched to EtOH without any negative impact. Despite the older references which state that EtOH is not miscible with epon, I find that miscibility with the replacements for Epon 812 is very good. The procedure requires that good mixing of the epon and EtOH is thorough!! After 3 changes of 100% EtOH, the tissue is processed through graded EtOH/epon (2:1, 1:1, 1:2), into pure epon (the mixture, of course) for three changes, the last two being 4 to 8 hours each (of course, it also helps to have an EM tissue processor to permit overnight processing!).
I abandoned Spurr's because of the staining problems, the toxicity, an acquired dermal allergy and the hardness of the plastic. Following that acquired sensitivity, I then developed a similar contact response to PO. Even now, I double glove for epon, since there seems to be a cascade of allergic sensitivities that can develop over a lifetime of "familiarity breeding carelessness".
Hope this is of some help.
-----Original Message----- From: Jim J Darley [SMTP:jim-at-proscitech.com.au] Sent: Thursday, October 15, 1998 2:02 AM To: 'HILDEGARD CROWLEY' Cc: MSA.Microscopy.Com Subject: RE: Staining Spurr's???
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
Yes but, Spurr's has very low viscosity and does not require propylene oxide as an intermediate solvent. PO is a very powerful carcinogen. Most people seem to regard PO as essential with other epoxy resins. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Thursday, 15 October 1998 7:07, HILDEGARD CROWLEY [SMTP:hcrowley-at-du.edu] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } Hi, } } Spurr's is the most highly crosslinked of all embedding } media used for } LM-TEM. The higher the crosslinkage, the less likely any } stain, whether } for LM or heavy metal for TEM, is to penetrate well enough } to be called } successfull. } First: Get rid of Spurr's. It contains a potent } carcinogen, VCD. Do you } need this? } Second: If you cannot get rid of Spurr's (try real hard), } then use the } softest mixture you can tolerate for cutting. Polymerize } at 60deg C } overnight, and see if this is adequate. This will reduce } the final } crosslinkage. } Third: Blocks in existence already! Don't stain well? } Try soaking the } sections prior to staining for extended periods of time in } water, then 5 } min in alcohol, in increasing concentrations until you } reach 95%. Then go } back down to water stepwise. Try staining. } Fourth: Use as alkaline as possible a vehicle for your } stain. pH 12 is } about right. } Fifth: Combine all of the above. Does not work? Soak } the sections in } water and gradually bring to 95% alcohol.. Expose to } stain dissolved in } alcohol. Does not work? Forget it. Start over. } } If you have very valuable sections and you must stain them } for TEM, use } alcoholic UA for 10min at 60deg C. Use Reynolds lead } citrate at a pH of } about 9 or 10. This last trick is truly a last resort, } since the lead may } dump erratically (or stain easily) at this low pH. } } If you polymerize a block at a low power for 45 minutes in } a microwave, } you crosslink the resin to such an extent that nothing, } nothing, nothing } you do will stain it. (Lost my best meat loaf dish this } way). For TEM we } do not polymerize resins totally, about 10% of the } monomers are left } unreacted with one another. If you "drive" the } crosslinkage, the block } will be harder, less elastic, and impenetrable for liquids } (except if you } boil it for a year or so in water). I am not } exaggerating. I got this } info out of one my very favorite materials science books. } } Don't use Spurr's! } } } Bye, } hildy }
In a message dated 98-10-16 14:13:25 EDT, Professor Bigelow wrote:
{ { .....must say that I don't see what all the fuss is about. Most vendor's comments that I have seen on this listserver have been in response to a question raised by someone else, and have, in my opinion, been technically oriented and highly helpful. Several of the vendors who participate in this listserver also operate consulting laboratories, and thus have lots of experience in methods and techniques............. } }
As someone who is involved in the manufacture of electron microscopy supplies I agree very much with this sentiment. It is very valuable to have free exchange of information between scientists and those who produce the equipment and materials they require.
The point is though that there are inherent agreements on using this list. These agreements include low commercial profile by vendors. If one vendor frequently ignores that agreement and is allowed to do so then there is no question about it, he has a distinct commercial advantage. That is indisputable.
So the bottom line is, can we relax this rule and just leave everyone to post as they see fit or do we (vendors) all abide by the agreement.
Personally I would prefer the former since, as Professor Bigelow points out, one can always hit the delete button if not interested in a lengthy posting.
One great advantage of relaxing the rule would be that we didn't have to talk about it any more :-)
I have posted to this listserve before about the Sony printers. I have a UP-D7000 and it is basically a doorstop at this point. Both color and B&W prints fade within two weeks, or turn shockingly pink. This has been a problem for over three years now and I doubt that Sony has addressed the point. Their sales people tried to get us to buy one of their "latest and greatest" printers. Needless to say, their "L&G" fades just the same. Interestingly, I could substitute Phaser paper for the B&W and had no problems with output, even 3 years afterwards. Sadly, color prints didn't work with the Phaser paper.
If you want high quality dye-sub prints, my suggestion is to try the Kodak line, or the Codonics (using the same print engine). The prints cost more ($2.50/sh), but then you get what you pay for...
Cheers
-------------------------- John Bonevich NIST, Metallurgy Div. B164 Gaithersburg, MD 20899 USA TEL: (301) 975-5428 FAX: (301) 975-4553
} } I also have a Sony printer (UP-D8800, 8.5"x11" dye sublimation) and } experience problems with permanence. Initial output looks good, } but after only a couple of weeks on the desk (no sunlight, just } standard fluorescent lighting) prints are noticably faded. A simple } sheet protector seems to keep that from happening, but it is } annoying to have to put everything in one. Print cost for the } UP-D8800 is about $2 per color print. I mentioned this to Sony at } the MSA meeting in Atlanta. Their only suggestion was to use a } different print paper, that accepts a laminated cover film. The cost } and description of the paper made me decide to stay with sheet } protectors, at least for now. } } Interestingly, we have a smaller format Sony (UP-1800MD, as I } recall; prints on paper ~4"x5.5") that does not have this problem. } I guess they are using different print systems (Sony--hint?) } } I haven't noticed any transfer of "ink" for either of these printers. } } Jim Passmore } Analytical Chemist } Cryovac Division, Sealed Air Corporation } P.O. Box 464 } Duncan, SC 29334 } } ---------- } } From: corwinl } } To: Microscopy } } Subject: Prints: permanence } } Date: Friday, October 09, 1998 12:04PM } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for a supplier of Aquembed. Aparently it is some kind of resin that displaces water before cells are embedded in a material like Epon 812. I can't find Aquembed in the catalogs I have.
Does anybody know of a source (probably a private research lab working with primates) where I can find an anti-human IgG1 antibody that does not cross-react with monkey? (if I REALLY wanted to push my luck, I'd also stipulate that it is tagged with either rhodamine or fluorescein) I realize this is probably an impossible endeavor, but I have to exhaust all of my sources (Linscott's directory is not useful in this case). I would appreciate any help in this matter, Thank you, Laura
Laura M. Patrone, Ph.D. Wyeth-Ayerst Research BioMedical Imaging 641 Ridge Road Chazy, NY 12921 (518) 846-6318 patronL-at-war.wyeth.com
We have a Sony UP-D8800A (L&G) in one of my colleague's lab. There are several types of papers for this printer. The normal color paper does fade somewhat so we purchased the laminating type of paper and color rolls. I've printed some files for a poster on this and they came out very nice and they are still lasting and this has been at least 7 months. However, recently, I used it again and any areas of black and white on color prints are turning pink in about a week. I printed some other images on the B&W paper, and they still look good after several weeks. I can't say if it is because the paper and color rolls had been opened for some time or whether it was just a bad box. We have a new box and I will probably give it a try.
I have some prints that were done on a Kodak sub-dye about two years ago and they still look good. However, they haven't been exposed to a lot of light.
My HP 890C inkjet printer using the HP/Kodak Photo Deluxe paper is becoming my printer of choice for good quality B&W and color prints. It just about matches the Sony sub-dye (when the prints don't fade) and beats our old Seiko sub-dye every time.
-Scott Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: John Bonevich To: microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------.
I have posted to this listserve before about the Sony printers. I have a UP-D7000 and it is basically a doorstop at this point. Both color and B&W prints fade within two weeks, or turn shockingly pink. This has been a problem for over three years now and I doubt that Sony has addressed the point. Their sales people tried to get us to buy one of their "latest and greatest" printers. Needless to say, their "L&G" fades just the same. Interestingly, I could substitute Phaser paper for the B&W and had no problems with output, even 3 years afterwards. Sadly, color prints didn't work with the Phaser paper.
If you want high quality dye-sub prints, my suggestion is to try the Kodak line, or the Codonics (using the same print engine). The prints cost more ($2.50/sh), but then you get what you pay for...
Cheers
-------------------------- John Bonevich NIST, Metallurgy Div. B164 Gaithersburg, MD 20899 USA TEL: (301) 975-5428 FAX: (301) 975-4553
} } I also have a Sony printer (UP-D8800, 8.5"x11" dye sublimation) and } experience problems with permanence. Initial output looks good, } but after only a couple of weeks on the desk (no sunlight, just } standard fluorescent lighting) prints are noticably faded. A simple } sheet protector seems to keep that from happening, but it is } annoying to have to put everything in one. Print cost for the } UP-D8800 is about $2 per color print. I mentioned this to Sony at } the MSA meeting in Atlanta. Their only suggestion was to use a } different print paper, that accepts a laminated cover film. The cost } and description of the paper made me decide to stay with sheet } protectors, at least for now. } } Interestingly, we have a smaller format Sony (UP-1800MD, as I } recall; prints on paper ~4"x5.5") that does not have this problem. } I guess they are using different print systems (Sony--hint?) } } I haven't noticed any transfer of "ink" for either of these printers. } } Jim Passmore } Analytical Chemist } Cryovac Division, Sealed Air Corporation } P.O. Box 464 } Duncan, SC 29334 } } ---------- } } From: corwinl } } To: Microscopy } } Subject: Prints: permanence } } Date: Friday, October 09, 1998 12:04PM } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Everyone: If anyone out there in the continental US is selling a Philips CM-12 in good working order please contact: dclilly-at-LIPO.COM or gantz-at-med-biophd.bu.edu Thanks, Donald Gantz Boston Univ School of Medicine
I do not make my primary money from using microscopes, but I feel compelled to enter my 2cents worth....
If there were no vendors we would have no toys........
Being that I wish to learn as much as I can about the use of microscopes I have found many vendors extremely helpful even when we did not even have the funds to purchase something new but they were willing to take a little time to provide us with an education so we would not start off in the wrong direction.
We have been using a Phaser 450 dye-sub for about a year and haven't noticed any very obvious problems(yet) with print longevity. I encountered a Codonics for the first time earlier this week and was impressed by its paper quality, though I have yet to do a close comparison between its output and our Phaser's. Digital Imaging magazine has an article this month about the products for the fine-art digital printing market and they mention reports published by Wilhelm Imaging Research which tests the color longevity of the commonly used media. Apparently Henry Wilhelm is the expert in the science of studying the permanence of color photographs and digital media. Has anyone seen his reports and does he test dye-sub media?
I am seeking a very inexpensive but functional microtome to use in teaching histology to high school students. The school cannot afford it, so I am paying for it myself. Can anyone direct me to the source for such a device? Thanks,
I've been trying to section very small, very resistant crustacean cysts. They've been fixed in gluteraldehyde, dehydrated to 100% EtOH, infiltrated and embedded in LR White. I use a glass knife.
Semi-thin sections for LM turn out well enough, but I do need to shift the knife edge frequently. Ultra-thin sections are the problem. For a while I gave up on TEM, and thought I would make to with just LM for my study, but I really would like to get a look at the ultrastucture.
Even the first sections have rents in them where the tissue is; the sections tend to split in two when they hit the water. I had tried osmicating the tissue before I switched to room temp. polymerization with LR White. The sections would look like swiss-cheese as the tissue would drop out of the sections and sink to the bottom of the boat.
I looked at the Microscopy Tips & Tricks page re: sectioning insect eggs; I don't seem to have the same infiltration problems (thank goodness!). Much of the advice concerns softening up the eggs before embedding. I can't really try this since I'm running out of specimens and time.
I don't think I can get my hands on a diamond knife. I can't afford one and I don't know anyone willing to lend one (not surprising; I clean up my cysts really well, but there are still a few sediment grains in the mix when I embed. I've gotten good at trimming them out, though).
I think this is a hopeless case, but if there is something I can do, please please tell me!
Thanks for reading this,
Sonia McGowan Univ. of San Diego http://www.acusd.edu/~scawsey
In a recent conversation, Kodak claims that color dye sub prints made with the clear laminate coating are 30 year "permanence". Their B&W does not have the clear laminate. Ink jets are supposedly about 2 years.
My 2 cents worth.
Damian Neuberger Baxter International
At 03:05 PM 10/16/98 -0400, John Bonevich wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I've used both LR-white and lowicryl on the same samples (chick and rat aorta), and found I got better preservation with the LR-white. I went on to do IEM w/ the LR-White and got it to work. I polymerized some in a heating block in an eppendorff buried in sand, and got that to work. A friend used a PCR machine, but I don't know that you want to risk mucking up a thermal cycler for this. I'm in a different lab now with limited equipment, and was considering polymerizing it in a eppendorf in a floater in a beaker of water in an 60 deg. oven. I take it into 100% EtOH before going into LR-white, so I should have some breathing room as to having too much water in my sample to polymerize. Has anyone else done this? I know, I know, it's bad to get water into an oven used on organic resins or paraffins. This oven would be mostly devoted to this application, and I'm hoping the water would buffer temperature fluctuations. Does this make sense to anyone else? Charlie Ginsburg NCC Research Dept. Lombard IL _________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
There was a question about suppliers of second-hand microscopes on the list (march and april '98).=20
This is a summary of the answers, kindly provided by Mr. Thomas C. Isabel= l, { {tc_isabell-at-fischione.com} } .
I have visited the given websites several times: every once in a while, there is some histological equipment for sale at decent prices...
In my country (Belgium) second-hand equipment is very hard to get. At flea-markets here you can find once in a while some equipment, but it takes time: it took me some years to have decent basic equipment for my amateur-lab (microscope, microtomes, incubators etc.).
If you think that US$ 3 000 isn't to much: try to find out if there's an agent of the Dutch brand "EUROMEX" in your country. US$ 3 000 is nowadays here about the price for a manualy operated EUROMEX rotary microtome acc. to Minot (model "MIC 505"). It works well for not to large specimens (sections 10 mm * 10 mm, 4 =B5m - 25 =B5m thick; for larger sections I us= e an old "Stiassnie - Paris" vintage about 1930.). The MIC 505 is in its basic version equiped with a "real" microtome knive ("B" or "C", 17 cm). A razorblade holder or an adapter for disposable blades are optional.=20
Their sliding microtomes are in the same price range (If time is not of t= he utmost importance and one wants to cut all sorts of specimens one should consider a sliding microtome instead of a rotary...).
No financial intrest: just a statisfied user!
Some second-hand dealers in the USA: http://www.labx.com http://www.labequip.com http://www.montanamicroscope.com http://www.wwweb-pro.com/cls http://www.execpc.com/ume http://www.lehmanscientific.com http://www.capovani.com http://www.sci-equip-ex.com http://www.bid-service.com
Phone Contacts:
Martin Microscope Co (864) 242-3424 or (864) 859-2688 Bob Martin Mel Sobel 1-888-ALL-SCOPES or (516) 935-7794 Technical Instruments (415) 431-8231 Rick Staples=20 Bay Optical (415) 431-8711 Tom Henry ARC Instruments (606) 498-1345 Phil Hutcheson
A corespondent from the USA also told me "...Another place I've found tha= t is quite interesting is the auction : www.ebay.com Sometimes, they have quite reasonable microscopes and accessories listed. You are dealing with individual sellers directly ( which may have problem= s ) but there are many good 2nd hand deals.".
Applications are sought for a technician/professional level position in the x-ray scattering and microscopy facilities of the School of Materials Engineering at Purdue University. Responsibilities include operation, technical support, users training and maintenance of the x-ray powder, texture, and singlre crystal facilitiesas well as assisting in the EM and EDS components of the microstructural facility. The position requires a technician program, B.S. or M.S. degree in technology, engineering or science of x-ray and/or EM analysis. Prference will be given to applicants having experience in electrical and vacuum systems maintenance. Interested applicants should send a letter of interest and a resume along with names, addresses and telephone numbers of three references to : Said Mansour. School of Materials Engineering, 1289 Materials and Electrical Engineering Building, Purdue University, West Lafayette, IN 47907-1289 or by Fax to 765-494-1204 or by email to: said-at-ecn.purude.edu.
A colleage at our university is in need of high voltage transmission electron microscopy. Probably 300 kV would work, although 1 mV may be needed. Does anyone have info on the availability and use of such instrumentation?
Thanks, John B.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
If you use the color rolls to print b&w, then you will have the laminate. The prints cost $0.05 more, but it's worth it. It is not recommended to switch the rolls in the Kodak printer (an advantage of the Sony), so I just use color. You may have to adjust the tone curves to get the "best" b&w output.
My 5 cents worth ;-)
-------------------------- John Bonevich NIST, Metallurgy Div. B164 Gaithersburg, MD 20899 USA TEL: (301)975-5428 FAX: (301)975-5443
} Regarding print permanence: } } In a recent conversation, Kodak claims that color dye sub prints made with } the clear laminate coating are 30 year "permanence". Their B&W does not } have the clear laminate. Ink jets are supposedly about 2 years. } } My 2 cents worth. } } Damian Neuberger } Baxter International } }
Dear .edu listers, I received a back channel message from: Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901 pointing out that my "ivory tower" remark was rather inflammatory. I was replying to Ted Dunn's comments and did not realize that he was a vendor as he made no mention of the fact anywhere in his message. In his follow-up he does mention the fact. I was under the mistaken impression that he was not commercial. Bob, you're right, and I apologize for my remarks. I very much enjoy and respect my academic customers. In fact I very much enjoy and respect virtually all of my customers, most of whom I have known for about 20 years and consider to be friends perhaps more than customers. Others have more eloquently stated that there is a good deal of helpful information being presented and those who are offended by mention of commercial availability of various items and supplies should merely delete the messages without reading them and getting upset. Again, my apologies to the academic community for my hasty remarks.
Sincerely, Ken Converse owner Quality Images third party SEM service
Our lab has an ancient, but functioning International Equipment Co. Model CTD cryostat/rotary microtome. Is there anybody out there in microscopy land that might have a manual, schematic, parts list, etc. for this old unit? The cooling system works fine, but the microtome needs a couple parts. In particular, the crank seems to have a worn-out sleeve or bushing that makes turning it difficult and has generated a little pile of metal shavings over the years.
Maybe even someone has an old surplus unit that could be scavenged for parts?
And, yes, ahem, vendor replies are most welcome.
Thanks!
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Ronald LHerault wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } I am looking for a supplier of Aquembed. Aparently it is some kind of } resin that displaces water before cells are embedded in a material like } Epon 812. I can't find Aquembed in the catalogs I have. } } Thanks } } Ron } lherault-at-bu.edu
-- Dear Ron:
Ladd Research sells:
#21325 - Aquembed resin - 100 ml
#21225 - Aquembed Embedding Kit - 100 ml Aquembed resin 250 ml DDSA 30 ml DMP-30 60 ml DBP
Before anyone gets mad at us, my husband and I own Ladd Research which has been a commercial vendor of a wide variety of microscope supplies for over 40 years
Aquembed is in stock and is shipped the day it's ordered. Please contact us by e-mail or call 800-451-3406 for prices. Thanks, Rita Arnott
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
I will probably be only one of many pointing out the minor error in the posting by Damian (below), but Kodak B&W prints not only HAVE the clear laminate coating ("ExtraLife"), but it is the only way you can order those ribbons. For color prints you can either get the laminate or not (the latter recommended for making overheads). I do not in any way work for Kodak or sell their products, but am a very satisfied customer with their 8650 printers.
By the way, the color on overheads (without the extralife coating) will bleed into certain types of clear plastic sheet protectors. I believe I read one time which type of plastic reduces this problem, but no longer remember where or the details (typical!). I have been using vinyl and that is a problem after about three years of storage. Anyone have better ideas?
Cheers, John Vetrano john.vetrano-at-pnl.gov
---------- } From: "dneuberger-at-mindspring.com"-at-Sparc5.Microscopy.Com Sent: Friday, October 16, 1998 7:22 PM To: John Bonevich; microscopy-at-Sparc5.Microscopy.Com
Regarding print permanence:
In a recent conversation, Kodak claims that color dye sub prints made with the clear laminate coating are 30 year "permanence". Their B&W does not have the clear laminate. Ink jets are supposedly about 2 years.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear .edu listers, } I received a back channel message from: } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } pointing out that my "ivory tower" remark was rather inflammatory. I } was replying to Ted Dunn's comments and did not realize that he was a } vendor as he made no mention of the fact anywhere in his message. In } his follow-up he does mention the fact. I was under the mistaken } impression that he was not commercial. } Bob, you're right, and I apologize for my remarks. I very much enjoy } and respect my academic customers. In fact I very much enjoy and } respect virtually all of my customers, most of whom I have known for } about 20 years and consider to be friends perhaps more than customers. } Others have more eloquently stated that there is a good deal of helpful } information being presented and those who are offended by mention of } commercial availability of various items and supplies should merely } delete the messages without reading them and getting upset. } Again, my apologies to the academic community for my hasty remarks. } } Sincerely, } Ken Converse } owner } Quality Images } third party SEM service
We have a Codonics and have not problem switching back and forth between B&W and color rolls. Yes, we print B&W with the color roll if we want perm. but I thought I had figured it to be more expensive than 5 cents from our supplier. I'll have to check on that. The biggest reason for using the B&W rolls is speed of output.
Damian Neuberger Baxter International.
At 08:45 AM 10/17/98 -0400, John Bonevich wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I guess you missed my reply posted on Tue. 10/13/98 (around 2 am.). It's here again.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
Here is the recipe for Spurr's + Epox 812, I use. Basically, it is 1:1 mixture (by weight) of Spurr's and Epox 812. Each resin mixture must be completely homogenized, including accelerator, before combining the two.
1. Prepare Spurr's in a 100 ml disposable beaker. (Based on Spurr's original formula)
VCD -------- 10 g DER 736 ---- 6 g NSA -------- 26 g DMAE ------- 0.4 g ------------------- Total ------ 42.4 g
2. Prepare Epox 812 in a 50 ml disposable beaker. (Based on Luft's original formular, except for reduced DMP-30)
Epox 812 (WPE = 155*) --------- 25 g DDSA (MW = 266) -------------- 14 g NMA (MW = 178) -------------- 11 g DMP-30 ----------------------- 0.5 g
*If WPE is different, proportions of each component should be recalculated. Calculations can be found at http://www.emsdiasum.com/ems/techdata/68.html).
3. To make 1:1 mixture, add 42.4 g of Epox 812 mixture into the 100 ml beaker containing Spurr's and mix throughly. (Amount of Epox mixture can be reduced for hard-to-infiltrate tissues.)
4. Infiltration for plant tissues.
Transitional solvent Resin Mixture Infiltration (propylene oxide or (Spurr + 812) Time (with diethyl ehter) rotation or occasional swirring) -------------------- -------------- ------------ 3 part 1 part 2 hrs. 2 2 2 hrs. 1 3 4 hrs. 0 4 4 hrs. 0 4 6 hrs. -----------------------------------------------------------------
5. Cure blocks at 60 C for 48 hrs.
Regards,
Seung-Geuk Shin
} -----Original Message----- } From: Michelle L. Peiffer [mailto:mlk101-at-psu.edu] } Sent: Monday, October 12, 1998 12:31 PM } To: Seung-Geuk Shin } Subject: RE: Spurr resin } } } I was quite interested to see this reply, we routinely use Spurr's with no } trouble in insect tissues and cell cultures. However recently we have a } number of students without access to diamond knives who are having trouble } getting quality thin sections of plant tissue embedded in Spurr's. Would } you mind sharing your recipe for this spurr's epox mixture? Thanks . } } } #################################################### } Michelle Peiffer } Electron Microscope Facility for the Life Sciences } The Biotechnology Institute for Research and Education } 1 South Frear Lab } University Park, PA 16802 } 814-865-0212 email:mlk101-at-psu.edu } #################################################### } } }
Does anyone have experiences on the better procedure of staining of ABS(Poly acrylonitril-butadiene-styrene) for TEM? I appreciate any kind of help. Ren-Jye
One of the several types of resin we use in our laboratory is Spurrs resin. ( I know it is highly toxic and we use extreme care when handling it). However, my question has to do with the oven we polymerise the resin in. In other laboratories I have worked in we always used a small copper oven with no special whiz bang characteristics, just a knob for setting the required temperature. These copper ovens had an outlet hose which extracted the fumes into a fume hood preventing the subsequent polymerisation of these fumes on the inside of the oven. I have been unable to find the supplier of these ovens as the ovens are all extremely old. Our current oven is useless as the resin has successfully gummed up the interior. We have never spilt resin in the oven but do polymerise all waste containers including gloves and anything else the resin has been in contact with. What sort of ovens are everyone else using and has anyone got a suggestion on where we may find a simple polymerising oven. We do not want to spend large amounts of money on an oven with 'reinforced triple glass windows', or 'Microcomputer PID control', or with any other totally irrelevant feature not necessary for the polymerisation of Spurrs. We have a sensitive oven for use with LR White resin which we will not put Spurrs in. Our LR White resin specimens are sealed from air and sit in an aluminium block which is wrapped in foil. These specimens are therefore very clean and do not contaminate our 'good' oven. Waste LR White is polymerised in the 'Spurr' oven.
thanks in advance,
Sarah Ellis
Trescowthick Research Centre Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne 8006 Victoria Australia
I got a copy of Image Tool by internet and there I saw a message to subscribe in IT-list. Now I received a message saying that it do not exist. Someone knows how can I discuss with users of ImageTool? Thanks, Rejane Galindo
Rejane Pimentel Galindo Universidade Federal Rural de Pernambuco Av. Boa Viagem, 6592/602 51130-000, Recife, Pernambuco, Brasil ggalindo-at-elogica.com.br =46ax (081) 441 4697
You asked for it. The following news story I received last week is too funny not to post to the list, despite its lack of microscopy content (for which I apologize in advance).
According to the Knight-Ridder News Service, the inscription on the metal bands used by the U.S. Department of the Interior to tag migratory birds has been changed. The bands used to bear the address of the Washington Biological Survey, abbreviated "Wash. Biol. Surv.", until the agency received the following letter from an Arkansas camper:
"Dear Sirs: While camping last week I shot one of your birds. I think it was a crow. I followed the cooking instructions on the leg tag and I want to tell you, it was horrible."
The bands are now marked Fish and Wildlife Service.
I am searching for any recent (within 5 - 10 years) advances in the use of Scanning Electron Microscopy in the field of Fractography. This information is for a term paper for an SEM graduate school course. Any useful information or suggestions for additional locations for information would be appreciated. Thanks, Frank Watson eCarolina Products Plant email: frank.watson-at-lighting.ge.com voice: 8*565-5177 (outside GE (919) 731-5177) Fax: 8*565-5114 (outside GE (919) 731-5114)
May I suggest you either read or, better still, purchase my book "Low Temperature Microscopy and Analysis" published by Plenum Press New York in 1992
Patrick Echlin Cambridge University
On Fri, 16 Oct 1998 reznor-at-holly.colostate.edu-at-Sparc5.Microscopy.Com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Email: reznor-at-holly.colostate.edu } Name: Glen El-Hayek } School: Colorado State University } } I was wondering if there was anywhere I could find info on what exactly } freeze fracture electron microscopy is and how it works. I am working on a } web site for a cell bio class and that is the topic we have to research. } } Thank you very much, } Glen El-Hayek } Colorado State University } } } }
} Date: Tue, 13 Oct 1998 13:25:24 -0400 } From: Ronald Anderson {anderron-at-us.ibm.com} } To: microscopy-at-sparc5.microscopy.com } Subject: Fonts in Messages, etc. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I agree that all messages should be in plain text. } } I especially do not like messages sent as attachments that have to be opened. } This seems to be happening more frequently of late. } I automatically delete all such msgs, unopened. } } Ron } } Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com } IBM Analytical Services; http://www.chips.ibm.com/services/asg } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
John, We have both a 1.2 mev HVEM and a 400kv IVEM at our Biological Microscopy and Image Reconstruction Resource (BMIRR) in Alnany NY. They are both available for use as a NIH resource. The HVEM is an AEI EM7 and is routinely used for thick section (0.5 to 2 micron or more depending on the mass thickness) microscopy including tomography and stereo reconstruction techniques. The IVEM is a JEOL JEM4000FX is routinely used for automated tomography and automated low dose microscopy. Both have televiewing capability using a web browser.
Contact: Dr. Conly L. Rieder (Phone: (518) 474-6774, Email:rieder-at-wadsworth.org ) to apply to use our resource or connect to our web site at:
www.wadsworth.org/spider_doc/bmirr/index.html
Dave
******************************************* David Barnard HVEM operator Wadsworth Center New York State Dept. Health Albany, NY 12201-0509 barnard-at-wadsworth.org (518) 473-5299 *******************************************
In previous email correspondence it was suggested by few people to use Evans Blue stain as a counterstain on FITC labelled sections. Is there a special protocol for it. Also, will it work with CY2 labelled sections? And would it fluoresce with CY3 filter? Thank you for your help, Lilith
------------------------- Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We also have a Codonics printer and it is my understanding that the B & W has a transparent laminate added on. This is done when it pulls the paper back into the printer after the initial print is produced and before releasing it into the pick up tray. You can actually see the positioning of the laminate on the paper. You have a choice of using color ribbons with or without the laminate sheet which does affect the cost of the color print. I believe the sheet is the same chemistry for both B&W and color and protects against UV color change as well as effects of air components. Can anyone confirm this.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------
John,
We have a Codonics and have not problem switching back and forth between B&W and color rolls. Yes, we print B&W with the color roll if we want perm. but I thought I had figured it to be more expensive than 5 cents from our supplier. I'll have to check on that. The biggest reason for using the B&W rolls is speed of output.
Damian Neuberger Baxter International.
At 08:45 AM 10/17/98 -0400, John Bonevich wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
RFC822 header -----------------------------------
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I would like to find out what type of mag. calibration schedule you might be using, if at all. Is it every 3mnths, 6mnths, or every month? Dictated by ISO? I do not wish to start another thread about parameters effecting mag. calibration but more on how often this calibration is performed.
TIA
David Rose W.L. Gore and Associates Elkton, MD 21921 410-506-2958
Hi everyone. I'm fixing cell suspension cultures for TEM for an undergraduate research project. I've tried embedding the cells in agar and using agar chunks to process for EM... didn't like that too much. Now I'm just fixing cell pellets of the appropriate size. With enough g's the cells will stick to form a nice, detachable pellet (especially after fixing w/ glut.)
Here's my question: Is it the more common practice to form a nice, tight pellet of cells right away and fix that, OR is it best to resuspend the cells into a suspension after EVERY step (glut, rinsing, OsO4, dehydration, etc...) and only forming a compact pellet in the very final infiltration stages?
In my experience so far, pellets formed at the end stages (rather than during primary fixation) don't tend to stay together... and centrifuging through resin has it's difficulties.
Most of the papers I'm looking at are VERY lax in their description of EM prep methods. So, to the experts out there, what is generally assumed when a paper says something like, "Cell suspensions were fixed as usual in 2% glut. and 1% OsO4 and embedded in resin."
This was an interesting topic for us as we are A2LA accredited. We initially wrote into our quality program that we verify the magnification of our optical microscopes annually. Our SEM is calibrated each session requiring critical measurements.
A2LA's position was that the magnification of the optical microscopes does not change from the factory settings, hence they never need calibration. We continued to have our optical service man calibrate and clean the microscope on an annual basis until last year. A2LA now requires all outside calibration organizations to be A2LA accredited themselves. This eliminated our regular service engineer. Our new A2LA accredited calibration organization is not accredited to verify magnification, hence our annual magnification has to be deleted from our quality program.
Alan Stone ASTON Metallurgical Services
At 01:47 PM 10/19/98 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Laszlo Komuves wrote: } } In your response to a question about Aquembed resin, you mentioned that } you carriy this resin. } However, I am not familiar with this resin. Could you send me some } information/references why would anyone use this resin, wat are the } advantages, etc.
Aquembed is very low vescosity and water soluble so you can dehydrate and embed in the same material. It is highly purified.
} Is this a repackaged version of Spurr's?
No, Aquemebed is not a repackaged version of Spurr, it is completely differnt.
I recall a paper publisehed few } years ago in Microscopy Research and Technique suggesting not to complete } dehydration and polymerize at 40oC to preserve antigenicity in Spurr's } resin.
This would also be problem with Aquembed since it is meant to be polymerized at a higher temperture.
} Sincerely, } Laszlo G. Komuves, Ph.D.
If you have further questions you can contact me or call our chemist, Dr. Charles Duvic, directly at 1-800-451-3406.
Thank you for your interest,
Rita Arnott
Disclaimer: As stated previously, Ladd Research sells Aquembed. --
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
In case this discussion has not come up yet on the server, here it goes. We are running a Jeol JSM 6400 with Link eXL spectrometer and have no info from supplier or manufacturer about year 2000 compliance. Any ideas/input out there. I'm sure many of us are in the same boat (maybe without a paddle).
We go through a major calibration every 6 months although I am not sure where that timeframe came from. I believe it is up to the operator to decide when the instrument requires calibration and that mey be deduced by monitoring results.
I would also be interested in hearing what everyone considers as passing the calibration. Is it +- 5%, 10% and what is it based on?
} Now I received a message saying that it do not exist. Someone knows how can } I discuss with users of ImageTool?
Sorry, but I can't resist...I will write in Portuguese (It's the first time I find a msg from someone who, probably, speak Portuguese)
Finalmente alguem que fala (penso eu!) Portuges!!!
Tambem gostaria de saber se ha alguem a trabalhar com o Image Tool!!
Parece ser um programa ao nivel de alguns comerciais no entanto nunca vi nenhuma discucao acerca disso. Tambem ja enviei algumas mensagens mas nunca tive FeedBack!!
Se receber alguma resposta diga-me alguma coisa (se for possivel)
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all,
As a total amateur at SEM (almost) I was wondering if there was someone = out there who could give me a private run through of what is now available.= The last time I looked at SEM's was over 10 years ago and things seem to = have moved along almost as fast as in the computer industry (although I = haven't seen laptop SEM's yet).
This subject is probably not appropriate for the list, so it would be good = if volunteers contacted me and didn't post for general consumption.
My field is biological research so I would be particularly interested in = what imaging possiblities there are for cells, tissues and isolated = fractions (or even proteins).
Many thanks in advance,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org
(InterLock SMTP Gateway 3.0); Mon, 19 Oct 1998 15:44:00 -0700 Received: by tmpil001.tmp.allied.com (Internal Mail Agent-1); Mon, 19 Oct 1998 15:44:00 -0700 Message-Id: {c=US%a=_%p=ALLIED%l=ALLIED/NAAERO1/0097A961-at-tmpcn541.wins.allied.com} "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-Sparc5.Microscopy.Com}
David,
We verify the mag calibration on our SEM's once a year. That is unless the high voltage supply or condenser lenses, etc. were malfunctioning and had to be repaired or replaced. ASTM E766 recommends the frequency of the mag calibration to be determined by the user.
Harry Ekstrom
---------- } From: "drose-at-wlgore.com"-at-sparc5.microscopy.com To: Microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
Dear Rejane, Unfortunately, the IT-list seems to have died and the Image Tool software site has not been updated for over a year, now. I think maybe the wonderful fellows who wrote it were laid off. Good luck. You wrote: } } I got a copy of Image Tool by internet and there I saw a message to } subscribe in IT-list. } Now I received a message saying that it do not exist. Someone knows how can } I discuss with users of ImageTool? } Thanks, } Rejane Galindo } } Rejane Pimentel Galindo } Universidade Federal Rural de Pernambuco } Av. Boa Viagem, 6592/602 } 51130-000, Recife, Pernambuco, Brasil } ggalindo-at-elogica.com.br } Fax (081) 441 4697 } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Many thanks to several people who helped me with my search for Long Working Distance Ojectives. I have found a source for them. -- ********************************************************************** Philip Datner datner-at-netcom.com San Jose, CA datner-at-engmail.ulinear.com **********************************************************************
We have exactly the same setup, JSM6400 + Link eXL. I have contacted both JEOL and OXFORD about Y2000 bug.
SEM has no date support, so it fully Y2000 compliant.
OXFORD distributes the Year 2000 Statement describing in details what is going to happen to the system in the Y2000. I believe they'll send you one if you request it. In brief, the overall system will be "fit for purpose" after the rollover.
Alexander Titkov
Millennium Inorganic Chemicals PO Box 245 Bunbury WA 6231 Australia Ph: (08) 9780 8505 FAX: (08) 9780 8500 E-mail: atitkov-at-micl.com.au
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In case this discussion has not come up yet on the server, here it goes. We are running a Jeol JSM 6400 with Link eXL spectrometer and have no info from supplier or manufacturer about year 2000 compliance. Any ideas/input out there. I'm sure many of us are in the same boat (maybe without a paddle).
id AA10188; Tue, 20 Oct 1998 10:02:26 +0800 Received: by ucl.itri.org.tw(Lotus SMTP MTA v4.6.1 (569.2 2-6-1998)) id 482566A3.000BEBC9 ; Tue, 20 Oct 1998 10:10:12 +0800 X-Lotus-Fromdomain: UCL To: Microscopy-at-Sparc5.Microscopy.Com Message-Id: {482566A3.000AD2E7.00-at-ucl.itri.org.tw}
I am looking for the TEM sample preparation of collagen, including staining...... Can anyone share experiences or provide references on this subject? Thanks in advance. Ren-Jye
In the past I have more or less let the cells tell me how they like to be treated. Some cell types will form a nice, tight pellet after a single spin in the primary fixative. If this is the case, I usually leave them alone from this point on if possible, and treat the pellet like a small piece of tissue. I only spin the cells if it is absolutely necessary.
There are other cell types which, no matter what you do to them, refuse to stay in a pellet. These are the ones which I usually embed in either agar or low-melt agarose.
You can pellet your cells at very high speed using an airfuge centrifuge in proper fixative and continue post fix and rest of the dehydration and embedding with the pellet. The pellet should not fall apart. This works quite well. You can embed the pellet by breaking it into small pieces after the final stage of infiltration.
Good luck
Soumitra Ghoshroy Ph.D. Department of Plant Sciences University of Arizona 303 Forbes Building Tucson, AZ 85721 Tel: 520-621-1230 Fax: 520-621-7186 e-mail: ghoshroy-at-ag.arizona.edu
On Mon, 19 Oct 1998, Douglas Matthews wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi everyone. I'm fixing cell suspension cultures for TEM for an } undergraduate research project. I've tried embedding the cells in agar and } using agar chunks to process for EM... didn't like that too much. Now I'm } just fixing cell pellets of the appropriate size. With enough g's the cells } will stick to form a nice, detachable pellet (especially after fixing w/ glut.) } } Here's my question: Is it the more common practice to form a nice, tight } pellet of cells right away and fix that, OR is it best to resuspend the } cells into a suspension after EVERY step (glut, rinsing, OsO4, dehydration, } etc...) and only forming a compact pellet in the very final infiltration stages? } } In my experience so far, pellets formed at the end stages (rather than } during primary fixation) don't tend to stay together... and centrifuging } through resin has it's difficulties. } } Most of the papers I'm looking at are VERY lax in their description of EM } prep methods. So, to the experts out there, what is generally assumed when } a paper says something like, "Cell suspensions were fixed as usual in 2% } glut. and 1% OsO4 and embedded in resin." } }
Quite a lot depends on the user environment, paper etc... I did quite a exstensive test. When we were shopping for printers I contacted all the suppliers. It took quite a lot of convincing for to get themto bring their demo models over for a test drive. We tested for print permanence as well. The prints were left in direct sunlight with half of the print shaded off. This was done over a period of four weeks. All inkjet prints faded irrespective of printing media. All dyesublimation prints survived without any noticeable fading. (We only had excess to Kodak and the Spectra starr duyesublimation printers) A Africa logistic problem at the time and continuing!
} experience problems with permanence. Initial output looks good, } but after only a couple of weeks on the desk (no sunlight, just } standard fluorescent lighting) prints are noticably faded. A simple } sheet protector seems to keep that from happening, but it is } annoying to have to put everything in one. Print cost for the } UP-D8800 is about $2 per color print. I mentioned this to Sony at } the MSA meeting in Atlanta. Their only suggestion was to use a } different print paper, that accepts a laminated cover film. The cost } and description of the paper made me decide to stay with sheet } protectors, at least for now. } } Interestingly, we have a smaller format Sony (UP-1800MD, as I } recall; prints on paper ~4"x5.5") that does not have this problem. } I guess they are using different print systems (Sony--hint?) } } I haven't noticed any transfer of "ink" for either of these printers. } Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
It depends entirely on how anal you want to get (no offense intended). Yearly calibration checks, properly documented, will satisfy any certification demands that I am aware of. If a certifying body is concerned, i.e. ISO 9000, I usually suggest that a customer own and document the storage and use of their own standard - current best in US is NIST SRM-484. I carry one, but due to the environmental changes and mechanical abuse it goes through, I prefer not to rely on it where questions may arise, although NIST certifies the calibration unless it requires excessive polishing.
When any repairs or alterations are made to any portion that may affect the calibration, a new certification should be performed. This would include any changes to the accelerating voltage, condensor lens, objective lens, record CRT, scan generator and mag readout sections, as well as any reference supply or power supplies that feed those areas. Digital image capture devices usually include a simple means of adjusting their calibration should any changes be made to them.
Owning your own standard also allows you to establish your own SPC controls on the instrument by tracking the shorter term flunctuations in calibration. Just be sure that you follow the same procedures used in the yearly calibrations, i.e. degaussing the column and using the same calibration point.
In most cases, manufacturers will specify calibration within 5 - 10%. This is because that calibration must track over the entire operating range of the instrument - primarily working distance, condensor and accelerating voltage ranges.
I advise that a single point be chosen, close to the normal operating range of the instrument. For example, a particular instrument may normally be operated at 12mm working distance, 20KV and 2A condensor current for concurrent EDS analysis. We will choose that point as the calibration point and always make calibration at that point. While the instrument will be maintained to be be within manufacturer's specs throughout its operating range, at that single point we can keep it within 1 or 2%.
The customer will be aware that the calibration certainty will decrease as they move away from that point. However, the use of a single, high certainty point, allows them to easily track instrument performance with some accuracy. Obviously, using only manufacturer's specifications, it would be hard to perform checks over hours or days that would have any real meaning. Using a single point calibration of high certainty allows one to accurately measure the variation over hours or days and thus determine with good statistical accuracy the performance of the instrument. The variation you measure can give you some idea of the accuracy and repeatability of your instrument. A large or changing variation likely points to repairs that need to be made.
} Hello List, } } I would like to find out what type of mag. calibration schedule you } might be using, if at all. Is it every 3mnths, 6mnths, or every } month? Dictated by ISO? I do not wish to start another thread } about parameters effecting mag. calibration but more on how often } this calibration is performed. } } TIA } } David Rose } W.L. Gore and Associates } Elkton, MD 21921 } 410-506-2958 } } } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Despite manufacturer's claims, there is little that is new in SEMs. After nearly 20 years in development, the environmental SEM is practical, and probably has much to offer in your discipline. Basic resolution has generally been cut in half over that time period (ETEC spec'd their SEMs at 75A 20 years ago, and did so with a measure that seems more conservative than today's specs) but FE instruments have gone farther, although I don't know of any real use for FE in biological applications (y'all let me know if I'm wrong).
Your question is very germain to this group, I would think. I would love to hear if anyone has any additional improvements they have noticed that I haven't mentioned. Of course I will conceed to improvements in vacuum system components, reduced electronic part counts and, of course, the introduction of digital image capture and processing. All important, however, not producing any real improvement on the basic imaging capabilities.
} Dear all, } } As a total amateur at SEM (almost) I was wondering if there was } someone out there who could give me a private run through of what is } now available. The last time I looked at SEM's was over 10 years } ago and things seem to have moved along almost as fast as in the } computer industry (although I haven't seen laptop SEM's yet). } } This subject is probably not appropriate for the list, so it would } be good if volunteers contacted me and didn't post for general } consumption. } } My field is biological research so I would be particularly } interested in what imaging possiblities there are for cells, tissues } and isolated fractions (or even proteins). } } Many thanks in advance, } } Paul Webster, Ph.D } House Ear Institute } 2100 West Third Street } Los Angeles, CA 90057 } phone:213 273 8026 } fax: 213 413 6739 } e-mail: pwebster-at-hei.org } } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
The Lehigh Short Course to be run in early June 1999 will teach you everything you ever wanted to know about SEM and XRMA.
Contact Sharon Coe at e-mail address {slc6-at-lehigh.edu} for more information.
Patrick Echlin Cambridge University (Teacher on the Lehigh Course)
On 20 Oct 1998, Paul Webster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } } As a total amateur at SEM (almost) I was wondering if there was someone out there who could give me a private run through of what is now available. The last time I looked at SEM's was over 10 years ago and things seem to have moved along almost as fast as in the computer industry (although I haven't seen laptop SEM's yet). } } This subject is probably not appropriate for the list, so it would be good if volunteers contacted me and didn't post for general consumption. } } My field is biological research so I would be particularly interested in what imaging possiblities there are for cells, tissues and isolated fractions (or even proteins). } } Many thanks in advance, } } Paul Webster, Ph.D } House Ear Institute } 2100 West Third Street } Los Angeles, CA 90057 } phone:213 273 8026 } fax: 213 413 6739 } e-mail: pwebster-at-hei.org } } }
I have a problem in resolution of Scanning Electron Microscope S120, Cambridge Instrument. I have tried cleaning the SEM column, as I do everytime when there is a astigmatic image. But, recently I am unable to get good image at even as low as 1000X. What might but the problem?
Thanking you all in advance.
Yours faithfully,
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road, Pune - 411004, India
this is very urgent!!! I need information/prices/quotation for gold sputter/carbon coater device for SEM samples preparation.
I would like to hear suggestions or experience of users about what to buy, suppliers, models, etc.
I appreciate very much your help and assistance.
Many thanks in advance.
Silvia Montoro Centro Regional de Investigacion y Desarrollo Guemes 3450 3000 Santa Fe Argentina e-mail: csedax-at-arcride.edu.ar fax: +54 - 42 - 550944
Our lab is ISO certified. ISO does not set predetermined cal schedules, but monitors your records and procedures to confirm that you are doing what you committed to do and are keeping the appropriate records.
The number of variables affecting SEM mag calibration are infinite. Since I need to do more than constantly check mag, my procedures commit to a single (most typical settings) mag check on a monthly basis. Complete records are maintained. This ensures that (it is unlikely) large changes in mag have not occured. Also in my procedure is the option that for any job that requires tight control of magnification, a mag check is made at the time of the job. This mag check must be made using set-up parameters similar to those which will be used for the job (W.D, kV., etc).
The introduction of low voltage SEM operation in in our lab has greatly increased our ability to image radiation-sensitive low-Z biological material, due to the reduction in penetration and radiation damage by the beam. Under these conditions FE instruments offer real benefits in resolution, even for biologists. In our facility we have only a LEO 982 FESEM, so I can't directly compare our recent results with what we would see using a thermionic emitter (W or LaB6), but I know that at low voltage in this FESEM we are able to cell structures (e.g., microvilli, microtriches, cytoskeletal and contractile filaments) with almost no coating, and far less charging than we used to see in our old FESEM, which only operated well at higher voltages.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
We jet spray volcanic ash particles (~10 microns or less) onto carbon double stick tabs for SEM/EDS study. It is easy to produce a nice distribution of particles with few that overlay one another. Problem is that later, during EDS and backscatter imaging, when the beam current is higher, the carbon tape is often damaged. It curls and bubbles thereby distorting the images.
We tried to use carbon paint but it is tricky to get the right amount spread onto the mount or it dries too quickly. Results were unfavorable.
I'd really like to have a durable, very smooth, low Z surface, that is insensitive to beam current and is easy to apply and work with in the SEM. Oh, it should also be economical too.
Any ideas? TIA
Owen
============================= Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
The Athena mineral database allows searching by chem elements, but not by actual analysis. It may be of some use.
the URL is:
http://un2sg4.unige.ch/athena/mineral/search.html
Good luck Mike
================================================ Michael L. Boucher Sr. mboucher-at-isd.net WEBPAGE http://www.isd.net/mboucher ================================================
I have contracted with XK, Inc, http://www.xk.com/company.html, to develop an application similar to what you describe. Associations between an "unknown" and members of the database will be made via text based relational searches, "best fitting" of spectra, or quantitative results depending on the materials category. Regarding your particular needs, you should contact them directly. Dennis.
________________________________________________________ Dennis C. Ward voice: 202-324-2982 FBI fax: 202-324-4018 Microanalysis Laboratory e-mail: DCWard-at-concentric.net
Hi. I know of a superb project that has developed what you seek. I doubt they will be willing to hand out the database as it is an integral part of a system they have taken many years to develop, the QemScan. Perhaps you should consider gaining access to a QemScan. Here is their contact info; paul.gottlieb-at-minerals.csiro.au or nobody-at-cat.csiro.au
Craig Harris Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
Manuel: en nuestro laboratorio utilizamos image tool frecuentemente, si tu quieres, podemos comenzar a analizar sus pro y sus contras en conjunto.. espero tu respuesta...
On 19 Oct 98 at 22:36, Manuel Norberto Valente de So wrote:
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- } ---. } } } } } } Now I received a message saying that it do not exist. Someone knows ho= w can } } I discuss with users of ImageTool? } } Sorry, but I can't resist...I will write in Portuguese (It's } the first time I find a msg from someone who, probably, speak } Portuguese) } } } Finalmente alguem que fala (penso eu!) Portuges!!! } } Tambem gostaria de saber se ha alguem a trabalhar com o Image Tool!! } } Parece ser um programa ao nivel de alguns comerciais no entanto } nunca vi nenhuma discucao acerca disso. Tambem ja enviei algumas } mensagens mas nunca tive FeedBack!! } } } Se receber alguma resposta diga-me alguma coisa (se for possivel) } } } Norberto } } (mnvs-at-ciaac.aac.uc.pt) } } } =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D Fernando D. Balducci Laboratorio de Microscopia Electr=F3nica Facultad de Ingenier=EDa - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
Owen asks .... } } } Good morning: } } We jet spray volcanic ash particles (~10 microns or less) onto carbon } double stick tabs for SEM/EDS study. It is easy to produce a nice } distribution of particles with few that overlay one another. } Problem is that later, during EDS and backscatter imaging, } when the beam current is higher, the carbon tape is often damaged.
The carbon tape is successful with distributing the effects of charging but not heat. I, at least, can only suggest you sputter with a heat conducting metal ... e.g., Al, Au, Cu ... most any metal would help, but I realize it would superimpose spectra and degrade your BSE contrast. Short of that, you'd need to lower the energy in your beam with either lower beam currents or accel voltage. I'll also be looking for other ideas ... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Please note that JEOL does have a Y2K statement on our Homepage at http://www.jeol.com along with two contacts for more information, one of whom is a Director and the other who is devoting full time to this problem. This person (corey-at-jeol.com) can give you the current status of any JEOL instrument.
Regards Steve Hamilton Marketing Manager JEOL USA, Inc. hamilton-at-jeol.com
} -----Original Message----- } From: Wayne England [mailto:wengland-at-ortech.on.ca] } Sent: Monday, October 19, 1998 4:44 PM } To: micro. listserver } Subject: year 2000 and SEMs } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } In case this discussion has not come up yet on the server, here it goes. } We are running a Jeol JSM 6400 with Link eXL spectrometer and have no info } from supplier or manufacturer about year 2000 compliance. Any ideas/input } out there. I'm sure many of us are in the same boat (maybe without a } paddle). } } Wayne England } wengland-at-ortech.on.ca } }
If you want 1.) to have a resource for researchers to look at past projects in your lab to help them form experimental ideas and know who to ask for help and 2.) to and have a clear record of importance of your facility to the institution and your essential role in it to assure future support, you really need this bibliography. Also, we attempted to keep track of successfully funded grant applications that proposed projects with our facility, but we gave up because this was too difficult.
-------------------------------------------- Michael Cammer email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 http://www.ca.aecom.yu.edu/aif/ --------------------------------------------
On Fri, 9 Oct 1998, Rick L Vaughn wrote: } labs). How many labs have to keep that information? I don't believe a } core facility should be expected to see that an experiment gets } published. I feel lucky if they remember to acknowledge the lab. Plus, } work done as abstracts, thesis, presentations, and posters are also } important. Am I alone here?
The earth is a sphere. A quarter is a disk. If you're going to lay the quarters on the surface of the sphere, the answer is very different. ANyhow, do you think this merritted a serious answer or do you think somebody is trolling with nasty bait?
-------------------------------------------- Michael Cammer email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 http://www.ca.aecom.yu.edu/aif/ --------------------------------------------
On Wed, 14 Oct 1998, Alwyn Eades wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I apologize for the tone but I feel that this was a terrible answer to send } out. I assume that the person who asked the question is either young or } not too familiar with the ideas of science. Therefore to imply that the } magnification requested can be known so accurately (more accurately than we } know any of the fundamental constants, for example) is leading the reader } astray, to put it mildly. If the diameter of a quarter varies by, say, a } part in a thousand from one to another, the answer would be 574 times ten } to the power six i. e. 574 million. All the other figures are wrong. } } Alwyn Eades } } At 09:04 AM 10/14/98 -0400, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Earth Diameter = 12,756.28 KM -at- Equater } } Quarter Diameter (US) = 7/8 inch } } } } The Earth is 573,960,854.9 times larger, so that's your mag. } } } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvannia 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu } }
Thanks to all who responded to my query on Spurr's resin. -- Regards, Gregory Rudomen Technical Specialist University Microscopy Imaging Center State University of New York at Stony Brook 516-444-3126 Greg-at-umic.sunysb.edu ********************************************************** Standard disclaimer: The opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. **********************************************************
Dear Owen, } } We jet spray volcanic ash particles (~10 microns or less) onto carbon } double stick tabs for SEM/EDS study. It is easy to produce a nice } distribution of particles with few that overlay one another. Problem is } that later, during EDS and backscatter imaging, when the beam current is } higher, the carbon tape is often damaged. It curls and bubbles thereby } distorting the images. } } We tried to use carbon paint but it is tricky to get the right amount } spread onto the mount or it dries too quickly. Results were unfavorable. } } I'd really like to have a durable, very smooth, low Z surface, that is } insensitive to beam current and is easy to apply and work with in the SEM. } Oh, it should also be economical too. } I would like to suggest Berylium as a possible solution for a smooth, low-Z surface. It can be evaporated onto the specimen and it satisfies all the given criteria. HOWEVER, there are very serious safety issues involved. If you have access to the appropriate equipment--a dedicated vacuum evapora- tor with exhaust vented appropriately, the equipment to produce small Be pellets or wire for evaporation, a safe place to clean out the bell jar, etc.--you might want to consider using Be. I think that solid Be metal is not too dangerous, and that finely-divided metal or compounds are the most hazardous. I have handled BeO powder (using a hood, of course) and had no trouble, and I heard a presentation at MSA some years ago by Dr. Hall (whose first name escapes me) where Be coating was used instead of C for low temper- ature work (where Be's conductivity is much greater than C's). With the stated safety reservations, perhaps you could produce a Be equivalent of double-stick tabs. Good luck. Yours, Bill Tivol
Ihave quite often used conductive carbon tape for such exams with no problem.
..Wonder what beam current and voltage you are running? I presume you are carbon coating the ash/tape to aviod charging and that is not the distortion
problem.
Woody White McDermott Technology, Inc. ______________________________ Reply Separator _________________________________
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Good morning:
We jet spray volcanic ash particles (~10 microns or less) onto carbon double stick tabs for SEM/EDS study. It is easy to produce a nice distribution of particles with few that overlay one another. Problem is that later, during EDS and backscatter imaging, when the beam current is higher, the carbon tape is often damaged. It curls and bubbles thereby distorting the images.
We tried to use carbon paint but it is tricky to get the right amount spread onto the mount or it dries too quickly. Results were unfavorable.
I'd really like to have a durable, very smooth, low Z surface, that is insensitive to beam current and is easy to apply and work with in the SEM. Oh, it should also be economical too.
Any ideas? TIA
Owen
============================= Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
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POSITION AVAILABLE: Director, Microscopy and Imaging Center Texas A&M University
Texas A&M University is seeking applications and nominations for Director of the Microscopy and Imaging Center (MIC). The MIC is a faculty-driven facility supporting diverse interdisciplinary teaching and research. It has a sizeable annual budget for technical and service support for multiple electron microscopes and advanced light microscopy instrumentation. Applicants must be nationally and internationally recognized scholars with strong leadership and interpersonal skills and a vision for the future of microscopy and imaging technologies. An understanding of the multi-faceted approaches to contemporary and emerging imaging technologies as well as an active research program engaged in the development and/or novel application of imaging technology will be necessary. Additionally, evidence for an appreciation of the opportunities to further enhance an interdisciplinary research environment that encourages scientific exchange between materials scientists, engineers, and life scientists related to these technologies is required. The Director will have an earned Ph.D. with expertise in the life sciences area and will have a faculty appointment at the tenured Professor rank in one or more life science departments and credentials commensurate with an Endowed Chair position.
Applicants should send a complete C.V., statement of research interests and names of five references no later than January 1, 1999. All correspondence should be addressed to:
Microscopy and Imaging Center Director Search Attn: Terry Thomas, Search Committee Chair Department of Biology, Texas A&M University College Station, Texas 77843-3258
Texas A&M University is an equal opportunity employer.
POSITION AVAILABLE: Director, Microscopy and Imaging Center Texas A&M University
Texas A&M University is seeking applications and nominations for Director of the Microscopy and Imaging Center (MIC). The MIC is a faculty-driven facility supporting diverse interdisciplinary teaching and research. It has a sizeable annual budget for technical and service support for multiple electron microscopes and advanced light microscopy instrumentation. Applicants must be nationally and internationally recognized scholars with strong leadership and interpersonal skills and a vision for the future of microscopy and imaging technologies. An understanding of the multi-faceted approaches to contemporary and emerging imaging technologies as well as an active research program engaged in the development and/or novel application of imaging technology will be necessary. Additionally, evidence for an appreciation of the opportunities to further enhance an interdisciplinary research environment that encourages scientific exchange between materials scientists, engineers, and life scientists related to these technologies is required. The Director will have an earned Ph.D. with expertise in the life sciences area and will have a faculty appointment at the tenured Professor rank in one or more life science departments and credentials commensurate with an Endowed Chair position.
Applicants should send a complete C.V., statement of research interests and names of five references no later than January 1, 1999. All correspondence should be addressed to:
Microscopy and Imaging Center Director Search Attn: Terry Thomas, Search Committee Chair Department of Biology, Texas A&M University College Station, Texas 77843-3258
Texas A&M University is an equal opportunity employer.
To all that has interest, To respond to all of our customers demands for a new catalog we are pleased to announce the release of catalog XIII. Catalog XIII promises to meet the demands of all of the microscopy market as well as histology and general biological research. We have expanded the catalog to include a unique arrangement with Diatome, the Premium Diamond Knife manufacturers as well as a complete new chapter on Digital Imaging. For a copy of this resource guide please call, write, or E-Mail us direct at Electron Microscopy Sciences, 321 Morris Road P.O Box 251 Fort Washington Pa 19034. Tel:215-646-1566 E-Mail: SGKCCK-at-aol.com. We look forward to hearing from you. Sincerely, ELectron Microscoph Sciences
I'm having a strange problem in saving high definition images onto the hard disk. This has only recently occurred. Yesterday, the first high definition image that I attempted to save took around 8 minutes, after which the system froze on me preventing me to save any further high definition images. The situation now is that I am only able to save standard definition images, but not high definition images. When the system freezes there is a warning sign that says,
"Server Cause a General Protection Fault in Module SERVER.EXE at 0001:OD89"
Rebooting the computer does not solve anything. Defragmenting the hard drive did not help.
I use SEM Philips XL20 and software V 5.21. The computer is using Windows 3.1 operating system and has an abundance of hard disk space.
The Santa Clara Valley Chapters of ASM and IEEE Reliability Society, with generous support from FEI Company, Micrion Corp, Schlumberger, and Nissei Sangyo America present:
NANOSCALE CHARACTERIZATION WITH THE FOCUSED ION BEAM Speaker: Prof. Robert Hull Department of Materials science and Engineering University of Virginia
ABSTRACT: Nanoscale characterization techniques using the focused ion beam (FIB) instrument will be reviewed. Dr. Hull will describe FIB-based techniques (either stand-alone, or in conjunction with transmission electron microscope imaging) for stress mapping in crystalline structures, dopant mapping in semiconductors, three dimensional image reconstruction, and ultra-high resolution secondary ion mass spectroscopy maps and volume reconstructions. He will also summarize additional techniques described in the literature, including FIB-induced optical emission spectroscopy and voltage contrast imaging. Finally specimen modification and damage artifacts created by the FIB beam will be discussed.
BIOGRAPHICAL SKETCH Robert Hull is an Associate Professor, and Doris and Heinz Wilsdorf Distinguished Research Chair, in the Department of Materials Science and Engineering at the University of Virginia. Prior to joining UVA he was a member of Technical Staff in the Physics Research Division of Bell Laboratories for seven years. He has authored and co-authored over one hundred and fifty papers in the fields of electronic materials, epitaxial growth, and applications of focused ion and electron beams. He has also given over fifty invited presentations at national and international conferences in these fields. He is on the editorial board of several major journals, and has edited several proceedings and reference volumes. In 1997 he was the President of the Materials Research Society, the leading international society in the field of materials science and engineering.
TIME AND LOCATION: November 11 at David's Restaurant at the Santa Clara Tennis and Golf Club, at 5151 Stars & Stripes Drive in Santa Clara, CA (95054). This is just east of the Santa Clara Convention Center. Dinner choice of: London Broil or Seafood Brochette served with lemon butter sauce or a Vegetarian (Pasta Primavera) plate. Social at 6:00 p.m., 6:45 Dinner and 8:00 p.m. Talk
Cost: ASM/IEEE Members $16, Students $8 and Guests $18 (with an additional two dollars if no RSVP the Monday before the event (ie, Nov 9th ) Reservations: Brock Hinzmann 650-859-4350 or email IEEE Santa Clara Valley Chapter Reliability Society contact David Su (davidsu-at-aol.com). Please include choice of meal !
} The carbon tape is successful with distributing the effects of } charging but not heat. I, at least, can only suggest you sputter } with a heat conducting metal ... e.g., Al, Au, Cu ... most any metal } would help, but I realize it would superimpose spectra and degrade } your BSE contrast. Short of that, you'd need to lower the energy in } your beam with either lower beam currents or accel voltage. } I'll also be looking for other ideas ... hope this helps :o) } } cheerios, shAf } If heat is the problem, how about using a metal adhesive tape - you should be able to find adhesive-backed copper and aluminium tapes in the microscopy consumables catalogues. Turn into a loop, so it will stick to your stub. Ideally (?), bend over one corner, so you have metal to metal contact at some point between the tape and the stub to conduct heat and electricity. However, if the carbon tape is being damaged, where is the adhesive migrating too?
Keith
-- Dr. Keith R. Hallam University of Bristol, Interface Analysis Centre, Oldbury House, 121, St. Michael's Hill, Bristol, BS2 8BS, England Telephone: + 44 (0)117 925 5666 | E-mail: k.r.hallam-at-bristol.ac.uk Facsimile: + 44 (0)117 925 5646 | URL: http://zeus.bris.ac.uk/~phkrh/
We are looking to purchase a heating stage for our Philips FEG ESEM. We have information about the Philips stage which can reach 1000C but some workers will require still higher temperatures, and their 1500C stage is not on the market yet. Does anyone have experience of using heating stages and at what temperatures? How do different manufacturers compare and how easy are other stages to adapt to the ESEM. In particular will we still be able to use the Gaseous SE detector or are they designed for use with cooled conventional SE detectors? Any info will be much appreciated.
Cheers Nikki
Nikki Bock EM Technician Dept. Materials Engineering University of Nottingham Nottingham NG7 2RD (0115) 9513759/9513871 Email: emznjb-at-hermes.nottingham.ac.uk
Owen, Since you don't need much adhesive to tack down 10 micron particles, you may try applying a very thin layer of a fluid adhesive (such as Microstick) to a pyrolytic carbon planchet. This form of carbon has a flat, hard, glass-like surface. A drop of adhesive applied to the carbon may be spread very thin by drawing out with a cover slip. Since the organic adhesive layer is thin, you will probably get your required conductivity. There is no observable background structure. Planchets are about $35 each.
Dennis.
________________________________________________________ Dennis C. Ward voice: 202-324-2982 FBI fax: 202-324-4018 Microanalysis Laboratory e-mail: DCWard-at-concentric.net
It reads like a young someone looking for an easy way to get homework done.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supr. 10/21/98 8:22:11 AM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I just got a used Plasma Machine (International Plasma corporation). I do not know the age of it, but I have all the information.
Model: Reactor Center
S/N 11862-1
Number PM-1613
I would like to know if someone can help me to get the address of the manufacture or get the manual with the wiring diagram. Any help would be appreciated
} In previous email correspondence it was suggested by few people to use Evans } Blue stain as a counterstain on FITC labelled sections. Is there a special } protocol for it.
The advice to use Evans blue probably came from a research worker who had experimented intelligently and come up with something useful for his/her purposes. If you want to use this or any other dye as a counterstain for immunofluorescence, you will have to try and err. There is no such thing as a "protocol" for this sort of thing.
Evans blue and similar dyes (notably trypan blue) are not fluorescent but they become fluorescent when they bind to some (but not all) proteins. If you stain a section with 1% Evans or trypan blue at any reasonable pH (meaning 3 to 7), you get everything in a weary blue colour, with some feeble fluorescence here & there. The fluorescence can be seen (weakly) with rhodamine-type setups (green excitation; orange emission) or with broad-band UV and a colourless barrier filter (looks yellow). If there's too much blue dye that's not protein-bound, it quenches all the fluorescence, as do most blue dyes.
If you need a counterstain to show you where you are in an immunostained section, I'd recommend a weakly fluorescent basic dye, which will show nuclei, cytoplasm of RNA-rich cells, and acidic carbohydrates (cartilage matrix, some mucus, etc). Neutral red does this pretty well: 0.001 to 0.01%, at or near pH 4. By changing the filter block you can vary the relative prominences of this dye and various other fluorochromes. Try various concentrations of neutral red on any old sections to decide on an ideal time and concentration for this fluorochrome.
It is not possible for anyone to provide a "protocol" for fluorescent counterstaining. The technique varies with the requirements of the investigation.
John A. Kiernan, Department of Anatomy & Cell Biology, The University of Western Ontario, LONDON, Canada N6A 5C1
I am working with glass samples in both the SEM and TEM. We have access to FEG instruments and are contemplating a new FEG-SEM. Our current carbon coaters are dirty diffusion pumped systems. I know from experience that this equipment will cause contamination when a FEG instrument is used even when I use the LN2 trap for several hours before I open the system to put my sample in. I am currently using my BalTec RES 100 ion mill to sputter coat carbon on both sides of my TEM samples. (Yes, I know that I need only coat one side, but I have a specific reason for coating both sides.) However, this process is relatively slow and ties up my ion mill.
I know the arguments for having a sputter system for high resolution SEM imaging, but my interests are concerning carbon coating.
Here are my questions: What carbon coating systems are currently compatible with FEG microscopes? Is an turbo pumped evaporation system good enough or is a turbo pumped sputter deposition system required?
I would be interested in hearing from both users about their experiences and from vendors about what instruments they have and pricing information.
Thanks in advance.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Well, it's an interesting question, even if someone's just having a little fun with us. Another factor here is that the Earth is not a perfect sphere, it has a larger diameter if measured around its equator than North Pole to South Pole. So around the world, but using what as the path of travel?
} } The earth is a sphere. A quarter is a disk. If you're going to lay the } quarters on the surface of the sphere, the answer is very different. } ANyhow, do you think this merritted a serious answer or do you think } somebody is trolling with nasty bait? } } -------------------------------------------- } Michael Cammer } email sent from an account of the Analytical Imaging Facility } The Albert Einstein College of Medicine of Yeshiva University } 1300 Morris Park Ave. Bronx, NY 10461 } (718) 430-2890 FAX: (718) 430-8996 } http://www.ca.aecom.yu.edu/aif/ } -------------------------------------------- } } On Wed, 14 Oct 1998, Alwyn Eades wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I apologize for the tone but I feel that this was a terrible answer to send } } out. I assume that the person who asked the question is either young or } } not too familiar with the ideas of science. Therefore to imply that the } } magnification requested can be known so accurately (more accurately than we } } know any of the fundamental constants, for example) is leading the reader } } astray, to put it mildly. If the diameter of a quarter varies by, say, a } } part in a thousand from one to another, the answer would be 574 times ten } } to the power six i. e. 574 million. All the other figures are wrong. } } } } Alwyn Eades } } } } At 09:04 AM 10/14/98 -0400, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Earth Diameter = 12,756.28 KM -at- Equater } } } Quarter Diameter (US) = 7/8 inch } } } } } } The Earth is 573,960,854.9 times larger, so that's your mag. } } } } } Alwyn Eades } } Department of Materials Science and Engineering } } Lehigh University } } 5 East Packer Avenue } } Bethlehem } } Pennsylvannia 18015-3195 } } Phone 610 758 4231 } } Fax 610 758 4244 } } jae5-at-lehigh.edu } } } }
-- ****************************** Jim Haley Applications Engineer I-CUBE 2411 Crofton Lane, Suite 14A Crofton, MD 21114 voice: (301) 858-0505 fax: (301) 858-0615 web site: http://www.i-cubeinc.com e-mail: haley-at-i-cubeinc.com ******************************
I've used heating stages from Oxford instruments in the past up to 500C, and I believe they make other models for much higher temperatures (at least 1000C+ from memory). I'm sure that their apps people would be able to advise on the suitability in the ESEM.
Hope that helps,
Jeremy
} ---------- } From: NICOLA BOCK[SMTP:Nicola.Bock-at-nottingham.ac.uk] } Sent: 21 October 1998 10:23 } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM Heating stages } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Dear Microscopists } } We are looking to purchase a heating stage for our Philips FEG ESEM. } We have information about the Philips stage which can reach 1000C but } some workers will require still higher temperatures, and their 1500C } stage is not on the market yet. Does anyone have experience of using } heating stages and at what temperatures? How do different } manufacturers compare and how easy are other stages to adapt to the } ESEM. In particular will we still be able to use the Gaseous SE } detector or are they designed for use with cooled conventional SE } detectors? } Any info will be much appreciated. } } Cheers } Nikki } } } Nikki Bock } EM Technician } Dept. Materials Engineering } University of Nottingham } Nottingham NG7 2RD } (0115) 9513759/9513871 } Email: emznjb-at-hermes.nottingham.ac.uk }
One of my first encounters with this message many years ago had nothing to do with a program error. One of the six 1 MB SIMMs in my 386 (I told you it was a long time ago) started failing. Whenever I accessed that flaky memory location, the error would pop up. I tested it by opening multiple copies of a program (Word 2.0) and the failure would always come at the same place. I moved the SIMMs around in the slots and the problem moved around to. I isolated the flaky SIMM, replaced it, and the problem went away. BTW, the memory all passed the several memory test programs I ran it through.
I have also seen the problem develop when a cooling fan fails. The computer runs okay for a while, but when it starts getting taxed, it fails. Both problems lead to a corruption of the instructions in memory, so the program on disk may be fine, but it gets screwed up once it is loaded into memory.
These may not be the source of your problem, but it is probably worth a check.
Warren Straszheim
At 03:15 PM 10/21/98 +1000, you wrote: } } Hello everyone, } } I'm having a strange problem in saving high definition images onto the hard } disk. This has only recently occurred. Yesterday, the first high definition } image that I attempted to save took around 8 minutes, after which the system } froze on me preventing me to save any further high definition images. The } situation now is that I am only able to save standard definition images, but } not high definition images. When the system freezes there is a warning sign } that says, } } "Server Cause a General Protection Fault in Module SERVER.EXE at 0001:OD89" } } Rebooting the computer does not solve anything. Defragmenting the hard drive } did not help. } I use SEM Philips XL20 and software V 5.21. The computer is using Windows } 3.1 operating system and has an abundance of hard disk space. } } Any suggestions greatly appreciated. } } Khanh Tran } Deakin University } 662 Blackburn Road } CLAYTON, VIC. 3168 } AUSTRALIA
Hello Friends, Does anyone have a wiring diagram for the stage control (right side) for = the Hitachi H-600 TEM? One of our users unspooled and kinked the wire = that is attached to a push-pull, two spool, multi-pathway arrangement that = moves the grid. ( Looks like an old dental drill in minature! ) We do not = have a service contract and Hitachi is not sure that any of their = engineers in the Midwest has been trained to restring it. They may have = to dismantle the scope and send the arm out for re$tringing. I do a lot = of fishing...how hard could it be if a diagram is available. I am willing = to give it a try to get the scope operational in the least expensive = manner possible. Thanks. FAX 1-708-216-3913 Linda Fox Loyola University Medical School
Dear Nicola, I would suggest you try a company, such as Deben UK Ltd. (http://www.deben.co.uk), that make special stages for any microscope. They should be able to answer your questions. You wrote: } Dear Microscopists } } We are looking to purchase a heating stage for our Philips FEG ESEM. } We have information about the Philips stage which can reach 1000C but } some workers will require still higher temperatures, and their 1500C } stage is not on the market yet. Does anyone have experience of using } heating stages and at what temperatures? How do different } manufacturers compare and how easy are other stages to adapt to the } ESEM. In particular will we still be able to use the Gaseous SE } detector or are they designed for use with cooled conventional SE } detectors? } Any info will be much appreciated. } } Cheers } Nikki } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Microscopists } } We are looking to purchase a heating stage for our Philips FEG ESEM. } We have information about the Philips stage which can reach 1000C but } some workers will require still higher temperatures, and their 1500C } stage is not on the market yet. Does anyone have experience of using } heating stages and at what temperatures? How do different } manufacturers compare and how easy are other stages to adapt to the } ESEM. In particular will we still be able to use the Gaseous SE } detector or are they designed for use with cooled conventional SE } detectors? } Any info will be much appreciated
Hi Nikky,
I cannot answer your question directly as I have not used SEM hot stages but I do use TEM hot stages in a variety of different gasses and pressures. My hot stage uses a W heating element and will get to 1000C in vacuum. However, it will only get to 700C in 20mbar H or He before it burns out as it needs so much more power to overcome the heat loss in the gas. It will only hold 350(ish)C in oxygen before the W oxides finally burn the wire out, 300C is OK, for hours 400C for tens of minutes. Other gasses have similar problems. These problems could be improved or eliminated with better stage design or alternative materials. Check specification of the hot stages under the gas type and pressures that you are interested in working at.
Good luck, Ron. =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
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Owen, Since you don't need much adhesive to tack down 10 micron particles, you may try applying a very thin layer of a fluid adhesive (such as Microstick) to a pyrolytic carbon planchet. This form of carbon has a flat, hard, glass-like surface. A drop of adhesive applied to the carbon may be spread very thin by drawing out with a cover slip. Since the organic adhesive layer is thin, you will probably get your required conductivity. There is no observable background structure. Planchets are about $35 each.
Dennis.
________________________________________________________ Dennis C. Ward voice: 202-324-2982 FBI fax: 202-324-4018 Microanalysis Laboratory e-mail: DCWard-at-concentric.net
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Hallow, Thanks for the information about your new catalog. I shall very much appreciate a copy of the catalog XIII. Thanking you.
Your truly, C.L.Singla EM Lab, Department of Biology University of Victoria P.O.BOX 3020 Victoria, BC. Canada V8W 3N5
As it turns out, the individual looking into the problem here has had some contact with our suppliers re: the Y2K problem. They have all been extremely helpful and there are many resources available. I hope that I did not infer otherwise with my original post.
I have attached some of the information as submitted from our equipment suppliers and manufacturers including Joel USA, Soquelec Ltd., Oxford Instruments America Inc. and Nissei Sangyo Canada Inc..
I hope it is helpful to others in the group as well.
Thanks for all the input.
Wayne England
_______________________________________________
I saw your message about Year 2000 compliance on the Listserver this morning. If you would send me your address, I will be glad to mail you the statement covering all the Oxford microanalysis systems (including the eXL).
Thanks,
Graham Bird Business Manager Oxford Instruments America Inc. Microanalysis Group 130A Baker Ave Extension Concord MA 01742 USA
Please note that JEOL does have a Y2K statement on our Homepage at http://www.jeol.com along with two contacts for more information, one of whom is a Director and the other who is devoting full time to this problem. This person (corey-at-jeol.com) can give you the current status of any JEOL instrument.
Regards Steve Hamilton Marketing Manager JEOL USA, Inc. hamilton-at-jeol.com
Analytical Imaging Facility wrote: } } The earth is a sphere. A quarter is a disk. If you're going to lay the quarters on the surface of the sphere, the answer is very different. } ANyhow, do you think this merritted a serious answer or do you think } somebody is trolling with nasty bait?
} -------------------------------------------- } Michael Cammer } email sent from an account of the Analytical Imaging Facility } The Albert Einstein College of Medicine of Yeshiva University } 1300 Morris Park Ave. Bronx, NY 10461 } (718) 430-2890 FAX: (718) 430-8996 } http://www.ca.aecom.yu.edu/aif/ } --------------------------------------------
Michael:
Are you serious? What kind of "nasty bait' could this be? Let's drop this thread.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
I'm working with a student who is trying to determine the degree of homogeneity in tool steel using microprobe. She is short of good references for the fundamental statistical equations involving x-ray measurements in electron beam instruments. Can anyone provide a few relevant references? Thanks in advance.
Owen
============================= Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
Would anyone out there have a contact number for Linkam Company which makes heating/cooling stages for light microscopes or a contact number for a distributorship of Linkam products in the New England area? Thanks for any help. Don Gantz Boston Univ School of Medicine gantz-at-med-biophd.bu.edu
We have a few items that we'd like to sell. One is a Balzers Freeze Etch Apparatus. The specimen chamber on this system is glass (model # anyone?), and has a Freeze Etching Device Control unit GA-1, with a Power Control Unit BSV 202, and a Cold Cathode Gauge PKG 010 Pirani. This system has sat idle for a number of years now. Also, we have a few MT-2 mictomes for sale. Interested parties can email me or reach me by phone at 319-335-8142, fax 319-335-6710. Thanks, Randy -- Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
} -----Original Message----- } From: Owen P. Mills [mailto:opmills-at-mtu.edu] } Sent: Wednesday, October 21, 1998 10:36 AM } To: MICROPROBE-at-ftp.microanalysis.org; Microscopy-at-Sparc5.Microscopy.Com } Subject: materials - homogeneity statistics } } } Hello: } } I'm working with a student who is trying to determine the degree of } homogeneity in tool steel using microprobe. She is short of good } references for the fundamental statistical equations involving x-ray } measurements in electron beam instruments. Can anyone provide a few } relevant references? Thanks in advance. } } ...
I have a student's thesis ... somewhat dated ... she synthesized reference standard glasses for empirically determining EPMA alpha factors. Anyway, there are several pages dedicated to her quantification of the glasses' homogeneity, calculated with respect to counting statistics and what would be expected from the Chi-square distribution. Of anyone is interested, contact me directly ... subject: "chi-square homogeneiety" ... I'll try to put it thru OCR and clean it up ...
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
We have proposed to label a a molecule that is usually sequestered to the inner leaflet of the cytoplasmic membrane using a protein specific for that molecule. We proposed to use colloidal gold with TEM to determine which side of the membrane the molecule is located. We have been told that we will not be able to do this study because the gold-protein complex is so large compared to the width of the bilayer membrane that we could not possibly determine which side of the bilayer the label is on.
Can anyone supply me with a reference to any study that shows that TEM can be used to determine which side of a membrane bilayer a specific marker is located? It does not have to be specifically colloidal gold labeling any method would be fine.
Thank you
Please reply by personal e-mail wiessner-at-mcw.edu
Dear Chuck, } } Dennis has a good idea. I would substitute freshly-cleaved mica for } the carbon planchet...its a lot cheaper. } Since the idea is to do microanalysis, having a substrate of sim- ilar composition to the specimen would not be advisable. Yours, Bill Tivol
The estimation of whether or not a specimen has a uniform composition, or is 'homogeneous', usually requires the application of some statistical method. This is a subject which received a lot of attention in the days when microprobe methods were being developed, but which seems to be little mentioned these days.
There is a discussion of the basic concepts involved and the methods commonly used in describing the level of specimen homogeniety in Sect. 8.6 of the book 'Scanning Electron Microscopy & X-ray Microanalysis' by Goldstein, et. al. Plenum Press, that will probably meet your student's needs
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
It was announced Oct 1 that manufacturing of the Kevex Sigma EDS line will be transfered to Noran. The XRF line is going to Spectrace Instruments. The fate of the Kevex service group is uncertain.
A long competive battle ends and a proud flag falls.
I have experienced difficulty in finding a supplier for the large light bulb, Thorn 110-120 V, 200W, Opale E27, P3/15 which is used in my Durst Laborator 138S condenser enlarger. If anyone has information regarding this, I would appreciate hearing from you.
Mary North Electron Microscopy Lab Loma Linda University Medical Center Loma Linda, CA northstar44-at-juno.com
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com or call Juno at (800) 654-JUNO [654-5866]
Hard to know where to begin with such poor resolution. I assume that the operating setup hasn't changed, i.e. you are using the same accelerating voltage, condensor settings, working distance, etc., as before and the resolution has changed. Of the above, the condensor will have the most marked effect on resolution - the higher the current the better the resolution and the smaller the beam current. As I recall, the S120 doesn't have a condensor current readout, so it's conceivable the condensor circuit is malfunctioning. It should deliver 2 - 3 amps or so for much better resolution than you are getting.
Proper filament alignment within the cathode, properly cleaned and well aligned optical parts would be my next check. The closer to the beam something is contaminated, the greater the effect. It takes very slight, invisible to the eye, contamination on the hole in an apeture to have disastrous effects. How are you cleaning the apetures? Heating to dark cherry red in a vacuum evaporator is the best for any apeture. Open air flaming is good for platinum apetures. If you are cleaning them by hand with metal polish, make sure that you use an ultrasonic cleaner after first cleaning off the polish by hand.
BTW, many people use acetone to clean such parts in an ultrasonic cleaner. I quit doing that years ago after a customer, an ex-NASA engineer, informed me that acetone doesn't cavitate the way most fluids do. He had to research it for them, so I've trusted his opinion and use ethyl alchohol. While acetone does leave a residue, it doesn't present a problem and I often use it as a final rinse or at least to manually clean the parts before ultrasonicing.
If the change in resolution was sudden, the condensor works properly, and the column appears to be clean, I'd take a real close look for a fiber caught somewhere in the column. Take everything apart and blow everything off well with filtered air, dry nitrogen or canned air while carefully re-assembling to prevent contamination of assembled parts.
Finally, can you give any more clues? Any change in manufacturers for filaments or apetures? Is the limitation a huge astigmatism? Any problem getting a filament image that is well centered? Are there any indications of large electromagnetic or vibration problems (saw-toothed edges at fast scan rates)?
} Dear Friends, } } I have a problem in resolution of Scanning Electron Microscope S120, } Cambridge Instrument. I have tried cleaning the SEM column, as I do } everytime when there is a astigmatic image. But, recently I am } unable to get good image at even as low as 1000X. What might but } the problem? } } Thanking you all in advance. } } Yours faithfully, } } Rajdeep Dongre } Electron Microscopy Laboratory } Agharkar Research Institute } G.G. Agarkar Road, Pune - 411004, } India } } E-mail : rajdeep-at-aripune.ernet.in } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
I recently bought a Kodak's 120 MDS system for taking digital photographs, but I'm having some problems to which I have not found a solution yet: a bright, red, somewhat diffuse, spot appears on center of every picture. The MDS software has a specific function to solve this problem, but this doesn't helps too much. Kodak manual also recommends to reduce as much as possible the light intensity, but the spot even appears. Now, I process the pictures using the photopaint's artifacts, but it takes too much time and the product is not as good as one would expect.
We just published the following paper in which you can find references concerning zircon dating (suzuki et al).
Cocherie, A., Legendre, O., Peucat, J.J., and Kouamelan, A. Geochronology of polygenic monazites constrained by in situ electron microprobe Th-U-Total Pb determination: implication for Pb behaviour in monazite. Geochimica et Cosmochimica Acta 62:2475-2497, 1998.=20
} ---------- } De : Joil Jose Celino[SMTP:jjc0704-at-mailexcite.com] } Date=A0: jeudi 22 octobre 1998 14:34 } A : MICROPROBE-at-ftp.microanalysis.org; } Microscopy-at-Sparc5.Microscopy.Com; Owen P. Mills } Objet : Isochron ages of monazites and zircons } =20 } Hello all, } =20 } I'm working with some granites in mobile belt of Brazil with ~ 800 = Ga. } Does anybody know about dating zircon with electron microprobe?? } =20 } Thanks } --- } Joil Jos=E9 Celino } University of Brasilia - UNB } Institute of Geoscience } Brasilia - BRAZIL } Fone (061) 321-0410 C=F3digo: 6181948 } =20 } =20 } =20 } Free web-based email, Forever, From anywhere! } http://www.mailexcite.com } =20
Try a company in California at 1-800-8Lights, they can get hard to get bulbs.
Regards,
Michael Coviello Materials Science UT Arlington
UT Arlington Attn: Michael Coviello Box 19031, 500 W. 1st Street Arlington, TX 76019
dAt 10:19 PM 10/21/98 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Try calling Bulb Direct, Pittsford, NY, (800) 772-5267, www.bulbdirect.com ======================= } I have experienced difficulty in finding a supplier for the large } light bulb, Thorn 110-120 V, 200W, Opale E27, P3/15 } which is used in my Durst Laborator 138S condenser enlarger. } If anyone has information regarding this, I would appreciate } hearing from you. } } Mary North } Electron Microscopy Lab } Loma Linda University Medical Center } Loma Linda, CA } northstar44-at-juno.com
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
Regarding text recommendations, Scanning Electron Microscopy--A Students Handbook by Postek, Howard, Johnson, and McMichael (Ladd Research Industries) is an excellent text book for college level students. Regarding materials/subject matter, you might want to consider including specific application lectures such as SEM in the microchip industry, SEM in medicine, SEM in forensics, etc. If you want to contact me directly, I can give you some forensic pointers. Also, Dr. Darlene Southworth of the science department of Southern Oregon University in Ashland Oregon has taught a very popular one semester undergrad course on SEM for years. She could be a good source of information/guidance for you.
(Netscape Messaging Server 3.5) with ESMTP id AAA379C for {Microscopy-at-Sparc5.Microscopy.com} ; Thu, 22 Oct 1998 09:03:33 -0700 Message-ID: {36300010.F1CC2BF8-at-nctimes.net}
What centrifuge do you recommend for concentrating virus specimens, like intestional for parvo? We only have $15,000 to spend. Does anyone use a Beckman Allegro 64R? It reaches 64g and I'm told it can bring down bacteriophages in 4 hours. We would really like to have a Beckman 55.2 Ti rotor, but we can't afford the centrifuge. Thank you! Cindy Shannon
I've got someone looking into the possibility of preparing samples of tool material that consists of diamond particles embedded in a cobalt matrix. Any ideas? How will I cut, grind and polish it?
Thanks,
Robin Griffin UAB Materials and Mechanical Engineering
My question: What metallographic mounting materials are safe for an SEM? Some of our users would like us to forbid usage of any mounting compound other than those that require both pressure and heat such as bakelite and its near relatives. They feel that the resolution of the machine will gradually degrade if we allow the usage of cold mounting materials. We have currently allowing the usage of some of the more stable (epoxy from leco and buehler for example) cold mounts. We don't see any change in vacuum when the beam strikes the sample mount but are we slowly degrading the image resolution by gunking up the column? To date the resolution still is fine but what about the future?!
Ronald Vane wrote: } } It was announced Oct 1 that manufacturing of the Kevex Sigma EDS line } will be transfered to Noran. The XRF line is going to Spectrace } Instruments. The fate of the Kevex service group is uncertain. } } A long competive battle ends and a proud flag falls. } } See www.kevex.com for more details. } } Ronald Vane } XEI Scientific --
Actually, the service is being transferred here (to Middleton, WI) as well. All the California service number(s) and 800 number(s) should remain the same, so that for all practical purposes "Kevex" (at least the customer service/support) will continue to exist, and both of our product lines may in fact be strengthened by the experience. Please let me add that not a few here at NORAN share your regret; we take no delight in cannibalizing our long-time soul mate, and mourn the loss of our sister. (Many perhaps didn't realize that Kevex was also a ThermoSpectra company for the last few years, so we haven't really even been competing for some time.)
-- Tim
--------------------------------------------------- Timothy G. Moeller | NORAN Instruments Inc., Sr. Software Engineer | a ThermoSpectra company {tmoeller-at-noran.com} | {www.noran.com} ---------------------------------------------------
} -----Original Message----- } From: "grial-at-fibertel.com.ar"-at-sparc5.microscopy.com } [mailto:"grial-at-fibertel.com.ar"-at-sparc5.microscopy.com] } Sent: Sunday, May 24, 1998 8:19 PM } To: microscopy-at-sparc5.microscopy.com } Subject: LM-problems taking digital photography } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I recently bought a Kodak's 120 MDS system for taking digital } photographs, but I'm having some problems to which I have not found a } solution yet: a bright, red, somewhat diffuse, spot appears on center } of every picture........ } } I would really appreciate any suggestion! } } Thany you in advance! } } Carlos E. Barbosa } } At the MM98 meeting in Atlanta there was a short course on Practical Digital Imaging where the instructor, John Mackenzie jr., in his praise for this camera also pointed out that in some microscopes this red spot will occur and can not be removed. He did not mention the names of these microscopes but did know of brands that did experience this phenomenon.
Sam. O. Mancuso Group Leader, Physical-Mechanical Metallurgy Special Metals Corporation 4317 Middle Settlement Road New Hartford, New York 13413 U.S.A. {} {} {} Phone: (315) 798-2920 Fax: (315) 798-2001 email: mancuso4-at-ix.netcom.com}
In years past we had ugly problems when cold mounting materials were used to mount specimens for examination in our SEMs (as discussed in some detail in Sect. 2.10.4d. p. 74, of my book on 'Vacuum Methods in Electron Microscopy'; see:sales-at-portlandpress.co.uk, or http://www.cccbi.chester.pa.us/spi/catalog/books/book48.html). Eventually, we found that we could reduce the problem to an acceptable level by requiring that the mounting materials be measured with great care to ensure the proper proportions of resin and hardener, and that the mounted specimens be allowed allowed to stand for at least 24 hours to ensure complete curing, and then be prepumped for 24 hours in a chamber such as we used to use for prepumping film. It would also be valuable to heat the chamber (wrap a heating tape around it) to as high a temperature as the specimens would stand during pumpout.
We never experienced as much trouble with thermosetting materials such as the Bakelite, diallyl phthalate and epoxy resins which are cured under heat and pressure, but still required a 24-hour pumpout for them too.
As also discussed in VM in EM, similar problems can arise from suspensions of polishing abrasives, etchants, and conductive paints which are also commonly used in preparing and mounting specimens.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
} My question: } What metallographic mounting materials are safe for an } SEM? Some of our users would like us to forbid usage of any } mounting compound other than those that require both } pressure and heat such as bakelite and its near relatives. . . . . . . . } Robin Griffin }
We have been using Struer's EPOFIX epoxy mounts for years in our SEM with no noticeable degradation in either vacuum or image and x-ray resolution when prepared in proper porportions. In fact because the margin around the specimen is tighter with the epoxy than the bakelite, there is less out-gas due to entrapped water and solvents from cleaning procedures.
Sam. O. Mancuso Group Leader, Physical-Mechanical Metallurgy Special Metals Corporation 4317 Middle Settlement Road New Hartford, New York 13413 U.S.A. {} {} {} Phone: (315) 798-2920 Fax: (315) 798-2001 email: mancuso4-at-ix.netcom.com
In our research we study growing polymer particles under an ordinary optical
microscope at 4-20 times magnification. The polymer particles lie on a black
surface to assure a good contrast between the particles and the background. Good contrast is important because the size of the particles is determined with software, based on differences in light intensity. Of course, a digital camera (pieper FK7512-IQ) is connected to the microscope.
Our problem is that at the beginning of the experiment the background has some noise; after some time (when the particles are much larger, so more white light is on the picture) this noise decreases and almost disappears. This noise in the beginning of the experiment isn't very severe and the software can still determine the size of the particles. However, the fading away of the background noise has some influence on the determination of the size of the particles: during the fading the measured growth decreases (owing to the fading).
Does somebody experienced such a problem before, knows what causes it, knows
a solution or knows how to control the fading. We think it might be some hardware problem or an automatic adjustment of some kind of diaphragma (the camera sees more light, possible it automatically adjusts for that).
We hope someone can help us with this problem!
Thanks in advance for any help or suggestions you may have.
Ronald Capel
University of Twente Faculty of Chemical Technology PO-Box 217, 7500 AE, Enschede, Netherlands r.capel-at-student.utwente.nl
} } On Thursday, October 22, 1998 8:34 AM, Joil Jose Celino } [SMTP:jjc0704-at-mailexcite.com] wrote: } } Hello all, } } } } I'm working with some granites in mobile belt of Brazil with ~ 800 Ga. } } Does anybody know about dating zircon with electron microprobe?? } } } } Thanks } } --- } } Joil Jose Celino } } University of Brasilia - UNB } } Institute of Geoscience } } Brasilia - BRAZIL } } Fone (061) 321-0410 Codigo: 6181948 } } } } } } } Check out the paper by Cocherie et al that just came out in Geochimica et } Cosmochimica Acta (v. 62, p.2475, 1998), "Geochronology of polygenetic } monazites constrained by in situ electron microprobe TH-U-total lead } determination: Implication sfor lead behaviour in monazite". An interesting } application. } } Jim McGee } } {} {} {} {} {} {} {} {} {} {} {} {} {} } James J. McGee (jmcgee-at-sc.edu) } Department of Geological Sciences } University of South Carolina } Columbia, SC 29208 } } TEL: (803) 777-6300 FAX: (803) 777-6610 } } }
Dear friends, This may be a very basic question. We use formvar film strengthened with a thin layer of carbon as the literature did. My question is if a thin vaporated gold layer can replace the carbon layer to strengthen the formvar film? Is it better to use gold than carbon? Any advantages/disadvantages for them? Thanks in advance for your assistance.
BTW, is there a FAQ file of this mailing list? I am afraid to repeat the issue discussing before.
I have a series of tiff secondary electron images of serial sections through a polished mineral grain. Could someone please advise me as to the best software available for doing 3-D reconstructions from these tiff files on my MAC (266mHz G3). Each is around 1 meg and they are about 60 sequential images.
Thank you
Brendan
Brendan J. Griffin Centre for Microscopy and Microanalysis The University of Western Australia Nedlands, WA, AUSTRALIA 6907 ph 61-8-9380-2739 fax 61-8-9380-1087
Our TEM, a JEOL 2010 is up for a service contract. In the opinion of the list, how often should such an instrument be serviced (assuming regular use)? For example, one small visit for a general check and another for a full service, pole pieces cleaned, emission chamber etc.
What are the general terms for service contracts around the world.
Many thanks
Keith.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Your enquiry interested me. Sorry for my late reply, but have not been in the office for a while. Freeze-fracturing worked very well for a study I conducted on cement paste using a SEM with a cryogenic stage. This work was done with Dane Gerneke at the University of Cape Town, South Africa. Essentially, one freezes the cement paste specimen (which of course contains water bonded and non- bonded) in liquid nitrogen and then in an evacuated sample preparation chamber the specimen is fractured. The specimen was then transferred in vacuo to the SEM stage. The beauty of this was that the fracture surface that we studied is a different type of surface than when it is done dry. The freeze-fracturing actually broke through the cement grains as opposed to round them, the latter which occured under normal room temperature conditions.
Sincerely, Quirina
\|||/ (o o) *-----------------oOO--(_)--OOo
Quirina I. Roode-Gutzmer PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-------------------------------Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ .oooO-------------------------* ( ) \ ( \_)
THE 1500 degree heating stage is in productionn. Jo Long and TRisha RICE of Philips indicated that there are 4 or 5 in use already. FURTHer, another 1500 degree stage was being installed today in their demo lab in Mass. Give your Philips rep another call.
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Dear Nicola, I would suggest you try a company, such as Deben UK Ltd. (http://www.deben.co.uk), that make special stages for any microscope. They should be able to answer your questions. You wrote: } Dear Microscopists } } We are looking to purchase a heating stage for our Philips FEG ESEM. } We have information about the Philips stage which can reach 1000C but } some workers will require still higher temperatures, and their 1500C } stage is not on the market yet. Does anyone have experience of using } heating stages and at what temperatures? How do different } manufacturers compare and how easy are other stages to adapt to the } ESEM. In particular will we still be able to use the Gaseous SE } detector or are they designed for use with cooled conventional SE } detectors? } Any info will be much appreciated. } } Cheers } Nikki } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
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Hello all, I would thank you for sending me suggestions/references about anhydrous fixation methods for light microscopy, for plant material= (leaves). This method should permit further histochemical staining (like Feulgen reaction, P.A.S. reaction, use of Sudan Black, and others). The need for an anhydrous fixation method cames because I study desiccated material very reactive to water, so that any drop of water (in a common buffer, for example) could potentially change the results. I tought to use a modification of Hallan's (1976) anhydrous fixation for TEM (with DMSO instead of water) but I do not know if this method permits the further use of histochemical stainings. It would also be very usefull if someone could indicate me a list for cytoquemistry/histochemistry and/or for light microscopy discussion. As this may be not a subject of general interest to this list, I would suggest the people who could help me to send the messages direct to= me. Thank you in advance, Estela ************************************ Dr Estela S. Rossetto Universidade S=E3o Francisco F.F.C.L. Av. S=E3o Francisco de Assis, 218 12900-000 Bragan=E7a Paulista - S.P. Brasil e-mail: esrossetto-at-lexxa.com.br ************************************ =20 =20
Why not grab an image with no specimen, and use that image in a background subtract? I rountinely do this to get rid of uneven illumination and small imperfections (dirt) in the light path. The only real difficulty is that you need to collect a background image for each session, objective, illumination regime, etc., because the nature of the defect changes with any adjustment. But it works for me.... --
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Dear all, does anybody have experience with immunocytochemistry on Ni-, Au= - grids with a temperature probe controlled laboratory microwave oven? Is it also time co= nsuming and one could achieve better or comparable staining results protocols? I think, on= e problem could be the irradiation of the metallic grids, leading to overheating spots on the gri= d bars and this may precipitate the antibodies which would cause useless results. Is this neg= ligible? Many thanks for any suggestions Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=E4tsstrasse 25 Germany 33615 Bielefeld phone: 0521 1065592 fax: 0521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
We have two PMs a year for our 2010 (they are both part of the service contract). In general ,the pole pieces should not be touched, but the condenser and objective apertures might need to be cleaned/replaced depending on how bad they are. Also, the OBJ. aperture mechanism in our scope needs an occasional lubrication. This is done during the PM. During the PMs the rotary pump oil is replaced , they do a leak test for the gun and they check some of the electronics. They also fine tune some of the alignment ( in DADJ ^1).
I hope this helps
Jordi Marti ---------- } From: Keith Moulding To: Microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
I have also had good luck with epofix as well as hot pressed (thermoset) resins. As stated by Bigelow, it is very important to mix and cure properly to avoid outgasing.
Certainly not all resins are created equal. I forget the name (mental block?), but a few years ago we experimented with a bubble gum pink mount. It took a month at 120F in a vacuum oven before I could reach 1x10-3 torr! Heard it was discontinued - Thank goodness. Could smell the mount... If you can smell it, forget it:)
Woody White McDermott Technology, Inc -----------------------------------------------------------------
My question: What metallographic mounting materials are safe for an SEM? Some of our users would like us to forbid usage of any mounting compound other than those that require both pressure and heat such as bakelite and its near relatives.
They feel that the resolution of the machine will gradually degrade if we allow the usage of cold mounting materials. We have currently allowing the usage of some of the more stable (epoxy from leco and buehler for example) cold mounts. We don't see any change in vacuum when the beam strikes the sample mount but are we slowly degrading the image resolution by gunking up the column? To date the resolution still is fine but what about the future?!
Is anyone here using EDX to measure the amount and depth of decarburization of steel? I'd like to analyze the depth of the decarburization of steel that has been through a furnace. I'd like to take a cross section of the slug and beginning at the outer edge, perform a semiquant analysis with my EDX at succeeding depths from the outer edge into the core, measuring for carbon content. Is anyone here using an SEM for this type of work? I'd like to know how you go about it and what the success and confidence level is.
Please note that as of September 1998 MAS, Inc. of Suwanee GA, USA will be providing service and parts for the Topcon 002B TEM. Please call 800-421-8451 for service or parts. The service contact is Art McCanna at the Suwanne, GA office. The address is listed below.
my collegue and I thank all vendors and friends from this listservers for all the comments, quotations, etc. related to my question about sputter/coater devices.
We are processing the information received which is of very much help for taking a final decision.
Silvia Montoro Centro Regional de Investigacion y Desarrollo Santa Fe Argentina
I have an ISI DS-130, about 15 years old, that lately has been getting streaks of normal scan and darker scan at 100X. I thought it might have been due to the filament getting old (230 hours) so I changed it. I guess it wasn't that because the streaks are back. They only appear at 100X and then tend to be across the upper portion of the picture, image file or screen. I don't think it's charging because that shows up as white lines. Does anybody out there have an idea as to what might be causing these lines? Any suggestions will be greatly appreciated. My fly guys are ready to fly off the handle, and my plant people are getting ready to leaf me.
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
We don't like formvar for our purposes here, preferring either continuous or holey carbon films. I think that evaporated gold would be disadvatageous for two reasons: additional diffraction contrast and mass. If you need a support film that avoids carbon, you might be better served by SiO films. } } } Dear friends, } This may be a very basic question. We use formvar film strengthened with a } thin layer of carbon as the literature did. My question is if a thin } vaporated gold layer can replace the carbon layer to strengthen the } formvar film? Is it better to use gold than carbon? Any } advantages/disadvantages for them? } Thanks in advance for your assistance. } } BTW, is there a FAQ file of this mailing list? I am afraid to repeat the } issue discussing before.
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
My I first state that I'm an employee of VayTek, Inc.
We sell VoxBlast; a 3D Volume Visualisation & Measurement software.
VoxBlast reads serial TIFF images and creates a volume. You can then apply a wide range of parameters and functions to diplay the image on screen. VoxBlast is available for the PowerMac, Windows, and UNIX platforms.
If you have any additional questions please feel free to contact us directly or for more immediate information then please visit our web page at www.vaytek.com where you can download a free demo version of VoxBlast.
Regards,
Chris MacLean, Ph.D. VayTek, Inc. 305 West. Lowe, Suite 109 P.O. Box 732 Fairfield, Iowa, 52556-0732
Dear Robin, I have successfully prepared this material by mounting as usual in cold-cure epoxy, grinding on coarse diamond, about 40 micron, then on 15 micron diamond, then finish the polish on 5 micron diamond. The polish isn't perfect, but it doesn't need to be for microanalysis. The sample was cut for me. You wrote: } } I've got someone looking into the possibility of preparing samples of tool } material that consists of diamond particles embedded in a cobalt matrix. } Any ideas? How will I cut, grind and polish it? } } Thanks, } } Robin Griffin } UAB Materials and Mechanical Engineering } } Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
We have full service contracts on all 5 of the TEMs here. The service contracts include a visit every 6 months during which the engineers check the operating parameters, change the pump oils, examine the apertures,etc: mostly routine stuff. The contracts also cover almost all parts, labor, and travel expenses.
The expense of the contracts is questioned on occasion by the administration. I have been able to show that the expense of the contracts have not been more than we would have paid for repairs without the contracts over the course of several years. } } Hi, } } Our TEM, a JEOL 2010 is up for a service contract. In the opinion of the } list, how often should such an instrument be serviced (assuming regular } use)? For example, one small visit for a general check and another for a } full service, pole pieces cleaned, emission chamber etc. } } What are the general terms for service contracts around the world. } } } Many thanks } } Keith. } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Dr. K. Moulding. } } Materials Characterisation and Preparation Facility } Hong Kong University of Science and Technology, } Clear Water Bay, } Kowloon, } Hong Kong. } } FAX: (852) 2358 2451 } TEL: (852) 2358 8724 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
Dear List members the following is presented in response to a recent question:
--------------------------------------
} Dear all, does anybody have experience with immunocytochemistry on Ni-, Au- grids with a
} temperature probe controlled laboratory microwave oven? I think, one problem could be the
} irradiation of the metallic grids, leading to overheating spots on the grid bars and this may
} precipitate the antibodies which would cause useless results. Is this negligible?
} Bernward Laube
----------------------------------------
NOTE: There is no localized heating within metallic specimen grids by microwave irradiation. The grids are much smaller than the quarter wavelength of a microwave (quarter wavelength in air is ~3 cm for a standard microwave oven frequency of 2.45 GHz). {fontfamily} {param} Geneva {/param} (Kok LP, Boon ME: Physics of microwave technology in histochemistry. Histochem J 1990, 22:381-388)
{/fontfamily} R {fontfamily} {param} Geneva {/param} eferences on microwave -accelerated immunocytochemistry techniques
1. Login GR, Dvorak AM: Microwave fixation provides excellent preservation of tissue, cells and antigens for light and electron microscopy. Histochem J 1988, 20:373-387
2. McQuaid S, Isserte S, Allan GM, Taylor MJ, Allen IV, Cosby SL: Use of immunocytochemistry and biotinylated in situ hybridisation for detecting measles virus in the central nervous system. J Clin Pathol 1990, 43:324-333
3. Ohtani H, Maruyama I, Yonezawa S: Ultrastructural immunolocalization of thrombomodulin in human placenta with microwave fixation. Acta Histochem Cytochem 1989, 22:393-395
4. Wouterlood FG, Boon ME, Kok LP: Immunocytochemistry on free-floating sections of rat brain using microwave irradiation during the incubation in the primary antiserum: light and electron microscopy. J Neurosci Methods 1990, 35:133-145
5. Haruna N, Monden T, Morimoto H, Murotani M, Yagyu T, Nagaoka H, Shimano T, Mori T: Use of a rapid microwave fixation technique for immunocytochemical demonstration of tumor necrosis factor, interleuken-1 a, and interleuken-1b in activated human peripheral mononuclear cells. Acta Histochem Cytochem 1990, 23:563-572
6. Login GR, Galli SJ, Dvorak AM: Immunocytochemical localization of histamine in secretory granules of rat peritoneal mast cells with conventional or rapid microwave fixation and an ultrastructural post-embedding immunogold technique. J Histochem Cytochem 1992, 40:1247-1256
8. Cuevas EC, Bateman AC, Wilkins BS, Johnson PA, Williams JH, Lee AH, Jones DB, Wright DH: Microwave antigen retrieval in immunocytochemistry: a study of 80 antibodies. J Clin Pathol 1994, 47:448-452
9. Gu J, Forte M, Hance H, Carson N, Xenachis C, Rufner R: Microwave fixation, antigen retrieval and accelerated immunocytochemistry. Cell Vision 1994, 1:76-77
11. Johnston PW, King G, Herriot R: Increased range of immunocytochemical markers in a case of acute lymphoblastic leukemia by microwave pretreatment of paraffin sections (Meeting Abstract). J Pathol 1994, 173:200A 1994
TWIMC, The thin layer of carbon on a carbon-formvar film is there to provide conductivity, not strength. A thin carbon film is not self-supporting, and the formvar is used to strengthen the carbon, not the other way around. Heat- ing and/or charging effects will break formvar films which have not been C- coated, so in that sense, C-coating will strengthen formvar, but it is not a mechanical strengthening. Au is, indeed, a good conductor, so it would provide the same charge and heat dissipating properties as C, but, since it scatters electrons very strongly, it would interfere with imaging--especially as evaporated Au forms islands giving a mottled appearance. Yours, Bill Tivol
740206-at-ucl.itri.org.tw-at-Sparc5.Microscopy.Com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Dear friends, } This may be a very basic question. We use formvar film strengthened with a } thin layer of carbon as the literature did. My question is if a thin } vaporated gold layer can replace the carbon layer to strengthen the } formvar film? Is it better to use gold than carbon? Any } advantages/disadvantages for them? } Thanks in advance for your assistance. } } BTW, is there a FAQ file of this mailing list? I am afraid to repeat the } issue discussing before.
We have in the past produced formvar substrates for customers who requested a gold coating. Gold, like carbon, will strengthen formvar. We do not know which is better.
I assume it depends on your application. Gold coating is more expensive. In a sitation where you can not use a carbon coating, gold or some other material could be substituted.
Disclaimer: Ladd Research is a commercial vendor of microscopy supplies and accessories.
John Arnott --
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
If anyone out there has done work with Malaria, and knows of the best TEM fixative for this little RBC parasite, please contact me at: lamiller-at-uiuc.edu
Carbon is favored over gold because it is less dense and is actually stronger. Gold (or platinum or palladium, etc) form a dense, obtrusive background and are much less supportive than carbon. That is the reason that carbon is being used in high tech carbon composites rather than other metals (strength and lightness). In addition, if one does x-ray analysis, the heavier metals generate too many spectral lines that may interfere with your analysis.
} This may be a very basic question. We use formvar film strengthened with a } thin layer of carbon as the literature did. My question is if a thin } vaporated gold layer can replace the carbon layer to strengthen the } formvar film? Is it better to use gold than carbon? Any } advantages/disadvantages for them? } Thanks in advance for your assistance.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I recommend to start working slowly. Why don't you get yourself the FREE software NIH IMAGE. It is not only free, but it is the standard.
http://rsb.info.nih.gov
Look out, also, for the also free modification called OBJECT IMAGE, which is even better. Also on one of those servers. Just don't forget to ASSIGN ENOUGH MEMORY to those programs before you start digging around in your huge files.
They can be both used to reslice the 3D block (then called stack). Object Image also can interactively BROWSE through your block in three planes at once in real time. This gives you a pretty cool idea of what things there might be to visualize.
Don't spend too much money before you know what you need, get organized with the 3D possibilites, and more important, the diagnostic relevance or irrelevance of some features, and look out for the capabilities of IDL, the BEST interactive graphics tool around (Interactive Data Language, Research Systems, www.rsinc.com).
This is independent advice. - Cheers Wolf
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:::::::: wolf schweitzer, md bern, switzerland :::::::: www.access.ch/private-users/wschweitzer/cyberpage.html
Thanks to all who responded to my posting. All the information has been helpful. Responses are summarized below.
David Rose W.L. Gore and Associates
===========================================
I've seen posts which responded to the original "HP vs Epson vs Alps" which imply Alps is an ink jet. The original poster may have been referring to the Alps 1300 which is a dual mode 600dpi ink jet and dye-sub printer for ~$500. I was personally impressed with the example Alps sent me, but I also hear it may be the slowest dye-sub on the market ... and we all know printer performance has more to do with how well the printer interfaces with the OS. I know nothing beyond this ... but certainly am curious if someone has practical experience.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
___________________________________________
Only real photo quality is dye sublimation (kodak, codonics, tektronics, etc) which are expensive (maybe $5K and up). Inkjets like you name, with special, expensive glossy paper and 1200 dpi are almost as good for a few hundred $. I like the Epson Stylus Photo (EX) myself. All will be color (but make good B&W) unless you get a laser printer. Dave Pevear, Houston
I can't tell you anything about Alps, but I do use HP inkjets and laserjets and an Epson 600.
The HP inkjets to a fair to middling job of either color or black and white (true for all the Deskjet 6XX models I've used - 600, 660). Their fast and cheap and not bad if you're not at all picky. The pictures are grainy.
I have an HP Laserjet 5P I use for black and white. At the present time we usually give a polaroid print to the person requesting work and laserjet copies (four to a page which makes the laserjet "prints" just about the same size as the polaroids). The laserjet pictures are not by any stretch of the imagination "photoquality" but they are very good and give more than enough detail for most purposes.
The Epson 600 is used the same way as the laserjet for color photos. Again, these are not "photoquality", but they are amazingly good even at 720X720 dpi. However, even at 720 dpi, it is excruciatingly slow. I think Epson got it's fraction turned around when they figured out the page rate. I think they quote something like 2 pages a minute. 2 minutes per page would be far closer to the truth.
All of this is on the laserjet paper they use around here for normal office work for the laserjet or good (not the really good shiny stuff) inkjet paper for the Epson. If I started using the really good stuff I suspect the results would be noticably better but so would the cost per page which was the point in going to these printers in the first place.
Ive never figured out the actual cost per page for any of these. I'm pretty sure it's far below the $2/print I pay for B&W polaroid or $.75/print for video printers.
============================================================ Bede Willenbring Phone: (651)236-5470 H.B. Fuller Company FAX: (651)236-5430 Research Chemist E-mail: Bede.Willenbring-at-HBFuller.com P.O. Box 64683 St. Paul, MN 55164-0683 ___________________________________________
B&W printer are actually harder to find than good color ones - the Lexmark 5700, Epson Photo ot Cannon 7004 all are great color printers - (6 or 7 colors and } 1000 dpi).
Bill Miller ___________________________________________
We have both. HP LaserJet's which we use for viewing B&W images. The 600 dpi output is more than good enough for most work and we seldom go to film anymore. You won't have any trouble getting someone who has one to print a BMP or other photo out to show you how good they look. I also have an Epson 1440x720 dpi color printer for near photo quality in color, and I seldom use it for that, although I just bought a digital camera. I also have a 300 dpi HP 1600color inkjet which I use the heck out of because its fast and good enough for initial work like cropping and framing and positioning. Film is fast becoming sidelined in all the branches of our company, and I see the same thing wherever I go. I am getting a 1200 dpi LaserJet with my next machine so my B&Ws will look even better. The metallurgists are jealous because they are stuck with an expensive, slow photo printing system which uses proprietary file formats. They will be upgraded someday.
I don't know anything about Alps. HP is so easy to set up (network too) and has the fastest drivers, Epson had to outdo them big in dpi to get my attention.
Tim Chavez Boeing Chem Characterization Lab ___________________________________________
We have a HP 890C Inkjet and you can't beat it. It is has the Kodak PhotoRes processing and really does wonderful output. It is as good as a sublimation dye printer (we have two) when the special Kodak Photo Deluxe paper is used. It is very good on regular paper, better still on the HP premium paper. The printer is about $360. You can get a 722C for about $260 which uses the same cartridges and is a little slower. There are web sites that you can find the cost of the media and ink cartridges. Several people here who have Epson inkjet have been using ours to print critical stuff and slides because the quality is so much better. Don't have experience on the Alps.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
___________________________________________
We recently purchased a Kodak SP700 dye sub printer. The thing takes a couple minutes per print, and prints only on their specialized paper, but the cost is low per print ( {70 cents) and does a very good gray scale rendition.
We use an Epson 600, a Lexmark Optra Rt+ and a HP Laserjet 4M for general purpose printing. it is enough for most people. If they want a real good print we will print on the Kodak or send the image back to our SEM photo CRT (special hardware required).
Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
We are using the Epson Photo and are very pleased with the results. Cost per print is dependent on the type of paper you use. We do proofs on laser paper and use injet matte or glossy for better quality prints. Publication prints are usually done on a dye sub printer, although the glossy inkjet prints look awfully good. I could send samples if you like.
Greg
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax:352-846-0251 University of Florida Gainesville, FL 32611
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All } } I have a series of tiff secondary electron images of serial sections } through a polished mineral grain. Could someone please advise me as to the } best software available for doing 3-D reconstructions from these tiff files } on my MAC (266mHz G3). Each is around 1 meg and they are about 60 } sequential images. } } Thank you } } Brendan } } Brendan J. Griffin } Centre for Microscopy and Microanalysis } The University of Western Australia } Nedlands, WA, AUSTRALIA 6907 } ph 61-8-9380-2739 fax 61-8-9380-1087 } } bjg-at-cyllene.uwa.edu.au } } }
I used a freeware available on the net, nIH Image. The results??, well you can see them for your selv...
Gary...
On Fri, 23 Oct 1998, Brendan Griffin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All } } I have a series of tiff secondary electron images of serial sections } through a polished mineral grain. Could someone please advise me as to the } best software available for doing 3-D reconstructions from these tiff files } on my MAC (266mHz G3). Each is around 1 meg and they are about 60 } sequential images. } } Thank you } } Brendan } } Brendan J. Griffin } Centre for Microscopy and Microanalysis } The University of Western Australia } Nedlands, WA, AUSTRALIA 6907 } ph 61-8-9380-2739 fax 61-8-9380-1087 } } bjg-at-cyllene.uwa.edu.au } } }
I agree Woody. I think Leco makes a cold mounting material (LecoSet) that outgases till the cows come home. And you can smell it from the next room as well. Hysol makes an epoxy resin that doesn't seem to outgas at all. Other than the continuity problems encountered...it does work well. How about these carbon or copper filled thermosetting Bakelite materials? We use them...however they do create problems when we electrolytically polish/etch metallic samples in those mounts. Our voltage settings for the electropolisher sense the continuity of the mounting material so that caused us to change our parameters.... .....an environmental SEM is in our future here.
Harry A. Ekstrom AlliedSignal Inc.
---------- } From: "Woody.N.White-at-mcdermott.com"-at-sparc5.microscopy.com To: Microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
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Reply to: RE: TEM:microwave immunocytochemistry B. Laube wrote: } } Dear all, does anybody have experience with immunocytochemistry on Ni-, = Au- grids with a = } temperature probe controlled laboratory microwave oven? Is it also time = consuming and one = } could achieve better or comparable staining results protocols? I think, = one problem could be the = } irradiation of the metallic grids, leading to overheating spots on the = grid bars and this may = } precipitate the antibodies which would cause useless results. Is this = negligible? =
Reply: It is tempting to think that the improved penetration of fixatives caused = by microwave irradiation could help with antibody penetration into = sections. Indeed there are many published reports of "antigen retrieval" = using microwaves for light microscopy.
We just completed a study of the effects of microwaves on antibody = labeling for EM (1998 Microscopy Research and Technique 42:24-32) and = concluded that although there is sometimes an increase in labeling density,= there are so many variables (antibody, antigen, tissue, fixative etc) = that the method cannot yet be applied routinely. We were able to = irradiate Ni or Cu grids in the microwave oven without problem, so go = ahead and try it. We did find that if protein A-gold is irradiated, the = signal is reduced considerably. Similarly with some blocking agents.
When comparing labeling times, to determine if they can be reduced in = microwave ovens, make sure you perform the most obvious control of taking = away the microwaves. In our study, we did find that the total labeling = time could be cut to 30 min or less for some antibodies. However, when = the microwave oven was taken away, we obtained almost identical results!
For some systems, we did get enhanced labeling which could easily be = explained as improved penetration of antibodies. However, as it is almost = universally accepted that there is almost no penetration of antibodies = into intact sections, a manuscript describing improved labeling over = Lowicryl sections did bother us (I can find the reference if anyone is = interested). One possible explanation for this observation may come from = one of our preliminary experiments. For one of our antibodies we found = that labeling efficiency improved dramatically after it had been exposed = to microwaves. Neither the sections or other reagents had been exposed.
In conclusion, microwaves do offer advantages to biological microscopists (= especially for fixation and embedding, as well as for more specialized = extraction methods) and may yet offer substantial benefits in reducing = antibody labeling times or by increasing labeling efficiency. However, = more experimentation is required with extremely rigorous and critical = evaluation. Putting an experiment into the microwave oven and then = claiming improved results is not enough any more. =
I wait for someone to tell me that Cu grids cannot be used for = immunocytochemistry! =
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org
by gate.mcdermott.com (8.8.7/8.8.7) with ESMTP id SAA04067 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 23 Oct 1998 18:02:36 -0500 (CDT) Received: (from x400-at-localhost) by mta.mcdermott.com (8.7.1/8.7.1) id SAA00805 for Microscopy-at-MSA.Microscopy.Com; Fri, 23 Oct 1998 18:04:14 -0500 (CDT) X-Authentication-Warning: mta.mcdermott.com: x400 set sender to Woody.N.White%650-at-wtgw.mcdermott.com using -f Received: by MCI; Fri, 23 Oct 1998 15:10:00 -0500
I feel that accurate quant of carbon occuring in less than 1 percent concentrations (most steels) will be nearly impossible, even with the latest and greatest equipment. For example, if the steel is in a mount and you don't have a dry pumped vacuum system, the carbon deposits on the surface are likely yield a larger carbon peak after a few minutes than would the carbon from the alloy.
Anyone out there more optimistic??
Woody White McDermott Technology, Inc.
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Is anyone here using EDX to measure the amount and depth of decarburization of steel? I'd like to analyze the depth of the decarburization of steel that has been through a furnace. I'd like to take a cross section of the slug and beginning at the outer edge, perform a semiquant analysis with my EDX at succeeding depths from the outer edge into the core, measuring for carbon content. Is anyone here using an SEM for this type of work? I'd like to know how you go about it and what the success and confidence level is.
Dear Mark, If you look at the levels of carbon in these steel samples, they are too low for meaningful analysis. I have tried many times and never been able to determine the difference in carbon content over the entire range possible in steel. I can just detect the carbon in coarse pearlite (Fe3C) on my WDX, with difficulty. You are welcome to try, good luck. The sample cannot be mounted or the carbon in the mounting material will swamp the signal. You wrote: } } Is anyone here using EDX to measure the amount and depth of } decarburization of steel? I'd like to analyze the depth of the } decarburization of steel that has been through a furnace. } I'd like to take a cross section of the slug and beginning at the outer } edge, perform a semiquant analysis with my EDX at succeeding depths } from the outer edge into the core, measuring for carbon content. } Is anyone here using an SEM for this type of work? I'd like to know } how you go about it and what the success and confidence level is. } } Thanks } } Mark Darus } BFGoodrich, Landing Gear Division } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
I have had some success analyzing carbon by using WDS on an electron microprobe.
However, this required: a) LN2 trap above the diffusion pump b) Air jet on sample to burn off contamination as beam sits on a location c) LN2 cold trap above the sample d) Calibration curve made from measurements of count rates from a set of four steel samples with C wt.% ranging between 0 and 1.29% e) Careful cleaning of samples f) Beam current regulation g) Careful selection of background locations h) Would work with either a lead stearate pseudocrystal or multilayer synthetic reflector, though the latter will give better count rates. i) High beam current (100nA), low accelerating voltage (10kV), long counting times (at least 60 seconds) and usually a rastered beam (at least 6 x 6 um, but no bigger than about 30 x 30 um) to integrate microinclusions.
I would not be optimistic about trying this with EDS or on a standard SEM.
On Fri, 23 Oct 1998 Woody.N.White-at-mcdermott.com-at-sparc5.microscopy.com wrote: : : I feel that accurate quant of carbon occuring in less than 1 percent : concentrations (most steels) will be nearly impossible, even with the latest : and : greatest equipment. For example, if the steel is in a mount and you don't : have a : dry pumped vacuum system, the carbon deposits on the surface are likely : yield a : larger carbon peak after a few minutes than would the carbon from the alloy. : : : Anyone out there more optimistic?? : : Woody White : McDermott Technology, Inc. : : : Is anyone here using EDX to measure the amount and depth of : decarburization of steel? I'd like to analyze the depth of the : decarburization of steel that has been through a furnace. : I'd like to take a cross section of the slug and beginning at the outer : edge, perform a semiquant analysis with my EDX at succeeding depths : from the outer edge into the core, measuring for carbon content. : Is anyone here using an SEM for this type of work? I'd like to know : how you go about it and what the success and confidence level is. : : Thanks : : Mark Darus : BFGoodrich, Landing Gear Division :
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA -------------------------------- Voice: (734) 936-1550 FAX: (734) 763-4690 E-mail: chender-at-umich.edu --------------------------------
This procedure is very difficult to accomplish accurately even with an Electron Microprobe. Getting a carbon standard with certified, small amounts of carbon is necessary. Maybe with these new EDX detectors today and the latest software, digital pulse processors, etc....it can be done with an SEM. How accurate are they tho? I won't venture to say. Any other opinions out there?
Harry Ekstrom
---------- } From: Mark Darus To: microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
On Fri, 23 Oct 1998, Carl Henderson wrote: : I have had some success analyzing carbon by using WDS on an electron : microprobe. : : However, this required:
: g) Careful selection of background locations
Right after I sent this message, I remembered that I did take background counts. Since I used a calibration curve, this was not necessary (one of the points on the curve is from a 0.0 wt% C standard).
Carl
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA -------------------------------- Voice: (734) 936-1550 FAX: (734) 763-4690 E-mail: chender-at-umich.edu --------------------------------
This might be irrelevent for this listserver. But this fruad seems a nationwide phenomenon and very close to everybody.
} -----Original Message----- } From: Cindy Euton } Sent: Tuesday, October 20, 1998 10:05 AM } To: EVERYONE } Subject: FW: Phone Fraud } [Cindy Euton] This was forwarded to me by a friend, so watch out for } these kinds of scams } Hi, All, } I received a telephone call today from an individual identifying himself } as an AT&T Service Technician who was conducting a test on our telephone } lines. He stated that to complete the test I should touch nine (9), zero } (0), the pound sign (#) and then hang up. Luckily, I was suspicious and } refused. Upon contacting the telephone company, I was informed that by } pushing 90#, you give the requesting individual full access to your } telephone line, which allows them to place long distance telephone calls } billed to your phone number. I was further informed that this scam has } been originating from many of the local jails/prisons. I have also } verified this information with UCB Telecomm, Pacific Bell, MCI, Bell } Atlantic, GTE and NYNEX. Please beware. DO NOT press 90# for ANYONE. } The GTE Security Department requested that I share this information with } EVERYONE I KNOW. PLEASE pass this on to everyone YOU know. If you have } mailing lists and/or newsletters from organizations you are connected } with, I encourage you to pass on this information to them, too. Thought } you might like to know.... } Also, I called Southwestern Bell (713) 638-7200 and they verified that } this is true. } } } } } }
Dear Microscopists: Gary Login has cited some very good papers on using the microwave. The microwave does provide faster fixation, staining of tissues, and also allows antigen retrieval in parafin embedded sections. Many of the techniques he refers to put sections in the microwave which are labeled conventionally after irradiation. It seems that microwaving the samples allows access in parafin embedded sections. It is also accepted that the fixation from microwaves is as good as conventional fixation methods which can save time. However, in the same respect, Paul Webster's response refers to labeling of antibodies inside the microwave which does not provide reproducible results from one system to another. Because the antibodies are microwave irradiated, they could be affected by the irradiation and may not provide accurate labeling densities. Linda Chicoine Cognetix/Viatech, Inc. Ivoryton, CT
A major research center has inquired about us participating in a project requiring a SEM with a computer-driven stage. Our SEMs have only the standard manual stages. If your SEM has a computer-driven stage and you accept outside work, please contact me and I will put you in direct contact with the researcher.
J. Roy Nelson Material Testing Laboratory Pennington, NJ (609) 730-0575 jrnelson-at-nj1.aae.com
As a newcomer to this listserver, I appreciate the quick responses to my dilemma in locating light bulbs for my Durst enlarger. I have not yet made all the contacts recommended but am impressed with your cooperation. Thank you!
Mary North Loma Linda UMC Loma Linda, CA northstar44-at-juno.com
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My pathologist seems to remember an acid maltase stain performed on a frozen muscle section (instead of biochemical analysis on a larger portion). If this technic is available, please forward the information to me at this listserver or my e-mail address.
Mary North Loma Linda University Medical Center Loma Linda, CA northstar44-at-juno.com
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
I'd like to ditto Wolf's comments, unless you have some 3D background, start out with something free (NIH-Image), then look at what is commercially available. I really recommend checking out ObjectImage too. Importantly, and often overlooked, check out the NIH-Image discussion group and archive, your questions have probably been previously addressed there but do subscribe to the discussion. I've found the imaging tips and techniques priceless.
I purchased VoxBlast and IDL and really like both, but your success with them will depend on what your goals are, both have demos available. But remember, IDL has a steeper learning curve than the other programs so it helps if you've programmed before, though there are very good resources for learning (classes, discussion newsgroup, David Fanning's book at http://www.dfanning.com/). IDL can give you more options, but you will probably have to program to make it happen, and like VoxBlast, it runs on many platforms. I think many would agree there is no one 3-D program which does it all, therefore familiarity with several will benefit you most. Also, be sure to check out John Russ's book (CRC Press) and Image Processing Toolkit software. John has been around imaging a long time and knows it well (unpaid endorsement).
I've also found organizations like SPIE (http://www.spie.org/) useful resources concerning applications to medical/biomedical imaging (annual meeting of medical imaging is neat-o), including volume visualization and spatial analysis - beyond 3D reconstruction but it seems to me much more informative for the work of reconstructing large datasets. I've found many connections between the techniques of volume visualization in radiological applications can be applied to microscopically generated 3-D volumes.
good luck,
} } Hi Chris, } } I recommend to start working slowly. Why don't you get yourself the FREE } software NIH IMAGE. It is not only free, but it is the standard. } } http://rsb.info.nih.gov } } Look out, also, for the also free modification called OBJECT IMAGE, } which is even better. Also on one of those servers. Just don't forget to } ASSIGN ENOUGH MEMORY to those programs before you start digging around in } your huge files. } } They can be both used to reslice the 3D block (then called stack). Object } Image also can interactively BROWSE through your block in three planes at } once } in real time. This gives you a pretty cool idea of what things there might } be to visualize. } } Don't spend too much money before you know what you need, get organized } with the 3D possibilites, and more important, the diagnostic relevance or } irrelevance of some features, and look out for the capabilities of IDL, the } BEST interactive graphics tool around (Interactive Data Language, } Research Systems, www.rsinc.com). } } This is independent advice. - Cheers Wolf } }
Brian C. Tryon Medical Student Allegheny University of the Health Sciences School of Medicine, Ann Preston Hall, Box 551 3300 Henry Avenue Philadelphia, PA 19129 USA
----------------------------------------- "Quantifying is a committing task." - Cruz-Orive, 1994.
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." - Richard Feynman --------------------------------------------------------------
Dear all I have been testing quite a few resins for mounting and polishing metallurgical based samples for lightmicroscopy as well as SEM observation and EDS analysis. Important criteria is edge rotention and beamstability. The best two resins I found was Bakelite (black powder hot press resin) and Araldite for cold mounting. Araldite (the cheap version) can be bought from moust shops selling fiberglass kits. Some of the other resins I hate for either being smelly, beam instable, or bad edge rotention or just to costly for student use. } } I have also had good luck with epofix as well as hot pressed } (thermoset) resins. As stated by Bigelow, it is very important to } mix and cure properly to avoid outgasing. } } Certainly not all resins are created equal. I forget the name } (mental block?), but a few years ago we experimented with a bubble } gum pink mount. It took a month at 120F in a vacuum oven before I } could reach 1x10-3 torr! Heard it was discontinued - Thank } goodness. Could smell the mount... If you can smell it, forget } it:) } } Woody White } McDermott Technology, Inc } ----------------------------------------------------------------- } } My question: } What metallographic mounting materials are safe for an SEM? Some of our } users would like us to forbid usage of any mounting compound other than } those } that require both pressure and heat such as bakelite and its near relatives. } } They feel that the resolution of the machine will gradually degrade if we } allow the usage of cold mounting materials. We have currently allowing the } usage of some of the more stable (epoxy from leco and buehler for example) } cold mounts. We don't see any change in vacuum when the beam strikes the } sample mount but are we slowly degrading the image resolution by gunking up } the column? To date the resolution still is fine but what about the future?! } } } Thanks, } } Robin Griffin } } Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
Dear all I have been testing quite a few resins for mounting and polishing metallurgical based samples for lightmicroscopy as well as SEM observation and EDS analysis. Important criteria is edge rotention and beamstability. The best two resins I found was Bakelite (black powder hot press resin) and Araldite for cold mounting. Araldite (the cheap version) can be bought from moust shops selling fiberglass kits. Some of the other resins I hate for either being smelly, beam instable, or bad edge rotention or just to costly for student use. } } I have also had good luck with epofix as well as hot pressed } (thermoset) resins. As stated by Bigelow, it is very important to } mix and cure properly to avoid outgasing. } } Certainly not all resins are created equal. I forget the name } (mental block?), but a few years ago we experimented with a bubble } gum pink mount. It took a month at 120F in a vacuum oven before I } could reach 1x10-3 torr! Heard it was discontinued - Thank } goodness. Could smell the mount... If you can smell it, forget } it:) } } Woody White } McDermott Technology, Inc } ----------------------------------------------------------------- } } My question: } What metallographic mounting materials are safe for an SEM? Some of our } users would like us to forbid usage of any mounting compound other than } those } that require both pressure and heat such as bakelite and its near relatives. } } They feel that the resolution of the machine will gradually degrade if we } allow the usage of cold mounting materials. We have currently allowing the } usage of some of the more stable (epoxy from leco and buehler for example) } cold mounts. We don't see any change in vacuum when the beam strikes the } sample mount but are we slowly degrading the image resolution by gunking up } the column? To date the resolution still is fine but what about the future?! } } } Thanks, } } Robin Griffin } } Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
Try something pretty standard before going to the exotics like a slow fix with gradually increasing concentrations of Glut. A possibility: try 2% Glutaraldehyde in 0.05M Phosphate with 4% Sucrose for 2 h Postfix in 1% OsO4 for 1h. Embed as usual. A reference is: Kaidoh et al - 1993, J. Euk. Microbiol. 40(3) 269-271.
Jan C
} } } If anyone out there has done work with Malaria, and knows of the best } TEM fixative for this little RBC parasite, please contact me at: } lamiller-at-uiuc.edu } } } Thanks! } } Lou Ann } }
Prof Jan Coetzee Head: Lab for Microscopy and Microanalysis Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-362-5150 Pretoria 0002, South Africa http://www.up.ac.za/science/electron/emunit1.htm
Yep, I can see where the increased area, even if at lower average conductivity would present problems in achieving the proper current density.
For (bulk) conductive mounting material, Struers copper loaded was my favorite. Obviously, edges presented problems due to the "islands" of polymer between the Cu. I was able clean the mounts with both water/detergent and alcohol with little problem. The carbon loaded mount material we had was not so "nice" about that. The met lab would prep a sample for me, look at it optically, and think it was clean. Never made a study of it, but apparently something would slightly dissolve in alcohol, leaving an optically transparent film that was almost impossible to remove. Samples looked "cloudy" in the sem. I specify that carbon material not be used now.
I am told that Struers discontinued the Cu material. Do you know a source?
I have, with limited success, made conductive cold-set, by adding a heavy concentration of zinc dust to the polymer mix. Beware! Metal dusts can be reactive/explosive. Be sure, if this is attempted, to predetermine any hazard.
What I wish for is a thermosetting intrinsically conductive polymer. All I have found so far is ground/powdered/cured material which cannot be hot pressed. I believe an Allied Signal division made some of this....
Regards,
Woody White McDermott technology, Inc.
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I agree Woody. I think Leco makes a cold mounting material (LecoSet) that outgases till the cows come home. And you can smell it from the next room as well. Hysol makes an epoxy resin that doesn't seem to outgas at all. Other than the continuity problems encountered...it does work well. How about these carbon or copper filled thermosetting Bakelite materials? We use them...however they do create problems when we electrolytically polish/etch metallic samples in those mounts. Our voltage settings for the electropolisher sense the continuity of the mounting material so that caused us to change our parameters.... ....an environmental SEM is in our future here.
Harry A. Ekstrom AlliedSignal Inc.
---------- } From: "Woody.N.White-at-mcdermott.com"-at-sparc5.microscopy.com To: Microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
I don't know if you answer stuff like this but I'm kind of stuck at the moment. I'm looking for information on the investigators involved in the development of the TEM but am unable to find any information anywhere on the web. If you know of anywhere this information may be found I would appreciate it a lot.
If you need only surface renderings and not internal volume or reslicing ability, I can recommend SURFdriver. This was developed by two anatomists specifically to do 3D tissue reconstructions from histological serial sections, but also works with other 3D data. Easy to use and does quick renderings. A free demo version is available at http://surfdriver.ml.org.
I have no connection with SURFdriver other than as a satisfied user.
Gary Radice 804-289-8107 Department of Biology 804-289-8233 (FAX) University of Richmond gradice-at-richmond.edu Richmond VA 23173
There IS an adhesive that is: smooth-surfaced, high tack, low Z, durable, easy to use, dependable, nontoxic and inexpensive. It is also pretty stable under the beam and thin layers do not outgas noticeably in the vacuum. It is, however, not conductive (after all - is there such a thing as a perfect whatever?). We use this stuff regularly to stick down pollen, fly-ash, small insects, blood cells, ceramic powders, pigment particles etc. for imaging and for EDS analysis.
it is called 'Artists Gold Size' and is a thick tacky syrup of partially polymerised liseed oil. On exposure to air it polymerizes and forms an insoluble polymer (this was used in the manufacture of old-style floor linoleum, hence the name linoleum - from 'linseed oleum').
Buy this at any art supply shop, spread a very small drop of the syrup over the surface of a stub, wait until it is just tacky enough not to wick up the sample, drop your volcanic ash particles onto the surface and leave for 30 min. at room temp (or a few min at 50C) to polymerize, coat with carbon and analyse.
I've never been able to find out who used this first, but have been using it for many years after coming across a casual reference to its use as a SEM mountant.
A $5 bottle should last you years.
Pity it isn't conductive though.
Jan C
} } I'd really like to have a durable, very smooth, low Z surface, that is } insensitive to beam current and is easy to apply and work with in the SEM. } Oh, it should also be economical too. } } Any ideas? TIA } } Owen
Prof Jan Coetzee Head: Lab for Microscopy and Microanalysis Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-362-5150 Pretoria 0002, South Africa http://www.up.ac.za/science/electron/emunit1.htm
WARNING! WARNING! DANGER! In case you can not tell from the line below which states that "...we have available...", I have a vested interest in this. I am trying to offer you something for sale (GASP!) which you may be interested in. If you are horrified by advertising, please read no further! (unless you are trying to get a good scare in time for Halloween).
I didn't see the originial inquiry, but let me state that we have available a dye sublimation printer for $2300. Print quality is on par with the $5-9K units, and far better than you will ever see from an inkjet printer. You will get photograph quality prints from the printer. If you are interested in futher details, plese email me or call me using the information below.
Sincerely, J. Haley
****************************** Jim Haley Applications Engineer I-CUBE 2411 Crofton Lane, Suite 14A Crofton, MD 21114 voice: (301) 858-0505 fax: (301) 858-0615 web site: http://www.i-cubeinc.com e-mail: haley-at-i-cubeinc.com ******************************
drose-at-wlgore.com-at-Sparc5.Microscopy.Com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello List, } } Thanks to all who responded to my posting. All the information has been } helpful. Responses are summarized below. } } David Rose } W.L. Gore and Associates } } =========================================== } } I've seen posts which responded to the original "HP vs Epson vs Alps" } which imply Alps is an ink jet. The original poster may have been } referring to the Alps 1300 which is a dual mode 600dpi ink jet and } dye-sub printer for ~$500. I was personally impressed with the example } Alps sent me, but I also hear it may be the slowest dye-sub on the } market ... and we all know printer performance has more to do with how } well the printer interfaces with the OS. I know nothing beyond this ... } but certainly am curious if someone has practical experience. } } cheerios, shAf } } {} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} } Michael Shaffer, R.A. - ICQ 210524 } Geological Science's Electron Probe Facility - University of Oregon } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/ } } ___________________________________________ } } Only real photo quality is dye sublimation (kodak, codonics, tektronics, } etc) } which are expensive (maybe $5K and up). Inkjets like you name, with } special, } expensive glossy paper and 1200 dpi are almost as good for a few hundred $. } I } like the Epson Stylus Photo (EX) myself. All will be color (but make good } B&W) } unless you get a laser printer. Dave Pevear, Houston } } ___________________________________________________ } } I can't tell you anything about Alps, but I do use HP inkjets and laserjets } and an Epson 600. } } The HP inkjets to a fair to middling job of either color or black and white } (true for all the Deskjet 6XX models I've used - 600, 660). Their fast and } cheap and not bad if you're not at all picky. The pictures are grainy. } } I have an HP Laserjet 5P I use for black and white. At the present time } we usually give a polaroid print to the person requesting work and laserjet } copies (four to a page which makes the laserjet "prints" just about the } same size as the polaroids). The laserjet pictures are not by any stretch } of the imagination "photoquality" but they are very good and give more than } enough detail for most purposes. } } The Epson 600 is used the same way as the laserjet for color photos. } Again, these are not "photoquality", but they are amazingly good even at } 720X720 dpi. However, even at 720 dpi, it is excruciatingly slow. I think } Epson got it's fraction turned around when they figured out the page rate. } I think they quote something like 2 pages a minute. 2 minutes per page } would be far closer to the truth. } } All of this is on the laserjet paper they use around here for normal office } work for the laserjet or good (not the really good shiny stuff) inkjet } paper for the Epson. If I started using the really good stuff I suspect } the results would be noticably better but so would the cost per page which } was the point in going to these printers in the first place. } } Ive never figured out the actual cost per page for any of these. I'm } pretty sure it's far below the $2/print I pay for B&W polaroid or } $.75/print for video printers. } } ============================================================ } Bede Willenbring Phone: (651)236-5470 } H.B. Fuller Company FAX: (651)236-5430 } Research Chemist E-mail: Bede.Willenbring-at-HBFuller.com } P.O. Box 64683 } St. Paul, MN 55164-0683 } ___________________________________________ } } B&W printer are actually harder to find than good color ones - the Lexmark } 5700, Epson Photo ot Cannon 7004 all are great color printers - (6 or 7 } colors and } 1000 dpi). } } Bill Miller } ___________________________________________ } } We have both. } HP LaserJet's which we use for viewing B&W images. The 600 dpi output is } more than good enough for most work and we seldom go to film anymore. } You won't have any trouble getting someone who has one to print a BMP or } other photo out to show you how good they look. } I also have an Epson 1440x720 dpi color printer for near photo quality } in color, and I seldom use it for that, although I just bought a digital } camera. I also have a 300 dpi HP 1600color inkjet which I use the heck } out of because its fast and good enough for initial work like cropping } and framing and positioning. Film is fast becoming sidelined in all the } branches of our company, and I see the same thing wherever I go. I am } getting a 1200 dpi LaserJet with my next machine so my B&Ws will look } even better. The metallurgists are jealous because they are stuck with } an expensive, slow photo printing system which uses proprietary file } formats. They will be upgraded someday. } } I don't know anything about Alps. HP is so easy to set up (network too) } and has the fastest drivers, Epson had to outdo them big in dpi to get } my attention. } } Tim Chavez } Boeing Chem Characterization Lab } ___________________________________________ } } We have a HP 890C Inkjet and you can't beat it. It is has the Kodak } PhotoRes processing and really does wonderful output. It is as good as a } sublimation dye printer (we have two) when the special Kodak Photo Deluxe } paper is used. It is very good on regular paper, better still on the HP } premium paper. The printer is about $360. You can get a 722C for about } $260 which uses the same cartridges and is a little slower. There are web } sites that you can find the cost of the media and ink cartridges. Several } people here who have Epson inkjet have been using ours to print critical } stuff and slides because the quality is so much better. Don't have } experience on the Alps. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Guys Run Rd. (packages) } P.O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } ___________________________________________ } } We recently purchased a Kodak SP700 dye sub printer. The thing takes a } couple minutes per print, and prints only on their specialized paper, but } the cost is low per print ( {70 cents) and does a very good gray scale } rendition. } } We use an Epson 600, a Lexmark Optra Rt+ and a HP Laserjet 4M for general } purpose printing. it is enough for most people. If they want a real good } print we will print on the Kodak or send the image back to our SEM photo } CRT (special hardware required). } } Warren E. Straszheim } 23 Town Engineering } Iowa State University } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } http://www.marl.iastate.edu } } ___________________________________________ } } We are using the Epson Photo and are very pleased with the results. Cost } per print is dependent on the type of paper you use. We do proofs on laser } paper and use injet matte or glossy for better quality prints. Publication } prints are usually done on a dye sub printer, although the glossy inkjet } prints look awfully good. I could send samples if you like. } } Greg } } Gregory W. Erdos, Ph.D. Ph. 352-392-1295 } Assistant Director, Biotechnology Program } PO Box 110580 Fax:352-846-0251 } University of Florida } Gainesville, FL 32611
WARNING! WARNING! DANGER! In case you can not tell from the line below which states that "...we have available...", I have a vested interest in this. I am trying to offer you something for sale (GASP!) which you may be interested in. If you are horrified by advertising, please read no further! (unless you are trying to get a good scare in time for Halloween).
I didn't see the originial inquiry, but let me state that we have available a dye sublimation printer for $2300. Print quality is on par with the $5-9K units, and far better than you will ever see from an inkjet printer. You will get photograph quality prints from the printer. If you are interested in futher details, plese email me or call me using the information below.
Sincerely, J. Haley
****************************** Jim Haley Applications Engineer I-CUBE 2411 Crofton Lane, Suite 14A Crofton, MD 21114 voice: (301) 858-0505 fax: (301) 858-0615 web site: http://www.i-cubeinc.com e-mail: haley-at-i-cubeinc.com ******************************
} drose-at-wlgore.com-at-Sparc5.Microscopy.Com wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hello List, } } } } Thanks to all who responded to my posting. All the information has been } } helpful. Responses are summarized below. } } } } David Rose } } W.L. Gore and Associates } } } } =========================================== } } } } I've seen posts which responded to the original "HP vs Epson vs Alps" } } which imply Alps is an ink jet. The original poster may have been } } referring to the Alps 1300 which is a dual mode 600dpi ink jet and } } dye-sub printer for ~$500. I was personally impressed with the example } } Alps sent me, but I also hear it may be the slowest dye-sub on the } } market ... and we all know printer performance has more to do with how } } well the printer interfaces with the OS. I know nothing beyond this ... } } but certainly am curious if someone has practical experience. } } } } cheerios, shAf } } } } {} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} } } Michael Shaffer, R.A. - ICQ 210524 } } Geological Science's Electron Probe Facility - University of Oregon } } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/ } } } } ___________________________________________ } } } } Only real photo quality is dye sublimation (kodak, codonics, tektronics, } } etc) } } which are expensive (maybe $5K and up). Inkjets like you name, with } } special, } } expensive glossy paper and 1200 dpi are almost as good for a few hundred $. } } I } } like the Epson Stylus Photo (EX) myself. All will be color (but make good } } B&W) } } unless you get a laser printer. Dave Pevear, Houston } } } } ___________________________________________________ } } } } I can't tell you anything about Alps, but I do use HP inkjets and laserjets } } and an Epson 600. } } } } The HP inkjets to a fair to middling job of either color or black and white } } (true for all the Deskjet 6XX models I've used - 600, 660). Their fast and } } cheap and not bad if you're not at all picky. The pictures are grainy. } } } } I have an HP Laserjet 5P I use for black and white. At the present time } } we usually give a polaroid print to the person requesting work and laserjet } } copies (four to a page which makes the laserjet "prints" just about the } } same size as the polaroids). The laserjet pictures are not by any stretch } } of the imagination "photoquality" but they are very good and give more than } } enough detail for most purposes. } } } } The Epson 600 is used the same way as the laserjet for color photos. } } Again, these are not "photoquality", but they are amazingly good even at } } 720X720 dpi. However, even at 720 dpi, it is excruciatingly slow. I think } } Epson got it's fraction turned around when they figured out the page rate. } } I think they quote something like 2 pages a minute. 2 minutes per page } } would be far closer to the truth. } } } } All of this is on the laserjet paper they use around here for normal office } } work for the laserjet or good (not the really good shiny stuff) inkjet } } paper for the Epson. If I started using the really good stuff I suspect } } the results would be noticably better but so would the cost per page which } } was the point in going to these printers in the first place. } } } } Ive never figured out the actual cost per page for any of these. I'm } } pretty sure it's far below the $2/print I pay for B&W polaroid or } } $.75/print for video printers. } } } } ============================================================ } } Bede Willenbring Phone: (651)236-5470 } } H.B. Fuller Company FAX: (651)236-5430 } } Research Chemist E-mail: Bede.Willenbring-at-HBFuller.com } } P.O. Box 64683 } } St. Paul, MN 55164-0683 } } ___________________________________________ } } } } B&W printer are actually harder to find than good color ones - the Lexmark } } 5700, Epson Photo ot Cannon 7004 all are great color printers - (6 or 7 } } colors and } 1000 dpi). } } } } Bill Miller } } ___________________________________________ } } } } We have both. } } HP LaserJet's which we use for viewing B&W images. The 600 dpi output is } } more than good enough for most work and we seldom go to film anymore. } } You won't have any trouble getting someone who has one to print a BMP or } } other photo out to show you how good they look. } } I also have an Epson 1440x720 dpi color printer for near photo quality } } in color, and I seldom use it for that, although I just bought a digital } } camera. I also have a 300 dpi HP 1600color inkjet which I use the heck } } out of because its fast and good enough for initial work like cropping } } and framing and positioning. Film is fast becoming sidelined in all the } } branches of our company, and I see the same thing wherever I go. I am } } getting a 1200 dpi LaserJet with my next machine so my B&Ws will look } } even better. The metallurgists are jealous because they are stuck with } } an expensive, slow photo printing system which uses proprietary file } } formats. They will be upgraded someday. } } } } I don't know anything about Alps. HP is so easy to set up (network too) } } and has the fastest drivers, Epson had to outdo them big in dpi to get } } my attention. } } } } Tim Chavez } } Boeing Chem Characterization Lab } } ___________________________________________ } } } } We have a HP 890C Inkjet and you can't beat it. It is has the Kodak } } PhotoRes processing and really does wonderful output. It is as good as a } } sublimation dye printer (we have two) when the special Kodak Photo Deluxe } } paper is used. It is very good on regular paper, better still on the HP } } premium paper. The printer is about $360. You can get a 722C for about } } $260 which uses the same cartridges and is a little slower. There are web } } sites that you can find the cost of the media and ink cartridges. Several } } people here who have Epson inkjet have been using ours to print critical } } stuff and slides because the quality is so much better. Don't have } } experience on the Alps. } } } } -Scott } } } } Scott D. Walck, Ph.D. } } PPG Industries, Inc. } } Guys Run Rd. (packages) } } P.O. Box 11472 (letters) } } Pittsburgh, PA 15238-0472 } } } } Walck-at-PPG.com } } } } (412) 820-8651 (office) } } (412) 820-8161 (fax) } } } } ___________________________________________ } } } } We recently purchased a Kodak SP700 dye sub printer. The thing takes a } } couple minutes per print, and prints only on their specialized paper, but } } the cost is low per print ( {70 cents) and does a very good gray scale } } rendition. } } } } We use an Epson 600, a Lexmark Optra Rt+ and a HP Laserjet 4M for general } } purpose printing. it is enough for most people. If they want a real good } } print we will print on the Kodak or send the image back to our SEM photo } } CRT (special hardware required). } } } } Warren E. Straszheim } } 23 Town Engineering } } Iowa State University } } Ames IA, 50011-3232 } } } } Ph: 515-294-8187 } } FAX: 515-294-4563 } } } } E-Mail: wesaia-at-iastate.edu } } http://www.marl.iastate.edu } } } } ___________________________________________ } } } } We are using the Epson Photo and are very pleased with the results. Cost } } per print is dependent on the type of paper you use. We do proofs on laser } } paper and use injet matte or glossy for better quality prints. Publication } } prints are usually done on a dye sub printer, although the glossy inkjet } } prints look awfully good. I could send samples if you like. } } } } Greg } } } } Gregory W. Erdos, Ph.D. Ph. 352-392-1295 } } Assistant Director, Biotechnology Program } } PO Box 110580 Fax:352-846-0251 } } University of Florida } } Gainesville, FL 32611 } } --
Linkam is based in Surrey, England and their telephone number is +44 (0= )1737 363476
Our local Linkam distributor is Fryer Co. and they can be reached (847)-669-2000 or by email at fryerco-at-ix.netcom.com.
I have also received technical support from a company called Microdevic= es. I don't know their location or if they sell the products but their toll f= ree number is 1-800-331-6786.
Good luck. Our Linkam stage has performed well.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064 847-938-5024 joe.neilly-at-abbott.com =
WARNING! WARNING! DANGER! In case you can not tell from the line below which states that "...we have available...", I have a vested interest in this. I am trying to offer you something for sale (GASP!) which you may be interested in. If you are horrified by advertising, please read no further! (unless you are trying to get a good scare in time for Halloween).
I didn't see the originial inquiry, but let me state that we have available a dye sublimation printer for $2300. Print quality is on par with the $5-9K units, and far better than you will ever see from an inkjet printer. You will get photograph quality prints from the printer. If you are interested in futher details, plese email me or call me using the information below.
Sincerely, J. Haley
****************************** Jim Haley Applications Engineer I-CUBE 2411 Crofton Lane, Suite 14A Crofton, MD 21114 voice: (301) 858-0505 fax: (301) 858-0615 web site: http://www.i-cubeinc.com e-mail: haley-at-i-cubeinc.com ******************************
} } drose-at-wlgore.com-at-Sparc5.Microscopy.Com wrote: } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Hello List, } } } } } } Thanks to all who responded to my posting. All the information has been } } } helpful. Responses are summarized below. } } } } } } David Rose } } } W.L. Gore and Associates } } } } } } =========================================== } } } } } } I've seen posts which responded to the original "HP vs Epson vs Alps" } } } which imply Alps is an ink jet. The original poster may have been } } } referring to the Alps 1300 which is a dual mode 600dpi ink jet and } } } dye-sub printer for ~$500. I was personally impressed with the example } } } Alps sent me, but I also hear it may be the slowest dye-sub on the } } } market ... and we all know printer performance has more to do with how } } } well the printer interfaces with the OS. I know nothing beyond this ... } } } but certainly am curious if someone has practical experience. } } } } } } cheerios, shAf } } } } } } {} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} } } } Michael Shaffer, R.A. - ICQ 210524 } } } Geological Science's Electron Probe Facility - University of Oregon } } } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/ } } } } } } ___________________________________________ } } } } } } Only real photo quality is dye sublimation (kodak, codonics, tektronics, } } } etc) } } } which are expensive (maybe $5K and up). Inkjets like you name, with } } } special, } } } expensive glossy paper and 1200 dpi are almost as good for a few hundred $. } } } I } } } like the Epson Stylus Photo (EX) myself. All will be color (but make good } } } B&W) } } } unless you get a laser printer. Dave Pevear, Houston } } } } } } ___________________________________________________ } } } } } } I can't tell you anything about Alps, but I do use HP inkjets and laserjets } } } and an Epson 600. } } } } } } The HP inkjets to a fair to middling job of either color or black and white } } } (true for all the Deskjet 6XX models I've used - 600, 660). Their fast and } } } cheap and not bad if you're not at all picky. The pictures are grainy. } } } } } } I have an HP Laserjet 5P I use for black and white. At the present time } } } we usually give a polaroid print to the person requesting work and laserjet } } } copies (four to a page which makes the laserjet "prints" just about the } } } same size as the polaroids). The laserjet pictures are not by any stretch } } } of the imagination "photoquality" but they are very good and give more than } } } enough detail for most purposes. } } } } } } The Epson 600 is used the same way as the laserjet for color photos. } } } Again, these are not "photoquality", but they are amazingly good even at } } } 720X720 dpi. However, even at 720 dpi, it is excruciatingly slow. I think } } } Epson got it's fraction turned around when they figured out the page rate. } } } I think they quote something like 2 pages a minute. 2 minutes per page } } } would be far closer to the truth. } } } } } } All of this is on the laserjet paper they use around here for normal office } } } work for the laserjet or good (not the really good shiny stuff) inkjet } } } paper for the Epson. If I started using the really good stuff I suspect } } } the results would be noticably better but so would the cost per page which } } } was the point in going to these printers in the first place. } } } } } } Ive never figured out the actual cost per page for any of these. I'm } } } pretty sure it's far below the $2/print I pay for B&W polaroid or } } } $.75/print for video printers. } } } } } } ============================================================ } } } Bede Willenbring Phone: (651)236-5470 } } } H.B. Fuller Company FAX: (651)236-5430 } } } Research Chemist E-mail: Bede.Willenbring-at-HBFuller.com } } } P.O. Box 64683 } } } St. Paul, MN 55164-0683 } } } ___________________________________________ } } } } } } B&W printer are actually harder to find than good color ones - the Lexmark } } } 5700, Epson Photo ot Cannon 7004 all are great color printers - (6 or 7 } } } colors and } 1000 dpi). } } } } } } Bill Miller } } } ___________________________________________ } } } } } } We have both. } } } HP LaserJet's which we use for viewing B&W images. The 600 dpi output is } } } more than good enough for most work and we seldom go to film anymore. } } } You won't have any trouble getting someone who has one to print a BMP or } } } other photo out to show you how good they look. } } } I also have an Epson 1440x720 dpi color printer for near photo quality } } } in color, and I seldom use it for that, although I just bought a digital } } } camera. I also have a 300 dpi HP 1600color inkjet which I use the heck } } } out of because its fast and good enough for initial work like cropping } } } and framing and positioning. Film is fast becoming sidelined in all the } } } branches of our company, and I see the same thing wherever I go. I am } } } getting a 1200 dpi LaserJet with my next machine so my B&Ws will look } } } even better. The metallurgists are jealous because they are stuck with } } } an expensive, slow photo printing system which uses proprietary file } } } formats. They will be upgraded someday. } } } } } } I don't know anything about Alps. HP is so easy to set up (network too) } } } and has the fastest drivers, Epson had to outdo them big in dpi to get } } } my attention. } } } } } } Tim Chavez } } } Boeing Chem Characterization Lab } } } ___________________________________________ } } } } } } We have a HP 890C Inkjet and you can't beat it. It is has the Kodak } } } PhotoRes processing and really does wonderful output. It is as good as a } } } sublimation dye printer (we have two) when the special Kodak Photo Deluxe } } } paper is used. It is very good on regular paper, better still on the HP } } } premium paper. The printer is about $360. You can get a 722C for about } } } $260 which uses the same cartridges and is a little slower. There are web } } } sites that you can find the cost of the media and ink cartridges. Several } } } people here who have Epson inkjet have been using ours to print critical } } } stuff and slides because the quality is so much better. Don't have } } } experience on the Alps. } } } } } } -Scott } } } } } } Scott D. Walck, Ph.D. } } } PPG Industries, Inc. } } } Guys Run Rd. (packages) } } } P.O. Box 11472 (letters) } } } Pittsburgh, PA 15238-0472 } } } } } } Walck-at-PPG.com } } } } } } (412) 820-8651 (office) } } } (412) 820-8161 (fax) } } } } } } ___________________________________________ } } } } } } We recently purchased a Kodak SP700 dye sub printer. The thing takes a } } } couple minutes per print, and prints only on their specialized paper, but } } } the cost is low per print ( {70 cents) and does a very good gray scale } } } rendition. } } } } } } We use an Epson 600, a Lexmark Optra Rt+ and a HP Laserjet 4M for general } } } purpose printing. it is enough for most people. If they want a real good } } } print we will print on the Kodak or send the image back to our SEM photo } } } CRT (special hardware required). } } } } } } Warren E. Straszheim } } } 23 Town Engineering } } } Iowa State University } } } Ames IA, 50011-3232 } } } } } } Ph: 515-294-8187 } } } FAX: 515-294-4563 } } } } } } E-Mail: wesaia-at-iastate.edu } } } http://www.marl.iastate.edu } } } } } } ___________________________________________ } } } } } } We are using the Epson Photo and are very pleased with the results. Cost } } } per print is dependent on the type of paper you use. We do proofs on laser } } } paper and use injet matte or glossy for better quality prints. Publication } } } prints are usually done on a dye sub printer, although the glossy inkjet } } } prints look awfully good. I could send samples if you like. } } } } } } Greg } } } } } } Gregory W. Erdos, Ph.D. Ph. 352-392-1295 } } } Assistant Director, Biotechnology Program } } } PO Box 110580 Fax:352-846-0251 } } } University of Florida } } } Gainesville, FL 32611 } } } } --
If someone's got the telephone number handy for RMC, Inc., I would greatly appreciate it! None of my current (outdated?) numbers work. Also, if there is a website and/or e-mail information would be appreciated. Attempting to obtain certification of Y2K compliance (mandatory for all of our equipment). Thanks!
Walt Bobrowski Subcellular Pathology Parke-Davis Research Ann Arbor, MI 48105
3450 South Broadmont, Suite 100 Tucson, AZ 85713 Tel. (520) 903-9366 Fax (520) 903-0132
RMC was recently purchased in its entirety by Ventana Medical Systems. I do not know how far along they are in transferring manufacturing and product support, so if the above information doesn't work you should try Ventana at:
3865 North Business Center Drive Tucson, AZ 85705 Tel. (520) 887-2155
RMC used to have a web page at: {A HREF="http://www.rmc- scientific.com/microtomes/"} RMC Home Page {/A} (http://www.rmc-scientific.com/microtomes/) but I haven't tried it in some time.
Good luck, hope this helps.
Cheers,
Bob Chiovetti Microimaging Technologies, Inc. (520) 546-4986 rchiovetti-at-aol.com
I wish to thank all who responded to my question on Spurr's resin. My problem with brittle blocks has been solved. Thanks to all again. -- Best Regards, Gregory Rudomen Technical Specialist University Microscopy Imaging Center State University of New York at Stony Brook 516-444-3126 Greg-at-umic.sunysb.edu ********************************************************** Standard disclaimer: The opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. **********************************************************
I wish to thank all who responded to my question on Spurr's resin. My problem with brittle blocks has been solved. Thanks to all again. -- Regards, Gregory Rudomen Technical Specialist University Microscopy Imaging Center State University of New York at Stony Brook 516-444-3126 Greg-at-umic.sunysb.edu ********************************************************** Standard disclaimer: The opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. **********************************************************
The 1996 Nobel Prize in Physics was shared by Gerd Binning, Heinrich Rohrer, and Ernst Ruska. "Reviews of Modern Physics," Volume 59 (3), July 1987 p. 627 (Part 1) contains the complete text of their addresses on the occasion of the award ( both STM and TEM are covered). There are lots of nice figures, pictures and diagrams of old TEMs and many references to the early literature.
} ---------- } From: Chris McDermott } Sent: Monday, October 26, 1998 7:55 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: TEM development } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I don't know if you answer stuff like this but I'm kind of stuck } at the moment. I'm looking for information on the investigators involved } in the development of the TEM but am unable to find any information } anywhere on the web. If you know of anywhere this information may be } found I would appreciate it a lot. } } Yours, } } Chris McDermott } }
I have relayed all the replies to the Research Center that requested the SEM w/ computerized stage. The researcher has told me that at this time he has enough "resources" for this project to present to his management. He thanks everyone for their quick response.
J. Roy Nelson Material Testing Laboratory Pennington NJ (609) 730-0575 jrnelson-at-nj1.aae.com
We are evaluating 35 mm slide scanners (digitizers) and are considering the Nikon Coolscan III ($900) and Coolscan 2000 ($1,800). The major difference between the two is optical density max of 3.0 and 3.6, respectively. Otherwise resolution is the same.
Anyone familiar with the NIkon line of scanners? Is the price difference worth it for ODmax of 3.0 to 3.6? Dealers are invited to respond to me directly.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Jane et al: The best fixation of lens( dystrophic deer mouse) I have seen was performed at about 40 degrees C using an EM fixative containing the addition of acrolein (about 1%) and DMSO to the glutaraldehyde buffer mixture. Infiltration and embedding in Spurr's would enhance infiltration and embedding. I hope this helps with some experimentation as the exact procedure would vary somewhat with the species. CAUTION Acrolein is very noxious so must be performed in a hood!! bob m
Electron Microscope Facility Manager/ Research Assistant, non-tenure track faculty appointment
Professional electron microscopist with a minimum of 5 years experience to manage the daily activities of the EM Facility at the University of Missouri-Kansas City School of Dentistry. Duties would include active participation in ongoing research, troubleshooting, scheduling, and teaching on facility instruments: Philips CM12 STEM with new Princeton Gamma Tech EDS system; Philips 515 SEM with new Princeton Gamma Tech EDS system and Robinson backscatter electron detector; and recently-installed Philips Environmental SEM equipped with Schottky field emission source, gaseous electron detector and new Princeton Gamma Tech EDS system. Applicant should have operational and maintenance experience with transmission and scanning electron microscopes. Additionally knowledge of energy dispersive spectroscopy and specimen preparatory techniques is required. The position is affiliated with the Department of Oral Biology at UMKC School of Dentistry. Salary is commensurate with level of education and experience. Please respond by E mail to the Chairman, Dr. David Eick "eickd-at-umkc.edu {mailto:eickd-at-umkc.edu} " with resume/CV.
John asks ... } } } We are evaluating 35 mm slide scanners (digitizers) and are } considering the } Nikon Coolscan III ($900) and Coolscan 2000 ($1,800). The } major difference } between the two is optical density max of 3.0 and 3.6, respectively. } Otherwise resolution is the same. } } Anyone familiar with the NIkon line of scanners? } Is the price difference worth it for ODmax of 3.0 to 3.6? }
Being able to handle the extra dynamic range would depend on your applications ... but I'll let you know a typical 35mm slide of, say, an outside scene in sunlight would exceed 3.6, but I'd guess at an indoor natural light slide being less than 3. The other issue is the color depth during the scan process. I'm unfamiliar with these new Coolscans, but do NOT settle for 8bit depth per RGB channel ... you want at least 10bits per channel, and preferably 12bits. ... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I imagine that this was covered in the past, but here is the question = once again. What PC (Intel) based image analysis software is out there? =
My colleague needs to threshold and differentiate a second phase in SEM = acquired TIFF images and to determine its volume fractions. An = "illumination normalization function" (for a lack of a better term) = would be a nice feature to remove, prior to thresholding, the effect of = the drop off in SEM image intensity with distance from the image center.
We do have an old-ish PC based system hooked up to a CCD cameras and = optical microscopes that can read TIFF, but it is limited to working = with images no greater than 640x480 pixels; whereas the SEM ones are = 1x1k or 2x2k. We would like to do this on the cheap (public domain or = shareware), if possible. A demo version of the commercial software = might also be of interest to show the management its utility.
Thanks in advance for any leads,
Valdemar Furdanowicz valdemar-at-fast.net HRL G-165 Bethlehem Steel Co, Bethlehem PA 18016
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} Hello, {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} I imagine that this was covered in = the past, but=20 here is the question once again. What PC (Intel) based image = analysis=20 software is out there? {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} My colleague needs to threshold and=20 differentiate a second phase in SEM acquired TIFF images and to = determine its=20 volume fractions. An "illumination normalization function" = (for a lack=20 of a better term) would be a nice feature to remove, prior to = thresholding, the=20 effect of the drop off in SEM image intensity with distance from the = image=20 center. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} We do have an old-ish PC based = system hooked up=20 to a CCD cameras and optical microscopes that can read TIFF, but it is = limited=20 to working with images no greater than 640x480 pixels; whereas the SEM = ones are=20 1x1k or 2x2k. We would like to do this on the cheap (public domain or=20 shareware), if possible. A demo version of the commercial software = might=20 also be of interest to show the management its utility. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Thanks in advance for any = leads, {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Valdemar Furdanowicz {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {A=20 href=3D"mailto:valdemar-at-fast.net"} valdemar-at-fast.net {/A} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {FONT size=3D2} HRL = G-165 {/FONT} {/DIV} {DIV} {FONT size=3D2} Bethlehem Steel Co, {/FONT} {/DIV} {DIV} {FONT size=3D2} Bethlehem PA 18016 {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {/BODY} {/HTML}
Hi fellow electron microscopists, =20 I've followed with interest the recent discussion on cleaning the window = of an EDS detector, but mine is a 13 y.o. windowless Link (now Oxford) = on a clean vacuum STEM (VG-HB501). (It goes behind a high vacuum valve = when not in use.) I have periodically de-iced it by warming up to room = temperature under high vacuum (mine was built before the era of detector = conditioners), but now my low end is not quite what it used to be and I = think that I've finally built up a noticeable hydrocarbon layer on its = surface. Any ideas on how to clean mine? =20 Valdemar Furdanowicz valdemar-at-fast.net HRL G-165 Bethlehem Steel Co. Bethlehem, PA 18016 =20
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 = HTML//EN"} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} Hi fellow electron = microscopists, {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} I've followed with interest the = recent=20 discussion on cleaning the window of an EDS detector, but mine is a 13 = y.o.=20 windowless Link (now Oxford) on a clean vacuum STEM (VG-HB501). = (It goes=20 behind a high vacuum valve when not in use.) I have periodically = de-iced=20 it by warming up to room temperature under high vacuum (mine was built = before=20 the era of detector conditioners), but now my low end is not quite what = it used=20 to be and I think that I've finally built up a noticeable hydrocarbon = layer on=20 its surface. Any ideas on how to clean mine? {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Valdemar Furdanowicz {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {A=20 href=3D"mailto:valdemar-at-fast.net"} valdemar-at-fast.net {/A} {/FONT} {/DIV} {DIV} {FONT size=3D2} HRL G-165 {/FONT} {/DIV} {DIV} {FONT size=3D2} Bethlehem Steel Co. {BR} Bethlehem, PA = 18016 {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {/BODY} {/HTML}
Jane et al: The best fixation of lens( dystrophic deer mouse) I have seen was performed at about 40 degrees C using an EM fixative containing the addition of acrolein (about 1%) and DMSO to the glutaraldehyde buffer mixture. Infiltration and embedding in Spurr's would enhance infiltration and embedding. I hope this helps with some experimentation as the exact procedure would vary somewhat with the species. CAUTION Acrolein is very noxious so must be performed in a hood!! bob m
To: Chris McDermott For historical info try "Early History of the Electron Microscope" by L. Marton, San Francisco Press, Inc., CA, 1968, Lib. Con. Cat. # 68-19314 and "Electron Microscopy 1978, Vol. III: State of the Art Symposia", Papers presented in Symposia at the Ninth International Congress on Electron Microscopy held in Toronto, Canada 1978, edited by J.M Sturgess. It was published by the Microscopical Society of Canada, ISBN 0 920622 08 9. Bob Santoianni Emory University Hospital robert_santoianni-at-emory.org
Yes, I do have a suggestion--talk to the manufacturer. Si(Li)s are very troublesome creatures, you don't want to damage it!
And I am very impressed that it took 13 years to gum it up. When you say you have a clean system you mean it :)
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
We use a Polaroid 35mm slide scanner, I forget the model, but we got it with a special device that permits scanning of entire MICROSCOPE SLIDES at 27000 dpi! You can also do 35mm slides. It gives a great low-power image that can be magnified digitally without loss. The product, Pathscan, is made by Meyer Instruments {http://www.meyerinst.com/html/eureka/default.html} . No, I am not a vendor of anything other than the good news of this product! Dave Pevear, Houston.
John J. Bozzola wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are evaluating 35 mm slide scanners (digitizers) and are considering the } Nikon Coolscan III ($900) and Coolscan 2000 ($1,800). The major difference } between the two is optical density max of 3.0 and 3.6, respectively. } Otherwise resolution is the same. } } Anyone familiar with the NIkon line of scanners? } Is the price difference worth it for ODmax of 3.0 to 3.6? } Dealers are invited to respond to me directly. } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ####################################################################
Dr. Dinesh Srivastava Research Fellow IRC for Materials of High Performance University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK Phone (O) 0121-414-3447 Fax (O) 0121-414-3441 (Res.) 0121-472-3298 Email:D.SRIVASTAVA-at-bham.ac.uk
I have some very important blocks of human duodenum which have been processed through to Araldite CY212. Unfortunately the embedding is very poor. The resin making up the bulk of the block is fine but in and around the tissue the resin is 'gooey' and sections float apart on flattening. Does anyone have a tried and trusted method for removing the existing resin and re-embedding with fresh? If so I would be eternally grateful if you could pass it on to me as quickly as possible.
Yours hopefully
Barry Shaw (Snr. EM Technician, Univ. Hospital Medical School Anatomy Dept., Nottingham, UK)
In a message dated 98-10-26 16:50:23 EST, valdemar-at-fast.net writes:
{ { We do have an old-ish PC based system hooked up to a CCD cameras and optical microscopes that can read TIFF, but it is limited to working with images no greater than 640x480 pixels; whereas the SEM ones are 1x1k or 2x2k. We would like to do this on the cheap (public domain or shareware), if possible. A demo version of the commercial software might also be of interest to show the management its utility. } }
Hi Valdemar,
There are certainly some high-powered (translation: pricey) image analysis software packages out there. Feel free to contact me off-list if you need additional information.
You might first want to try a PC-based variation of the great public domain software, NIH Image. NIH Image is of course for Macs, but there is now a Windows version. The name is Scion Image, and it comes in a Windows 95 and a Windows NT version. The download page does not mention Windows 98.
I haven't heard from anyone who has tried it, but it apparently has full NIH Image functionality, only it runs on PC platforms. From what you say about your computer, you may be short some horsepower for the analysis routines, but it would certainly be worth a try. One thing is certain, you can't beat the price...it's free. Plus, you also get free technical support when you register to download.
Scion Image is ported to the Scion Corporation's website. You can read about Scion Image here: {A HREF="http://www.pathsoc.org.uk/wwwboard/messages/20.html"} NIH Image analysis software now out for Windows {/A} (http://www.pathsoc.org.uk/wwwboard/messages/20.html)
And you can download it from: {A HREF="http://www.scioncorp.com/pages/menu.htm"} Scion Corporation {/A} (http://www.scioncorp.com/pages/menu.htm)
**Disclaimer:** I have no financial interest or business relationships with Scion Corporation. This information is just offered as a possible solution to your problem.
Let us know how things turn out if you decide to try the software.
Cheers, Bob
Robert (Bob) Chiovetti Microimaging Technologies, Inc. Tucson, AZ Tel / Fax (520) 546-4986 rchiovetti-at-aol.com
thee is also a good book by zworkin(spelling?) on electron mic. and electron optics that will give a picture of some of the early work.... I belive this is from the late 40's
Edward Sharpe ( I am not in front of my stash so the title and spelling is the best I can remember....)
BARRY SHAW wrote: } } HELP !!! } } I have some very important blocks of human duodenum which have been } processed through to Araldite CY212. Unfortunately the embedding is } very poor. The resin making up the bulk of the block is fine but in } and around the tissue the resin is 'gooey' and sections float apart } on flattening. Does anyone have a tried and trusted method for } removing the existing resin and re-embedding with fresh? } If so I would be eternally grateful if you could pass it on to me } as quickly as possible. }
Barry,
Long ago, I had a problem with human kidney embedded in Epon-Araldite that was similar to what you describe. I cooked the blocks overnight at 100 C and that "cured" the problem. I don't know of a way to remove gooey resin without compromising the tissue.
I've had this problem before, but with Spurrs rather than Araldite. I had limited success by using propylene oxide to remove the gooey stuff.
First of all, have you tried cranking up the heat in your polymerizing oven? Sometimes you can rescue the blocks by really "cooking" them at about 70-75 degrees C for 24 hours. If this doesn't work, proceed to plan B...
Try using a dissecting scope and the trimming block on your microtome to remove as much of the polymerized resin as possible with razor blades, then cut under the specimen to remove it from the block. Careful here, if you use too much force the valued specimen will go flying across the room never to be seen again.
It depends on how much crosslinking has taken place in the soft parts, but you should be able to remove most of it with propylene oxide. Place the unblocked specimens in small vials with a large excess of propylene oxide, and place the vials on a rotator or agitator. Make lots of changes of propylene oxide over a period of 24-48 hours. From that point you should be able to infiltrate as usual. Try an ascending series of resin concentrations that are cut with propylene oxide, then pure resin overnight. During the infiltration use a bell jar or a vacuum chamber if you have access to one, and a vacuum oven for polymerization is also a good idea.
My experience has been that unblocked and reinfiltrated specimens are never as good as they should be, but reprocessing usually makes the specimens better than what came out of the oven the first time around!
Good luck, hope this helps!
Cheers,
Bob
Bob Chiovetti Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax 520-546-4986 rchiovetti-at-aol.com
Our group currently has a JEOL FX 2000 TEM available for sale to anyone interested. The system includes a cold and hot stage, PC driven X-ray system, and STEM attachment. Please respond if interested.
M.W. Rigler, Ph.D. MAS, Inc. Suwanee GA 770-866-3218
I have purchased two SEMs, transported them across country, and am preparing to set up a lab. Aside from the insurance on the building, I will need to insure the equipment inside, hence the question: What is the value of the microscopes? There is of course the purchase price (in my case, $ 3,800 and $ 2,500, respectively, for a JEOL 35 and Philips 500). Once in place and running, they are obviously going to be worth a whole lot more, but how much more???? I'm sure my insurance agent will be of no help, as there is no 'blue book' values for used SEMs. Any suggestions are most welcome! Thank you. Thank you. Thank you.
A collegegue of mine has tungsten carbide particles and was wondering if they can be microtomed....i suppose it's possible but i'm not familiar enough with the conditions to use, i.e. resin, cutting speed, etc...i have a 45 deg diamond knife and i've been using spurrs for my other samples
if anyone has any suggestions or has some experience in cutting metal/caramic particles...i'd appreciate any information
thanks in advance
Michael Mandanas Particulate Materials Center Penn State University University Park, PA 16802 email: mxm67-at-email.psu.edu
Valdemar, I would agree with Bob Chiovetti's comments about Scion image - it is very good value for (no) money. However, it has suffered rather in the translation from Mac to PC. NIH-Image on a Mac is a perfectly robust, bug-free program with lots of useful features. Scion image on a PC works, but it is fairly easy to crash (e.g. if you ask it to outline any particles it's counting) and the 0 and 255 levels are reversed in comparison with other PC TIFF images. Also the TIFF file format it uses can't be read by other PC programs. I use it in conjunction with Adobe Photoshop and can do everything I need to (apart from do decent 3D plots of AFM images - the Mac spinoffs such as Image SXM have no counterpart on the PC). Nevertheless, I think all credit is due to Scion for making the product available on a PC, for nothing. The price of most image analysis software is frightening! I have no idea whether new versions are planned or whether the bugs will be fixed - I'm just an almost satisfied customer.
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK
e-mail richard.beanland-at-gecm.com
} ---------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------. } } } In a message dated 98-10-26 16:50:23 EST, valdemar-at-fast.net writes: } } { { We do have an old-ish PC based system hooked up to a CCD cameras and } optical microscopes that can read TIFF, but it is limited to working with } images no greater than 640x480 pixels; whereas the SEM ones are 1x1k or 2x2k. } We would like to do this on the cheap (public domain or shareware), if } possible. A demo version of the commercial software might also be of interest } to show the management its utility. } } } } } Hi Valdemar, } } There are certainly some high-powered (translation: pricey) image analysis } software packages out there. Feel free to contact me off-list if you need } additional information. } } You might first want to try a PC-based variation of the great public domain } software, NIH Image. NIH Image is of course for Macs, but there is now a } Windows version. The name is Scion Image, and it comes in a Windows 95 and a } Windows NT version. The download page does not mention Windows 98. } } I haven't heard from anyone who has tried it, but it apparently has full NIH } Image functionality, only it runs on PC platforms. From what you say about } your computer, you may be short some horsepower for the analysis routines, but } it would certainly be worth a try. One thing is certain, you can't beat the } price...it's free. Plus, you also get free technical support when you } register to download. } } Scion Image is ported to the Scion Corporation's website. You can read about } Scion Image here: } {A HREF="http://www.pathsoc.org.uk/wwwboard/messages/20.html"} NIH Image } analysis software now out for Windows {/A} } (http://www.pathsoc.org.uk/wwwboard/messages/20.html) } } And you can download it from: } {A HREF="http://www.scioncorp.com/pages/menu.htm"} Scion Corporation {/A} } (http://www.scioncorp.com/pages/menu.htm) } } **Disclaimer:** I have no financial interest or business relationships with } Scion Corporation. This information is just offered as a possible solution to } your problem. } } Let us know how things turn out if you decide to try the software. } } Cheers, } Bob } } Robert (Bob) Chiovetti } Microimaging Technologies, Inc. } Tucson, AZ } Tel / Fax (520) 546-4986 } rchiovetti-at-aol.com
You would need first to check the relative hardness of diamond and tungsten carbide; I think diamond will win by a short straw. You may have a problem with the very hard tungsten carbide embedded in softer resin and the passing knife edge may just rip the W-carbide out of the resin matrix. Maybe a better approach would be to embedd the material in resin and then etch a small portion of the resin away using say sodium methoxide, wash well with EtOH and look (and analyse ?) in a high resolutio FEG-SEM.
Patrick Echlin Multi-Imaging Centre Cambridge UK
On Tue, 27 Oct 1998, Michael P. Mandanas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all } } A collegegue of mine has tungsten carbide particles and was wondering if } they can be microtomed....i suppose it's possible but i'm not familiar } enough with the conditions to use, i.e. resin, cutting speed, etc...i have } a 45 deg diamond knife and i've been using spurrs for my other samples } } if anyone has any suggestions or has some experience in cutting } metal/caramic particles...i'd appreciate any information } } thanks in advance } } Michael Mandanas } Particulate Materials Center } Penn State University } University Park, PA 16802 } email: mxm67-at-email.psu.edu } } } }
} Valdemar, } I would agree with Bob Chiovetti's comments about Scion image - it is } very good value for (no) money. However, it has suffered rather in the } translation from Mac to PC. NIH-Image on a Mac is a perfectly robust, bug-free } program with lots of useful features. Scion image on a PC works, but it is } fairly easy to crash (e.g. if you ask it to outline any particles it's } counting) and the 0 and 255 levels are reversed in comparison with other PC } TIFF images. Also the TIFF file format it uses can't be read by other PC } programs. I use it in conjunction with Adobe Photoshop and can do everything I } need to (apart from do decent 3D plots of AFM images - the Mac spinoffs such as } Image SXM have no counterpart on the PC).
And then their is Imagetool (Win95). I haven't used it enough to comment on any peculiarities with TIFF files in particular, but it is also free, via:
http://ddsdx.uthscsa.edu/dig/itdesc.html
Usual disclaimers apply,
Keith -- Dr. Keith R. Hallam University of Bristol, Interface Analysis Centre, Oldbury House, 121, St. Michael's Hill, Bristol, BS2 8BS, England Telephone: + 44 (0)117 925 5666 | E-mail: k.r.hallam-at-bristol.ac.uk Facsimile: + 44 (0)117 925 5646 | URL: http://zeus.bris.ac.uk/~phkrh/
I guess I would ask myself how much I could afford if there were to be major damage to the instruments ( fire, etc) . If replacing the instruments with my own money represented an "affordable" impact, then I would not consider putting much money into the insurance. I would rather spend that money in supplies and or service contracts.
BTW, I 'm not sure I agree with your assessment that the instruments will be worth a lot more once they are operating. Of course this will depend somewhat on the present condition and how much you had to invest to get them back into operation.
My thoughts. I am curious to see what others think.
Jordi Marti
I have purchased two SEMs, transported them across country, and am preparing to set up a lab. Aside from the insurance on the building, I will need to insure the equipment inside, hence the question: What is the value of the microscopes? There is of course the purchase price (in my case, $ 3,800 and $ 2,500, respectively, for a JEOL 35 and Philips 500). Once in place and running, they are obviously going to be worth a whole lot more, but how much more???? I'm sure my insurance agent will be of no help, as there is no 'blue book' values for used SEMs. Any suggestions are most welcome! Thank you. Thank you. Thank you.
I have been working with a number of equipment brokers in the resale of SEMs for about five years. To give you an idea of the value, a JEOL 840 installed and warranteed for one year goes for about 40K. The same SEM is "wholesaled" (trade in value) by JEOL for 20K.
There is a lot of "added value" to any SEM as labor costs are fixed. It costs about 4K to dis-assemble and re-assemble most SEMs. Moving adds about 3K. Warranty adds about 2k per quarter. This all adds to the SEM value even though the initial cost is low. In my opinion, the replacement value for a JEOL 35C is anywhere from 10K to 15K. For insurance purposes it doesn't matter what you paid for the equipment, it is the cost to replace it.
For higher end equipment (CD measurements systems), the market is somewhat "skewed" at this time as there is a glut of equipment especially from Korea and Japan. The Asia flu is causing them to shut many semi-conductor facilities down. Supply and demand still dictate the market. A CD measurement systems that sold for average 1 million new generally resale for about 200k after five years (Hitachi S-6000). the same equipment is selling for about 50K, a seventy five percent reduction.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Greetings esteemed microscopy mentors, } } I have purchased two SEMs, transported them across country, and am preparing } to set up a lab. Aside from the insurance on the building, I will need to } insure the equipment inside, hence the question: What is the value of the } microscopes? } There is of course the purchase price (in my case, $ 3,800 and $ 2,500, } respectively, for a JEOL 35 and Philips 500). Once in place and running, they } are obviously going to be worth a whole lot more, but how much more???? I'm } sure my insurance agent will be of no help, as there is no 'blue book' values } for used SEMs. Any suggestions are most welcome! Thank you. Thank you. Thank } you. } } Paul Grover } pbgrover-at-aol.com
} Dear Paul, } } I have been working with a number of equipment brokers in the resale of SEMs for } about five years. To give you an idea of the value, a JEOL 840 installed and } warranteed for one year goes for about 40K. The same SEM is "wholesaled" (trade in } value) by JEOL for 20K. } } There is a lot of "added value" to any SEM as labor costs are fixed. It costs } about 4K to dis-assemble and re-assemble most SEMs. Moving adds about 3K. Warranty } adds about 2k per quarter. This all adds to the SEM value even though the initial } cost is low. In my opinion, the replacement value for a JEOL 35C is anywhere from } 10K to 15K. For insurance purposes it doesn't matter what you paid for the } equipment, it is the cost to replace it. } } For higher end equipment (CD measurements systems), the market is somewhat } "skewed" at this time as there is a glut of equipment especially from Korea and } Japan. The Asia flu is causing them to shut many semi-conductor facilities down. } Supply and demand still dictate the market. A CD measurement systems that sold for } average 1 million new generally resale for about 200k after five years (Hitachi } S-6000). the same equipment is selling for about 50K, a seventy five percent } reduction. } } Hope this helps. } } Earl Weltmer
} } } Pbgrover-at-aol.com-at-sparc5.microscopy.com wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Greetings esteemed microscopy mentors, } } } } I have purchased two SEMs, transported them across country, and am preparing } } to set up a lab. Aside from the insurance on the building, I will need to } } insure the equipment inside, hence the question: What is the value of the } } microscopes? } } There is of course the purchase price (in my case, $ 3,800 and $ 2,500, } } respectively, for a JEOL 35 and Philips 500). Once in place and running, they } } are obviously going to be worth a whole lot more, but how much more???? I'm } } sure my insurance agent will be of no help, as there is no 'blue book' values } } for used SEMs. Any suggestions are most welcome! Thank you. Thank you. Thank } } you. } } } } Paul Grover } } pbgrover-at-aol.com
Leica manufactures a microtome that can accommodate a micromilling tool bit. This machine is a combination of a microtome and a macro-milling machine that you might use in the machine shop. It enables production of mirror-like metallic surfaces and is likely appropriate for your application. An example of its use to reconstruct 3-D images of Pb-Sn alloys can be found in Acta Mater. 45 2279 (1997).
I am in no way affiliated with Leica.
Hope this helps.
Regards, Tracey Wolfsdorf Brenner
Disclaimer: The comments above do not represent those of Intel Corporation.
} -----Original Message----- } From: Michael P. Mandanas [SMTP:mxm67-at-email.psu.edu] } Sent: Tuesday, October 27, 1998 10:00 PM } To: microscopy-at-sparc5.microscopy.com } Subject: sectioning metals? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all } } A collegegue of mine has tungsten carbide particles and was wondering if } they can be microtomed....i suppose it's possible but i'm not familiar } enough with the conditions to use, i.e. resin, cutting speed, etc...i have } a 45 deg diamond knife and i've been using spurrs for my other samples } } if anyone has any suggestions or has some experience in cutting } metal/caramic particles...i'd appreciate any information } } thanks in advance } } Michael Mandanas } Particulate Materials Center } Penn State University } University Park, PA 16802 } email: mxm67-at-email.psu.edu } }
Hi everyone, My dissecting scope and computer are in different rooms so I would like to buy a digital camera that allows image preview and image storage on removable media for transfer to the computer. Olympus manufacture a digital camera system (DP10) which fits the bill. Resolution is listed as 1280x1024. If I recall the thread on resolution correctly, this should be sufficient for output similar to that that I would achieve via film, am I right? Any users out there with thoughts on this system? Cheerio, John.
________________________ C. John Runions, Ph.D. Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
Dear All: We have a JEOL 100CX-II transmission microscope, which we would like to get rid off. Could any of you give me some idea as to the value of this scope? It is in excellent working condition. Thanks in advance for your opinion. Yuhui Xu, MD EM Core,DFCI
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I am looking for the email addresses for Hans Cerva and Oliver Eibl at Siemens. Any help would be appreciated. Thanks. Roseann -- Dr. Roseann Csencsits Electron Microscopy Center Building 212/C217 Argonne National Laboratory 9700 South Cass Avenue Argonne, IL 60439-4838 Phone: (630) 252-4977 Fax: (630) 252-4798
You actually got some very good prices on your equipment. There is a growing secondary market in used SEMs that provides a reasonable replacement value. There are a number of used equipment brokers out there that would charge 15K and 10K, respectively, for your instruments. If you were able to bypass them, you could probably get those instruments for 2/3 their asking price.
For insurance values, you should really use those broker figures, since you have no assurance that you would be able to replace them at anything near the prices you paid. The broker purchase prices can be verified by calling the manufacturer's sales reps and asking them what would be a reasonable asking price and then adding 50%. The brokers generally get their instruments through trade-ins arranged through the manufacturers. In the chain of things, the manufacturer will get a mark-up on the instrument, as will the broker. You obviously managed to bypass both of them in your purchases.
Also add at least a couple of thousand for the inspection and installation of a used SEM. You definitely want to ensure that it works before you buy and will want to have it installed by someone who can give you some assurance that it will operate to your needs. If you feel that you have that capability, then you can save that much more.
} Greetings esteemed microscopy mentors, } } I have purchased two SEMs, transported them across country, and am } preparing to set up a lab. Aside from the insurance on the } building, I will need to insure the equipment inside, hence the } question: What is the value of the microscopes? There is of course } the purchase price (in my case, $ 3,800 and $ 2,500, respectively, } for a JEOL 35 and Philips 500). Once in place and running, they are } obviously going to be worth a whole lot more, but how much more???? } I'm sure my insurance agent will be of no help, as there is no 'blue } book' values for used SEMs. Any suggestions are most welcome! } Thank you. Thank you. Thank you. } } Paul Grover } pbgrover-at-aol.com } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
I agree with Patrick - another approach might be your best bet. A few years back, Helmut Gnaegi of Diatome (a diamond knife producer) succeeded in sectioning monolithic tungsten carbide for our laboratory, but the sectioning was a) challenging, and b) the TEM images weren't pretty. Experienced materials science ultramicrotomists like Helmut can and have sectioned everything from silicon wafers to diamond films. (See Microscopy Research and Techniques, vol. 31, p. 308, 1995, for Phil Swab's excellent work on the latter).
The point is, one should push a technique to the extreme only if that technique's attributes best match the experimental needs. Your friend should clearly state his microanalytical expectations, then you and he should work out the appropriate technique. FEG-SEM is a powerful tool that can substitute for many (but certainly not all) TEM materials studies. If TEM is needed, I would first check out the more traditional options of low angle ion thinning or tripod polishing. Failing that, track down the nearest FIB system, swallow hard, and pay what seems like a large amount for a high-end specimen that works first time. IBM Analytical (a frequent contributor to this Listserver and originators of the tripod technique) and Fibics (a Canadian company located in this laboratory) are two providers of FIB service for general materials science.
Use ultramicrotomy only as a last resort. With care, you won't ruin the knife (Helmut and Phil get their best 'hard' materials sections with a 35 degree knife!), but it will require an ultrafine block face of only a few microns, very low cutting speed (like 0.1mm/sec) and, yes, a strong likelihood that the particles will pull out of the resin. Particle size will play a key role in the latter. Micronish will likely be hopeless, but at tens or hundreds of microns, you may 'microfracture' your way through. Finding something that enhances resin cohesion doesn't hurt, like Phil does with a silane from Dow for his work on glasses and semiconductors. Cheers,
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 623-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: Dr P. Echlin[SMTP:pe13-at-cus.cam.ac.uk] } Sent: October 28, 1998 4:10 AM } To: Michael P. Mandanas } Cc: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: sectioning metals? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Michael: } } You would need first to check the relative hardness of diamond and } tungsten carbide; I think diamond will win by a short straw. You may } have a problem with the very hard tungsten carbide embedded in softer } resin and the passing knife edge may just rip the W-carbide out of the } resin matrix. Maybe a better approach would be to embedd the material in } resin and then etch a small portion of the resin away using say sodium } methoxide, wash well with EtOH and look (and analyse ?) in a high } resolutio FEG-SEM. } } Patrick Echlin } Multi-Imaging Centre } Cambridge UK } } On Tue, 27 Oct 1998, Michael P. Mandanas wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi all } } } } A collegegue of mine has tungsten carbide particles and was wondering if } } they can be microtomed....i suppose it's possible but i'm not familiar } } enough with the conditions to use, i.e. resin, cutting speed, etc...i } have } } a 45 deg diamond knife and i've been using spurrs for my other samples } } } } if anyone has any suggestions or has some experience in cutting } } metal/caramic particles...i'd appreciate any information } } } } thanks in advance } } } } Michael Mandanas } } Particulate Materials Center } } Penn State University } } University Park, PA 16802 } } email: mxm67-at-email.psu.edu } } } } } } } } } }
I am working with the ultrastruture of anthers of Ilex paraguariensis. However, during the dehydration of the samples, fixed in a mixture of glutaraldehyde 2,5% and formaldehyde 2%, the anthers shrink, mainly after the secondary fixation with osmium tetroxide. I used acetone or ethanol , in steps of 30, 50, 70, 90, 90, 100, 100, with 15 minutes in each step, at room temperature. The anthers has a very reduced dimension (about 1 mm length) and shrink in the step 100. What to do? Should I to use a low temperature (4oC)? Thanks in advance.
How big are the particles? I worked with WC sub-micron particles and found that the best way was to disperse them on a grid and image them. I assume your particles are bigger.
1. If they are bound with a metal such as Co, you can electron discharge machine a 3 mm disk, then mount and polish it using diamond wheels, and then dimple and ion mill the samples to make electron transparent material.
2. You could mount the particles (if they are powders of WC) in a hard epoxy (Gatan G1?) and try the above steps.
3. Another way to make the samples would be to bind them with a low melting eutectic mixture (netters- any suggestions?), then section them and proceed as above.
Microtoming will probably damage the knife since these particles are extremely hard.
Good luck,
Mohan Kalyanaraman Sr. Staff Material Scientist Mobil Technology Company Paulsboro, NJ 08066
"Michael P. Mandanas" {mxm67-at-email.psu.edu} on 10/27/98 10:00:14 PM
To: microscopy-at-sparc5.microscopy.com cc: (bcc: Mohan Kalyanaraman/EastCoast/Mobil-Notes)
Hi all
A collegegue of mine has tungsten carbide particles and was wondering if they can be microtomed....i suppose it's possible but i'm not familiar enough with the conditions to use, i.e. resin, cutting speed, etc...i have a 45 deg diamond knife and i've been using spurrs for my other samples
if anyone has any suggestions or has some experience in cutting metal/caramic particles...i'd appreciate any information
thanks in advance
Michael Mandanas Particulate Materials Center Penn State University University Park, PA 16802 email: mxm67-at-email.psu.edu
I think you need to check the strength of the fixative buffer. In any event, it is much better to cut the anthers in half across their long axis immediately after they are immersed in the primary fixative to allow the fixative into the anther locule. ASlso, fix for a long time in thye primary fixative
Patrick Echlin Cambridge UK
On Wed, 28 Oct 1998, Rinaldo Pires dos Santos wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear colleagues } } I am working with the ultrastruture of anthers of Ilex paraguariensis. } However, during the dehydration of the samples, fixed in a mixture of glutaraldehyde 2,5% } and formaldehyde 2%, the anthers shrink, mainly after the secondary fixation with osmium } tetroxide. I used acetone or ethanol , in steps of 30, 50, 70, 90, 90, 100, 100, } with 15 minutes in each step, at room temperature. The anthers has a very reduced } dimension (about 1 mm length) and shrink in the step 100. } What to do? Should I to use a low temperature (4oC)? } Thanks in advance. } }
Our group currently has a JEOL FX 2000 TEM available for sale to anyone interested. The system includes a cold and hot stage, PC driven X-ray system, and STEM attachment. Please respond if interested.
M.W. Rigler, Ph.D. MAS, Inc. Suwanee GA 770-866-3218
Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: from rly-za05.mx.aol.com (rly-za05.mail.aol.com [172.31.36.101]) by air-za02.mail.aol.com (v51.9) with SMTP; Tue, 27 Oct 1998 17:13:27 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by rly-za05.mx.aol.com (8.8.8/8.8.5/AOL-4.0.0) with SMTP id RAA01671; Tue, 27 Oct 1998 17:12:14 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id PAA12370 for dist-Microscopy; Tue, 27 Oct 1998 15:40:58 -0600 Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id PAA12353 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 27 Oct 1998 15:40:27 -0600 } From: "Mriglermas-at-aol.com"-at-sparc5.microscopy.com Received: from imo20.mx.aol.com (imo20.mx.aol.com [198.81.17.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id PAA12346 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 27 Oct 1998 15:40:14 -0600 Received: from Mriglermas-at-aol.com by imo20.mx.aol.com (IMOv16.10) id NZKYa03785 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 27 Oct 1998 16:50:09 -0500 (EST) Message-ID: {538306e.36364011-at-aol.com}
Our group currently has a JEOL FX 2000 TEM available for sale to anyone interested. The system includes a cold and hot stage, PC driven X-ray system, and STEM attachment. Please respond if interested.
M.W. Rigler, Ph.D. MAS, Inc. Suwanee GA 770-866-3218
Dear Rinaldo, my experience with fixation of flowers has been that anthers are relatively difficult to fix and embed (especially when they are approching maturity) even when other organs and tissues are well preserved. If adjusting buffer strength and cutting them transversely as Dr. Echlin suggests doesn't do the trick, my suggestion would be that you try much longer dehydration times. I suggest at least an hour at each dehydration step and perhaps leave them overnight at the 70% ethanol stage. Of course the trade-off might be that you extract some of the stuff you wish to preserve. You will have to adjust your protocol on a species to species basis. John
________________________ C. John Runions, Ph.D. Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
} From: Mriglermas-at-aol.com Return-path: {Mriglermas-at-aol.com} To: Microscopy-at-sparc5.microscopy.com
Dear Rinaldo, my experience with fixation of flowers has been that anthers are relatively difficult to fix and embed (especially when they are approching maturity) even when other organs and tissues are well preserved. If adjusting buffer strength and cutting them transversely as Dr. Echlin suggests doesn't do the trick, my suggestion would be that you try much longer dehydration times. I suggest at least an hour at each dehydration step and perhaps leave them overnight at the 70% ethanol stage. Of course the trade-off might be that you extract some of the stuff you wish to preserve. You will have to adjust your protocol on a species to species basis. John
________________________ C. John Runions, Ph.D. Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
Dear Colleagues Being aware of your interest in 3-D reconstruction and visualization of biological object structures, I wonder if you deal with the problem of 3-D epithelial sheets topology reconstruction? If it is so can you send me your papers on this subject? And do you know anybody who work in this field? My own interest is the 3-D histoarchitecture of epithelial sheets in normal development and pathology. In this connect I am addressing you with the following message. As you know, all current attempts to reconstruct a 3-D structure of biological tissues using a serial sections encounter with a typical difficulty. It is that tissue deformation and cell proliferation make a cells shape variable, diverse their adjacency and give a strongly "noised" pictures. So the results obtained give no way to understanding tissue topology and do not permit to deduce the law of 3-D tissue organization.
To overcome the difficulties we have developed the new approach to 3-D reconstruction of cell sheet structures. It is based on the pioneering concept of tissues modular structure and consists of derivation of topological and geometrical models of epithelial spatial organization and their experimental verification. Contrary to the usual tissue reconstruction on a set of serial sections this approach allows to use their minimum and to carry out a more exact restoration of 3-D epithelial structure with less money and time. The approach has enabled us to get the data priority on the laws of spatial organization of simple, pseudostratified and stratified epithelia, as well as to predict and find several earlier unknown topological variants of their histoarchitectonics. The approach has also allowed to describe such new properties of epithelial layers as translation symmetry and stoichiometry.
The results makes a basis for structural histology as a addition to modern structural biology. The approach allow to investigate the more complex topology variants of pseudostratified and stratified epithelia, to find a set of new informative tissue characteristics suitable for diagnostic and also to give the opportunity to predict the changes of tissues in normal development and their transformation in pathology, particularly in malignant growth.
If it is interesting for you, I am ready to consider the possibility of our cooperation.
Yours sincerely, Dr. G.A. Savostyanov. -- | E-mail: savost-at-ief.spb.su | Gennady A. Savostyanov | | Fax +7 (812) 5523012 |Sechenov Inst. of Evolutionary Physiology and | | office +7 (812) 5523090 | Biochemictry Russian Academy of Science, | | | home +7 (812) 5100052 |44 Thores av., 194223, St.Petersburg, Russia |
Can anyone suggest how to cut into a glass pharmaceutical vial with minimal damage and contamination in order to examine an organic film inside it? Presumably some sort of diamond saw. I need a commercial lab who can provide the service, ASAP of course. Hatchet is last resort. Thanks for your help.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
A colleague wants to anesthesize and immobilize whole Drosophila larva on a microscope slide for confocal observation. Any suggestions or references?
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
We used a ion-etching machine without a nitrogen-cooled stage as the last step to prepare thin-foil specimens for a long time. Now we want to have ion mill that have a cooled stage in order to reduce the formation of large amount of amorphous. Could anybody give us some suggestion on where we can find the ion mill and how much it is? Any information and suggestion about it are greatly appreciated.
Jinsong wu LSG2M, Ecole des Mines de Nancy Parc de Saurupt F-54042, Nancy Cedex France
People in our lab have tried a variety of techniques but find that simply putting the larvae in a drop of glycerol and squashing them with the coverslip works best. This does not work if you are interested in the gut because the gut is forced out of the body. The remaining body is like a flat tire. If you want the larvae to survive the problem is more complicated. Try cold temperature (12 C) or mix an anesthetic or muscle relaxant with the food. If you have success with the later post the details. Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
We are interested in using the SEM to measure the concentration of flyash in the atmosphere that has come from coal-fired power plants. One of the issues involved is being able to distinguish particles from power plants from particles originating at other sources. We would appreciate any information people might have on the identification of flyash and determination of its origin. Everett Ramer Federal Energy Technology Center
I am attempting to help a colleague who is in a bind. She needs to get QUICKLY - which is to say - borrow, loan, rent, or buy - a computer controlled stage for an Olympus BH-2 optical microscope.
We're talking to Olympus of course but still ...
Anyone have ideas or a stage available???
Richard Shalvoy rbshalvoy-at-corp.olin.com 203-271-4272
In a message dated 98-10-29 16:55:21 EST, rbshalvoy-at-corp.olin.com writes:
{ { I am attempting to help a colleague who is in a bind. She needs to get QUICKLY - which is to say - borrow, loan, rent, or buy - a computer controlled stage for an Olympus BH-2 optical microscope.
We're talking to Olympus of course but still ...
Anyone have ideas or a stage available???
Richard Shalvoy rbshalvoy-at-corp.olin.com 203-271-4272 } }
Hi Richard,
I recommend that you call Semprex Corporation. They make a complete line of microscope stages to fit just about every microscope: Manual stages, micrometer and leadscrew measuring stages, motorized stages (programmable and non-programmable), hard disk stages, digital readout devices and software, motor controllers and software, etc. They have a dealer network, so there should be someone near you who can respond quickly.
You can reach them at: Semprex Corp. 782 Camden Avenue Campbell, CA 95008 USA Tel. (408) 379-3230 Fax (408) 374-1843
***Disclaimer*** I have no financial interest or business relationships with Semprex, but I have several customers who are pleased with their products.
Hope this helps!
Cheers, Bob *************************** Bob Chiovetti Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com ****************************
This is a multi-part message in MIME format. --------------87761FD3C5399E2310A98724 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Dear Mr. Corwin,
1) Encapsulate the sample fully in Epoxy 2) Using a high speed diamond saw, cut the sample with a RESIN Bonded diamond blade to minimize chipping of the edge. 3) If the cut surface is not good enough, a quick polish maybe necessary.
Try HiRel Laboratory in Spokane, Washington, or Metals Technology in Northridge, Ca. Either of them can be found in directory assistance.
Good Luck,
Gary Liechty Allied High Tech Products, Inc. Products for Metallographic, SEM and TEM Sample Preparation 800-675-1118
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Can anyone suggest how to cut into a glass pharmaceutical vial with } minimal damage and contamination in order to examine an organic film } inside it? Presumably some sort of diamond saw. I need a commercial } lab who can provide the service, ASAP of course. Hatchet is last } resort. Thanks for your help. } } } Leonard Corwin } Research Chemist } Fort Dodge Animal Health } Princeton, NJ 08543-0400
--------------87761FD3C5399E2310A98724 Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Gary Liechty Content-Disposition: attachment; filename="vcard.vcf"
We offer an ion mill that may be of interest to you. The IV3 has a liqui= d nitrogen cold stage as you requested, but it also offers some very new technology in the form of a low energy ion gun. This ion gun operates in=
the range of 100V to 2 kV while still maintaining a relatively high milli= ng rate. It is typically used as a final thinning step after primary thinni= ng with the standard teletwin high energy gun. As you mentioned reducing amorphous damage, the low energy gun may be of particular interest as it has been shown to minimize if not eliminate amorphous damage in GaAs and other materials.
We have several dozen technical papers on ion milling with the IV3 and several papers on the new low energy technology. If you can tell me a little more about your application, I can put together a set of papers th= at would be of interest. Please feel free to contact me off-line if you require any additional information.
NOTE: We do offer an ion mill as described above and therefore do have a vested interest in promoting its use.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by wu jin song } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Hello, everyone,
We used a ion-etching machine without a nitrogen-cooled stage as =
the last step to prepare thin-foil specimens for a long time. =
Now we want to have ion mill that have a cooled stage in order to reduce the formation of large amount of amorphous. Could anybody give us some suggestion on where we can find the ion mill and how much it is? Any information and suggestion about =
it are greatly appreciated.
Jinsong wu LSG2M, Ecole des Mines de Nancy Parc de Saurupt F-54042, Nancy Cedex France
I would suggest using a wire saw with diamond impregnated wire. A diamon= d wheel saw or diamond band saw will also be able to cut it, but will not produce as nice of a surface as the diamond wire saw will. Another optio= n is a high speed diamond saw, but that would typically require embedding t= he sample in a resin of some sort and still will not give you the smooth, polished edge that you will get with a wire saw. The wire saw should giv= e you a cross section suitable for observation without additional polishing= =2E
If you have an interest, please contact me off-line and I will try to put=
you in touch with one of our customers who may be able to do the cutting for you as a service. We may also be able to help you out in our applications lab.
NOTE: We do produce a wire saw, diamond wheel saw and diamond band saw as=
described above and certainly have a vested interest in promoting its use= =2E
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by INTERNET:"corwinl-at-pt.cyanamid.com"-at-sparc5.microscopy.com } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Can anyone suggest how to cut into a glass pharmaceutical vial with =
minimal damage and contamination in order to examine an organic film= =
inside it? Presumably some sort of diamond saw. I need a commercial =
lab who can provide the service, ASAP of course. Hatchet is last =
resort. Thanks for your help. =
=
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400 =
I was wondering as to whether to respond to this question or not. Given the propensity of some to "beat" an issue to death but here goes:
The question as to what to insure an electron microscope (or any piece of equipment) should be replacement value. One can insure anyhting for over or under it's value but the insurance company will only reimburse the insured for the insured value or replacement of the equipment, whichever is less. Believe me, I know this first hand.
In this case one could insure the JEOL JSM-35C for 200K but in case of a total loss the insurace company would not pay the 200K but the replacement value. Next question is what is replacement value and what is the insurance company liable? Are they liable for the $3,800.00 paid for the SEM; the $3,800.00 plus expenses involved in transporting and installing the SEM or another permutation? In my experience, the insurance company would be responsible for the SEM (in a similar condition), installed in the same location. Of course, the insurance company could pay for insured value if it is less.
The bottom line is that in this case (potential insurance claim), labor does add value to equipment. The price paid is not relevant for insurance purposes. I do agree that the resale value does not increase if we take into account transportation and installation costs Supply and demand still prevail. By the way, the prices I quoted for a JEOL 840 includes installation but not transportation. Al Sampson is correct, prices for resale equipment varies as much as 100%, depending upon wholesale or retail values.
Of course someone will continue argue that labor does or does not add value to equipment. To this I say that before an SEM was an SEM it was a "bunch of metal and electronic parts". Labor was needed to make it an SEM and thereby increasing its' value.
As always, I am sure others have a differing opinion. Perhaps we could discuss something relevant like DP oil differences or cleaning RP oil on EDS detectors.
Regards to all,
Earl Weltmer
Marti, Jordi wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Paul: } } I guess I would ask myself how much I could afford if there were to } be major damage to the instruments ( fire, etc) . If replacing the } instruments with my own money represented an "affordable" impact, then } I would not consider putting much money into the insurance. I would } rather spend that money in supplies and or service contracts. } } BTW, I 'm not sure I agree with your assessment that the instruments } will be worth a lot more once they are operating. Of course this will } depend somewhat on the present condition and how much you had to invest } to get them back into operation. } } My thoughts. I am curious to see what others think. } } Jordi Marti } } I have purchased two SEMs, transported them across country, and am } preparing } to set up a lab. Aside from the insurance on the building, I will need } to } insure the equipment inside, hence the question: What is the value of } the } microscopes? } There is of course the purchase price (in my case, $ 3,800 and $ 2,500, } respectively, for a JEOL 35 and Philips 500). Once in place and } running, they } are obviously going to be worth a whole lot more, but how much more???? } I'm } sure my insurance agent will be of no help, as there is no 'blue book' } values } for used SEMs. Any suggestions are most welcome! Thank you. Thank you. } Thank } you. } } Paul Grover } pbgrover-at-aol.com
{HTML} I was wondering as to whether to respond to this question or not. Given the propensity of some to "beat" an issue to death but here goes:
{P} The question as to what to {I} insure {/I} an electron microscope (or any piece of equipment) should be {I} replacement {/I} value. One can insure anyhting for over or under it's value but the insurance company will only reimburse the insured for the insured value or replacement of the equipment, whichever is less. Believe me, I know this first hand.
{P} In this case one could insure the JEOL JSM-35C for 200K but in case of a total loss the insurace company would not pay the 200K but the replacement value. Next question is what is replacement value and what is the insurance company liable? Are they liable for the $3,800.00 paid for the SEM; the $3,800.00 plus expenses involved in transporting and installing the SEM or another permutation? In my experience, the insurance company would be responsible for the SEM (in a similar condition), installed in the same location. Of course, the insurance company could pay for insured value if it is less.
{P} The bottom line is that in this case (potential insurance claim), labor does add value to equipment. The price paid is not relevant for insurance purposes. I do agree that the {I} resale {/I} value does not increase if we take into account transportation and installation costs Supply and demand still prevail. By the way, the prices I quoted for a JEOL 840 includes installation but not transportation. Al Sampson is correct, prices for resale equipment varies as much as 100%, depending upon wholesale or retail values.
{P} Of course someone will continue argue that labor does or does not add value to equipment. To this I say that before an SEM was an SEM it was a "bunch of metal and electronic parts". Labor was needed to make it an SEM and thereby increasing its' value.
{P} As always, I am sure others have a differing opinion. Perhaps we could discuss something relevant like DP oil differences or cleaning RP oil on EDS detectors. {BR}
{P} Regards to all, {BR}
{P} Earl Weltmer {BR} {BR}
{P} Marti, Jordi wrote: {BLOCKQUOTE TYPE=CITE} ------------------------------------------------------------------------ {BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
{P} Paul:
{P} I guess I would ask myself how much I could afford if there were to {BR} be major damage to the instruments ( fire, etc) . If replacing the {BR} instruments with my own money represented an "affordable" impact, then {BR} I would not consider putting much money into the insurance. I would {BR} rather spend that money in supplies and or service contracts.
{P} BTW, I 'm not sure I agree with your assessment that the instruments {BR} will be worth a lot more once they are operating. Of course this will {BR} depend somewhat on the present condition and how much you had to invest {BR} to get them back into operation.
{P} My thoughts. I am curious to see what others think.
{P} Jordi Marti
{P} I have purchased two SEMs, transported them across country, and am {BR} preparing {BR} to set up a lab. Aside from the insurance on the building, I will need {BR} to {BR} insure the equipment inside, hence the question: What is the value of {BR} the {BR} microscopes? {BR} There is of course the purchase price (in my case, $ 3,800 and $ 2,500, {BR} respectively, for a JEOL 35 and Philips 500). Once in place and {BR} running, they {BR} are obviously going to be worth a whole lot more, but how much more???? {BR} I'm {BR} sure my insurance agent will be of no help, as there is no 'blue book' {BR} values {BR} for used SEMs. Any suggestions are most welcome! Thank you. Thank you. {BR} Thank {BR} you.
{P} Paul Grover {BR} pbgrover-at-aol.com {/BLOCKQUOTE} {/HTML}
Prolonged dehydration and embedding will probably do the trick for you. However, I found that the loss of lipid material during a long dehydration protocol is unsatisfactory. Instead I use acidified DMP (Di-methoxy-propane) for chemical dehydration. For small objects, like anthers, complete dehydration can occur in 15 min or less, but I would use one hour at room temp. for a start. DMP can be mixed with all hydrofobic embedding media I know. However DMP dehydration can sometimes reslut in shrinked material as well, but I would give it a try.
Bo
} Dear Rinaldo, my experience with fixation of flowers has been that anthers } are relatively difficult to fix and embed (especially when they are } approching maturity) even when other organs and tissues are well preserved. } If adjusting buffer strength and cutting them transversely as Dr. Echlin } suggests doesn't do the trick, my suggestion would be that you try much } longer dehydration times. I suggest at least an hour at each dehydration } step and perhaps leave them overnight at the 70% ethanol stage. Of course } the trade-off might be that you extract some of the stuff you wish to } preserve. You will have to adjust your protocol on a species to species } basis. John
Dear microscopists, We are planning to buy some diamond knives and I would like to know your impressions of knife performance (what is the difference between Diatome, Drukker, Pelco, DDK etc.). We need standard ultramicrotomy knives for cutting epon embedded biological material and cryo dry ultramicrotomy knives. Personally, I like Diatome and dislike DDK. Did anybody see the difference in cutting and durability between 45=B0 an= d 35=B0 knives?=20 Which boat for standard knife is preferable? What is the purpose of boats with inclined cavity? Any experience is wellcome.
___________________________ Dr. Alexander A. Mironov Jr. Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
Contact Tom Hayes via The Donner Lab, University of California, Berkeley. He has done a lot of work on the SEM of fly ash.
Patrick Echlion Cambridge University
On Thu, 29 Oct 1998, EVERETT RAMER wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are interested in using the SEM to measure the concentration of } flyash in the atmosphere that has come from coal-fired power plants. } One of the issues involved is being able to distinguish particles from } power plants from particles originating at other sources. We would } appreciate any information people might have on the identification of } flyash and determination of its origin. } Everett Ramer } Federal Energy Technology Center } }
Gatan make ion mills with liquid nitrogen cooled stages. We have two Gatan DuoMills here at Birmingham. I don't know how much they cost but they certainly aren't cheap.
Gatan can be reached at http://www.gatan.com/
Another manufacturer of ion mills is Technoorg-Linda. I know less about them.
Hope this helps
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, or: ianmaclaren-at-hotmail.com Birmingham B15 2TT, http://web.bham.ac.uk/I.MacLaren England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Another option would be LUDL Electronics Products. You can contact them at sales-at-ludl.com I have used them frequently in the past with good success. They will be able to help with staging for most any microscope as you will find out. I believe they have a lower end inexpensive stage called the BIO=POINT that is about 5k with additional video auto focus for an additional $500 or so. Contact them and I am sure they can help you out. Take Care and Good Luck!
C.Passione -----Original Message----- } From: Shalvoy, Richard {rbshalvoy-at-corp.olin.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I'm studying microbiology at Laval University in Quebec city, Canada. I would like to know where I can find informations about Normanski interference contrast microscopy.
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meeting point for developers and users working in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
This e-mail was produced by Ernst H.K. Stelzer EMBL, Heidelberg, Germany =========================================== Dr. Frank-Martin Haar European Molecular Biology Laboratory Cell Biology and Biophysics Programme Light Microscopy Group
The 37th Annual Conference of the Microscopy Society of Southern Africa takes place from Tuesday December 1st to Friday December 4th, 1998.
Titles and authors of the extended abstracts of the conference presentations which will be published in Vol 28 of the Proceedings of the Microscopy Society of Southern Africa are available at the following site:
http://www.ru.ac.za/affiliates/emu/mssa.htm
More information on the conference and the Microscopy Society of Southern Africa is available at the following site:
http://www.uct.ac.za/depts/emu/mssa/index.htm
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
It's big, It's beautiful and it could be yours!! Zeiss Ultraphot For Sale w. 4 sources, Hg, Xe, CsI, and W. Objectives negotiable, Auto. camera system not working. Pls. contact Lynne Garone at Polaroid Corp. 781 -386 1446.
There is a post-doctoral opportunity within the Department of Chemical, Biochemical, and Materials Engineering at the Stevens Institute of Technology to develop and apply techniques associated with spatially-resolved electron energy-loss spectroscopy. Application will be made to study the morphology in biological tissue, in synthetic polymers, and in mixtures of these two. Much of this work will be done in collaboration with Unilever Research.
The ideal candidate will have experience in electron optics and electron energy-loss spectroscopy as well as in techniques of ultramicrotomy and cryo-ultramicrotomy. Most critical, however, is a familiarity with eels, digital processing of spectra, and a willingness to work hard and effectively.
The position will open in November, 1998. The appointment will be for one year with a renewal dependent upon performance and availability of funds.
The Stevens Institute of Technology is a small private university concentrating in disciplines of engineering, science, and technology. It is located on the western bank of the Hudson River immediately across from New York City. The Stevens electron-optics laboratory contains a Philips CM20 FEG TEM/STEM, a Philips CM30 SuperTwin TEM, and a Leo 982 FEG SEM, among other instrumentation.
For further information please contact:
Professor Matthew R. Libera Dept. Chemical, Biochemical, and Materials Engineering Stevens Institute of Technology Hoboken, New Jersey 07030 ph: 201-216-5259 fax: 201-216-8306 mlibera-at-attila.stevens-tech.edu
------------------------------------------ Matthew R. Libera Professor Materials Science and Engineering Stevens Institute of Technology Hoboken, New Jersey 07030 ph: 201-216-5259 fax: 201-216-8306 mlibera-at-attila.stevens-tech.edu -------------------------------------------
I am using Image Tool software. When I do measurements by drawing the marker and double clicking to end the measurements the program sometimes crashes. Has this happened to anyone else? Is so how do you correct the problem?
Michael Ingram Rodel, Inc Polishing Lab 451 Bellevue Rd Newark, DE 19713 (302) 366-0500, ext.: 2545
A colleague of mine needs to publish a micrograph taken in 1971 on a Philips 200 TEM; that fine old antique workhorse that listed magnifications in terms of taps. All we know is that this image was photographed at tap 16. Needless to say we no longer have our manual or calibration records from so long ago.
If we could even get a ball park figure for this magnification, it would help; I realize we're talking large ball park here but we can give estimates!
It seems to me that all sawing methods will create lots of debris, something I thought the request wanted to avoid.
Some time ago, I used to make drinking glasses out of bottles. I would cut off the tops with two methods. The first entailed using a piece of nichrome wire wrapped around the bottle. An electrical current was passed through the wire, heating it to red hot, for just a moment. Sometimes, that would nicely cut the top off of the bottle by itself. Some- times, the application of an ice cube around the heated circle was needed to get the crack to form. The second method used a device that I bought, but could be easily built. It used a glass cutter mounted on a rod, and a cone mounted on an arm that could be adjusted up and down the rod. The cone is placed in the mouth of the bottle, the arm is adjusted so the glass cutter is at the level where the cut is desired, and perpen- dicular to the surface of the bottle at that point. Now a line can be scribed around the bottle with the glass cutter. The second part of the kit was a heavier rod with an angle on one end, and an adjustable cone on the straight part. This rod was passed through the mouth of the bottle with the cone adjusted to set the angled end in the bottle even with the scribe line on the outside. By rocking the rod, and tapping the end inside the scribe line, the crack would form along the scribe line and separate the top from the bottom. Some tricks for clean cuts of glass are clean the surface well before doing the scribe, and do not press too hard while making the scribe. Too much pressure causes the scribed line to be chipped up, and a clean break less likely.
There will still be some debris, but it should be minimal amounts of glass chips.
Hope my description is understandable, and it just might help!
I would appreciate any references people might know of containing descriptions of preparation of metallic uranium alloys (approx.90% uranium) for TEM observation.
Does anyone have experience with this? If so, what methods did you use, and what was the outcome?
Has anyone obtained good samples with ion milling (one of our problems involves U-alloy particles in an aluminium matrix)?
Thanks, Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
Hi everyone, I have encountered considerable difficulty, i.e., zero success, in repeating an immunolabelling experiment. I'm working with LR White embedded cotyledon tissue, fixed 12/95, probing with a polyclonal Ab (rabbit), also made in '95. My secondary is commercial protein A gold. I had very specific labelling when the initial experiment was conducted in '95, but now have no specific label. I have repeated the previous successful protocol, but with no luck. My antibodies have been at -20C, having only been thawed once to aliquot them into smaller volumes. Is it possible that there have been changes in the fixed tissue that cause loss of epitopes? I find this hard to believe, but I'd like to hear from others. Could the antibodies have lost their specificity? I have repeated dilution series experiments, but again, all labellings were no more specific than buffer controls. I should say that the antibodies work fine in Westerns, giving us the same banding pattern we've always seen with these Ab, so I suspect the tissue, not the Ab. Any and all comments, criticisms, speculative remarks, SWAGs, etc. would be most welcome. Much hinges on the success of the labelling. Many thanks, Dwight
************************************************************************* Dwight Beebe Prof. Agrege (Associate Prof.) Institut de recherche en biologie vegetale Universite de Montreal 4101 est, rue Sherbrooke Montreal, (Quebec) H1X 2B2 Canada Tel: 514/872-4563 or -4746 (lab) FAX: 514/872-9406
I have a user who wants to embed rubber extracted from plants. The last time she did it she used glut & OsO4 (concentrations I do not know). The insides of the what she thought were solid rubber particles were extracted or at least they looked extracted (so the samples kinda looked like an inner tube). Do any of you bouncing baby scientists have any suggestions for her? She says this stuff is lipid-like and is a short-chain rubber compound, so she figures its pretty liquid.
Send me your suggestions & I'll bounce them off her & see if any stick.
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Dear microscopists, } We are planning to buy some diamond knives and I would like to know your } impressions of knife performance (what is the difference between Diatome, } Drukker, Pelco, DDK etc.). We need standard ultramicrotomy knives for } cutting epon embedded biological material and cryo dry ultramicrotomy } knives. Personally, I like Diatome and dislike DDK. } Did anybody see the difference in cutting and durability between 45=B0 an= d } 35=B0 knives?=20 } Which boat for standard knife is preferable? What is the purpose of boats } with inclined cavity? } Any experience is wellcome. } =20 } ___________________________ } Dr. Alexander A. Mironov Jr. } Unit of Morphology } Dept. of Cell Biology and Oncology } Consorzio Mario Negri Sud } Via Nazionale, S.Maria Imbaro (Ch) } 66030 Italy } =20 } Tel. 0872-570-332 } Fax 0872-578-240 } E-mail: amironov-at-cmns.mnegri.it } =20 } =20 } =20 } =20 I have used DDK, Dupont, Diatome, others. For the last ten years we have invested only in Diatome. For the last seven years we have about 22 thousand dollars worth of assorted Diatome knives. We have Never had a bad one, never had one poorly resharpened (I long ago quit testing them when they came back to us), never had one that wore out quickly. Furthermore, Diatome USA has a laboratory set up. In case you have trouble with your embedding or materials, you can send them some of your tissues with your knife, and they will probe the situation for you and give you advice. I have not used this service, but I know from others that this service is fine. We have gotten excellent advice also on solving problems with our microtomes which was affecting the knives. I don't own stock in Diatome. Wish I did.
I have had a request to examine an interface structure which, while nominally planar, has roughness associated with it. In addition to imaging the structure, the person making the request would like to have a measurement of the roughness associated with the interface. The interface of interest is etched within Si and thus inaccessible to AFM.
Is anybody aware of techniques for measuring interface roughness from a TEM image? Are there any software packages which have such function built-in?
Phil
Philip L. Flaitz IBM Analytical Services Ph.......(914) 892-3094, FAX -2003 flaitz-at-us.ibm.com
In considering your labeling problem, you didn't mention the specifics of the failure. Is your background to high (a potential problem with blocking agents), or do you see no gold at all (a potential problem with the primary and/or secondary or with access of primary to antigen)? How old is your secondary? Our experience suggests that the larger gold conjugates seem to loose their reactivity relatively soon after purchase (even if the packaging suggests that the expiration date is far in the future, they might be optimistic). I have never run across a problem with antibody stored frozen, and routinely use ABs that are at least 10 years old. Is there a particular reason you are using PA/gold? You will get more gold particulates per AB (ie, higher signal) using a GAR conjugated particulates. Your signal will also substantially increase using small (5 nm) particles vs. larger particulates. What is the gold size you are using now? You should consider using the smallest particles you can image at the desired magnification. It might also be that the characteristics of the LR White resin has changed. Do you notice any difference in the cutting properties as compared to earlier use? You might want to try etching the surface of the LR white ever so briefly (though I usually consider this a desperate act). How is the condition of your blocking agents? Serum stored for considerable duration may have failed.
You might have forgotten to utter the magic words.
Good luck,
Doug
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
Lucky for you that I still have my EM records from years ago handy. I got some of my nicest images on that old workhorse. I have the mag table posted to the inside of the frontcover for our old Philips EM200.
Tap 16 was 23,758X for the calibration done when I was using it, and the calibration was done with the projector control set at zero and the voltage at 60kV. An earlier calibration gaqve a mag of 22,400X.
This Philips EM200 used to be at the University of Colorado, Boulder in the Department of Molecular, Cellular and Developmental Biology. It was sold to Colorado State University in 1980, probably to the Department of Anatomy and Neurobiology. It may still be there.
Hope this helps.
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 73-384 CHS 951761 Los Angeles, CA 90095-1761
-----Original Message----- } From: Peggy Brannigan [mailto:brannign-at-asrr.arsusda.gov] Sent: Friday, October 30, 1998 2:23 AM To: Microscopy-at-Sparc5.Microscopy.Com
Hello Listers and history buffs,
A colleague of mine needs to publish a micrograph taken in 1971 on a Philips 200 TEM; that fine old antique workhorse that listed magnifications in terms of taps. All we know is that this image was photographed at tap 16. Needless to say we no longer have our manual or calibration records from so long ago.
If we could even get a ball park figure for this magnification, it would help; I realize we're talking large ball park here but we can give estimates!
I have one reference that I was able to fins quickly for TEM preparation = of Uranium Silicide. The reference is as follows:
"Technique for Preparation of TEM Foils from Two Phase Uranium Silicide",=
B.J. Kestel, Ultramicroscopy 25 (1988) 91-92.
Bernie Kestel is at Argonne National Lab in Illinois and is a great reference for many difficult to thin materials. Almost all of his work i= s done using our Model 550 Single Vertical Jet ElectroPolisher as is the wo= rk done in the referenced paper. We have an extensive bibliography on sampl= e preparation techniques and hope to have that indexed and searchable on ou= r web site by January 1.
I hope this helps!
NOTE: We produce the equipment referenced in this message and have a vest= ed interest in promoting its use.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "Wharton Sinkler" } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Dear List,
I would appreciate any references people might know of containing descriptions of preparation of metallic uranium alloys (approx.90% uraniu= m) for TEM observation. =
Does anyone have experience with this? If so, what methods did you use, and what was the outcome? =
Has anyone obtained good samples with ion milling (one of our problems involves U-alloy particles in an aluminium matrix)? =
Back when God was a boy, Ted Nolan and I did some TEM work on a U-Nb-Zr alloy (Mulberry). I may be able to locate a report on the specimen prep techniques we developed. I recall it was primarily electropolishing, as the work pre-dated the development of ion mills.
Larry
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
I am using Image Tool software. When I do measurements by drawing the marker and double clicking to end the measurement the program sometimes crashes. Has this happened to anyone else? Is so how do you correct the problem? Michael Ingram Rodel, Inc Polishing Lab 451 Bellevue Rd Newark, DE 19713 (302) 366-0500, ext.: 2545
Has anyone ever used tannic acid in their gluteraldehyde? I've heard it's suppose to help preserve cilia and fibers. Any receipes, methods, or concentrations would be appreciated. Thanks in advance.
In a message dated 98-10-30 14:43:30 EST, psic-at-uclink4.berkeley.edu writes:
{ { I have a user who wants to embed rubber extracted from plants. } }
Hi Paula,
Hmm... how novel, bouncing ideas about rubber off of us! I have done some work with thin sectioning of rubber-related products (although synthetic, not natural compounds). I found that cryoultramicrotomy was the best approach.
Is it absolutely necessary for your friend to embed the compounds in plastic? If not, try embedding in sucrose on the head of a cryopin and plunge-freezing in liquid nitrogen, just like you would do for biological material, which is what this stuff is anyway.
I imagine it will take some playing around to find a suitable sectioning temperature, but you should probably start around -60 to -80. Do you have a cryoultramicrotomy setup at Berkeley? If not, try UCSF.
Good luck, hope this helps.
Bob Chiovetti Microimaging Technoogies, Inc. Tucson, Arizona USA Tel./Fax (520) 546-4986 rchiovetti-at-aol.com
We just acquired a new SEM in our diagnostic pathology lab, located in New York. It is a partial pressure scope, from Philips. Has anyone here done any work with SEM and diagnostic pathology? Have any ideas you would like to discuss, or projects?
Dear microscopists, Does anybody know e-mail or fax of Agar Scientific in US? Thank you in advance.
___________________________ Dr. Alexander A. Mironov Jr. Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
Hello Alexander, I don't know the US details but the Agar Scientific head office in the UK will, their fax number is (+44) 1279 815106. Good luck,
Ron
On Sun, 1 Nov 1998, Alexander Mironov Jr. wrote: } } } Dear microscopists, } Does anybody know e-mail or fax of Agar Scientific in US? } Thank you in advance. } } ___________________________ } Dr. Alexander A. Mironov Jr. } Unit of Morphology } Dept. of Cell Biology and Oncology } Consorzio Mario Negri Sud } Via Nazionale, S.Maria Imbaro (Ch) } 66030 Italy } } Tel. 0872-570-332 } Fax 0872-578-240 } E-mail: amironov-at-cmns.mnegri.it } } } }
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Eastman Chemical Company has an opening for a Technician in the Physical Chemistry Research Laboratory. A candidate should have a 2-year degree, or higher, and experience in optical and/or electron microscopy. A background in chemistry is preferred but not required. The duties will include the use of optical and electron microscopes in support of research projects and in solving problems relating to products and manufacturing. The laboratory is fully state-of-the-art and offers varied and challenging assignments.
Eastman Chemical Company is one of the largest chemical, fiber and plastics manufacturing facilities in the U.S. The company's central research laboratories are in Kingsport, Tennessee, and provide a variety of resources for the technical community. Kingsport is located in northeastern Tennessee in the foothills of the Smoky Mountains. The area is rated among the top 25 most livable metropolitan areas and offers affordable housing, low taxes, and excellent schools. The pleasant four-season climate permits a variety of outdoor recreational opportunities.
Interested persons should send their resume to: Eastman Chemical Company, Employment Department, P.O. Box 1975, Kingsport, TN USA 37662-5215. Eastman Chemical Company is an Equal Opportunity Employer.
Dennis B. Barr (dennbarr-at-eastman.com) Physical Chemistry Research Laboratory Physical & Analytical Chemistry Research Division Eastman Chemical Company Kingsport, TN 37662-5150
In reference to embedding rubber from plants. So the rubber has been extracted from the plant and she wants to look at the suspension of rubber particles. Natural rubber or latex derived from plants is cis-polyisoprene -(CH2-C2H3=CH-CH2)- , which has an available double bond. If she has a suspension, I wouldn't extract, but just go ahead and dilute the latex, and then stain with a saturated bromine solution, or osmium tetroxide. A droplet can be placed on a carbon film on a grid. If cross-sections are necessary, cryo (as mentioned in an earlier E-mail) is an option. Or maybe she could dry a film, and harden the material with osmium prior to RT sectioning. Just thoughts on a Monday morning. Take care, Vicky
Does anybody know a supplier of the old-style scotch tape that can be used to make thin TEM samples? It can be used to remove thin layers of samples. You could easily make a nice graphite sample this way. I appreciate any help I can get. Thank you.
Peggy Bisher.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
Have you tried a different batch of protien a-gold? I've only done this kind of experiment once, a long time ago, but when I had problems this was the problem.
Karen Pawlowski
On Fri, 30 Oct 1998, Dwight Beebe wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi everyone, } I have encountered considerable difficulty, i.e., zero success, in } repeating an immunolabelling experiment. I'm working with LR White } embedded cotyledon tissue, fixed 12/95, probing with a polyclonal Ab } (rabbit), also made in '95. My secondary is commercial protein A gold. I } had very specific labelling when the initial experiment was conducted in } '95, but now have no specific label. I have repeated the previous } successful protocol, but with no luck. My antibodies have been at -20C, } having only been thawed once to aliquot them into smaller volumes. } Is it possible that there have been changes in the fixed tissue } that cause loss of epitopes? I find this hard to believe, but I'd like to } hear from others. Could the antibodies have lost their specificity? I } have repeated dilution series experiments, but again, all labellings were } no more specific than buffer controls. I should say that the antibodies } work fine in Westerns, giving us the same banding pattern we've always seen } with these Ab, so I suspect the tissue, not the Ab. } Any and all comments, criticisms, speculative remarks, SWAGs, etc. } would be most welcome. Much hinges on the success of the labelling. } Many thanks, } Dwight } } } ************************************************************************* } Dwight Beebe } Prof. Agrege (Associate Prof.) } Institut de recherche en biologie vegetale } Universite de Montreal } 4101 est, rue Sherbrooke } Montreal, (Quebec) H1X 2B2 Canada } Tel: 514/872-4563 or -4746 (lab) } FAX: 514/872-9406 } } } }
When this kind of thing happens to me, I recheck the antibody on fresh frozen or what ever Light Level tissue sections you know it should work on. If the staining pattern is the same as the original experiment, then it would lead you to the tissue. I have seen LRWhite tissue sections become unusable in matter of days or weeks sometimes, but the blocks have been pretty stable. I have also seen some strange staining pattern when the pH of the buffer is off.
Good luck!
Derm Imaging Center University of Washington
On Fri, 30 Oct 1998, Dwight Beebe wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi everyone, } I have encountered considerable difficulty, i.e., zero success, in } repeating an immunolabelling experiment. I'm working with LR White } embedded cotyledon tissue, fixed 12/95, probing with a polyclonal Ab } (rabbit), also made in '95. My secondary is commercial protein A gold. I } had very specific labelling when the initial experiment was conducted in } '95, but now have no specific label. I have repeated the previous } successful protocol, but with no luck. My antibodies have been at -20C, } having only been thawed once to aliquot them into smaller volumes. } Is it possible that there have been changes in the fixed tissue } that cause loss of epitopes? I find this hard to believe, but I'd like to } hear from others. Could the antibodies have lost their specificity? I } have repeated dilution series experiments, but again, all labellings were } no more specific than buffer controls. I should say that the antibodies } work fine in Westerns, giving us the same banding pattern we've always seen } with these Ab, so I suspect the tissue, not the Ab. } Any and all comments, criticisms, speculative remarks, SWAGs, etc. } would be most welcome. Much hinges on the success of the labelling. } Many thanks, } Dwight } } } ************************************************************************* } Dwight Beebe } Prof. Agrege (Associate Prof.) } Institut de recherche en biologie vegetale } Universite de Montreal } 4101 est, rue Sherbrooke } Montreal, (Quebec) H1X 2B2 Canada } Tel: 514/872-4563 or -4746 (lab) } FAX: 514/872-9406 } } } }
Interface roughness in GaAs/AlGaAs was a hotly debated subject a few years back. Optical techniques seemed to indicate a perfect, atomically flat interface with occasional steps, HRTEM showed roughness at the atomic scale. I was involved with the HRTEM part. We used a technique developed by Ourmazd et. al. at Bell Labs to analyze the HRTEM cross sections. We called it "chemical mapping" or "chemical lattice imaging". I don't know if this helps, as you don't say, what exactly your interface consists of, but try a literature search for the above keywords. Also try to search for "Ourmazd". Other authors on various publicatons were Kim, Baumann, Warwick, and myself. I have been out of the field for a few years, but Ourmazd and coworkers have extended the analysis to SiGe systems as well.
The software we used was developed by a number of people at Bell Labs and ran on a Silicon Graphics. At the time we were giving away the software to other researchers. I don't know, if that is still the case. The group at Bell Labs does not exist anymore, Ourmazd is now in Frankfurt/Oder in Germany.
Good luck.
Michael Bode Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS fax: (303) 234-9271 email: info-at-soft-imaging.com
} } } ---------- } From: Philip Flaitz[SMTP:flaitz-at-us.ibm.com] } Sent: Friday, October 30, 1998 1:05 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM: interface roughness analysis. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is a summary of the replies I rec'd to my inquiry about beam damage to carbon tape. Thanks to all who replied.
Owen
+++++++
My initial message was:
Good morning:
We jet spray volcanic ash particles (~10 microns or less) onto carbon double stick tabs for SEM/EDS study. It is easy to produce a nice distribution of particles with few that overlay one another. Problem is that later, during EDS and backscatter imaging, when the beam current is higher, the carbon tape is often damaged. It curls and bubbles thereby distorting the images.
We tried to use carbon paint but it is tricky to get the right amount spread onto the mount or it dries too quickly. Results were unfavorable.
I'd really like to have a durable, very smooth, low Z surface, that is insensitive to beam current and is easy to apply and work with in the SEM. Oh, it should also be economical too.
Any ideas? TIA
Owen
+++++++++
The carbon tape is successful with distributing the effects of charging but not heat. I, at least, can only suggest you sputter with a heat conducting metal ... e.g., Al, Au, Cu ... most any metal would help, but I realize it would superimpose spectra and degrade your BSE contrast. Short of that, you'd need to lower the energy in your beam with either lower beam currents or accel voltage. I'll also be looking for other ideas ... hope this helps :o)
cheerios, shAf Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
+++++++++
I have never had damage to carbon tape, even at 30kV. I store mine in the fridge when not in use, as I heard it deteriorates. I bought my rolls from SPI several years ago and never had a problem.
Alan Stone ASTON Metallurgical Services
+++++++++
I would like to suggest Berylium as a possible solution for a smooth, low-Z surface. It can be evaporated onto the specimen and it satisfies all the given criteria. HOWEVER, there are very serious safety issues involved. If you have access to the appropriate equipment--a dedicated vacuum evapora- tor with exhaust vented appropriately, the equipment to produce small Be pellets or wire for evaporation, a safe place to clean out the bell jar, etc.--you might want to consider using Be. I think that solid Be metal is not too dangerous, and that finely-divided metal or compounds are the most hazardous. I have handled BeO powder (using a hood, of course) and had no trouble, and I heard a presentation at MSA some years ago by Dr. Hall (whose first name escapes me) where Be coating was used instead of C for low temper- ature work (where Be's conductivity is much greater than C's). With the stated safety reservations, perhaps you could produce a Be equivalent of double-stick tabs. Good luck. Yours, Bill Tivol
++++++
Owen,
Have you tried double sided conductive carbon sheets, instead of the tabs? While the systems appear to be similar, the sheet product is a bit more resistant to changing in the electron beam. Now if that either fails as well, or is not something you would want to try, there is a method I have described on the listserver at least once before and maybe twice. It is based on the use of a camphor naphthene eutectic composition. For the coverglass smoothnes, purchase HOPG materials. would seem like for your appliction, being non-SPI, that the cheapest grade would be acceptable for you. Remember that is just some judgement on my part but it might not be correct. But you want to make a freshly cleaved surface to expose the nice smooth surface of HOPG. I beam won't hurt it in the least. I guarantee that. Then use the camphor naphthalene procedure to disperse the particles on the HOPG. A quick and dirty verion of this might be to put the HOPG on a hot plate at something around 100 deg. C, and disperse your ash particles in acetone, the acetone will evaporate away before the particles have time to agglomerate. But if they don't, then you should turn to the camphor naphthalene method.
Chuck
++++++++
Ihave quite often used conductive carbon tape for such exams with no problem.
.Wonder what beam current and voltage you are running? I presume you are carbon coating the ash/tape to aviod charging and that is not the distortion
problem.
Woody White McDermott Technology, Inc. +++++++
Owen, Since you don't need much adhesive to tack down 10 micron particles, you may try applying a very thin layer of a fluid adhesive (such as Microstick) to a pyrolytic carbon planchet. This form of carbon has a flat, hard, glass-like surface. A drop of adhesive applied to the carbon may be spread very thin by drawing out with a cover slip. Since the organic adhesive layer is thin, you will probably get your required conductivity. There is no observable background structure. Planchets are about $35 each.
Dennis.
Dennis C. Ward voice: 202-324-2982 FBI fax: 202-324-4018 Microanalysis Laboratory e-mail: DCWard-at-concentric.net
+++++++++
The only way to do this is with Carbon Dag , what you do is take a match dip it into the dag and with the match level to the stub just pull across stub only pull match across once this ensure quite a flat level. Just be sure to dilute the dag quite a bit this gives you more of a flat base.
Luc Harmsen Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
++++++++
As far as the tape goes, we use a 3M double back tape that seems to work fairly well so long as you carbon coat the samples. You may want to give it a try. We have also developed an ethyl acetate copolymer that we apply to mylar film which seems to work even better.
Keith Rickabaugh Manager, Materials and Particle Characterization {krickabaugh-at-rjlg.com}
RJ Lee Group, Inc. 350 Hochberg Road Pittsburgh, PA 15146 ph: 724-325-1776 {www.rjlg.com}
+++++++++
Dennis has a good idea. I would substitute freshly-cleaved mica for the carbon planchet...its a lot cheaper.
Chuck Butterick Engineered Carbons, Inc .
+++++++++
There is a product that might fill your bill. It is a thermoplastic called Tempfix marketed by Electron Microscopy Sciences (EMS). In '94 we published a paper in Trans. Am. Microsc. Soc."Mounting Pollen on a Thermoplastic Adhesive for Scanning Electron Microscopy" in which we describe our protocol and demonstrate its advantages for our use. While we did not do any x-ray or BSE imaging on those samples, I believe it would hold up to such use (depending ofcourse on the accel. voltage and beam current used). If you cannot find the journal, I would be glad to send you a reprint if you're interested. Please contact me off line. I have no interest in EMS except as a customer.
Bill
+++++++++
Remember also that if you are not concerned about EDS peaks from the substrate background, consider Tacky Dot Slides. Those are really the ideal kind of mounting system if you are trying to catalog what you have in some quantitative basis, and they also make it easy to come back to the same particle characterized previously.
Chuck
+++++++
Owen
There IS an adhesive that is: smooth-surfaced, high tack, low Z, durable, easy to use, dependable, nontoxic and inexpensive. It is also pretty stable under the beam and thin layers do not outgas noticeably in the vacuum. It is, however, not conductive (after all - is there such a thing as a perfect whatever?). We use this stuff regularly to stick down pollen, fly-ash, small insects, blood cells, ceramic powders, pigment particles etc. for imaging and for EDS analysis.
it is called 'Artists Gold Size' and is a thick tacky syrup of partially polymerised liseed oil. On exposure to air it polymerizes and forms an insoluble polymer (this was used in the manufacture of old-style floor linoleum, hence the name linoleum - from 'linseed oleum').
Buy this at any art supply shop, spread a very small drop of the syrup over the surface of a stub, wait until it is just tacky enough not to wick up the sample, drop your volcanic ash particles onto the surface and leave for 30 min. at room temp (or a few min at 50C) to polymerize, coat with carbon and analyse.
I've never been able to find out who used this first, but have been using it for many years after coming across a casual reference to its use as a SEM mountant.
A $5 bottle should last you years.
Pity it isn't conductive though.
Jan C
============================= Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
Tannic acid is usually used with both the primary and the post fixation in 1-8% (w/v) concentration. The protocol I used successfully before was as the follows.
The fresh made solution of 12% tannic acid (Polysciences cat#4459, EM grade) buffered in 0.2 M cacodylate, pH 7.4 was mixed in an equal volume ratio immediately before fixation with 5% glutaraldehyde in 0.2 M cacodylate buffer, pH 7.4. The final pH is adujsted to 7.2 with 10 N NaOH. The tissues are then postfixed with 2% OsO4 only in the same buffer or a mixture of tannic acid with OsO4.
Good luck,
Ming
} Has anyone ever used tannic acid in their gluteraldehyde? I've heard it's } suppose to help preserve cilia and fibers. Any receipes, methods, or } concentrations would be appreciated. Thanks in advance.
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * #1074B Dentistry Pharmacy Building * * University Of Alberta. * * Edmonton, Alberta, Canada T6G 2N8 * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
This is one of those challenging imaging problems. There are several companies, including Wyko (now Veeco) and Zygo, which specialize in surface structure analysis. Both do 3-D interferometry. I am not sure that they are able to see throughh one layer into another and measure the roughness of the second surface, but it is worth a try. Both have web sites. If you can't get info, contact me off-line and I will supply contact names and numbers.
Dear Mike, When you do measurements in Image Tool, you should just hold down the mouse at the first point and release it at the second. The double-click is confusing the software. I am using the latest version (1.28) on the site. You wrote: } } I am using Image Tool software. When I do measurements by drawing the } marker and double clicking to end the measurement the program sometimes } crashes. Has this happened to anyone else? Is so how do you correct } the problem? } Michael Ingram } Rodel, Inc
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Hi I have neural tissue embedded in Unicryl by way of UVlight at 4 degrees C. It was fixed with 4% paraform and 0.5% glut, no osmium. I am having difficulty staining with 2% aquas Ua and Renolds lead citrate. I have tried combinations from 3 min to as much as 10 min with very little luck, yet the same stains work with my standard 812 embedded sections at 6 min and 5 min respectively. My past experience with LRW was that it took less time and or concentrations than epon like plastics. The T-Blue stained very quickly like LRW. 1. Anyone out there have some experience or ideas? 2. What kind of time periods should one use for washes in drops of distilled water after Ua?
We would like to attach a small hydrophobic protein to small gold particles (less than 5nm). The problem is that most recipes are for proteins in aqueous solutions. We would like to put the gold into chloroform/methanol since our protein is happiest in that, and have the labelling proceed. Does anyone have experience with this?
We have tried mixing protein and gold each in 70% isopropanol since both are happy in that; but we get precipitate when we dry the mixture down.
Gold can be prepared (and obtained from Nanoprobes) with attached alkanes to which our protein might be adsorbed but we would like to avoid increasing the size of the particles by this intermediate layer and further wonder if such an extra layer might not weaken the protein-gold linkage, the protein come off when added to the tissue and gold stick non-specifically to other things.
We can't do immunogold since the protein is likely buried in lipid bilayers and has not been possible to label with conventional aqueous antibody-gold. The protein is surfactant protein B (Possmayer, Voorhout, Hawgood).
Since the gold-protein bond is said to be hydrophobic (and the degree of hydrophobicity of a protein to correlate with its stickability) we hope that the gold should stick to the protein.
Any thoughts will be appreciated, Jacob
Jacob Bastacky, M.D. Room 116 Donner Laboratory Lawrence Berkeley Laboratory University of California Berkeley, California 94720 Telephones: 510.486.4606 office, 510.486.4605 lab, 510.845.8031 fax email: sjbastacky-at-lbl.gov
6 weeks ago I posted an inquiry about color slide photography of computer screens. Lots of helpful replies made my little project an easy success.
I used Kodak EliteChrome ASA 100, f5.6 to 8, 0.5 second exposure and midrange brightness and contrast on the monitor. A tripod and cable release were employed. The camera was placed about 3 feet from the screen and a 58mm lens was used.
The resulting slides showed faithful color and were sharp enough to pass for original slides to the unsuspecting.
The main difficulties were in framing, and in distortions relating centering and perpendicularilty of the camera.
A black mat cut to the aspect ratio of the camera was used to solve the framing problem.
The distortions were more troublesome and not entirely overcome. At three feet slight off axis camera placement creates noticeable taper. A carefully made perpendicular "hood" mount would help. Bill Miller suggested mounting a small mirror on the center of the screen and looking for a reflection right back down the lens as a means of orienting the camera. I didn't try this. Moving farther away and using a telephoto lens is another suggestion which I didn't try, but should have.
Email: TKGreen-at-aol.com Name: Tristan Green School: Piedmont High School
Question: Do you know of a good place to find pictures of common bacterial cultures to use for identification purposes? We are culturing colonies on petri dishes from swabs taken around campus. I would love for my students to be able to identify what they've found.
Thank you, Mrs. Tristan Green Science Teacher, Piedmont High School
6 weeks ago I posted an inquiry about color slide photography of computer screens. Lots of helpful replies made my little project an easy success.
I used Kodak EliteChrome ASA 100, f5.6 to 8, 0.5 second exposure and midrange brightness and contrast on the monitor. A tripod and cable release were employed. The camera was placed about 3 feet from the screen and a 58mm lens was used.
The resulting slides showed faithful color and were sharp enough to pass for original slides to the unsuspecting.
The main difficulties were in framing, and in distortions relating centering and perpendicularilty of the camera.
A black mat cut to the aspect ratio of the camera was used to solve the framing problem.
The distortions were more troublesome and not entirely overcome. At three feet slight off axis camera placement creates noticeable taper. A carefully made perpendicular "hood" mount would help. Bill Miller suggested mounting a small mirror on the center of the screen and looking for a reflection right back down the lens as a means of orienting the camera. I didn't try this. Moving farther away and using a telephoto lens is another suggestion which I didn't try, but should have.
The TEM group of Abbas Ourmazd has moved to the Institute for Semiconductor Physics in Frankfurt (Oder), Germany. The group continues to apply quantitative HRTEM and electron holography. Peter Schwander is in charge for maintaining the Chemical Mapping and QUANTITEM software. This software is ready to run on SGI workstations under IRIX 5.3 and 6.2.
Anyone interested in Chemical Mapping or QUANTITEM or any related information is welcome to contact Peter Schwander ( schwander-at-ihp-ffo.de ).
Alex *=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*= =B0* Dr. Alexander Orchowski Institute for Semiconductor Physics Frankfurt (Oder), Germany http://www.ihp-ffo.de phone: +49 (0) 335 562 5432 fax: +49 (0) 335 562 5300 email: orchowski-at-ihp-ffo.de *=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*=B0*= =B0*
Michael Bode wrote:
} Phil,
} Interface roughness in GaAs/AlGaAs was a hotly debated subject a few } years back. Optical techniques seemed to indicate a perfect, atomically } flat interface with occasional steps, HRTEM showed roughness at the } atomic scale. I was involved with the HRTEM part. We used a technique } developed by Ourmazd et. al. at Bell Labs to analyze the HRTEM cross } sections. We called it "chemical mapping" or "chemical lattice imaging". } I don't know if this helps, as you don't say, what exactly your } interface consists of, but try a literature search for the above } keywords. Also try to search for "Ourmazd". Other authors on various } publicatons were Kim, Baumann, Warwick, and myself. I have been out of } the field for a few years, but Ourmazd and coworkers have extended the } analysis to SiGe systems as well.
} The software we used was developed by a number of people at Bell Labs } and ran on a Silicon Graphics. At the time we were giving away the } software to other researchers. I don't know, if that is still the case. } The group at Bell Labs does not exist anymore, Ourmazd is now in } Frankfurt/Oder in Germany.=20
} Good luck.
} Michael Bode } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } phone: (888) FIND SIS } fax: (303) 234-9271 } email: info-at-soft-imaging.com =20 } Philip L. Flaitz wrote:
} } I have had a request to examine an interface structure which, while nominally } } planar, has roughness associated with it. In addition to imaging the } } structure, the person making the request would like to have a measurement= of } } the roughness associated with the interface. The interface of interest is } } etched within Si and thus inaccessible to AFM.
} } Is anybody aware of techniques for measuring interface roughness from a= TEM } } image? Are there any software packages which have such function built-in?
} } Phil
} } Philip L. Flaitz } } IBM Analytical Services } } Ph.......(914) 892-3094, FAX -2003 } } flaitz-at-us.ibm.com
Something we and others have experimented with is the use of Dimethylsulfoxide (DMSO). It acts as a cell penetrant and stabilizes the cytoskeleton. My work is principally with human neutrophils but others have used it for yeast cells (and other things, but this should be enough to test the idea.)
References:
Gilbert, CS and RT Parmley Morphology of human neutrophils: A comparison of cyrofixation, routine glutaraldehyde fixation, and the effects of dimethyl sulfoxide Anat Rec 252:254-263 (1998)
Fassel, TA; Sohnle, PG and VM Kushnaryov The use of dimethylsulfoxide for fixation of yeasts for electron microscopy Biotechnic & Histochemistry 72(5):268-272 (1997)
Forgive my Portuguese, it's little rusty. I can help with acquiring copies of the references if you need them.
Ciao,
Chuck
--- On Wed, 28 Oct 1998 20:37:25 -0200 Rinaldo Pires dos Santos {rinaldop-at-botanica.ufrgs.br} wrote: ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear colleagues
I am working with the ultrastruture of anthers of Ilex paraguariensis. However, during the dehydration of the samples, fixed in a mixture of glutaraldehyde 2,5% and formaldehyde 2%, the anthers shrink, mainly after the secondary fixation with osmium tetroxide. I used acetone or ethanol , in steps of 30, 50, 70, 90, 90, 100, 100, with 15 minutes in each step, at room temperature. The anthers has a very reduced dimension (about 1 mm length) and shrink in the step 100. What to do? Should I to use a low temperature (4oC)? Thanks in advance.
-----------------End of Original Message-----------------
------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
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On Fri, 30 Oct 1998, Alexander Mironov Jr. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Dear microscopists, } We are planning to buy some diamond knives and I would like to know your } impressions of knife performance (what is the difference between Diatome, } Drukker, Pelco, DDK etc.). We need standard ultramicrotomy knives for } cutting epon embedded biological material and cryo dry ultramicrotomy } knives. Personally, I like Diatome and dislike DDK. } Did anybody see the difference in cutting and durability between 45=B0 an= d } 35=B0 knives?=20 } Which boat for standard knife is preferable? What is the purpose of boats } with inclined cavity? } Any experience is wellcome. } =20 } ___________________________ } Dr. Alexander A. Mironov Jr. } Unit of Morphology } Dept. of Cell Biology and Oncology } Consorzio Mario Negri Sud } Via Nazionale, S.Maria Imbaro (Ch) } 66030 Italy } =20 } Tel. 0872-570-332 } Fax 0872-578-240 } E-mail: amironov-at-cmns.mnegri.it } =20 } =20 } =20 } =20 I have used DDK, Dupont, Diatome, others. For the last ten years we have invested only in Diatome. For the last seven years we have about 22 thousand dollars worth of assorted Diatome knives. We have Never had a bad one, never had one poorly resharpened (I long ago quit testing them when they came back to us), never had one that wore out quickly. Furthermore, Diatome USA has a laboratory set up. In case you have trouble with your embedding or materials, you can send them some of your tissues with your knife, and they will probe the situation for you and give you advice. I have not used this service, but I know from others that this service is fine. We have gotten excellent advice also on solving problems with our microtomes which was affecting the knives. I don't own stock in Diatome. Wish I did.
Since the release of Time magazine's Oct. 12th article concerning the Deep Space I project, which is now heading for an asteriod orbiting the sun, we have been inundated with inquiries concerning our electron microscope apertures. Let me try to answer some of the questions posed to us. The xenon ion propulsion system (XIPS) has launched a new era in satellite propulsion technology. Currently three satellites are in geo-synchonous orbit above the earth using XIPS. In conjunction with the satellite designers we have been able to adapt the standard electron microscope aperure in the thruster control devices in the XIPS, which Time magazine described as the " stuff of technological fantasies ". In addition to EM aperures with non-standard holes, ultrathin and special metals, EN type apertures are also now being used in other applications, including:
- FIBS - X-ray collimators and focusing devices - Light and gas control devices requiring precise microholes and slits - Solder droplet production
We believe the advances we have made in these other applications have allowed us to improve EM apertures overall. I hope this answers most of your questions. We are sending out more detailed information to those who requested it.
Thank you for your interest,
John Arnott Chairman --
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
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On Fri, 30 Oct 1998, Alexander Mironov Jr. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Dear microscopists, } We are planning to buy some diamond knives and I would like to know your } impressions of knife performance (what is the difference between Diatome, } Drukker, Pelco, DDK etc.). We need standard ultramicrotomy knives for } cutting epon embedded biological material and cryo dry ultramicrotomy } knives. Personally, I like Diatome and dislike DDK. } Did anybody see the difference in cutting and durability between 45=B0 an= d } 35=B0 knives?=20 } Which boat for standard knife is preferable? What is the purpose of boats } with inclined cavity? } Any experience is wellcome. } =20 } ___________________________ } Dr. Alexander A. Mironov Jr. } Unit of Morphology } Dept. of Cell Biology and Oncology } Consorzio Mario Negri Sud } Via Nazionale, S.Maria Imbaro (Ch) } 66030 Italy } =20 } Tel. 0872-570-332 } Fax 0872-578-240 } E-mail: amironov-at-cmns.mnegri.it } =20 } =20 } =20 } =20 I have used DDK, Dupont, Diatome, others. For the last ten years we have invested only in Diatome. For the last seven years we have about 22 thousand dollars worth of assorted Diatome knives. We have Never had a bad one, never had one poorly resharpened (I long ago quit testing them when they came back to us), never had one that wore out quickly. Furthermore, Diatome USA has a laboratory set up. In case you have trouble with your embedding or materials, you can send them some of your tissues with your knife, and they will probe the situation for you and give you advice. I have not used this service, but I know from others that this service is fine. We have gotten excellent advice also on solving problems with our microtomes which was affecting the knives. I don't own stock in Diatome. Wish I did.
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Dear Mr. Corwin,
1) Encapsulate the sample fully in Epoxy 2) Using a high speed diamond saw, cut the sample with a RESIN Bonded diamond blade to minimize chipping of the edge. 3) If the cut surface is not good enough, a quick polish maybe necessary.
Try HiRel Laboratory in Spokane, Washington, or Metals Technology in Northridge, Ca. Either of them can be found in directory assistance.
Good Luck,
Gary Liechty Allied High Tech Products, Inc. Products for Metallographic, SEM and TEM Sample Preparation 800-675-1118
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Can anyone suggest how to cut into a glass pharmaceutical vial with } minimal damage and contamination in order to examine an organic film } inside it? Presumably some sort of diamond saw. I need a commercial } lab who can provide the service, ASAP of course. Hatchet is last } resort. Thanks for your help. } } } Leonard Corwin } Research Chemist } Fort Dodge Animal Health } Princeton, NJ 08543-0400
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I am attempting to help a colleague who is in a bind. She needs to get QUICKLY - which is to say - borrow, loan, rent, or buy - a computer controlled stage for an Olympus BH-2 optical microscope.
We're talking to Olympus of course but still ...
Anyone have ideas or a stage available???
Richard Shalvoy rbshalvoy-at-corp.olin.com 203-271-4272
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Hey There Listers,
I have a user who wants to embed rubber extracted from plants. The last time she did it she used glut & OsO4 (concentrations I do not know). The insides of the what she thought were solid rubber particles were extracted or at least they looked extracted (so the samples kinda looked like an inner tube). Do any of you bouncing baby scientists have any suggestions for her? She says this stuff is lipid-like and is a short-chain rubber compound, so she figures its pretty liquid.
Send me your suggestions & I'll bounce them off her & see if any stick.
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
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We are interested in using the SEM to measure the concentration of flyash in the atmosphere that has come from coal-fired power plants. One of the issues involved is being able to distinguish particles from power plants from particles originating at other sources. We would appreciate any information people might have on the identification of flyash and determination of its origin. Everett Ramer Federal Energy Technology Center
Has anyone ever tried any Butvar other than 98? I found a jar of Butvar-79 (from Monsanto) in our chemical cabinet and was thinking of giving it a whirl. I'm guessing it is a shorter polymer than the 98. Nobody knows what it is doing here...it seems to have been inherited from the previous inhabitants.
By information about SEM in seeds of Avena sativa and Avena byzantina (oats). Thanks very much
Diana Reinoso
cultivar-at-fca.uner.edu.ar Facultad de Ciencias Agropecuarias Universidad Nacional de Entre R=EDos Rep=FAblica Argentina --------------------------------- Catedra Botanica Sistematica Fac. Cs. Agropecuarias UNER C.C. 24 3100 - Parana - Entre Rios ARGENTINA Tel: 043 - 975075 Fax: 043 - 975096 E-MAIL: botanica-at-fca.uner.edu.ar
Here I am once again, hat in hand to ask for your help. This time I have someone who wants to do SEM on a cell line grown in suspension (they are completely non-adherent, so we can't grow them that way). What should we do in terms of fixation, etc. and then how do we stick them to the stub? Do we try to make them stick to something first & then run them up? Do you run them up first & then stick them to something? Do you run up to them & say "Stick 'em up"? As you can tell, this is a new one to me. I've process little things by wrapping them in lens tissue & then running the up & then shaking the sample onto the sticky carbon dots. I don't think that procedure will work for these because these cells are much tinier. I thank you in advance for any suggestions you can give me. And I thank you for all the help and advice you've given me in the past.
I'll stick by my computer until I hear from y'all,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
1 a supplier of cheap glass slides about 26 x 46 mm, 1 to 1.5mm thick?
2 someone who can take a given powder/dust sample and perform a respirable/non-respirable binary split of it and return to me the two size fractions for subsequent characterisation?
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Postdoctoral Position in Interfacial Segregation Department of Chemical and Materials Engineering University of Kentucky Lexington, KY USA
A postdoctoral position is available in the area of interfacial segregation. The research project is focused on understanding interfacial segregation at ceramic/ceramic and metal/ceramic interfaces and it effects on physical properties of interfaces. Through a collaboration with Prof. Susan Sinnott, a computational materials scientist at the University of Kentucky, the issue of interfacial segregation will be addressed from a combined experimental and theoretical approach. The ideal candidate for this position will have experience in HREM, STEM , EDS and/or EELS. The University of Kentucky is in the process of developing a state-of-the-art microscopy facility and will be purchasing a 200kV Field Emission TEM/STEM in 1999. The University has an active and growing materials science program with many interdiscplinary research and educational activities between the Materials Engineering, Physics and Chemistry Departments and the Center for Applied Energy Research. Many opportunities for performing experiments at Oak Ridge National Laboratory are also available. Please send applications to Professor Elizabeth Dickey at the address below.
***************** Professor Elizabeth Dickey Department of Chemical and Materials Engineering 177 Anderson Hall University of Kentucky Lexington, KY 40506-0046 USA ph: 606.257.8572 FAX: 606.323.1929 email: ecdickey-at-engr.uky.edu
We can't leave you dangling like this. There are several approaches to solve this suspensful situation. I base this on work I have done with bacteria, fungi and tissue cultures grown in suspension.
In the first approach (easy but yielding few, isolated cells) , take a scrupulously clean, glass microscope slide (use a good glassware detergent and rinse it quite well - do not use acid cleaning agents as they may be toxic). Treat the glide with a poly-L-lysine solution (1 mg/ml distilled water) for 5 minutes, then rinse with distilled water. Place slide into a petri dish containing a filter paper moistened with distilled water and containing several pieces of applicator stick. The slide should be suspended (like a bridge) over the applicator sticks. Gently layer on the cell suspension on the slide so that it fills the slide but does not overflow the edges (about 5 ml). Allow the cells to settle onto the slide for about 1 hr. You should keep the cells "happy" during this time period (out of the light, warm, perhaps in a CO2 incubator). Now, very carefully aspirate away most of the liquid and very slowly add 5 ml of your primary fixative (4% formaldehyde/2%glutaraldehyde) to again cover the slide. Allow to fix for 15-20 min at room temp. Rinse by gently decanting and gently adding buffer from one end of the slide. Post-fix in 2% osmium tetroxide (buffered or in distilled water) for 30 min at room temp. Rinse in distilled water and dehydrate the entire slide by overlaying with 50, 75, 95, ABS ethanol. They key here is "gently". After absolute ethanol, critical point dry the slide (or portion of the slide), coat with conductive metal and examine in the SEM. This technique will give you beautiful black backgrounds with only a few cells per field of view. Some people prefer this method since the cells are quite isolated and not piled up.
A second approach (possibly yielding too many cells on a substrate), is to pass the cell suspension through a micropore filter or silver membrane (available through Structure Probe, for example) so that the cells are retained on the surface of the membrane. This must be done very gently so that the cells are not broken. We have best success by sucking the cells down onto the membrane rather than forcing them onto the membrane from a syringe. You will need to try several dilutions of cell suspension since too many cells could pile up and create a mess. After filtering the cells, you would then apply the various solutions (fix, wash, dehydrate) by passing through the filter. In the end, you would remove the filter and transfer it into the CPD device (keeping it wet, of course). Some holder will be needed to help keep the filters from flip flopping in the CPD device and to maintain the proper orientation (i.e., keep track of which side the cells are on). Mount the filter on the stub (double sticky tabs), coat with metal and examine in the SEM.
Good luck,
JB
} Here I am once again, hat in hand to ask for your help. This time } I have someone who wants to do SEM on a cell line grown in suspension (they } are completely non-adherent, so we can't grow them that way). What should } we do in terms of fixation, etc. and then how do we stick them to the stub? } Do we try to make them stick to something first & then run them up? Do } you run them up first & then stick them to something? Do you run up to } them & say "Stick 'em up"? } As you can tell, this is a new one to me. I've process little } things by wrapping them in lens tissue & then running the up & then shaking } the sample onto the sticky carbon dots. I don't think that procedure will } work for these because these cells are much tinier. } I thank you in advance for any suggestions you can give me. And I } thank you for all the help and advice you've given me in the past.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Does anyone have a reference for nuclear microclefts in the journal "Ultrastructural Pathology"? I'm sure I read about them in this journal three or four years ago but cannot find the reference now despite a Medline search and a look through our journals. Nuclear microclefts are sometimes found in small cell carcinomas. I am trying to establish how specific this observation is in the setting of tumour diagnosis.
Thankyou.
John Brealey, EM Unit, The Queen Elizabeth Hospital, Adelaide, South Australia.
Paula Sicurello wrote: =============================================== Here I am once again, hat in hand to ask for your help. This time I have someone who wants to do SEM on a cell line grown in suspension (they are completely non-adherent, so we can't grow them that way). What should we do in terms of fixation, etc. and then how do we stick them to the stub? Do we try to make them stick to something first & then run them up? Do you run them up first & then stick them to something? Do you run up to them & say "Stick 'em up"? As you can tell, this is a new one to me. I've process little things by wrapping them in lens tissue & then running the up & then shaking the sample onto the sticky carbon dots. I don't think that procedure will work for these because these cells are much tinier. ============================================== We have had over the years heard good reports from people who have filtered onto silver membrane filters (they cost about 3X that of a normal polymer membrane filter), the cells are kept wet, and then processed for critical point drying. They are then CPD'd and looked at by SEM. Because of the highly conductive nature of the substrate, much less gold, if any is then required, an advantage for particularly fine structured or heat sensitive samples.
A second approach, but one that I am not as enthusiastic about, but one which will work just the same, is the use of microporous specimen capsules , which have a pore size that is as small as 30 um. You can CPD the cells right in the capsules, and then when dry, with a razor blade, you can slit open the capsules, put on your conductive coating, and examine the cells in situ, right on the surface of the now slit open microporous specimen capsule
Dear Everet } } We are interested in using the SEM to measure the concentration of } flyash in the atmosphere that has come from coal-fired power plants. } One of the issues involved is being able to distinguish particles from } power plants from particles originating at other sources. We would } appreciate any information people might have on the identification of } flyash and determination of its origin. } Everett Ramer } Federal Energy Technology Center
I did a lot of SEM on Flyash some years ago which originated from different power plants. Thy were always perfect spheres. I kept some for atigmatism correction test samples. By doing EDS we were able to trace them back to the original power plants by comparing trace elements which were present in the Flyash Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
I have just heard of an elderly TEM in full working condition which is available free to anyone who is prepared to take it away. It is a Siemens Elmiskop 102 complete with Siemens image intensifier, and has had both HT cables replaced relatively recently. The instrument is installed in the north of England.
If anyone is interested, please contact me directly.
My only commercial interest is that as an ex-Siemens TEM engineer, I would be pleased to assist in the dismantling etc. if required.
Paula, Cells will adhere to coverslips or silica chips treated with poly-L Lysine (Sigma Aldrich) or suspended cells can be caught using a 0.2um polycarbonate filter. Cells should be fixed following either an incubation on the treated substrate to allow for attachment or immediately after filtering. I open the filter holder and transfer the filter to a specimen processing holder filled with fixative (EMS - cat.# 70186) Poretics (800-922-6090 ) is a good source for the 13mm Swinney syringe holders and the track etch filters. They also have a helpful specialist in Charlotte Hargrave. Rosemary
a German company called Maerzhaeuser sells motorized stages for the Olympus BH-2. Models 100 through 150 fit this Microscope. If you need more information, please contact me at my email address below.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS fax: (303) 234-9271 email: info-at-soft-imaging.com
************************************************************************ ********************************** Disclaimer: As we sell image processing systems including this stage, I do have a commercial interest in this. ************************************************************************ ******************************************************* } } ---------- } From: Shalvoy, Richard[SMTP:rbshalvoy-at-corp.olin.com] } Sent: Thursday, October 29, 1998 2:36 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Need computer controlled microscope stage } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am planning to inject an antibody conjugated with ultrasmall gold particles into a living mouse, fix by cardiac perfusion, and embed the tissues for examination by both LM and TEM. The gold particles will be silver enhanced on the sections as even our EM studies are done at rather low magnification. In the past I have done this with antibodies conjugated to 125-I, then fixed with glutaraldehyde and embedded in Epon for autoradiographic exposure. I have also fixed with paraformaldehyde and embedded in Unicryl with polymerization by UV for immunogold labelling. I am mainly interested in bone and thus want to see if decalcification with EDTA is interfering with the immunogold label. Does anyone have any opinions as to which fixative/embedding resin would be best for this type of experiment?
TIA,
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-med.mcgill.ca
Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: from rly-za05.mx.aol.com (rly-za05.mail.aol.com [172.31.36.101]) by air-za01.mail.aol.com (v51.9) with SMTP; Thu, 29 Oct 1998 09:41:11 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by rly-za05.mx.aol.com (8.8.8/8.8.5/AOL-4.0.0) with SMTP id JAA11923; Thu, 29 Oct 1998 09:38:39 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id IAA21636 for dist-Microscopy; Thu, 29 Oct 1998 08:12:17 -0600 Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id IAA21633 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 29 Oct 1998 08:11:46 -0600 } From: "Mriglermas-at-aol.com"-at-sparc5.microscopy.com Received: from imo11.mx.aol.com (imo11.mx.aol.com [198.81.17.1]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id IAA21625 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 29 Oct 1998 08:11:34 -0600 Received: from Mriglermas-at-aol.com by imo11.mx.aol.com (IMOv16.10) id NZJHa10441 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 29 Oct 1998 09:21:34 -0500 (EST) Message-ID: {ea1bb27.363879ee-at-aol.com}
Our group currently has a JEOL FX 2000 TEM available for sale to anyone interested. The system includes a cold and hot stage, PC driven X-ray system, and STEM attachment. Please respond if interested.
M.W. Rigler, Ph.D. MAS, Inc. Suwanee GA 770-866-3218
Paula, For SEM studies of cells in suspension fixed with glutaraldehyde or frozen-freeze-substituted, we attach them to silane treated (1% silane from Aldrich in acetone for a few h in 50deg.C ) and carbon coated glass or mica chips. If they are conducitively stained, they do not require coating. The most important part of the procedure is to rinse off all the non-linked aldehyde groups from the cells and free silane from the surface of the chips. We have developed this procedure with Hans Ris for SEM studies of human blood normal and leukemic cells in which we were recovering ~95% of the cells. I will be happy to guide you through, provide more details, or literature (containing also descriptions of our attempts to use other methods) if within a sphere of your interests. Marek.
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It was recently stated on the list server that the Diatome Semi and the Histo are mostly the same, and that a Histo will do the same job as a Semi. This is not true at the TEM level. The semi knives are more highly polished and create a much smoother (higher resolution) surface on epoxides. The Diatome Semi is wonderful knife for thin sectioning because one can occasionaly take a one micron section to view (ultras can not be used like this). The Diatome Histo is a marvel for thick sections. We have 22 thousand dollars worth of Diatomes. We start students thin sectioning on a Histo with the understanding that these sections are not publication quality. I don't own stock in Diatome. Wish I owned the company! Bye, Hildy
Help! Are there any histology people out there who know the whereabouts of a German company called MED'LASS? We have a tissue processor made by them about 12-15 years ago which uses acetone to dehydrate and a vacuum chamber to infiltrate. It was called the Paraffinator. To my knowledge, there are only a handful of these instruments in the U.S. and I've been unsuccessful in trying to contact the manufacturer. ANY information would be helpful. Thanks, Bob Santoianni Emory University Hospital Atlanta, Georgia robert_santoianni-at-emory.org Tel. 404-712-4874
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On Fri, 30 Oct 1998, Alexander Mironov Jr. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Dear microscopists, } We are planning to buy some diamond knives and I would like to know your } impressions of knife performance (what is the difference between Diatome, } Drukker, Pelco, DDK etc.). We need standard ultramicrotomy knives for } cutting epon embedded biological material and cryo dry ultramicrotomy } knives. Personally, I like Diatome and dislike DDK. } Did anybody see the difference in cutting and durability between 45=B0 an= d } 35=B0 knives?=20 } Which boat for standard knife is preferable? What is the purpose of boats } with inclined cavity? } Any experience is wellcome. } =20 } ___________________________ } Dr. Alexander A. Mironov Jr. } Unit of Morphology } Dept. of Cell Biology and Oncology } Consorzio Mario Negri Sud } Via Nazionale, S.Maria Imbaro (Ch) } 66030 Italy } =20 } Tel. 0872-570-332 } Fax 0872-578-240 } E-mail: amironov-at-cmns.mnegri.it } =20 } =20 } =20 } =20 I have used DDK, Dupont, Diatome, others. For the last ten years we have invested only in Diatome. For the last seven years we have about 22 thousand dollars worth of assorted Diatome knives. We have Never had a bad one, never had one poorly resharpened (I long ago quit testing them when they came back to us), never had one that wore out quickly. Furthermore, Diatome USA has a laboratory set up. In case you have trouble with your embedding or materials, you can send them some of your tissues with your knife, and they will probe the situation for you and give you advice. I have not used this service, but I know from others that this service is fine. We have gotten excellent advice also on solving problems with our microtomes which was affecting the knives. I don't own stock in Diatome. Wish I did.
I'm interested in obtaining information about all commercially available cathodoluminescence detector systems. This includes both stand-alone and SEM add-on systems. I would like to generate a list of vendors that manufacture CL instrumentation.
Many thanks in advance. Vendors are welcome to contact me.
Best regards,
Angela
--------------------------------------------- Angela V. Klaus
Manager - Core Microscopy Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
Hello. My recomandation for knifes would be to buy Drukker knives! It seems like most of you prefere the Diatome knife. At my lab we have during the last few years changed from Diatome to Drukker knifes, both for resin embedded material (Epon and Lowicryl)and for cryosections. All of us (we are seven people using this knives routinely) all think that the Drukker knives are better than Diatome, and for the two of us making cryosections most of the time it sure made sectioning easier.
I have a collaborator who wants to produce some EM samples using a = spraying procedure. The particles which are produced are quite large (up = to 50=B5m in diameter).=20
I know how to handle suspensions, but these particles are buoyant and so = will not settle. (Sorry to be so vague about details, but the work is = confidential in nature.)
I remember, from my undergrad EM courses, many years ago, that samples can = be sprayed using a =22nebulizer=22, but I can=27t seem to locate one to = try.
Can anyone help me locate a nebulizer, or suggest other options for = producing these samples?
As always, thanks for your help.=20
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
--- --- --- -- -- -- --- --- --- Richard J. Dudley (rdudley+-at-pitt.edu) Research Specialist V Dept. of Cell Biology and Physiology University of Pittsburgh http://www.cbp.pitt.edu ---} search BIONET archives at http://www.bio.net {---
In another week or so I'll be leaving the art and science of scanning electron microscopy for a new career as a quality manager in our automotive lighting division. Before I go, I'd like to thank you, the worldwide community of microscopists in this forum, for your generous offerings of free advice, tips, and insights into our craft. You have allowed me to become a better microscopist and a better service provider for my customers. This is a debt I cannot repay other than to say that I will carry your spirit of openness and collaboration into my new job.
I'd also like to thank our friendly neighborhood system administrator for his uncanny ability to remain friendly in this neighborhood. I know I wouldn't have the patience to do what he does on our behalf.
Best wishes to all of you,
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com http:///www.sylvania.com
Dear colleagues: We are recently asked to process mouse sperm for SEM. Do any of you have specific protocol for such specimen at hand? Or can we process the sample as ordinary cell suspension? Thanks in advance to any help. Yuhui Xu, MD EM Core, DFCI
I would like to thank all who shared their experience about the performance of diamond knives and told me the address of Agar. Thank you.
___________________________ Dr. Alexander A. Mironov Jr. Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
In a message dated 98-11-05 08:22:38 EST, allanwojtasp-at-em.agr.ca writes:
{ { I have a collaborator who wants to produce some EM samples using a spraying procedure. } }
Hi Paula,
I have used a nebulizer for just such purposes, but like you, I can't for the life of me remember where we got it. I recall vaguely that it was from a medical supply firm. The word "Vaponefrin" sticks in my mind, but this may be a trade name for an inhalable form of epinephrine rather than the name of the nebulizer.
But I have also used an artist's airbrush with good results. The airbrush is nice because you can regulate the air pressure and the size of the droplets which are produced. Plus, you can get one easily at your local arts and crafts supply store.
There are also stainless steel ultrasonic nebulizers which are hand-held devices (not the large ones for use in homes), but they tend to be quite expensive.
I would go with the airbrush if you can't find a nebulizer.
Good luck, hope this helps!
Cheers,
Bob **************************************** Robert (Bob) Chiovetti Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives / Systems Integrators / Analog & Digital Imaging *****************************************
We at Ladd Research, like many of our competitors, sell nebulizers. In our case it is catalog number 23625. If you are interested we could get you specs and a price.
Thabks,
JD Arnott
Paula Allan-Wojtas wrote: } } } Hi, all, } } I have a collaborator who wants to produce some EM samples using a spraying procedure. The particles which are produced are quite large (up to 50µm in diameter } } I know how to handle suspensions, but these particles are buoyant and so will not settle. (Sorry to be so vague about details, but the work is confidential in } } I remember, from my undergrad EM courses, many years ago, that samples can be sprayed using a "nebulizer", but I can't seem to locate one to try. } } Can anyone help me locate a nebulizer, or suggest other options for producing these samples? } } As always, thanks for your help. } } Paula. } } Paula Allan-Wojtas } Food Microstructure Specialist } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca }
--
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
We have used and inexpensive air-brush in the past. You can find them at= any hobby store and they are much more adjustable than a nebulizer. We = use N2 gas to operate the air-brush as it is cheap and basically inert. Greg
Paula Allan-Wojtas wrote:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } } Hi, all, } } I have a collaborator who wants to produce some EM samples using a spra= ying procedure. The particles which are produced are quite large (up to 5= 0=B5m in diameter). } } I know how to handle suspensions, but these particles are buoyant and s= o will not settle. (Sorry to be so vague about details, but the work is c= onfidential in nature.) } } I remember, from my undergrad EM courses, many years ago, that samples = can be sprayed using a "nebulizer", but I can't seem to locate one to try. } } Can anyone help me locate a nebulizer, or suggest other options for pro= ducing these samples? } } As always, thanks for your help. } } Paula. } } Paula Allan-Wojtas } Food Microstructure Specialist } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
-- =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D
Paula, 50 micron samples are a little large for EM. Why aren't you = using LM? Are these samples dry or already in liquid? Are they boyant due to = density or surface tension? Spraying samples or other techniques must be used = with caution as unwanted classification can be difficult to minimize = especially if your size distribution is large. Is your goal size distrubition? = Little can be suggested without knowing a lot more about the sample. Russ
-----Original Message----- } From: Paula Allan-Wojtas [mailto:allanwojtasp-at-em.agr.ca] Sent: Thursday, November 05, 1998 8:04 AM To: microscopy-at-sparc5.microscopy.com
Hi, all,
I have a collaborator who wants to produce some EM samples using a = spraying procedure. The particles which are produced are quite large (up to = 50=B5m in diameter).=20
I know how to handle suspensions, but these particles are buoyant and = so will not settle. (Sorry to be so vague about details, but the work is confidential in nature.)
I remember, from my undergrad EM courses, many years ago, that samples = can be sprayed using a "nebulizer", but I can't seem to locate one to try.
Can anyone help me locate a nebulizer, or suggest other options for producing these samples?
As always, thanks for your help.=20
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
i process quite a bit of mouse sperm for SEM. I usually have the investigator wash it and put fixative (2% glutaraldehyde in cacodylate buffer) and then bring it to me. I let the cells fix for about two hours in the fridge. Then I poly-L-lysine (1%) coat 12mm round coverslips for about 10 minutes. I wash off the poly-L-lysine with distilled water, add a drop of sperm in fixative and let that sit for 10 minutes. Then I wash with 25% ethanol and dehydrate the coverslips to 100% ethanol. They can be critical point dried, coated and viewed in the sem. This works well for red blood cells too.
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
Well, we all know microscopy is expensive, but worth it. However, it is not always easy to convince the powers-that-be. Now we are being asked to carry some of the weight by doing commercial work. I seem to remember a thread about this some time ago and the problems of the University of Hawaii. Does anyone remember the problems? Does anyone at another university have any ideas about doing commercial work? Thanks for any help you might give me. Joyce Chicago State University
On Thu, 05 Nov 1998 08:04:26 Paula Allan-Wojtas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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On Thu, 05 Nov 1998 08:04:26 Paula Allan-Wojtas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Paula. Ted Pella sells nebulizers. You could make one by putting a T in a stopper on a glass flask. Hook compressed gas to one side, and a pipette to the other; you need a small diameter tube to extend from the inside of the Pipette to the liquid in the flask. -be careful -a
On Thu, 05 Nov 1998 08:04:26 Paula Allan-Wojtas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Dear Paula, If you can produce the particles, just dry a drop or two of suspension onto a polished graphite-covered stub or a stub with a C-impregnated sticky tab and examine, or gold-coat and examine. A nebulizer is just any device that forces a liquid through an orifice to break it into small drops. The smaller the orifice or the greater the force, the smaller the drops. You wrote:
} Hi, all, } } I have a collaborator who wants to produce some EM samples using a spraying procedure. The particles which are produced are quite large (up to 50=B5m in diameter).=20 } } I know how to handle suspensions, but these particles are buoyant and so will not settle. (Sorry to be so vague about details, but the work is confidential in nature.) } } I remember, from my undergrad EM courses, many years ago, that samples can be sprayed using a "nebulizer", but I can't seem to locate one to try. } } Can anyone help me locate a nebulizer, or suggest other options for producing these samples? } } As always, thanks for your help.=20 } } Paula. } } Paula Allan-Wojtas
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
I am setting a new TEM lab in my institute and need many things. One of them is carbon rod sharpener to make 3 mm diameter carbon rod with a cylindrical tip. I found that the commercial one was very expensive relatively. Could anybody suggest how I can get a generic sharpener at a low price? Any other suggestion is also appreciated.
Jondo Yun Department of Inorganic Materials Engineering Kyungnam University 449 Weolyeong-dong Masan, 631-701 Korea 82-551-249-2697 (office) 82-551-248-5033 (fax) 82-551-249-2692 (department office) 82-551-249-2719 (laboratory) 82-551-249-2564 (EM lab) email: jdyun-at-hanma.kyungnam.ac.kr
Be extremely cautious here. If any of your instruments were purchased using NSF funding and/or were purchased DUTY-FREE you will likely be violating various rules concerning the use of this equipment for commerical organizations. NSF has specific rules (NSF Notice 91) regarding instruments purchased with their funds (or at least they did last time I checked). Similiarly if you buy equipment duty free the forms (again the last time I filled one out) specifically require you to testify that it will not be used to be for commerical work for others.
There are alot of commerical "service" organizations that have paid full freight to buy their instruments and these RULES are made to protect their interests.
There is also a new law just passed in Congress S 314 which requires the governmental organizations (not defined as far as I know) to document any commerical activities and justify those which they perform. The intent appears to insure that the governmental resources do not compete with the commerical sector. It's a very narrow line and very easy to cross. I think at the moment that S314 is directed at Federal Employee's, but it could conceivably also be applied to anyone with Federal $$$.
I guess I'm waving a big red flag and telling you to tread carefully. In any event you had better point this out to your powers that be and have your legal eagles at CSU investigate all your potential liabilities. You could conceivably have to pay back the DUTY FREE fees or loose all future DUTY FREE purchasing ability.
The Santa Clara Valley Chapters of ASM and IEEE Reliability Society, with generous support from FEI Company, Micrion Corp, Schlumberger, and Nissei Sangyo America present:
NANOSCALE CHARACTERIZATION WITH THE FOCUSED ION BEAM Speaker: Prof. Robert Hull Department of Materials science and Engineering University of Virginia
ABSTRACT: Nanoscale characterization techniques using the focused ion beam (FIB) instrument will be reviewed. Dr. Hull will describe FIB-based techniques (either stand-alone, or in conjunction with transmission electron microscope imaging) for stress mapping in crystalline structures, dopant mapping in semiconductors, three dimensional image reconstruction, and ultra-high resolution secondary ion mass spectroscopy maps and volume reconstructions. He will also summarize additional techniques described in the literature, including FIB-induced optical emission spectroscopy and voltage contrast imaging. Finally specimen modification and damage artifacts created by the FIB beam will be discussed.
BIOGRAPHICAL SKETCH Robert Hull is an Associate Professor, and Doris and Heinz Wilsdorf Distinguished Research Chair, in the Department of Materials Science and Engineering at the University of Virginia. Prior to joining UVA he was a member of Technical Staff in the Physics Research Division of Bell Laboratories for seven years. He has authored and co-authored over one hundred and fifty papers in the fields of electronic materials, epitaxial growth, and applications of focused ion and electron beams. He has also given over fifty invited presentations at national and international conferences in these fields. He is on the editorial board of several major journals, and has edited several proceedings and reference volumes. In 1997 he was the President of the Materials Research Society, the leading international society in the field of materials science and engineering.
TIME AND LOCATION: November 11 at David's Restaurant at the Santa Clara Tennis and Golf Club, at 5151 Stars & Stripes Drive in Santa Clara, CA (95054). This is just east of the Santa Clara Convention Center. Dinner choice of: London Broil or Seafood Brochette served with lemon butter sauce or a Vegetarian (Pasta Primavera) plate. Social at 6:00 p.m., 6:45 Dinner and 8:00 p.m. Talk
Cost: ASM/IEEE Members $16, Students $8 and Guests $18 (with an additional two dollars if no RSVP the Monday before the event (i.e, Nov 9th) Reservations: Brock Hinzmann 650-859-4350 or email IEEE Santa Clara Valley Chapter Reliability Society contact David Su (davidsu-at-aol.com). Please include choice of meal!
At 09:51 05.11.98 EST, you wrote: } What is it that you base your decision on. Do you get a kickback from } Drukker? }
No kickback from Drukker! But I'm one of those who actually have had the oprtunity to compare the kniver, and like the Drukker ones. If that is not good enough I can't see the value of asking about peoples opinions! By the way; who is actually behind the SGKCCK adress??? RO
Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso MH-Breivika N-9037 TROMSO NORWAY
Dear all, I am passing this message on for a colleague who is wondering about purchasing a better scanner, specifically for TEM plate negatives. He already has a UMAX Astra 1200S with transparency adaptor but this does not cope with high optical densities, or with enlarging small areas greatly. We use two different sizes of plate film: one just smaller than 3 1/4" by 4" and the other 2 1/2" by 3 1/2". What would you recommend, a high end flatbed with transparency adaptor or a specialist negative scanner? Which models have worked well for you. He would prefer it to be Mac compatible.
Thanks
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, or: ianmaclaren-at-hotmail.com Birmingham B15 2TT, http://web.bham.ac.uk/I.MacLaren England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
We have dealt with this issue extensively. You can always try to do this through your office of sponsored programs or wahatever they call it there. The outside person can give you a grant or gift to do the work. We then put the money in a University Foundation Account. You may also be able to enter into a contract for research or services . What if any overhead the university might take needs to be negotiated. We generally no longer operate in that manner since we have been reorganized as an auxiliary business. That is we recharge our internal customers and are able to serve outside cutomers as resources permit. This is a business incidental to an educational activity and is looked on much as the university bookstore is. Laws and regulations vary from state to state. Fell free to contact me for more info. I also got into Federal Cost Accounting morass quite deeply and can discuss that with you as well.
Greg At 03:19 PM 11/05/1998 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks for all of you who responded with locations for nebulizers, and other options for spraying the samples. Some we have tried already, and some we will try within the next month.
I can give you a little more information, and ask for help with a slightly different problem this time. We are also looking for ways of producing the sample, not just depositing it on EM supports (grid or stub). The collaborator tells me that they are now able to make the samples quite abit smaller in diameter using thier traditional methods.
The reason why we are using EM at all is that we are studying the microstructure of the sample and changes in it. The collaborator uses LM as a quick check after each modification is made.
I remember seeing a method where a grid or some support is placed in a centrifuge tube with the sample, and the sample is "spun" onto the support. Does anyone have a reference for this one?
Thanks very, very much!
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
1) histo knives - you are only partly correct, Hildegard. In terms of materials science 'hard' materials sectioning, histo knives can indeed furnish publication quality micrographs in the sense that the sectioning process itself inevitably induces so much deformation damage in our crystalline materials that the 'rippled' surface produced by the histo is of little consequence relative to the desired information from the section. (I call it 'continuous knife marks' because that is what it looks like). No less a personage than Prof. Helmut Sitte in Germany agrees with our thinking that this may be due to a serrated nature for the knife edge that might just serve the purpose of a self-sharpening effect of sorts. We have tested histo knives from both Diatome and DDK on metals (some quite hard) and have observed the following (summarized also in EMSA/92, p. 294): - they produce quite good ultrathin sections (if one ignores the above marks), indeed the thinnest sections we have ever produced - {10nm in Al - were with a Diatome histo, - they produce incredibly flat sections, seemingly because the unique nature of the edge somehow counteracts the asymettric buildup of deformation that produces the dreaded curling of metallic sections (like in lathe turnings) that drives our microtomist nuts at times, - for some ultrahard metals (FE-Nd-B amorphous intermetallic particles), it was the only knife (compared to 35, 45 and 55 degree colleagues) that produced useable sections, - before you all rush out and buy histos for materials science sectioning, they also; vary quite a bit more in performance than regular knives (what do you expect for half the price?), may start to 'shred' the section as the thickness drops below 100nm, and are hopeless for cutting anything with weakly adherent interfaces (coatings and the like).
2) As per the related thread about Diatome vs Drukker, etc; we have used knives from Diatome, Drukker, Microstar and DDK over the last 15 years (all for materials science sectioning), yet I am always loathe to give any sort of public ranking or recommendations. A lot depends on the materials you are sectioning, your relationship with the knife producer, how you clean and care for your knives, what is the stage the edge at any given time, etc. Our person prefers the Diatomes (we have several), but has obtained good results from all of the others, so-----, each to your own. The important point is that, over the years, I have found most diamond knife producers to be amazingly cooperative and helpful with regard to performance questions, tips and hints, resharpening and the like. If only some of the other makers of EM accessories were so helpful!
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 623-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: HILDEGARD CROWLEY[SMTP:hcrowley-at-du.edu] } Sent: November 04, 1998 2:02 PM } To: postmessage } Subject: Re: Diatome Semi, Ultra, Histo } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } It was recently stated on the list server that the Diatome Semi and the } Histo are mostly the same, and that a Histo will do the same job as a } Semi. This is not true at the TEM level. The semi knives are more highly } polished and create a much smoother (higher resolution) surface on } epoxides. The Diatome Semi is wonderful knife for thin sectioning because } one can occasionaly take a one micron section to view (ultras can not be } used like this). The Diatome Histo is a marvel for thick sections. We } have 22 thousand dollars worth of Diatomes. We start students thin } sectioning on a Histo with the understanding that these sections are not } publication quality. } I don't own stock in Diatome. Wish I owned the company! } Bye, } Hildy } }
Could you please send this to the EM Server? Thanks.
We are looking at EM tissue processors and hope to make a purchase soon. We process primarily biological samples. Does anyone have experience with the EMP5160 manufactured by RMC? It is similar to the LKB tissue processor that we used to have but seems to have some nice modifications. If anyone has experience with this model (pros or cons), or any other suggestions, please contact me at gatlin_cindy_l-at-lilly.com or reply via server if you think others might be interested.
Thanks very much,
Cindy Gatlin Technical Associate/EM lab Eli Lilly & Co.
Ian asks ... } } } Dear all, } I am passing this message on for a colleague who is wondering about } purchasing a better scanner, specifically for TEM plate negatives. He } already has a UMAX Astra 1200S with transparency adaptor but } this does not } cope with high optical densities, or with enlarging small } areas greatly. } We use two different sizes of plate film: one just smaller } than 3 1/4" by 4" and the other 2 1/2" by 3 1/2". } What would you recommend, a high end flatbed with } transparency adaptor or a specialist negative scanner? ...
Since EM image acquisitions typically are the result of "scans" and therefore depict scan lines, I might suspect a scanner for specific film types might cause moire patterns. That is, the scanner would insist you mount the film almost orthogonally, whereas you might try to remedy moire patterns by tilting the film on a higher res flatbed ... say, 30deg, and then making it orthogonal again with your favorite image editor. Dedicated film scanners would offer the resolution you want ... I'd simply suggest you try them before you buy them.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
} Well, we all know microscopy is expensive, but worth it. However, it is } not always easy to convince the powers-that-be. Now we are being asked } to carry some of the weight by doing commercial work. I seem to } remember a thread about this some time ago and the problems of the } University of Hawaii. Does anyone remember the problems? Does anyone } at another university have any ideas about doing commercial work? } Thanks for any help you might give me. } Joyce } Chicago State University
Joyce -
The problem in Hawaii was different; it had to do with the way lab usage was being charged to federal grants. I dealt with commercial usage pre-retirement at U.C. Berkeley. There were very specific statewide university rules that covered all such use on the campus. I was allowed to provide service to outsiders only if there was no private lab in the local area offering the same service, and I was told to charge double the campus recharge rate. I supported the policy, because competition with private enterprise using grant-funded equipment really is unfair.
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
When we decided to buy a scanner for negatives we tested different scanners, from the top-end scanner used for cartography (extremely expensive and with pixel size of about 7.5 microns) down to flatbed scanners with transparency adaptor. From our tests we obtained that for TEM plates negative scanners were better and that for high resolution TEM work at least (optical) 1600 dpi were required in order to be able to enlarge the image sufficiently. We decided to buy a Polaroid SprintScan 45, which accepts large negatives. We use 2000x2000 dpi for HREM work. Optical density is about 3.4, if I'm not wrong. We are satisfied with this instrument, except for the negative holder: for 3 1/4"x4" negatives the holder is too large and some bending of the negative occurs, as it can only be held on the two short sides. However we didn't notice large image distortion. I think that I read in this list that Polaroid was preparing special holders for the negatives, but I don't know further details. Is there somebody that could answer this question? For smaller negatives an adapter exist with two magnets, but I don't know whether your negatives can be fitted in it (included with the scanner). As conclusion, during the last year I didn't use the dark room at all and I think that I will extremely seldomly use it again (only if our dye-sub printer breaks). I hope this helps.
Albert Albert Romano-Rodriguez Dept. of Electronics Faculty of Physics University of Barcelona c/ Marti i Franques, 1 E-08028 BARCELONA Spain tel: +34-93-402 11 47 FAX: +34-93-402 11 48 e-mail: romano-at-el.ub.es
We are in the early stages of evaluating a digital imaging system here (for light microscopy). I'm trying to understand the different ways that CCD cameras can be or are used to acquire color images. There seem to be several ways this is done:
* Single chip Monochrome CCD array with some type of filter in front of the camera to allow it to acquire sequential red, green & blue images and then software to "reassemble" the images into a color image. * 3 Chip camera, with each chip assigned (filtered for) red, green & blue and then software to "reassemble" the images into a color image. * Single chip CCD with some type of color mosaic "mask" on the chip to acquire the red, green & blue parts of the image and then software to "reassemble" the images into a color image. * CCD array where the image is "scanned" either by moving optical elements or moving the CCD array to acquire a high pixel count with a fairly small sensor. These would be the slowest for acquisition, but I gather they give a lot of "bang for the buck (euro)". * Others?
What are the Pros & Cons of these different types of cameras? Is there a WWW or published resource that could help me sort this out?
Our interest here is primarily in acquiring static images (not real-time video) from low light level fluorescence, DIC and/or bright field. We would like the camera to be sensitive enough for quantitation if needed. I'd also like to be able to advise others here who may have different requirements. I'm not specifically "fishing" for sales pitches (I already have plenty of glossy literature, I'm just trying to make sense out of it).
I would be happy to take replys "off-list" and post a summary.
Yours, Doug .................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
} I am passing this message on for a colleague who is wondering about } purchasing a better scanner, specifically for TEM plate negatives. He } already has a UMAX Astra 1200S with transparency adaptor but this does not } cope with high optical densities, or with enlarging small areas greatly. } We use two different sizes of plate film: one just smaller than 3 1/4" by } 4" and the other 2 1/2" by 3 1/2". } What would you recommend, a high end flatbed with transparency adaptor or a } specialist negative scanner? Which models have worked well for you. He } would prefer it to be Mac compatible. } Dear Ian, I think the best scanners are either the flat-bed (like the Perkin Elmer) or rotating Drum (like the Optronics). These are capable of 5-by-5 micrometer pixel size, in the case of the PE, and the files produced can by presented in any size needed depending on the image processing program used. The SPIDER system used here will handle both PE and Op data. The bad news is that these are running on a mainframe, and, AFAIK, not Mac compatable. There may be software out there which runs on the Mac and can read PE or Op files. Good luck. Yours, Bill Tivol
=C0=B1 =C1=B8=B5=B5 Jondo Yun wrote: } =20 } -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of Dear Jo= ndo, =20 } I am setting a new TEM lab in my institute and need many things. } One of them is carbon rod sharpener to make 3 mm diameter carbon rod wi= th a } cylindrical tip. I found that the commercial one was very expensive } relatively. } Could anybody suggest how I can get a generic sharpener at a low price? } Any other suggestion is also appreciated. } =20 Ted Pella sells a hand-held sharpener for under $100. You should check his web site for info. I am not connected with Pella except as a customer. Yours, Bill Tivol
Michael Shaffer's comment seems a bit off the mark. TEM negatives are generally not scanned images. Although some types of images (e.g., crystal lattice images) can give moire's when scanned, this effect can be avoided by appropriate choice of scanner sampling resolution. I concur with the 'try before you buy' philosophy, if the opportunity can be made.
The optical density range of high-end 12-bit grayscale scanners (36 bit color) goes as high as 3.6. Unfortunately, scanner software seldom offers exposure control. Actually, 3.6 or even (more common) 3.2 is an adequate density range for most TEM photos including diffraction patterns. Your microscopist colleague might consider controlling exposures in the microscope to avoid excessively dense negatives. There are also special film developing solutions that might be used for special situations where contrast reduction is necessary.
My personal observation of current 36-bit, } 1000 spi scanners is that they are incredibly slow and inefficient for scanning TEM negatives. Anyone contemplating a market survey should include in their evaluation the time takes to scan into Photoshop including time the scanner takes calibrating itself before each scan.
Don't know if you saw my earlier posting to the listserver concerning scanning TEM negatives. The essence was that the 'negative film scanning mode' gives poor grayscale results for high-contrast TEM negatives because of a built in contrast correction feature. The scanned images become posterized. The work-around is to always scan in the positive transparency mode and invert the image contrast with other controls.
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA USA
---------- From: shAf Sent: Friday, November 6, 1998 5:01 PM To: Ian MacLaren; Microscopy-at-Sparc5.Microscopy.Com Cc: g.r.millward-at-BHAM.AC.UK Subject: RE: Scanning negatives
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Ian asks ... } } } Dear all, } I am passing this message on for a colleague who is wondering about } purchasing a better scanner, specifically for TEM plate negatives. He } already has a UMAX Astra 1200S with transparency adaptor but } this does not } cope with high optical densities, or with enlarging small } areas greatly. } We use two different sizes of plate film: one just smaller } than 3 1/4" by 4" and the other 2 1/2" by 3 1/2". } What would you recommend, a high end flatbed with } transparency adaptor or a specialist negative scanner? ...
Since EM image acquisitions typically are the result of "scans" and therefore depict scan lines, I might suspect a scanner for specific film types might cause moire patterns. That is, the scanner would insist you mount the film almost orthogonally, whereas you might try to remedy moire patterns by tilting the film on a higher res flatbed ... say, 30deg, and then making it orthogonal again with your favorite image editor. Dedicated film scanners would offer the resolution you want ... I'd simply suggest you try them before you buy them.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Nestor's caution is fully justified. His description is right on the mark, and the rules have just been re-issued as Important Notice number 122 of 1998 (see http://www.nsf.gov/pubs/1998/iin122/iin122.txt).
Caroline was generous. We charge commercial customers three times our academic rate - but the academic rate is the same for MIT and any other academic user.
The microscope is currently in use. It has some good days and some bad days and it will be available around Thanksgiving. It's free!! (If your willing to haul it away.) The SEM resides in Easton PA.
If you think you might be interested, give me a call: Nan Laudenslager Specialty Minerals, Inc. (610) 250-3094
} Some diamond knife comments: } } 1) histo knives - you are only partly correct, Hildegard. In terms of } materials science 'hard' materials sectioning, histo knives can indeed } furnish publication quality micrographs in the sense that the sectioning } process itself inevitably induces so much deformation damage in our } crystalline materials that the 'rippled' surface produced by the histo is of } little consequence relative to the desired information from the section. (I } call it 'continuous knife marks' because that is what it looks like). No } less a personage than Prof. Helmut Sitte in Germany agrees with our thinking } that this may be due to a serrated nature for the knife edge that might just } serve the purpose of a self-sharpening effect of sorts. We have tested } histo knives from both Diatome and DDK on metals (some quite hard) and have } observed the following (summarized also in EMSA/92, p. 294): } - they produce quite good ultrathin sections (if one ignores the above } marks), indeed the thinnest sections we have ever produced - {10nm in Al - } were with a Diatome histo, } - they produce incredibly flat sections, seemingly because the unique nature } of the edge somehow counteracts the asymettric buildup of deformation that } produces the dreaded curling of metallic sections (like in lathe turnings) } that drives our microtomist nuts at times, } - for some ultrahard metals (FE-Nd-B amorphous intermetallic particles), it } was the only knife (compared to 35, 45 and 55 degree colleagues) that } produced useable sections, } - before you all rush out and buy histos for materials science sectioning, } they also; vary quite a bit more in performance than regular knives (what do } you expect for half the price?), may start to 'shred' the section as the } thickness drops below 100nm, and are hopeless for cutting anything with } weakly adherent interfaces (coatings and the like). } } 2) As per the related thread about Diatome vs Drukker, etc; we have used } knives from Diatome, Drukker, Microstar and DDK over the last 15 years (all } for materials science sectioning), yet I am always loathe to give any sort } of public ranking or recommendations. A lot depends on the materials you } are sectioning, your relationship with the knife producer, how you clean and } care for your knives, what is the stage the edge at any given time, etc. } Our person prefers the Diatomes (we have several), but has obtained good } results from all of the others, so-----, each to your own. The important } point is that, over the years, I have found most diamond knife producers to } be amazingly cooperative and helpful with regard to performance questions, } tips and hints, resharpening and the like. If only some of the other makers } of EM accessories were so helpful! } } } Tom Malis } Group Leader - Characterization } Materials Technology Laboratory } Natural Resources Canada (Govt. of Canada) } 568 Booth St., Ottawa, Canada } ph. 613-992-2310 } FAX 623-992-8735 } email: malis-at-nrcan.gc.ca } } } } } ---------- } } From: HILDEGARD CROWLEY[SMTP:hcrowley-at-du.edu] } } Sent: November 04, 1998 2:02 PM } } To: postmessage } } Subject: Re: Diatome Semi, Ultra, Histo } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi, } } } } It was recently stated on the list server that the Diatome Semi and the } } Histo are mostly the same, and that a Histo will do the same job as a } } Semi. This is not true at the TEM level. The semi knives are more highly } } polished and create a much smoother (higher resolution) surface on } } epoxides. The Diatome Semi is wonderful knife for thin sectioning because } } one can occasionaly take a one micron section to view (ultras can not be } } used like this). The Diatome Histo is a marvel for thick sections. We } } have 22 thousand dollars worth of Diatomes. We start students thin } } sectioning on a Histo with the understanding that these sections are not } } publication quality. } } I don't own stock in Diatome. Wish I owned the company! } } Bye, } } Hildy } } } } } I have never been involved with materials science. My remarks only apply to biological materials.
DVC Company has an answer to your need for a decent high resolution camera that others reading this might need. DVC has introduced this at the Vision Show in San Jose last month and at the Photonics East in Boston last week and now.....at the Neuroscience show in LA at our DVC booth in the 8XX area starting this Sunday. DVC does the whole system and the color info is done in the host/ 400Mhz computer which we supply along with the Epix frame grabber that is only $995.
See our web http://members.aol.com/dvcco
We have a 1300 x 1030 pixel digital camera that uses a single sensor with a color Bayer pattern. The singel sensor offers in effect } 700 TV Lines square from our model DVC1300C and in monochrome at } 1000 TV Lines Square. Using a single sensor offering 12 frames / sec is much better if real time 30fps is not needed because we offer the first camera to do this with 58-60dB S/N !!! You have no color shift as with 3 chip CCD cameras with a price of $4995. You also have a higher vertical resolution as stated above / square pixels, and no problem with ( color shift ) which comes from misalighnment of the 3 chips on a prism situation and the cost......10K plus....and bulky also. Our unit weigh's less than one pound. You can go with a bulky expensive $1500-2500 attachable LCD filter on the monochrome version of the camera but........you loose 30% sensitivity.....and only gain perhaps 200 TV Lines over our 700TV Line unit.... You should check out our web on this and visit us at Neuroscience. For those who feel offended from learning something new, especially from the camera manufacturer..... well excuse me ! With all due respect for the others that do not. Regards,
Rich
Richard Klotsche DVC Company/ San Diego 619-444-8300 619-444-8321-fax
In a message dated 98-11-06 14:17:40 EST, you write:
{ { Subj: Color CCD Camera ?s Date: 98-11-06 14:17:40 EST From: doug-cromey-at-ns.arizona.edu (Doug Cromey) To:
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Microscopy Listers,
We are in the early stages of evaluating a digital imaging system here (for light microscopy). I'm trying to understand the different ways that CCD cameras can be or are used to acquire color images. There seem to be several ways this is done:
* Single chip Monochrome CCD array with some type of filter in front of the camera to allow it to acquire sequential red, green & blue images and then software to "reassemble" the images into a color image. * 3 Chip camera, with each chip assigned (filtered for) red, green & blue and then software to "reassemble" the images into a color image. * Single chip CCD with some type of color mosaic "mask" on the chip to acquire the red, green & blue parts of the image and then software to "reassemble" the images into a color image. * CCD array where the image is "scanned" either by moving optical elements or moving the CCD array to acquire a high pixel count with a fairly small sensor. These would be the slowest for acquisition, but I gather they give a lot of "bang for the buck (euro)". * Others?
What are the Pros & Cons of these different types of cameras? Is there a WWW or published resource that could help me sort this out?
Our interest here is primarily in acquiring static images (not real-time video) from low light level fluorescence, DIC and/or bright field. We would like the camera to be sensitive enough for quantitation if needed. I'd also like to be able to advise others here who may have different requirements. I'm not specifically "fishing" for sales pitches (I already have plenty of glossy literature, I'm just trying to make sense out of it).
I would be happy to take replys "off-list" and post a summary.
Yours, Doug .................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
We have been using an RMC tissue processor 5160 for several years. Our processor may be a bit different than the one currently sold, as it not only is designed to keep the reagent vial in which the specimens reside at a specific temperature, it will also pre-cool the solution vial in which the samples are about to be placed as well. I believe that the newer model eliminated temperature control on the "next" vial, which in my mind means that all the solutions should be used at ambient temperature to avoid significant temperature fluctuation within the samples.
There are several "quirks" with the use of the machine which you will learn over time (or contact me to avoid reinventing the wheel). For example, we do not use the reagent vial caps unless necessary, since sometimes the caps are not secured to the vials during refitting (an alignment problem). The caps then fall into the mechanics and cause a jam. We also put small holes in the caps above volatile solutions (propylene oxide, 100% ethanol, acetone) since evaporation will cause the lids to lift up and become askew, causing an alignment problem which again causes the caps to fall into the machinery.
We have had some difficulties obtaining expendable, including specimen holders and particularly reagent vials. The design of the vials has been improved over time (they have thicker walls and don't warp as easily...warping caused the specimens to get stuck as they were transferred from one vial to the next, eventually causing the samples to be lost or confused with others).
Overall, despite the above comments, I really like the processor. Although it takes about 30 minutes to set up, it eliminates countless hours spent in the fume hood transferring solutions. Exposure to organic solvents, carcinogens and fixatives is much less likely. The machine allows overnight processing of tissues. We reuse many of our reagent vials (not the ones containing epoxies or OsO4) so it is not too expensive to use. It does use more volume of reagents than you may be accustomed to. We routinely use 10 to 18 ml in each of the 22 reagent vials, depending on how many samples we are processing (up to 12). Although I have done so, I am nervous to use the processor for tissues which are irreplaceable. If I do use it for these tissues, I prefer to do so when either myself or my technician is available to visit the machine occasionally to observe its performance.
I'd be happy to talk more with you over the phone.
Best wishes,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
} The optical density range of high-end 12-bit grayscale scanners (36 bit color) } goes as high as 3.6. Unfortunately, scanner software seldom offers exposure } control.
While I concur with you about most of your message, I am surprised by this statement. We have several scanners on our floor and all of them have software which permits control of density and contrast, both manually and via presets. One can shift the grayscale 'window'quite easily and extract information even from quite dense negatives.
Ian, I compared some scanners for TEM negatives a few months ago. It all depends on the level of quality needed and how much enlarging you mean . 1200 dpi was sufficient for low mag. stereology and morphometry, and contrasty high mag. such as colloidal gold and dense histochemical deposits. At 2400 dpi the film's silver grains could be distinguished at high enlargements, suggesting nearly lossless scan. The 2400 dpi scans were suitable for low contrast, high mag. work and enlargments. The scanner everyone really loved was the Imascan, by Imacon (www.imacon.dk). It is essentially a small drum scanner with a flexible magnetic holder for positives or negatives that is very easy to use. At 5760 dpi, 14 bit and a Dmax of of 4.1, the images it produced could be enlarged to give distinct view of silver grains. These files did amount to about 30 Mb for a monochrome 3 1/4x4 negative. I put some extremely dense negatives through it with excellent results. the price on the Imascan is probably steeper than you want, US$16,000. The main EM users in our group are contemplating a joint purchase. The Imascan is driven by its own software for Mac or Windows and connects to either platform via SCSI port. I tested it with a 233 MHz G3 and found the scan times tolerable unless one was doing RGB at 2400 dpi or higher. then the 3 passes added up. 2 other scanners we are considering as an alternative to the Imascan are the LinoColor Ultra Sapphir and the Agfa Duoscan. They seem to be better able to get closer to their stated Dmax than some of the other brands.
Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
On Fri, 6 Nov 1998, Ian MacLaren wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } I am passing this message on for a colleague who is wondering about } purchasing a better scanner, specifically for TEM plate negatives. He } already has a UMAX Astra 1200S with transparency adaptor but this does not } cope with high optical densities, or with enlarging small areas greatly. } We use two different sizes of plate film: one just smaller than 3 1/4" by } 4" and the other 2 1/2" by 3 1/2". } What would you recommend, a high end flatbed with transparency adaptor or a } specialist negative scanner? Which models have worked well for you. He } would prefer it to be Mac compatible. } } Thanks } } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Ian MacLaren, Tel: (44) (0) 121 414 3447 } IRC in Materials for FAX: (44) (0) 121 414 3441 } High Performance Applications, email: I.MacLaren-at-bham.ac.uk } The University of Birmingham, or: ianmaclaren-at-hotmail.com } Birmingham B15 2TT, http://web.bham.ac.uk/I.MacLaren } England. } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } } }
Hi Doug, I'm in almost the same situation that you describe. Can you post a summary once you have assembled replies to these questions? Here are a couple of URL's that I used when I was searching the web for info. Caution though, I got really bogged down. As you say, lots of glossy brochures but it's difficult to read between the lines (note the resolution pun) when you are limited on experience. Good luck. John
I've had good results using a Polaroid SprintScan45. They gave me a steel plate adapter with magnetic strips to insert in the 4x5 negative carrier and this worked fairly well. I went a step further and had a replacement plate made for TEM negatives that replaces the 4x5 spring loaded plate that comes with this holder. It works fairly well. The scanner is reasonably priced. Check out Polaroid's web site for specifications. -Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Ian MacLaren To: Microscopy-at-Sparc5.Microscopy.Com Cc: g.r.millward-at-BHAM.AC.UK -----------------------------------------------------------------------.
Dear all, I am passing this message on for a colleague who is wondering about purchasing a better scanner, specifically for TEM plate negatives. He already has a UMAX Astra 1200S with transparency adaptor but this does not cope with high optical densities, or with enlarging small areas greatly. We use two different sizes of plate film: one just smaller than 3 1/4" by 4" and the other 2 1/2" by 3 1/2". What would you recommend, a high end flatbed with transparency adaptor or a specialist negative scanner? Which models have worked well for you. He would prefer it to be Mac compatible.
Thanks
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, or: ianmaclaren-at-hotmail.com Birmingham B15 2TT, http://web.bham.ac.uk/I.MacLaren England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Our group currently has a JEOL FX 2000 TEM available for sale to anyone interested. The system includes a cold and hot stage, PC driven X-ray system, and STEM attachment. Please respond if interested.
M.W. Rigler, Ph.D. MAS, Inc. Suwanee GA 770-866-3218
} Nestor's caution is fully justified. His description is right on the mark, } and the rules have just been re-issued as Important Notice number 122 of } 1998 (see http://www.nsf.gov/pubs/1998/iin122/iin122.txt). } } Caroline was generous. We charge commercial customers three times our } academic rate - but the academic rate is the same for MIT and any other } academic user. } } Tony. } I've gotten similar comments offlist; apparantly I didn't make my point clearly. Two times academic rates was a university-wide policy, and I had no authority to change it (tho I wanted to). Anyone setting up a new system needs to check on the existence of similar rules.
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I am looking for a source to buy a Durst Laborator enlarger (with a point source and variable condensors) for printing our TEM negatives. I have contacted several sources on the web, including Photoshopper and Durst ACS, Inc., and have been unsuccessful in eliciting ANY responses from ANY vendor for purchasing a new Durst enlarger. I understand from my local photog supplies vendor that Durst is a quirky company with only a few authorized reps in the U.S.
Can anyone help me find a Durst vendor?
Offline replies would be fine. If others express interest, I will publish what I learn. Thanks to all.
Ann Hein Lehman TEM Lab Mgr, Trinity College LSC 314 300 Summit St. Hartford, CT 06106 860-297-4289 Ann.Lehman-at-exchange.cc.trincoll.edu
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good luck. I had a helluva time getting any info out of their Midwest representative (some camera shop in Milwaukee) several years ago. I don't see how they stay in business. However, if you do find someone who will actually talk to you, please send me their name. I still have some questions about the finer details of operation. On the other hand, maybe with you could pick up a good used enlarger from a lab going digital.
The MSA office received a request from a high school recently for a brochure on "careers in microscopy". MSA doesn't have one, but I suspect that some of you who read this list may. If you have one, will you please send me a sample so that MSA and MICRO can refer future inquiries?
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Soon after I asked the question about the ion-milling with a cooled stage, I received a great deal of information concering this problem. Now, we solve it. Many thanks for your kind suggestion and help.
Yours Sincerely,
J.S. Wu --------------------------------- Jinsong WU LSG2M, Ecole des Mines de Nancy Parc de Saurupt F-54042 NANCY cedex France
Caroline Schooley wrote: ============================================= The problem in Hawaii was different; it had to do with the way lab usage was being charged to federal grants. I dealt with commercial usage pre- retirement at U.C. Berkeley. There were very specific statewide university rules that covered all such use on the campus. I was allowed to provide service to outsiders only if there was no private lab in the local area offering the same service, and I was told to charge double the campus recharge rate. I supported the policy, because competition with private enterprise using grant-funded equipment really is unfair. =================================================== Thanks Caroline, I am sure that I can speak for the many others performing electron and light microscopy as a for-profit tax-paying laboratory service wishing that the idea of this kind of "unfairness" was more universally recognized (and appreciated).
However, more often than not, such policies are formulated in ways that, like no-fault automobile insurance in many states, sound like they are accomplishing something of significance but in the end the life goes on for the lawyers as if nothing had changed.
} From my own almost thirty years of experience with this issue, from what I have seen, one would have to multiply the internal charge rate, in many instances, by factors of 5X to 10X before the "selling price" to the client would be truly "competitive" with those of commercial firms offering comparable services.
Second, the idea of there being "no private lab in the local area offering the same service," does not work either. For one thing, how does one define "local"? And how does one determine if it is the "same service"? We recently lost what for us would have been a nice SEM job, it was a large number of repetitive samples of glass spheres, highest magnification being about 1000X, to a university that justified their being able to do the work because they had an FE-SEM and we had a more plebeian tungsten filament SEM. And of course, the client had to drive about five miles further to get to us than to the university. So those those who manage the laboratory, and who have the most to benefit either via the normal structure of rewards present in such settings or via outright consulting fees, are also the ones making the decisions as to what is "local" and what is the "same". Sort of seems like it is the fox guarding the chicken coop (at least in some instances).
NSF Important Notice 122, like with the no fault insurance laws have the appearance like it is accomplishing something but those of us on the firing line on a daily basis, having to compete in the marketplace with university competitors, know that in most instances such policies are just window dressing and don't, at the end of the day, accomplish the stated goals.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI Structure Probe, Inc. FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I have a student attempting to immunogold label G-protein in arabidopsis guard cells. The antibody is made to aribidopsis protein expressed in E. coli. We can easily label E. coli cells expressing the protein and the antibodies recognize the protein purified from plants and run on westerns. No light level work has been done. However we are having a number of problems attempting to label the protein in Arabidopsis: the ultrastructure is terrible, and we have not even a hint of label in the plants. Any advice would be appreciated.
I would also appreciate advice on how to handle Arabidopsis guard cells for EM. We are trying leaf peels and diced leaves but both are difficult to consistantly section and get good guard cells. The student attempting this project, and doing all the embedding and sectioning, is an undergraduate; so any surefire advice you can offer for sectioning guard cells would help.
Thanks in advance,
Michelle
#################################################### Michelle Peiffer Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:mlk101-at-psu.edu ####################################################
I think we need some additional information on the nature of the Arabidopsis protein and how you are doing your specimen prep before we could make any recommendations.
Since you can label it in E. coli and it shows up OK on blots, it sounds like the antibody is doing what it's supposed to do. Since I'm not a botanist, I will have to ask a basic question: what is the nature of the protein in native guard cells? Is it water soluble? Hydrophobic? A membrane protein?
You mention that the ultrastructure is terrible, and this may be the root of the problem (no botanical pun intended!). How are you fixing and embedding the specimens, and what resin are you using? Also, what about the immunolabeling procedure? Are you using Protein A-gold or a "bridge antibody" like maybe goat anti-rabbit IgG and then streptavidin-gold?
Sorry I have more questions than answers. Maybe there's someone else out there that does this kind of work on a regular basis and can give you a more direct answer. If not, maybe we can help if you provide some additional details.
Best wishes, Bob **************************************** Robert (Bob) Chiovetti Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives / Systems Integrators / Analog & Digital Imaging *****************************************
It seems to me that the last time this "thread" on NSF document 122 and outside service work went through the listserv the issue of when NSF ownership of a piece of equipment ends came up......anyone recall something along these lines? If one uses the standard of 10% depreciation/yr, then any piece of equipment housed in a University - even though NSF-bought - has minimal value at the end of 10 years. Add on increasing maintenance and service costs as the equipment ages and you can be sure that there is a "crossover point" at which the University has more invested on an annual basis than the equipment is worth. At that point, ownership comes into question.
I appreciate that this too is a "fine point", but there is ownership, and there is ownership.
Winton
PS: I have no NSF-bought equipment
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
The Electron Microscope Unit at the University of New South Wales has available for sale one JEOL WDS Spectrometer (in excellent condition - hardly used) suitable for the JEOL 840 / JXA- 8600S series of electron microscopes. Two light element crystals - STE (sterate) and TAP - thallium acid phthalate - are also available for sale. Also on offer is one set of an early 1980's vintage counting electronics/hv supply in a JEOL cabinet (~48" x 24" x 24"). The EM UNIT is seeking expressions of interest on any one or all of the above items. Please respond to me by Email if you are interested. Thankyou Barry EM UNIT UNSW
For Ren-Jye and others interested in preparing in particular: stained sections of polymers for TEM, and in general: all kinds of materials ranging from composite to catalysts;
may I recommend the following review:
H.K.Plummer - "Reflections on the use of microtomy for materials science preparation"
Microsc. Microanal. 3, 239-260 (1997)
Which draws on years of experience at the Ford Research Laboratory.
We ourselves are etchers, and only occasionally stainers.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
HELP US! We are interessed in the acquisition of a "second hand" (old fashioned)
SEM and TEM electronic microscopes. Free of charge is also accepted and appreciated. If you think you might help us in this matter, non hesitate give me a call:
Oncological Institute BUCHAREST-ROMANIA Corneliu Mateescu Ph.D cmateescu-at-mail.iob.ro =====================================================
I think it is important to consider the customer in this debate. We are in a slightly different situation here but have been involved in supplying a commercial service for many years. When we started providing this service we first calculated the true cost of the service. This cost incorporated all costs such as equipment price, depreciation, servicing, and operator time. Once this true cost is charged I think competition with outside services is then acceptable. Universities can provide a different element to a service which an outside service with a narrow focus cannot supply. This may be knowledge/expertise or new techniques. Is it right to deprive customers of these benefits simply because we are a University ? We have an excellent relationship with competing commercial services because we are not seen to exploit our position to undercut them. In fact our closest competitor ( six miles ) in the outside world sends work into us when they are under pressure. I should point out too that the last SEM bought for us was 17 years ago and we have funded all purchases of equipment from earnings since then. I know that our customers do not come to us because of price but because we provide a necessary service to them. Once a true commercial rate is charged I feel that the playing field is level and Universities should not be prevented from providing this service.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
-----Original Message----- } From: Garber, Charles A. {cgarber-at-2spi.com} To: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
We are looking to purchase a used EPMA, preferably a JEOL 733 or later.
Anybody got one languishing in the spare room?
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
The responses we received for the FREE SEM were overwhelming. In fact, there was a gentleman knocking at the door Monday morning and he is the lucky winner. If there is a change in plans, I will revert back to the list of inquirees and pick another winner.
} Hello! } } I am a TV researcher based in Vancouver, working on developing a TV } series about new initiatives in Microscopy. I am looking to find the } institution (s) and/or individual (s) doing the newest intiatives in the areas of: } } *Nanotechnology; } *Microscopy: techniques, technologies, Microvision, etc. } *Microelectronics; } *Microbiology: Digital Centre for Microbial Ecology, primordial } microbes? microbes on Mars, DNA microarrays, microbia physiology, } American Type Culture Collectin, microshells - shells less than 5 mm in size and microfossils; } *Microoptics } *Minerology: micrographs of rocks, crystals, etc. } *Micrometeorology } *Micromechanics: micromaching - Sensors, Actuators } *Microcirculation } } Please let me be more specific. Some possible topics, drawn from a wide range of scientific disciplines, might include: microscopy - technology, human parasites, insect flight, insect smells, nanotechnology, optical computing, fusion power, biological computers, targetted drug delivery, ocean microorganisms, the soup of life, cell regeneration, etc. } } I would appreciate any information. Since I'm looking for the newest, } not necessarily documented, initiatives in these areas - it is a little } difficult. } } PLEASE EMAIL ME DIRECTLY AT callain-at-intouch.bc.ca
I will be signing off soon - politics and early retirement strike again! Clearing my office! I want to thank you for your presence in my life in recent years. Also for responding to my questions - I really appreciate it. You are a great bunch of people. There is nothing like helpful support when you need it! I may be back in another guise. In the meantime, thanks, farewell and may the electrons be with you!
Keith Ryan Plymouth Marine Lab., UK PS - Daniele - don't cry! The e-mail address will still work, I hope. We'll meet again (now everybody wonders, who is this Daniele?!) XXX
Can you please send one to us. I have spend some time helping people with deciding on what they need. But this did not take off as I would have wished for. A opinion from a independent individual will help.
} One can make up a "decision tree" to help with this. If people are } interested, I can write up an article that walks through these steps to help } one decide on the most appropriate technology based on a set of needs. } -Ted Inoue } Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
Dear all please pass this to any relevant people. Please note imminent closing date.
RESEARCH ASSOCIATE/RESEARCH FELLOW (REF: A78/98)
CENTRE FOR MICROSCOPY AND MICROANALYSIS
The Centre for Microscopy and Microanalysis is the premier Microscopy Centre in Western Australia. In mid 1998 it was awarded a Western Australian Government Centre of Excellence in association with Curtin, Murdoch and Edith Cowan Universities.
The appointee will be required to carry out research in advanced microscopy. It is expected that the appointee will also take a leading role in the development of confocal microscopy, training and support of users, development of specimen preparation techniques, development of associated computer facilities and the support of research and teaching involving this instrument within the Centre. The minimum qualification required is a PhD in Science, Medicine or Agriculture together with experience in light and electron microscopy. Preference will be given to applicants with a substantial knowledge and skill in confocal microscopy and computing. The successful applicant must have strong interpersonal skills and the ability to work as part of the highly motivated group of academics at the Centre. The position is tenurable. For further information and copies of the selection criteria please contact either Associate Professor John Kuo on telephone (618 - international) OR (08) 9380 2765 or email jjskuo-at-cyllene.uwa.edu.au or Dr Gregory Pooley on telephone (618 - international) OR (08) 9380 2261 or email gdp-at-cyllene.uwa.edu.au or access the web link below.
SALARY RANGE: Level A $32,585 - $44,221 p.a. (Minimum starting salary for appointee with PhD will be $41,196 pa) Level B $47,946 - $56,937 p.a.
CLOSING DATE: 4 December 1998
Conditions of appointment will be specified in any offer of appointment which may be made as a result of this advertisement.
Written applications quoting reference number, telephone number, qualifications and experience and the names, addresses (including Email) and fax/telephone numbers of 3 referees should reach the Director, Human Resources, The University of Western Australia, Nedlands WA 6907, by the closing date.
http://jobs.uwa.edu.au/
The University is an equal opportunity employer and promotes a smoke-free work environment.
Brendan J. Griffin Centre for Microscopy and Microanalysis The University of Western Australia Nedlands, WA, AUSTRALIA 6907 ph 61-8-9380-2739 fax 61-8-9380-1087
Sorry that I didn't provide all the details on the original post, here they are:
We are attempting to label g-protein in Arabidopsis guard cells. Westerns indicate the protein is water soluble, but also associates with membranes, though it is not an integral membrane protein. Conventional fixation of the cells, with aldehydes, osmium, embedded in Spurrs yeilds good ultrastructure, but no labelling. For labelling we are fixing with 4% paraformaldehyde, 0.5% glutaraldehyde in 0.1 M phosphate buffer; dehydration in ethanol, embedding in LR White. The primary antibody is polyclonal, affinity purified IgG, the secondary is goat anti-rabbit conjugated to 10 nm colloidal gold. We are getting acceptable ultrastructure and labelling in E. coli (expressing the protein) processed this way, but not even a hint of labelling in plants.
We do have the resources to do cryo-work, but the equipment is all brand new, and we are still working out the details. Any suggestions on improving this protocol will be greatly appreciated. Thanks
#################################################### Michelle Peiffer Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:mlk101-at-psu.edu ####################################################
We have a Nikon TMD inverted epifluorescence microscope and a Nikon Labophot upright scope. We do not have an oil immersion lens on the inverted. Does anyone know if it would be a bad idea to take the oil immersion lens (Nikon Plan 100 (serial number 190370)) off of the upright scope and use it on the inverted? Can an oil immersion lens for an upright scope be used in the inverted position without harm?
TIA
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
Listers, A grad student in my TEM lab is requesting information regarding "calcium mapping." Her research involves calcium signalling during the metamorphosis of certain hydroids. She's using potassium pyroantimonate to precipitate free calcium during fixation/embedding. Precitate has been found in "unusual" places and calcium mapping has been suggested to verify that it is indeed calcium. Since "graduate student" is synonymous with "poverty" (her words) is there anyone out there who would offer free advice or low-cost mapping services. Please o' please (her words). Any response would be greatly appreciated. Contact can be made through me. Danke.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supervisor 11/11/98 11:12:34 AM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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I am using glycol methacrylate embedding and immunostaining to investigate polarity in small aggregates (100 microns) of hepatocytes. I have embedded the aggregates along with liver tissue and intestinal tissue (as controls) in JB4 glycol methacrylate following manufacturer's instructions. All attempts at staining so far have yielded results in the liver tissue only. No staining has been observed in the aggregates or the intestine which are significantly smaller in volume. I have been using trypsinization and several forms of "etching" I found in the literature (NaOH in 50% Ethanol/water, acetone). I am using mouse primary antibodies and goat-anti-mouse Oregon Green 488 as a secondary.
Any suggestions for getting results in the smaller tissues?
Thanks, Susan A. Fugett
Department of Chemical Engineering and Materials Science Phone: 612-625-8803 University of Minnesota 612-625-0808 421 Washington Ave SE Fax: 612-626-7246 Minneapolis, MN 55455 Email: fugett-at-cems.umn.edu
In a message dated 98-11-11 11:22:36 EST, wise-at-vaxa.cis.uwosh.edu-at-sparc5.microscopy.com writes:
{ { Does anyone know if it would be a bad idea to take the oil immersion lens (Nikon Plan 100 (serial number 190370)) off of the upright scope and use it on the inverted? Can an oil immersion lens for an upright scope be used in the inverted position without harm? } } Hi Bob,
You can certainly do this, it won't hurt a thing, in fact one of my customers has set up just such a situation on a Nikon inverted scope. But keep a couple of things in mind:
1. Use the bare minimum of oil that is necessary to fill the gap. Oil will tend to run down the lens, and this may cause a bit of a mess. Have lots of lens paper handy!
2. The oil lens probably has a fairly short working distance. This means that you may not be able to focus through a microscope slide or a culture plate, for example. But there are some options here, as well:
2A. If your specimen has to be on a microscope slide, you could seal the coverslip with paraffin wax or Vaseline and invert the whole slide, I suppose.
2B. If the specimen has to remain upright, you could place it on a thin coverslip and make a holder for the stage with an open space for the coverslip. There should be enough working distance in the lens to focus on the far (upper) surface of the coverslip.
3. If the specimen is still out of the focal range of the lens, you may have to modify the stage or make a simple flat metal plate that would substitute for the stage on the scope. My customer had to take this route for his setup, but it works just fine.
Hope this helps.
Cheers, Bob **************************************** Robert (Bob) Chiovetti Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems *****************************************
Bob: We use our inverted Nikon with oil immersion lenses every day for the last 5 years with no trouble. Use a minimum amount of oil, clean well when you are done. There have been reports on the listserver (either microscopy or confocal) where users have gotten oil into the innards of the objective and some users have made dams out of various things (e.g., rubber o-rings) but we have avoided that. A NA 1.4 condenser on an upright scope is, of course, meant to be oiled in the same way and they generally survive (tho an MD ruined the condenser in one of my old labs by not cleaning up after himself when he was done). Good luck. Tom
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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In a message dated 98-11-11 11:22:36 EST, wise-at-vaxa.cis.uwosh.edu-at-sparc5.microscopy.com writes:
{ { Can an oil immersion lens for an upright scope be used in the inverted position without harm? } } Bob,
One thing I forgot to mention: Several scope manufacturers (I don't know about Nikon) use a standard lens thread on their upright scopes and an "RMS" thread on their inverted scopes. If this is the case you will need an adapter collar which screws on the lens to convert it to an RMS thread for the inverted scope. I know Leica has such collars, so surely Nikon also has them if they are needed.
Keep in mind that using a collar on the lens *may* cause a slight parfocality problem.
Try gently screwing the lens into the inverted scope's nosepiece turret. If the lens does not mate properly, you will need an adapter collar.
Bob **************************************** Robert (Bob) Chiovetti Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging *****************************************
Thanks all - 23 responses to this posting (so far). I will summarize/respond in turn ASAP.
-----Original Message----- } From: Lehman, Ann Sent: Monday, November 09, 1998 10:37 AM To: 'MSA Listserver'
Winston Wiggins wrote: } } Listers, } A grad student in my TEM lab is requesting information } regarding "calcium mapping." Her research involves calcium signalling } during the metamorphosis of certain hydroids. She's using potassium } pyroantimonate to precipitate free calcium during fixation/embedding. } Precitate has been found in "unusual" places and calcium mapping has } been suggested to verify that it is indeed calcium.
What concentration of Ca is she looking for? The usual con- centrations involved in signalling are lower than micromolar, and are exceedingly difficult to impossible to determine with microanalytical techniques. The Somlyos have succeeded in seeing very low Ca concen- trations with EELS, and Marie Cantino reported using x-ray microanaly- sis to determine Ca (Proc MSA/MAS 1998) also at low levels. WDS could conceivably have sufficient sensitivity, but I don't know whether any- one has used it for these kinds of studies.
} Since "graduate student" is synonymous with "poverty" (her words) } is there anyone out there who would offer free advice or low-cost mapping } services. Please o' please (her words). Any response would be greatly } appreciated. Contact can be made through me.
Our facility has EDS (which can detect millimolar [Ca] or so, depending on the matrix), and we are a biotechnological resource, so we would be free (except for travel, hoterl, meals, etc.), but we are not likely to be suitable unless [Ca] is fairly high. Good luck. Yours, Bill Tivol
One of the vacuum pumps servicing our electron microprobe just gave up the ghost. We are faced with replacing the pump as this particlur pump from Edwards (an EDM-12) is obsolete - thus, rebuild kits are unavailable from Edwards. (if rebuilding were the way we wanted to go)
Edwards has a stock of a rebuilt pump that is a successor to ours (2 generations removed), with these rebuilt to factory specs. (this pump is the RV-12). I can get one for a reasonable price, which is about 60% of the cost of a new pump.
My questions to you:
1. how do you feel about the "vacuum quality" of rebuilt pumps vs. new pumps
2. how close do I have to get in pump specs?....the new pump has a capacity of 17.0 m^3/hr, while the old has (had) a capacity of 17.5 m^3/hr, i.e., there is about a 3% difference between them......should I shoot for a higher pumping capacity relative to the the old pumps?
Thanks, in advance, for your responses.
Winton
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
So a little while ago I asked about prep'ing suspension cells for SEM. I got a lot of helpful suggestions which I will summarize & post here when I get the time. It ends up being that the guy grew them on gelatin coated coverslips and chamber slides which are WAY too big for my cpd. So I'm either going to have to try to cut or break them to make them fit. Or I was considering HMDS, has anybody out there used HMDS for things as delicate as cells? If you have could you e-mail me with protocols? I have the one that works for bugs and I have the HMDS, I've just never used it. Any help you send my way will be greatly appreciated. I'm getting sooo smart from y'all's ideas that my head hurts.
Going quietly into the SEM room,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Try etching for 3-5 minutes in sodium ethoxide soln 1:1 with toluene, = then go to 100% EtOH down to water, buffer, etc. Ethoxide Solution: 75gm NaOH in absolute ethanol; let stand 10-14 days or until pellets = dissolve with occasional stirring. Solution should turn brown when = ready. This works well with tissue embedded in Araldite 502 (and may = work with JB4 media) allowing all types of histochemistry procedures and = immunohistochemistry.
Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
-----Original Message----- } From: Susan Fugett [SMTP:fugett-at-cems.umn.edu] Sent: Wednesday, November 11, 1998 5:26 PM To: microscopy listserver
I am using glycol methacrylate embedding and immunostaining to=20 investigate polarity in small aggregates (100 microns) of hepatocytes. I =
have embedded the aggregates along with liver tissue and intestinal=20 tissue (as controls) in JB4 glycol methacrylate following manufacturer's =
instructions. All attempts at staining so far have yielded results in = the=20 liver tissue only. No staining has been observed in the aggregates or = the=20 intestine which are significantly smaller in volume. I have been using=20 trypsinization and several forms of "etching" I found in the literature=20 (NaOH in 50% Ethanol/water, acetone). I am using mouse primary = antibodies=20 and goat-anti-mouse Oregon Green 488 as a secondary.
Any suggestions for getting results in the smaller tissues?
Thanks, Susan A. Fugett
Department of Chemical Engineering and Materials Science Phone: 612-625-8803 University of Minnesota 612-625-0808 421 Washington Ave SE Fax: 612-626-7246 Minneapolis, MN 55455 Email: fugett-at-cems.umn.edu
This is a multi-part message in MIME format. --------------0ACCEBDC64F1D9ABE2ADF72B Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
"Vacuum quality" which really means base pressure and backstreaming is more effected by the choice of pump oil than mechanical matters. If the rebuild was done properly there is no problem.
Pumping speed is just that. The new pump will be 3% slower to pump down. The length of vacuum hoses and the presence of foreline traps effects pumping speed more. The only slow pumping speed problem I have seen was where a new microscope was installed with a trap and too long of a vacuum line so that the evacuation control electonics and valves went into oscillation at crossover because of slow pumping and some outgassing.
Ronald Vane XEI Scientific 650-369-0133
Winton Cornell wrote:
} Folks: } } One of the vacuum pumps servicing our electron microprobe just gave up the } ghost. We are faced with replacing the pump as this particlur pump from } Edwards (an EDM-12) is obsolete - thus, rebuild kits are unavailable from } Edwards. (if rebuilding were the way we wanted to go) } } Edwards has a stock of a rebuilt pump that is a successor to ours (2 } generations removed), with these rebuilt to factory specs. (this pump is } the RV-12). I can get one for a reasonable price, which is about 60% of the } cost of a new pump. } } My questions to you: } } 1. how do you feel about the "vacuum quality" of rebuilt pumps vs. new pumps } } 2. how close do I have to get in pump specs?....the new pump has a capacity } of 17.0 m^3/hr, while the old has (had) a capacity of 17.5 m^3/hr, i.e., } there is about a 3% difference between them......should I shoot for a } higher pumping capacity relative to the the old pumps? } } Thanks, in advance, for your responses. } } Winton } } Dr. Winton Cornell } Senior Research Associate & Supervisor, Microanalysis Laboratory } Department of Geosciences } The University of Tulsa } 600 South College } Tulsa, OK 74104-3189 } } phone: 918-631-3248 } fax: 918-631-2091 } e-mail: wcornell-at-centum.utulsa.edu
} Or I was considering HMDS, has anybody out there used HMDS for } things as delicate as cells?
Shirley Pinchuck, in this lab, has done a fair amount of work using HMDS, including comparisons of HMDS with other methods such as CPD, cryo-SEM, etc. I will ask her to fax you her protocols as well as an abstract of a conference presentation on some of the comparative work.
I hope this helps.
Regards
Robin
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
There should not be any difference in performance between a new pump and a factory rebuilt one - they should both acheive the original vacuum specification. Obviously the rebuilt one may not look as shiny but we have been happy with rebuilt pumps over the years.
I assume that the pump is backing a high vacuum pump of some sort in which case it has to meet the specification for that, 3% is not significant. If your microprobe uses a Diffstack 160 (guessing at 1 vacuum supplier) then the RV12 is the currently reccomended backing pump.
Regards, Ron
} Folks: } } One of the vacuum pumps servicing our electron microprobe just gave up the } ghost. We are faced with replacing the pump as this particlur pump from } Edwards (an EDM-12) is obsolete - thus, rebuild kits are unavailable from } Edwards. (if rebuilding were the way we wanted to go) } } Edwards has a stock of a rebuilt pump that is a successor to ours (2 } generations removed), with these rebuilt to factory specs. (this pump is } the RV-12). I can get one for a reasonable price, which is about 60% of the } cost of a new pump. } } My questions to you: } } 1. how do you feel about the "vacuum quality" of rebuilt pumps vs. new pumps } } 2. how close do I have to get in pump specs?....the new pump has a capacity } of 17.0 m^3/hr, while the old has (had) a capacity of 17.5 m^3/hr, i.e., } there is about a 3% difference between them......should I shoot for a } higher pumping capacity relative to the the old pumps? } } Thanks, in advance, for your responses. } } Winton } } } Dr. Winton Cornell } Senior Research Associate & Supervisor, Microanalysis Laboratory } Department of Geosciences } The University of Tulsa } 600 South College } Tulsa, OK 74104-3189 } } phone: 918-631-3248 } fax: 918-631-2091 } e-mail: wcornell-at-centum.utulsa.edu } } } }
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Our group currently has a JEOL FX 2000 TEM available for sale to anyone interested. The system includes a cold and hot stage, PC driven X-ray system,and STEM attachment. Please respond if interested.
M.W. Rigler, Ph.D. MAS, Inc. Suwanee GA 770-866-3218
Hi group, I am posting this message to resume the problem that was posted some time ago (17.09.98 ' latex microsheres') with request on info about particle suppliers. I would like to thank everybody who replied but the problem was that NO ONE company from the list we received had right particles(1-10 nm, optically transparent at a given wavelength region and absorbing at different given region). Finally I have found the best source for not only me as I guess : SPHEROTECH INC, 847-680-8922, http://www.spherotech.com with the widest range of particles for microscopy needs. Best regards Dmitri Lapotko Luikov Heat and Mass Transfer Insitute Minsk Belarus
Bob, Having been a dealer rep for Nikon, I respond this way. You can invert the lens and use it without adapters or problems with parfocality, however, the working distance is very limited and would only work when the sample is resolved through a cover glass. This tells you that the sample could just as easily be viewed on a compound microscope. Nikon does offer long working distance lenses specifically designed for the TMD. I do not recall a 100X oil ever being offered, but they do offer a 60X ELWD dry.
I am teaching histology and microtechnique to advanced high school students. I need a source of good prepared demonstration slides. Somewhere along the way I lost my collection which I made when I worked in a medical lab. Because I am paying for the slides myself they need to be economical. I bought some from Edmund Scientific and found that they were unuseable, pathetic junk. Can anyone suggest good quality sources for prepared slides that I might be able to afford. I am especially interested in series of slides of the same specimen that use a variety of stains. I would also be interested in purchasing slides from any of the experts on this mailing list who must have great examples of their skills which would be appropriate for educational purposes.
It is possible that the statement below about the speed of the pump being 3% slower may not be relevant to your situation. What should be taken into consideration in selecting a pump is the effective pump speed. The effective pump speed is the speed at the vacuum chamber port when the capacitance of the tubing from the pump to the vacuum system is taken account. It works like parallel resistors. The effective pump speed is given as
where S(pump) is the pump speed and C(tubing) is the capacitance of the tubing. A system can easily become capacitance limited if C(tubing) { { S(pump). When this happens, Seff=S(pump). If this is the case, the 3% difference in pump speeds will not matter. Standard books on vacuum science can give you the values of capacitance for the length and diameter of tubing, elbows, valves, and how they are added to give and effective value to the pump.
So, the moral of this story is that you should always consider your total vacuum system when selecting a pump.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Ronald Vane To: Winton Cornell Cc: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Winton and all:
The rebuilt pump should be just fine. Go for it.
"Vacuum quality" which really means base pressure and backstreaming is more effected by the choice of pump oil than mechanical matters. If the rebuild was done properly there is no problem.
Pumping speed is just that. The new pump will be 3% slower to pump down. The length of vacuum hoses and the presence of foreline traps effects pumping speed more. The only slow pumping speed problem I have seen was where a new microscope was installed with a trap and too long of a vacuum line so that the evacuation control electonics and valves went into oscillation at crossover because of slow pumping and some outgassing.
Ronald Vane XEI Scientific 650-369-0133
Winton Cornell wrote:
} Folks: } } One of the vacuum pumps servicing our electron microprobe just gave up the } ghost. We are faced with replacing the pump as this particlur pump from } Edwards (an EDM-12) is obsolete - thus, rebuild kits are unavailable from } Edwards. (if rebuilding were the way we wanted to go) } } Edwards has a stock of a rebuilt pump that is a successor to ours (2 } generations removed), with these rebuilt to factory specs. (this pump is } the RV-12). I can get one for a reasonable price, which is about 60% of the } cost of a new pump. } } My questions to you: } } 1. how do you feel about the "vacuum quality" of rebuilt pumps vs. new pumps } } 2. how close do I have to get in pump specs?....the new pump has a capacity } of 17.0 m^3/hr, while the old has (had) a capacity of 17.5 m^3/hr, i.e., } there is about a 3% difference between them......should I shoot for a } higher pumping capacity relative to the the old pumps? } } Thanks, in advance, for your responses. } } Winton } } Dr. Winton Cornell } Senior Research Associate & Supervisor, Microanalysis Laboratory } Department of Geosciences } The University of Tulsa } 600 South College } Tulsa, OK 74104-3189 } } phone: 918-631-3248 } fax: 918-631-2091 } e-mail: wcornell-at-centum.utulsa.edu
HMDS can work fine for cells. You might have to play a bit to find the best transition series, but I'd start with 3:1-} 1:1-} 1:3 absEtOH:HMDS. Drying is the bigger question: how fast at what temperature? If I may, we ran an article on this in the May '97 issue of Microscopy Today. If you don't have a copy, I can see if we've got an extra one.
Information on HMDS has also been posted on the U Florida Tips & Tricks of Microscopy pages: http://www.biotech.ufl.edu/~emcl/tips.html
I can send more details if you wish.
Phil
} It's me again, } } So a little while ago I asked about prep'ing suspension cells for } SEM. I got a lot of helpful suggestions which I will summarize & post here } when I get the time. It ends up being that the guy grew them on gelatin } coated coverslips and chamber slides which are WAY too big for my cpd. So } I'm either going to have to try to cut or break them to make them fit. } Or I was considering HMDS, has anybody out there used HMDS for } things as delicate as cells? If you have could you e-mail me with } protocols? I have the one that works for bugs and I have the HMDS, I've } just never used it. } Any help you send my way will be greatly appreciated. I'm getting } sooo smart from y'all's ideas that my head hurts. } } } Going quietly into the SEM room, } } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } phone: 510-642-2085 } fax: 510-643-6207 } http://biology.berkeley.edu/EML
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net
This is the promised follow up on my question (posted 11/6/98) about color CCD cameras, included is my original posting and the responses I received to date. I really appreciated the "low key" and informative responses of some of the vendors.
I should add that I found some of the information that helped me frame my questions from this site: http://www.soundvisioninc.com/howdcw.htm
Doug Cromey
--------------the original question---------------------
} Microscopy Listers, } } We are in the early stages of evaluating a digital imaging system here (for } light microscopy). I'm trying to understand the different ways that CCD } cameras can be or are used to acquire color images. There seem to be } several ways this is done: } } * Single chip Monochrome CCD array with some type of filter in } front of the camera to allow it to acquire sequential red, } green & blue images and then software to "reassemble" the } images into a color image. } * 3 Chip camera, with each chip assigned (filtered for) red, } green & blue and then software to "reassemble" the images into } a color image. } * Single chip CCD with some type of color mosaic "mask" on the } chip to acquire the red, green & blue parts of the image and } then software to "reassemble" the images into a color image. } * CCD array where the image is "scanned" either by moving optical } elements or moving the CCD array to acquire a high pixel count } with a fairly small sensor. These would be the slowest for } acquisition, but I gather they give a lot of "bang for the } buck (euro)". } * Others? } } What are the Pros & Cons of these different types of cameras? Is there a } WWW or published resource that could help me sort this out? } } Our interest here is primarily in acquiring static images (not real-time } video) from low light level fluorescence, DIC and/or bright field. We } would like the camera to be sensitive enough for quantitation if needed. } I'd also like to be able to advise others here who may have different } requirements. I'm not specifically "fishing" for sales pitches (I already } have plenty of glossy literature, I'm just trying to make sense out of it). } } I would be happy to take replys "off-list" and post a summary. } } Yours, } Doug Cromey
----------reply--------------
} From: RCHIOVETTI-at-aol.com
good for resolution and, if properly used, color fidelity but not good for motion or dynamic or time sensitive subjects (like fading flourescence)
} * 3 Chip camera, with each chip assigned (filtered for) red, } green & blue and then software to "reassemble" the images into } a color image.
reasonably good color fidelity but the dichroic filters suck up lots of the light and require special care with the selection of the adapter - dichroics what parallel light or you'll get funny color fringes and shifts
} * Single chip CCD with some type of color mosaic "mask" on the } chip to acquire the red, green & blue parts of the image and } then software to "reassemble" the images into a color image.
color fidelity is not as good - most have 1/2 green, 1/4 blue and 1/4 red but at least one (Panosonic GP-KS1000) has more pixels (900,000 vs 410,000) and a different color array of 1/3 each of R/G/B
} * CCD array where the image is "scanned" either by moving optical } elements or moving the CCD array to acquire a high pixel count } with a fairly small sensor. These would be the slowest for } acquisition, but I gather they give a lot of "bang for the } buck (euro)".
scanned arrays can give unbelivable resolution - I just saw a one this weeek that used a 10K Kodak linear array! but the time to scan is often the limiting factor with all that that implies (see - http://betterlight.com)
} * Others?
CMOS - new technology, less expensive (see http://www.soundvisioninc.com) Digital cameras - if you never need "video" these can be your salvation, just decide what you need to do and find one that does it (see http://www.electrim.com or the guy from DVC ) there are also CID cameras (low blooming)
There are also a host of all of the above with low light capabilities either through such technology back thining or on chip integration
} } What are the Pros & Cons of these different types of cameras? Is there a } WWW or published resource that could help me sort this out? } } Our interest here is primarily in acquiring static images (not real-time } video) from low light level fluorescence, DIC and/or bright field. We } would like the camera to be sensitive enough for quantitation if needed. } I'd also like to be able to advise others here who may have different } requirements. I'm not specifically "fishing" for sales pitches (I already } have plenty of glossy literature, I'm just trying to make sense out of it). } } I would be happy to take replys "off-list" and post a summary. } } Yours, } Doug } .................................................................... } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : } : Research Specialist, Principal University of Arizona : } : (office: AHSC 4212A) P.O. Box 245044 : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : } : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): } :...................................................................: } http://www.pharmacy.arizona.edu/exp_path.html } Home of: "Microscopy and Imaging Resources on the WWW"
BTW - I don't sell any of this stuff.
---------------------reply-----------------------
} From: Thomas A Baginski {tombg-at-bictom.usuf1.usuhs.mil}
Dear Australians,
I had already deleted the message, when I found myself in contact with someone with experience in confocal microscopy who might be interested in the Confocal job. Could you please re-send the advert to me?
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I am working with a mixture of organic and water on observation using optical microscope. Here is no contrast between water and organic. I need to know if the "droplets" observed are water droplets, or organic. Is there a die or similar available to colour water so that there is a contrast between water and organic?
Zhaoxia Zhou Department of Materials Engineering Brunel University, UK Tel: +44-1895-274000-2968(o) E-mail: Zhaoxia.zhou-at-brunel.ac.uk
Could something be sent to this list too please - it would certainly interest me.
thanks
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: ROBIN CROSS To: Paula Sicurello Cc: microscopy
Hello Paula
} Or I was considering HMDS, has anybody out there used HMDS for } things as delicate as cells?
Shirley Pinchuck, in this lab, has done a fair amount of work using HMDS, including comparisons of HMDS with other methods such as CPD, cryo-SEM, etc. I will ask her to fax you her protocols as well as an abstract of a conference presentation on some of the comparative work.
I hope this helps.
Regards
Robin
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
by newton.wadsworth.org (8.8.8/8.8.8) with SMTP id NAA29824; Thu, 12 Nov 1998 13:06:42 -0500 (EST) Sender: tivol-at-wadsworth.org Message-ID: {364B210D.41C6-at-wadsworth.org}
Zhaoxia Zhou wrote: } } I am working with a mixture of organic and water on observation using } optical microscope. Here is no contrast between water and organic. I need } to know if the "droplets" observed are water droplets, or organic. Is there } a die or similar available to colour water so that there is a contrast } between water and organic? } Dear Zhaoxia, Colored ions could solve your problem (depending on whether there is significant partitioning into the organic). There is also a potential problem, since the ions might interfere with whatever else is happening. Cu++ is a nice blue color, and the intensity is greatly enhanced in the presence of NH4+. If you can add CuSO4 and NH4OH to the water without any problem, then you should see contrtast. However, if the organic has the possibility of forming N ligands, e.g., phenanthroline, the Cu could also color the organic. Also, if the organic has COOH groups with high pK's then the NH4OH could cause mixing of the organic and H2O due to the COO- groups formed at high pH. Good luck. Yours, Bill Tivol
I was wanting to know if anyone has used any wax or etch resist which will protect against warm KOH. I need to mask a Si/SiO2/Si wafer (Silicon on Insulator) to preferentially remove a the thick Si backside of my specimen and leave the surface Si/SiO2 bilayer intact for TEM imaging. I was going to use warm KOH (50 C) to remove the Si but I need to mask the surface and define a window on the backside Si to etch through (the KOH will stop at the SiO2 layer). Any suggestions?
John --------------------------------------- John Michael Glasko Materials Science and Engineering Dept. 2142 Burlington Labs/Yarborough St Raleigh, NC 27695-7916 USA tel: (919)-515-7217, fax: (919)-513-1699 jmglasko-at-unity.ncsu.edu
I've never prepared a Schiff reagent before and I'm having problems. I've been using the protocol in Presnell and Schreibman's edition of Humason's Animal Tissue Techniques, which recommends: *** basic fuchsin 0.5-1.0 g water 85 ml sodium metabisulfite 1.9 g 1.0 N HCl 15 ml
Shake for 2 hr or let sit overnight. Add 2 g of activated carbon, shake for about a minute, then filter. The solution should be water-clear. ***
Well, my solution is not water clear. In my hands, the dye never dissolves completely, and the activated carbon has absolutely no effect. P/S say that if the solution doesn't clear the charcoal is old, but I've been using a fresh batch. The resulting filtered solution is pale orange.
Any suggestions?
Gary Radice 804-289-8107 Department of Biology 804-289-8233 (FAX) University of Richmond gradice-at-richmond.edu Richmond VA 23173
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I have been asked to find out if anyone can share any information about silicon nitride or silicon oxide supports for TEM. We are interested in 100 nm thick plates or sheets - where to get them or how to make them; and on my part, how they are used.
Thanks in advance,
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-med.mcgill.ca
Gary, that is not an uncommon problem. It depends on your source of = basic fucshin which is if my memory serves me right is a combination of = pararosaline dyes. Some sources are better than others and I went thru = quite a few. I had absolutely perfect results with some produced from = Sigma under pararosaniline in their catalogue, though I don't remember = which salt I used. The solution came out perfectly clear and I stored = aliquots at minus 20C and it seemed quite stable. =20
Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
Gary, It has been awhile since I worked with the Schiff reagent but I used to mix up large batches of it. Firstly, the formula that I have (Lillie (1954) is slightly different:
basic fuchsin (C.C.) 1 gm (best to use a dye certified by the Biological Stain Commission) distilled water 80 ml NaHSO3 is 2 gm or 1.9 gm Na2S2O5 N HCl 20 ml
I noticed that this formula uses more bisulfite and more HCl -- both of which generate more SO2 for decolorization of the fuchsin.
This should go in a tightly stoppered container (to keep in the SO2 which does the decolorization of the fuchsin) and shaken every 10 minutes for a period of 2 hr. Add 500 mg finely powdered fresh charcoal (we used coconut charcoal). Charcoal must be finely ground (like talc). You can do this twice to remove any residual color. Sometimes I even added charcoal, shake and store in a refrigerator overnight. Filter through several layers of paper filter media. When colorless store at 5 deg C. If white precipitate shows up, it can be refiltered. Pink color means time to discard it.
Since your stain is not dissolving, something may be wrong with the fuchsin. Lack of destaining may be caused by not enough bisulfite or loose fitting container. Most often, we did not even need to use the charcoal.
} I've never prepared a Schiff reagent before and I'm having problems. I've } been using the protocol in Presnell and Schreibman's edition of Humason's } Animal Tissue Techniques, which recommends: } *** } basic fuchsin 0.5-1.0 g } water 85 ml } sodium metabisulfite 1.9 g } 1.0 N HCl 15 ml } } Shake for 2 hr or let sit overnight. Add 2 g of activated carbon, shake for } about a minute, then filter. The solution should be water-clear. } *** } } Well, my solution is not water clear. In my hands, the dye never dissolves } completely, and the activated carbon has absolutely no effect. P/S say that } if the solution doesn't clear the charcoal is old, but I've been using a } fresh batch. The resulting filtered solution is pale orange. } } Any suggestions? } } Gary Radice 804-289-8107 } Department of Biology 804-289-8233 (FAX) } University of Richmond gradice-at-richmond.edu } Richmond VA 23173
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
It's been a while since I made my own Schiff's but I remember the frustration of the filtrate coming out colored (pink or orange) even after repeated shaking with activated charcoal. The protocol that I used was different from the one you describe (but close I think) so I hope this helps. The problem was that the flasks I was filtering into weren't completely dry, i.e. I would filter into them, rinse them and then filter into them again thinking "this is OK because the Schiff's solution is mostly water anyway." Water is an aldehyde and reacts with Schiff's to form a pink color. By ensuring that glassware is completely dry you should achieve a colorless Schiff's solution after a couple of charcoal treatments. Water in the atmosphere may be responsible for the sneaky way Schiff reagent has of turning from an insignificant appearing clear spill into an angry red stain on hands, clothes and benchtops. This is a good indicator of how sloppy one is being in the lab. Get the stuff on your hands a couple of times and you'll learn to be careful. BTW, this sort of trouble with home-brew Schiff's has inspired many people to buy the stuff ready made. Good luck, John } ----------------------------------------------------------- } } I've never prepared a Schiff reagent before and I'm having problems. I've } been using the protocol in Presnell and Schreibman's edition of Humason's } Animal Tissue Techniques, which recommends: } *** } basic fuchsin 0.5-1.0 g } water 85 ml } sodium metabisulfite 1.9 g } 1.0 N HCl 15 ml } } Shake for 2 hr or let sit overnight. Add 2 g of activated carbon, shake for } about a minute, then filter. The solution should be water-clear. } *** } } Well, my solution is not water clear. In my hands, the dye never dissolves } completely, and the activated carbon has absolutely no effect. P/S say that } if the solution doesn't clear the charcoal is old, but I've been using a } fresh batch. The resulting filtered solution is pale orange. } } Any suggestions? } } Gary Radice 804-289-8107 } Department of Biology 804-289-8233 (FAX) } University of Richmond gradice-at-richmond.edu } Richmond VA 23173
________________________ C. John Runions, Ph.D. Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
I have gotten some pretty good TEM's of a negative stained bacteriophage prep but, unlike some published photos of T4 and H. influenze phage, I don't get any fine details like striations in the tail. Essentially I get nice looking hexagons sitting on top of tails but no fine structure. I have tried staining with 1% PTA or 2% uranyl acetate or vandium. I have tried 2 basic protocols:
1. putting a drop of the prep on a carbon coated grid for 30 to 120 sec, wicking the drop off, rinsing with a drop of water, and then adding a drop of stain and almost immediately removing it. Skipping the rinse doesn't seem to help nor does lengthing the staining time.
2. putting a 5 ul drop of the prep on the grid and then adding a 5 ul drop of stain.
Are there any negative staining experts out there with hints for getting the best image?
TIA, tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Pat Hales wrote: ================================================== I have been asked to find out if anyone can share any information about silicon nitride or silicon oxide supports for TEM. We are interested in 100 nm thick plates or sheets - where to get them or how to make them; and on my part, how they are used. ================================================= Most of the answers to the questions are found on our website URL http://www.2spi.com/catalog/instruments/silicon-nitride.html
The requested thickness is one of our standard thicknesses, and the short answer is that they are used just like a normal TEM grid since the outer support silicon is 3 mm diameter and can fit into any standard 3 mm grid holder. Depending on the type of work you are doing, you might want to consider something thinner than 100 um.
The properties of Si3N4 in this thickness are such that there is no such thing as a free standing "sheet" since at that thickness it would instantly curl up on itself into a roll.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
} Could something be sent to this list too please - it would certainly } interest me. } } thanks } } Malcolm Haswell
Malcolm,
This is the text of the article from the May '97 Microscopy Today. The U. =46lorida web site and Shirley Pinchuck mentioned in Robin Cross' post will have more information.
Phil
HMDS and Specimen Drying for SEM:
Hexamethyldisilizane (HMDS) is an excellent method of chemical drying of hydrated specimens. There are several variables involved in its use, the most easily controlled being the number of transitional steps from 100% ethanol (EtOH) to 100% HMDS and the drying temperature. Fixation and dehydration are the same for both HMDS and CPD. Once the specimen is in the final 100% ethanol, it must then be transferred to 100% HMDS through a graded series of ethanol-HMDS mixtures. This can follow one of four basic paths:
Ratio absEtOH : HMDS starting from 100% EtOH going to 100% HMDS
1)100% E =3D} 1:1=3D} 100% H 2)100% E =3D} 2:1=3D} 1: 2=3D} 100% H 3)100% E =3D} 2:1=3D} 1:1=3D} 1: 2=3D} 100% H 4)100% E =3D} 3 :1 =3D} 1:1=3D} 1: 3=3D} 100% H (Extra gradations may be added as needed, for instance between the 3 :1 - 1:1 and 1:1 - 1: 3 steps)
After the final transitional step, make 3 changes in HMDS (the last two steps can sometimes be skipped). Dry from the last 100% HMDS step, or exchange with new 100% HMDS one final time then dry. There should be just enough HMDS in the container to cover the specimen, any more just increases the drying time. Time in these steps will usually be the same as that used in the final 100% EtOH steps. However, the time can apparently be extended will little ill effect on the sample. Incomplete transition from EtOH to HMDS is a worse source of problems than extended time in HMDS. Choice of steps is basically determined by sample density, and the permeability (to HMDS and ethanol) of the least permeable structures in the sample. Microorganisms can usually be done with the first series, animal tissues need the second or fourth series, and plants require the fourth series, or even more gradations because of their cell walls.
Drying is done at either: 25=BA C =3D room temperature 8 hr =3D} overnight 37=BA C } 45=BA C }1=3D} 4hr 60=BA C } (Drying time by both temperature and volume of fluid.) The higher the temperature, the shorter the drying time, but the quality of results may also vary. Microscopic unicellular algae did best at 60=BA C, fish skin at room temperature, bacteria equally well at room temperature and 60=BA C. HMDS may have a significant time advantage over CPD. If more specimens are being processed that can fit in the CPD chamber, then the times in the transitional fluids and for drying will be much less than the time necessary for CPD. The greater the number of samples that can be batch processed, the greater the time advantage for HMDS. HMDS has another advantage: if you can find containers that seal tightly enough (HMDS likes to evaporate given any chance at all), samples can be collected in the field, processed to 100% HMDS, then stored in vials filled with HMDS and transported long distances from remote sites - like from Antarctica to Chicago, Illinois. The samples are protected by the fluid, and at least for some specimens, so fewer artifacts than specimens stored in fixatives or alcohol. Dried specimens are of course fragile. HMDS is not the cure-all for specimen drying. It can introduce it's own distorting artitfacts and some samples, biological ones in particular, still shrink after drying as they do with CPD or freeze-drying. Some specimens do poorly when dried from HMDS. Plant tissues in particular may not do well, and may be better off dried by CPD. Also, if the specimen is going to be studied for elements that are labile, or in solution, standard fixation and dehydration methods won=EDt work. Cryo techniques must be used, and if the specimen is to be examined in an unfrozen state, for example to look at structures and elements within cavities that would be obscured by ice if left hydrated, then the specimen must be dried by freeze-drying methods. All of this information is empirical. Theoretical explanations and any modifications for particular samples are welcome! A final note: HMDS must be used in a flume hood! A sniff of it will clear the sinuses back to the foramen magnum.
--MT
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net
Instead of etching the Si off to prepare a Plan View TEM sample, I suggest mechanically thinning the Si substrate. Allied High Tech is the manufacture of a tool, the MultiPrep, which is a semi-automatic tool for preparing SEM, TEM cross sections and TEM plan view samples in addition to its other capabilities.
I am an employee of Allied and have an interest providing you with this information. If you would like to receive more detailed information please contact me or visit our website http://www.alliedhightech.com
Sincerely,
Ed Hirsch
} Hi, } } I was wanting to know if anyone has used any wax or etch resist } which will protect against warm KOH. I need to mask a Si/SiO2/Si } wafer (Silicon on Insulator) to preferentially remove a the thick Si } backside of my specimen and leave the surface Si/SiO2 bilayer intact } for TEM imaging. I was going to use warm KOH (50 C) to remove the Si } but I need to mask the surface and define a window on the backside Si } to etch through (the KOH will stop at the SiO2 layer). Any } suggestions? } } John } --------------------------------------- } John Michael Glasko } Materials Science and Engineering Dept. } 2142 Burlington Labs/Yarborough St } Raleigh, NC 27695-7916 } USA } tel: (919)-515-7217, fax: (919)-513-1699 } jmglasko-at-unity.ncsu.edu } } ************************************************* Edward A. Hirsch Product Application Specialist Allied High Tech 2376 East Pacifica Place Rancho Dominguez, CA 90220 ph: (919) 846-9628 vm:(800)675-1118 x245 fx: (310)762-6808 http://www.alliedhightech.com *************************************************
I need information regarding use of low vacuum SEM in analysis of tissue culture cells combined with immunogold labelling. Has anyone tried? Most of the images I've seen from the manufacturer's brochures are of algae, insects or foods. I'd appreciate comments.
Corazon D. Bucana UT M.D. Anderson Cancer Center Houston, Texas
} 1. putting a drop of the prep on a carbon coated grid for 30 to 120 } sec, wicking the drop off, rinsing with a drop of water, and then adding a } drop of stain and almost immediately removing it. Skipping the rinse } doesn't seem to help nor does lengthing the staining time.
} 2. putting a 5 ul drop of the prep on the grid and then adding a 5 ul } drop of stain.
a) Did you 'glow discharge' the grid shortly before you adsorbed the drop? b) Have you tried Uranyl Formiate solution? (I think 0.75%w/w and pH around 4 should be good.) c) Have you tried lower concentration of Uranyl salt? Maybe the fine details are embedded in a thick salt layer so that you don't get any contrast there.
The procedure I normally use is: - prepare parafilm with at least 2 droplets of water (or sometimes buffer?) and another two of stain solution - glow discharge the grid for around 10 to 20 s (blue-white) - clamp it into tweezer - put 5um drop on the grid and wait... whatever.. 30 s, 60 s.... - touch with filter paper to soak solution away (blotting) - wash on first drop of water by touching the surface, blot again - repeat above step at least once - then go to stain droplet and do the same - the second stain droplet is then used to do the staining procedure, so touch the surface and wait for a few seconds (lets say 10 to 20) - blot a last time and that's it
You may not need all these steps, but I have been very successful with this technique and don't see any reason not to do it this way. If you don't have Uranyl Formiate, you can try the upper protocol with Uranyl Acetate (2% or lower). My experience is that usually it should work as nicely as UFo, but I was told that UAc does 'melt' more strongly and more quickly in the electron beam. I am looking forward to other recipies. I am very sure that you can find as many recipies as there are microscopists...
By the way, I have never been successful using PTA. Even with very sensitive proteins which don't like a low pH, I always had the best results in using Uranyl solutions.
Best regards,
Bettina
*** Bettina Wolpensinger Electron Microscope Unit University of New South Wales Sydney, NSW 2052, AUSTRALIA phone: +612 9385 6390 fax: +621 9385 6400 b.wolpensinger-at-unsw.edu.au http://srv.emunit.unsw.edu.au ***
Pale yellow colored Schiff-solutions are usable as long as they are not pink colored and control slides are stained as they should be...=20
Slightly orange color is probably produced by impuritys in the dye (acrid= in orange)... Some authors call for "basic fuchsine for schiff's" or "guaranteed acridin orange free pararosanilin" instead of basic fuchsin.
Basic fuchsin should be DISSOLVED COMPLETELY before adding the sodium metabisulfite and the HCl. A lot of recipes for Schiff's call for solutio= n of 1 gm of basic fuchsin in "HOT WATER" (80 - 100=B0C). Be sure to CLOSE = THE CONTAINER TIGHTLY after adding the sodium metabisulfite and the HCl and let stand for a few hours (some auteurs say "18 -24hrs") IN THE DARK. (Sorry: I'm not shouting, only emphasizing...).
The solution should have a strong "sulfur" smell.
I never had problems in preparing Schiff's. I use Basic fuchsin Merck "Certistain" and reagents UCB "PA" grade...
Several years a go our lab switched from the traditional KPTA to NHPTA. which enhances the fine structural details often lost with more aggressive stains. Prepare a 1% solution of phosphotungstic acid , then pH the stain with 1N ammonium hydroxide to 6.5 and / or 7.0. Regards, Skip
If you can't find a suitable resist for the KOH solution, I suggest dimpling the back (the Si) side most of the way through and then etching for a short time in KOH till the remaining thin region of Si is gone, but the surrounding thick support layer is mostly intact.
This should work very well in theory, though I've never done it myself.
For dimplers, if you have a choice, I recommend the D500i (or subsequent models) from the VCR Group, Inc.. It has exquisite thickness control, very good damping against vibration (invaluable with brittle samples), imbedded diamond abrasive wheels, and can be set up to produce a large flat bottomed dimple. (I've evaluated all makes before our purchase, and the VCR unit is by far the best - I have no interest or relationship with the manufacturer.)
If you have difficulty locating a dimpler, you can try ours, but we are short of man-power so you would have to travel and fiddle with it on your own. A better alternative might be to talk to the VCR applications lab and have them make a few samples for you as a way of demonstrating the instrument's capabilities. When I initially dealt with them, they were very accommodating. Unfortunately, I do not have the URL for the company, but an internet search should be fruitful with that.
An alternative technique might be a chemical jet polish, where the area of sample impacted by the jet of the etching solution thins preferentially. But that's a lot more tricky to control, and your remaining insoluble film must be able to withstand the pressure of the jet.
Whichever way you succeed, I would be interested what method(s) ended working for you.
Best of luck,
Valdemar rwafu-at-bsco.com or valdemar-at-fast.net
Valdemar Furdanowicz, Ph.D. Bethlehem Steel Co. Research G-165 Bethlehem, PA 18016-7699
Bethlehem -----Original Message----- } From: Edward Hirsch {edhirsch-at-att.net} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I have a Tracor Northern 8502 (5500/5700 hybrid) imaging system which is coupled with an ISI (now Topcon?) WB-6 SEM. I would like to transfer = the x-ray dot maps, x-ray spectra, and SEM images from the TN8502 to a=20 Win98 PC (TIFF, BMP, etc...). The 5700 side (imaging) runs on = Microware's=20 OS-9 and the 5500 (x-ray) side runs on a modified Fortran called = "flextran".
Please let me know if you have already solved this problem! Thanks in=20 advance for any suggestions.
Brian Reid Department of Chemistry University of Texas at Dallas 972-883-2709 breid-at-utdallas.edu
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I have a Tracor Northern 8502 = (5500/5700 hybrid)=20 imaging system which is {BR} coupled with an ISI (now Topcon?) WB-6=20 SEM. I would like to transfer the {BR} x-ray dot maps, x-ray = spectra,=20 and SEM images from the TN8502 to a {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Win98 PC (TIFF, BMP, etc...). = The 5700=20 side (imaging) runs on Microware's {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} OS-9 and the {/FONT} {FONT = color=3D#000000=20 size=3D2} 5500 (x-ray) side runs on a modified Fortran called=20 "flextran". {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Please let me know if you have = already solved=20 this problem! Thanks in {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} advance for any = suggestions. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Brian Reid {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Department of Chemistry {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} University of Texas at = Dallas {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} 972-883-2709 {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {A=20 href=3D"mailto:breid-at-utdallas.edu"} breid-at-utdallas.edu {/A} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 = size=3D2} {BR} {/FONT} {/DIV} {/BODY} {/HTML}
I'm no expert but we have had problems because of the following:
My experience with UA is that when it works it's great but it doesn't always work.
With PTA a bad batch of PTA stock could result in poor images - especially if it's old or not really a good quality reagent..
Do you adjust the pH of your PTA? It is normally fairly acid and should be adjusted to between 6 and 7 but you can experiment. We have always understood that it is best to use potassium hydroxide and not sodium hydroxide to adjust - personally I've seen little difference.
If you are using carbon coatings on your grids and clean preparations of virus maybe you need a little BSA to help spreading (not normally a problem with our samples).
If none of the above work it might be worth trying some of the newer more expensive stains like sodium silicotungstate or methylamine tungstate which can give nice results.
I suspect you may have tried most of the above, but I hope that something will help. Good Luck.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
Disclaimer - all my own experiences and thoughts and it's mainlty black magic anyway,
---------- } From: Tom Phillips To: Microscopy
I have gotten some pretty good TEM's of a negative stained bacteriophage prep but, unlike some published photos of T4 and H. influenze phage, I don't get any fine details like striations in the tail. Essentially I get nice looking hexagons sitting on top of tails but no fine structure. I have tried staining with 1% PTA or 2% uranyl acetate or vandium. I have tried 2 basic protocols:
1. putting a drop of the prep on a carbon coated grid for 30 to 120 sec, wicking the drop off, rinsing with a drop of water, and then adding a drop of stain and almost immediately removing it. Skipping the rinse doesn't seem to help nor does lengthing the staining time.
2. putting a 5 ul drop of the prep on the grid and then adding a 5 ul drop of stain.
Are there any negative staining experts out there with hints for getting the best image?
TIA, tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
by tartan.richmond.edu (8.8.8/8.8.8) with ESMTP id KAA16915 for {Microscopy-at-MSA.microscopy.com} ; Fri, 13 Nov 1998 10:15:09 -0500 (EST) X-Sender: gradice-at-facstaff.richmond.edu Message-Id: {l03130304b271fce4d6c6-at-[141.166.55.15]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Thanks all for your tips on preparing Schiff's reagent. Here are most of the suggestions, except for a couple I inadvertently deleted. My original problem was that my protocol said the reagent should be clear, while mine remained colored.
I tried making up the Schiff Reagent for the first time a few weeks ago and had some problems getting it to turn out as the book said it should (I was using Hayat 1993, but the recipe is the same). My problem was that Hayat said that after shaking overnight, the solution should be clear tan colored; it instead went from grape juice to wine colored. My Prof. said just to go ahead and add carbon (I used animal charcoal) and filter. The resulting solution was a clear pale pink. He said that was okay, just add a pinch more metabisulfite and cap tightly. I did and the staining worked well. He says the metabisulfite is the important thing.
} From: hank p adams {hpadams-at-bcm.tmc.edu}
Gary, that is not an uncommon problem. It depends on your source of basic fucshin which is if my memory serves me right is a combination of pararosaline dyes. Some sources are better than others and I went thru quite a few. I had absolutely perfect results with some produced from Sigma under pararosaniline in their catalogue, though I don't remember which salt I used. The solution came out perfectly clear and I stored aliquots at minus 20C and it seemed quite stable.
} From: bozzola-at-siu.edu (John J. Bozzola)
Gary, It has been awhile since I worked with the Schiff reagent but I used to mix up large batches of it. Firstly, the formula that I have (Lillie (1954) is slightly different:
basic fuchsin (C.C.) 1 gm (best to use a dye certified by the Biological Stain Commission) distilled water 80 ml NaHSO3 is 2 gm or 1.9 gm Na2S2O5 N HCl 20 ml
I noticed that this formula uses more bisulfite and more HCl -- both of which generate more SO2 for decolorization of the fuchsin.
This should go in a tightly stoppered container (to keep in the SO2 which does the decolorization of the fuchsin) and shaken every 10 minutes for a period of 2 hr. Add 500 mg finely powdered fresh charcoal (we used coconut charcoal). Charcoal must be finely ground (like talc). You can do this twice to remove any residual color. Sometimes I even added charcoal, shake and store in a refrigerator overnight. Filter through several layers of paper filter media. When colorless store at 5 deg C. If white precipitate shows up, it can be refiltered. Pink color means time to discard it.
Since your stain is not dissolving, something may be wrong with the fuchsin. Lack of destaining may be caused by not enough bisulfite or loose fitting container. Most often, we did not even need to use the charcoal.
} From: "C. John Runions" {cjr14-at-cornell.edu}
Brian asks ...
} ... I would like to transfer the } x-ray dot maps, x-ray spectra, and SEM images ...
I don't believe your system would use any brand of file compression, so the bitmap should be intact. If you know the bitmap's size ... e.g., 512by512by8bit, then a program which can open a "raw" bitmap will work .. e.g., "Photoshop-} File-} Open as" ... indicate "raw" and enter the bitmap size and have it guess at the header size. Because some filetypes might also include info at the end of the file, you may have to guess at the header size yourself 'til the bitmap rows and columns align properly.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
John, 1:4 HF:Nitric will do what you want. It etches SiO2, but slowly. As long as you have more than a hundred Angstroms or so of SiO2, you should be able to remove the substrate with no trouble. It will help if you mechanically thin from the back to 100 um or so; the solution should get through the silicon in under 5 mins. Sample agitation / rotation will help keep the surface smooth, although this isn't really important for your application since it will smooth anyway when it reaches the SiO2.
Cheers,
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK } } Hi, } } I was wanting to know if anyone has used any wax or etch resist } which will protect against warm KOH. I need to mask a Si/SiO2/Si } wafer (Silicon on Insulator) to preferentially remove a the thick Si } backside of my specimen and leave the surface Si/SiO2 bilayer intact } for TEM imaging. I was going to use warm KOH (50 C) to remove the Si } but I need to mask the surface and define a window on the backside Si } to etch through (the KOH will stop at the SiO2 layer). Any } suggestions? } } John } --------------------------------------- } John Michael Glasko } Materials Science and Engineering Dept. } 2142 Burlington Labs/Yarborough St } Raleigh, NC 27695-7916 } USA } tel: (919)-515-7217, fax: (919)-513-1699 } jmglasko-at-unity.ncsu.edu
There is a program from SAMx called TN2Win that we bought for this. It = will convert the spectra from TN format to ASCII x,y and EMSA format. I = forget the formats that the image files are stored in. There are some ramifications that you have to go through to do this. You have to have = a serial card on the TN5500 you have to boot the system to a remote = terminal which will be the PC. There are some cables that come witht he system = and both PC and TN software disks. The system was about $3500 if I = remember correctly. You can get info from their web site.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Brian Reid To: Microscopy-at-Sparc5.Microscopy.Com
Dear Michelle,
It is possible that your protein is present in very little amount in the plant tissue. You may try the same fixation protocol for immunocytochemistry but embed in paraffin. Make thick sections, label it and look at it under a fluorescent scope. You may use a confocal scope too and optically slice the tissue to see if you get any labelling at all. If you use a confocal scope then use Cy5 as a fluorochrome, it gives very little background fluorescence in leaf tissue.
Good luck,
Soumitra Ghoshroy Ph.D. Department of Plant Sciences University of Arizona 303 Forbes Building Tucson, AZ 85721 Tel: 520-621-1230 Fax: 520-621-7186 e-mail: ghoshroy-at-ag.arizona.edu
On Wed, 11 Nov 1998, Michelle L. Peiffer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Sorry that I didn't provide all the details on the original post, here they } are: } } We are attempting to label g-protein in Arabidopsis guard cells. Westerns } indicate the protein is water soluble, but also associates with membranes, } though it is not an integral membrane protein. Conventional fixation of } the cells, with aldehydes, osmium, embedded in Spurrs yeilds good } ultrastructure, but no labelling. For labelling we are fixing with 4% } paraformaldehyde, 0.5% glutaraldehyde in 0.1 M phosphate buffer; } dehydration in ethanol, embedding in LR White. The primary antibody is } polyclonal, affinity purified IgG, the secondary is goat anti-rabbit } conjugated to 10 nm colloidal gold. We are getting acceptable } ultrastructure and labelling in E. coli (expressing the protein) processed } this way, but not even a hint of labelling in plants. } } We do have the resources to do cryo-work, but the equipment is all brand } new, and we are still working out the details. Any suggestions on } improving this protocol will be greatly appreciated. Thanks } } } } #################################################### } Michelle Peiffer } Electron Microscope Facility for the Life Sciences } The Biotechnology Institute for Research and Education } 1 South Frear Lab } University Park, PA 16802 } 814-865-0212 email:mlk101-at-psu.edu } #################################################### } } }
Institution x Felix d'Herelle Reference Center for Bacterial Viruses, Department of x Microbiology, Faculty of Medicine, Laval University, Sainte-Foy, Canada. x
Title x A catalogue of T4-type bacteriophages. [Review] [115 refs] x
Source x Archives of Virology. 142(12):2329-45, 1997.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
In a message dated 98-11-11 10:13:55 EST, mlk101-at-psu.edu writes:
{ { We are attempting to label g-protein in Arabidopsis guard cells. Westerns indicate the protein is water soluble, but also associates with membranes, though it is not an integral membrane protein. Conventional fixation of the cells, with aldehydes, osmium, embedded in Spurrs yeilds good ultrastructure, but no labelling. For labelling we are fixing with 4% paraformaldehyde, 0.5% glutaraldehyde in 0.1 M phosphate buffer; dehydration in ethanol, embedding in LR White. The primary antibody is polyclonal, affinity purified IgG, the secondary is goat anti-rabbit conjugated to 10 nm colloidal gold. We are getting acceptable ultrastructure and labelling in E. coli (expressing the protein) processed this way, but not even a hint of labelling in plants.
We do have the resources to do cryo-work, but the equipment is all brand new, and we are still working out the details. Any suggestions on improving this protocol will be greatly appreciated. Thanks } } Hi Michelle,
If the protein is water soluble, cryoultramicrotomy is probably going to be the solution to your problems. This doesn't fully explain why you can label the protein in E. coli that are expressing it. Perhaps the bacteria are producing so much of the protein that by the time they are processed and embedded there is still enough left in a conformation in the cells that it can be resolved by immuno. If the protein is in much lower concentrations in the guard cells the processing may cause its complete extraction or denaturation.
Until you can get comfortable with the cryo setup, perhaps you could try either quick freezing and freeze-drying or chemical fixation followed by low temperature embedding in a polar resin like Lowicryl K4M. The low temperatures combined with a polar resin might prevent some of the extraction and denaturation. Lowicryl is available from Energy Beam Sciences (1-800-992-9037).
If you freeze-dry the specimens you can do a quick vapor fixation with osmium tetroxide in a sealed bell jar or chamber. Under these circumstances the osmium seems to bind to the specimens in a different manner than in aqueous chemical fixation, and it does not go through the "secondary osmium black" formation like it does when it is dehydrated with ethanol. But it will have a fixative effect and help with the ultrastructure. If the specimens are fixed with osmium vapor you should probably avoid Lowicryl as the resin, since the black color will interfere with Lowicryl's polymerization by UV light.
We have shown in several publications that you can quick freeze and freeze-dry (or "molecular distillation" dry) erythrocytes, follow this by vapor fixation with aldehydes and osmium, embed in Spurr resin ( ! ) and still get excellent labeling of carbonic anhydrase, which is notoriously water solubile *and* fixative-labile.
One note of caution: If you vapor fix with osmium in a partial vacuum be sure the vacuum pump has a cold trap on it. If you don't have a cold trap be sure to change the pump oil after the run. The osmium will get into the oil and turn it black. This is a very effective, but somewhat expensive, osmium trap!
If you aren't set up for freeze-drying or Lowicryl embedding or don't want to go to the trouble, perhaps there is someone on this listserver that does such things on a regular basis and would be willing to help. But cryoultramicrotomy is really the way to proceed with this, imho.
Good luck, let us know how things turn out.
Best regards,
Bob **************************************** Robert (Bob) Chiovetti Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging *****************************************
I was given an assignment to map sulphur using EELS, does anyone have any good tips to map sulphur? Like microscope conditions, sample thickness, energy spread, etc. I've been trying to do it several times, but couldn't see any sulphur edge. Although, I detected sulphur using EDX.
I'm using FEG-CM20 with the Gatan PEELS.
3.81KV extraction voltage 3mm or 5mm PEELS apperature Gun lens 5 Spot size 6. energy dispersion 0.5 or 0.3 ev/channel.
I can't see any sulphur edge, but after the power law background substraction in EmiSpec there seems to be a variation of the sulphur I was mapping, but my advisor doesn't feel comfortable with the data because the entire spectrum seems to be varying ... so I'm not sure if it was from the thickness variation or the actual sulphur variation in the mapping. Are we supposed to see that actual sulphur edge? ... The sulphur content within the sample I'm looking at is not that much.
Please advise me ... any tips would be greatly appreciated ... I seem to be loosing hope in seeing this sulphur edge. Sometimes I even get delusional in seeing these edge ...
This international conference will focus on the latest developments in the study of the structural and electrical properties of semiconductors by the application of transmission and scanning electron microscopy, scanning probe microscopy and X-ray techniques.
The state-of-the-art in all important subject areas will be addressed, including the characterisation of bulk and thin film as-grown materials, the study of lattice defect and impurity behaviour and the investigation of advanced semiconductor processing procedures.
Special conference sessions will concentrate on recent advances in high-resolution imaging and analytical eletron microscopy, the characteristics of epitaxial layers (including III-V nitrides), quantum wells, wires and dots, dislocation structure, the effects of device processing treatments (including, especially, advanced silicon technology) and metal-semiconductor contacts and silicides. Prominent invited speakers will introduce each topic area.
The Proceedings of the conference will be published and the final call for papers has now been issued.
The abstract deadline is 4 DECEMBER 1998, and full abstract submission information can be found at the conference Web site http://www.iop.org/Confs. Enquiries may also be directed to Jacqueline Watts at the Institute of Physics, UK, Tel: +44-171-470-4800, E-mail: conferences-at-iop.org.
Dear Colleagues: Could anyone of you kindly tell me the titles and the publishers of good books on confocal microscopy? I need the ones for beginners or as more extensive reference book. Thanks in advance.
Yuhui Xu, MD,PhD Chief, EM Core Facility DFCI, Harvard Medical School
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Tom,
In addition to the many recipes for stains you must be receiving, you = might also like to try a simple "low-dose" imaging protocol. I have found = when using negative stains that the image selection and focussing = procedures can damage fragile structures in the sample. Focussing on an = area close to a particle and then moving over to take the picture can help = prevent this. If you own a newer TEM, you might even have a low dose = capability built in. I have found this method very useful for small, = fragile samples, even when coated with thick films of aqueous uranyl = acetate or phosphotungstic acid.
Alternatively, the support film might be too thick to image the fine = detail. Is your grid carbon coated, as you post, or is it formvar/carbon = coated, which is more usual? Carbon alone offers more for high resolution = work.
Finally, have you checked out the vibrations affecting the microscope? = High resolution requires stable conditions.
Check out my web site for a brief summary of the options {http://www.hei.= org/htm/neg.htm} . =
I look forward to postings containing stain receipies. =
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
by imo14.mx.aol.com (IMOv16.10) id LISBa17681; Fri, 13 Nov 1998 18:14:36 -0500 (EST) Message-ID: {6c2b6ab4.364cbd5c-at-aol.com}
Hi Tom,
I would only add to Paul Webster's and other's comments that an amorphous carbon film that has been glow-discharged in a partial vacuum with air for about 60 seconds is pretty hard to beat when it comes to a substrate for negative staining. We used this procedure for T4 on one of my (numerous) postdocs in the Dept. of Microbiology at the Biozentrum in Basel. We never found anything to beat it. The stain spreads beautifully on the surface.
Bob **************************************** Robert (Bob) Chiovetti Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging *****************************************
We have a Tracor ADEM SEM which runs on the OS9 software from Microware. We use a program called OS9MAX to convert TIFF images from the ADEM (8500) to PC readable format. The only glitch is you have to have a PC that has a 5.25" floppy. You save the image in OS9 TIFF format onto a floppy on the 8500, then you have to put that floppy in a PC running OS9MAX. The OS9 image is converted to PC TIFF readable and can be used in Photoshop or whatever. We have only saved images so far, I'm not sure we can convert spectra with the program.
If you are interested, I'll look up the name of the company we purchased OS9MAX from and send you more info. The company is in Germany but had a turn- around time of a few days between the time we made the purchase and received the software. E-mail me for more details.
THANK YOU SO MUCH for all the responses and advises (and possibly future response and advice as well). I think I should put them in a summary and put the advises into practice as soon as possible.
Roseann asked about the type of specimen I'm working with:
The sample is a biological sample ... looking for the sulphur content in the particle which gives us skin color.
Larry and Yasuo mentioned about confirming the sulphur with EDS (it's the same with EDX, right?) And whether the peak that I thought was sulphur could be coming from Mo or Pb. And about the beam damage.
Frankly, I don't know that Mo and Pb peaks should be there and I would have to look into that, but I doubt it is because if I do see Pb peak it would be another peak together with the sulphur. My 2 advisors think, it's definitely sulphur. "Sulphur concentration is not that much" and I think what you said about EELS sensitivity is absolutely correct .. and that's my problem, I think my sulphur edge is hidden in the tail of the plasmon. I have been trying to do 2nd difference spectra myself, but my EL/P computer just died (I was too harsh on it; so now, I would have to fixed the computer first before I could start working on 2nd difference again.)
And about the 2nd difference ... do you think MacFac by Bonnet and Trebbia would be an equally good solution as well?
I'm still really new in this field and haven't been doing much reading (that's my really bad part)... so I don't really know how thick my specimen is in term of mean free path but I'll find out soon ... but the physical thickness was 70nm and I just got a new one that is supposed to be 30nm thick.
As for beam damage, I try to work with the new areas to see the sulphur edge with EELS first, then if I can't see the edge (which is always the case) I would confirm it with EDS (which EDS always show the sulphur peak).
The assignment was given to me partially as a challenge to test my ability to use EELS. And to do EDS spectrum mapping would take a very long long time, so that's why my advisor wants me to do EELS spectrum mapping.
BTW, has anyone converted the EmiSpec file into EL/P file? Does anyone know where I could get the header file to convert Emispec file into EL/P file? I have the header file for Emispec file, but I don't know exactly where I could get the header file for EL/P. All the software in my lab is the first version (most primitive). :P
All-in-all THANK YOU so much ... and I would to put all the advises into good use. Below is Gerd's advice if anyone else is interested.
Thank you, Ad
+++++++++
} From Gerd:
First I would collect two spectra with good counting statistics in the sulphur and in the low loss region. Splice them and do the single scattering deconvolution using the LOG-ratio method in EL/P. This ensures that you get always the same background before the edge and you cancel out thickness effects.
Secondly, if you have regions without sulfur and without, you can use the spatial difference technique: author = {H. M\"ullejans and J. Bruley}, title = {Improvements in Detection Sensitivity by Spatial--Difference Electron Energy--Loss Spectroscopy at Interfaces in Ceramics}, journal = {Ultramicroscopy}, year = 1994, volume = 53, number = 4, pages = {351-360}, You need two spectra which are taken under the same conditions and at locations with the same thickness. Then you use the one without for an improved background subtraction.
I want to do freeze fracture and replication of bacteria and viruses. I am using the Balzers Freeze Etching System BAF 400. I managed to get to the state of floating off the replicas on a liquid surface. But this is accompanied by some problems and by using water they don't float off properly.
Should I use another solution for floating off the replica?
What about the influence of the way the specimen carriers have been cleaned?
There are different kinds of specimen carriers some do have a central boring and some do not. I have tried different kinds of combinations, but couldn't yet work out a difference. Are there any hints on this?
I ended up with quite a bit of ice on the cooled parts in the chamber which is floated with air. Is it highly recommended to use dry nitrogen instead?
Is there a special way to bake out the system after doing a freeze-fracture?
A final question for BAF specialists. Whenever the system switches to 'high vac' the valve located at the bottom of the chamber 'leaks' what means that it allows air to blow out (I think it's the Plate Valve PVA 160P). This doesn't effect the operation of the system, it just results in a nearly continuous working mode (+noise) for the compressor providing the high pressure for pneumatic system. Is this normal, or is there anything I can do against?
I am looking forward getting your replies!
Regards,
Bettina
*** Bettina Wolpensinger Electron Microscope Unit University of New South Wales Sydney, NSW 2052 AUSTRALIA phone: +612 9385 6390 fax: +621 9385 6400 b.wolpensinger-at-unsw.edu.au http://srv.emunit.unsw.edu.au ***
I believe that one of the other listers has already mentioned molybdenum peaks. This has been a chronic problem on our Hitachi H7000 because of the Mo in the movable objective apertures.
We have had problems with EDX, not EELS, but I would expect that the geometry may be worse in EELS because your detector might more directly "see" objective apertures and so increase the chance of picking up Mo. If your system will pick up higher energies of Mo it would certainly be worth doing a quick check.
Incidentally I received a lot of advice about our molybdenum problem some time ago and I can't remember if I thanked everyone - so thanks, just in case. Unfortunately all we ever managed to do was minimize the effect, unless we removed the aperture rod which produced its own problems.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: Daraporn Arayasantiparb To: Microscopy
Dear Listers,
I was given an assignment to map sulphur using EELS, does anyone have any good tips to map sulphur? Like microscope conditions, sample thickness, energy spread, etc. I've been trying to do it several times, but couldn't see any sulphur edge. Although, I detected sulphur using EDX.
I'm using FEG-CM20 with the Gatan PEELS.
3.81KV extraction voltage 3mm or 5mm PEELS apperature Gun lens 5 Spot size 6. energy dispersion 0.5 or 0.3 ev/channel.
I can't see any sulphur edge, but after the power law background substraction in EmiSpec there seems to be a variation of the sulphur I was mapping, but my advisor doesn't feel comfortable with the data because the entire spectrum seems to be varying ... so I'm not sure if it was from the thickness variation or the actual sulphur variation in the mapping. Are we supposed to see that actual sulphur edge? ... The sulphur content within the sample I'm looking at is not that much.
Please advise me ... any tips would be greatly appreciated ... I seem to be loosing hope in seeing this sulphur edge. Sometimes I even get delusional in seeing these edge ...
by heinlein.acpub.duke.edu (8.8.5/Duke-4.6.0) with ESMTP id LAA03453; Mon, 16 Nov 1998 11:15:11 -0500 (EST) Received: (from saram-at-localhost) by godzilla2.acpub.duke.edu (8.8.5/Duke-4.6.0) id LAA19875; Mon, 16 Nov 1998 11:15:08 -0500 (EST)
James Pawley's Handbook of Biolobical Confocal Microscopy.
On Fri, 13 Nov 1998, Yuhui Xu wrote:
} Date: Fri, 13 Nov 1998 15:11:49 -0800 } From: Yuhui Xu {xuy-at-warren.med.harvard.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Books on Confocal microscopy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues: } Could anyone of you kindly tell me the titles and the publishers } of good books on confocal microscopy? I need the ones for beginners } or as more extensive reference book. } Thanks in advance. } } Yuhui Xu, MD,PhD } Chief, EM Core Facility } DFCI, Harvard Medical School } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} I want to do freeze fracture and replication of bacteria and viruses. I am } using } the Balzers Freeze Etching System BAF 400. I managed to get to the state of } floating off the replicas on a liquid surface. But this is accompanied by some } problems and by using water they don't float off properly. } } Should I use another solution for floating off the replica?
First you digest the sample, THEN you rinse with water. The "standard" is household bleach, but many samples will require something stronger. Have you read one of the basic books? } } What about the influence of the way the specimen carriers have been cleaned? } } Are there any hints on this?
Replicas usually are mounted on grids at the end of the cleaning process. Try moving the replica thru the sequence of cleaning solutions on a small glass ball. Make it by melting the tip of a disposable pipette. The replicas should float well. } } I ended up with quite a bit of ice on the cooled parts in the chamber which is } floated with air.
Not unusual; work faster.
Is it highly recommended to use dry nitrogen instead?
It helps!
} the valve located at the bottom of the chamber 'leaks' what means that it } allows air to blow out (I think it's the Plate Valve PVA 160P). This } doesn't effect the operation of the system,
Oh yes it does...
} Is this normal, or is there anything I can do against? } Get your system serviced! A Balzers specialist isn't necessary; anyone who understands vacuum systems will do. Ask your EM service person. }
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Readers. Re: the microscopits salary survey that we have been conducting, I regret to advise that only some 500 folks supplied their salary data. So, as we breakdown to education and experience (even in 3 year increments), there simply is not enough data in each category to provide any reasonable results. It seems that we would have needed twice the participation to come up with useful final data. Many thanks to all who contributed in our effort. Don Grimes, Microscopy Today
A while back there was a thread on the safe range of solvents to use when cleaning a polymer thin window. I volunteered to test more solvents if requested.
The only one requested was hexane. We succesfully used hexane to clean a window, and went further to soak it for 20 minutes (not recommended!) without causing degradation in the leak test. However after soaking there was a slight residue visible under the microscope. This was likely caused by contamination.
best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
I don't know what responses were made to Daraporn off the listserver, but there seems to be some confusion in what has been posted on the server.
Daraporn has seen a small peak at the S energy in his EDX spectra, but could not find S in the EELS spectrum. This could be for one of two reasons: 1) the S is not S at all, but Mo or Pb. In this case, with luck, he might expect to see the K-lines of Mo or the L-lines of Pb at higher energy, provided, of course, that there is enough present (the low-energy lines at the S energy are much more intense than the higher energy ones) Mo could certainly be coming from apertures, as Malcolm Haswell suggests. Pb could, conceivably, be coming from the x-ray shielding in the microscope, but do other CM20's have that problem? There is no obvious reason why Daraporn's should if others don't, unless they have made some modification to the stage area. It would seem unlikely that Mo or Pb were actually in the sample. EELS would be used to confirm the analysis because EELS is not susceptible to "hole count" effects. To use the Mo aperture as an example: the Mo x-ray signal is generated by stray radiation (scattered electrons, x-rays) hitting the large area of the mostly thick aperture material. Hence just a few electrons or x-rays can produce lots of Mo x-rays. In contrast, the few electrons that penetrate the aperture would have lost lots of energy and been scattered through large angles, so would not reach the EELS detector at all. There is no equivalent of the mechanism that generated the spurious x-ray signal.
Unfortunately, there is another possible reason for not seeing the S, even if it is really there. The sample could be too thick. The whole relationship of beam current, sample thickness, statistics and spatial resolution is very complex, both in EDX and EELS analysis. However, one point is probably worth mentioning: Comparisons of the detection sensitivities for EDX and EELS compare the results from the same samples. Typically, in practice samples suitable for EELS are thinner than those often used for EDX analysis. Hence, although theory might say that (at least up to Ca or so) the detection limits for EELS are better than those for EDX, this is only true for thin samples. In thicker samples, the EELS backround rises and the peak becomes blurred out, resulting in poorer detection, while in EDX you just get more signal and hence better detectability.
The papers (from NIST/NBS) that illustrate the sensitivity of EELS also point out the need for extremely high incident beam currents, available only in a LaB6 microscope. I don't know enough about the CM20 FEG to translate from the settings that Daraporn used to actual beam characteristics. However, it is clear that the optimum beam settings for EELS are very different from those for EDX, and also the spatial resolution is significantly degraded. It could be that his microscope conditions are not close to optimum for EELS.
Hi, This is a bit off the microscopy subject - my lab has a Dahle paper trimmer 551 and someone has damaged the blade. This has been a great little cutter for many years. I'm looking for information on a replacement (blades or the whole thing). Most of the microscopy companies sell the Rotatrim cutters...any opinions and product suggestions are appreciated. thanks, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
You're right, I can save the file into text file first to get the header file, thanks.
Dr. Thomas, you gave me a very tough question because I really don't understand many of the terms that was used (it's really my fault, cause I haven't been reading much)... please don't brush me off yet.
I don't know what is the approx. S-content ... they just told me "it's low" .. but with my parallel probe analysis with 100sec collection time, meter plate reading of 2.5 sec ... the maximum sulfur peak with EDS was ~1000 counts. But when I switch to do the analysis in STEM mode, because the probe is small, the sulfur peak has to be collected with a longer time. While with EELS, I only need 5 sec or something like that.
I need the mapping mainly for the cosmetic purpose and because it's give us spatially resolved information, something like that. That's the term they used.
You mentioned about "point spread in the detection system" in DigiPEELS, what is that? No one mentioned about that. Is that in the newer version?
Could you explain briefly how I could optimize EDS acquisition? What are the things to look for? I think I tried to have the shortest data acquisition time and high beam current already. Do you think 2.5 sec meter plate reading is still low? I'm really not too familiar with this.
_____________
Mo peak ... to Dr. Garber and Dr. Malcolm
I don't know about Mo aperture, but I'll check ... but I think the microscope I'm using has Be-window for EDS, is that the movable Mo-aperture you are talking about. Or is it the aperture of the microscope?
Thank you so much for all the responses, Ad Daraporn
We are interested in studying HE (high explosive) materials by SEM. Typical nonsenstive HE are HMX (octahydro-1,3,5,7 tetranitro-1,3,5,7 tetrazocine), RDX, TATB, PETN etc. Do you have any experience with or know of any literature, operating procedure, guidelines? Any contact? Please respond or call as appropriate? Thank you. Tri Tran Energetic Materials Section, Lawrence Livermore National Lab (925) 422-0915 tran4-at-llnl.gov
I agree with Sara. We bought a new Rotatrim a year ago and it works great.
Patty Jansma Tel:520-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
On Mon, 16 Nov 1998, Beth Richardson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } This is a bit off the microscopy subject - my lab has a Dahle paper trimmer } 551 and someone has damaged the blade. This has been a great little cutter } for many years. I'm looking for information on a replacement (blades or the } whole thing). Most of the microscopy companies sell the Rotatrim } cutters...any opinions and product suggestions are appreciated. } thanks, } Beth } } ************************************** } Beth Richardson } EM Lab Coordinator } Botany Department } University of Georgia } Athens, GA 30602 } } Phone - (706) 542-1790 } FAX - (706) 542-1805 } Email - beth-at-dogwood.botany.uga.edu } ************************************** } } }
If anyone needs a large green premiere paper cutter I have one. It is too big to use for what I do and would swap it for microscope goodies or books on microscopes or microtechnique the bummer is that you have to pick it up in Arizona it is a monster!
Does anyone have the website or e-mail address where I can find JEOL's y2K compliancy statement regarding the JEOL 5400 - scanning electron microscope. I know that the software is not compliant and have ordered the upgrade. I am however concerned about date related functions in the embedded system.
We are looking for spherical beads (metal, glass, etc) with a well defined diameter (range : 1 - 500 micron) and a small standard deviation. These beads will be used for testing particle analysis software in SEM.
The equipment we use is a Philips XL 30 FEG SEM and Noran Voyager software.
Any suggestion where to get these beads will be welcomed.
Thank you very much in advance.
Please respond directly to :
vanderwulp-at-pml.tno.nl
Kees van der Wulp Prins Maurits Laboratory - TNO POBox 45 2280 AA, RIJSWIJK (ZH) Netherlands
-----Original Message----- } From: Tri Tran [mailto:tran4-at-popsicle.llnl.gov] Sent: Monday, November 16, 1998 6:38 PM To: microscopy-at-sparc5.microscopy.com
We are interested in studying HE (high explosive) materials by SEM. Typical nonsenstive HE are HMX (octahydro-1,3,5,7 tetranitro-1,3,5,7 tetrazocine), RDX, TATB, PETN etc. Do you have any experience with or know of any literature, operating procedure, guidelines? Any contact? Please respond or call as appropriate? Thank you. Tri Tran Energetic Materials Section, Lawrence Livermore National Lab (925) 422-0915 tran4-at-llnl.gov
For the interest of microscopists in KwaZulu-Natal and beyond:
One of our overseas guests to the MSSA conference in December, Dr Alice Warley, will be visiting the Centre for Electron Microscopy at the University of Natal in Pietermaritzburg during late November. During her visit with us she will be conducting an informal workshop on the preparation of standards for EDX in the TEM. She will also present a talk entitled "Elelements inside Cells - Qualitative and Quantitative X-ray Microanalysis in Biology'.
We may be able to accommodate a very limited number of extra persons for the workshop. All interested persons are welcome to attend the talk at 15h00 on Thursday, 26 November.
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Private Bag X01, Scottsville 3209, KwaZulu-Natal, South Africa. Tel +27 (0)331 260 5155, Home +27 (0)331 962676 Fax +27 (0)331 260 5776 email: bruton-at-emu.unp.ac.za
Along these lines, we've just received a copy of a short booklet which outlines a number of tests addressing the Y2K problem. Further infomration is available from Info Check, LLC in Manassas, VA. Price: $19.95. We have an aol address for them at CMontana4-at-aol.com. (MME has no commercial interest in this endeavor).
Your best bet for this is an art supply store, or the UG art departments. If UG has a printing office, try them. Most of these cutters are built for printers and artists.
Phil
} Hi, } This is a bit off the microscopy subject - my lab has a Dahle paper trimmer } 551 and someone has damaged the blade. This has been a great little cutter } for many years. I'm looking for information on a replacement (blades or the } whole thing). Most of the microscopy companies sell the Rotatrim } cutters...any opinions and product suggestions are appreciated. } thanks, } Beth } } ************************************** } Beth Richardson } EM Lab Coordinator } Botany Department } University of Georgia } Athens, GA 30602 } } Phone - (706) 542-1790 } FAX - (706) 542-1805 } Email - beth-at-dogwood.botany.uga.edu } **************************************
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 oshel-at-terracom.net
I would be grateful if you would bring the available faculty position below to the attention of potential candidates.
thanks,
Nigel Browning
Faculty Position University of Illinois at Chicago
The Department of Physics at the University of Illinois at Chicago (UIC) is planning to fill one tenure-track assistant professorship position to begin Fall 1999. The department has a strong commitment to both undergraduate and graduate education and is dedicated to excellence in research. At present, we have thirty faculty members active in many areas of physics and biophysics. Our search will give special consideration to candidates in the areas of condensed matter physics and biophysics.
Successful candidates will have a Ph.D. degree or equivalent, are expected to develop top quality research programs and should show high promise of obtaining external research funding. We encourage research programs which take advantage of the numerous facilities in the Chicago area. Applicants should submit a vita and a statement of research interests and future plans. They should also arrange to have three letters of recommendation sent to: Prof. Inder P. Batra, Head, Department of Physics, University of Illinois at Chicago (MC 273), 845 W. Taylor St., Rm. 2236, Chicago, IL 60607-7059. Applications will be considered as they are received, with a deadline of March 25, 1999. The University of Illinois at Chicago is an Affirmative Action/Equal Opportunity Employer.
The National Institute of Standards and Technology sells polystyrene be= ads designed for particle sizing (SRM 1692). Information is available at http://ts.nist.gov/srm.
Joe Neilly Abbott Laboratories Dept. of Microscopy and Microanalysis Abbott Park, IL 60064-3537 =
Kees, Try the following. They may have what you need. We've purchased various microspheres from them with successful outcomes.
Bangs Labs, Inc. 9025 Technology Drive Fishers, Indiana 46038-2886 USA E-mail: info-at-bangslabs.com http://www.bangslabs.com
Good luck. Winston ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supervisor 11/17/98 11:53:38 AM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Does anyone have a procedure for distinguishing between hard and soft c= lay? Any information would be useful. Please don't hesitate to contact me o= ff line if you think this information would be of limited interest to the group=
Hair today, gone tomorrow I was recently asked by someone who develops hair color products, ( I didn't think I was greying that much) if I might be able to recommend a "system" that could be used to evaluate the efficacy of various hair colorants. Because this person would like to complete the examination without removing any hair, i.e. in-situ and in real time the "system" would have to be portable and consist of relatively high magnification optics coupled to a video camera with the signal fed to a color monitor. The desired magnification of such a system is 400 - 500 times. I am not familiar enough with the video camera market to identify/recommend a camera for such an application but am hopeful that someone out there can help.
Thanks, Paul Gerroir Xerox Research Centre of Canada
Is there a used SEM that requires relocation? We have few capital funds so we are looking for a second hand instrument that has out lasted its usefulness and needs a new home. If you have such a microscope within a reasonable distance to me and wish to negotiate terms and conditions of its removal, please contact me by e-mail below.
Best regards,
Gary
................................................................ Dr. Gary Faulkner, Ph.D. Director, Faculty of Medicine EM Facility Department of Microbiology & Immunology Sir Charles Tupper Building Dalhousie University Halifax, N.S. B3H 4H7 Tel: 902.494.2346 Fax: 902.494.5125 E-mail: Gary.Faulkner-at-Dal.Ca
How to improve the contrast in plant tissue included in Spurr resin? The double contrast with Uranyl acetate in ethanol solution and Lead citrate is not good for me. There is some special procedure for to solve this problem? With the ultra-low viscosity resin, the problem is worst (lower contrast than the Spurr resin). Thank's in advance.
M.Sc. Rinaldo Pires dos Santos Dept. of Botany - UFRGS Porto Alegre - RS - Brazil
with ESMTP id {01J4A9WJ7X9I8Y5EXU-at-denver.du.edu} for Microscopy-at-MSA.Microscopy.Com; Tue, 17 Nov 1998 15:30:50 MST Received: from localhost by du.edu (PMDF V5.1-10 #28062) with SMTP id {0F2L00N017W3YO-at-du.edu} for Microscopy-at-MSA.Microscopy.Com; Tue, 17 Nov 1998 15:31:15 -0700 (MST)
Hi,
We are having a problem retaining synaptic vesicles in tissue fixed with picric acid and 0.25% glut, embedded in LR Gold, and polymerized at -20 with UV. We will loose the antigen if we raise the percentage of glut. We cannot use osmium because we loose the antigen and then also cannot use UV for polymerization. Is there anyone who has been able to preserve vesicles with the above approach? Does anyone have any novel ideas? We will try tannic acid. Has anyone used this for vesicle preservation? I would so much appreciate any comments on what we are trying to do. Bye, Hildy {hcrowley-at-du.edu}
I really don't know how they made these slides back then. Check them out. It may be worth your while to look at these rare items..any info on how they were made would be appreciated. Thanks Dan Fleming
To view: http://cgi.ebay.com/aw-cgi/eBayISAPI.dll?ViewItem&item=42528374
This is not a forsale post but a request for information.
Frank - Please try the Home Page of the Clay Minerals Society: {http://shadow.agry.purdue.edu/clay/claymin/claymins.html} where you will find directions for subscribing to their list server. I'm sure someone there will be able to help you.
Being a clay mineralogist myself, I'll give it a try. Hard & Soft are commercial (common sense) terms meaning just that. Hard clays (such as "hard kaolin", also see "flint clay") require grinding and blending, wheras "soft kaolin" may require less preparation. Plastic clays are soft, wet and like modeling clay. Then there's "ball clay" which can be rolled up into a ball! Hard and Soft have no clear mineralogic or chemical meaning. See the AGI Glossary of Geology (1997?) for more info. Dave Pevear, Houston
FRANK KARL wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anyone have a procedure for distinguishing between hard and soft clay? } Any information would be useful. Please don't hesitate to contact me off line } if you think this information would be of limited interest to the group
for Microscopy-at-Sparc5.Microscopy.Com id AA24217; Wed, 18 Nov 98 16:00:20 +0100 Message-Id: {9811181500.AA24217-at-iris1.el.ub.es}
Dear colleagues,
We are working on the TEM characterisation of CuInS2 (usually called CIS) on glass substrates. We are preparing TEM samples out of these materials (both plan view and cross-section) and we are encountering problems with the preparation procedure, which are strong amorphisation of the CIS layer in plan-view and strong damage (not always amorphisation) in cross-section.
The preparation procedure we use is the standard preparation method we use for Si-based materials: For cross-section, we cut stripes out of the samples, glue them together. Next flat grinding, dimpling and finally ion milling are used. For plan-views, a piece of sample is ultrasonic cut and the preparation continues with the flat grinding, ... and ion milling only from the backside.
We believe that the problems we have are strongly related to the glass substrate and to the charging of the glass during ion milling, resulting in an overheating of the sample. This leads to extremely poor cross-sections and to even worst plan-view samples (however we still have some good results from few of these samples). It might be also that the CIS layer is strongly beam sensitive (any experience with it?).
Any help in trying to circumvent these problems will be extremely helpful and experience in preparing samples of the type glass substrate-thin layer would also be strongly appreciated.
By the way our ion milling machines can work at liquid nitrogen temperature in order to minimise sample heating, if this helps.
Thank you in advance for your answers.
Albert Romano-Rodriguez Dept. of Electronics Faculty of Physics University of Barcelona c/ Marti i Franques, 1 E-08028 BARCELONA Spain tel: +34-93-402 11 47 FAX: +34-93-402 11 48 e-mail: romano-at-el.ub.es
If not, I suspect that what is for sale is not a series of slides but a series of plates (hand painted pages to insert into a book) made by painting what was observed on a slide.
Gerard Turner, at Oxford, manages the collection of the Royal Microscopical Society. Either he or Dr. Savile Bradbury (Oxford, retired), would probably be the best sources for information on this sort of thing. Here in this country, Cecil Fox, who had a lot of contact with the Billings Collection at the Armed Forces Institute of Pathology, might know. Also, Don O'Leary manages the collection for the New York Microscopical Society. Try him at 201-797-8849.
You can probably contact either Gerard or Savile by writing/emailing Oxford University in the UK. I only have old contact information for Cecil Fox but would be happy to provide that information off-line if you are interested in talking to him.
Friends, I am interested in contacting some one who does micro CT imaging. If you are one or know of one I would appreciate hearing from you.
Greg Erdos Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
Responding to the message of {3651F0D1.6C4F-at-botanica.ufrgs.br} from Rinaldo Pires dos Santos {rinaldop-at-botanica.ufrgs.br} : } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear colleagues } } How to improve the contrast in plant tissue included in Spurr resin? The } double contrast with Uranyl acetate in ethanol solution and Lead citrate } is not good for me. There is some special procedure for to solve this } problem? With the ultra-low viscosity resin, the problem is worst (lower } contrast than the Spurr resin). } Thank's in advance. } } M.Sc. Rinaldo Pires dos Santos } Dept. of Botany - UFRGS } Porto Alegre - RS - Brazil
What kind of plant tissues?
1. Try a 50:50 mixture of Spurrs/Embed812 resins. Mix up resins seperately without their accelerators (DMAE & DMP-30). You can store them in the freezer. Then mix 50:50 just before use, mix in required amounts of the two accelerators, infiltrate and cure. You should get the better sectioning and staining properties of Embed812 plus the low viscosity of Spurrs.
2. Switch to Embed 812 100% and extend infiltration times.
3. What lead stain are you using? In my experience, Reynold's lead citrate doesn't seem to work too well with Spurrs. I use Sato's triple lead stain, preceeded by 3% UA, and get very good staining on plant and bacterial samples embedded in Embed812.
Good luck,
Gib
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Our group currently has a JEOL FX 2000 TEM available for sale to anyone interested. The system includes a cold and hot stage, PC driven X-ray system,and STEM attachment. Please respond if interested.
M.W. Rigler, Ph.D. MAS, Inc. Suwanee GA 770-866-3218
Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: from relay29.mx.aol.com (relay29.mail.aol.com [172.31.109.29]) by air09.mail.aol.com (v51.16) with SMTP; Thu, 12 Nov 1998 08:47:16 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by relay29.mx.aol.com (8.8.8/8.8.5/AOL-4.0.0) with SMTP id IAA00928; Thu, 12 Nov 1998 08:47:05 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA07680 for dist-Microscopy; Thu, 12 Nov 1998 07:22:03 -0600 Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id HAA07677 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 12 Nov 1998 07:21:32 -0600 } From: "Mriglermas-at-aol.com"-at-sparc5.microscopy.com Received: from imo17.mx.aol.com ([198.81.17.7]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA07670 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 12 Nov 1998 07:21:13 -0600 Received: from Mriglermas-at-aol.com by imo17.mx.aol.com (IMOv16.10) id NYLNa04133 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 12 Nov 1998 08:31:14 -0500 (EST) Message-ID: {d40385e2.364ae322-at-aol.com}
Our group currently has a JEOL FX 2000 TEM available for sale to anyone interested. The system includes a cold and hot stage, PC driven X-ray system,and STEM attachment. Please respond if interested.
M.W. Rigler, Ph.D. MAS, Inc. Suwanee GA 770-866-3218
To: Dan Fleming Re: Antique Microslides Dan, The expert on this subject is Dr. James B. McCormick. He has been assisted by M. Lamar Jones at many workshops during NSH conventions. You can get more info from National Society for Histotechnology at 301-262-6221.
Bob Santoianni Emory University Hospital Atlanta, Georgia robert_santoianni-at-emory.org
Kees - I've used 9.870 um (+/- 0.057 um) polystyrene spheres from Duke Scientific Corp with good success. I believe the company offers a large range in sizes as well. Their web site is www.dukescientific.com. Hope this is helpful.
Dave Joswiak University of Washington Dept. of Astrononmy Seattle, WA 98195 joswiak-at-astro.washington.edu
On Tue, 17 Nov 1998, Wulp, Cees van der wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear SEMmers, } } We are looking for spherical beads (metal, glass, etc) with a well defined } diameter (range : 1 - 500 micron) and a small standard deviation. } These beads will be used for testing particle analysis software in SEM. } } The equipment we use is a Philips XL 30 FEG SEM and Noran Voyager software. } } Any suggestion where to get these beads will be welcomed. } } Thank you very much in advance. } } Please respond directly to : } } vanderwulp-at-pml.tno.nl } } Kees van der Wulp } Prins Maurits Laboratory - TNO } POBox 45 } 2280 AA, RIJSWIJK (ZH) } Netherlands } }
I would suggest using a tripod polisher for getting the specimen to as thin as possible. This basically involves polishing the specimen to a thin wedge (between 0.5 - 3 degrees) using a set of polishing mats with varying grain size (30 um to 500 nm). I have polished silicon (mechanically) with this method to about 40 nm! But for most cases it will get you down to below 1um. The method uses a liquid to aid polishing lubrication, but you can use either oil based or water. I have a colleague who has used this to get his Barium based glass spcimens down to a couple of microns before finishing the polishing with a quick ion mill ( { 1 hr) and has report very good results. I have also used this specimen to map the Fe content in the crystallites in the glassy matrix.
The tripod polisher is produced by South Bay Technologies (sorry I do not have an address or phone) but you could try a web search.
I hope this helps. Yours, Jonathan Barnard Microstructural Physics, H.H.Wills Physics Lab. University of Bristol, U.K
Dear Paul, The last time I was at the MSC meeting, Chris Cathcart of Marivac had exactly the thing you are looking for, I believe. It was a little, portable video camera and lens system with built-in illumination. He held it up to his tie and we could see a brilliant image of the threads in the weave, in full colour. Contact him at Marivac, in Canada 1-800-565-5821. Good luck. You wrote:
} Hair today, gone tomorrow } I was recently asked by someone who develops hair color products, ( I didn't } think I was greying that much) if I might be able to recommend a "system" } that could be used to evaluate the efficacy of various hair colorants. } Because this person would like to complete the examination without removing } any hair, i.e. in-situ and in real time the "system" would have to be } portable and consist of relatively high magnification optics coupled to a } video camera with the signal fed to a color monitor. The desired } magnification of such a system is 400 - 500 times. I am not familiar enough } with the video camera market to identify/recommend a camera for such an } application but am hopeful that someone out there can help. } } Thanks, } Paul Gerroir } Xerox Research Centre of Canada } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
The person that would probably be the most helpful to you is Scott Walck at PPG, who is pretty well the King of cleaved glass TEM samples (Walck-at-ppg.com). He has developed a variation of the small-angle cleavage technique for preparing outstanding cross-sectional samples of glass or thin films on glass. This is a non-charging, heat-free preparation technique. A good reference is:
Scott D. Walck and John P. McCaffrey, "The small-angle cleavage technique: an update", Mat. Res. Soc. Symp. Proc. Vol. 480, Materials Research Society (1997) pp149-171.
I have an extra copy and will mail it, along with an instructional video of the small-angle cleavage technique that Scott and I made .
Cheers John
John P. McCaffrey Institute for Microstructural Sciences National Research Council of Canada M-50 Montreal Rd. Ottawa, Ontario K1A 0R6 Canada
---------- } From: Albert Romano-Rodriguez To: Microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------.
Dear colleagues,
We are working on the TEM characterisation of CuInS2 (usually called CIS) on glass substrates. We are preparing TEM samples out of these materials (both plan view and cross-section) and we are encountering problems with the preparation procedure, which are strong amorphisation of the CIS layer in plan-view and strong damage (not always amorphisation) in cross-section.
The preparation procedure we use is the standard preparation method we use for Si-based materials: For cross-section, we cut stripes out of the samples, glue them together. Next flat grinding, dimpling and finally ion milling are used. For plan-views, a piece of sample is ultrasonic cut and the preparation continues with the flat grinding, ... and ion milling only from the backside.
We believe that the problems we have are strongly related to the glass substrate and to the charging of the glass during ion milling, resulting in an overheating of the sample. This leads to extremely poor cross-sections and to even worst plan-view samples (however we still have some good results from few of these samples). It might be also that the CIS layer is strongly beam sensitive (any experience with it?).
Any help in trying to circumvent these problems will be extremely helpful and experience in preparing samples of the type glass substrate-thin layer would also be strongly appreciated.
By the way our ion milling machines can work at liquid nitrogen temperature in order to minimise sample heating, if this helps.
Thank you in advance for your answers.
Albert Romano-Rodriguez Dept. of Electronics Faculty of Physics University of Barcelona c/ Marti i Franques, 1 E-08028 BARCELONA Spain tel: +34-93-402 11 47 FAX: +34-93-402 11 48 e-mail: romano-at-el.ub.es
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE: Synaptic Vesicle Preservation??????? Dear Hildy,
Are you sure your problem is with retaining the synaptic vesicles? Could = it be more a problem of contrast? As I know it is possible to visualize = synaptic vesicles in brain fixed with 4% formaldehyde alone, I will assume = your problem is contrast. Here are a few suggestions.
1. Check to see if your antigen is present and the antibody works. Try = the labeling by light microscopy first (cryostat sections will work well = for this). If you can't get a signal by light microscopy go back to = understanding more about your antigen and the antibody. =
2 Check LR White labeling by light microscopy. If you can get a signal = in cryostat sections then use the same fixation protocol to embed the = tissue in LR WHite. A good protocol is to fix the tissues in 4% = formaldehyde alone. If the LM labeling only works after methanol fixation,= try that for the EM too. Mount the resin-embedded sections on glass and = label them for light microscopy. Ideally, you should be able to compare = both the antibody labeling protocol you use for routine LM labeling with = as much of the EM labeling protocol you can use. Silver enhancement = protocols for colloidal gold are useful here. If you can't see a signal, = it may be that the LR White is affecting the antigen-antibody interaction. = There is no way you will be able to perform EM until the LM conditions = have been worked out.
3. Improving contrast in LR White. If you can get the antibodies to label = LR White sections lable the sections for EM. You should see a signal. If = you don't see a signal go back to step 2. If you do see a signal, the = antigen is there. If you don't see the structure that should label, then = your problem is with section contrast. There are many ways to manipulate = this. = a) cut thicker sections. That sometimes helps. = b) examine the section more. This will burn out resin from the section = and may help. c) extract the tissue prior to embedding. Although detergents, low = osmolarity fixatives and modified buffers can have the effect you need (= less cytoplasm) be careful that your antigen is not extracted too. = Knowing as much about the antigen as possible is useful here. d) treat the tissue with osmium tetroxide or uranyl acetate prior to = embedding. Who says osmium tetroxide will cause you to lose your antigen = or stop you using UV polymerization? A good way to incorporate = contrasting agents is to freeze substitute a frozen sample on dry ice in = medium containing up to 1% OsO4 or uranyl acetate. At the lowered temperat= ure there is no production of the black reaction product typical of osmium = treatment. Make sure that OsO4 is washed away while the tissue is still = on dry ice. Want a protocol? try this: {http://www.hei.org/htm/pmfs.htm} . = Be brave.
4. Use cryosections for immunolabeling. If after all your attempts to get = a positive signal you get nothing, it may be that the resin is blocking = the antibody-antigen interaction. This sometimes happens. It may be time = for you to check out the modern world of cryosectioning. Technology and = specimen preparation protocols have improved enormously over the last few = years and make cryosectioning an almost routine technique. Check it out: {= http://www.hei.org/htm/cryo.htm}
Please feel free to contact me off-line if you need clarification of these = points (I did leave out many details). I am still sitting in an empty lab = (no TEM, no ultramicrotome etc!) so have more time to spare than usual.
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/apw.htm
HILDEGARD CROWLEY wrote:
} Hi, } } We are having a problem retaining synaptic vesicles in tissue fixed with } picric acid and 0.25% glut, embedded in LR Gold, and polymerized at -20 } with UV. We will loose the antigen if we raise the percentage of glut. = We } cannot use osmium because we loose the antigen and then also cannot use = UV } for } polymerization. } Is there anyone who has been able to preserve vesicles with the above } approach? } Does anyone have any novel ideas? We will try tannic acid. Has anyone } used this for vesicle preservation? } I would so much appreciate any comments on what we are trying to do. } Bye, } Hildy } {hcrowley-at-du.edu} } } } } } RFC822 header } ----------------------------------- } } Received: from Sparc5.Microscopy.Com [206.69.208.10] by mailhouse.hei.= org } (SMTPD32-4.07) id AEDC1EBF02A2; Tue, 17 Nov 1998 21:45:00 PST } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.= 11) = } id QAA11965 for dist-Microscopy; Tue, 17 Nov 1998 16:39:58 -0600 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id QAA11960 for = } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 17 Nov 1998 16:39:26 -= 0600 } Received: from atlas.cair.du.edu (atlas.cair.du.edu [130.253.2.202]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id QAA11947 for = } {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Nov 1998 16:39:08 -0600 } Received: from odin.cair.du.edu by denver.du.edu (PMDF V5.1-12 #28064) } with ESMTP id {01J4A9WJ7X9I8Y5EXU-at-denver.du.edu} for } Microscopy-at-MSA.Microscopy.Com; Tue, 17 Nov 1998 15:30:50 MST } Received: from localhost by du.edu (PMDF V5.1-10 #28062) } with SMTP id {0F2L00N017W3YO-at-du.edu} for Microscopy-at-MSA.Microscopy.Com; = Tue, } 17 Nov 1998 15:31:15 -0700 (MST) } Date: Tue, 17 Nov 1998 15:31:15 -0700 (MST) } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } Subject: Synaptic Vesicle Preservation??????? } X-Sender: hcrowley-at-odin.cair.du.edu } To: postmessage {Microscopy-at-Sparc5.Microscopy.Com} } Message-id: {Pine.OSF.3.95.981117151619.28840A-100000-at-odin.cair.du.edu} } MIME-version: 1.0 } Content-type: TEXT/PLAIN; charset=3DUS-ASCII } Content-transfer-encoding: 7BIT } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-UIDL: 907881491 } Status: U } =
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/apw.htm
by post2.fast.net (8.8.8/8.8.5) with SMTP id NAA04968; Wed, 18 Nov 1998 13:45:05 -0500 (EST) Message-ID: {000d01be1323$3091d7c0$02f993cd-at-rfurdanowiczw.hrl.bsco.com}
Materials Characterization Facility University of Minnesota
The Center for Interfacial Engineering at the University of Minnesota is = seeking an instrument specialist as a staff member in its materials characterizat= ion =
facility. The facility houses 5 EM's, 6 XRD/SAXS instruments, several SPM= 's, =
micro-indentors, and other surface and thin-film analytical instruments. = See our website for details (http://resolution.umn.edu). The person will work mai= nly in =
the EM laboratories. The principle responsibilities of the position incl= ude =
training researchers to operate transmission and scanning electron micros= copes, =
maintaining and operating the TEM=D5s (Philips CM30 and JEOL1210), and as= sisting =
users in TEM specimen preparation and data interpretation. The position r= equires a Ph.D. in biosciences, materials science, physics or related discipline.= Very =
strong hands-on experience in various TEM techniques and their applicatio= n to =
materials characterization is required. Applicants should also have the =
experience and flexibility to work with other techniques. Experience in s= pecimen preparation and working in a multi-user facility is particularly desirabl= e. This is an annually renewable professional appointment; 12 month, 100% time re= gular =
appointment with excellent university benefits. Position and salary will= be =
commensurate with education and experience.
Please send resume, three letters of recommendation and salary requiremen= ts to =
Elizabeth Guldan, Search Committee, Center for Interfacial Engineering, =
University of Minnesota, 187 Shepherd Labs, 100 Union St. SE, Minneapolis= , MN =
55455. Screening will begin on January 31, 1999 and end when a suitable =
applicant is identified.
The University of Minnesota is an Equal Opportunity Educator and Employer= =2E
__________________ Stuart McKernan stuartm-at-tc.um= n.edu Microscopy Specialist Office:(612) 626= -7594 CIE Characterization Facility, University of Minnesota Desk: (612) 624= -6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624= -6590
Albert: I am not familiar with the mechanical characteristics of CIS, but I wonder if you would do better trying tripod polishing and maybe avoid having to do ion milling ??.
Jordi Marti
Dear colleagues,
We are working on the TEM characterisation of CuInS2 (usually called CIS) on glass substrates. We are preparing TEM samples out of these materials (both plan view and cross-section) and we are encountering problems with the preparation procedure, which are strong amorphisation of the CIS layer in plan-view and strong damage (not always amorphisation) in cross-section.
The preparation procedure we use is the standard preparation method we use for Si-based materials: For cross-section, we cut stripes out of the samples, glue them together. Next flat grinding, dimpling and finally ion milling are used. For plan-views, a piece of sample is ultrasonic cut and the preparation continues with the flat grinding, ... and ion milling only from the backside.
We believe that the problems we have are strongly related to the glass substrate and to the charging of the glass during ion milling, resulting in an overheating of the sample. This leads to extremely poor cross-sections and to even worst plan-view samples (however we still have some good results from few of these samples). It might be also that the CIS layer is strongly beam sensitive (any experience with it?).
Any help in trying to circumvent these problems will be extremely helpful and experience in preparing samples of the type glass substrate-thin layer would also be strongly appreciated.
By the way our ion milling machines can work at liquid nitrogen temperature in order to minimise sample heating, if this helps.
Thank you in advance for your answers.
Albert Romano-Rodriguez Dept. of Electronics Faculty of Physics University of Barcelona c/ Marti i Franques, 1 E-08028 BARCELONA Spain tel: +34-93-402 11 47 FAX: +34-93-402 11 48 e-mail: romano-at-el.ub.es
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Hi,
I wonder if I could solicit advice off-line about the benefits and = drawbacks of currently available critical point dryers and high resolution = sputter coaters? All replies will be treated as opinions and will remain = confidential. =
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/apw.htm
Thanks to all responses. I think I have quite a number of things I would need to do and adjust and look into in my experiment before I could get back to all of you. All responses and tips and advises were very helpful. I'll put all of them into practice and pray for a better results this time.
Does anyone care for me to put all responses into a summary?
Special thanks to
Dr. Larry Thomas Dr. Malcolm Haswell Dr. Tony Garratt-Reed Dr. Adam Papworth Dr. Michal Jarnik Dr. Richard Leapman Dr. Charles Garber Dr. Yasuo Ito Dr. Roseann Csencsits and Dr. Gerd Duscher
} Dear Hildy } I have had good results with neural preservation-- fixing with 4% } formaldehyde (EM grade, no methanol) +.2% glutaraldehyde in PBS. } Dehydration was partial and in a methanol series with progressive } lowering of temperature, } infiltrated in LR white at -20C, and polymerizing at 55C. The series of } methanol was 60% for 15' at 4C, 70% methanol for 60' at -20C, and 80% } methanol for 60' at -20C, then 3 changes of LR white for 60' at -20C. Hope } this will help. This method has given good antigenicity with a wide } variety of tissues. If you need further info, please feel free to contact } me. } Marge } } Margaret Springett } e-mail hukee.margaret-at-mayo.edu } IEM Specialist at Mayo Foundation } 1426 Guggenheim } Rochester, Mn. 55905 }
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
A major development in ion milling is just now being released and may be applicable to your problem. Amorphous damage (as well as implanted Ga fr= om FIB milled samples) can be removed using extremely low energy (100eV - 1000eV) ion milling. Even at Ion energies of 500eV, you can operate at 20uA with a sputtering speed of 2.5 microns per hour. This new technolo= gy provides a means to reduce or eliminate amorphous damage without sacrificing milling rates. Obviously, purchasing a new system may not be= a feasible solution for you, but it is important to know that the technolog= y is available. =
Very useful information on Low Energy Ion Milling can be found in the paper: "Low Angle and Low Energy Ion Beam Etching for TEM Sample Preparation" by Arpad Barna. Proceedings of the Multinational Congress o= n Electron Microscopy Portoroz, Slovenia October 5-8, 1997. =
As I understand that this paper may not be easily accessible to many of you, I can make copies available to you at no charge if you send me your name and address. I also have many other papers on TEM sample preparatio= n which you might find of interest. If you have an interest, I can send yo= u a listing of the available papers.
Two other suggestions for your preparation may be to use the MicroCleave Technique which would eliminate ion milling altogether. The best people = to talk to about this technique are John McCaffrey (john.mccaffrey-at-nrc.ca) a= nd Scott Walck (walck-at-ppg.com).
The other suggestion is to minimize ion milling as much as much possible = by Tripod Polishing your samples into a wedge. You can produce a very thin sample which will require very little if any ion milling. The best conta= ct for this is Ron Anderson at IBM Analytical Services (anderron-at-us.ibm.com)=
The Pathology EM lab here still uses MT1s and MT2s to do thicks and thins. I thought I once saw an upgrade system for MT2s that would upgrade the knife stage to caliper movement and back lighting, and the specimen arc to caliper movement. Does anyone know if such a thing exists? I know the economics of upgrading an MT is a waste of money but I seriously don't think they would spend the additional money for even a used digital system. They are looking for a new tech and I will have to teach them how to section. My Ultracut and MT7 are older but so much nicer to teach with. If the new employee at least had smooth knife and specimen adjustment capabilities it would make learning so much easier. Thanks in advance.
Are you working on polycrystalline Solar Cells? That's what I was doing when I was involved with this material, trying to analyze it with a TEM. We usually had it on thick glass, approx. 3mm thick, with a metal contact between the CIS and the glass.
Let's see if I remember how we prepared that (It's been a few years):
The problem always was the glass, as you suggest in your email. We tried to transfer the CIS to Si before attempting to prepare it for TEM. The way we did that was by mechanical thinnig of the glass as far as possible. The sample was then glued onto Si with the CIS side down. After that, we dissolved the glass in HF. This left us with a much more manageable CIS on Si.
For cross sections, two, sometimes more, of these pieces (thinned) were then glued together and cross sections were prepared. To prevent them from falling apart, the TEM samples were stabilized with a small steel ring (Kim Jones from NREL had a Poster about this stabilizing technique at one of the MSA conferences, I believe). Further preparation then proceeded with mechanical dimpling and ion milling. I always used LN2 ion milling. Without this, artifacts would form, mostly In-rich "droplets". We were able to prepare beautiful samples with this technique, even with a metal backside contact. A 300 KV TEM may have helped, too. We could even do atomic resolution on these samples. That, however, may depend on your sample. if you have very small grains, they may fall out before the sample is done.
For plan views, I cut the TEM sample out of the Si, and thined it mechanically. It was then also stabilized with a steel ring. I then etched a hole into the Si from the backside. That way I was able to prepare free standing CIS films, which could then be ion milled. Perhaps that only worked because I had a metal contact underneath the CIS. It was a bit tricky, because the films sometimes would rip and curl.
I was told by the crystal growers that CIS is not soluble in HF and the comparison samples we prepared this way and the more traditional way never showed any differences.
Since I have not been working with this material for a few years, you should perhaps try to get in touch with Kim Jones at NREL. If you want to, I can get you his email. Please contact me through email.
Hope this helps.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: Albert Romano-Rodriguez[SMTP:romano-at-el.ub.es] } Sent: Wednesday, November 18, 1998 8:56 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM: Preparation of CuInS2 on glass } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The preparation of a concoidal "microchip" of your coating on a glass substrate provides a very thin edged sample suitable by itself as a plan view TEM specimen and it is also an excellent sample for ultramicrotomy to prepare a TEM cross-section. When this thin edged sample is broken to produce a thin and pointed sample it should be treated with an adhesion promoter, embedded in epoxy, cured and then sectioned using a diamond knife. CIS on glass should be a relatively easy material to section by ultramicrotomy.
Phil Swab Advanced Coatings Division/ART
} -----Original Message----- } From: Albert Romano-Rodriguez [SMTP:romano-at-el.ub.es] } Sent: Wednesday, November 18, 1998 10:56 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: TEM: Preparation of CuInS2 on glass } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Dear colleagues, } } We are working on the TEM characterisation of CuInS2 (usually } called CIS) on glass substrates. We are preparing TEM samples out of } these materials (both plan view and cross-section) and we are } encountering problems with the preparation procedure, which are } strong amorphisation of the CIS layer in plan-view and strong damage } (not always amorphisation) in cross-section. } } The preparation procedure we use is the standard preparation } method we use for Si-based materials: For cross-section, we cut } stripes out of the samples, glue them together. Next flat grinding, } dimpling and finally ion milling are used. For plan-views, a piece of } sample is ultrasonic cut and the preparation continues with the flat } grinding, ... and ion milling only from the backside. } } We believe that the problems we have are strongly related to the } glass substrate and to the charging of the glass during ion milling, } resulting in an overheating of the sample. This leads to extremely } poor cross-sections and to even worst plan-view samples (however we } still have some good results from few of these samples). It might be } also that the CIS layer is strongly beam sensitive (any experience } with it?). } } Any help in trying to circumvent these problems will be } extremely helpful and experience in preparing samples of the type } glass substrate-thin layer would also be strongly appreciated. } } By the way our ion milling machines can work at } liquid nitrogen temperature in order to minimise sample heating, if } this helps. } } Thank you in advance for your answers. } } Albert Romano-Rodriguez } Dept. of Electronics } Faculty of Physics } University of Barcelona } c/ Marti i Franques, 1 } E-08028 BARCELONA } Spain } tel: +34-93-402 11 47 } FAX: +34-93-402 11 48 } e-mail: romano-at-el.ub.es }
You may be losing some of the sulfur out of your sample when you ion mill. It is very easy to overheat the glass in the ion mill. You may also be having problems with In. See Cullis and Chew's paper on reactive iodine milling of In containing semiconductors in the MRS vol 115, 1988. You don't say what ion mill that you are using or the conditions. Someone has already suggested tripod polishing and that is a definite possibility, but I don't know how the CuInS2 reacts with water. (Try smelling it while you grind it on a 600 grit sheet of SiC with water as a lubricant. You should also smell the ion mill immediately after ion milling finishes.) Low angle milling might help you tremendously and it would avoid preferential sputtering effects and heating effects. I have not had problems with charging of glass samples but I mill at angles of less than 12 degrees and do quite a bit of it around 5-6 degrees at 5kV.
You could use the small angle cleavage technique with the glass if your films are adherent. The following steps are primarily for XTEM samples but if you read our paper that John McCaffrey sent to you, you will see that you can also pick out some samples that are suitable for plan view.
You may have problems with both water and heating your samples, but I don't know for sure. You can use SACT without water and without heating. Use a superglue for mounting instead of low temperature wax, but you will need to wait very long times to get the samples off of stubs with acetone to avoid heat. Use a viscous slow setting epoxy (super strong types that have about a 1hour working times) for mounting the samples on the grids. These take about a day to cure and again avoid heating the sample. This is explained in our paper. (MRS vol 480, 1997)
Modified steps for glass: 1. Grind the samples to about 75-80 um. on the backside 2. Assuming that you have rectangular samples, use a very coarse SiC paper (180 or 240 grit) and grind in straight lines parallel to the long axis of the sample. This will put your preferred crack directions into the glass, much like semiconductors have a preferred cleavage direction. When you flip the sample over, you will be able to see these scribe lines if your film is thin enough. 3. While the sample is still mounted, scribe the fracture lines at the 15 degree angle on the back side of the sample relative to these scratches, i.e. relative to the long axis of the sample. The distance between lines should be about 5mm. 4. Demount the sample, and break into the strips defined by the scribe lines. 5. Pick the strips of samples up and place on a post-it strip taped to a flat metal block with the sticky side of the post-it up. 6. Use your diamond miniscribe to "cleave" the sample along the scratches that were put there by the grinding with coarse SiC paper. 7. Place the pieces that you "cleave" that are sharp and possible good samples in the sticky corner of your post-it paper for examination, selection, and mounting on the girds for later.
Note: You can use No. 0 or 1 glass cover slips that are pre-ground to suitable thickness for your depositions. Then you can prepare the samples much faster.
South Bay Technology sells the "MicroCleave Kit" that John McCaffrey and myself had a hand in helping them put together. It has all the components that you need to get started doing the technique. With the kit, you really only need a good stereomicroscope.
I have no financial interests with SBT other than a customer, but I do have good friendships with the people that work there.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Albert Romano-Rodriguez To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Dear colleagues,
We are working on the TEM characterisation of CuInS2 (usually called CIS) on glass substrates. We are preparing TEM samples out of these materials (both plan view and cross-section) and we are encountering problems with the preparation procedure, which are strong amorphisation of the CIS layer in plan-view and strong damage (not always amorphisation) in cross-section.
The preparation procedure we use is the standard preparation method we use for Si-based materials: For cross-section, we cut stripes out of the samples, glue them together. Next flat grinding, dimpling and finally ion milling are used. For plan-views, a piece of sample is ultrasonic cut and the preparation continues with the flat grinding, ... and ion milling only from the backside.
We believe that the problems we have are strongly related to the glass substrate and to the charging of the glass during ion milling, resulting in an overheating of the sample. This leads to extremely poor cross-sections and to even worst plan-view samples (however we still have some good results from few of these samples). It might be also that the CIS layer is strongly beam sensitive (any experience with it?).
Any help in trying to circumvent these problems will be extremely helpful and experience in preparing samples of the type glass substrate-thin layer would also be strongly appreciated.
By the way our ion milling machines can work at liquid nitrogen temperature in order to minimise sample heating, if this helps.
Thank you in advance for your answers.
Albert Romano-Rodriguez Dept. of Electronics Faculty of Physics University of Barcelona c/ Marti i Franques, 1 E-08028 BARCELONA Spain tel: +34-93-402 11 47 FAX: +34-93-402 11 48 e-mail: romano-at-el.ub.es
On Wed, 18 Nov 1998, valdemar wrote: } Dear Colleagues, } Could you kindly inform me if there is a forum similar to MSA for Raman Spectroscopy or micro-Raman Spectroscopy? } Berta } berta_m_t-at-hotmail.com
I established and maintain at Williams College a listserv called irusers-l. The listserv is subscribed to by a small number (about 68) scientists around the world who use FT-IR to analyze historic and artistic works. Some of the members, including myself, are interested in the complementary nature of raman, and would benefit from discussion of this technique, in addition to IR.
We're a quiet group, I suppose, but the low volume of discussion on the listserv has caused me to consider opening membership to IR spectroscopists (and possibly Raman spectroscopists) who do not work specifically with historic and artistic works.
My question is, are there members of this listserv who might be interested in participating in a listserv for IR spectroscopy and microspectroscopy or Raman?
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center
Hello photon fans, I need an close loop, contactless system that can check & correct the working distance between objective & glass plate to within 1u. I would appreciate any feed back this group can provide. Please contact me directly if your company has markets this type of product.
I'd welcome feedback on an observation I made this afternoon, while examining a sample from an architectural finish, which included shellac, using epi-fluorescence illumination (details later).
The sample was dispersed in Cargille meltmount 1.662 between a glass slide and cover glass. When illuminated with wide-band ultraviolet illumination (HBO 100 Hg burner and UPlan fluorite objectives) gas began to evolve from the sample, displacing the surrounding mounting medium and leaving small voids within the coating sample. The rate of gas evolution increased with an increase in objective magnification/numerical aperture, and ceased when the vertical illuminator shutter was closed.
I have noted expansion of air pockets in samples before, and have witnessed coating layers melt in a cross-section sample before (both under epi-fluorescence illumination), but have never before witnessed uv-induced gas evolution from a sample. My first thought was a gaseous decomposition product from some unidentified component in the coating.
This is my curiosity of the day, and I'd appreciate hearing from anyone who has observed a similar behavior. Thanks.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center
Dear All, We are recently upgrading our Jeol JXA-8600 microprobe by a new ultra-thin window EDS detector. Does anyone know any solution to prevent or reduce a window contamination in that king of machine? How to improve vacuum in this, rather big, and contaminated microprobe chamber? Is that possible to clean this kind of detector window? Thank you for any advice and solution.
Vitaly Gutkin Electron Probe Laboratory
****************************************** ELECTRON PROBE LABORATORY Hebrew University of Jerusalem Institute of Earth Sciences Givat Ram, Jerusalem 91904 ISRAEL
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Hildy, Have you tried pre-embedding ICC? Fix your tissue with a high PAF-low Glut fixation and then vibratome to produce 30-50=B5m sections. Th= en process these for ICC using very small gold or PAP-DAB for final label.=20 Silver intensify the gold or DAB so it will be visible at both LM and EM levels. After ICC, you can osmicate and embed in any resin. We used to do = this with x-sections of rat brain and other tissues. We could check the sections by light microscopy and then embed those showing label. I would e= mbed between plastic coverslips which were weighted down with metal nuts so the tissue sections were very flat. Then I could cut around the edges of the cover slips, pop one off, and place gelatin capsules (with the solid end cut off) over the area of interest. A drop of resin would be put in the capsule and allowed to partially polymerize before filling the capsule and completing polymerization.
This had the advantage of maximum antibody penetration, permits osmium fixation so you can see the membranes, lets you examine tissue after=
labeling with both light and EM and select areas of maximum interest, and permits serial sectioning if necessary.=20 =20 Pleae contact for more details if desired. Debby -----------=20 Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University =20 1057 Whistler Building West Lafayette, IN 47907-1057
Thanks to all who replied so promptly concerning the transfer of Data from a PC to A TN5500. I have one of two choices; Remove my printer and use that port, or try using a $MI string from library 55 (flextran) to accept data from TN Port 1. I may use the second option for a few reasons. I will be able to simply run a flextran program which transfers input to the PC, which then can control the TN5500 which controls the Cameca MBX Probe( hehe i think i need a flowchart to demonstrate this :) )
Those of you who asked how to recieve data from the TN (sending it to the PC), there are several options. To recieve text based data, i use a command } +DV 1. this changes my output from the printer to the serial port. I use hyperterminal to recieve the data on the PC side. As for images and such, I have two options. VISTA, a program supplied with our probe, is supposed to be an imaging program with outputs to the PC (I have never used this route though). We also purchased a GW electronics image capture board (slow scan analog to digital). I understand this board and the accompanying software is quite expensive.
Again thank you for your help Ted Claypool Engineer Scientist / EPMA RJ Lee Group
{!doctype html public "-//w3c//dtd html 4.0 transitional//en"} {html} Thanks to all who replied so promptly concerning the transfer of Data from a PC to A TN5500. {br} I have one of two choices; Remove my printer and use that port, or try using a $MI string from library 55 (flextran) to accept data from TN Port 1. I may use the second option for a few reasons. I will be able to simply run a flextran program which transfers input to the PC, which then can control the TN5500 which controls the Cameca MBX Probe( hehe i think i need a flowchart to demonstrate this :) ) {p} Those of you who asked how to recieve data from the TN (sending it to the PC), there are several options. To recieve text based data, i use a command } +DV 1. this changes my output from the printer to the serial port. I use hyperterminal to recieve the data on the PC side. {br} As for images and such, I have two options. VISTA, a program supplied with our probe, is supposed to be an imaging program with outputs to the PC (I have never used this route though). We also purchased a GW electronics image capture board (slow scan analog to digital). I understand this board and the accompanying software is quite expensive. {p} Again thank you for your help {br} Ted Claypool {br} Engineer Scientist / EPMA {br} {a href="http://www.rjlg.com"} RJ Lee Group {/a} {br} {br} {/html}
If I were you I would look at the Sem-Clean system from XEI Scientific. http://www.msa.microscopy.com/sm/xei
Their system bleeds nitrogen through the system to carry contaminants out of microscope, and it might be just the thing to clean up your probe.
best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
Dear All, We are recently upgrading our Jeol JXA-8600 microprobe by a new ultra-thin window EDS detector. Does anyone know any solution to prevent or reduce a window contamination in that king of machine? How to improve vacuum in this, rather big, and contaminated microprobe chamber? Is that possible to clean this kind of detector window? Thank you for any advice and solution.
Vitaly Gutkin Electron Probe Laboratory
****************************************** ELECTRON PROBE LABORATORY Hebrew University of Jerusalem Institute of Earth Sciences Givat Ram, Jerusalem 91904 ISRAEL
I had a chance to look at the cells I processed using HMDS and here's what I saw... They looked fine. I only went up to about 2,000X but they didn't look any different from the two samples that I ran up in the CPD. I used the slower infiltration of HMDS:
3:1 ETOH : HMDS 15 min. 1:1 E : H 15 min. 1:3 E : H 15 min. 100% HMDS 3 X 15 min.
Then I just put the tops to the dishes slightly askew and then let nature take it's course. I let them evaporate overnight in the hood. Then I put the coverslips onto stubs and sputter coated.
The one bad thing that happened was that some of the cells were grown in chamber slides and the HMDS ended up degrading the silicon seal enough for the stuff to leak out. The cells didn't look like they were too upset by this, though.
Whether this works for all cell lines, I don't know but I did this on loosely adherent tumor cells and lymphocytes.
Thanks to all who gave me suggestions as to what to try. Y'all are a truly great resource and I get lots of useful tips from this list.
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Hello collegues; Is there a forum (like this Microscopy one) for the X-ray phototelectron spectroscopy, or Auger, SIMS, ISS, etc.? If so, please let me know. Thanks. Sincerely yours Ram Srinivasan University of Kentucky
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Responding to the message of {3651F0D1.6C4F-at-botanica.ufrgs.br} } from Rinaldo Pires dos Santos {rinaldop-at-botanica.ufrgs.br} : } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear colleagues } } } } How to improve the contrast in plant tissue included in Spurr resin? The } } double contrast with Uranyl acetate in ethanol solution and Lead citrate } } is not good for me. There is some special procedure for to solve this } } problem? With the ultra-low viscosity resin, the problem is worst (lower } } contrast than the Spurr resin). } } Thank's in advance. } } } } M.Sc. Rinaldo Pires dos Santos } } Dept. of Botany - UFRGS } } Porto Alegre - RS - Brazil } } What kind of plant tissues? } } 1. Try a 50:50 mixture of Spurrs/Embed812 resins. Mix up resins seperately } without their accelerators (DMAE & DMP-30). You can store them in the freezer. } Then mix 50:50 just before use, mix in required amounts of the two accelerators, } infiltrate and cure. You should get the better sectioning and staining } properties of Embed812 plus the low viscosity of Spurrs. } } 2. Switch to Embed 812 100% and extend infiltration times. } } 3. What lead stain are you using? In my experience, Reynold's lead citrate } doesn't seem to work too well with Spurrs. I use Sato's triple lead stain, } preceeded by 3% UA, and get very good staining on plant and bacterial samples } embedded in Embed812. } } Good luck, } } Gib } } } } Gib Ahlstrand } Electron Optical Facility, University of Minnesota, Dept. Plant Pathology } 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 } 612-625-9728 FAX, giba-at-puccini.crl.umn.edu } } } Dear Gib,
You got a problem! Please remember the following: Perfect infiltration (in your case the low viscosity) nixes contrast and immunocytochemistry attempts. You will not get perfect infiltration with epoxides, but then there are other problems. So, first, try this. 1. Use 1% Tannic Acid in cac buffer ,pH6.9, after glutaraldehyde fixation
2. Leave the prep in osmium overnight in the refrigerator.
3. Do an enbloc UA (3% maleate buffer, not phosphate or cac) for 90 minutes in the refrigerator
4. Add p-phenylenediamine to your 70% alcohol mixture - try 1% for 15min
5. Then look at some of the sections before staining in the TEM. Do not attempt to poststain those sections. See what you get.
6. Poststain with both UA and Reynolds. Reynolds yields more gray tones than Sato's.
7. If this does not work, then you must go to a very liquid epoxide, perhaps even a formulation with a dilutent in it.
Let me know if the above works.
Bye,
Hildy
P.S. Cut thicker sections. Use a smaller aperture! "Overexpose" (set up the density on your TEM) your negatives. Print on #1 and # 2 paper only. If you have to print on #3, your negative is not dense enough and you are loosing information. You would be amazed how diddling with the scope settings help! We drive this to extremes in cases with immunocytochem where we cannot use osmium UA, etc.
Note: The above sequence of TA, Osmium, UA, Ppd, UA, Pb, is a "cascade of mordanting agents" One acts as a mordant for the next all along.
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A single crystal of this material was jet electropolished with good success. A similar electrolyte may work for the CuInS2 material Mr. Rodriguez has. A south Bay 550 B single jet polisher was used with the following electrolyte: 60 ml. perchloric acid 460 ml. ethyl alcohol 280 ml. n-butyl alcohol 100 ml. butyl cellosolve 150 ml. acetic acid
Conditions: -20 degrees centigrade, 80 volts, 12 mA. current on a 1.5 mm. diameter exposed area. (One surface at a time). Lacquer protected the polished initial dimple during thinninging to automatic optical termination of the process. Some residue specks were seen on the foil, but more tests with the proportion of the ingredients might reduce that problem.
id {W04KZ8CD} ; Thu, 19 Nov 1998 16:47:38 -0500 Message-ID: {2D0F795D02A5D1118E7700A0C99B3AA23CC5A4-at-CHD64SMAIL01} To: microscopy-at-Sparc5.Microscopy.Com
*if replying please reply to this sender as he is "offlist".*
Hello Microscopy Experts,
At the suggestion of one of your colleagues, I would like to pose my inquiry to your list. I am not a subscriber, so I am sending this "off list" (hope you don't mind). If you wish to reply please reply to my email address {tim_wallace-at-doh.state.fl.us}.
Question 1. Are reference microphotographs available for natural fibers (cotton or cellulose for example)? I am trying to find a reference or photographic atlas to help me when looking in a light compound microscope at house dust samples. Can anyone suggest a web-site or a free/low cost resource with reference photographs or images that would be useful?
Question 2. Is there a lab in the United States that microscopically examines household dust, characterizing and identifying the contents (fibers, particles, hairs, globs etc.)?
I am Environmental Specialist working for a Local Health Department investigating a health complaint/concern from a resident. The resident is worried that the excessive dust problem, may pose a threat to her health. She really wants to know what is in the dust and where is it coming from. Any information would be appreciated. Thanks and best regards,
Tim tim_wallace-at-doh.state.fl.us
________________________________________________ Tim Wallace Environmental Specialist II Volusia County Health Department Environmental Health Division Indoor Air Assistance Lead Monitoring Programs 501 S. Clyde Morris Blvd. Daytona Beach, Florida 32114 USA phone: 904.947.3484 fax: 904.947.3485 http://www.state.fl.us/cf_web/D12/cphu/ehprgms/iaq.html +++++++++++++++++++++++++++++++++++++
XEI Scientific does make a cleaning system that can do what you need. I make a SEM-CLEAN nitrogen purge system for a JEOL 8600 probe, and I would be pleased to build another for you. Give me your mailing address and I will send you complete information.
Ronald Vane XEI Scientific 3124 Wessex Way Redwood City, CA 94061 (415) 369-0133
Vitaly Gutkin wrote: } } Dear All, } We are recently upgrading our Jeol JXA-8600 microprobe by a new } ultra-thin window EDS detector. } Does anyone know any solution to prevent or reduce a window } contamination in that king of machine? } How to improve vacuum in this, rather big, and contaminated microprobe } chamber? } Is that possible to clean this kind of detector window? } Thank you for any advice and solution. } } Vitaly Gutkin } Electron Probe Laboratory } } ****************************************** } ELECTRON PROBE LABORATORY } Hebrew University of Jerusalem } Institute of Earth Sciences } Givat Ram, Jerusalem 91904 ISRAEL } } VITALY GUTKIN } } Phone: 972-2-6585897 Fax: 972-2-5662581 } mailto:vit-at-cc.huji.ac.il } http://earth.es.huji.ac.il/e-prob/prob.html } ******************************************
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Dear Hildy } } I have had good results with neural preservation-- fixing with 4% } } formaldehyde (EM grade, no methanol) +.2% glutaraldehyde in PBS. } } Dehydration was partial and in a methanol series with progressive } } lowering of temperature, } } infiltrated in LR white at -20C, and polymerizing at 55C. The series of } } methanol was 60% for 15' at 4C, 70% methanol for 60' at -20C, and 80% } } methanol for 60' at -20C, then 3 changes of LR white for 60' at -20C. Hope } } this will help. This method has given good antigenicity with a wide } } variety of tissues. If you need further info, please feel free to contact } } me. } } Marge } } } } Margaret Springett } } e-mail hukee.margaret-at-mayo.edu } } IEM Specialist at Mayo Foundation } } 1426 Guggenheim } } Rochester, Mn. 55905 } } } } Margaret Springett } e-mail hukee.margaret-at-mayo.edu } IEM Specialist at Mayo Foundation } 1426 Guggenheim } Rochester, Mn. 55905 } } } } Hi,
The above is an excellent protocol. We have found it to visualize antigens which absolutely will not allow themselves to be detected any other way. We are getting great label. Our problem is that we can only use 0.25% GA (I hate this antigen) and the structure which we need to show up (vesicles) do not seem to preserve enough for us to call them vesicles in a publication. I recommend your protocol highly to anyone who has trouble with epoxides in immuno. (Our preservation in expoxides was quite good even without osmium, but vesicles tend to be ephemeral anyway, and the poor preservative ability of the LRs seems to have put them over the edge). Thanks, Hildy
Vitaly Gutkin wrote: {snip} } How to improve vacuum in this, rather big, and contaminated microprobe } chamber? } Is that possible to clean this kind of detector window? } Thank you for any advice and solution. } } Vitaly Gutkin } Electron Probe Laboratory } } ****************************************** } ELECTRON PROBE LABORATORY } Hebrew University of Jerusalem } Institute of Earth Sciences } Givat Ram, Jerusalem 91904 ISRAEL } } VITALY GUTKIN } } Phone: 972-2-6585897 Fax: 972-2-5662581 } mailto:vit-at-cc.huji.ac.il } http://earth.es.huji.ac.il/e-prob/prob.html } ******************************************
First, of course, clean the chamber well.... Perhaps water/detergent, alcohol, acetone, etc... will depend on cantaminants.
The windows can be cleaned by carefully dripping solvent across the window. There has been a thread about this recently on the Microscopy Society of America Listserver.
Keeping it clean:
Not familiar with specifics of your JEOL, but have been running a Be/UTW/Open Window EDS detector for 15 years and never had an oil problem. Only a few times have I had to warm the detector to clear the (cryo pumped) crystal. Most of this problem I attribut to P-10 gas (argon/methane) from a less-than-perfect WDS thin window leak. The key may not be cheap, depending on your configuration. My SEM is an ETEC which has staged vacuum pumping. The chamber is rough pumped to 100 microns. At this point vacuum valving places a Magnetic levitation bearing turbo pump ahead of the rough pump. ..Very clean vacuum. The only improvement would be to use an oil-less rough pump, but I haven't had the funding.
Regards, Woody
--------------------------- -- ------------------ de Woody, WB4QXE -------------------
- Work: SEM/EDS/WDS - Materials Research & Failure Analysis also, electronics and instrumentation.
- Home: Ham radio "homebrewing", computers , shade tree mechanic.
- www site: Scanning Electron images and Ham Radio Homebrewing stuff. http://www.geocities.com/capecanaveral/3722
The only forum I know is for SIMS. The web address is listed below. You can also subscribe to a list server from the site. http://www.simsworkshop.org/default.nclk
Hope this helps, Ed Hirsch
At 11:22 AM 11/19/98 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am still working on my project to teach histology to highschool students. People on this list have been very helpful in getting me a microtome and prepared slides. I am now working on the chemicals & supplies I need. Going over an on-line catalog I discovered that there are much safer substitutes for the chemicals used in my 1962 edition of "Animal Tissue Techniques" by Humason. However, it was not clear to me what safer solvent is used to substitute for toluene in the paraffin infiltration stage. Can anyone suggest an up to date reference book which discusses the advantages and disadvantages of the new substitutes. It appears that a lot has changed since I worked in a medical lab 30 years ago. Thanks.
The best source for particle information is from McCrone Accessories. It is called the Particle Atlas and may be available through a local university science library. Commerically, it is available on CD Rom, for about $1100. It is a really great resource (we've used it with several clients and classes). The McCrone Institute is also a good lab for this type of thing. Closer to your area is a high quality spin-off, run by Skip Palenik. The contact numbers for both MAC and the Institute are below. I don't have current info for Skip but imagine that you can find him on the Web.
} Dear All, } We are recently upgrading our Jeol JXA-8600 microprobe by a new } ultra-thin window EDS detector. } Does anyone know any solution to prevent or reduce a window } contamination in that king of machine? } How to improve vacuum in this, rather big, and contaminated microprobe } chamber? } Is that possible to clean this kind of detector window? } Thank you for any advice and solution. } } Vitaly Gutkin } Electron Probe Laboratory } } } ****************************************** } ELECTRON PROBE LABORATORY } Hebrew University of Jerusalem } Institute of Earth Sciences } Givat Ram, Jerusalem 91904 ISRAEL } } VITALY GUTKIN } } Phone: 972-2-6585897 Fax: 972-2-5662581 } mailto:vit-at-cc.huji.ac.il } http://earth.es.huji.ac.il/e-prob/prob.html } ****************************************** } Dear Vitaly! The radical solution is to replace diffusion pump oil by Santovac-5. I made it some years ago on the predecessor of your microanalyzer - JEOL Superprobe JXA-733. In result the vacuum was improved everywhere on the order. Contamination of sample under beam have decreased considerably (analysis of light elements has become more accurate). Besides I have become to clean the column 1 time per 4 years (before it was 1 time per 3 months). Precautions: 1. Santovac-5 has more viscosity therefore probably it will be necessary to increase slightly the power of diffusion pump heater, for example by autotransformer. 2. To notice effect it is necessary to clean well the column and vacuum system from old oil (most labour-consuming procedure). I would like to advise also to install a trap for forepump oil vapours and the liquid nitrogen trap for diffusion pump and plate above sample holder. Though from conference it is known all EDS manufacturers make windows in one place, it seems better to request of the manufacturer for recommendations on window cleaning. For example Oxford Instruments has this information. Regards.
Victor Sidorenko, ANTRON Co. Ltd., scientific service, Moscow, Russia antron-at-space.ru
The only forum I know is for SIMS. The web address is listed below. You can also subscribe to the SIMS list server from the site. http://www.simsworkshop.org/default.nclk
Hope this helps, Ed Hirsch
At 11:22 AM 11/19/98 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } In regards Beth Richardson's search for a new blade for the Dahle paper cutter. You can get one. I have had my Dahle for over 10 years and replaced the cutting blade about a year ago. I called my local photography supply house and they ordered a new blade for me. It was easy to put in and I've been using it ever since. It's cheaper than buying a new cutter. God luck. } JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94022
One of the best histology books out today is=20 Histotechnology, A Self-Instructional Text=20 by Freida L. Carson
ISBN 0-89189-306-7
You can be ordered the book through the American=20 Society of Clinical Pathologists
George Lawton Chief Electron Microscopist Microscopy and Imaging Center UT Southwestern Medical Center at Dallas Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
Why is Hepes buffer not used in the primary fixation with glutaraldehyde for mammalian tissues instead of the phosphate and cacodylate buffers? Anyone know the biochemical reasons?
Apparently in my last posting concerning Low Energy Ion Milling, Tripod Polishing and the MicroCleave Technique my disclaimer and signature line did not appear. While it did appear on my copy of the message, for some reason it did not appear on the copy of the message that I saw on the Listserver. I am sure that the people on the list know that we produce these products, but I thought I should make it clear by reiterating my disclaimer here. So here goes:
NOTE: We do sell the IV3 Ion Milling System with LEGs (Low Energy Guns) a= s well as the MicroCleave Kit and the Tripod Polisher so I do have a vested=
For some of us, HEPES is the buffer used for primary fixation with aldehydes. I find it works fine. PIPES would be another good choice (it has a slightly lower pKa so would have greater buffering capacity that HEPES if both started at 7.4 and the solution had a tendency to acidify (like aldehyde fixatives do).
} Hi again, } } Why is Hepes buffer not used in the primary fixation with glutaraldehyde } for mammalian tissues instead of the phosphate and cacodylate buffers? } Anyone know the biochemical reasons? } } Thanks, } Hildy
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Skip Palenik is on the Web at palenik-at-aol.com and is an excellent resource. His Company Name is Microtrace in Elgin, IL Phone: 847-742-9909
I have no financial interest in this recommendation.
Tom Kremer Analytical Science & Technology Kimberly-Clark 920-721-4583 e-mail: tkremer-at-kcc.com
} ---------- } From: Barbara Foster[SMTP:mme-at-map.com] } Sent: Friday, November 20, 1998 9:26 AM } To: Tim_Wallace-at-doh.state.fl.us-at-sparc5.microscopy.com; } microscopy-at-sparc5.microscopy.com } Subject: Re: Microscopic House Dust Characterization & ID } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Tim, } } The best source for particle information is from McCrone Accessories. It } is called the Particle Atlas and may be available through a local } university science library. Commerically, it is available on CD Rom, for } about $1100. It is a really great resource (we've used it with several } clients and classes). The McCrone Institute is also a good lab for this } type of thing. Closer to your area is a high quality spin-off, run by } Skip Palenik. The contact numbers for both MAC and the Institute are } below. I don't have current info for Skip but imagine that you can find } him on the Web. } } } McCrone Accessories: 800-622-8122 } McCrone Institute: 312-842-7100 } } Best of luck, } Barbara Foster } Consortium President } Microscopy/Microscopy Education ..Educating microscopists for greater } productivity. } } 125 Paridon Street Suite 102 Springfield, MA 01118 } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } Visit our web site {http://www.MME-Microscopy.com/education} } ****************************************************** } MME is America's first national consortium dedicated to } customized on-site training in all areas of } microscopy, sample preparation, and image analysis. } } } } At 04:45 PM 11/19/98 -0500, } Tim_Wallace-at-doh.state.fl.us"-at-Sparc5.Microscopy.Com wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } *if replying please reply to this sender as he is "offlist".* } } } } Hello Microscopy Experts, } } } } At the suggestion of one of your colleagues, I would like to pose my } inquiry } } to your list. I am not a subscriber, so I am sending this "off list" } (hope } } you don't mind). If you wish to reply please reply to my email address } } {tim_wallace-at-doh.state.fl.us}. } } } } Question 1. Are reference microphotographs available for natural fibers } } (cotton or cellulose for example)? I am trying to find a reference or } } photographic atlas to help me when looking in a light compound microscope } at } } house dust samples. Can anyone suggest a web-site or a free/low cost } } resource with reference photographs or images that would be useful? } } } } Question 2. Is there a lab in the United States that microscopically } } examines household dust, characterizing and identifying the contents } } (fibers, particles, hairs, globs etc.)? } } } } I am Environmental Specialist working for a Local Health Department } } investigating a health complaint/concern from a resident. The resident } is } } worried that the excessive dust problem, may pose a threat to her health. } } } She really wants to know what is in the dust and where is it coming from. } } } Any information would be appreciated. Thanks and best regards, } } } } Tim } } tim_wallace-at-doh.state.fl.us } } } } ________________________________________________ } } Tim Wallace } } Environmental Specialist II } } Volusia County Health Department } } Environmental Health Division } } Indoor Air Assistance Lead Monitoring Programs } } 501 S. Clyde Morris Blvd. Daytona Beach, Florida 32114 USA } } phone: 904.947.3484 fax: 904.947.3485 } } http://www.state.fl.us/cf_web/D12/cphu/ehprgms/iaq.html } } +++++++++++++++++++++++++++++++++++++ } } } } } } } } } } }
In a message dated 98-11-20 17:07:30 EST, hcrowley-at-du.edu writes:
{ { Does anyone know if enbloc uranyl acetate staining interferes with UV Polymerization of LR White or LR Gold at -20C? Has anyone done it? Thanks, } }
Hi Hildy,
I don't know about LR White and Gold, but en block stains can sure cause problems with Lowicryl K4M, which is also polymerized by UV. I would guess that you may encounter some problems, probably in the centers of the specimens.
Cheers, Bob **************************************** Robert (Bob) Chiovetti, Ph.D. Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems *****************************************
The following showed up on the Sci.Techniques.Microscopy Usenet Newsgroup. Someone on this list may be able to help him. If so, please respond directly to him at:
emans-at-bio1.rwth-aachen.de
Please don't respond to this message you are reading now, since I have only copied the text into this message.
Thanks.
Bob Chiovetti
***************************************
I keep seeing posts mentioning the McCrone Research Center on this list server. I have never seen it mentioned that they have a web page. So for those interested, they do. It is:
HILDEGARD CROWLEY wrote: } } Hi, } } Does anyone know if enbloc uranyl acetate staining interferes with UV } Polymerization of LR White or LR Gold at -20C? } Has anyone done it? } Thanks, } Hildy
"Use of uranyl acetate en bloc to improve tissue preservation and labeling for post-embedding immunoelectron microscopy" Erikson, P.A. et al. 1987. J. Elec. Micros. Tech. 5:303-314.
They stained en bloc with UA before embedding in LR White or Lowikryl K4M. Of course, the antigen in question might make a difference.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
We are planning to rebuild the optical diffractometer we use to assess the quality of our TEM micrographs. Anyone have a favorite design or modification for an optical diffractometer (and perhaps an associated reference)? We are looking to build a vertical set up similar to the one in:
We'd also like to add a CCD camera with a high quality thermal printer.
Any recommendations?
Thanks,
- Dennis
------------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
To Harris Reavin, The evolution of clearing solvents is interesting. Toluene (actually chloroform) works best, but is a no-no because of carcinogenicity and environmental (disposal) problems. Xylene has been the solvent of choice for both clearing and dewaxing, but again, health effects and disposal are major concerns. Limonene products came on the scene as the "magic" xylene substitute, boasting the fact(?) that they could be dumped down the drain. Well, in my experience they don't perform well, our local sewer authorities don't want them in the system and you can smell them out in the parking lot! Aliphatic hydrocarbons, such as certain products from Richard-Allen Scientific 800-522-7270 and ANATECH 800-ANATECH have a proven track record as xylene substitutes. Check with these companies, you might persuade them to send you some samples...and they both are very knowledgeable and helpful and offer all kinds of expert advice. In my opinion, you'll get good clearing with aliphatic hydrocarbons, but you will have to do your whole processing procedure under a fume hood. Remember gloves, goggles and apron...be safe! Good luck, Bob Santoianni Emory University Hospital Atlanta, GA robert_santoianni-at-emory.org
Grateful thanks for the helpful advises and comments to anybody, who response my questions about JXA-8600 microprobe cleaning.
Yours sincerely,
Vitaly Gutkin Electron Probe Laboratory
-- ****************************************** ELECTRON PROBE LABORATORY Hebrew University of Jerusalem Institute of Earth Sciences Givat Ram, Jerusalem 91904 ISRAEL
The E.M. Unit in Triniy College Dublin have a secondhand Emscope SP2000A Cryogenic preparation system for sale. This is a fully working system with a gas cooled stage, rather than the usual braid system, to give a rapid response to cooling and heating requirements. The asking price is =A35,000 sterling with the purchaser arranging carriage and insurance. =46or full details please contact me off list.
David John, Manager, Electron Microscope Unit, Trinity College, Dublin 2, Ireland. tel.no. (353) - 1 - 6081559 e-mail - djohn-at-mail.tcd.ie
Last week during the thread on CuInS2 on glass, someone from Raytheon somewhere in Texas sent me a personal email asking for a copy of the Small-Angle Cleavage Technique video tape. We routinely mail these out like a paper reprint, but I inadvertantly trashed his message and lost his mailing address. So, will that person from Raytheon (or anyone else who wants the video) please send me another message, and I'll send him a copy of the tape. My apologies to everyone else for filling up your email message box.
Cheers John
John P. McCaffrey Institute for Microstructural Sciences National Research Council of Canada M-50 Montreal Rd. Ottawa, Ontario K1A 0R6 Canada
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RE} McCrone Res. Center on web 11/23/98 8:55 AM Thank you Azriel for the mention of the McCrone web site.
The "home" web address is:
http://www.mcri.org
} From there you may go to any of our pages and find out about us, our cour= ses, our expertise, mission, etc.
Thanks.
John D. Shane Director of Research McCrone Research Inst. 2820 South Michigan Ave. Chicago, IL 60616 312.842.7100
--------------------------------------
I keep seeing posts mentioning the McCrone Research Center on this list server. I have never seen it mentioned that they have a web page. So for those interested, they do. It is:
Does anyone know how the get information on durability and storage of B&W prints from the thermal printer. We use Seikosha VP-3500 printer, but we were not able to get any information from the manufacturer.
I am looking for a used color monitor for a Kevex Delta 1 EDX System. It is a model 38-DO5IMA-UU free standing monitor with 5 RCA plugs exiting the rear.
If anyone can help, please contact me at 303-689-2224 or e-mail franklin-at-idcomm.com...
Hi all. Yes this is real boring technical stuff but we hope it helps somebody. A while ago we asked if any one else was having problems with some of = the older ISI SEM's regularly blowing the transistors in the box on top = of the HT tank. We had quite a response from many people having the same = problem but not may solutions to the problem that seemed to work.
We think we have been able to identify the problem and also solve the = problem.
The cct inside the tank on the bias and HT side that has the 4700pF and = 2200pF caps on it, is to filter out the back emf from the high voltage = side of the tank. We found that these same caps go soft and so leak that = voltage back up into the driver transistors on top of the tank. This is = what causes them to blow. It is quite simple to monitor this. If you = measure the collectors of these transistors you should measure waveforms = with a peak voltage not exceeding 400v on any of the transistors. ( the = actual voltage will vary with bias settings, filament current and HV.) = Should you find that the voltage is close to this voltage or over, then = replace the relevant caps in the tank ( C20, C21, C4, C5 etc. 4700pF = 2.5Kv and 2200pF 4Kv and 0.1uF 630v) This will solve the problem.
Cheers Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
Fellow microscopists, I am setting up a darkroom for my electron microscope laboratory recently and looking up a suitable enlarger. I would appreciate your suggestions and comments on what brand/ specifications I need for my TEM, SEM and optical microscopes. Thanks Ren-Jye
We have a Seikosha VP-3500 and I can give you some input.
These prints will degrade very rapidly if left out in the lab where they are exposed to fluorescent lights. You can see degradation beginning to occur in a week or so.
Best bet is to store them in a dark cabinet, this will help slow the degradation process. I have pictures that are several years old stored that way.
I wouldn't count on them for long term storage of critical information. we went to a relatively cheap digital image capture system (Snappy Image capture card on the NTSC outlet of our AMRAY 1830) and are well pleased with the results. We typically take both thermal prints and a digital image on all images.
John Giles Principal Materials Engineer Honeywell Space Systems
} Hi everyone,
} Does anyone know how the get information on durability } and storage of B&W prints from the thermal printer. We } use Seikosha VP-3500 printer, but we were not able to } get any information from the manufacturer.
On Mon, 23 Nov 1998, Craig Franklin wrote: } I am looking for a used color monitor for a Kevex Delta 1 EDX System. It is } a model 38-DO5IMA-UU free standing monitor with 5 RCA plugs exiting the } rear.
If those plugs are labelled R,G,B,H,V, it is probably a standard RGB monitor and there are many replacements from many manufacturers. If it is an older machine, the bandwidth is probably not great making replacement relatively easy. Surely, someone in the lab/department has a computer monitor with RGB inputs you could try out.
: On Mon, 23 Nov 1998, Craig Franklin wrote: : } I am looking for a used color monitor for a Kevex Delta 1 EDX System. It is : } a model 38-DO5IMA-UU free standing monitor with 5 RCA plugs exiting the : } rear. : : If those plugs are labelled R,G,B,H,V, it is probably a : standard RGB monitor and there are many replacements from : many manufacturers. If it is an older machine, the : bandwidth is probably not great making replacement : relatively easy. Surely, someone in the lab/department has : a computer monitor with RGB inputs you could try out. :
The monitor you have was most likely made by Electrohome.
A standard RGB monitor will work, but only if it can sync at low H sync rates. Our 8000 puts out an H sync of 17 Hz and a V sync of 57 Hz.
Carl
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA -------------------------------- Voice: (734) 936-1550 FAX: (734) 763-4690 E-mail: chender-at-umich.edu --------------------------------
Sorry for the delay in summarizing all the feedback ... I figured the best way would be just to cut-and-paste everyone's advise onto one email because I could miss certain points.
All advice has been a great help.
Thank you, Ad
+++++++++++
} From:Yasuo Ito
Hi, I got your message through microscopy list server. Yes, i think you have a tough assignment since sulphur L-edge(I presume that you are looking at these edges) is at about 165 eV. My guess is that (1) the thickness of your specimen may be too thick (2) interefered by any other edges, and/or (2) sulphur might be removed due to the electron-beam damage.
For (1), if the speciment is too thick, the plasmon loss peak becomes to big and swamp the sulphur peak. How thick is your specimen? I mean how thick in terms of inelastic mean free path. You can measure this by taking low loss spectrum of the are of interest. I guess, the thickness would be less than a half of that.
For (2)and (3), this depends on what is the matrix of your specimen.
For (3), you can monitor any change of the mass thickness by monitoring plasmon peak, as you may know. Or at least you may be observe by images. Have you seen any indication of the beam damage? Do you have EDS? you may be able to check by the EDS for the existence of sulphur in your specimen. If the beam damage is the case, you may have to reduce the dose into the area by reducing beam current or acquisition time.
So far this is what I can think of at hand. I hope these would give you some idea (I guess you may have already thought about these possibility, though.)
Please don't hesitate to contact me for any further questions. I have been dealing with beam sensitive materials for a while.
Cheers,
Yasuo
++++++++++++
} From:Larry Thomas
Advice is cheap, so I'm told, but here goes.
I would take not being able to see the sulfur edge as a bad sign. Maybe there is really no S in your sample. Are you sure the 'sulfur ' peak you saw by EDS wasn't actually a Mo peak or Pb peak from the sample or from a microscope artifact. It's not unusual to get a small Mo x-ray contribution from scattering off the condenser apertures in the TEM. You say the sulfur concentration in the sample "is not that much." How much is that? The detection limit in EELS might be as much as several percent. If the signal is being hidden by the huge background in EELS, the most sensitive way I know of to detect it is by using 2nd difference spectra. Difference collection modes remove background much more effectively than power law subtraction. There is an optimum collection angle in EELS, so you might try different collection apertures. If you can't detect the sulfur in difference spectra with enough counts for the detection limits you need, don't expect anything useful from power law correction, and especially not from mapping with very limited counts from individual analysis points.
I would also ask myself the question "If I'm getting easily detectable sulfur by EDS analysis, why bother mapping with EELS?"
Anyway, that's my two-cents worth.
Larry Thomas Washington State University
+++++++++++++++
} From:Gerd Duscher
I am working mostly with a VG HB501 with Gatan PEELS for several years. So in principle I have the same sensitivity for sulphure as you.
I think your experimental conditions are fine!
I had a similar problem with Silicon, which I solved using two different approaches.
First I would collect two spectra with good counting statistics in the sulphur and in the low loss region. Splice them and do the single scattering deconvolution using the LOG-ratio method in EL/P. This ensures that you get always the same background before the edge and you cancel out thickness effects.
Secondly, if you have regions without sulfur and without, you can use the spatial difference technique: author = {H. M\"ullejans and J. Bruley}, title = {Improvements in Detection Sensitivity by Spatial--Difference Electron Energy--Loss Spectroscopy at Interfaces in Ceramics}, journal = {Ultramicroscopy}, year = 1994, volume = 53, number = 4, pages = {351-360},
You need two spectra which are taken under the same conditions and at locations with the same thickness. Then you use the one without for an improved background subtraction.
I have just noticed your message about sulfur mapping. I wonder if you have seen the following publication?
Spatial distributions of sulfur-rich proteins in cornifying epithelia. Leapman, RD, Jarnik, M, Steven, AC. J Struct Biol 1997; 120: 168-179.
I shall be pleased to send you a reprint if you give me your mailing address.
...Richard Leapman
_____________________________ National Institues of Health Bioengineering & Physical Science Program, ORS Supramolecular Structure & Function Resource Bldg. 13, Rm. 3N17 Bethesda, MD 20892-5766 tel: (301) 496-2599 fax: (301) 496-6608 e-mail: leapman-at-helix.nih.gov
web reference: http://www.nih.gov/od/ors/beps/ssfr/ _____________________________
+++++++++++++++
} From:Charles A. Garber
This might be a dumb kind of comment, so forgive me if you think it is, but one more than a few instances, people have found Mo in their spectra coming from Mo apertures. So you might want to check and see if you are using Mo apertures and if yes, you might want to change them to apertures of some other composition (for example, Pt) and see if your Mo peaks are still there or not.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################
++++++++++++++++++++
} From: Michal Jarnik
Ad,
we recently published an article in the Journal of Structural Biology (Leapman, R. D., Jarnik, M. and Steven, A. C. (1997). Spatial distributions of sulfur-rich proteins in cornifying epithelia. J. Struct. Biol., 120, 168-179). If you send me your mailing address I will be happy to get you a copy.
Regards,
Michal -- Michal Jarnik, Ph.D. Fox Chase Cancer Center Electron Microscopy Facility 7701 Burholme Ave. Philadelphia PA 19111 Tel. 215-728-5675 Fax 215-728-2412
++++++++++++++++++
} From: Haswell Malcolm
I believe that one of the other listers has already mentioned molybdenum peaks. This has been a chronic problem on our Hitachi H7000 because of the Mo in the movable objective apertures.
We have had problems with EDX, not EELS, but I would expect that the geometry may be worse in EELS because your detector might more directly "see" objective apertures and so increase the chance of picking up Mo. If your system will pick up higher energies of Mo it would certainly be worth doing a quick check.
Incidentally I received a lot of advice about our molybdenum problem some time ago and I can't remember if I thanked everyone - so thanks, just in case. Unfortunately all we ever managed to do was minimize the effect, unless we removed the aperture rod which produced its own problems.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
++++++++++++++++++++++
} From:Laryy Thomas
Gerd's suggested method for removing background is technically the best although it involves a bit more work than difference spectrum collection.
You haven't said what S concentration you're actually looking for. What is it?
To look for hard-to-find sulfur in EELS, I'd find the S-containing area with EDS before trying EELS. In EDS or EELS, use an area collection mode such as selected-area diffraction rather than a focused probe mode to minimize beam-damaging the sample. A 50 nm sample thickness should be fine. The broad type of edge sulfur has at ~165 eV is easy to hide in background. At that energy loss, my experience is a power law gives very poor fits. If you have the Gatan DigiPEELS system, has anyone mentioned the correction for point spread in the detection system? It can be important.
As to the comment about preferring EELS mapping over EDS mapping because of much longer necessary acquisition times in EDS, I don't see why this should be so. Certainly you should optimize the acquisition conditions in EDS (you can use a short processing time and high beam current to optimize counting rates), but you have to do essentially the same thing in EELS anyway. The question to ask yourself is "What detection limit do I need for this job and therefore, what count rate do I need to get good statistics in a reasonable acquisition time?" Another question is "Why do I really need a map?" Mapping is pretty much a cosmetic operation anyway--good for show and tell or for impressing others, but not the way to detect elements at small concentrations. I'd use point or area analyses to get the necessary counting statistics.
You can get the header file for EL/P by using the program's facility for saving/reading files as text. If you're only converting a few files, use cut and paste.
Is Mac Fac a multivariate stat. analysis program? Where can I get it?
Larry Thomas
++++++++++++++++++++++
} From: Tony Garratt-Reed
I don't know what responses were made to Daraporn off the listserver, but there seems to be some confusion in what has been posted on the server.
Daraporn has seen a small peak at the S energy in his EDX spectra, but could not find S in the EELS spectrum. This could be for one of two reasons: 1) the S is not S at all, but Mo or Pb. In this case, with luck, he might expect to see the K-lines of Mo or the L-lines of Pb at higher energy, provided, of course, that there is enough present (the low-energy lines at the S energy are much more intense than the higher energy ones) Mo could certainly be coming from apertures, as Malcolm Haswell suggests. Pb could, conceivably, be coming from the x-ray shielding in the microscope, but do other CM20's have that problem? There is no obvious reason why Daraporn's should if others don't, unless they have made some modification to the stage area. It would seem unlikely that Mo or Pb were actually in the sample. EELS would be used to confirm the analysis because EELS is not susceptible to "hole count" effects. To use the Mo aperture as an example: the Mo x-ray signal is generated by stray radiation (scattered electrons, x-rays)hitting the large area of the mostly thick aperture material. Hence just a few electrons or x-rays can produce lots of Mo x-rays. In contrast, the few electrons that penetrate the aperture would have lost lots of energy and been scattered through large angles, so would not reach the EELS detector at all. There is no equivalent of the mechanism that generated the spurious x-ray signal.
Unfortunately, there is another possible reason for not seeing the S, even if it is really there. The sample could be too thick. The whole relationship of beam current, sample thickness, statistics and spatial resolution is very complex, both in EDX and EELS analysis. However, one point is probably worth mentioning: Comparisons of the detection sensitivities for EDX and EELS compare the results from the same samples. Typically, in practice samples suitable for EELS are thinner than those often used for EDX analysis. Hence, although theory might say that (at least up to Ca or so) the detection limits for EELS are better than those for EDX, this is only true for thin samples. In thicker samples, the EELSbackround rises and the peak becomes blurred out, resulting in poorer detection, while in EDX you just get more signal and hence better detectability.
The papers (from NIST/NBS) that illustrate the sensitivity of EELS also point out the need for extremely high incident beam currents, available only in a LaB6 microscope. I don't know enough about the CM20 FEG to translate from the settings that Daraporn used to actual beam characteristics. However, it is clear that the optimum beam settings for EELS are very different from those for EDX, and also the spatial resolution is significantly degraded. It could be that his microscope conditions are not close to optimum for EELS.
I don't know what is the approx. S-content ... they just told me "it's low" .. but with my parallel probe analysis with 100sec collection time, meter plate reading of 2.5 sec ... the maximum sulfur peak with EDS was ~1000 counts. Not sure what the meter plate is. It might be what I called the processing time in the EDS system. This is essentially the EDS pulse processor time constant. Short processing-times allow high count rates at reduced deadtime, although with some degradation in energy resolution. Long P-times give the best resolution but increase deadtime at a given counting rate. For mapping, you sacrifice energy resolution to allow the highest possible counting rate: set the signal processor for the shortest processing time. But when I switch to do the analysis in STEM mode, because the probe is small, the sulfur peak has to be collected with a longer time. Whatever mode you choose for analysis, you can optimize the counting rate by increasing the probe size and using a large condenser aperture to allow a large beam convergence angle. Nearly all TEMs allow you to change these parameters: you're not limited to the 'standard' microscope settings. You're also not limited to STEM mode. Did Tony Garrett-Read mention that you were using a CM20 FEG? I don't know which EDS system you're using, but most can give at least 30,000 counts./sec with less than 50% deadtime. Sure, you sacrifice spatial resolution, but you'll still get plenty for most jobs, including yours, even with a conventional TEM. To check the sulfur detection, I'd highly recommend spreading the probe to avoid damaging the sample. If you decide to proceed with mapping, first run a little side experiment comparing analyses with a focused probe and a spread one to see if the sulfur is stable. While with EELS, I only need 5 sec or something like that. A 5 second EELS analysis sounds as if you're not counting long enough to get the statistics for a reasonable detection limit.
I need the mapping mainly for the cosmetic purpose and because it's give us spatially resolved information, something like that. That's the term they used.
You mentioned about "point spread in the detection system" in DigiPEELS, what is that? No one mentioned about that. Is that in the newer version? DigiPEELS is Gatan's newest model PEELS detection hardware. If you don't have it, don't worry about it. Newer versions of the EL/P software have a spectrum sharpening function that corrects for the interactions between sampling elements (i.e., pixels) in the digital detectors Gatan uses. If you have this function, check it out in the EL/P software manual or talk to Gatan.
Could you explain briefly how I could optimize EDS acquisition? What are the things to look for? I think I tried to have the shortest data acquisition time and high beam current already. Covered above. A subtle point, perhaps, but the important parameter is probe current rather than total beam current in the microscope. Do you think 2.5 sec meter plate reading is still low? I'm really not too familiar with this.
_____________
Mo peak ... to Dr. Garber and Dr. Malcolm
I don't know about Mo aperture, but I'll check ... but I think the microscope I'm using has Be-window for EDS, is that the movable Mo-aperture you are talking about. Or is it the aperture of the microscope? (Mo) x-rays from the condenser apertures in the microscope can produce spurious signals in the EDS detector. These have no connection with EELS.
One way around the problem of molybdenum when trying to detect sulphur is to use the L2,3 edge of sulphur.
this edge is at 165 eV which is far enough away from the plasmon, so no problems there. other major in that region are
Ho N 45 161 eV Y M 45 157 Dy N 45 154 Tb N 45 147 Gd N 45 140
I hope that these are not in your sample but if they are not strong edges. To get a good signal in this energy range I would have convergence angle of around 11 mrads a COLLECTOR angle of around 3.4 mrads this is the important one
Acquisition set at 0.3 eV per channel 10s integrate time 3 times per acquire attenuator off
The probe should be 1 nm in STEM mode your VSM should have an offset of 100 eV
Sorry, but that Electrohome monitor is not a standard RGB monitor. Ours failed and I remember Kevex telling me that the scan rates were pushed a little harder to get the resolution they used on the Deltas. I tried several RGB monitors including an old Sony TV monitor, a DEC computer monitor, and new Hitachi and NEC computer monitors with RGB inputs. None of them could quite sync up.
We replaced ours with one from another Delta system being retired on campus. I had also asked the list and gotten responses from several others willing to help me out. But now, our Delta is sidelined and I might be persuaded to finally part with the whole thing. Let me know how the search goes.
Waren
At 08:45 AM 11/24/98 -0500, Kalman wrote: } } On Mon, 23 Nov 1998, Craig Franklin wrote: } } I am looking for a used color monitor for a Kevex Delta 1 EDX System. It is } } a model 38-DO5IMA-UU free standing monitor with 5 RCA plugs exiting the } } rear. } } If those plugs are labelled R,G,B,H,V, it is probably a } standard RGB monitor and there are many replacements from } many manufacturers. If it is an older machine, the } bandwidth is probably not great making replacement } relatively easy. Surely, someone in the lab/department has } a computer monitor with RGB inputs you could try out. } } Kal }
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
For what you are going to pay for a darkroom setup (enlarger, supplies, chemicals, paper, etc.) you could afford to go with a digital system. I bought a Polaroid SprintScan 45 for around $8100 (or much less than a good enlarger) This will do full TEM negs at 2000 dpi and 35mm at 4000 dpi. For the full TEM negative, this gives an enlargement factor of about 6.7X if you print to a 300 dpi printer. Although you should really consider getting a sublimation dye printer, you can get marvelous results from $300-$400 inkjet printer. I use an HP 892C at work and a HP 722c at home. Both of these do fantastic photo quality output when used with the Photo Deluxe Paper. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: "740206-at-ucl.itri.org.tw"-at-Sparc5.Microscopy.Com To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Fellow microscopists, I am setting up a darkroom for my electron microscope laboratory recently and looking up a suitable enlarger. I would appreciate your suggestions and comments on what brand/ specifications I need for my TEM, SEM and optical microscopes. Thanks Ren-Jye
I do not belong to this list, however it was suggested that I post my question here. Any help would be greatly appreciated. My e-mail address is jane-at-cc.usu.edu
My question pertains to transmission electron microscopy (TEM) on mammilian embryos. I have done a literature search and will be getting copies of articles soon. However, I was wondering if anyone out in Histoland has processed mammilian embryos for TEM? From the materials and methods of the articles I have seen, they are fixed in standard EM fixative, rinsed, fixed with OsO4, rinsed, dehydrated, and embedded in Epon - all similiar to what I am used to.
Questions: Do you use standard BEEM capsules?
I'll have a dissecting microscope to use when I am processing the samples, however, how do you see them to put them in the block?
Some of the articles I have read suggest embedding the embryos is agar during processing - has anyone done this?
Thank you in advance,
Jane
M. Jane Chambers, MS, LAT Research Technician USDA/ARS Poisonous Plant Research Lab 1150 East 1400 North Logan, UT 84341 Phone: (435) 752-2941 FAX: (435) 753-5681 jane-at-cc.usu.edu
----snip---------- } I wouldn't count on them for long term storage of critical information. we } went to a relatively cheap digital image capture system (Snappy Image } capture card on the NTSC outlet of our AMRAY 1830) and are well pleased with } the results. We typically take both thermal prints and a digital image on } all images. } } John Giles } Principal Materials Engineer } Honeywell Space Systems
"relatively cheap"???? How about DIRT CHEAP!
Seriously though, it brings up a question which I don't think I've heard addressed here even though image capture comes up frequently. I am currently using a Snappy for capturing (on a dinosaur of a computer, I might add!) from an optical scope and am planning on upgrading to a real capture card.
Has anybody compared the consumer capture cards (e.g. All-In-Wonder, for a couple hundred dollars) with the higher cost cards (e.g. Flashpoint, costing a thousand or more)?
I'd be interested in hearing thoughts from the list.
Jim Passmore Analytical Chemist Cryovac Divsion Sealed Air Corporation james.passmor-at-sealedair.com
Rather than purchase a monitor to do this, have you considered using a frame grabber to capture and display the images? You could use a standard PC monitor for viewing the images. This way, you would not only be able to visualize your images, but you'd have the added capability of capturing and storing the images. We have a couple of boards that might do the job, cost is between $1300 - $2500 for non-standard signals. If you can send me specifications for the video signal, I can tell you what you'd need. The boards are auto synching, so all you'd have to do is attach your video cables, and tell the board to determine the signal. This is a very simple procedure which does not require you to program in a bunch of timing values. Feel free to email me if you are interested.
Disclaimer: Obviously, I have a vested interest here. I can solve your problem, but in the process I am looking to sell you this equipment.
Thanks, Jim Haley
****************************** Jim Haley Applications Engineer I-CUBE 2411 Crofton Lane, Suite 14A Crofton, MD 21114 voice: (301) 858-0505 fax: (301) 858-0615 web site: http://www.i-cubeinc.com e-mail: haley-at-i-cubeinc.com ******************************
Craig Franklin wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am looking for a used color monitor for a Kevex Delta 1 EDX System. It is } a model 38-DO5IMA-UU free standing monitor with 5 RCA plugs exiting the } rear. } } If anyone can help, please contact me at 303-689-2224 or e-mail } franklin-at-idcomm.com... } } Thank you, } Craig Franklin } franklin-at-idcomm.com
We have had good luck with our Snappy card. Ours is relatively new (I think the latest version). This is the third or fourth capture board we have had in our system over the last 7-8 years.
Our initial board was a US Video VGA capture board. It was ~$700 and seemed to be oriented towards the TV industry as it had stuff for blue screens and alphanumerics. Quality was okay, not great, but it seemed to be somewhat outdated when we bought it. Chalk that one up to inexperience.
We decided the next board would be more suited towards our technical application. Our budget for this was $1-2K. We chose the Targa 64 card. The resolution was much better than the US Video, but it wasn't the most user friendly software and we had all grades of problems when we went to a Windows operating system (from DOS).
The next card was a Snappy. the resolution was as good as the Targa and the software was much more user friendly. We had this one for a while and then purchased a later version since it was so cheap (~$100).
In our application, the high end boards don't do a lot for us, since we are getting an NTSC video signal out of our AMRAY 1830. Our other option would be a retrofit with a system to capture the digital signal, but we haven't reached that step yet in either need or budget.
John Giles Principal Engineer Honeywell Space Systems
} "relatively cheap"???? How about DIRT CHEAP!
} Seriously though, it brings up a question which I don't } think I've heard addressed here even though image } capture comes up frequently. I am currently using a } Snappy for capturing (on a dinosaur of a computer, I } might add!) from an optical scope and am planning on } upgrading to a real capture card.
} Has anybody compared the consumer capture cards (e.g. } All-In-Wonder, for a couple hundred dollars) with the } higher cost cards (e.g. Flashpoint,costing a thousand or } more)?
} I'd be interested in hearing thoughts from the list.
} Jim Passmore } Analytical Chemist } Cryovac Divsion } Sealed Air Corporation } james.passmor-at-sealedair.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for a used color monitor for a Kevex Delta 1 EDX System. It is a model 38-DO5IMA-UU free standing monitor with 5 RCA plugs exiting the rear.
If anyone can help, please contact me at 303-689-2224 or e-mail franklin-at-idcomm.com...
by rwja.UMDNJ.EDU (8.8.6 (PHNE_14041)/8.8.6) with ESMTP id SAA16673; Tue, 24 Nov 1998 18:04:52 -0500 (EST) Message-ID: {365B64B5.80DF1B0C-at-umdnj.edu}
740206-at-ucl.itri.org.tw-at-sparc5.microscopy.com wrote: } } Fellow microscopists, } I am setting up a darkroom for my electron microscope laboratory recently and looking up a suitable enlarger. I would appreciate your suggestions and comments on what brand/ specifications I need for my TEM, SEM and optical microscopes. } Thanks } Ren-Jye
My choice would be a Beselar 45MXT enlarger, about $1000. Will do everything from 35 mm to 4 by 5 film. Be sure to get a high quality enlarging lens, such as an El Nikor or Schneider which will cost about $250-$300. A 135 mm lens is the correct focal length for 3.25 by 4 EM film. You can add other lenses to the system as you need them. With reasonable care such equipment will last forever.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Evex Service, a division of Evex Analytical, offers repair exchange = parts and detector upgrades for many older and some newer style x-ray = analyzers. Evex Service, maintains a full line of Kevex, Tracor, Noran, = and PGT Analyzers for its Service Contract Customers. If you need parts = or technical support, feel free to contact any of our experienced = service engineers, the average has 10+ years in x-ray microanalysis.
Peter Tarquinio Evex Analytical 857 State Road Princeton, NJ 08540
-----Original Message----- } From: Gary M. Easton {gary.easton-at-scannerscorp.com} To: MSA Listserver {microscopy-at-sparc5.microscopy.com}
Dear Colleagues:
Once again I have found myself with a few extra hotel rooms for a conference. I have 2 rooms available for the Marriott in Boston for the Materials Research Society Meeting. The rooms are available for arrival Saturday November 28 for 6 nights at the conference rate which I believe = is $128.
I made this same offer for the MSA meeting and realized that many people for one reason or another need last minute hotel rooms. If that is the case for MRS meeting, please let me know immediately. If I don't have an= y takers, I'll be canceling the rooms tomorrow night.
I am making some 200 mesh coated nickel grids for some immuno work. Normally we use Pioloform from Agar Scientific to make our support films, but I have never used coated grids for immuno work before. Does anyone have experience using Pioloform-coated grids for immuno staining?
If so will they work OK or will I have to resort to Formvar ?
{html} {!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=text/html;charset=iso-8859-1 http-equiv=Content-Type} {META content='"MSHTML 4.72.3110.7"' name=GENERATOR} {/HEAD} {BODY bgColor=#ffffff} {DIV} {FONT color=#000000 size=2} Hi, {/FONT} {/DIV} {DIV} {FONT color=#000000 size=2} I'm in need of a EDS HV bias module, NIM style, with an adjustable output. I can't use the one made by Tracor as it has to be paired with their pulse processor. Maybe one out of an old KEVEX 5100 system or similar. Thanks {/FONT} {/DIV} {DIV} {FONT color=#000000 size=2} {/FONT} {/DIV} {DIV} {FONT color=#000000 size=2} {/FONT} {/DIV} {DIV} {FONT color=#000000 size=2} Gary M. Easton, Pres. {/FONT} {/DIV} {DIV} {FONT color=#000000 size=2} SCANNERS CORPORATION {/FONT} {/DIV} {DIV} {FONT color=#000000 size=2} Third Party SEM Service/Maintenance {/FONT} {/DIV} {/BODY} {/HTML}
I would like to thank all those that responded to my question. I now feel that I know more than I did before I asked (heh, imagine that...). As time permits, I may correspond with those offering their expertise or services. To the rest, thank you very much. Have a wonderful Thanksgiving Holiday!!
Best Regards,
Tim tim_wallace-at-doh.state.fl.us
Go FSU Seminoles! (oops, sorry about that.) ________________________________________________ Tim Wallace, Environmental Specialist II Florida Department of Health Volusia County Health Department, Environmental Health Division Indoor Air Assistance / Lead Monitoring Programs 501 S. Clyde Morris Blvd., Daytona Beach, Florida 32114 USA phone: 904.947.3484 / fax: 904.947.3485 http://www.state.fl.us/cf_web/D12/cphu/ehprgms/iaq.html +++++++++++++++++++++++++++++++++++++
P.S. Just in case that anyone is interested, there is a listserve on the topic of Indoor Air Quality called iaq-at-onelist.com. A subscription can be had at {http://www.onelist.com}, go to "find a list", go to "environmentalism" and dbl. click, scroll down to the IAQ list description. There you can subscribe if interested. I have no financial interest in the IAQ listserve.
Hi all Re the thread on HEPES as buffer vehicle for glut fixation:
A long time ago we did some work on the buffering capacity of some common (and a few not-so-common) buffers. HEPES was one of these. This buffer has a Pka right in the desired area (approx 7.5) and has reasonable buffering capacity. Cost is the primary disadvantage.
The reference is: Coetzee & van der Merwe (1987): Some characteristics of the buffer vehicle in glutaraldehyde-based fixatives. Journal of Microscopy, 146, 143-155.
Jan Coetzee
} I've heard that this buffer does not hold it's buffering capacity very } well. though I've no personal experience with it. } } Perhaps someone out there has worked with it.
Prof Jan Coetzee Head: Lab for Microscopy and Microanalysis Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-362-5150 Pretoria 0002, South Africa http://www.up.ac.za/science/electron/emunit1.htm
I could not access email for a while due to network problems in my institute. For the question I asked about carbon rod sharpener, many people sent me good pieces of information, suggestion, or even a free rod sharpener. They were all very helpful to me. Thank you all who helped me and thank ListServer.
Especially many thanks to: Dr Laurence Tetley at University of Glasgow Olli Taikina-aho at University of Oulu, FINLAND Malcolm Haswell at University of Sunderland Dr. Ming H. Chen at University Of Alberta, Canada John Heckman, MSU Center for Electron Optics James Young David Henriks, South Bay Technology, USA Bill Tivol Markus F. Meyenhofer, Microscopy Labs, USA Paul Nolan
Jondo Yun Department of Inorganic Materials Engineering Kyungnam University 449 Weolyeong-dong Masan, 631-701 Korea 82-551-249-2697 (office) 82-551-248-5033 (fax) 82-551-249-2692 (department office) 82-551-249-2719 (laboratory) 82-551-249-2564 (EM lab) email: jdyun-at-hanma.kyungnam.ac.kr
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In response to Ren-Jye who had questions about conventional darkrooms, I have a suggestion that might make your work easier. Micrographs of metals taken on a TEM tend to have high contrast when developed in D-19, making printing difficult. I switched to Acufine Co.'s Diafine developer. It requires soaking the film 3 min. in each of two solutions, then fixing and rinsing normally. It provides CHEMICAL DODGING in effect, reducing contrast to a very printable level. It has good shelf life as mixed and is more expensive-but the time and print paper saved make up for that. I normally can print negs. on No. 2 contrast paper. Bernie Kestel {Kestel-at-anl.gov}
(1.37.109.15/16.2) id AA140123896; Wed, 25 Nov 1998 11:11:36 -0600 Posted-Date: Wed, 25 Nov 1998 11:11:36 -0600 Received-Date: Wed, 25 Nov 1998 11:11:36 -0600
A few weeks ago I posted a question about etching glycol methacrylate polymer (specifically JB4, Polysciences) for immunohistochemistry in small tissue aggregates. I'd like to thank Hank Adams for suggesting an etching technique that worked. I was able to see specific staining in my samples using his suggestion.
} Try etching for 3-5 minutes in sodium ethoxide soln 1:1 with toluene, } then go to 100% EtOH down to water, buffer, etc. } } Ethoxide Solution: } 75gm NaOH in 946 ml absolute ethanol; let stand 10-14 days or until } pellets dissolve with occasional stirring. Solution should turn brown } when ready.
} Hank Adams } Cell Biology } Integrated Microscopy Core } Baylor College of Medicine } One Baylor Plaza } Houston, Tx 77030
Susan A. Fugett
Department of Chemical Engineering and Materials Science Phone: 612-625-8803 University of Minnesota 612-625-0808 421 Washington Ave SE Fax: 612-626-7246 Minneapolis, MN 55455 Email: fugett-at-cems.umn.edu
Robert Santoianni {Robert_Santoianni-at-emory.org} wrote:
} To Harris Reavin, } The evolution of clearing solvents is interesting. Toluene (actually } chloroform) works best, but is a no-no because of carcinogenicity and } environmental (disposal) problems. Xylene has been the solvent of } choice for both clearing and dewaxing, but again, health effects and } disposal are major concerns. Limonene products came on the scene as } the "magic" xylene substitute, boasting the fact(?) that they could be } dumped down the drain. Well, in my experience they don't perform well, } our local sewer authorities don't want them in the system and you can } smell them out in the parking lot! Aliphatic hydrocarbons, such as certain } products from Richard-Allen Scientific 800-522-7270 and ANATECH } 800-ANATECH have a proven track record as xylene substitutes. Check } with these companies, you might persuade them to send you some } samples...and they both are very knowledgeable and helpful and offer all } kinds of expert advice. In my opinion, you'll get good clearing with } aliphatic hydrocarbons, but you will have to do your whole processing } procedure under a fume hood. Remember gloves, goggles and } apron...be safe! } Good luck, } Bob Santoianni } Emory University Hospital } Atlanta, GA } robert_santoianni-at-emory.org
I have recently begun using limonene as a clearing agent, and have received decent results. I am looking at cadaveric tissue that has been in phenol for a while (years), but I've been able to get decent preservation using limonene as a clearing agent. I'm also working around students who are not professionals used to working with more dangerous sovents, so I feel a little safer giving them limonene. As for odor, like many people with a biochem background, I have a weak sense of smell, so I don't really notice it.
I am aware my needs are very different than a path lab making surgical decisions based on their results, and I'm also not doing this work full time. If I was doing it day in and day out, poorer preservation and overwhelming odor might be more of an issue.
Also, I'm interested in the body of research suggesting limonene has antitumour effects. While I do not take a nip out of the bottle or intentionally inhale fumes, I sometimes wonder if it might help neutralize the "sins of my youth"(exposure to aldehydes, Osmium and what not). What are others experiences with the magic orange juice? Given it's low flash point, is there a safe way to dispose of it by burning it? I'm not suggesting this, just curious if it's been tried or thought out.
Also, does anyone know if its clearing properties deteriorates if BHA or BHT isn't added? Or what the optimal concentration of these additives is? I'm also looking into cheaper sources. I can get 55 gallons of a higher grade of limonene from a chemical company for less than half of what one of the histology companies is charging for five gallons, but I don't really feel like storing 55 gallons of a flammable in a small lab.
Charlie Ginsburg NCC Research Dept. Lombard IL
_________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
Dear List I have the opportunity to purchase an inverted fluorescent microscope,to use for viewing cells infected with a virus expressing green fluorescent protein, and to take photographs or capture an image that will let me count plaques, measure intensity of the signal etc. As this has to live in the virus room, it may not be practical to have a multipurpose computer set up alongside, so I was thinking about a digital camera with discs so users could take information away to analyse elsewhere. I am inundated with options but I wonder if anyone has such a setup that they are happy with and could recommend. My budget is =A325K absolutely max. Please respond off the list and I will summarise at a later date! Many thanks Vivien Mautner
Vivien Mautner CRC Institute for Cancer Studies The University of Birmingham Edgbaston BIRMINGHAM B15 2TA
while we can't help you with the diffractometer and it's components, we can probably help you with your CCD camera, image acquisition and processing needs. We have a camera that allows to acquire 10-bit images with extremely short or very long exposure times to cover the wide dynamic range of diffraction patterns.
If you are interested in getting more info, please contact me directly.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS fax: (303) 234-9271 email: info-at-soft-imaging.com
================== Disclaimer: Soft imaging System Corp produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. ==================
--------- Forwarded Message ---------
DATE: Mon, 23 Nov 1998 08:02:32 } From: "Dennis C. Winkler" {Dennis_Winkler-at-nih.gov} To: Microscopy-at-Sparc5.Microscopy.Com
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Hello all,
We are planning to rebuild the optical diffractometer we use to assess the quality of our TEM micrographs. Anyone have a favorite design or modification for an optical diffractometer (and perhaps an associated reference)? We are looking to build a vertical set up similar to the one in:
We'd also like to add a CCD camera with a high quality thermal printer.
Any recommendations?
Thanks,
- Dennis
------------------------------------------------------------------------ ------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
--------- End Forwarded Message ---------
-----== Sent via Deja News, The Discussion Network ==----- http://www.dejanews.com/ Easy access to 50,000+ discussion forums
I've replaced our Targa framegrabber with the Snappy ($3000 vs. $100) and am much happier with the Snappy. Simple to setup and simple to use. Trying to figure out the Targa manual set (yes, set!) is no picnic. The Snappy manual alone is worth the price, simply for entertainment value. Even their licensing agreement is hilarious. For standard NTSC resolution cameras, I wouldn't bother with anything else.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ----snip---------- } } I wouldn't count on them for long term storage of critical information. we } } went to a relatively cheap digital image capture system (Snappy Image } } capture card on the NTSC outlet of our AMRAY 1830) and are well pleased with } } the results. We typically take both thermal prints and a digital image on } } all images. } } } } John Giles } } Principal Materials Engineer } } Honeywell Space Systems } } "relatively cheap"???? How about DIRT CHEAP! } } Seriously though, it brings up a question which I don't think I've heard } addressed here even though image capture comes up frequently. I am } currently using a Snappy for capturing (on a dinosaur of a computer, I } might add!) from an optical scope and am planning on upgrading to } a real capture card. } } Has anybody compared the consumer capture cards (e.g. All-In-Wonder, } for a couple hundred dollars) with the higher cost cards (e.g. Flashpoint, } costing a thousand or more)? } } I'd be interested in hearing thoughts from the list. } } Jim Passmore } Analytical Chemist } Cryovac Divsion } Sealed Air Corporation } james.passmor-at-sealedair.com
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
A year or so ago the address for Taab Labs Equip. Ltd. was 3 Minerva House, Calleva Industrial Park, Aldermaston, Berkshire RG7 4QW, UK, Tel: 0734-817775, Fax: 0734-817881
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
I could not access email for a while due to network problems in my institute. For the question I asked about carbon rod sharpener, many people sent me good pieces of information, suggestion, or even a free rod sharpener. They were all very helpful to me. Thank you all who helped me and thank ListServer.
Especially many thanks to: Dr Laurence Tetley at University of Glasgow Olli Taikina-aho at University of Oulu, FINLAND Malcolm Haswell at University of Sunderland Dr. Ming H. Chen at University Of Alberta, Canada John Heckman, MSU Center for Electron Optics James Young David Henriks, South Bay Technology, USA Bill Tivol Markus F. Meyenhofer, Microscopy Labs, USA Paul Nolan
Jondo Yun Department of Inorganic Materials Engineering Kyungnam University 449 Weolyeong-dong Masan, 631-701 Korea 82-551-249-2697 (office) 82-551-248-5033 (fax) 82-551-249-2692 (department office) 82-551-249-2719 (laboratory) 82-551-249-2564 (EM lab) email: jdyun-at-hanma.kyungnam.ac.kr
Our group currently has a JEOL FX 2000 TEM available for sale to anyone interested. The system includes a cold and hot stage, PC driven X-ray system,and STEM attachment. Please respond if interested.
M.W. Rigler, Ph.D. MAS, Inc. Suwanee GA 770-866-3218
I am currently in the process of making cross-section samples of a machined Ni-Cr alloy. The samples are first cleaned in NaOH (& Al. clip) and rinsed in warmed distilled H2O. They are then ultrasonically cleaned in distilled acetone, and also suspended in an acetone reflux vapor degreasing for 20 minutes.
I immerse the samples in the Nickel "Electroless" plating solution for ~20 minutes at 90-100 C. Fine bubbles form indicating that plating is proceeding but when I remove the sample, a discolouration appears on the surface and the film doesn't appear to adhere very well.
Any suggestions?
I eventually want to Nickel electroplate to about 2-3 mm. then prepare the cross-sections.
thanks in advance
Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
I don't know if there is anything commercially available for this for light microscopes, but you might want to check with Vince Carlino at VCR, Inc. (VCRvince-at-aol.com) Their D500i dimpler has a non-contacting sensor that has 1 um resolution. This system does have a feedback loop. He might be able to help you or direct you to someone that can. -Scott Walck
---------- } From: Bruce Brinson To: MSA Listserver -----------------------------------------------------------------------.
Hello photon fans, I need an close loop, contactless system that can check & correct the working distance between objective & glass plate to within 1u. I would appreciate any feed back this group can provide. Please contact me directly if your company has markets this type of product.
Dear sir, It will be grateful if you let me know if you have an available second hand TEM. My Fax is 009821 8895980 or 009821 2548220 Tank you very much. Hmid. habibi hrhb4-at-nrcgeb.ac.ir
The Materials Research Science and Engineering Center at the University of Houston has an opening for a research faculty member or postdoctoral fellow to manage and conduct research in their newly completed JEOL 2010F FEG-TEM facility. The facility became operational in July 1998 and consists of a JEOL 2010F FEG-TEM (high-resolution pole piece, 1.9=C5 point-to-point resolution verified), with Gatan 794 Multi-scan CCD Camera, Gatan 666 PEELS, and X-ray EDS system to be purchased in early '99. An additional JEOL 2000FX analytical TEM is available as part of shared facilities with the Texas Center for Superconductivity, together with complete sample preparation facilities.
MRSEC-UH is an NSF-funded center dedicated to research on advanced oxides for solid-state ionics applications and a background in TEM-based oxide structural chracterization is desirable but not essential. The successful candidate will conduct cooperative research related to the various active projects in the Center, and will manage the day-to-day use of the facility. Development of the facility's full potential in the area of HRTEM imaging and nano-probe microanalysis will be expected.
Interested individuals should respond (off-line please!) to:
Professor Allan J. Jacobson, Director MRSEC-UH Department of Chemistry University of Houston, Houston, TX 77204 ajjacob-at-uh.edu
Additional information regarding MRSEC-UH can be found at: www.uh.edu/mrsec. The University of Houston is an Equal Opportunity/Affirmative Action Employer. Minorities, women, veterans and persons with disabilities are encouraged to apply.
We are faced with replacing an ancient Haskris on a diff-pumped Balzer 301. Wondering whether Haskris is still the way to go, and whether there are reliable alternatives. We are working with a water-cooled compressor...Thanks...Tom Reese
Dear driver experts, Our laboratory have a old monitor Sony, model GDM 1952 Trinitron. What kind of graphic cart and driver we can use for our windows applications Many thanks OPEA 114, rue de la Jarry 94300-VINCENNES (FRANCE) e-mail: opea.hoan-at-wanadoo.fr
I'm studying arthropod (mostly small crustacean) appendages. Some of the mounts are relatively thick, with fine structures well below the coverslip. I've read Barron's fine book and noted his comments on thick mounts causing spherical aberration. I have considered modifying my scope to have adjustable tubelength. But it occurred to me that some correction could be obtained by using a high index mounting medium, i.e., higher than immersion oil / coverslip... and higher than chitin to get contrast. Cargille's "Meltmount" sounds good but I prefer a water soluble medium. Questions follow. --- Am I correct in my belief that a high index medium will nullify some spherical aberration in these thick mounts? --- One type of "high index" medium uses Chloral Hydrate, which is hard to obtain and I doubt the index is very high. I've heard recommendations of heavy halogen salts... any recipes will be appreciated. --- I've been using mostly gum - glycerin - water. Another problem there is shrinkage with distortion of the coverslip, which can't be good for resolution. Any hints to minimize that will be appreciated. --- The above medium is a colloid(?); am I losing contrast due to light scatter? Would a true solution be *significantly* better with brightfield? Darkfield? --- Does anyone know the refractive index of chitin? (Looks barely above imm. oil). How much higher should the medium index be to get good contrast AND resolution... without too much Becke line / white line?
I hope no one minds these low - tech questions on a high - tech newsgroup. I've done a fair amount of web-search without finding the information I'm looking for.
I shall be on leave during 3-20 Dec. I will switch on the auto-answer function (every coming e-mail will get auto-answer from my computer) and I do not want this auto-answer affect every member of this mail list. So I want to unsubscribe this mail list. Please take my name from the mail list. Thank you very much.
-----Original Message----- From: malcolm.haswell-at-sunderland.ac.uk [SMTP:malcolm.haswell-at-sunderland.ac.uk] Sent: Friday, November 13, 1998 1:21 AM To: Subject: Re: To CPD or to HMDS, that is the ques
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Could something be sent to this list too please - it would certainly
interest me.
thanks
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: ROBIN CROSS To: Paula Sicurello Cc: microscopy Subject: Re: To CPD or to HMDS, that is the ques Date: 12 November 1998 15:51
Hello Paula
} Or I was considering HMDS, has anybody out there used HMDS for } things as delicate as cells?
Shirley Pinchuck, in this lab, has done a fair amount of work using HMDS, including comparisons of HMDS with other methods such as CPD, cryo-SEM, etc. I will ask her to fax you her protocols as well as an abstract of a conference presentation on some of the comparative work.
I hope this helps.
Regards
Robin
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
} -----Original Message----- } From: ZHANG Tiejun } Sent: Monday, November 30, 1998 6:11 PM } To: DAI Jiyan; DAI Jiyan } Subject: EM survey } } Dear all: } I post this message for a TEM service engineering. } } Please kindly let me know which EM Lab has 3 sets of TEM made by JEOL. } Thank you in advance!
Dear driver experts, Our laboratory have a old monitor Sony, model GDM-1952 Trinitron with input RGB, H.synchro. and V.synchro. What kind of graphic card and driver we can use for our windows applications Many thanks Hoan OPEA 114, rue de la Jarry 94300-VINCENNES (FRANCE) e-mail: opea.hoan-at-wanadoo.fr
I suppose that you could use the monitor on most any card with an RGB output. However, I don't know how many windows applications there would be that would make use of it. It might be that frame grabber/display cards could make use of it in connection with image analysis programs. Matrox and other companies could probably answer those questions for you and let you know what products they have that would drive that monitor. Their cards would likely come with the necessary drivers.
I am not familiar with the size of that monitor or its resolution specifications. If you are thinking of using it as your main monitor, you may well be able to find or make an adapter cable to go from a standard VGA connector to the BNC connectors you probably need for RGB. However, there would likely be issues of the monitor being able to run at the scan frequencies that VGA cards support. The resolution may also be unsatisfactory. Very old VGA monitors specified a dot pitch of 0.39 mm. Then came monitors with 0.28 mm pitch and now 0.22 mm. If your Sony does not have such a fine pitch, you would probably find the graphics blurred.
That's about as much as I can tell you.
At 01:34 PM 11/30/98 +0100, you wrote: } } Dear driver experts, } Our laboratory have a old monitor Sony, model GDM-1952 Trinitron with } input RGB, H.synchro. and V.synchro. What kind of graphic card and } driver we can use for our windows applications } Many thanks } Hoan } OPEA } 114, rue de la Jarry } 94300-VINCENNES (FRANCE) } e-mail: opea.hoan-at-wanadoo.fr
It is true that, for large angles of incidence, the spherical aberration of a plate is proportional to t/N (thickness over refractive index). So higher index might just give you a better image, if the other aberrations don't creep up on you, particularly longitudinal chromatic. But wouldn't you want a higher index cover slip as well?
best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
Hello all;
I'm studying arthropod (mostly small crustacean) appendages. Some of the mounts are relatively thick, with fine structures well below the coverslip. I've read Barron's fine book and noted his comments on thick mounts causing spherical aberration. I have considered modifying my scope to have adjustable tubelength. But it occurred to me that some correction could be obtained by using a high index mounting medium, i.e., higher than immersion oil / coverslip... and higher than chitin to get contrast. Cargille's "Meltmount" sounds good but I prefer a water soluble medium. Questions follow. --- Am I correct in my belief that a high index medium will nullify some spherical aberration in these thick mounts? --- One type of "high index" medium uses Chloral Hydrate, which is hard to obtain and I doubt the index is very high. I've heard recommendations of heavy halogen salts... any recipes will be appreciated. --- I've been using mostly gum - glycerin - water. Another problem there is shrinkage with distortion of the coverslip, which can't be good for resolution. Any hints to minimize that will be appreciated. --- The above medium is a colloid(?); am I losing contrast due to light scatter? Would a true solution be *significantly* better with brightfield? Darkfield? --- Does anyone know the refractive index of chitin? (Looks barely above imm. oil). How much higher should the medium index be to get good contrast AND resolution... without too much Becke line / white line?
I hope no one minds these low - tech questions on a high - tech newsgroup. I've done a fair amount of web-search without finding the information I'm looking for.
I was wondering if it would be easier to find an objective with a correction collar on it? We have a 40x with a collar that we use for all our samples that are outside the standard thickness. It helps a lot. But I don't know if it would give enough resolution for your situation.
Bob Derm Imaging Center U of W
On Sun, 29 Nov 1998 Sjoplinh-at-aol.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all; } } I'm studying arthropod (mostly small crustacean) appendages. Some of the } mounts are relatively thick, with fine structures well below the coverslip. } I've read Barron's fine book and noted his comments on thick mounts causing } spherical aberration. } I have considered modifying my scope to have adjustable tubelength. But } it occurred to me that some correction could be obtained by using a high index } mounting medium, i.e., higher than immersion oil / coverslip... and higher } than chitin to get contrast. Cargille's "Meltmount" sounds good but I prefer } a water soluble medium. Questions follow. } --- Am I correct in my belief that a high index medium will nullify some } spherical aberration in these thick mounts? } --- One type of "high index" medium uses Chloral Hydrate, which is hard to } obtain and I doubt the index is very high. I've heard recommendations of } heavy halogen salts... any recipes will be appreciated. } --- I've been using mostly gum - glycerin - water. Another problem there is } shrinkage with distortion of the coverslip, which can't be good for } resolution. Any hints to minimize that will be appreciated. } --- The above medium is a colloid(?); am I losing contrast due to light } scatter? Would a true solution be *significantly* better with brightfield? } Darkfield? } --- Does anyone know the refractive index of chitin? (Looks barely above } imm. oil). How much higher should the medium index be to get good contrast AND } resolution... without too much Becke line / white line? } } I hope no one minds these low - tech questions on a high - tech newsgroup. } I've done a fair amount of web-search without finding the information I'm } looking for. } } Regards, } } Scott Harden sjoplinh-at-aol.com } } }
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11/30/98 9:00 AM
A friend of mine working on a similar nickel plating problem years ago did not have good luck with electroless nickel plating-it did not adhere well. He made test runs with electroplating only on scrap material first. The general procedure was to have the specimen immersed in the plating bath, start with a low current and voltage in REVERSED polarity mode first for a few seconds, then change to the plating mode polarity and plate for several hours at a slow rate for good adhesion and fine grain,(again at a low voltage/current setting). This process is called the "strike" in library books. Then a higher voltage and plating rate may be used to build up a useful nickel plating thickness. This sequence resulted in good adhesion to the specimen. The Ph of the solution had to be kept near some optimum value. A bag of nylon polishing cloth was tied around the sacrificial electrode to keep residues from moving to the specimen being plated. Reading up on the process shoul! d provide the needed details.
Bernie Kestel {kestel-at-anl.com}
--------------------------------------
Greetings:
I am currently in the process of making cross-section samples of a machined Ni-Cr alloy. The samples are first cleaned in NaOH (& Al. clip) and rinsed in warmed distilled H2O. They are then ultrasonically cleaned in distilled acetone, and also suspended in an acetone reflux vapor degreasing for 20 minutes.
I immerse the samples in the Nickel "Electroless" plating solution for ~20 minutes at 90-100 C. Fine bubbles form indicating that plating is proceeding but when I remove the sample, a discolouration appears on the surface and the film doesn't appear to adhere very well.
Any suggestions?
I eventually want to Nickel electroplate to about 2-3 mm. then prepare the cross-sections.
thanks in advance
Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
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Replying to Barry Shaw's question about using Pioloform-coated grids---Pioloform will do just fine in immuno procedures. As with Formvar, you will see some of the same problems with electrostatic binding of immunogold probes to the film, but they may be prevented by incorporating a surfactant such as Tween-20 (0.05-0.1%) to your dilution and wash buffers or by using a high salt concentration in the wash buffers after the incubation in immunogold. If you are using the Pioloform to support sections instead of cells or cell fractions, the background on the Pioloform film alone may not interfere, and as long as the sections are free of background, you may not need to use a surfactant or otherwise modify the washing procedure.
Vicky
Victoria J. Madden Microscopy Services Laboratory Dept. of Pathology and Laboratory Medicine University of North Carolina-Chapel Hill vmadden-at-med.unc.edu
(SMTPD32-4.06) id AA69A80009A; Mon, 30 Nov 1998 11:40:09 EDT Message-Id: {3.0.3.32.19981130115108.0077ae70-at-mail.map.com} X-Sender: mme-at-mail.map.com X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.3 (32)
Dear Scott,
You have asked a series of interesting questions.
First, the whole contrast problem is derived from the similarity of the refractive indices of chitin and your mounting material. I did some work on this issue many years ago and offer the following solutions:
1. First: an observation:
I agree with you that the refractive index of chitin is about the same as immersion oil (1.515). As I remember, we tried immersion oil as a mounting material and the chitin all but disappeared.
2 Re: refractive index changes
You don't have to have much of a refractive index change to increase the contrast. You can either go up or go down. Cargille has a whole slew of mounting oils from which to choose. You also mentioned that you would like something water soluble. How about just water (ri = 1.33)? You may be able to do a more permanent mount by making sure that the drop of water does not exceed the edge of the coverslip then sealing with something like nail polish.
3. Re:spherical aberration. The microscope's optics expect to see very specific components in the sample prep "sandwich". All the changes which cause spherical aberration (amoung other things) are derived from Snell's law (see any basic physics book). A higher refractive index will acutally accentuate the problem, not solve it. The first thing to do to optimize imaging is to check the barrel of the objectives you are using and make sure that they say "0.17" for coverslip thickness (to order: a number 1-1/2 is the equivalent). Anything thicker will contribute significantly to spherical aberration. Secondly, if you begin to get into the higher ri mounting materials, you may want to try an objective with a "coverslip correction collar". Adjusting the collar may reduce the spherical aberration problem.
3. Colloids are, indeed, serious scatterers. The resulting scatter will create glare and haze which obscures information.
4. I don't understand the "shrinkage with the coverslip" issue. Are you talking about the size of the image or a change in the prep over time?
For more specifics on coverslips, mounting media, and spherical aberration, may we suggest the book "Optimizing Light Microscopy for Biological and Clinical Laboratories"? Details are available at our website: { {http://www.MME-Microscopy.com/education}
If any of you can help this young lady, please respond to her directly, as she does not have access to the listserve.
Best regards, Steven Slap, Energy Beam Sciences
Dear all,
I am resending this message because of a "nondelivery" warning on the first try. Please ignore if you have already received it.
Thanks,
Barbara Foster
Dear Scott,
You have asked a series of interesting questions.
First, the whole contrast problem is derived from the similarity of the refractive indices of chitin and your mounting material. I did some work on this issue many years ago and offer the following solutions:
1. First: an observation:
I agree with you that the refractive index of chitin is about the same as immersion oil (1.515). As I remember, we tried immersion oil as a mounting material and the chitin all but disappeared.
2 Re: refractive index changes
You don't have to have much of a refractive index change to increase the contrast. You can either go up or go down. Cargille has a whole slew of mounting oils from which to choose. You also mentioned that you would like something water soluble. How about just water (ri = 1.33)? You may be able to do a more permanent mount by making sure that the drop of water does not exceed the edge of the coverslip then sealing with something like nail polish.
3. Re:spherical aberration. The microscope's optics expect to see very specific components in the sample prep "sandwich". All the changes which cause spherical aberration (amoung other things) are derived from Snell's law (see any basic physics book). A higher refractive index will acutally accentuate the problem, not solve it. The first thing to do to optimize imaging is to check the barrel of the objectives you are using and make sure that they say "0.17" for coverslip thickness (to order: a number 1-1/2 is the equivalent). Anything thicker will contribute significantly to spherical aberration. Secondly, if you begin to get into the higher ri mounting materials, you may want to try an objective with a "coverslip correction collar". Adjusting the collar may reduce the spherical aberration problem.
3. Colloids are, indeed, serious scatterers. The resulting scatter will create glare and haze which obscures information.
4. I don't understand the "shrinkage with the coverslip" issue. Are you talking about the size of the image or a change in the prep over time?
For more specifics on coverslips, mounting media, and spherical aberration, may we suggest the book "Optimizing Light Microscopy for Biological and Clinical Laboratories"? Details are available at our website: { {http://www.MME-Microscopy.com/education}
Chitin should have an index close to cellulose, which is listed as 1.55 (Merck). Chloral is a controlled substance. I have seen references to use of KI in water, water--glycerol, and a commercial resin but no recipes. 40% KI in water only has n 1.4027. HgI2 can be added if you can live with that (will check reference at home).
There must be polymers with minimum light scattering. I have used surgical jelly, but it contains some junk, which can be removed with a membrane filter, as a result of which the viscosity changes.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
In a message dated 98-11-30 12:08:07 EST, Barbara write:
{ {... 3. Re:spherical aberration. The microscope's optics expect to see very specific components in the sample prep "sandwich". All the changes which cause spherical aberration (amoung other things) are derived from Snell's law (see any basic physics book). A higher refractive index will acutally accentuate the problem, not solve it. { {
(Snell's sounds familiar... maybe like Barron's "sine law".) I got a headache trying to figure which would correct for a thick mount: high or low index. It seems simpler to imagine a perfect image at the eyepiece field stop, and that *projected* back to the object, which is too far below the coverslip. With a low or standard index the marginal rays converge above the object; a high index refracts them more (toward the "normal") so they should converge closer to, if not at, the object. But, I'm not at all certain of that!
} } Secondly, if you begin to get into the higher ri mounting materials, you may want to try an objective with a "coverslip correction collar". Adjusting the collar may reduce the spherical aberration problem. { {
I doubt I'll ever get that type objective, but I may modify my scope to have adjustable tubelength, which apparently does +/- the same thing. Easy on my scope as it's a monocular.
} } 3. Colloids are, indeed, serious scatterers. The resulting scatter will create glare and haze which obscures information. { {
I was afraid of that.
} } 4. I don't understand the "shrinkage with the coverslip" issue. Are you talking about the size of the image or a change in the prep over time? { {
As these mounts dry, the coverslip (std. thickness) is "mounded" (convex) over the object. This is easy to see, e.g., looking at the reflected image of a lamp (naked eye). This wouldn't matter with immersion and a n = 1.51 mount medium. With dry objectives it would seem a disaster --- the coverslip has become an unwelcome lens. I assume "Meltmounts" (and similar) are a "total solids" medium and will not shrink... but I've been hoping to avoid all the associated hassles of non- water miscible media.
} } For more specifics on coverslips, mounting media, and spherical aberration, may we suggest the book "Optimizing Light Microscopy for Biological and Clinical Laboratories"? Details are available at our website: { {http://www.MME-Microscopy.com/education}
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Many people expressed an interest in my recent posting regarding = immunocytochemical methods. For an intense immersion in = immunocytochemical techniques (cryosectioning, resins, LM, EM, stereology) = may I recommend a FEBS-sponsored course to be held in Oslo in June 1999. = Details can be found at: {http://www.hei.org/htm/curs.htm}
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/apw.htm
by imo14.mx.aol.com (IMOv16.10) id 3GVKa26092; Mon, 30 Nov 1998 14:56:33 -0500 (EST) Message-ID: {3be829a0.3662f871-at-aol.com}
I wrote, in response to Barbara Foster's helpful information;
{ { It seems simpler to imagine a perfect image at the eyepiece field stop, and that *projected* back to the object, which is too far below the coverslip. With a low or standard index the marginal rays converge above the object; a high index refracts them more (toward the "normal") so they should converge closer to, if not at, the object. But, I'm not at all certain of that! } }
Well, I'm pretty certain that I was wrong and Barbara is right! Sorry about that and the waste of bandwidth. I think I see what I neglected to consider... embarrassing.
So I'll be using low index media for the thick mounts. That's good and bad news: The bad news is, I probably can't do much better than glycerin (n ~ 1.46) for resolution. The good news is, these things look pretty sharp in glycerin, both resolution and contrast - wise. And, low index water soluble media are common.
} From Barron: The "sine law", non-fulfillment of which leads to spherical aberration and/or coma: n R (sinU) = n' I (sin e)
I could figure out the average thickness of my "too thick" mounts and accordingly prepare a medium with the optimum index to minimize SA. Problem is, the first "n" in the above equation represents the index of the medium (media!) between object and objective. I'll have up to 3 different indices of varying thicknesses (medium - glass - air)... I doubt one could use a weighted average... this won't be easy.
Dear Listservers: I know this topic has been hashed and rehashed in this forum. But the scanner business seems to be changing very quickly. I an interested in the best scanner in the $2000-3000 range for digitizing TEM negatives. I am working with materials so I would like a scanner that has sensitivity over a large contrast range. Any recent experiences would be highly appreciated.
Unfortunately several scanners in that price range that were suggested on the listserver have been discontinued (including the highly recommended UMAX). Thanks in advance.
Does any one has a used air-cooled water recirculator (for JEOL 100CX TEM) that you are willing to part with? We recently got a used TEM but it does not come with a water recirculator.
Sincerely, Ching-Hwa Kiang
-- ============================================================================== Dr. Ching-Hwa Kiang Visiting Assistant Professor Phone: (310) 206-0563 (Office) Cram Teacher-Scholar (310) 794-4124 (Lab) Department of Chemistry and Biochemistry (310) 206-4038 (Fax) University of California chk-at-chem.ucla.edu Los Angeles, CA 90095-1569 http://www.chem.ucla.edu/dept/Faculty/ ==============================================================================
Hello, we are doing some research on protective shields for x-ray radiation. does anyone know what is the permissible or background radiation intensity? also on refering to an old publication by the British Journal of Medicine, i found out that the maximum permissible dose(MPD) for exposure of the whole body to radiation expressed in rontgens is 0.3 per week, has this level changed or is it still the same? to calculate the MPD, we need to know the dose rate (in r min), does anyone have some info on this?
it would be great if you could give me some info on these topics.
Hello, we are doing some research on protective shields for x-ray radiation. does anyone know what is the permissible or background radiation intensity? also on refering to an old publication by the British Journal of Medicine, i found out that the maximum permissible dose(MPD) for exposure of the whole body to radiation expressed in rontgens is 0.3 per week, has this level changed or is it still the same? to calculate the MPD, we need to know the dose rate (in r min), does anyone have some info on this?
it would be great if you could give me some info on these topics.
I would like to know if anyone has an endmember calculation program for WDS } analysis of garnet. If so, how is the ferric and ferrous iron problem } resolved. My instrument setup is a 733 JEOL Microprobe with 5600/5500 } software. The program need not be an on line calculation, as I have the } setup in place to transferr the probe data to a PC for manipulation with a } Lotus spreadsheet. Hope to hear from someone soon.
Hi There! I've got an announcement about our latest workshop, so here it comes. Beware this is a long message ;-)
The UC Berkeley EM Lab, Ted Pella, Inc., and Leica present:
The 3rd Annual UC Berkeley Microwave Processing Techniques Workshop January 6-8, 1999
Breakthroughs in Microwave Processing: Techniques for Light and Electron Microscopy
Date and Times: January 6-8, 1999 9:00 am to 4:30 pm each day
Instructors: Rick Giberson, Ted Pella, Inc. Dr. Kent McDonald, Supervisor UC Berkeley EM Lab
Techinques covered:
1) Immunolabelling of microwave processed tissue for LM and EM.
2) Microwave-assisted immunolabelling on plastic, paraffin and cryoultramicrotomy sections.
3) Formalin fixation in the microwave-method and theory
4) Microwave-assisted processing of fresh tissue into paraffin
5) NEW microwave processing techniques for EM and LM
Who Should Attend:
Anyone interested in improving productivity in the lab. The course will examine the latest techniques in tissue processing for electron microscopy, formalin fixation and paraffin embedding of tissue using microwave technology. The course is designed for those individuals who want to solve problems and are receptive to new ideas that work. The theory and technique of microwave processing will be examined in detail.
Location:
Electron Microscopy Lab, 26 Giannini Hall, University of California, Berkeley
Workshop Particulars: Class size is limited to 15 participants
Cost: $400.00, which includes lunch every day and one group dinner
Type: Hands-on processing. You get to embed samples with the microwave ovens.
Contacts: Kathy Stangenberg or Rick Giberson at Ted Pella, Inc. Phone: 800-237-3526 Fax: 530-243-3761 E-mail: tedpel-at-aol.com
Hotel Accomodations:
A block of rooms at the Hotel Durant has been set aside, under the name "Ted Pella, Inc.", for attendees of the workshop, although you are not required to stay there. The hotel is within walking distance to the university. Individuals must reserve and pay all charges for their room. The single rate is $99 and the double rate is $110 with a cutt off date of 12/10/98. Check-in: 3:00 pm, check-out: 12:00 pm
For Reservations: Hotel Durant 2600 Durant Ave., Berkeley, CA 94704 Phone: 510-845-8981 or 800-238-7268 Fax: 510-486-8336 E-mail: durant-at-sfa.com
For a registration form see the Ted Pella, Inc., website at www.tedpella.com or phone the contact people at Ted Pella, Inc.
Well, there you go kids, your opportunity to see what I look like after reading all of my e-mails.
Hope to see you in January!
Paula :-)
p.s. I apologize for any typos I may have done. Check out the Ted Pella website, I think they have all the info I just wrote but in a better format.
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
I finally got on the list: my original question was posted before I was "officially" on. I apologize for any responses I may have missed yesterday or this morning.
Mark wrote: } } It is true that, for large angles of incidence, the spherical aberration of a plate is proportional to t/N (thickness over refractive index). So higher index might just give you a better image, if the other aberrations don't creep up on you, particularly longitudinal chromatic. But wouldn't you want a higher index cover slip as well? { {
Well, it now looks to me like Barbara was correct (no surprise) and that, for thick mounts, a low index would help; a high index would make it worse.( I sent a "correction on correction" post earlier today but it's not here somehow.) I had a heck of a time "ray - tracing" the situation, but I think that must be correct. Interesting idea about the higher index coverslip...
Bob wrote: } } I was wondering if it would be easier to find an objective with a correction collar on it? We have a 40x with a collar that we use for all our samples that are outside the standard thickness. It helps a lot. But I don't know if it would give enough resolution for your situation. { {
A correction collared objective would be nice. Maybe someday. But if adjustable tubelength does the same thing, then I'll have correction for any old objective. You folks with your fancy bino scopes ;-) would have a hard time doing that, although there should be other optical / mechanical methods of getting the same result.
Now that a low index medium seems to be what I need, does anyone have any info on; --- Glycerin Borate; once commercially available as "Aqua - Resin". Recommended by a well known expert. Low index by itself. I'm wondering if this was just a solution of Borax (Sodium Borate?) in glycerin or if it is a compound / molecule.
---- "Water Glass", a.k.a. Sodium Silicate. Essentially a water soluble "glass". Sounds like it might be moderately low n; maybe something could be added to further lower it if it's otherwise suitable.
We are looking for a way to set the contrast and the brightness values in the SEM's backscatter mode at the same level from day to day and rather so that any user can set it to this wanted level. We have not found a good way of doing this. Any suggestions?
Thanks a lot,
Tiina
************************************************************* Tiina Hallamaa, M.Sc (Eng) The Finnish Pulp and Paper Research Institute P.O.Box 70 FIN-02151 Espoo Finland ************************************************************* phone: +358-9-4371 492, fax: +358-9-464 305 email: Tiina.Hallamaa-at-kcl.fi
We use a waveform monitor on our AMRAY 1830. This allows us to set a pretty constant brightness and contrast level so that photographs will come out with the proper levels. This is much better than the auto brightness and contrast or small row of LED's that our SEM had as a factory meter. The waveform monitor resembles a small oscilloscope and has a fairly fine pitched grid on the screen. Your SEM manufacturer should be able to supply you with one. It has worked very well for us.
John Giles Principal Materials Engineer Honeywell Space Systems
} Dear all,
} We are looking for a way to set the contrast and the } brightness values in the SEM's backscatter mode at the } same level from day to day and rather so that any user } can set it to this wanted level. We have not found a } good way of doing this. Any suggestions?
Now I want to record the HRTEM images of Ceramic Grain Boundaries, but I found it is very difficult to find out a suitable sample area for both grains in the suitable orientations. Could you give me some advices about how to find them?
Many thanks,
--- Best Wishes,
JH Yu
Join 18 million Eudora users by signing up for a free Eudora Web-Mail account at http://www.eudoramail.com
If your SEM has a video waveform display available, it can be used with standards to consistently set BSE contrast (gain) and brightness (dc level). The best standards should be polished flat and the atomic number should bracket the anticipated BSE range required. Use the standards to set desired black/white levels.
Woody White McDermott Technology, Inc.
Dear all,
We are looking for a way to set the contrast and the brightness values in the SEM's backscatter mode at the same level from day to day and rather so that any user can set it to this wanted level. We have not found a good way of doing this. Any suggestions?
Thanks a lot,
Tiina
************************************************************* Tiina Hallamaa, M.Sc (Eng) The Finnish Pulp and Paper Research Institute P.O.Box 70 FIN-02151 Espoo Finland ************************************************************* phone: +358-9-4371 492, fax: +358-9-464 305 email: Tiina.Hallamaa-at-kcl.fi
Now I want to record the HRTEM images of Ceramic Grain Boundaries, but I found it is very difficult to find out a suitable sample area for both grains in the suitable orientations. Could you give me some advices about how to find them?
Many thanks,
--- Best Wishes,
JH Yu
Join 18 million Eudora users by signing up for a free Eudora Web-Mail account at http://www.eudoramail.com
First you should obtain NJDEP's Chapter 28 covering Radiation Protection Programs to see what rules need to be followed. New Jersey is very strict about this topic. The current edition 5/15/95 is good for five years.
A summary table in "7:28-6.1 Permissible dose rates, radiation levels and concentrations" indicates 1.25 rems per year. However, this section is long with many exceptions. If you send me an E-Mail with the details of your question, I will do my best to answer it.
J. Roy Nelson Material Testing Laboratory Pennington, NJ (609) 730-0575 jrnelson-at-nj1.aae.com
narahari ramanuja che stnt wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } we are doing some research on protective shields for x-ray } radiation. does anyone know what is the permissible or background } radiation intensity? also on refering to an old publication by the British } Journal of Medicine, i found out that the maximum permissible dose(MPD) } for exposure of the whole body to radiation expressed in rontgens is 0.3 } per week, has this level changed or is it still the same? to calculate the } MPD, we need to know the dose rate (in r min), does anyone have some info } on this? } } it would be great if you could give me some info on these topics. } } sincerely, } } Narahari. } } ________________________________________________________________________________ } } Narahari Ramanuja } } Office : Materials Synthesis & Characterization Lab, } Dept of Materials Science & Engineering, } NJIT, } ph # 201-596-3680 } email : nxr0308-at-megahertz.njit.edu } ________________________________________________________________________________ }
Tiina: SEM's have a linescan mode and this can be centred and modulated. The amplitude in the modulation then represents contrast and the position of the graph (which is centred before modulation) indicates brightness. This is easy enough to calibrate and teach to users. Modern instruments have various aids to obtain good exposures, some are simpler but they are never better than that modulation. Just fixing the brightness and contrast controls is rather more complicated, because this would necessitate fixing all working parameters in an SEM, always the same: specimens and coatings, WD, condenser setting and apertures, emission and specimen tilt. The list is endless, even the scintillator cannot age. Forget fixing just brightness and contrast amplifications and try to give users at least minimal instructions in SEM operations and that should include the modulated linescan. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Tuesday, December 01, 1998 4:47 PM, Tiina Hallamaa [SMTP:Tiina_Hallamaa-at-kcl.fi] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } Dear all, } } We are looking for a way to set the contrast and the } brightness values in } the SEM's backscatter mode at the same level from day to } day and rather so } that any user can set it to this wanted level. We have not } found a good } way of doing this. Any suggestions? } } Thanks a lot, } } } Tiina } } ********************************************************** } *** } Tiina Hallamaa, M.Sc (Eng) } The Finnish Pulp and Paper Research Institute } P.O.Box 70 } FIN-02151 Espoo } Finland } ********************************************************** } *** } phone: +358-9-4371 492, fax: +358-9-464 305 } email: Tiina.Hallamaa-at-kcl.fi } } } }
Just to expand on Woody's point, we use some TV tube coat (C) sprayed on a chip of calcite as a thin, portable standard to set the BSE brightness levels for concrete analyses. We simply lay the standard on the sample as we insert it into our microscope. Of course, one could also embed samples of brass or calcite or silica in epoxy and polish them flat to do the same thing. You could then have multiple reference levels.
Of course, some sort of waveform display is required. We have ours marked so that we can set the levels reproducibly.
Warren
At 07:56 AM 12/1/98 -0600, Woody wrote: } } Tiina, } } If your SEM has a video waveform display available, it can be used } with standards to consistently set BSE contrast (gain) and } brightness (dc level). The best standards should be polished flat } and the atomic number should bracket the anticipated BSE range } required. Use the standards to set desired black/white levels. } } Woody White } McDermott Technology, Inc. } } Dear all, } } We are looking for a way to set the contrast and the brightness values in } the SEM's backscatter mode at the same level from day to day and rather so } that any user can set it to this wanted level. We have not found a good } way of doing this. Any suggestions? } } Thanks a lot, } } Tiina
A colleague is interested in a standard to evaluate a variety of profilometry instruments. Both metallic and polymeric standards would be of use. X, Y, and Z directions would need to be measured, something in the 100-1000um range. Also, does anyone know a manufacturer who will make polystyrene latex spheres at a requested diameter?
TIA
======================= David Rose WL Gore and Associates 297 Blue Ball Road Elkton, MD 21921 =======================
Dear Narahari, The usual upper limit that is applied for any radiation-emitting devices in Canada and the USA is "0.5 mR/hr. at a distance of 5 cm from any part of the device". This is from the "Electron Microscopy Safety handbook" second edition, Barber and Mascorro editors. This is an excellent, slim, soft-cover book that should be in any EM lab, IMHO. There is a lot of other info on background radiation and other radiation hazards in the EM lab. You wrote: } Hello, } we are doing some research on protective shields for x-ray } radiation. does anyone know what is the permissible or background } radiation intensity? also on refering to an old publication by the British } Journal of Medicine, i found out that the maximum permissible dose(MPD) } for exposure of the whole body to radiation expressed in rontgens is 0.3 } per week, has this level changed or is it still the same? to calculate the } MPD, we need to know the dose rate (in r min), does anyone have some info } on this? } } it would be great if you could give me some info on these topics. } } sincerely, } } Narahari. } } ___________________________________________________________________________ _____ } } Narahari Ramanuja } } Office : Materials Synthesis & Characterization Lab, } Dept of Materials Science & Engineering, } NJIT, } ph # 201-596-3680 } email : nxr0308-at-megahertz.njit.edu } ___________________________________________________________________________ _____ } } } } } } } } } } } } } } } } } Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
I spent some time doing the same type of work in beta silicon nitride , and yesI found it very time consuming because one needs to find orientations such that the two grains are in orientations that can be imaged and at the same time the grain boundary needs to be parallel to the beam. You might find the paper by D.R. Clark (Ultramicroscopy 4 (1979)33-44) useful.
What I found to be helpful was to first check the image of the boundary at lower mags (typically X100K) in BF mode and tilt about an axis parallel to the image of the boundary until I attained a relatively sharp "minimum" in the width of the boundary .This is relatively fast and it allows you to eliminate those GBs that are way out of orientation ( I was using a 30 degree double tilt holder).
Once I had what seemed to be a suitable GB, then,at that point I would check the diffraction pattern to see if there were any Kikuchi lines (from either grain) that were parallel to the grain boundary and which corresponded to reflections that could be used for HRTEM imaging (i.e. large enough d-spacing). If that was the case, then I would tilt the sample along that Kikuchi line (i.e. maintaining that set of reflections for that particular grain) and at the same time check the DP from the other grain until a suitable orientation was attained for that second grain. More often than not, I had to move on to another location and start the whole operation again until all three elements(Grain 1, grain 2 and the GB) had the proper orientation.
In most instances my images consisted of two beam conditions but that was sufficient to resolve the GBs which were typically about 1 to 4 nm wide. In one afternoon session I could probably end up with three to five suitable boundaries (plus coffee breaks).
I hope this helps.
Jordi Marti
---------- } From: jiahui yu To: For sending TEM discussion -----------------------------------------------------------------------.
Dear Ceramic TEM experts and folks,
Now I want to record the HRTEM images of Ceramic Grain Boundaries, but I found it is very difficult to find out a suitable sample area for both grains in the suitable orientations. Could you give me some advices about how to find them?
Many thanks,
--- Best Wishes,
JH Yu
Join 18 million Eudora users by signing up for a free Eudora Web-Mail account at http://www.eudoramail.com
narahari ramanuja che stnt wrote: } } we are doing some research on protective shields for x-ray } radiation. does anyone know what is the permissible or background } radiation intensity? also on refering to an old publication by the British } Journal of Medicine, i found out that the maximum permissible dose(MPD) } for exposure of the whole body to radiation expressed in rontgens is 0.3 } per week, has this level changed or is it still the same? to calculate the } MPD, we need to know the dose rate (in r min), does anyone have some info } on this? } } it would be great if you could give me some info on these topics. } Dear Narahari, That must be a very old book. The current standards (at least in New York State) are 0.5 rem/year (about equal to 0.5 roentgens/year) for the general public and 5 rem/year for radiation workers; however, for women who are or may be pregnant, the lower level applies even for radiation workers. The dose rate must be measured with an ionization chamber detector (we use a Model 2588 from Nuclear Chicago). Due to the unevenness of shielding by the radiation source itself (e.g., the EM column), a calculation will not usually be useful. You should also operate the equipment to produce the worst possible radiation field when making these measurements--put the EM beam onto as many high-Z parts as possible, etc.--so that even operator mistakes will not result in exceeding the permissible levels. Good luck. Yours, Bill Tivol
A Philips EM400T w/ 6585 STEM was donated to our new EM teaching facility last year, to establish TEM & analytical capabilities (biology/engineering/physics/chemistry). On PEI's recommendation, we invested around $30K to move & refurbish the instrument, plus more to upgrade the EDS system.
End result: The column was in good shape to begin with, and now it is great (GO, Philips). However, after expending the greater part of the available funds in man-hours and parts on the STEM unit over a period of months, the STEM was merely transformed into a(n expensive!) heap of garbage that nothing can resuscitate (BOO, Philips).
Does anyone out in ListServerLand have a 6585 STEM unit you might loan or donate to our program?
PEI has offered to transport & install it for us (GO, Philips). Having put so much into our failed STEM, and remaining committed to the concept, we can also spend a little more for a(n operating!) unit.
Thanks, all.
Ann Hein Lehman TEM Lab Mgr, Trinity College LSC 314 300 Summit St. Hartford, CT 06106 860-297-4289 Ann.Lehman-at-exchange.cc.trincoll.edu
Our sputter coater (ISI PS-2) just died. I need parts for it and was hoping that someone may have an old one setting around that I could get for parts. I would also consider a used sputter coateer of any make that you may want to sell.
Thank you
Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
Tiina, } } There are a number of things you can do, depending on whether you want to } } a) just acquire images with the same contrast and brightness to facilitate } comparison or } b) want to acquire BSE images and get materials data from these. b) would } have to be split up further into } } i) image comparison } ii) semi-quantitative data without standard } iii) semi-quantitative with standard } } a) if you just want to get images with similar brightness and contrast, the } easiest and least expensive way would be to use the y-modulation or linescan } mode of the SEM, adjust brightness and contrast so that the amplitudes are } similar for different images, and take an image. } } b, i) if you want to use the Z-contrast of the SEM to obtain information } about material distribution in your sample, you probably have to pay more } attention to other parameters such as WD, beam current, contamination, etc. } Again, if you are just looking for a comparison of two areas you may be able } to use the method a). } } b,ii) Now it get's more interesting. If you want to obtain semi-quantitative } data you must quantify the gray levels of the image, which can then in turn } in principle be used to measure Z. If you don't want to use a standard, you } probably have to fix most of your parameters (WD, beam current, tilt angles, } etc). Once that is done, you need to acquire the images digitally, at which } point you can calibrate the intensity and then have the software threshold } different intensity levels and match them to different materials. The } matching could be done either by using theoretical values of I(z), or by } using values determined from known materials. } } b,iii) If you can use a standard in the SEM at the same time as the sample, } this would give you probably the best results. Acquire an image of the } standard, then of the sample under the same conditions. Software can then be } used to match the intensity of the standard to intensity on the sample and } obtain information about the material. Again, you could then use software to } threshold the intensity and tell you which material is where. } } I hope, that helped. If you have further questions regarding the digital } processing part, contact me through email. We have people in Finland who can } help you further. } } Michael Bode } } ******************************************************* } Disclaimer: } Soft Imaging System produces and sells image acquisition } and processing systems. We therefore have a vested } interest in some of the items mentioned above. } ******************************************************* } }
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS fax: (303) 234-9271 email: info-at-soft-imaging.com
} } } On Tuesday, December 01, 1998 4:47 PM, Tiina Hallamaa } [SMTP:Tiina_Hallamaa-at-kcl.fi] wrote: } } } ---------------------------------------------------------- } } -------------- } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } } ml } } } ---------------------------------------------------------- } } -------------. } } } } } } Dear all, } } } } We are looking for a way to set the contrast and the } } brightness values in } } the SEM's backscatter mode at the same level from day to } } day and rather so } } that any user can set it to this wanted level. We have } not } } found a good } } way of doing this. Any suggestions? } } } } Thanks a lot, } } } } } } Tiina } } } } } ********************************************************** } } *** } } Tiina Hallamaa, M.Sc (Eng) } } The Finnish Pulp and Paper Research Institute } } P.O.Box 70 } } FIN-02151 Espoo } } Finland } } } ********************************************************** } } *** } } phone: +358-9-4371 492, fax: +358-9-464 305 } } email: Tiina.Hallamaa-at-kcl.fi } } } } } } } } } } }
Does anyone have recommendations for a low-temperature lubricant for our old IEC cryostat. The literature that we have available just refers to "special low-temperature oil, product no. ####". We are thinking of molybdenum sulfide or graphite, but if there is any really good recommended lubricant for -30 degrees C, that's what we want.
Thanks in advance.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Having been off line for a while I see we have managed to confuse most of the world thanks to British Telecom. For Scott who asked about flat bottom vials we do manufacture these and much else besides. Stephen Griffiths from the Institute of Ophthalmology got it right although sadly there are no prizes for him this time. Anyone requiring details can contact us as follows:
TAAB Laboratories Equipment Ltd, 3 Minerva House, Calleva Park, Aldermaston Berkshire, RG7 8NA, England Tel ++44 (0) 118 981 7881 Fax 110 981 7881 Company e-mail sales-at-taab.co.uk
Randy: I agree that moly sulfide or graphite would be satisfactory. However, Apiezon N grease is designed for medium and very low temperature applications. It's sticky and a very thin film would stay in place for a long time. You can read about Apiezon N properties in our online by going to page M2 from the contents page. Disclaimer: ProSciTech is a supplier and sells this products. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
} } Hi, } } Does anyone have recommendations for a low-temperature } lubricant for our } old IEC cryostat. The literature that we have available } just refers to } "special low-temperature oil, product no. ####". We are } thinking of } molybdenum sulfide or graphite, but if there is any really } good recommended } lubricant for -30 degrees C, that's what we want. } } Thanks in advance. } } Randy } } } Randy Tindall } Electron Microscope Laboratory } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } } rtindell-at-nmsu.edu (work) } nrtindall-at-zianet.com (home)
Hi, We got ours from LIPSHAW and it is called LIPSHAW CRYOTOME LUBRICANT No. 291. The address Lipshaw Corp. 7446 Central Avenue Detroit. MI 48210. Hope this will help. Regards. Andre du Toit Department of Anatomical Pathology Tygerberg Hospital Cape Town South Africa.
A new web page was announced recently on this listserver; it has informative images and text that will help those who are doing educational outreach. It has just been expanded to include some difficult-to-access information:
The FOODS UNDER THE MICROSCOPE Web page (http://www.cyberus.ca/~scimat/) has been extended and now contains the TABLE OF CONTENTS of the FOOD STRUCTURE (formerly FOOD MICROSTRUCTURE) journal published in 1982-93. This particular page may be accessed separately at http://www.cyberus.ca/~scimat/journal.htm
Please send inquiries directly to Milos; he isn't a list subscriber.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
{!DOCTYPE HTML PUBLIC "-//W3C//DTD HTML 3.2//EN"} {HTML} {HEAD} {META HTTP-EQUIV="Content-Type" CONTENT="text/html; charset=us-ascii"} {META NAME="Generator" CONTENT="MS Exchange Server version 5.5.1960.3"} {TITLE} NiTi etchant {/TITLE} {/HEAD} {BODY}
{P} {FONT SIZE=2 FACE="Arial"} Any suggestion on proper etchant procedure to use in order to do NiTi Memory Shape alloys SEM images? {/FONT} {BR} {FONT SIZE=2 FACE="Arial"} Thank You in advance, Edoardo. {/FONT} {/P}
Does anybody know where I can get parts for a Sorvall Porter-Blum MT II ultramicrotome? If possible somewhere here in Canada, because of the exchange rate. I think there have been questions like this in the past, but I can't seem to access the archives. Thanks.
I'm not sure if graphite will work at those temperatures. Graphite gets its lubrication properties from the layers being intercalated with gas molecules. That is why it doesn't work in vacuum. MoS2 or WS2 works well at low temperatures for tribological applications. It also works well in vacuum applications. You probably will have to burnish it on and that could create some debris and there is debris that can occur during usage. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Randy Tindall To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hi,
Does anyone have recommendations for a low-temperature lubricant for our old IEC cryostat. The literature that we have available just refers to "special low-temperature oil, product no. ####". We are thinking of molybdenum sulfide or graphite, but if there is any really good recommended lubricant for -30 degrees C, that's what we want.
Thanks in advance.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Proper low-temperature cryostat oil is available from the major suppliers (i.e.Fisher), and is probably the best thing to use. My own experience is that other lubricants usually become impossibly thick at low temperatures.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Edoardo Bemporad } Any suggestion on proper etchant procedure to use in order to do NiTi Memory Shape alloys SEM images? Thank You in advance, Edoardo. {
I am looking to purchase several items. A cryostat, a paraffin prosessor, a freeze-substitution unit, and a freeze dryer. We need state-of-the-art equipment. Please contact me at one of the addresses below.
Thank you.
Kathy Walters / / Research Assistant III / /\ Center for Microscopy Research / /\ \ University of Iowa /_/ \ \ 85 EMRB ____ ((O)) Iowa City, Iowa 52242 | | / / || / / email: kwalters-at-emiris.iaf.uiowa.edu ----------- fax: (319)335-8049 ------------- www: http://www.uiowa.edu/~cemrf
Talk to me about joining the Iowa Microscopy Society.
Please respond to me directly, rather than on the listserve. Thank you - jpshield-at-arches.uga.edu
We are in the process of purchasing GFP filter(s) for Nikons (TE300 and Optiphot) we have at the Center. This is a multiuser facility, so naturally we sould like to have a good bit of versatility in the filter cube (if possible). Any suggestions or comments regarding wavelengths, etc... would be appreciated. Thanks ******************************************** John P. Shields Center for Ultrastructural Research Barrow Hall University of Georgia Athens, GA 30602 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
Hi, I'm looking for someone who might be interested in doing some work for me. I am looking for sulpher in some cell membranes. You may contact me at the following address if interested.
Kathy Walters / / Research Assistant III / /\ Center for Microscopy Research / /\ \ University of Iowa /_/ \ \ 85 EMRB ____ ((O)) Iowa City, Iowa 52242 | | / / || / / email: kwalters-at-emiris.iaf.uiowa.edu ----------- fax: (319)335-8049 ------------- www: http://www.uiowa.edu/~cemrf
Our beloved Fuji Pictrography 3000 has developed a minor glitch. At a reproducible spot in the image, there is a white (or almost white ) line traveling from top to bottom (in the direction of the short width on the 8 x 5" prints). You can only see it in the gray areas of the print and it doesn't appear to traverse solid black areas (we have moved a black bar into the middle of the spot and don't see it passing thru but it is on either side). This is not a straight line but looks like a squiggly line with an amplitude of a couple of millimeters (as if a pen vibrated across the surface as the paper was traveling under it). Has anybody experienced this problem? More importantly, has anybody solved it? Thanks in advance. Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Cryostat lubricants can be found in the Fisher and VWR catalogs in the Histology sections
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
Who manufacturesand/or markets the Centaurus BSE detector?
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
First you should obtain NJDEP's Chapter 28 covering Radiation Protection Programs to see what rules need to be followed. New Jersey is very strict about this topic. The current edition 5/15/95 is good for five years.
A summary table in "7:28-6.1 Permissible dose rates, radiation levels and concentrations" indicates 1.25 rems per year. However, this section is long with many exceptions. If you send me an E-Mail with the details of your question, I will do my best to answer it.
J. Roy Nelson Material Testing Laboratory Pennington, NJ (609) 730-0575 jrnelson-at-nj1.aae.com
narahari ramanuja che stnt wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } we are doing some research on protective shields for x-ray } radiation. does anyone know what is the permissible or background } radiation intensity? also on refering to an old publication by the British } Journal of Medicine, i found out that the maximum permissible dose(MPD) } for exposure of the whole body to radiation expressed in rontgens is 0.3 } per week, has this level changed or is it still the same? to calculate the } MPD, we need to know the dose rate (in r min), does anyone have some info } on this? } } it would be great if you could give me some info on these topics. } } sincerely, } } Narahari. } } ________________________________________________________________________________ } } Narahari Ramanuja } } Office : Materials Synthesis & Characterization Lab, } Dept of Materials Science & Engineering, } NJIT, } ph # 201-596-3680 } email : nxr0308-at-megahertz.njit.edu } ________________________________________________________________________________ }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
I forwarded some recent postings on negative staining for bacteriophages to a colleague. He is now interested in some details on the techniques for glow discharge and on the benefits or drawbacks of using uranyl formiate instead of uranyl acetate. If anyone could post some references or personal experiences, both he and I would be grateful.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
12/3/98 10:34 AM
From a Leco Corp. metallography book-1977 3000 lakeview Ave., St. Joseph, Michigan, 49085
#50 5ml acetic acid Electolytic, platinum touch wire at 1.5 volts 10ml nitric acid for 20 to 60 seconds 85ml water
# 133 50ml nitric acid Mix fresh. Swab 5 to 30 seconds or electolytic at 50ml acetic acid 5-10 volts, for 5 to 60 seconds. (Polishes at high currents).
#143 0.01-1 g Allow mixture to age a few minutes, then swab or chromium trioxide immerse a few seconds to a few minutes. 100ml HCL
# 151 10ml HF Swab 5 to 30 seconds 25ml nitric acid (Caution: Wear gloves rated for HF acid 150ml water protection-and use a fume hood).
Note: I have not personally used any of these etches but hope the information helps Edoardo Bemporad and others.
Bernard Kestel Materials Science Division, Bldg. 212 Argonne National Laboratory 9700 South Cass Avenue Argonne Illinois, 60439
E-mail: {kestel-at-anl.gov} FAX: (630) 252-4289
--------------------------------------
Any suggestion on proper etchant procedure to use in order to do NiTi Memory Shape alloys SEM images? Thank You in advance, Edoardo.
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{P} {FONT SIZE=2 FACE="Arial"} Any suggestion on proper etchant procedure to use in order to do NiTi Memory Shape alloys SEM images? {/FONT} {BR} {FONT SIZE=2 FACE="Arial"} Thank You in advance, Edoardo. {/FONT} {/P}
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Would someone know if there is antibody to cy3 and who carries it. I know that there are few companies that sell antibodies for FITC. Internet search gives me all the cy3 conjugates. Thank you, Lilith --------------------- Lilith Ohannessian-Barry NRC, IBS, Ottawa, Ont. K1A 0R6 CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
} } Hello all, } } I forwarded some recent postings on negative staining for bacteriophages to } a colleague. He is now interested in some details on the techniques for } glow discharge and on the benefits or drawbacks of using uranyl formiate } instead of uranyl acetate. If anyone could post some references or personal } experiences, both he and I would be grateful. } } Thank you. } } Kim
The original thread on negative staining bacteriophages was in response to a query I made. I am appending to this message a slightly condensed summary of all the replies. I haven't tried all the suggestions yet but I did get much better staining from Ammonium Hydroxide neutralized PTA. The Am Molybdate was different and needs further examination. I plan to try additional suggestions as soon as I get a fresh phage prep. The advice once again proved how useful this listserver is to me. Thanks to everybody who shared their experiences. Tom
In doing Bacteria, we found it advantagous to dilute the stain 1/30 with water. This made the Bacteria show up with good inner structure instead of "the blob".
Perhaps a more dilute solution may stain a little less, but show up fine detail better? If that doesn't work... more concentrated? ** Also did you fix the pacterophages? Fixing may help *************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine U of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
a) Did you 'glow discharge' the grid shortly before you adsorbed the drop? b) Have you tried Uranyl Formiate solution? (I think 0.75%w/w and pH around 4 should be good.) c) Have you tried lower concentration of Uranyl salt? Maybe the fine details are embedded in a thick salt layer so that you don't get any contrast there.
The procedure I normally use is: - prepare parafilm with at least 2 droplets of water (or sometimes buffer?) and another two of stain solution - glow discharge the grid for around 10 to 20 s (blue-white) - clamp it into tweezer - put 5um drop on the grid and wait... whatever.. 30 s, 60 s.... - touch with filter paper to soak solution away (blotting) - wash on first drop of water by touching the surface, blot again - repeat above step at least once - then go to stain droplet and do the same - the second stain droplet is then used to do the staining procedure, so touch the surface and wait for a few seconds (lets say 10 to 20) - blot a last time and that's it
You may not need all these steps, but I have been very successful with this technique and don't see any reason not to do it this way. If you don't have Uranyl Formiate, you can try the upper protocol with Uranyl Acetate (2% or lower). My experience is that usually it should work as nicely as UFo, but I was told that UAc does 'melt' more strongly and more quickly in the electron beam. I am looking forward to other recipies. I am very sure that you can find as many recipies as there are microscopists...
By the way, I have never been successful using PTA. Even with very sensitive proteins which don't like a low pH, I always had the best results in using Uranyl solutions.
Bettina Wolpensinger Electron Microscope Unit University of New South Wales Sydney, NSW 2052, AUSTRALIA b.wolpensinger-at-unsw.edu.au http://srv.emunit.unsw.edu.au
I'm no expert but we have had problems because of the following:
My experience with UA is that when it works it's great but it doesn't always work.
With PTA a bad batch of PTA stock could result in poor images - especially if it's old or not really a good quality reagent..
Do you adjust the pH of your PTA? It is normally fairly acid and should be adjusted to between 6 and 7 but you can experiment. We have always understood that it is best to use potassium hydroxide and not sodium hydroxide to adjust - personally I've seen little difference.
If you are using carbon coatings on your grids and clean preparations of virus maybe you need a little BSA to help spreading (not normally a problem with our samples).
If none of the above work it might be worth trying some of the newer more expensive stains like sodium silicotungstate or methylamine tungstate which can give nice results.
I suspect you may have tried most of the above, but I hope that something will help. Good Luck.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Bldg University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk _____________________________________________________________________
Several years a go our lab switched from the traditional KPTA to NHPTA. which enhances the fine structural details often lost with more aggressive stains. Prepare a 1% solution of phosphotungstic acid , then pH the stain with 1N ammonium hydroxide to 6.5 and / or 7.0. Regards, Skip } From: L R MELSEN {lmelsen-at-emory.edu} _____________________________________________________________________ When you stain (this is for t4 and UAC) allow four to eight drops of stain I use, almost as routine, ammonium molibdate, 1 or 2% in water with very good results (for visualize African swine fever virus, herpesvirus, rotaviruses, polyomaviruses, adenoviruses). My protocol: (1) a drop of the virus containing pellet over a carbon coated grid; (2) wash in situ with 5-7 drops of negative stain; (3) dry with paper filter.
J.F.Moura Nunes Lab. Electron Microscopy Portuguese Cancer Institute 1093 Lisbon Codex Portugal
_____________________________________________________________________ When you stain (this is for t4 and UAC) allow four to eight drops of stain to drip on and off the grid. This works well if you hold the grid at a 45 degree angle. If you wash try 0.1mNaCl. Water can osmotically shock the specimen. Bob BOB-at-befvax.uchicago.edu
____________________________________________________________________ I would only add to Paul Webster's and other's comments that an amorphous carbon film that has been glow-discharged in a partial vacuum with air for about 60 seconds is pretty hard to beat when it comes to a substrate for negative staining. We used this procedure for T4 on one of my (numerous) postdocs in the Dept. of Microbiology at the Biozentrum in Basel. We never found anything to beat it. The stain spreads beautifully on the surface.
Robert (Bob) Chiovetti Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com
_____________________________________________________________________ In addition to the many recipes for stains you must be receiving, you might also like to try a simple "low-dose" imaging protocol. I have found when using negative stains that the image selection and focussing procedures can damage fragile structures in the sample. Focussing on an area close to a particle and then moving over to take the picture can help prevent this. If you own a newer TEM, you might even have a low dose capability built in. I have found this method very useful for small, fragile samples, even when coated with thick films of aqueous uranyl acetate or phosphotungstic acid.
Alternatively, the support film might be too thick to image the fine detail. Is your grid carbon coated, as you post, or is it formvar/carbon coated, which is more usual? Carbon alone offers more for high resolution work. Finally, have you checked out the vibrations affecting the microscope? High resolution requires stable conditions. Check out my web site for a brief summary of the options {http://www.hei.org/htm/neg.htm} . I look forward to postings containing stain receipies.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
_____________________________________________________________________ Try 3% ammonium molybdate pH 7.2 , I have good results with it.
Ann Fook Yang EM Unit Eastern Cereal and Oilseed Research Centre Agriculture and Agri-Food Canada 960 Carling Ave Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Tel.: 613-759-1638 Fax: 613-759-1701
e-mail:yanga-at-em.agr.ca
_____________________________________________________________________ Look up work by this expert:
Ackermann HW. Krisch HM. Felix d'Herelle Reference Center for Bacterial Viruses, Department of Microbiology, Faculty of Medicine, Laval University, Sainte-Foy, Canada. A catalogue of T4-type bacteriophages. [Review] [115 refs] Archives of Virology. 142(12):2329-45, 1997.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735 Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax) --============_-1299405738==_ma============ Content-Type: text/enriched; charset="us-ascii"
}
} Hello all,
}
} I forwarded some recent postings on negative staining for bacteriophages to
} a colleague. He is now interested in some details on the techniques for
} glow discharge and on the benefits or drawbacks of using uranyl formiate
} instead of uranyl acetate. If anyone could post some references or personal
} experiences, both he and I would be grateful.
}
} Thank you.
}
} Kim
The original thread on negative staining bacteriophages was in response to a query I made. I am appending to this message a slightly condensed summary of all the replies. I haven't tried all the suggestions yet but I did get much better staining from Ammonium Hydroxide neutralized PTA. The Am Molybdate was different and needs further examination. I plan to try additional suggestions as soon as I get a fresh phage prep. The advice once again proved how useful this listserver is to me. Thanks to everybody who shared their experiences. Tom
In doing Bacteria, we found it advantagous to dilute the stain 1/30 with water. This made the Bacteria show up with good inner structure instead of "the blob".
Perhaps a more dilute solution may stain a little less, but show up fine detail better? If that doesn't work... more concentrated? ** Also did you fix the pacterophages? Fixing may help
***************************
Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine U of Illinois
2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
a) Did you 'glow discharge' the grid shortly before you adsorbed the drop?
b) Have you tried Uranyl Formiate solution? (I think 0.75%w/w and pH around 4 should be good.)
c) Have you tried lower concentration of Uranyl salt? Maybe the fine details are embedded in a thick salt layer so that you don't get any contrast there.
The procedure I normally use is:
- prepare parafilm with at least 2 droplets of water (or sometimes buffer?) and
another two of stain solution
- glow discharge the grid for around 10 to 20 s (blue-white)
- clamp it into tweezer
- put 5um drop on the grid and wait... whatever.. 30 s, 60 s....
- touch with filter paper to soak solution away (blotting)
- wash on first drop of water by touching the surface, blot again
- repeat above step at least once
- then go to stain droplet and do the same
- the second stain droplet is then used to do the staining procedure, so touch
the surface and wait for a few seconds (lets say 10 to 20)
- blot a last time and that's it
You may not need all these steps, but I have been very successful with this technique and don't see any reason not to do it this way. If you don't have Uranyl Formiate, you can try the upper protocol with Uranyl Acetate (2% or lower). My experience is that usually it should work as nicely as UFo, but I was told that UAc does 'melt' more strongly and more quickly in the electron beam. I am looking forward to other recipies. I am very sure that you can find as many recipies as there are microscopists...
By the way, I have never been successful using PTA. Even with very sensitive proteins which don't like a low pH, I always had the best results in using Uranyl solutions.
Bettina Wolpensinger Electron Microscope Unit University of New South Wales
Sydney, NSW 2052, AUSTRALIA b.wolpensinger-at-unsw.edu.au http://srv.emunit.unsw.edu.au
I'm no expert but we have had problems because of the following:
My experience with UA is that when it works it's great but it doesn't always work.
With PTA a bad batch of PTA stock could result in poor images - especially if it's old or not really a good quality reagent..
Do you adjust the pH of your PTA? It is normally fairly acid and should be
adjusted to between 6 and 7 but you can experiment. We have always understood that it is best to use potassium hydroxide and not sodium hydroxide to adjust - personally I've seen little difference.
If you are using carbon coatings on your grids and clean preparations of virus maybe you need a little BSA to help spreading (not normally a problem with our samples).
If none of the above work it might be worth trying some of the newer more expensive stains like sodium silicotungstate or methylamine tungstate which can give nice results.
I suspect you may have tried most of the above, but I hope that something will help. Good Luck.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Bldg
University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
Several years a go our lab switched from the traditional KPTA to NHPTA. which enhances the fine structural details often lost with more aggressive stains. Prepare a 1% solution of phosphotungstic acid , then pH the stain with 1N ammonium hydroxide to 6.5 and / or 7.0.
When you stain (this is for t4 and UAC) allow four to eight drops of stain I use, almost as routine, ammonium molibdate, 1 or 2% in water with very good results (for visualize African swine fever virus, herpesvirus, rotaviruses, polyomaviruses, adenoviruses). My protocol: (1) a drop of the virus containing pellet over a carbon coated grid; (2) wash in situ with 5-7 drops of negative stain; (3) dry with paper filter.
J.F.Moura Nunes Lab. Electron Microscopy
Portuguese Cancer Institute 1093 Lisbon Codex Portugal
When you stain (this is for t4 and UAC) allow four to eight drops of stain to drip on and off the grid. This works well if you hold the grid at a 45 degree angle. If you wash try 0.1mNaCl. Water can osmotically shock the specimen.
In addition to the many recipes for stains you must be receiving, you might also like to try a simple "low-dose" imaging protocol. I have found when using negative stains that the image selection and focussing procedures can damage fragile structures in the sample. Focussing on an area close to a particle and then moving over to take the picture can help prevent this. If you own a newer TEM, you might even have a low dose capability built in. I have found this method very useful for small, fragile samples, even when coated with thick films of aqueous uranyl acetate or phosphotungstic acid.
Alternatively, the support film might be too thick to image the fine detail. Is your grid carbon coated, as you post, or is it formvar/carbon coated, which is more usual? Carbon alone offers more for high resolution work. Finally, have you checked out the vibrations affecting the microscope? High resolution requires stable conditions. Check out my web site for a brief summary of the options { {http://www.hei.org/htm/neg.htm} . I look forward to postings containing stain receipies.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I understand that there is = information on=20 scanning and digital cameras at this address. Is that true and how do I = access=20 it {/FONT} {/DIV} {/BODY} {/HTML}
Thanks to all who replied. For those of you who might want the information, the Centaurus back-scattered electron detector can be found in the USA at:
http://www.gwelectronics.com/backed.htm
GW Electronics Inc. 6981 Peachtree Industrial Blvd. Norcross, GA 30092-3601 Tel: 770-449-0707 800-325-5556
Mark Massey of EDAX Europe told me that they are manufactured by KE Developments in the UK: K. E. Development Limited The Mount, Toft Cambridge CB3 7RL UK Tel: +44 1223 263 948 Fax: +44 1223 263 532
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
NiTi alloys are very sensitive to structural damage during polishing. Be sure to use care in mechanical polishing or use electro-polishing.
Electro-polish: 20% H2SO4 in MeOH 28-29v for 1-5 min.
Immersion etch: similar to ASTM etch No. 151 and ASTM etch No. 209 14.1ml HNO3 3.2ml HF 82.7ml H2O submerge for 10-15 sec. with gentile agitation.
Good luck Sam. O. Mancuso Group Leader, Physical-Mechanical Metallurgy Special Metals Corporation 4317 Middle Settlement Road New Hartford, New York 13413 U.S.A. {} {} {} Phone: (315) 798-2920 Fax: (315) 798-2001 email: mancuso4-at-ix.netcom.com
-----Original Message----- } From: Edoardo Bemporad [mailto:e.bemporad-at-materials10.dimi.uniroma3.it] Sent: Wednesday, December 02, 1998 9:56 AM To: microscopy-at-sparc5.microscopy.com
Dear Kim: An excellent reference on glow discharging is in Advances in Optical and Electron Microscopy 8:107-135 (1982) by Dubochet, Groom and Mueller- Neuteboom. Don Gantz Boston Univ Med School gantz-at-med-biophd.bu.edu
OK, well I have really done it now - bought a whole TEM sectioning workstation in order to get the glass knife breaker. I am an LM type who is starting to use glass knives for some of my sectioning work (that is another story). Anyway, this has created two problems:
1. I need a copy of the manual for an LKB 7801-B knife breaker so I can get going with this project. I think I remember how from my grad school days, but would be a lot more confident with a copy of the manual. Would be happy to pay for the copying and postage charges.
2. I am left with an LKB Ultrotome III in what is reported to be good working condition - this is surplus to my rather simple needs (I am currently doing my sectioning on a nice old Lipshaw microtome). The microtome consists of the mail unit (8800), the control unit (8802-A), the base (8809-A), and the illuminator base for the sterio microscope head (no sterio microscope of course). Again, I have not used this unit, so I can not vouch for it's mechanical/operating condition - it is clean and apparently complete. I would be open to reasonable offers - biased in favor of someone in the Washington DC - Baltimore, MD area. This thing is HEAVY, and I would have to be pretty well compensated to want to build a proper crate for it. It would be much easier for someone to pick it up, or at least if I could arrange to meet someone half way. As they say, money is always nice, but interesting trades for LM equipment are also quite possible (my lab is based on old Leitz 170mm Ortholux and Laborlux scopes, and a variety of AO and B&L sterio scopes - those I need more of always).
Thanks for any assistance.
Stephen Poe - I can also be reached at my at-home E-mail address spoefish-at-mindspring.com
The Centaurus BSE detector is marketed by GW Electronics, (770)449-0707.
Best regards,
Angela
} Who manufacturesand/or markets the Centaurus BSE detector? } } Russell E. Cook } Electron Microscopy Center for Materials Research } Argonne National Laboratory } Building 212 } 9700 South Cass Avenue } Argonne, IL 60439 } (630)252-7194 } FAX: (630)252-4798 } --------------------------------------------- Angela V. Klaus
Manager - Core Microscopy Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
OK, well I have really done it now - bought a whole TEM sectioning workstation in order to get the glass knife breaker. I am an LM type who is starting to use glass knives for some of my sectioning work (that is another story). Anyway, this has created two problems:
1. I need a copy of the manual for an LKB 7801-B knife breaker so I can get going with this project. I think I remember how from my grad school days, but would be a lot more confident with a copy of the manual. Would be happy to pay for the copying and postage charges.
2. I am left with an LKB Ultrotome III in what is reported to be good working condition - this is surplus to my rather simple needs (I am currently doing my sectioning on a nice old Lipshaw microtome). The microtome consists of the mail unit (8800), the control unit (8802-A), the base (8809-A), and the illuminator base for the sterio microscope head (no sterio microscope of course). Again, I have not used this unit, so I can not vouch for it's mechanical/operating condition - it is clean and apparently complete. I would be open to reasonable offers - biased in favor of someone in the Washington DC - Baltimore, MD area. This thing is HEAVY, and I would have to be pretty well compensated to want to build a proper crate for it. It would be much easier for someone to pick it up, or at least if I could arrange to meet someone half way. As they say, money is always nice, but interesting trades for LM equipment are also quite possible (my lab is based on old Leitz 170mm Ortholux and Laborlux scopes, and a variety of AO and B&L sterio scopes - those I need more of always).
Thanks for any assistance.
Stephen Poe - I can also be reached at my at-home E-mail address spoefish-at-mindspring.com
OK, well I have really done it now - bought a whole TEM sectioning workstation in order to get the glass knife breaker. I am an LM type who is starting to use glass knives for some of my sectioning work (that is another story). Anyway, this has created two problems:
1. I need a copy of the manual for an LKB 7801-B knife breaker so I can get going with this project. I think I remember how from my grad school days, but would be a lot more confident with a copy of the manual. Would be happy to pay for the copying and postage charges.
2. I am left with an LKB Ultrotome III in what is reported to be good working condition - this is surplus to my rather simple needs (I am currently doing my sectioning on a nice old Lipshaw microtome). The microtome consists of the mail unit (8800), the control unit (8802-A), the base (8809-A), and the illuminator base for the sterio microscope head (no sterio microscope of course). Again, I have not used this unit, so I can not vouch for it's mechanical/operating condition - it is clean and apparently complete. I would be open to reasonable offers - biased in favor of someone in the Washington DC - Baltimore, MD area. This thing is HEAVY, and I would have to be pretty well compensated to want to build a proper crate for it. It would be much easier for someone to pick it up, or at least if I could arrange to meet someone half way. As they say, money is always nice, but interesting trades for LM equipment are also quite possible (my lab is based on old Leitz 170mm Ortholux and Laborlux scopes, and a variety of AO and B&L sterio scopes - those I need more of always).
Thanks for any assistance.
Stephen Poe - I can also be reached at my at-home E-mail address spoefish-at-mindspring.com
I've been batting my head against my Fischione Jet Polisher trying to figure this one out! Please help if you can. My original material is full of lovely voids and pits, Si particles are around 10 microns, and at a thickness of 150 to 200 microns I have experienced some holes that go all the way through the material. If I try initial thinning of the sample on less than 500 grit SiC paper I get even more particle pullout...Any ideas?
} Our sputter coater (ISI PS-2) just died. I need parts for it and was } hoping that someone may have an old one setting around that I could } get for parts. I would also consider a used sputter coater of any } make that you may want to sell.
} Thank you
} Marty Reed } Equipment Technician } Biology Department } Humboldt State University
Dear Marty,
The ISI PS-2 sputter coater was made by us way back in the early 1970s during our days as Polaron Equipment Ltd. Although the PS-2 has not been manufactured for many years we do still carry some spare parts. For further information please contact our local distributor, Energy Beam Sciences Inc. EBS ebs-at-ebsciences.com tel: 413 786 9322
Please note our new address, telephone and fax number for any enquiries concerning the Polaron range
Best regards Mike Wombwell Polaron range Business Manager V G Microtech The Birches Industrial Estate Imberhorne Lane East Grinstead West Sussex RH19 1UB UK Direct line: +44 (0)1342 310296 Switchboard: +44 (0)1342 327211 Fax: +44 (0)1342 315074 http://www.vgmicrotech.com/polaron-range E&OE
Dear Dorrance Before I will be able to suggest some advice, I would need more information.
} My original material is full of } lovely voids and pits, Si particles are around 10 microns, and at a } thickness of 150 to 200 microns I have experienced some holes that go all } the way through the material.
Firstly what is your original material? That will determine quite a few tings Eg. rate of thinning vs Si particle thinning rate.
} If I try initial thinning of the sample on } less than 500 grit SiC paper I get even more particle pullout...Any ideas? How is your sample "hold' in order to polish. Is it already 3mm diameter or will you punch the disks later. what lubrication do you use if any. } } Thanks in advance for any help. Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
We are considering looking for a Wavelength-Dispersive Spectrometer to improve our detection of sodium in biological samples, reasoning that the better energy resolution of the detector will result in narrower, taller peaks with the same background level, yielding improved peak-to-background ratios.
Our samples are frozen hydrated and dried biological tissues; we would like to be able to differentiate 50mMolar from 150mMolar sodium (0.25 and .75% by weight in hydrated samples).
Our present EDS detector has a semi-thin window and is doing better than average for sodium and we would prefer not going to an ultrathin window or windowless detector.
Might we expect a significant improvement with WDS?
Does anyone know of a spectrometer that might be donated?
Your comments and suggestions will be appreciated.
Thanks,
Jacob
Jacob Bastacky, M.D. Room 116 Donner Laboratory Lawrence Berkeley Laboratory University of California Berkeley, California 94720 Telephones: 510.486.4606 office, 510.486.4605 lab, 510.845.8031 fax email: sjbastacky-at-lbl.gov
I am sure that I am not the only one who runs into problems with color representation when printing out fluorescence (RGB) images on paper. I think that I am aware of the cause for the problem, but I got no final solution for it.
While fluorescence images (and other images representing brightness values as pure red, green or blue colors) look pretty well on the monitor or when presented as a slide or with a laser beamer (working in RGB mode), the translation of channels into colors renders pure colors and most color combinations too dark on the printout. This color translation has, in principle, nothing to do with the translation of RGB colors into YMCK printer colors by the printer driver, but is rather a consequence of a one-to-one translation of channels into pure colors. For instance, printing pixels representing bright green fluorescence (e.g. the maximum brightness value of 255 at 8 bit resolution per channel) results in a saturated green (maximum green value) on the printout, which appears too dark to the eye. Remembering bright green fluorescence in the microscope, our eyes await to see a bright green color (white/green) on the printout, which may be represented by a certain mixture of green, red and blue (or the respective mixture of CMYK printer colors).
I think what would help would be a good look-up table or a set of tables (or, e.g., a Photoshop macro) to represent fluorescence images (single channels and channel combinations) appropriately on different color printer types (laser, inkjet and thermotransfer/thermoosublimation printers). Experimenting with Photoshop (and other software's) parameters (brightness and saturation corrections etc.) did not result in a color representation I think (and dream) of, and is error-prone.
I am aware of the fact that, in general, printers cannot represent the whole RGB color space (especially green colors), but I think that good look-up tables or macros could help a lot.
Thanks in advance
Guenter
---------------------------------- Dr. Guenter Giese MPI fuer Medizinische Forschung Jahnstr. 29 D-69120 Heidelberg Phone (Germany or 0-)6221-486-320 Fax (Germany or 0-)6221-486-325 e-mail: ggiese-at-mzf.mpimf-heidelberg.mpg.de ------------------------------------------
Tong, If you can use tungsten for your application, McCrone Accessories and Components sells pre-made probes and will also sell you tungsten wire and sodium nitrite so you can etch your own wire down to the desired size. No financial interest in McCrone, just a satisfied customer.
Vern Rieck Quality Engineer Aavid Thermal Products, Inc. 603-629-2224 direct 603-666-4100 main fax 603-669-2044 rieck-at-aavid.com
-----Original Message----- From: Tong Wang [SMTP:tong-at-jlab.org] Sent: Thursday, December 03, 1998 11:50 AM To: microscopy-at-sparc5.microscopy.com Subject: fine metal tip used in microscopy
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Jacob Bastacky wrote: } } We are considering looking for a Wavelength-Dispersive Spectrometer to } improve our detection of sodium in biological samples, reasoning that the } better energy resolution of the detector will result in narrower, taller } peaks with the same background level, yielding improved peak-to-background } ratios.
This is correct. } } Our samples are frozen hydrated and dried biological tissues; we would like } to be able to differentiate 50mMolar from 150mMolar sodium (0.25 and .75% } by weight in hydrated samples). } This should be pretty straight-forward. } } Might we expect a significant improvement with WDS? } Yes.
} Does anyone know of a spectrometer that might be donated?
One permanant media that we have used is Molecular Probes "Pro-Long Anti-Fade. It dries hard and we store them at room temp with our histochemically stained specimens. I don't know if I would call it totally permanant. Samples still do fade with time and exposure.
Bob
On Thu, 3 Dec 1998, Amanda Harman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would be grateful if anyone could tell me if there is a fluorescent } mounting media that is permanent? } Amanda Harman. } } } }
This is a basic question for the materials scientists. I am interested in finding a simple reference which might delineate the different types of stainless steel, including relative hardness, tensile strength, etc. I'm also interested in seeing the same information plus the electrical characteristics of the various alloys of indium. In this case, melting points are of interest, and the exact formulae of "Wood's metal" and another that we use called "Cerralloy". Many thanks- Grace
IUMAS 2000, The 2000 Meeting of the International Union of Microbeam=20 Analysis Societies, will be held at The King Kamehameha Hotel in=20 Kailua-Kona, HawaiiJuly 9th-13th 2000=00.
This meeting is jointly sponsored by MAS, the European MAS, The Japanese=20 MAS, the Chinese MAS, the Australian MAS and the Korean MAS. Check out the= =20 information at:
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Hi Dorrance,
Silicon Carbide abrasive is one of your main problems. You need to use diamond for thinning. Silicon carbide does not cut silicon very well, in fact it will fracture it and that is why you are getting pullout. Try diamond lapping film with some glycol as the lubricant.
Good Luck,
Gary Liechty Allied High Tech Products, Inc.
McLean, Dorrance wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All, } } I've been batting my head against my Fischione Jet Polisher trying to figure } this one out! Please help if you can. My original material is full of } lovely voids and pits, Si particles are around 10 microns, and at a } thickness of 150 to 200 microns I have experienced some holes that go all } the way through the material. If I try initial thinning of the sample on } less than 500 grit SiC paper I get even more particle pullout...Any ideas? } } Thanks in advance for any help. } } Dorrance
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I am evaluating FEG semi-in-lens SEMs for possible purchase. I am interested in BSE imaging at low acc. voltages ( {3kV). I would like to get some feedback as to people's experiences with using BSE mode at low voltages. Specifically, what kind of detectors are being used and what kind of results? What is the lowest acc. voltage that you can reasonably use and still get good results. Thanks,
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Hello Grace,
Contact ASM International in Ohio, 800-336-5152 for infomation on books that will have what you are seeking.
Yours Truly,
Gary Liechty Allied High Tech Products, Inc.
Grace Kennedy wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } This is a basic question for the materials scientists. I am interested in } finding a simple reference which might delineate the different types of } stainless steel, including relative hardness, tensile strength, etc. I'm } also interested in seeing the same information plus the electrical } characteristics of the various alloys of indium. In this case, melting } points are of interest, and the exact formulae of "Wood's metal" and } another that we use called "Cerralloy". Many thanks- Grace
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Dear Jacob, The improvement of the WDX peak-to-background ratio over the EDX is even more because the background is much lower. The detection limit is much lower, so it is easier to quantify levels such as yours, which are close to the EDX detection level. The problems of WDX are first: high cost and second:, the need for a flat sample with exact geometry to the electron beam. You must also run standards that are exactly the same geometry. The WDX is much fussier than EDX on sample geometry and the analyses generally take much longer, but have higher precision and lower detection level. I have no experience with biological samples on the WDX, but there is a listserver for microprobe analysis, i.e. WDX analysis. To subscribe to the microprobe listserver use: microprobe-at-www.microanalysis.org, to post to it use: microprobe-at-microanalysis.org. I don't know of any WDX spectrometers for donation, there aren't that many around and they have to fit your SEM model.
You wrote: } } We are considering looking for a Wavelength-Dispersive Spectrometer to } improve our detection of sodium in biological samples, reasoning that the } better energy resolution of the detector will result in narrower, taller } peaks with the same background level, yielding improved peak-to-background } ratios. } } Our samples are frozen hydrated and dried biological tissues; we would like } to be able to differentiate 50mMolar from 150mMolar sodium (0.25 and .75% } by weight in hydrated samples). } } Our present EDS detector has a semi-thin window and is doing better than } average for sodium and we would prefer not going to an ultrathin window or } windowless detector. } } Might we expect a significant improvement with WDS? } } Does anyone know of a spectrometer that might be donated? } } Your comments and suggestions will be appreciated. } } Thanks, } } Jacob } } Jacob Bastacky, M.D.
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Hi Listserver! I am doing some immunohistochemistry and have made two attempts but with no luck at getting any gold label. My question is whether to use a lower dilution gold-antibody or extend the incubation from 4 hours to overnight or both? I am a self-confessed rookie at this, so any information on the subject would be appreciated .
Position Available University of Central Florida Advanced Materials Processing and Analysis Center (AMPAC)
The Advanced Materials Processing and Analysis Center (AMPAC) is seeking candidates for a position as Assistant in Research to support the Materials Characterization Facility (MCF). The individual must have a Bachelor degree from an accredited institution in an appropriate area of study related to surface science and materials characterization, and have at least three years of experience in materials characterization or related areas. The person will be responsible for establishing and maintaining laboratory infrastructure, laboratory maintenance, equipment installation, operation and maintenance, and be an investigator in contracted research. The person must be able to conduct independent research and development in materials characterization with an emphasis in electron beam technology and have the ability to instruct students in the use and interpretation of data. The individual must have extensive knowledge and experience in the operation, maintenance and interpretation of materials characterization equipment. The salary range starts at $35,000 and is negotiable dependent on level and experience of the individual. The application opening date is December 18, 1998 and the closing date is December 31, 1998. Applicants should send a vitae and references to Dr. Lucille Giannuzzi, Director, Materials Characterization Facility, AMPAC Administrative Offices, 12424 Research Parkway, Suite 408, Orlando, FL 32826. UCF is an equal opportunity/affirmative action employer. As an agency of the State of Florida, all application materials and selection procedures are available for public review.
************************************************************************** Lucille A. Giannuzzi, Ph.D. phone: 407 823-4770 University of Central Florida fax: 407 823-0208 Materials Science Program email: lag-at-pegasus.cc.ucf.edu Dept. of Mechanical and Aerospace Eng. Orlando, FL 32816-2450 **************************************************************************
Scott asks ... } } ... } } I am evaluating FEG semi-in-lens SEMs for possible purchase. I am } interested in BSE imaging at low acc. voltages ( {3kV). ...
I am unfamiliar with the geometry for "semi-in-lens" vs a BSE detector which would need line-of-sight to the interaction spot on the sample. How does this work???
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Hello all We have recently had a number of users who have come from different places in the world, and use different conventions for whether sections/formar go on the shiny side of the grid or the dull side.
This lab uses the dull side, and has considered that the convention. If it ain't broke don't fix it.
As long as the user is consistent with his/her own grids, there is no problem, but when looking at collegues grids from another lab, it can lead to a few annoyances.
We have been making our own formvar or pioloform-coated grids and putting the formvar or pioloform on the dull side. One of my users has bought made-up-grids where the coating was on the shiny side. At least two other labs here use the shiny side for sections.
Is there a difference to the sides of the grid that makes one side better than the other? Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
We have recently had a number of users who have come from different places in the world, and use different conventions for whether sections/formar go on the shiny side of the grid or the dull side.
RESPONSE:
This is a good basic question - one that many microscopists have asked when coating grids with various plastic substrates. This question has even been studied in a scientific manner over the years and the conclusion has generally been that the dull side appears to hold on to the film more tenaceously than the shiny side. Now, I must admit that I started out using the shiny side and resisted going dull for many years (in reference to grids not sense of humor). I finally did a test and found that there was less drift and better film adhesion on the dull side.
Now a caveat: examine carefully the two sides of the grids and check if there are noticeable burrs or curvature to the grids (especially the latter). You should place the plastic film so that it is in maximal contact with the grid. So, if the grid is slightly bent, make sure the film goes on the convex side. Reason: on the concave side the film is suspended over the concavity and you will get a lot of drift. I have noticed that some slot grids have such a concavity and it may require you to place the film on the "shiny" side in this case.
So, generally speaking, place the film on the dull side of the grid unless the grid is dished or bent. In order to further facilitate adhesion of the film, some people place a drop of diluted plastic (like Formvar at 0.02%) on top of grids while on a filter paper. When dried, the thin plastic film has an affinity for the substrate or film that you finally place onto the grid for supporting sections, etc.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
{bold} {italic} Physics Today {/italic} {/bold} (Nov. 1998, p. 70):
"... this book is a unique, cutting-edge text on smart materials ... it is recommended as an adjunct to device design books used for engineers as well as scientists during the development of smart devices and structures."
{bold} {italic} {bigger} Science {/bigger} {/italic} {/bold} (Vol. 281, 10 July 1998, p. 181):
"The authors consider the atomic scale crystal structure and chemistry of oxides with physical and chemical properties that are sensitive to changes in the environment such as temperature, pressure, electric or magnetic fields, pH, and optical wavelength. They explain relationships among different structures and explore approaches to characterizing and synthesizing these important components for electronic devices"
{bold} {italic} COMMENTS FROM EXPERTS IN THE FIELD:
{/italic} {/bold}
"Both new in concept and timely in publication....Bring together, for the first time, the fundamental of atomic scale crystal structure and chemistry... A cutting-edge text at the forefront of the modern materials revolution"
- Professor David B. Williams, Lehigh University, USA
"Unique...focuses specifically on the intrinsic connections among several crystal structure systems and their evolution behavior...Fills a gap left in the field ... This book will be a basic reference in the domain of oxides which are to be the basis of functional and smart materials"
- Professor C. Boulesteix, Universite Aix-Marseille, France.
"In materials science the spotlight is on functional and smart materials, since they are important components for electronic devices. The textbook by Wang and Kang summarizes all types of known functional materials and describes the structure evolution problems. A large section of the book is devoted to structural characterization focusing on transmission electron microscopy, the main field of expertise of the author. The book is extremely valuable for materials scientists working on functional oxide materials, studying the structure, structure evolution and defects. It may serve also as an interesting textbook for teaching since it gives a good overview of this field which is of increasing importance. The clarity of its writing style should make it ideally suited for graduate students."
- Professor Manfred Ruhle, Institte of Werkstoffwissenschaft, Germany=20
Contents
Part I
STRUCTURE AND STRUCTURAL EVOLUTION=20
1. Structure, Bonding and Properties
1.1 Crystal structure
1.2 Structure and chemical composition
1.2.1 Stoichiometric phases
1.2.2 Non-stoichiometric phases
1.3 Coordination number and coordination polyhedron
1.4 Isotypism and polymorphism
1.5 Structure and chemical bonding
1.5.1 Bonding and ion radius
1.5.2 Lattice energy of an ionic compound
1.5.3 Geometric consideration of a structure
1.5.4 The rules of Pauling and Baur
1.5.5 Covalent bonding
1.6 Ligand field theory
1.6.1 Octahedral coordination
1.6.2 Tetrahedral coordination
1.6.3 Square coordination
1.7 Ligand field stabilization energy
1.8 Coordination polyhedra of transition metals
1.9 Molecular orbital theory of bonding
1.9.1 Molecular orbitals
1.9.2 Hybridization
1.10 Band theory
1.10.1 The Peierls distortion
1.10.2 Two- and three-dimensional bonds
1.11 Mixed valent compounds and functional materials
1.12 Structure transformation and stability
1.12.1 Phase diagram
1.12.2 Thermodynamic stability
1.13 Properties of materials
1.13.1 Mechanical Property
1.13.2 Magnetic property
1.13.3 Piezoelectric property
1.13.4 Ferroelectric property
1.13.5 Optical property
1.13.6 Electric property
1.14 Structure and property
1.15 Functional materials
1.15.1 Characteristics of functional materials
1.15.2 Structural evolution and functionality
1.16 Summary
2. Sodium Chloride and Rutile Related Structure Systems
2.1 Rock salt structure
2.2 Non-stoichiometric compounds with sodium chloride structure
2.3 Rutile structure and its family
2.4 Characteristics of rutile structures
2.5 Evolution of rutile-type structures
2.6 Non-stoichiometry and crystallographic shear planes
2.7 Summary
3. Perovskite and Related Structure Systems=20
3.1 Characteristics of ABO3 type perovskite structure
3.1.1 Vertex-sharing of oxygen octahedra
3.1.2 Unit cell by taking A cation as the origin
3.1.3 Oxygen cubic close-packing
3.1.4 Anion close-packing and formation of tetrahedron and octahedron
3.2 Possible types of anion-deficient perovskite structures
3.2.1 The 14 fundamental structure units
3.2.2 Constructing the family of perovskite related structures
3.3 The tolerance factor
3.4 Functional materials with perovskite-like structures
3.4.1 Ferroelectricity and ferroelectric compounds
3.4.2 Ferromagnetism and ferromagnetic compounds
3.4.3 Insulator to conductor transition
3.4.4 Conductive perovskites
3.4.4.1 Valence disproportionationality
3.4.4.2 Dimensionality
3.4.4.3 Building the structures of high temperature superconductors using perovskite structure units
3.4.5 Magnetostrictive, electrostrictive and piezoelectric actuator materials
3.4.6 Optically switchable compounds
3.5 Doping and oxygen vacancies
3.6 Giant magnetoresistance (GMR) and colossal magnetoresistance (CMR)
3.7 Oxygen migration and ionic conductivity of perovskites=20
3.8 Anion deficiency induced perovskite to brownmillerite structural evolution=20
3.9 Ordered structural evolution introduced by cation substitution
3.10 Sodium chloride, rutile and perovskite structures
3.10.1 Linkage and comparison
3.10.2 Constructing new materials by tailoring
3.11 Summary
4. Fluorite-Type and Related Structure Systems=20
4.1. Basic fluorite structure
4.2 Fluorite structure with anion deficiency
4.2.1 Oxygen migration in fluorite structure
4.2.2 Modules of fluorite structure with oxygen deficiency
4.2.3 Pyrochlores and C-type rare earth sesquioxide structures
4.3 Characteristics of fluorite and fluorite-related structures
4.3.1 Thermodynamic property
4.3.2 Surface character of rare earth oxides
4.3.3 Disproportionation of rare earth high oxides
4.3.4 Switchable chemical reaction as an oxygen pump
4.4 Structural and compositional principles of rare earth homologous higher oxides
4.4.1 Compositional principle of the homologoues phases
4.4.2 The modular juxtaposition rules
4.4.3 Building supercell structure using modules
4.5 Applications of the juxtaposition rules to known structures
4.5.1 R7O12 phase with n =3D 7 and m =3D 1
4.5.2 R9O16 phase with n =3D 9 and m =3D 1
4.5.3 R11O20 phase with n =3D 11 and m =3D 1
4.5.4 R40O72 phase with n =3D 40 and m =3D 4
4.5.5 R24O44 phase with n =3D 24 and m =3D 2
4.6 Predicting undetermined structures using the proposed modules
4.6.1 b-polymorph with m =3D 4
4.6.2 Undetermined structure with n =3D 19
4.6.3 Undetermined structure with n =3D 16
4.6.4 Undetermined structure with n =3D 62 and m =3D 6
4.6.5 Non-stoichiometric a-phase
4.7 Ternary mixed rare earth oxides
4.7.1 Rare earth mixed ternary oxides and oxygen storage
4.7.2 Cation coordination number and arrangements of modules
4.8 Perovskite, fluorite structures and spinel structures
4.8.1 Structure comparison
4.8.2 Superexchange interaction and magnetism
4.9 Summary
5. From Structural Units to Materials Engineering via Soft-Chemistry
5.1 Principle of soft chemistry
5.2 Sol-gel process
5.3 Colloidal route for preparation of monodispersive spherical particles
5.4 Intercalation and pillaring processes
5.5 Self-assembled nanocrystal engineered superlattice thin films
5.5.1 Passivated metal nanocrystals
5.5.2 Passivated semiconductors nanocrystals
5.5.3 Passivated magnetic nanocrystals
5.5.4 Magnetic Co particles
5.5.5 Magnetic iron oxides
5.6 Preparation of nanoparticles by aerosol technique
5.7 Summary
Part II
STRUCTURE CHARACTERIZATIONS
6. Electron Crystallography for Structure Analysis=20
6.1 Electron diffraction in structure analysis
6.1.1 Single scattering theory
6.1.2 Reciprocal space
6.1.3 Bragg's law and Ewald sphere
6.1.4 Indexing electron diffraction patterns
6.1.5 Diffraction from twinned crystals
6.2 Diffraction contrast and defect analysis
6.2.1 Defects and dislocations in materials
6.2.2 Diffraction contrast
6.2.3 Two-beam condition for imaging defects and dislocations
6.2.4 Determination of Burgers vector
6.2.5 Weak beam imaging
6.3 Atomic-resolution structure imaging and structure analysis
6.3.1 Phase contrast
6.3.2 Abbe's imaging theory
6.3.3 Aberration and information transfer in TEM
6.3.4 Contrast transfer function and image resolution
6.3.5 Envelope function and information transfer
6.3.6 Source coherence in lattice imaging
6.3.7 Projected charge density approximation
6.3.8 Multislice theory for transmission electron imaging
6.3.9 Image simulation and structure determination
6.3.10 Image calculation of imperfect crystal and interface
6.3.11 Energy-filtered electron lattice imaging
6.3.12 Limitation of HRTEM
6.4 Electron holography
6.4.1 Principle of off-axis holography in TEM
6.4.2 Improvement of image resolution
6.4.3 Imaging electrostatic field and charge distribution
6.4.4 Imaging spontaneous polarization at domain boundaries in ferroelectrics
6.4.5 Imaging magnetic domains and flux lines
6.5 Convergent beam electron microdiffraction
6.5.1 Symmetry analysis
6.5.2 Measurement of lattice parameters
6.5.3 Bloch wave theory and quantitative CBED
6.5.4 Solid state bonding and charge redistribution
6.5.5 Determination of Burgers vector
6.5.6 Measurement of specimen thickness
6.6 Summary
7. Structure analysis of functional materials=20
7.1 Interface and grain boundary analysis
7.1.1 Kikuchi pattern and grain boundary analysis
7.1.2 General description of a grain boundary
7.1.3 The O-lattice theory
7.1.4 Coincidence-site lattice theory
7.2 Modulation and domain structure
7.2.1 Structural modulation
7.2.2 Domains formed by anisotropic crystal structure
7.2.3 Boundaries of structure domains
7.3 Superstructure and long-range ordering
7.3.1 3-D superstructure analysis by a double-pattern technique
7.3.2 3-D superstructure analysis by a single-pattern technique
7.3.4 Long-range ordering of cation substitutions
7.4 Oxygen vacancies and short-range ordering
7.4.1 Kinematical diffraction theory of diffuse scattering
7.4.2 Geometrical description of diffuse scattering
7.4.3 Calculation of short-range ordering parameter
7.4.4 HRTEM study of short-range order
7.5 Effects of substrate on thin film growth
7.5.1 Lattice mismatch and interface dislocations
7.5.2 Nucleation and growth of defects from substrate surfaces
7.5.3 Linkage of domain boundaries with interface dislocations
7.5.4 Linkage of interface dislocations with planar defects
7.6 In-situ observation of structure evolution
7.6.1 Temperature driven structure transformation
7.6.2 Electric field driven structure transformation
7.6.3 Magnetic moment of the specimen
7.7 Failure analysis of devices
7.8 Imaging surfaces of oxides
7.9 Summary
8. Chemical and Valence Structure Analysis of Functional Materials
8.1 Inelastic excitation processes in electron scattering
8.2 Energy dispersive x-ray microanalysis (EDS)
8.2.1 Composition analysis
8.2.2 Atom location by channeling enhanced microanalysis (ALCHEMI)
8.3 Valence excitation EELS
8.3.1 Classical electron energy-loss theory
8.3.2 Surface plasmon excitation
8.3.3 Measurement of dielectric function
8.4 Atomic inner shell excitation in EELS
8.4.1 Composition analysis
8.4.2 Near edge fine structure and bonding in crystals
8.5 Quantitative determination of valences in a mixed valent=20 compound
8.5.1 White lines of transition metals
8.5.2 The occupation number of the d-band electrons
8.5.3 White line intensity and intrinsic magnetic moment
8.5.4 Double derivative spectrum for calculation of white line=20 intensity
8.6 Nano-probe analysis of interfaces and grain boundaries
8.7 Chemical sensitive imaging in STEM
8.8 Energy filtered electron imaging in TEM
8.8.1 Composition-sensitive imaging using valence-loss electrons
8.8.2 Composition-sensitive imaging using inner-shell ionization edge electrons
8.8.3 Mapping of bonding and valence state
8.9 Phonon scattering and chemical sensitive imaging
8.9.1 'Z-contrast' imaging in STEM
8.9.2 High-angle dark-field conical scan imaging in TEM
8.10 Conjunction use of various techniques for structure refinement of La8Sr8Co16O36 - an example
8.11 Summary
=09
APPENDIXES
A: Physical constants, electron wavelengths and wave numbers
B1: Crystallographic structure systems
B2: FORTRAN program for calculating crystallographic data
C: Electron diffraction patterns for several types of crystal=20 structures
D: FORTRAN program for calculating single valence-loss EELS spectra in TEM
Introduction
Smart systems and smart materials
Smart structures are a new emerging materials system which combines contemporary materials science with information science. The smart=20 system is composed of sensing, processing, actuating, feedback, self-diagnosing and self-recovering subsystems. The system uses the functional properties of advanced materials to achieve high performances with capabilities of recognition, discrimination, and adjustification in response to a change of its environment. Each component of this system must have functionality, and the entire system is integrated to perform a self-controlled smart action, similar to a living creature who can "think", make judgment and take actions. A smart system can be considered as a design philosophy that emphasizes predictivity, adaptivity and repetivity.=20
A smart system/structure is defined to be a non-biological physical structure having the following attributes: (1) a definite purpose; (ii) means and imperative to achieve that purpose; and (iii) a biological pattern of functioning (Spillman et al., 1996). Smart materials are a subset of the smart system, i.e. smart structures at the microscopic or mesoscopic scales. Smart system is a non-biological structure which means that the system functions as a biological system rather than the pattern of functioning of a Turning machine. A smart material is a physical structure having (i) a definite purpose, (ii) means of imperative to achieve that purpose, and (iii) the pattern of functioning of a universal computer or Turning machine. Such a material will generally include at least one structural element, some means of sensing the environment and/or its own state, and some type of processing and adaptive control algorithm.=20
Science and technology in the 21st century will rely heavily on the development of new materials that are expected to respond to the environmental changes and manifest their own functions according to the optimum conditions. The development of smart materials will undoubtedly be an essential task in many fields of science and technology such as information science, microelectronics, computer science, medical treatment, life science, energy, transportation, safety engineering and military technologies. Materials development in the future, therefore, should be directed toward creation of hyperfunctional materials which surpass even biological organ in some aspects. The current materials research is to develop various pathways that will lead the modern technology toward the smart system.=20
Functional materials
Functional materials are distinctly different from structural materials, and their physical and chemical properties are sensitive to a change in the environment such as temperature, pressure, electric field, magnetic field, optical wavelength, adsorbed gas molecules and the pH value. The functional materials utilize the native properties and functions of their own to achieve an intelligent action. Functional materials cover a broader range of materlas than the smart materials illustrated above. Besides the materials belogning to the smart structure, any materials that have functionality are attributed to functional materials, such as the ferroelectricic BaTiO3, the magnetic field sensor of La1-xCaxMnO3, surface acoustic wave sensor of LiNbO3, liquid petroleum gas sensor of Pd-dopped SnO2, semiconductor light detectors (CdS, CdTe), high temperature piezoelectric Ta2O5, fast-ion conductor Y2(SnyTi1-y)2O7 (pyrochlore structure), the electric voltage induced reversible colouring of WO3, and high temperature superconductors etc. Functional materials cover a wide range of organic and inorganic materials. This book focuses only on oxide functional materials.
In recent years, techniques for epitaxial crystal growth have made it possible to grow oxides and metal thin films on silicon substrates, and this is the first step to integrate functional materials with the logic system. Preparations of complex oxides with functionality are a key challenge for materials development. Searching new routes to prepare materials and understanding the relationship between the structures and the properties are equally important. A key requirement in preparations of materials is to control the structural and compositional evolution for achieving superior properties. "Soft chemistry" has shown a great success in fabricating functional and nanophase materials. Nanocrystal engineered materials are a new trend of materials research, aiming to improve the performances of materials by several orders of magnitudes.=20
Mixed valences and functionality
Crystal structure usually refers to two aspects of information, one is the symmetry and distribution of atoms in the unit cell and the other is the bonding between atoms. Thermordynalically, the enthalpy of the system is determined by the bonding between atoms, while the entropy is determined by the atomic lattice configurations of the crystal. Thermodynamic rules select the possible stable phases, and the phase stability is strongly affected by bonding. A single element has a certain electron configuration. When several different elements form a molecule, the electronic structure of this cluster is very different from any of its original elemental configuration because of the transfer and/or sharing of valence electrons among atoms. In general, only the valence electrons are most critical to bonding, the distribution and motion of valence electrons are usually described by the molecular orbitals. These valence states and molecular orbits are responsible for the functional properties of the molecule. The ligand field theory is designed to describe the molecular structure of an atom cluster.=20
When different elements are combined to form a crystalline solid in which the atoms or atom groups (or molecules) are bonded together to form a three-dimensional (3-D) structure with specified symmetries, the properties of the solid would depend on both the electronic structure of the atoms or atom groups and their spatial distribution. The molecular orbital theory and band structure theory are usually applied to elucidate the relationship between the structure and the properties. Based on the electron band structure, inorganic materials can be classified as conductors, semiconductors and insulators. If a change is made in the crystal structure so that the band gap is reduced or eliminated, a transition from an insulator to a conductor is possible. Modifying a crystal structure can be performed by changing either the spatial distribution of atoms (such as bonding angles, bonding lengths and symmetry of atom arrangement) or chemical composition (such as from stoichiometry to nonstoichiometry). All these changes are referred to as structural evolution, which is closely related to the properties of the materials.=20
Many functional properties of inorganic materials are determined by the elements with mixed valences in the structure unit, by which we mean that an element has two or more different valences while forming a compound. Valence mixture refers to a case in which several elements have different valences but each one only has a single valence. In the periodic table of fundamental elements, 40 elements can form mixed valent compounds, transition d-block elements and lanthanide (Eu, Yb, Ce, Pr, Tb etc.) are typical examples. Modern inorganic chemistry has shown that the oxidation state of any element can be modified under special conditions. Many oxide functional materials contain elements with mixed valences. This is a typical difference between the functional materials and the structural materials. This book is about the structural evolution of compounds containing mixed valent elements, such as transition and rare earth elements. =20
The concept of mixed valent chemistry offers a pathway to design and synthesize new compounds with unique optical, electric, or magnetic properties. Research in functional materials in its broad sense always depends on the conception and synthesis of interesting novel mixed-valent compounds. The discovery of high temperature superconductor compounds is a fascinating successful example of the mixed valence chemistry. We believe that exploring the possible structures of mixed valent compounds and their evolution behaviors may lead to many pathways to synthesis new functional materials.=20
The scope of the book
Searching for new functional materials is a challenge for the development of smart systems. To guide this searching, a clear understanding about the relationship between the physical properties and the atomic-scale structure of the materials is desperately needed. This book is about the intrinsic connections among several crystal structure systems commonly used in functional materials and their evolution behaviors. The book is not intended as a source for listing all the existing functional materials, instead its goal is to reveal the principles for engineering and controlling functional materials from the fundamental structure units. The functional materials are described from the mixed valences and stoichiometry points of view to understand the structural evolution and transformation of different materials systems. The mixed valent compounds are elucidated as the fundamentals for performing unique functionality.=20
We have written this book with a strong conviction that functional materials system is a future direction of the multidisciplinary research involving physics, chemistry, materials science, electrical engineering and biological science, with an emphasis on molecular and unit cell designs. There are numerous books describing the properties, preparations, electronic structures and crystal structures of transition and rare earth metals and their oxides. To expert in the field, this book is written by addressing the issues that have not been described systematically in existing books. Our aim is to explain the intrinsic connections among different structures. The goal is to explore new routes for synthesizing functional materials from the fundamental structure building blocks (or modules). The book aims to illustrate not only the role played by crystal structure in property control of functional materials, but also the structure determination using advanced transmission electron microscopy and spectroscopy techniques. The latter also has critical importance for device failure analysis in modern industry.
Accordingly, the book is written into two parts. Part I concentrates on the structure and structural evolution of oxides functional materials belonging to NaCl-, rutile-, perovskite and CaF2-type structures and the related. Although our analysis is focused on the structure systems outlined above, spinel and corundum structures are also briefly described in these chapters except the wurtzite structure (BeO and ZnO) because of its limited evolution characteristics. Part II is on crystallographic and chemical structure characterizations of oxide functional materials, which are needed to understand the experimental approaches for exploring the structure evolution. Both parts are written to ensure the coherency of the whole book.
Part I is composed of five chapters. In Chapter 1, fundamental concepts are introduced on crystallographic structures, chemical structures, bonding, molecular orbital and ligand field, mixed valences, materials properties and the fundamental characteristics of functional materials. This chapter is designed as the preliminary preparation for the discussions to be outlined in future chapters. The characteristics of functional materials are given for distinguish them from structural materials. Chapter 2 starts with the simplest oxides with NaCl structure for illustrating the stacking of cations and anions in constructing the unit cell. Then, rutile related structures are introduced with an emphasis on their structural evolution. Chapters 3 is on perovskite and related structures. The characteristics of the ABO3 perovskites are thoroughly analyzed to reveal the intrinsic connections among the A, B and O elements and the roles played by the octahedron in constructing the unit cells. The alternative stacking of the close-packed (AO3)4- and B layers is analyzed in detail, and a total of 14 fundamental structure units are extracted with introducing of anion deficiency, which are the building blocks for constructing the unit cells of complex functional materials. The tailoring of cation octahedra can give the structure of a variety of compounds. The typical properties of perovskites are also described and their relations with the crystal structure are elucidated. This chapter is the basis for understanding the mechanisms that control cation substitution, creation of oxygen vacancies and mixed valences in the perovskite family.
Similarly, the fluorite related structures are analyzed in Chapter 4, and a total of 12 fluorite modules (or units) with anion deficiencies are proposed, which together with the perfect fluorite are the 13 basic building blocks for constructing the structures of rare earth homologoues higher oxides. Following the structural and compositional principles outlined by Kang and Eyring (1995, 1996), geometrical assembling of these modules can reproduce not only the known crystal structures of homologoues phases RnO2n-2m but, more importantly, predict the structures of some phases whose structures are unknown. This chapter proves that a complex superstructure can be disassembled into some fundamental modules, which can be derived at the first place from the basic fluorite structure. Therefore, by revealing the intrinsic connection among the same class of phases one may be able to predict and design new structures. The surface property and oxygen migration can be better understood from the view of module combination. At the end of Chapter 4, the structures of perovskite, fluorite and spinel are compared with each other and a brief introduction about spinel is given.=20
Chapter 5 focuses on the introduction of soft-chemistry (sol-gel, pillaring and grafting, intercalation and deintercalation) for synthesizing and designing new materials based on the fundamental structure modules. The chapter aims to using the understanding about structural evolution described in previous chapters to develop new materials systems that are expected to have functionality. Preparation of nanophase materials is also described with an emphasis on self-assembled nanocrystal superlattices (or arrays). This is a new trend of materials research on nanocrystal engineered (or patterned) materials.
Part II is composed of Chapters 6-8. Chapter 6 aims to introduce the fundamentals of structure analysis using transmission electron microscopy and associated techniques, including electron diffraction, diffraction contrast for defect analysis, atomic resolution lattice imaging for interface studies, electron holography for studying ferroelectric and ferromagnetic materials, and convergent beam electron diffraction technique for mapping charge redistribution and bonding in crystalline materials. While introducing these fundamental techniques, our focus is on the applications of these techniques for the analysis of functional materials. This chapter also serves as an educational chapter to the readers who have different background in solid state chemistry, materials processing and structure analysis, for correctly using the available tools for solving the problems in oxide functional materials. The applications of the techniques introduced in Chapter 6 are described in Chapter 7 case-by-case to cover the structural characteristics of oxide functional materials, including grain boundaries, interfaces, domains, structure transformations and surfaces. Analysis of long-range superstructures and short-range order of point defects is the emphasis of this chapter because they are closely related to the structural evolution described in Chapters 2-4. Numerous examples are shown to illustrate the techniques for solving the structure problems belonging to functional materials.
Chapter 8 focuses on the chemical and bonding analysis of mixed valent oxides at a high spatial resolution using the energy dispersive x-ray spectroscopy (EDS) and electron energy-loss spectroscopy (EELS). The near edge fine structure observed in EELS is a sensitive technique for detecting the valence band structure from a small region, and it also allows the analysis of interface electronic structures. The observed white lines in EELS can be used to determine the occupations of the 3d and 4d orbitals (e.g., valence states) of transition and rare earth elements. This information together with the imaging and diffraction data from HRTEM can be applied to determine the ordered structure induced by ordered anion vacancies. Finally, it is emphasized that the structure information provided by imaging and diffraction techniques (Chapters 6 and 7) must be integrated with the chemical information provided by EDS and EELS. The structure refinement of an anion deficient phase La8Sr8Co16O36 is demonstrated as an example. A combination of TEM results with x-ray diffraction or neutron diffraction data and theoretical modeling as well is likely to give a unique and reliable solution to a material's problem. It is concluded that the structural evolution in a complex system is intrinsically dominated by the combinations of the fundamental structural modules. This is the focus of this book.
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Reply to: RE: grids dull or shiny?
Elaine Humphrey wrote: } Hello all } We have recently had a number of users who have come from different = places } in the world, and use different conventions for whether sections/formar = go } on the shiny side of the grid or the dull side. } } This lab uses the dull side, and has considered that the convention. If = it } ain't broke don't fix it. } } As long as the user is consistent with his/her own grids, there is no } problem, but when looking at collegues grids from another lab, it can = lead } to a few annoyances. } } We have been making our own formvar or pioloform-coated grids and putting } the formvar or pioloform on the dull side. One of my users has bought } made-up-grids where the coating was on the shiny side. At least two = other } labs here use the shiny side for sections. } } Is there a difference to the sides of the grid that makes one side better } than the other? } Elaine } } } Dr. Elaine Humphrey } Biosciences Electron Microscopy Facility } University of British Columbia } 6270 University Blvd, mail-stop Botany } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-unixg.ubc.ca } Reply;
Ter trivial answer to this question is that all experts are correct. = Therefore either side is good. However, there is one instance where I = have found this to be incorrect. Single slot grids have a polarity to = them. The dull side has rounded edges wherease the shiny side is = completely flat. Check it out. the immediate concequence is that if = formvar is applied to the dull side it will eventually sag into the hole = and stick to the support you store it on. On the shiny, flat side, this = is not a problem. This might be true for the mesh grids too. I can't say = more on this because I never looked. =
For the record, for routine use I usually put the film on the dull side = too and have never had a problem. I know a colleague who swears that using = the shiny side is best because it improves resolution! I do know that if = the film has been damaged in some way then buffers will react with the = metal. Hence the dogma of not using copper grids for immunocytochemistry. = =
My advice is to believe all the experts and do what ever you want. Even = better, become an expert and make everyone in your lab do it your way! } } } } RFC822 header } ----------------------------------- } } Received: from Sparc5.Microscopy.Com [206.69.208.10] by mailhouse.hei.= org } (SMTPD32-4.07) id A8EC245B023C; Fri, 04 Dec 1998 21:47:24 PST } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.= 11) = } id RAA29721 for dist-Microscopy; Fri, 4 Dec 1998 17:19:29 -0600 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id RAA29715 for = } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 4 Dec 1998 17:18:58 -= 0600 } Received: from mail.unixg.ubc.ca (mail.unixg.ubc.ca [137.82.27.14]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA29707 for = } {microscopy-at-sparc5.microscopy.com} ; Fri, 4 Dec 1998 17:18:44 -0600 } Received: from cbig-se.botany.ubc.ca ([137.82.136.14]) } by mail.unixg.ubc.ca with esmtp (Exim 1.92 #1) } for microscopy-at-Sparc5.Microscopy.Com } id 0zm4h8-0000tc-00; Fri, 4 Dec 1998 15:31:30 -0800 } X-Sender: ech-at-pop.unixg.ubc.ca } Message-Id: {v0300780be8f1d9725816-at-[137.82.136.14]} } Mime-Version: 1.0 } Content-Type: text/plain; charset=3D"us-ascii" } To: microscopy-at-Sparc5.Microscopy.Com } From: Elaine Humphrey {ech-at-unixg.ubc.ca} } Subject: grids dull or shiny? } Date: Fri, 4 Dec 1998 15:31:30 -0800 } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-UIDL: 907881843 } Status: U } =
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org
by arl-img-12.compuserve.com (8.8.6/8.8.6/2.16) id HAA12219; Sat, 5 Dec 1998 07:04:07 -0500 (EST)
Hi Guys
Maybe I can help?
Semi in lens imaging, or dual detector imaging, relies upon the SE being attracted to an detector which is situated above the final lens pole piec= e. Initially commercial instruments with two ET detectors used the field fr= om the final lens to "pull" SE into the upper detector. ISI were first with=
their SS Series and then Hitachi with the S570 took this route. In these=
instruments they allowed the specimen to be anywhere between 100% out of lens through to what we would consider to be a WD of -5mm. That is 5mm u= p inside the pole piece! Problems arose if you were using magnetic materials, hence the development of a twin detector system by Hitachi whi= ch used a field coil to drag electrons up to the detector and a pole piece design which retained the lens field within the pole piece. =
The twin detector system gives the microscopist the best of all worlds.
1. The upper detector provides an opportunity to sift out the BSE an= d obtain a pretty pure SE image. The high lens strength at very short WD (~3mm) enables the instruments to reach very high resolution levels. The down side of this is, as SE are effected by charge, this mode is prone to=
charge problems.
2. The lower detector offers the type of contrast which we all see i= n our conventional single detector SEM, SE+BSE. The up side is that the BS= E contribution to this image results in far fewer charge problems.
3. Add a BSE detector to such a system, and you can move in any direction "pure" SE or SE + BSE or pure BSE. Drop the kV and the BSE becomes even more interesting as the volumes of material involved almost mimic SE volumes.
All of you interested in image formation should be very jealous of someon= e who will be able to get his hands on an instrument like this; 100% scienc= e!
When testing instruments of this type be very careful to select specimens=
that do look different at different kV. Gold on carbon is in my mind a very poor performance standard for this type of instrument. It is often unable to pull out the performance differences between 5 and 15kV. This does not demonstrate a fantastic instrument but a poor test specimen.
You lucky man!
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
The people I would contact in the FEG + BSE area are Geoff Richards and Iolo ap Gwynn. They are very active in the biological side of BSE imagin= g.
=46rom by own experiences you have no substitute for beam current, you ne= ed the ability to drive the gun at least up to 20uA, you will find some manufactures even have 40uA if you know where to get at it!
Try Iolo ap Gwynn on iag-at-aber.ac.uk
Steve Chapman Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Well after a great Conference and some foul weather what better than a go= od game of rugby?
We have to say that a really GREAT team were beaten on the day by a very good team. Sorry you did not break the record (although your TV commentator would have been moaning about the ref as he always seems to d= o) but you ALL have to admit that we beat you fair and square?
Dr. Elaine Humphrey wrote: ============================================ We have recently had a number of users who have come from different places in the world, and use different conventions for whether sections/formar go on the shiny side of the grid or the dull side.
This lab uses the dull side, and has considered that the convention. If it ain't broke don't fix it.
As long as the user is consistent with his/her own grids, there is no problem, but when looking at collegues grids from another lab, it can lead to a few annoyances.
We have been making our own formvar or pioloform-coated grids and putting the formvar or pioloform on the dull side. One of my users has bought made- up-grids where the coating was on the shiny side. At least two other labs here use the shiny side for sections.
Is there a difference to the sides of the grid that makes one side better than the other? ===================================================== This is certainly the most often asked-for question about TEM grids, at least for our firm, from customers writing or calling in for "advice" .
After being involved for more than thirty years with this question, I have come to the following conclusions. I present them here for whatever peer review might be forthcoming:
a) It seems to be about 50/50 (non-scientifically determined) on a worldwide basis as to whether users prefer dull or shinny.
b) While the "dull" side has sharper "protrusions" and in theory at least should initiate tears and cracks more readily than the shinny side, I am not sure we ourselves have ever found evidence that is the case. Those expressing preferences for the dull side tend to claim better adhesion because of the larger surface area available for adhesion. So we have ourselves given up on using our own good common sense to rationalize which side is "better". We think that one can, in general, get comparable results from either side.
c) However, not all grids are manufactured using the same processes and the conclusions drawn from grids from one source might not necessarily be applied to grids from other sources. For example, anyone who has used different "brands" of grids knows that there is a difference between dull and shinny side discrimination. We have even found customers who have expressed a preference for grids showing the least dull/shinny difference so it is not necessarily even a case of good vs. bad.
d) We ourselves believe that the real point of the dull vs. shinny side is to quickly and easily discriminate between which side is "up" and which side if "down" since many samples are not easily seen with the naked eye. Some percentage of the population develop a level of color-blindness, and this is also true as we get older, and as that happens, there also seems to be a loss in ability to discriminate between dull vs. shinny. Hence, some grids are actually purchased on that basis alone, without regard to any other considerations. From that perspective it does not matter which side is being used, just so one is consistent and always using the same side for their specimens.
e) Paul Webster raises an interesting question relative to there being an asymmetry (he called it a polarity) to the slot grids. We ourselves have never looked at that aspect of the issue before. But if it is correct, then it would probably also apply to the use of aperture grids as well, since they are made under identical conditions. In general, it is the shinny side that is the one in contact with the "glass master" during the grid making, and the dull side is the one that "grows" out into the plating bath. In any case the point is well made and this will encourage us to see what could be done to reduce the asymmetry present in slot grids.
f) All of my comments and I presume those of the others are directed specifically for those grids that are electro deposited, that is, Cu, Ni, and Au and such comments would not be applicable to grids made by other methods (for example, chemical etching, to produce grids from Be or W, etc.)
Dr. Rik Brydson wrote: ===================================== Some information on a new listserver recently set up. I encourage you to join Please display in your institution.
Many thanks, ================================================ It is not clear to me how this new listserver would in some way contribute to the objectives as outlined, in ways not possible via the existing listserver sponsored by the Microscopy Society of America. Right now, if someone has a question, they need post it in only one place, if the "places" were fragmented, to get the same full coverage, one would have to place it in more than one place.
And for those who felt they wanted to subscribe to more than one listserver, they are going to be reading the same postings more than one time.
I would cast my vote that the system we have "ain't broken" and therefore we shouldn't try to fix it!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
But the higher currents needed might upset your samples, mightn't they?
rtch
} Jacob Bastacky wrote: } } } } We are considering looking for a Wavelength-Dispersive Spectrometer to } } improve our detection of sodium in biological samples, reasoning that the } } better energy resolution of the detector will result in narrower, taller } } peaks with the same background level, yielding improved peak-to-background } } ratios. } } This is correct. } } } } Our samples are frozen hydrated and dried biological tissues; we would like } } to be able to differentiate 50mMolar from 150mMolar sodium (0.25 and .75% } } by weight in hydrated samples). } } } This should be pretty straight-forward. } } } } Might we expect a significant improvement with WDS? } } } Yes.
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
The bars on the shiny side have sharper edges and smoother surfaces than the bars on the dull side.
Check this out with your SEM
The argument for using the dull side for the films is that there is less risk of tearing at the edge of the bar and perhaps better adhesion.
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
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One final thought on this:
Interference colors, which reveal the location of sections on the grid, = are more easily seen on a dull background. Interference colors also play = an important role in the evaluation of support film thickness during the = final, drying step in cryosectioning. There is a reason for choosing one = side after all.
Regards, =
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/apw.htm
by imo22.mx.aol.com (IMOv18.1) id 8BYIa17651 for {microscopy-at-sparc5.microscopy.com} ; Sun, 6 Dec 1998 17:56:54 -0500 (EST) Message-ID: {d2de1999.366b0bb6-at-aol.com}
Readers, Due to the considerable number of request, we have decided to publish the results of our recent microscopist salary survey. However, do to the tabulation, I regret that I do not know how to publish it on this medium. The full survey results will be in the December issue of Microscopy Today - which will hit the P.O. on 17 December. If you have provided your salary data and wish an earlier copy, send me your fax number and I will be pleased to fax you a copy. Don Grimes, Microscopy Today P.S. #1: A surprise (to me) in the survey! The average yearly salary of all 457 microscopists reported was $51,676. The average yearly salary of all 268 male microscopists was $55,641 - or up some 7.7 % from the total average. But, the average yearly salary of all 189 female microscopists was $46,053 - or down some 10.9 % from the total average. No conclusion suggested! Interesting? P.S. #2: My sincere wishes for a nifty set of holidays to all - particularly, Nestor, to you!
Elaine: Years ago I read a brief SEM study which looked at the attachment of films and sections and concluded that the dull side was slightly better. However, the grid bars edges are a little more rounded on the dull side and supporting films or sections would tend to go a bit more up and down with that rounding. I always used the dull side, but what matters is uniformity in one working group. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, December 05, 1998 9:32 AM, Elaine Humphrey [SMTP:ech-at-unixg.ubc.ca] wrote: } } ---------------------------------------------------------- } -------------. } } } Hello all } We have recently had a number of users who have come from } different places } in the world, and use different conventions for whether } sections/formar go } on the shiny side of the grid or the dull side. } } This lab uses the dull side, and has considered that the } convention. If it } ain't broke don't fix it. } } As long as the user is consistent with his/her own grids, } there is no } problem, but when looking at collegues grids from another } lab, it can lead } to a few annoyances. } } We have been making our own formvar or pioloform-coated } grids and putting } the formvar or pioloform on the dull side. One of my users } has bought } made-up-grids where the coating was on the shiny side. At } least two other } labs here use the shiny side for sections. } } Is there a difference to the sides of the grid that makes } one side better } than the other? } Elaine } } } Dr. Elaine Humphrey } Biosciences Electron Microscopy Facility } University of British Columbia } 6270 University Blvd, mail-stop Botany } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-unixg.ubc.ca } }
Yes, WDS will be better for sodium but the trade off you have to make is that you will probably have to put in more beam current to get sufficient signal. A windowless or an atmosphere thin window (100nm) should work okay. Depends what else is in the spectrum. I would try it on a known standard. I think you are dead in the water with a beryllium window, too many of the sodium x-ray photon would get absorbed.
Best Wishes for the Christmas Season
Look forward to seeing you next summer. Shirley and I will be on the West Coast.
Patrick
On Thu, 3 Dec 1998, Jacob Bastacky wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are considering looking for a Wavelength-Dispersive Spectrometer to } improve our detection of sodium in biological samples, reasoning that the } better energy resolution of the detector will result in narrower, taller } peaks with the same background level, yielding improved peak-to-background } ratios. } } Our samples are frozen hydrated and dried biological tissues; we would like } to be able to differentiate 50mMolar from 150mMolar sodium (0.25 and .75% } by weight in hydrated samples). } } Our present EDS detector has a semi-thin window and is doing better than } average for sodium and we would prefer not going to an ultrathin window or } windowless detector. } } Might we expect a significant improvement with WDS? } } Does anyone know of a spectrometer that might be donated? } } Your comments and suggestions will be appreciated. } } Thanks, } } Jacob } } Jacob Bastacky, M.D. } Room 116 Donner Laboratory } Lawrence Berkeley Laboratory } University of California } Berkeley, California 94720 } Telephones: 510.486.4606 office, 510.486.4605 lab, 510.845.8031 fax } email: sjbastacky-at-lbl.gov } } } }
Dear Listers, I am seeking your guidance on using EDS to quantitate the ion uptake in homogeneous polymers (cast at a uniform thickness). Any references would be appreciated as well. Rosemary
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I would like to hear from microscopists who have used FEG-SEM's and particularily the semi-in-lens models for biological applications. I would also appreciate info on recent papers where this instrumentation was critical to the biological research reported. Also any recommendations as to ideal samples for testing these microscopes for biological application? Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------
I am evaluating FEG semi-in-lens SEMs for possible purchase. I am interested in BSE imaging at low acc. voltages ( {3kV). I would like to get some feedback as to people's experiences with using BSE mode at low voltages. Specifically, what kind of detectors are being used and what kind of results? What is the lowest acc. voltage that you can reasonably use and still get good results. Thanks,
RFC822 header -----------------------------------
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by arl-img-4.compuserve.com (8.8.6/8.8.6/2.16) id JAA07014 for microscopy-at-sparc5.microscopy.com; Mon, 7 Dec 1998 09:38:06 -0500 (EST)
Dear Dorrance,
To start with, I should mention that I am an interested party since =
BUEHLER is a major supplier of polishing equipment, consumables, and technical support for the grinding/polishing industry and has been for over 60 years.
To my point: Although I agree with Gary Leichty's point about using diamond =
vs. silicon carbide papers, it is for different reasons. Silion Carbide = is a brittle abrasive with a high coefficient of friction compared to diamond. Due to=
this fact, and as a result of being fixed to a paper backing material, SiC tends to 'plow' material during grinding operation. It also tends to fracture, creating smaller particles that can embed in ductile phases. Since the SiC abrasive is rigidly fixed to the paper backing material, when it 'plows' into the metallic phase of your alloy, it eventually contacts a hard precipitate and plucks it out.
Diamond films also 'plow' ductile phases because the abrasive is fixed to= a
backing material. In this case, as opposed to SiC papers, the resin bind= er of the film is weaker and allows the diamond to pull free of the film and stay embedded = in the sample. =
Although proper lubrication will help, diamond films are not the answer i= n your case. =
A better suggestion might be to section a disk from the bulk using a thin=
diamond blade, making sure that the sample is near final thickness. Then=
use one or more sizes of diamond paste product on a porous pad for removi= ng
damage created by the blade. The waxy carrier of paste products along wi= th embedding of the diamond in the pad will tend to eliminate embedding in your sample.
Finally, using a low napped polishing cloth with a colloidal silica produ= ct (High pH) will produce a smooth, low deformation surface in which no embedding =
exists, and in which both the aluminum and precipitate phases are relatively coplanar.
Please contact me directly if you have additional questions.
Best regards,
Scott D. Holt BUEHLER, Ltd. PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-4546 ****** http://www.buehlerltd.com ******
} Dear All, } } I've been batting my head against my Fischione Jet Polisher trying to figure } this one out! Please help if you can. My original material is full of=
} lovely voids and pits, Si particles are around 10 microns, and at a } thickness of 150 to 200 microns I have experienced some holes that go a= ll } the way through the material. If I try initial thinning of the sample = on } less than 500 grit SiC paper I get even more particle pullout...Any ideas? } } Thanks in advance for any help. } } Dorrance
So far the listserver has been a good source of infomation. I think we would all lose if we splinter into smaller listservers. If anything I would consider using better (i.e. clearer) Subject Identifiers. For example, one could use headers such as Bio (for Biological), Mat (for Materials ) and Gen. (for general ) subjects. This would make it easier for a reader to decide if he/she wants to open a given posting. I believe something like this was proposed by others in the past.
We also get much useful information from vendors (I am not a vendor) and if we were to split this would pose increased problems for both vendors as well as us microscopists in terms of the time spent searching and posting messages.
In short, I think we would lose more than what we would gain.
Jordi Marti
---------- } From: Garber, Charles A. To: MICROSCOPY BB -----------------------------------------------------------------------.
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Dr. Rik Brydson wrote: ===================================== Some information on a new listserver recently set up. I encourage you to join Please display in your institution.
Many thanks, ================================================ It is not clear to me how this new listserver would in some way contribute to the objectives as outlined, in ways not possible via the existing listserver sponsored by the Microscopy Society of America. Right now, if someone has a question, they need post it in only one place, if the "places" were fragmented, to get the same full coverage, one would have to place it in more than one place.
And for those who felt they wanted to subscribe to more than one listserver, they are going to be reading the same postings more than one time.
I would cast my vote that the system we have "ain't broken" and therefore we shouldn't try to fix it!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Dear Colleagues Being aware of your interest in 3-D reconstruction and visualization of biological object structures, I wonder if you deal with the problem of 3-D epithelial sheets topology reconstruction? If it is so can you send me your papers on this subject? And do you know anybody who work in this field? My own interest is the 3-D histoarchitecture of epithelial sheets in normal development and pathology. In this connect I am addressing you with the following message. As you know, all current attempts to reconstruct a 3-D structure of biological tissues using a serial sections encounter with a typical difficulty. It is that tissue deformation and cell proliferation make a cells shape variable, diverse their adjacency and give a strongly "noised" pictures. So the results obtained give no way to understanding tissue topology and do not permit to deduce the law of 3-D tissue organization.
To overcome the difficulties we have developed the new approach to 3-D reconstruction of cell sheet structures. It is based on the pioneering concept of tissues modular structure and consists of derivation of topological and geometrical models of epithelial spatial organization and their experimental verification. Contrary to the usual tissue reconstruction on a set of serial sections this approach allows to use their minimum and to carry out a more exact restoration of 3-D epithelial structure with less money and time. The approach has enabled us to get the data priority on the laws of spatial organization of simple, pseudostratified and stratified epithelia, as well as to predict and find several earlier unknown topological variants of their histoarchitectonics. The approach has also allowed to describe such new properties of epithelial layers as translation symmetry and stoichiometry.
The results makes a basis for structural histology as a addition to modern structural biology. The approach allow to investigate the more complex topology variants of pseudostratified and stratified epithelia, to find a set of new informative tissue characteristics suitable for diagnostic and also to give the opportunity to predict the changes of tissues in normal development and their transformation in pathology, particularly in malignant growth.
If it is interesting for you, I am ready to consider the possibility of our cooperation.
Yours sincerely, Dr. G.A. Savostyanov. -- | E-mail: savost-at-ief.spb.su | Gennady A. Savostyanov | | Fax +7 (812) 5523012 |Sechenov Inst. of Evolutionary Physiology and | | office +7 (812) 5523090 | Biochemictry Russian Academy of Science, | | | home +7 (812) 5100052 |44 Thores av., 194223, St.Petersburg, Russia |
What is the purpose of your having to section? If it is to determine pigment distribution, have you tried confocal? As long as there is not titantium dioxide as a filler (it tends to act as a mirror), confocal may work, saving you lots of aggrevation.
If you have to section, talk to Wasi Ahmed at Buehler Instruments here in the States. He had sectioned everything from light bulbs to the ash on the end of a cigar! Try them at 847-295-6500.
} P.S. #1: A surprise (to me) in the survey! The average yearly salary of all } 457 microscopists reported was $51,676. The average yearly salary of all 268 } male microscopists was $55,641 - or up some 7.7 % from the total average. } But, the average yearly salary of all 189 female microscopists was $46,053 - } or down some 10.9 % from the total average. No conclusion suggested! } Interesting?
Not interesting, just sad. No matter how good we are or how hard we try, women still are considered second class citizens and our salaries reflect that. We're used to, but not happy about, being paid less almost 100% of the time.
Paula :-(
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Dear All. How can I calibrate the maginfication in a scanning transmission electron microscop for } 100.000x? Up to this mag one can use cross grating replica, but beyond? Thank you very much. -- Gerhard Frank UNIVERSITAET ERLANGEN-NUERNBERG Phone: +49 9131 85 28606 Institut fuer Werkstoffwissenschaften VII Fax: +49 9131 85 28602 Cauerstr. 6, D-91058 Erlangen, Germany frank-at-ww.uni.erlangen.de ***************************** * NEW Phone and Fax numbers!* *****************************
P.S. #1: A surprise (to me) in the survey! The average yearly salary of all 457 microscopists reported was $51,676. The average yearly salary of all 268 male microscopists was $55,641 - or up some 7.7 % from the total average. But, the average yearly salary of all 189 female microscopists was $46,053 - or down some 10.9 % from the total average. No conclusion suggested! Interesting?
Not interesting, just sad. No matter how good we are at what we do or how hard we try, women still are considered second class citizens and our salaries reflect that. We're used to, but not happy about, being paid less almost 100% of the time.
Paula :-(
p.s. everytime I try to reply to the MicroToday-at-aol.com-at-sparc5.microscopy.com I get a bad address and the message is thrown back, does anybody know why?
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
There's another option of course, there is a newsgroup titled alt.sci.microscopy. My ISP seems to have a problem w/ this one though, casue I lose all the threads in about a week.
It works basically the same was as this listserver, only you have the option of reading or not reading messages, rather than having all messages sent to you via email.
When i have a question or such, I'll usually post in both places. For example, I posted my questions regarding the TN5500 here and there. I recieved a few responses from there and many more from here. I am not sure, however, how many people know of the newsgroup.
{!doctype html public "-//w3c//dtd html 4.0 transitional//en"} {html} There's another option of course, there is a newsgroup titled alt.sci.microscopy. My ISP seems to have a problem w/ this one though, casue I lose all the threads in about a week. {p} It works basically the same was as this listserver, only you have the option of reading or not reading messages, rather than having all messages sent to you via email. {p} When i have a question or such, I'll usually post in both places. {br} For example, I posted my questions regarding the TN5500 here and there. I recieved a few responses from there and many more from here. I am not sure, however, how many people know of the newsgroup. {p} Well, thats my two pennies, hope it helps. {p} Ted Claypool {br} EPMA {br} {a href="http://www.rjlg.com"} RJ Lee Group {/a} {/html}
} ... } } } } ... } } } Not interesting, just sad. No matter how good we are at what } we do or how } hard we try, women still are considered second class citizens and our } salaries reflect that. ... } ...
I believe you are correct in many circumstances, but I think you are wrong to make such a conclusion in this case when the "average experience" wasn't reported with the "average salary" ... also not reported with respect to gender; minimum and maximum salaries and minimum and maximum degrees of experience ... (leastwise in this e-mail thread ... I have yet to see the survey). For example. I didn't take part in this survey for my own reasons and I receive a lesser salary too with 20years experience. Personally, I have a lesser salary because I chose this working place and living environment, and didn't believe my salary deserved to be rated with general electron microscopists. My lesser salary would then come under a heading of "aggression for $$$", and I do believe men are more aggressive when it comes to salary demands. Regarding "2nd class citizens" I think things are more complex than you know and not as bad as you imply, but I will agree ... for the same experience salaries should be equal, and I don't think were there yet either.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
} P.S. #1: A surprise (to me) in the survey! The average yearly salary of all } 457 microscopists reported was $51,676. The average yearly salary of all 268 } male microscopists was $55,641 - or up some 7.7 % from the total average. } But, the average yearly salary of all 189 female microscopists was $46,053 - } or down some 10.9 % from the total average. No conclusion suggested! } Interesting?
Years-in-grade (or seniority) and rank have very large and statistically significant impacts on salary. Were the gender data normalized to account for this? If not, then no conclusion is possible.
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
Usually I try to stay out of the "political" discussions, but I would like to put my two cents in on this one.
My initial training in microscopy came from the Royal Microscopical Society in England. One thing which differentiated their approach from more typical US courses was their cross-disciplinary perspective: electron microscopists talked to light microscopists and materials scientists learned new staining approaches from biologists, etc. We all benefit from the diverse postings on the MSA listserver. I agree with Jordi, that headers would be helpful when our mailboxes get too full, but I, for one, learn as much as possible about "what the other guy is doing" so that I can add to my own experience base.
By the way: as an avid "industry watcher", I can tell you with certainty that the current trend in professional societies is toward partnering with sister-disciplines. The fact that our annual meeting is now M&M, not just MSA, should be evidence enough. Let's keep the splintering to a minimum and use headers, instead, to divvy up the info.
I would suggest using a thin 6H SiC specimen in (0001) orietation. 6H - SiC has a lattice spacing of c=1.511 nm , so try to image these lattice fringes in the STEM, but you may find that the collection angle of your objective aperture to be too big i.e. you may have three or four beams, but take several images at slightly different defocus to see what the minimum spacing is between them. Kinematically the (000m) - all m other than m=6n (integer n) beams are forbidden, but they are strong (dynamically) close to and on certain zone axes along the (000n) Kikuchi bands. Good Luck. JOn -- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
The little info I have suggests that Glycerol Borate might be a nearly ideal mounting medium for my purposes. But more info seems hard to find.
It was once available commercially (from ?Glyco?) as "Aqua Resin". Alas, no more. It has been recommended by Dr. McCrone himself. I thought it best to try the list before asking Dr. M. or anyone at MRI.
Can this stuff be made by merely dissolving Borax in glycerin (+ water initially?), or is it more complicated? Does anyone have any recipes? Any tidbits of info will be appreciated.
If you look closely at the address you will see a couple of "-at-" signs. If you pull off the trailing "-at-sparc5.microscopy.com" then you should have a valid address.
I have noticed the same problem on occasion and have so learned to double-check the address before clicking the SEND button. If I forget and get an error, then I know where to look for trouble.
I think Nestor may have explained why this happens. I forget the explanation, and it is not always a problem. So I basically just try to be careful.
Warren
At 09:09 AM 12/7/98 -0800, you wrote: } p.s. everytime I try to reply to the } MicroToday-at-aol.com-at-sparc5.microscopy.com I get a bad address and the } message is thrown back, does anybody know why?
Paula, and others who have read her message as regards the salary survey,
The reply to the listserv:
"Not interesting, just sad. No matter how good we are or how hard we try, women still are considered second class citizens and our salaries reflect that. We're used to, but not happy about, being paid less almost 100% of the time.
Paula :-(
Paula Sicurello"
is an example of sloppy thinking. One piece of summary statistical data does not demonstrate anything about the treatment of a group as there may be systematic factors which explain data such as was cited. After all, there is more data, according to the authors of the survey, not yet put on the listserv due to formatting questions.
After all, those who conducted the survey wrote previously that there were not enough data points to justify the publication of the survey, in their original opinion. The use of small samples to measure a group leads to errors. The fact that those who conducted the survey brought up such a point after having originally said that there was not enough data to publish suggests that they may have been fishing for statements such as Paula's. In my opinion, that is an irresponsible use of survey data.
A major problem today in the media, one which frustrates scientists such as those who pay attention to this list, is the poor understanding of statistics and their use in science, social or natural, held by reporters and the general public. This can lead to poor decision making which can in turn hurt scientific research in terms of both funding and public opinion. As a good example, consider the nonsense about power lines and cancer.
The comment in question here is another example of such weak reasoning. If scientists choose to think these questions through in such a manner, how can we expect the general public to do otherwise?
Doug Darnowski ****************************************************************************** Douglas Darnowski Department of Crop Sciences 384 ERML 1201 West Gregory Drive University of Illinois Urbana IL 61801 work ph: (217) 244-6150 fx: (217) 333-4777 home ph: (217) 356-6606 fx: (217) 356-4454 email: darnowsk-at-staff.uiuc.edu
We routinely use the dull side, not only because of the increased "stickiness" of the membrane to the rough surface, but also because if you put the film on the shiny side and then coat it with carbon as we always do, you have 2 dull sides. It is, thus, more difficult to determine where the sample is when one flips over--particularly if you're doing negative stains. This may not be the case for cryosections in methyl cellulose or epoxy sections, but we always use the dull side for these too to be consistent.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Confocal microscopy offers a good solution to this problem. Also, you might want to investigate the technology offered by Edge Scientific, both their R400 (a full, real time 3D imaging system which can use Phase and DIC) as well as their new Perceptra illuminating system. I will also forward some other information to you under separate cover. Edge can be reached by email at edgesci-at-primenet. In terms of confocal, there are a number of choices, ranging from BioRad to Carl Zeiss, Leica, Olympus, and Nikon. EG&G Wallac seems to be the newest player in the confocal field. For these companies, I suggest you visit either the MSA web site at www.msa.microscopy.com, MicroWorld at www.mwrn.com, or the Microscopy Vendors' Database at www.kaker.com.
Hope this is helpful.
Best regards,
At 05:51 PM 12/7/98 +0300, Savostjanov G.A. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America | | } | home +7 (812) 5100052 |44 Thores av., 194223, St.Petersburg, Russia | } } }
Dear Confocal users, Below is an email I sent to Bio-rad regarding MRC600's and Y2K. I have received no response from them regarding these issues. Does anyone out there know the answers to these issues? Dear Bio-rad, I am the Confocal / Electron Microscopy Core Facility Manager for the Department of Molecular Biology at Princeton University. I am inquiring as to the Y2K status of our MRC600 Confocal Microscope System. We have had this instrument since 1991, and it continues to be a valuable instrument for this laboratory, and my department. We have collected and archived over 150 gigabytes of image data on optical disks with this instrument. This data needs to be retrievable currently, as well as in the future. The instrument is under service contract We are currently running the Comos7.0 program under Windows 95 software. We are also running macro programs out of SOM under COMOS version 6.03. These macro programs are an essential tool for several of my investigators imaging GFP (green fluorescent protein) in living organisms. Without these macro programs to run the instrument, we would be unable to obtain the information required for these experiments. We also run CAS 2.5 and CAS 3.10 for converting images to tiff formats and for creating 24 bit merged color tiff files for export to other programs. I need to know a) if this instrument will continue operating on and after 01-01-2000, b) whether or not previously collected images will be read by the program, c) if we will continue to be able to run macros under the SOM command shell, d) what software and / or hardware upgrades will be required, and when you will have them available?
Sincerely,
Info & Images at http://www.molbio.princeton.edu/confocal/
Joe Goodhouse Confocal / EM Core Lab Manager Department of Molecular Biology Princeton University jgoodhose-at-molbio.princeton.edu 609-258-5432
I have been looking for a binocular phase-contrast microscope during the last year or so. I recently received an A. Daigger & Company, Inc., Lincolnshire, Illinois catalogue which lists a reasonable priced (1095.00) microscope. The microscope is a LABOROCON (TM) with a 5 year warranty. The price seems reasonable, but I have no experience with this product and don't know anyone who has. I will appreciated hearing from anyone who has any experience with this microscope. I would be interested in learning if anyone knows of somone who handles second hand microscopes that I might contact. As I am not a subscriber, please contact me directly at WPTT39A-at-prodigy. com Regards, Bob Davis
Whether this particular study is statistically sound in terms of men's vs. women's salaries, it is well documented (C&E News and others) that this is still a problem in the scientific arena, even though things have improved from years ago. I happen to think it is quite humorous that whenever this issue comes up, in any conversation or whatever, some man or men immediately jump up and start defending themselves by claiming that this is not true, or even worse, ranting about reverse discrimination that they claim to know all about and have experienced! So defensive! I wonder why?
So how does one explain the dominance of men in the field, as evidenced by fewer tenured professors, etc., and I don't know if NIH breaks down its grants this way, but there are certainly many fewer Howard Hughes recipients who are female, and nobel laureates for that matter. I think it is a rational deduction that there is much less respect for women in science, and that despite the changing times, there is still an "old boys network" in place. Surveys in the business field have certainly demonstrated that women are often paid less despite similar qualifications. I'm sorry that more people did not respond to the survey to achieve statistical significance, as to how the results might have changed with more responses.....
I would like to add my whole-hearted agreement. One of the things I value about the MSA listserver is the ability to read postings from, and make contacts with microscopists from a cross-disciplinary range of backgrounds, particularly since one of my own research areas is interdisciplinary in nature.
Gill Bond Dept Materials & Met. Eng. New Mexico Tech
On Mon, 7 Dec 1998, Barbara Foster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } } } Usually I try to stay out of the "political" discussions, but I would } like to put my two cents in on this one. } } } My initial training in microscopy came from the Royal Microscopical } Society in England. One thing which differentiated their approach from } more typical US courses was their cross-disciplinary perspective: } electron microscopists talked to light microscopists and materials } scientists learned new staining approaches from biologists, etc. We all } benefit from the diverse postings on the MSA listserver. I agree with } Jordi, that headers would be helpful when our mailboxes get too full, but } I, for one, learn as much as possible about "what the other guy is doing" } so that I can add to my own experience base. } } } By the way: as an avid "industry watcher", I can tell you with certainty } that the current trend in professional societies is toward partnering } with sister-disciplines. The fact that our annual meeting is now M&M, } not just MSA, should be evidence enough. Let's keep the splintering to a } minimum and use headers, instead, to divvy up the info. } } } Hope this is helpful. } } Barbara Foster } } Consortium President } } {color} {param} 0000,8080,0000 {/param} Microscopy/Microscopy Education } ..Educating microscopists for greater productivity. } } } {/color} 125 Paridon Street Suite 102 Springfield, MA 01118 } } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } } Visit our web site { {http://www.MME-Microscopy.com/education} } } ****************************************************** } } {bigger} {bigger} MME {/bigger} {/bigger} is America's first national } consortium dedicated to } } customized on-site training in all areas of } } microscopy, sample preparation, and image analysis {bigger} . } } {/bigger} } } } } } } } At 08:10 AM 12/7/98 -0700, Marti, Jordi wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } I tend to agree with Chuck. } } } } } } So far the listserver has been a good source of infomation. I think } we } } } would all lose if we splinter into smaller listservers. If anything } I } } } would consider using better (i.e. clearer) Subject Identifiers. For } } } example, one could use headers such as Bio (for Biological), Mat (for } } } Materials ) and Gen. (for general ) subjects. This would make it easier } for } } } a reader to decide if he/she wants to open a given posting. I } believe } } } something like this was proposed by others in the past. } } } } } } We also get much useful information from vendors (I am not a vendor) } and } } } if we were to split this would pose increased problems for both } vendors } } } as well as us microscopists in terms of the time spent searching } and } } } posting messages. } } } } } } In short, I think we would lose more than what we would gain. } } } } } } Jordi Marti } } } } } } } } } ---------- } } } } From: Garber, Charles A. } } } To: MICROSCOPY BB } } } Subject: New listserver } } } Date: Saturday, December 05, 1998 11:49AM } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } Dr. Rik Brydson wrote: } } } ===================================== } } } Some information on a new listserver recently set up. I encourage you } to } } } join Please display in your institution. } } } } } } Many thanks, } } } ================================================ } } } It is not clear to me how this new listserver would in some way } contribute } } } to the objectives as outlined, in ways not possible via the existing } } } listserver sponsored by the Microscopy Society of America. Right now, } if } } } someone has a question, they need post it in only one place, if the } "places" } } } were fragmented, to get the same full coverage, one would have to place } it } } } in more than one place. } } } } } } And for those who felt they wanted to subscribe to more than one } listserver, } } } they are going to be reading the same postings more than one time. } } } } } } I would cast my vote that the system we have "ain't broken" and } therefore we } } } shouldn't try to fix it! } } } } } } Chuck } } } } } } } } } } } } =================================================== } } } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } } } President 1-(800)-2424-SPI } } } SPI SUPPLIES FAX: 1-(610)-436-5755 } } } PO BOX 656 e-mail: cgarber-at-2spi.com } } } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } } } } } } } Look for us! } } } ############################ } } } WWW: www.2spi.com } } } ############################ } } } ================================================== } } } } } } } } } } } } } } }
My two cents - Some of these issues came up when we started an Australasian microscopy listserver a few years ago. I think there has to be some specialisation, otherwise the global system will get clogged with groups in Rekjavik, East Wapping and Te Awamutu announcing local meetings etc. We find a three-level system seems to work pretty well - so we run a server for Canberra, and one for Australasia, and its a commonsense decision whether to send a message to one of these or to MSA. So the MSA server doubles as the global as well as the national US system, because it was first up, filled a need, can handle the traffic, and because Nestor is willing to do the work. For which much thanks, Nestor! The establishment of more national servers outside the US is a natural evolution, if they are dealing with purely regional issues. But trying to split the "world-wide" conversation place into regional groups would be pointless. Probably any schism is liable to be self-healing anyway - methodological postings and general questions of techno-socio-philosophy will tend to be sent to the widest distribution list.
Sally ---------------------------------------------------------------------- Dr Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: GPO Box 475 ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 (0)2 6249 2743 |Australian National Univ. FAX 61 (0)2 6249 4891 |Canberra, Australia 2601 http://online.anu.edu.au/EMU/home.htm
BArbara what antibody are you working with? how are you processing the tissue? what are your results from LM of DAB immunostaining? what dilutions are you currently using with your Abs? these are a few questions needed to be addressed, I would be happpy to help once I have more info
-Mike Rock On Fri, 4 Dec 1998, Barbara Plowman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Listserver! } I am doing some immunohistochemistry and have made two attempts but with no } luck at getting any gold label. My question is whether to use a lower } dilution gold-antibody or extend the incubation from 4 hours to overnight or } both? I am a self-confessed rookie at this, so any information on the subject } would be appreciated . } }
Elaine- if you examine grids by SEM you will "see" there is adifference between the shiny side (which is smoother) than the dull side of a grid (which is rougher) the rougher side has more surface area and thus makes a better contact with films or sections, which in turn keeps them on the grid. -Mike
We have a JEOL 100B (1974) TEM available at no cost. The microscope is operational and was converted to operate at 120kV. Also included will be a second column and HV tank and various other accessories.
Any interested party should contact me as follows:
Paul E. Fischione E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 Phone (724)325-5444 FAX (724)325-5443 e-mail pe_fischione-at-fischione.com
Not a problem a cryostat would make this very easy to section -at- 10 microns you would need to have specimen temperature control available, and have the microtome, knife & anti-roll set to 15-20 deg C colder than the lipstick.
Best Regards
Alan Bright
Bright Instrument Co.Ltd. St Margarets Way Huntingdon PE18 6EB England
Tel No; 01480 454528 Fax No;01480 456031 Email ; Bright-at-dial.pipex.com
-----Original Message----- } From: Barbara Weyn {ingridb-at-uia.ua.ac.be} To: Microscopy-at-Sparc5.Microscopy.Com {Microscopy-at-Sparc5.Microscopy.Com}
Personally, the quantity of information, though somewhat diverse, is perfectsly fine. I agree, be more clear w.r.t. subject headings. I usually delete a handful or more of msg's before opening any if they appear to not be of interest. I also subsc. to about five listservers in other areas, and run then all through the same email account. Roughly 500-750 emails per day on this account alone. Absolutely no problems.
Leave the Microscopy list alone. Much of the Bio-type information has been relevant to my work in Materials-type research, etc.
TJ
__ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3392 [x4] `___/ //\./=/~\====\ (704)509-6622 [Hm] \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
On Mon, 7 Dec 1998, Marti, Jordi wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I tend to agree with Chuck. } } So far the listserver has been a good source of infomation. I think we } would all lose if we splinter into smaller listservers. If anything I } would consider using better (i.e. clearer) Subject Identifiers. For } example, one could use headers such as Bio (for Biological), Mat (for } Materials ) and Gen. (for general ) subjects. This would make it easier for } a reader to decide if he/she wants to open a given posting. I believe } something like this was proposed by others in the past. } } We also get much useful information from vendors (I am not a vendor) and } if we were to split this would pose increased problems for both vendors } as well as us microscopists in terms of the time spent searching and } posting messages. } } In short, I think we would lose more than what we would gain. } } Jordi Marti } } } ---------- } } From: Garber, Charles A. } To: MICROSCOPY BB } Subject: New listserver } Date: Saturday, December 05, 1998 11:49AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Dr. Rik Brydson wrote: } ===================================== } Some information on a new listserver recently set up. I encourage you to } join Please display in your institution. } } Many thanks, } ================================================ } It is not clear to me how this new listserver would in some way contribute } to the objectives as outlined, in ways not possible via the existing } listserver sponsored by the Microscopy Society of America. Right now, if } someone has a question, they need post it in only one place, if the "places" } were fragmented, to get the same full coverage, one would have to place it } in more than one place. } } And for those who felt they wanted to subscribe to more than one listserver, } they are going to be reading the same postings more than one time. } } I would cast my vote that the system we have "ain't broken" and therefore we } shouldn't try to fix it! } } Chuck } } } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: www.2spi.com } ############################ } ================================================== } }
.again, a concurrence. Notes from abroad (from the U.S.) are usually the notes to watch for. :) IN America I've noticed time and again that we want to 'finitize' everything. Hence the reason I initially avoided Electrical Engineering as a course of study. Leave the list as it is, perhaps tighten up the content, subject lines, and make a more strict statement RE: off-topic posts (LIKE THIS ONE!) --i.e. forbid them.
TJ
__ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3392 [x4] `___/ //\./=/~\====\ (704)509-6622 [Hm] \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
On Mon, 7 Dec 1998, Barbara Foster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } } } Usually I try to stay out of the "political" discussions, but I would } like to put my two cents in on this one. } } } My initial training in microscopy came from the Royal Microscopical } Society in England. One thing which differentiated their approach from } more typical US courses was their cross-disciplinary perspective: } electron microscopists talked to light microscopists and materials } scientists learned new staining approaches from biologists, etc. We all } benefit from the diverse postings on the MSA listserver. I agree with } Jordi, that headers would be helpful when our mailboxes get too full, but } I, for one, learn as much as possible about "what the other guy is doing" } so that I can add to my own experience base. } } } By the way: as an avid "industry watcher", I can tell you with certainty } that the current trend in professional societies is toward partnering } with sister-disciplines. The fact that our annual meeting is now M&M, } not just MSA, should be evidence enough. Let's keep the splintering to a } minimum and use headers, instead, to divvy up the info. } } } Hope this is helpful. } } Barbara Foster } } Consortium President } } {color} {param} 0000,8080,0000 {/param} Microscopy/Microscopy Education } ..Educating microscopists for greater productivity. } } } {/color} 125 Paridon Street Suite 102 Springfield, MA 01118 } } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } } Visit our web site { {http://www.MME-Microscopy.com/education} } } ****************************************************** } } {bigger} {bigger} MME {/bigger} {/bigger} is America's first national } consortium dedicated to } } customized on-site training in all areas of } } microscopy, sample preparation, and image analysis {bigger} . } } {/bigger} } } } } } } } At 08:10 AM 12/7/98 -0700, Marti, Jordi wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } I tend to agree with Chuck. } } } } } } So far the listserver has been a good source of infomation. I think } we } } } would all lose if we splinter into smaller listservers. If anything } I } } } would consider using better (i.e. clearer) Subject Identifiers. For } } } example, one could use headers such as Bio (for Biological), Mat (for } } } Materials ) and Gen. (for general ) subjects. This would make it easier } for } } } a reader to decide if he/she wants to open a given posting. I } believe } } } something like this was proposed by others in the past. } } } } } } We also get much useful information from vendors (I am not a vendor) } and } } } if we were to split this would pose increased problems for both } vendors } } } as well as us microscopists in terms of the time spent searching } and } } } posting messages. } } } } } } In short, I think we would lose more than what we would gain. } } } } } } Jordi Marti } } } } } } } } } ---------- } } } } From: Garber, Charles A. } } } To: MICROSCOPY BB } } } Subject: New listserver } } } Date: Saturday, December 05, 1998 11:49AM } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } Dr. Rik Brydson wrote: } } } ===================================== } } } Some information on a new listserver recently set up. I encourage you } to } } } join Please display in your institution. } } } } } } Many thanks, } } } ================================================ } } } It is not clear to me how this new listserver would in some way } contribute } } } to the objectives as outlined, in ways not possible via the existing } } } listserver sponsored by the Microscopy Society of America. Right now, } if } } } someone has a question, they need post it in only one place, if the } "places" } } } were fragmented, to get the same full coverage, one would have to place } it } } } in more than one place. } } } } } } And for those who felt they wanted to subscribe to more than one } listserver, } } } they are going to be reading the same postings more than one time. } } } } } } I would cast my vote that the system we have "ain't broken" and } therefore we } } } shouldn't try to fix it! } } } } } } Chuck } } } } } } } } } } } } =================================================== } } } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } } } President 1-(800)-2424-SPI } } } SPI SUPPLIES FAX: 1-(610)-436-5755 } } } PO BOX 656 e-mail: cgarber-at-2spi.com } } } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } } } } } } } Look for us! } } } ############################ } } } WWW: www.2spi.com } } } ############################ } } } ================================================== } } } } } } } } } } } } } } }
Thanks for telling me what was wrong with the address to Microscopy Today. For those of us who are not real literate with computers the answer was the fact that there were 2 -at- symbols in the address. From now on I will check to make sure there is only one -at-. But does anyone know why there were 2 -at-'s in the address to begin with?
Another comment is to the people to my big sigh... I've heard from several people who thought I jumped the gun by commenting before all the data was presented. That is true but it still does not address the fact the women are paid less than men for the same work. Now, guys, don't get in a huff, but that has been true for years.
Plus to ease the mind of others who feel underpaid because of what the averages were (who like me are paid a lot less than the average), we need to see the total results & find out the demographics of the whole survey. I tried to respond to the survey, but my illiteracy with computers & e-mail prevented me from sending my answers (see above).
Think of it this way, there must be some great paying jobs out there. We just have to know where to find 'em.
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
One solution to your calibration problem is using the MAG*I*CAL TEM Calibration Standard.
The MAG*I*CAL is a TEM calibration standard that performs all of the thr= ee major instrument calibrations for a TEM: image magnification; camera constant for indexing diffraction patterns; and image/diffraction patter= n rotation for relating crystal directions to features in the image. =
MAG*I*CAL consists of an electron transparent cross-sectional TEM sample made from a MBE grown, single-crystal semiconductor wafer. When the calibration structure is viewed in a TEM, it appears as a series of light=
and dark layers where the layer thicknesses are accurately known. The calibrated thickness measurements of these light (silicon) and dark (SiGe=
alloy) layers are based on careful TEM measurements of the {111} lattice=
spacing of silicon which is visible on the calibration sample itself, and=
are supported by x-ray diffraction measurements. The layer spacings are designed so that the sample can be used to calibrate the entire magnification range in a TEM - from 1,000X to 1,000,000X. As the sample = is also a single crystal of silicon, the calibrations requiring electron diffraction information such as the camera constant and image/diffraction=
pattern rotation can also be performed easily and unambiguously. One single calibration sample can therefore be used to provide all three of t= he major TEM instrument calibrations at all magnifications and all camera lengths.
With regard to the traceability and certification of the MAG*I*CAL(TM) calibration sample, each sample is grown on {001} oriented single crystal=
silicon, and all spacings on the sample are directly referenced to the =
cross-sectional (111) lattice spacing of silicon. This spacing is visibl= e by lattice imaging on the sample itself, giving each sample the capabilit= y of being self-calibrating. Each unit comes with a numbered certificate,=
the =
text of which is included below. This certificate has been used for ISO 9000 certification, with the argument that to our knowledge, this is the highest quality TEM sample available anywhere in the world at this time. = =
The MAG*I*CAL (TM) calibration sample consists of sets of thin, nominally=
10 nm alloy layers of Si0.81Ge0.19 alternating with 10 nm pure silicon layers, on a single crystal silicon {001} substrate. These electronic device quality layers were grown by Molecular Beam Epitaxy (MBE) as strained layers, i.e., the alloy layers have a slightly different crystal=
lattice constant, but are strained to conform to the lattice spacing of pure silicon, so that the material remains single crystal. Lattice image= s should therefore be taken in the region of the sample containing no Ge, b= ut other measurements are unaffected. The layer thickness variation across t= he wafer was measured by double crystal x-ray diffraction (DCXRD) mapping as= { 1.0%. =
All four sets of the five thin Si0.81Ge0.19 alloy layers and alternating pure silicon layers (superlattices) were directly calibrated by high resolution transmission electron microscopy (HREM) with the cross-section= al (111) =
lattice spacing of the single crystal silicon substrate, equal to 0.31354= 3 nm [1]. These measurements are also supported by (DCXRD). =
The error in all spacings in the superlattices is one atomic layer:
=A6t =3D +0.3 nm or approximately +3% The larger, nominally 1.0 micron silicon spacings were calibrated against=
these superlattices. The total error across the entire calibration sampl= e is given as:
=A6t =3D + 3%
[1] CRC Handbook of Chemistry and Physics, CRC Press, Inc., Boca Raton,=
Florida 33431
I also have copies of other research papers that have been written by th= e developer, John Mccaffrey, which will provide you with much greater detail. If you have an interest= , please let me know and I'll forward the information to you.
If you require any additional information, please feel free to contact me=
I have a student that would like to try to section his material. Where can he purchase Z-6040?
Thanks
Mike
============================================================ Michael Dunlap lab (530) 752-0284 Facility For Advanced Instrumentation fax (530) 752-4412 9-Hutchison Hall University of California mrdunlap-at-ucdavis.edu One Shields Ave. http://FAI.ucdavis.edu Davis CA, 95616-8594 ============================================================
Hi: I am still looking for a Zeiss 10C. Will pay top $$$ for it. Also, if you need parts for a Philips 300 please let me know. Peter Jordan, EMSI 909 694-1839
} } there is still an "old boys } } network" in place. } } Yes it's nagging social issue, but cripes, maybe we should split into } separate male and female microscopy listservers......... } } } } (just a joke)
I am glad that the sense of humor hasn't totally gone lost...... Thanks Arthur! Thanks mate!
I have followed the discussion about this hot topic with increasing interest. I somehow agree with those saying that you should only believe in a statistic which you have manipulated by yourself.
But even if the survey is not really reliable I am pretty sure that most of us agree that the result does match the general knowledge about discrimiation and that also might be the reason for those frustrated comments. I am happy that the situation of women has increased over the last decades (otherwise I might not be able to post this contribution here), but it is much too early to lean back there is still a lot to fight for.
In my opinion the salary question is just one out of an uncountable number of symptoms, the origin is located much deeper. Just to give some keywords: equal education, who sacrifices the career for looking after kids... and so on...
I remember a conference annoucement on this listserver which said: "Dear Sirs...". OF COURSE I complained and finally it came out that it was because of a misunderstanding and I got a big apologize. Okay. But as long as things like this happen at all, I know that discrimination is present!!
Now to make this contribution seriously scientific: There was a question for literature about negative staining recently. Have a look at: "Has negative staining still a place in biomacromolecuar electron microscopy?", A. Bremer, Ch. Henn, A. Engel, W. Baumeister, U. Aebi, Ultramocroscopy 46 (1992) 85-111
Bettina
*** Bettina Wolpensinger Electron Microscope Unit University of New South Wales Sydney, NSW 2052 AUSTRALIA phone: +612 9385 6390 fax: +621 9385 6400 b.wolpensinger-at-unsw.edu.au http://srv.emunit.unsw.edu.au ***
The Relief Contrast according to Hostounsky, RCH makes it possible to observe objects possessing low natural contrast i.e. possessing almost the same index of refraction as their surroundings. This new contrast enhancing method makes use of interaction of skew monochromatic rays with the aperture of RCH condenser. This device consists of an Abbe-type condenser (numerical aperture 1.2), a relief diaphragm, and a tiltable interference filter. The aperture iris diaphragm of the condenser is controlled by a ring with an aperture scale 0.1 - 1.3, and the relief diaphragm is operated by a lever on the condenser side between 0 and I. The contrast enhancement in the position "O" is minimal and in the position "I" maximal. A knob (also on the condenser side) changes the interference filter tilt which controls the wavelength of the monochromatic light. each condenser is provided with a table showing the wavelength which belong to the scale numbers showing the tilt size. In the set is also contained a wrench serving for insertion and removal of the condenser from its bracket in the substage condenser carrier. The RCH condenser for the relief contrast can by applied only with microscope objectives with magnification 10 : 1 and more. Advantage of the Houstounsky's relief Contrast Method is most esteemed by scientists studying living objects: micro-organisms, cells in tissue cultures, yeast cells, sperm cells, algae, etc. The device is applicable in research laboratory and in medical diagnostic laboratory, ecological and hydrobiological laboratory, and also in school training.
If you need more information do not hesitate to ask.
Price is from US$ 1075 plus postage and handling. attachment for CCD camera and photo available.
Orders: LAMBDA PRAHA Inc. Musilkova 12/448 150 00 Praha 5 Czech Rep. Phone Fax: (420+2) 572 107 60
Keep care and be of good cheer.
Regards
Vratislav Richard Eugene Maria John Baptiste of Bejsak (Bayshark)-Collorado-Mansfeld
Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
Temporally home address: 32 Girrawheen Ave. Kiama NSW 2533 Australia e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz ) phone : 0061 0414 540 465
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
To split listserver into small section do not work, or all section will slowly dissapear. I can see it on Entomology listserver and Buprestidae listserver, first still work well but Buprestidae listserver has zero traffic in this time...
Keep care and be of good cheer.
Regards
Vratislav Richard Eugene Maria John Baptiste of Bejsak (Bayshark)-Collorado-Mansfeld
Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
Temporally home address: 32 Girrawheen Ave. Kiama NSW 2533 Australia e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz ) phone : 0061 0414 540 465
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
I said previously that you should use the (0001) orientation- which is wrong -as this is down the c axis. The orientation of the 6H - SiC you should use is somewhere between [1-100] and the [11-20] directions i.e. with the (000n) planes perpendicular to the beam. Sorry for the mistake.
Jon
-- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
Perhaps you mean glycerin jelly with borax acc. to Fischer? The original recipe was published in Ztschr. f. wiss. Mikr., XXIX, p. 65, 1912.
The main advantage of this solution is, that it is liquid at RT... It's said that it hardens rapidly.
This is the recipe as quoted in Langeron, M.: "Precis de microscopie", 1934, p. 603.
Distilled water 240 g borax (= Na2B4O7): 5g glycerin: 25g gelatin: 40g
Dissolve all at RT. After solution put "some time" in a paraffin incubator. This solution needs to be neutral...
I don't know the Ri of this medium...
Have you considered von Apathy's medium for your purpose?
Hope this helps...
Yvan Lindekens, Belgium.
} The little info I have suggests that Glycerol Borate might be a nearly ideal } mounting medium for my purposes. But more info seems hard to find. } Scott Harden
POSTDOCTORAL POSITION - CELL PHYSIOLOGY I have an opening for a Postdoctoral Fellow in my laboratory . The research is concerned with the cellular and molecular biology of drug transport across the renal tubule and the blood-brain barrier. We use isolated renal proximal tubules, renal cells in culture, isolated brain capillaries, fluorescent substrates and confocal microscopy to follow transport across cells. Our goals are to 1) characterize the fundamental membrane-based and intracellular processes involved, 2) determine how they are controlled by hormones and xenobioitcs, and 3) identify the signal transduction pathways involved. Candidates should have a Ph.D. or M.D. degree, less than 5 years of postdoctoral experience and a background in cellular physiology, membrane transport or renal physiology. This is a non-tenure track position (NIH IRTA Fellow).
A few months ago, there was a thread on Automatic Specimen processors. At that time, we were not interested in purchasing one, but certain circumstances have led us to reconsider. I need some prices on them, and some feed back from anyone who may have purchased one and could provide me with some information on the advantages and/or disadvantages with their product. If people could reply to me directly, I would greatly appreciate. I need this information ASAP (like today), so I can put forth the request to the powers that be.
Quotations and information can also be faxed to the number below.
Cheers,
Susan
Susan Carbyn (Electron Microscopist) Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------- Forwarded by Nancy Zjaba/Americas/NSC on 12/08/98 08:17 AM ---------------------------
Nancy Zjaba 12/08/98 07:29 AM To: B.Wolpensinger-at-unsw.edu.au -at- Internet cc:
Interesting thread on the salary survey results...
I've worked in various settings, not all scientific, and one of the things I have found interesting is the reaction of some men to blatant discrimination against the women they work with. While they have not been directly responsible for the discrimination, they simply sit back and observe silently. They are in no way inclined to alter a situation that benefits them. Logically, why should they?
I think that's a big part of the issue. For discrimination of any kind to end, the members of the favored group must exercise some form of moral conviction to stop the discrimination. Sometimes that happens, sometimes it doesn't.
Nancy Zjaba National Semiconductor (a very good place to work, I must add) South Portland, ME
by node21.frontiernet.net (8.8.8a/8.8.8) with SMTP id IAA94146; Tue, 8 Dec 1998 08:25:06 -0500
Let me put forth a 'possible' (i.e., no data to support this, just observation) explanation - as a vendor who has worked with the microscopy community for a few years, I have a noticed that many more women are entering this profession than men. I believe that I have read that this is true in most science (not necessarily engineering) fields today. If that observation is correct, then perhaps the lower average salary is a result of greater numbers of women new to the field.
Yes, there does seem to be an old boys network at the top, but given the sheer numbers of women entering the field, that can't stay like that forever. I don't want to belittle this if it truly is a case of women doing the same job and being paid less - that certainly is not right - but I think that if my observations are correct, and have contributed to this sampling error - then maybe in the long run it is a good sign, and as these new microscopists gain experience the median salary will rise, and the differential will decrease.
Just a thought, and only my own - not my employers -
------------------------------ Scott Ireland North American Sales Manager Media Cybernetics, L.P. "The Imaging Experts" 716.473.0222 Tel 716.473.8048 Fax 888.691.2492 Pager scott-at-mediacy.com http://www.mediacy.com http://www.optimas.com -----------------------------
-----Original Message----- } From: Dr. Rachel Teitelbaum [mailto:teitelba-at-aecom.yu.edu] Sent: Monday, December 07, 1998 5:26 PM To: Douglas W. Darnowski Cc: microscopy-at-sparc5.microscopy.com
So how does one explain the dominance of men in the field, as evidenced by fewer tenured professors, etc., and I don't know if NIH breaks down its grants this way, but there are certainly many fewer Howard Hughes recipients who are female, and nobel laureates for that matter. I think it is a rational deduction that there is much less respect for women in science, and that despite the changing times, there is still an "old boys network" in place. Surveys in the business field have certainly demonstrated that women are often paid less despite similar qualifications. I'm sorry that more people did not respond to the survey to achieve statistical significance, as to how the results might have changed with more responses.....
maybe some of you guys could help this gentleman out. This is out of my field here. Reply to him directly as he is not yet a member of this list. His email address is:
encosrl-at-tin.it
thanks
} From: "ENCO Srl" {encosrl-at-tin.it} } To: {sdw-at-biotech.ufl.edu} } Subject: A request of information } Date: Tue, 8 Dec 1998 11:50:30 +0100 } X-MSMail-Priority: Normal } X-Mailer: Microsoft Outlook Express 4.71.1712.3 } X-MimeOLE: Produced By Microsoft MimeOLE V4.71.1712.3 } } Dear Ladies and Gentlemen I am an Italian student of physics at } university of Padua (Italy). I am going to work on a thesis about wetting } and adhesion properties of liquid on glass and polymers . I am very } interested on finding correlations between the surface energetical } properties of glass and polymers surfaces determinated by macroscopic } wettability and contact angle measurements and their microscopic structure } studied by spectroscopic technique. At the moment, one argument could be } the problems arising when pharmaceutical products become in contact with } glass or polymeric compounds; some pharmaceutical products in fact can } adhere on glasses or polymers loosing or chancing in that way their } properties. Unfortunately I do not know exactly which technique I have } better use and I can not find any good references about the idea to } correlate wettability to those microscopic technique. On your well } organised web site I could understand you are dealing with similar } problems everyday. Would you mind helping me, please ? I need to know if } it is possible: Where is possible to find some bibliographic } references If someone has already studied the problem of } wettability of glasses and polymers by liquids How to correlate the } microscopic structure and/or chemical composition of glass surfaces to } contact angle measurement. Indeed, please I would like to know what } do you think about these project: composition can changed and so } contact angle and wettability measurement can change . ? ) and } then correlate the changes of contact angle and wettability to different } chemical structure of glasses surfaces . Please may you tell me what you } sincerly think about this project ? My adress is: encosrl-at-tin.it Best } regard and of course thank you very much . Via filande, 13 30038 } Spinea (Venice), Italy.
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
maybe some of you guys could help this gentleman out. This is out of my field here. Reply to him directly as he is not yet a member of this list.
His email address is:
encosrl-at-tin.it
} From: "ENCO Srl" {encosrl-at-tin.it} } To: {sdw-at-biotech.ufl.edu} } Subject: A request of information } Date: Tue, 8 Dec 1998 11:50:30 +0100 } X-MSMail-Priority: Normal } X-Mailer: Microsoft Outlook Express 4.71.1712.3 } X-MimeOLE: Produced By Microsoft MimeOLE V4.71.1712.3 } } Dear Ladies and Gentlemen I am an Italian student of physics at } university of Padua (Italy). I am going to work on a thesis about wetting } and adhesion properties of liquid on glass and polymers . I am very } interested on finding correlations between the surface energetical } properties of glass and polymers surfaces determinated by macroscopic } wettability and contact angle measurements and their microscopic structure } studied by spectroscopic technique. At the moment, one argument could be } the problems arising when pharmaceutical products become in contact with } glass or polymeric compounds; some pharmaceutical products in fact can } adhere on glasses or polymers loosing or chancing in that way their } properties. Unfortunately I do not know exactly which technique I have } better use and I can not find any good references about the idea to } correlate wettability to those microscopic technique. On your well } organised web site I could understand you are dealing with similar } problems everyday. Would you mind helping me, please ? I need to know if } it is possible: Where is possible to find some bibliographic } references If someone has already studied the problem of } wettability of glasses and polymers by liquids How to correlate the } microscopic structure and/or chemical composition of glass surfaces to } contact angle measurement. Indeed, please I would like to know what } do you think about these project: composition can changed and so } contact angle and wettability measurement can change . ? ) and } then correlate the changes of contact angle and wettability to different } chemical structure of glasses surfaces . Please may you tell me what you } sincerly think about this project ? My adress is: encosrl-at-tin.it Best } regard and of course thank you very much . Via filande, 13 30038 } Spinea (Venice), Italy.
Cool lipstick to -20C and cut sections in a cryomicrotome using a good steel knofe.
Patricik Echlin Cambridge
On Mon, 7 Dec 1998, Barbara Weyn wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have to make 10 u sections of lipstick. } Can somebody help us? } } } }
John, When I was testing for Y2K conditions in our labs, I found that many manufactures had not yet tested their own equipment - usually our tests and results prompted vendor action! We each hold the responsibility of performing tests on our tools to ensure that, even when the vendor claims compliance, nothing has been missed. I have developed some basic testing protocols that can be used on PC based equipment (a booklet for $15). I'm a firm believer that systems should be tested both before and after a "Y2K fix" has been put into place. I have seen instances where companies can claim compliance, but when tested, a system can still fail. While I'm not a software guru, and do not claim to have the "fix", I've strongly felt a need to improve the general understanding of the Y2K problem and hope to bring some knowledge and responsibility to the public and industry concerning this serious problem. (Sorry to soapbox).
In a message dated 12/8/98 12:56:40 AM Mid-Atlantic Standard Time, jgoodhouse-at-molbio.princeton.edu writes:
{ { Dear Confocal users, Below is an email I sent to Bio-rad regarding MRC600's and Y2K. I have received no response from them regarding these issues. Does anyone out there know the answers to these issues? Dear Bio-rad, I am the Confocal / Electron Microscopy Core Facility Manager for the Department of Molecular Biology at Princeton University. I am inquiring as to the Y2K status of our MRC600 Confocal Microscope System. We have had this instrument since 1991, and it continues to be a valuable instrument for this laboratory, and my department. We have collected and archived over 150 gigabytes of image data on optical disks with this instrument. This data needs to be retrievable currently, as well as in the future. The instrument is under service contract We are currently running the Comos7.0 program under Windows 95 software. We are also running macro programs out of SOM under COMOS version 6.03. These macro programs are an essential tool for several of my investigators imaging GFP (green fluorescent protein) in living organisms. Without these macro programs to run the instrument, we would be unable to obtain the information required for these experiments. We also run CAS 2.5 and CAS 3.10 for converting images to tiff formats and for creating 24 bit merged color tiff files for export to other programs. I need to know a) if this instrument will continue operating on and after 01-01-2000, b) whether or not previously collected images will be read by the program, c) if we will continue to be able to run macros under the SOM command shell, d) what software and / or hardware upgrades will be required, and when you will have them available?
Sincerely,
Info & Images at http://www.molbio.princeton.edu/confocal/
Joe Goodhouse Confocal / EM Core Lab Manager Department of Molecular Biology Princeton University jgoodhose-at-molbio.princeton.edu 609-258-5432 } }
Since I deal withSEMs, I don't always pay close attention to the LM topics. It would seem, once again that I have been short sighted. Another group in our department needs to remove Epon form a sampe to try decalcifying and reembeding in wax. Is there a way to do this? I thought there was a method mentioned recently.
On Mon, 07 Dec 1998 13:38:17 -0500 Barbara Foster {mme-at-map.com} wrote: } Usually I try to stay out of the "political" discussions, but I would } like to put my two cents in on this one. } My initial training in microscopy came from the Royal Microscopical } Society in England. One thing which differentiated their approach from } more typical US courses was their cross-disciplinary perspective: } electron microscopists talked to light microscopists and materials } scientists learned new staining approaches from biologists, etc. We all
Thanks for that stereotypic view of American Educational Styles. I'm glad you at least interjected "typical" in your comment. I can rest easy now knowing that our center is atypical in its approach to training microscopists here in Georgia. ******************************************** John P. Shields Center for Ultrastructural Research Barrow Hall University of Georgia Athens, GA 30602 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
Technical Staff Position in Materials Characterization/Microprobe Analysis Los Alamos National Lab Nuclear Materials Technology Division Materials Science and Processing Group (NMT-11)
The Materials Science and Processing Group (NMT-11) of the Nuclear Materials Technology (NMT) Division at Los Alamos National Laboratory is seeking a highly motivated scientist interested in the study of plutonium and other actinide materials. The successful candidate will participate in ongoing experiments in the Pit Surveillance, Pit Rebuild, Pit Manufacturing, and MOX Fabrication Programs. The primary experimental focus will be in maintaining and advancing a microprobe and SEM lab to perform failure and investigative analysis on production problems and component failures of actinide bearing samples. The successful candidate will be expected to design, implement, and perform investigative studies to better understand the complex behavior of plutonium metal and alloys in the thermo-mechanically processed, annealed, and aged conditions. The successful candidate will also be required to perform hands-on work with plutonium and other actinides in a glovebox environment. Opportunities exist to participate in other ongoing group and division research.
Required Skills: Interested applicants shall have a strong materials science background with demonstrated experience in electron-beam instrumentation and associated detectors used for materials characterization, e.g., SEM, microprobe, EDS, WDS, TEM, etc. The position requires strong knowledge and experience in the area of phase transformations and identification. Effective oral, written and interpersonal skills are required. The ability to work on multiple programs concurrently is also required.
Desired Skills: Demonstrated work experience in an industrial environment is highly desirable. Demonstrated experience in the handling and processing of radioactive materials in a glovebox environment.
Education: A Ph.D. in metallurgy, materials science, or other physical sciences is required.
Additional Requirements: This position is subject to the requirements of the Personnel Security Assurance Program (PSAP). Candidates invited for interview for this position will be subject to a pre-employment screening check, medical examination and drug test, and must consent to be in the PSAP program at the time of the interview. A willingness to work on materials problems relevant to the safety of the US nuclear weapons stockpile, and the ability to obtain a "Q" clearance which requires US citizenship are required.
For consideration, submit: resume, publications list, cover letter outlining current research interests, the names, addresses, and phone numbers of three references, and one copy of university transcripts to:
Brad G. Storey Los Alamos National Lab P.O. Box 1663, MS: E505 Los Alamos, NM 87545 storey-at-lanl.gov 505-667-0458 phone 505-665-4394 fax Brad G. Storey Materials Science and Processing Group (NMT-11) Mail Stop: E505 Los Alamos National Lab Los Alamos, NM 87545 storey-at-lanl.gov 505-667-0458 phone 505-665-4394 fax 505-996-3129 pager
{bold} {fontfamily} {param} Times {/param} {smaller} Technical Staff Position in Materials Characterization/Microprobe Analysis
Los Alamos National Lab
Nuclear Materials Technology Division
Materials Science and Processing Group (NMT-11)
{/smaller} {/fontfamily} {/bold} {fontfamily} {param} Times {/param} {smaller} The Materials Science and Processing Group (NMT-11) of the Nuclear Materials Technology (NMT) Division at Los Alamos National Laboratory is seeking a highly motivated scientist interested in the study of plutonium and other actinide materials. The successful candidate will participate in ongoing experiments in the Pit Surveillance, Pit Rebuild, Pit Manufacturing, and MOX Fabrication Programs. The primary experimental focus will be in maintaining and advancing a microprobe and SEM lab to perform failure and investigative analysis on production problems and component failures of actinide bearing samples. The successful candidate will be expected to design, implement, and perform investigative studies to better understand the complex behavior of plutonium metal and alloys in the thermo-mechanically processed, annealed, and aged conditions. The successful candidate will also be required to perform hands-on work with plutonium and other actinides in a glovebox environment. Opportunities exist to participate in other ongoing group and division research.
{bold} {underline} Required Skills:
{/underline} {/bold} Interested applicants shall have a strong materials science background with demonstrated experience in electron-beam instrumentation and associated detectors used for materials characterization, e.g., SEM, microprobe, EDS, WDS, TEM, etc. The position requires strong knowledge and experience in the area of phase transformations and identification. Effective oral, written and interpersonal skills are required. The ability to work on multiple programs concurrently is also required.
{bold} {underline} Desired Skills:
{/underline} {/bold} Demonstrated work experience in an industrial environment is highly desirable. Demonstrated experience in the handling and processing of radioactive materials in a glovebox environment.
{bold} {underline} Education:
{/underline} {/bold} A Ph.D. in metallurgy, materials science, or other physical sciences is required.
{bold} {underline} Additional Requirements:
{/underline} {/bold} This position is subject to the requirements of the Personnel Security Assurance Program (PSAP). Candidates invited for interview for this position will be subject to a pre-employment screening check, medical examination and drug test, and must consent to be in the PSAP program at the time of the interview. A willingness to work on materials problems relevant to the safety of the US nuclear weapons stockpile, and the ability to obtain a "Q" clearance which requires US citizenship are required.
For consideration, submit: resume, publications list, cover letter outlining current research interests, the names, addresses, and phone numbers of three references, and one copy of university transcripts to:
Rachel, and others watching this thread: (this is my last message to the thread, if anyone wants to discuss it further, email personally)
} So how does one explain the dominance of men in the field, as evidenced by } fewer tenured professors, etc., and I don't know if NIH breaks down its } grants this way, but there are certainly many fewer Howard Hughes } recipients who are female, and nobel laureates for that matter.
No. Since tenure takes some time to achieve, with a PhD being the usual first step, and since there has been a change in the demographics, with regard to gender, it stands to reason that the breakdown of tenure according to sexual lines may change (indeed, it is changing, as demographics show). Other factors may intervene, such as time taken away from work to raise children, which time detracts from effort towards research and tenure.
As for Howard Hughes and Nobel, consider that those awards are given for doing certain kinds of research and for certain kinds of acheivements. Beyond the factors in the paragraph above, consider that those types of research may not appeal to as many women for some valid reasons. Thus again, an explanation without need to invoke a bogeyman such as "the old boys network."
I will add that I have a BS from Yale, a PhD from Cornell, and am a PostDoc at the U of IL. I am a white man. In the six labs in which I have worked, I have never reported directly to a white man, and in only two cases was a white man the PI (one other lab had a white man as PI, but I was doing some experiments there; he was the only white man on my thesis committee). I have supervised approximately ten undergraduate students, of whom none were white non-hispanic males (there were at other times white male undergrads who I helped with experiments, but none ever reported to me directly). In spite of this, I won an NSF, and have consistently received excellent evaluations and above-average raises, and I have never had a complaint, directly or indirectly, that I had unfairly treated any of these students. My personal experience tells me that you don't have to have people with the same skin color and complement of sex chromosomes to interact with in order to achieve.
} I think } it is a rational deduction that there is much less respect for women in } science, and that despite the changing times, there is still an "old boys } network" in place.
Respect cannot be deduced from salaries alone. It is a subjective factor, and you provide no rational evidence for a connection.
} Surveys in the business field have certainly } demonstrated that women are often paid less despite similar } qualifications.
Not true. Those surveys have generally been shown to be seriously flawed, when years on the job and other relevant factors are considered beyond mere academic qualifications. This topic has been dealt with in the Wall Street Journal in recent months.
} I'm sorry that more people did not respond to the survey } to achieve statistical significance, as to how the results might have } changed with more responses.....
Until you have those responses, it is meaningless to discuss or consider, which is why those results shouldn't have been posted.
I will add further that my experience has been that such complaints as were raised by you and Paula are usually made in an emotional way without strong rational bases (see my criticisms above). Until someone can provide something beyond assertions of a conspiracy, there is no reason to believe in them, as far as salaries go. It would be illogical and counter to the principles on which this nation is founded.
Doug Darnowski
****************************************************************************** Douglas Darnowski Department of Crop Sciences 384 ERML 1201 West Gregory Drive University of Illinois Urbana IL 61801 work ph: (217) 244-6150 fx: (217) 333-4777 home ph: (217) 356-6606 fx: (217) 356-4454 email: darnowsk-at-staff.uiuc.edu ****************************************************************************** Douglas Darnowski Department of Crop Sciences 384 ERML 1201 West Gregory Drive University of Illinois Urbana IL 61801 work ph: (217) 244-6150 fx: (217) 333-4777 home ph: (217) 356-6606 fx: (217) 356-4454 email: darnowsk-at-staff.uiuc.edu
Thank-you, Rachel T. and Scott I., for responding to his issue in a manner that appears to be rational and not emotionally generated. I detest the short, inflaming responses to this list that state emotional responses but serve no useful purpose, even when it comes to sharing an opinion for the purpose of debating.
Regarding the "old boy network": I recently had a friend in the field of archaeology reveal to me that her married friend within her graduate program was the only one denied financial support (and teaching experience) because she is married (no children). It was assumed by influential MALE AND FEMALE advisors in the program that she had less career potential because she might decide to have children before she has established herself in her field professionally. This "old boy network" concept is MORE COMPLEX than just blaming males! Please don't make me feel defensive or guilty by implying so!
I think this string of responses is a waste of our time unless we get some more tangible issues to discuss which MAY TAKE US IN A POSITIVE DIRECTION! As a microscopist without a graduate degree, but a good-paying job in the field, I was surprised that the "average" salary in the field was so high! I'm SURE that graduate degrees have a large influence upon this statistic. (Especially since we have already discussed whether we should be insulted by some of the job postings to the list looking for experienced microscopists who will work for $20,000 salaries.)
Note: Along this line of graduate degrees, I know a LOT more females my age (30ish) who are earning, or have received graduate degrees. I suspect that in 20 years, the salaries will very likely swing to the advantage of females (on average). What was the male/female graduate degree ratio 20 years ago from today?
Thank-you for listening to my longish rant. I hope some productive thoughts result from it.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan USA
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Let me put forth a 'possible' (i.e., no data to support this, just observation) explanation - as a vendor who has worked with the microscopy community for a few years, I have a noticed that many more women are entering this profession than men. I believe that I have read that this is true in most science (not necessarily engineering) fields today. If that observation is correct, then perhaps the lower average salary is a result of greater numbers of women new to the field.
Yes, there does seem to be an old boys network at the top, but given the sheer numbers of women entering the field, that can't stay like that forever. I don't want to belittle this if it truly is a case of women doing the same job and being paid less - that certainly is not right - but I think that if my observations are correct, and have contributed to this sampling error - then maybe in the long run it is a good sign, and as these new microscopists gain experience the median salary will rise, and the differential will decrease.
Just a thought, and only my own - not my employers -
------------------------------ Scott Ireland North American Sales Manager Media Cybernetics, L.P. "The Imaging Experts" 716.473.0222 Tel 716.473.8048 Fax 888.691.2492 Pager scott-at-mediacy.com http://www.mediacy.com http://www.optimas.com -----------------------------
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } So how does one explain the dominance of men in the field, as evidenced by } fewer tenured professors, etc., and I don't know if NIH breaks down its } grants this way, but there are certainly many fewer Howard Hughes } recipients who are female, and nobel laureates for that matter. I think } it is a rational deduction that there is much less respect for women in } science, and that despite the changing times, there is still an "old boys } network" in place. Surveys in the business field have certainly } demonstrated that women are often paid less despite similar } qualifications. I'm sorry that more people did not respond to the survey } to achieve statistical significance, as to how the results might have } changed with more responses..... } } -Rachel }
My LINK EDX quant software (ZAFPB) performs a calculation to determine the significance of the peak quant data. The cut off is 2 Sigma. I have lost my book (i.e someone has not returned it) which gives the formula for calculating sigma. Can anyone help out please? Many thanks Chris
Chris Gilpin Experimental Officer Biological Sciences EM Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 0161 275 5170 Fax +44 0161 275 5171 Chris Gilpin http://www.empgu.man.ac.uk
I should have made it more clear on what it is I was looking for information about. It is a TEM tissue processor and I apologize to those I confused.
Susan
Susan Carbyn (Electron Microscopist) Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
To the list: Think about all that's been said. We are all making good money. Some have more experience, some have less. Some live in towns where the rent goes for $450 a month for a two bed apt. some $1200. We will all put food on our tables, and some of us will soon put presents in the hands of our friends and family. We have hope. We discover. We have so much more than we "deserve". Let's be more thankful for what we have, and share a little with those who don't- regardless of the time of year.
Tracey Pepper Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
Anyone out there know of a nice uniform tungsten etch, preferably that will stop on TiN/Ti or SiO2???? Dry or wet would be fine...any suggestions will be much much much appreciated.
John, When I was testing for Y2K conditions in our labs, I found that many manufactures had not yet tested their own equipment - usually our tests and results prompted vendor action! We each hold the responsibility of performing tests on our tools to ensure that, even when the vendor claims compliance, nothing has been missed. I have developed some basic testing protocols that can be used on PC based equipment (a booklet for $15). I'm a firm believer that systems should be tested both before and after a "Y2K fix" has been put into place. I have seen instances where companies can claim compliance, but when tested, a system can still fail. While I'm not a software guru, and do not claim to have the "fix", I've strongly felt a need to improve the general understanding of the Y2K problem and hope to bring some knowledge and responsibility to the public and industry concerning this serious problem. (Sorry to soapbox).
In a message dated 12/8/98 12:56:40 AM Mid-Atlantic Standard Time, jgoodhouse-at-molbio.princeton.edu writes:
{ { Dear Confocal users, Below is an email I sent to Bio-rad regarding MRC600's and Y2K. I have received no response from them regarding these issues. Does anyone out there know the answers to these issues? Dear Bio-rad, I am the Confocal / Electron Microscopy Core Facility Manager for the Department of Molecular Biology at Princeton University. I am inquiring as to the Y2K status of our MRC600 Confocal Microscope System. We have had this instrument since 1991, and it continues to be a valuable instrument for this laboratory, and my department. We have collected and archived over 150 gigabytes of image data on optical disks with this instrument. This data needs to be retrievable currently, as well as in the future. The instrument is under service contract We are currently running the Comos7.0 program under Windows 95 software. We are also running macro programs out of SOM under COMOS version 6.03. These macro programs are an essential tool for several of my investigators imaging GFP (green fluorescent protein) in living organisms. Without these macro programs to run the instrument, we would be unable to obtain the information required for these experiments. We also run CAS 2.5 and CAS 3.10 for converting images to tiff formats and for creating 24 bit merged color tiff files for export to other programs. I need to know a) if this instrument will continue operating on and after 01-01-2000, b) whether or not previously collected images will be read by the program, c) if we will continue to be able to run macros under the SOM command shell, d) what software and / or hardware upgrades will be required, and when you will have them available?
Sincerely,
Info & Images at http://www.molbio.princeton.edu/confocal/
Joe Goodhouse Confocal / EM Core Lab Manager Department of Molecular Biology Princeton University jgoodhose-at-molbio.princeton.edu 609-258-5432 } }
Many thanks to all those who answered my request for a source of parts for an MT2 microtome. Someone here in Canada is able to help me (thanks, Chris). It's good to know there are so many people still using the old machines.
Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-8759 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Dear Peter, Do you currently have dual stages that you'd like to fit into your system, or are you looking for someone to design them for you? There are a few companies you can talk with (the manufacturer of your SEM would be a place to start), but I've also spoken with Leo Fama, the president of AMT (508) 774-5550, and Mark Reynolds of the Ernst Fjeld Company (978) 667-1416 ext. 10. I've worked with both of these companies to design ultra high precision 5-axis SEM stages for use in the semiconductor industry.
Let me know how you fare! Lisa Montanaro Consultant Microscopy/Microscopy Education ph: (703) 257-7157 fax:(703) 365-2427 e-mail: cmontana4-at-aol.com
In a message dated 12/8/98 4:47:34 AM Mid-Atlantic Standard Time, marienhoff.visitec-at-t-online.de writes:
{ { he Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Dear all,
We want to integrate two separate motorized tables of five axis each into our large chamber SEM. Who knows suppliers of these tables?
Thanks for the info, Yvan. I don't think that's the same stuff, but it sounds interesting anyway. The medium I'm wondering about was described as "glycerin borate... once available commercially as "Aqua Resin"...". My impression is that it's either a compound or a simple solution of glycerin, "borate" (+ water?).
Leonard Corwin kindly provided the following: { { FWIW, Beilstein, system number 38 or 39, p. 519 of Band I: boric acid glycerin triester, glassy yellow mass on heating of glycerin with boric anhydride (1866).} }
I don't know if that's it either, but doesn't sound that hard to make (?) if so.
I've not heard of "Apathy's medium"; will try to find references.
I'm looking for a water soluble medium for permanent mounts of chitinous arthropod appendages. The ideal (?) mountant would; 1) Have a Ri of around 1.46; 2) Have low scatter, unlike gum or gelatin media; 3) Be easy to make (and / or be available commercially). It would also be nice if it was non - fluorescent and shrink little or not at all as it dries... but I realize I can't have everything.
Since the salary survey data concerning microscopists is not available at the moment, nor does it necessarily have the information required to adequately interpret it in required to microscopists qualifications, can the political discussion move off line or to some more relevant listserver. The current discussion seems to have little relevance to microscopy.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
If TEM grids are investigated by SEM on both sides, it will be found that one side is much smoother than the other. One side is very rough. If an epon section is viewed by SEM both sides are found to be rough (Handbook of Epoxy Resins). Now, let us say you have two pieces of extra fine sandpaper. One piece of sandpaper is shiny and slick on one side and rough on the other. You want the best adhesion (no slippage) when putting the sandpapers on top of one another. How would you do that?
At 11:11 AM 08-12-98 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I agree, let's be thankful for what we have, and that many organizations and people have managed to minimize or eliminate any bias related to sex.
Let's also recognize that there is still a problem at some organizations. I personally know of some egregious cases (at another institution where I have friends) that were only recently corrected after the women sued. When compared to male professors with equivalent academic field, rank, years at rank, etc, the women had better scientific publication records and higher citation rates and yet had been consistently given lower raises such that it took salary increases of more than 20% just to bring some of them even with male counterparts with lower publication and citation rankings. Fortunately, these women had objective measures they could use to demonstrate the problem.
What criteria would one use in the field of microscopy to objectively rank workers? What criteria are useful but somewhat subjective and therefor potentially influenced by unconscious biases?
Happy Holidays,
Karen ******************************************************************* Karen L. Wetmore Grycewicz, Ph.D. Museum of Paleontology University of California Berkeley, CA 94720-4780 karenw-at-ucmp1.berkeley.edu *******************************************************************
We have a Zeiss 10A that has been under continuous service contract until this summer. It is in excellent condition, and is still working fine. I assume the department is willing to part with it since we will eventually put a Philips 300 in there. If you're interested, I could ask what they want for it. What 300 parts are you willing to part with and for how much?
Sara (address at end)
On Mon, 7 Dec 1998, Peter Jordan wrote:
} Date: Mon, 07 Dec 1998 21:02:30 -0800 } From: Peter Jordan {emsi-at-pe.net} } To: Microscopy-at-sparc5.microscopy.com } Subject: Zeiss 10C } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi: } I am still looking for a Zeiss 10C. Will pay top $$$ for it. Also, if } you need parts for a Philips 300 please let me know. } Peter Jordan, EMSI 909 694-1839 } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
borax 5 grams dissolve in 240 ml water add 25 ml glycerine add 40 grams gelatine, dissolve with heat and heat until solution slightly thickens remains fluid at room temperature
Pantin says glycerine jelly has an RI of 1.47
Apathys Gum syrup has an RI of 1.52
Picked gum arabic 50 grams cane sugar........50 grams distilled water 50 ml
dissolve over a water bath
add 0.05 grams thymol
said to set quickly as hard as balsam
recipes from "the Microtomist's Vade-mecum" by Bolles Lee 7th edition 1913.....
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
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Yikes Mikes! You mean to say that microscopists make an average of $46,000 for women and $55,000 for men? I wish I knew of someone, male or female, around here making anything near that amount. The school of EM at which I trained in '96-'97 has been closed [ I was in the last class] and a year later the whole EM lab shut down - government 'downsizing' of the VA system I suppose, particularly of redundant pathology facilities? (q.v. past posts on the future of this application). Perhaps a bit of perspective on the viability of employment in this field rather than valid but self-congratulatory carping by those of you still employed would be welcome. Gee, I gotta find the Bubba Micro.list and get me one of them jobs. ;-} Regards to all. Be of good cheer.
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Please see the attachment (announcement of dean search for our college of engineering). -- ============================== Pnina Ari-Gur, D.Sc. Materials Science and Engineering Western Michigan University Kalamazoo, MI 49008 (616) 387-3372 FAX: (616) 387-6517 email: pnina.ari-gur-at-wmich.edu http://www.wmich.edu/cmd/arigur.htm ==============================
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{h3 ALIGN="CENTER"} {!--mstheme--} {font color="#3399FF"} Dean, College of Engineering and Applied Sciences {br} Western Michigan University {!--mstheme--} {/font} {/h3} {/b}
{p} {small} Western Michigan University seeks applications and nominations for the position of Dean of the College of Engineering and Applied Sciences. {/small} {/p}
{p} {small} Western Michigan University is a Carnegie Doctoral I university with 750 FTE tenure-track faculty and an enrollment of 26,500 students, 25% at the graduate level. It is advancing to Carnegie Research II classification, and plans a substantial investment in the College of Engineering and Applied Sciences, including a new physical facility, to accomplish this goal. In addition to its Graduate College and Lee Honors College, Western supports six degree-granting colleges: Arts and Sciences, Haworth College of Business, Education, Engineering and Applied Sciences, and Health and Human Services. These colleges offer 242 academic programs, including 25 at the doctoral and 60 at the master's levels. {/small} {/p}
{p} {small} The Dean of Engineering and Applied Sciences, who will report to the Provost, must possess the vision that will enable the college to reach its fullest potential in teaching, research and the development of industry partnerships, and also the passion to implement this vision. The dean must demonstrate significant leadership experience, knowledge of current and future issues in engineering education, a record of successful participation in sponsored research, an ability to interact effectively with external constituents, strong communication skills, and possess academic credentials that would qualify for an appointment as professor with tenure in one of the College's departments. {/small} {/p}
{p} {small} The individual selected will assume the academic and administrative leadership of a dynamic and growing college offering 21 baccalaureate, ten masters and three doctoral programs. The college's staff, which currently includes 77 full-time faculty, 25 funded staff positions, and dozens of contract staff and graduate student assistants, teaches more than 3000 students and generated $16.5M in contract research in 1997-98. The College has a strong commitment to research and education, which spans a broad spectrum of engineering and engineering technologies. In addition, it pursues major research and training efforts in printing and in the aviation sciences. The College offers courses and programs primarily on Western Michigan University's main campus in Kalamazoo, but also serves as a major resource for off-campus instruction and economic growth at five sites across West Michigan. {/small} {/p}
{p} {small} WMU's main campus is located just off Interstate 94 and U.S. Highway 131 in the southwest Michigan city of Kalamazoo, which is less than three hours by car from both Detroit and Chicago. With a population of 220,000, Kalamazoo County is served by ample air, train and bus transportation and offers an appealing environment for study, employment, culture, entertainment and all-season recreation. {/small} {/p}
{p} {small} For additional information about WMU and the College of Engineering and Applied Sciences, refer to our Website: {i} http://www.wmich.edu/ {/i} {/small} {/p}
{p} {small} Applications and nominations should be directed to {/small} {/p}
{p ALIGN="CENTER"} {small} Dean James W. Schmotter, Chair, {/small} {/p}
{p ALIGN="CENTER"} {small} College of Engineering and Applied Sciences Dean Search Committee {/small} {/p}
{p ALIGN="CENTER"} {small} Haworth College of Business {/small} {/p}
{p ALIGN="CENTER"} {small} Western Michigan University {/small} {/p}
{p ALIGN="CENTER"} {small} 1201 Oliver Street {/small} {/p}
{p ALIGN="CENTER"} {small} Kalamazoo, MI 49008 {/small} {/p}
{p ALIGN="CENTER"} {small} {/small} {/p}
{p} {small} For fullest consideration, applications and nominations should be received before January 31, 1999. {/small} {/p} {b}
{p ALIGN="CENTER"} WESTERN MICHIGAN UNIVERSITY IS AN {/p}
Could somebody please tell me where Wulff net can be downloaded?
Thanks for your help in advance.
Kun Li
Kun Li, Ph. D Institute of Materials Research and Engineering BLK S7, Level 3 Office: BLK S13, #02-13d National University of Singapore Lower Kent Ridge Road Singapore 119260
Hi Marisa! Depending on how much you want to etch, there are two that I've used to de- process semiconductors. Typically I'm shooting to stop on a TiN liner or Si - I use incremental times with these etches and check with a SEM to determine the depth of the etch (top-down and simple cleaved sections are great for this study).
1). Mix a 1:1 solution of NH4OH to H2O2 - both should be reagent grade, and the H202 about 35%. The tungsten will fizz while under attack - experiment with 2-10 minutes depending on your thickness. Use within 2 hours of mixing for best reliability - the H2O2 likes to decompose. 2). Use household bleach (sodium hypochlorite) at 60 degrees C for about the same amount of time as etch #1. You can purchase a reagent grade for more money, but this does the trick too!
I do a dip rinse in a beaker, then hit it with a stream of water, then blow dry with N2. Ultrasonic cleaning is fine if your structure can stand it, but is usually not required.
In a message dated 12/8/98 11:18:05 PM Mid-Atlantic Standard Time, mahmad-at-semiconductor.com writes:
{ { From: mahmad-at-semiconductor.com (Marisa Ahmad) To: Microscopy-at-sparc5.microscopy.com ('MSA listserver')
Anyone out there know of a nice uniform tungsten etch, preferably that will stop on TiN/Ti or SiO2???? Dry or wet would be fine...any suggestions will be much much much appreciated.
Granted that the salary survey data may not be sufficient for statistical purposes, granted that we all make more than we really need (I'm an old Peace Corps person, so please don't even try to convince me that we're poor), granted that there are questions concerning time of entry into the field, years of service, etc., I think that this is an ideal forum for such a discussion. We are, after all, microscopists and this is, after all, a forum for people like us.
Salary inequities between men and women are not confined to our field. There may always be disagreement about what the figures actually PROVE, but I simply find it amazing that people continue to debate that salaries are unaffected by gender. I'm old enough to have watched this debate unfold for a couple of decades and the story remains the same---women protesting that salaries are unequal and some men protesting (with their higher salaries) that everything is fine and that the problem is with the data collection. Please note that I said "some men". Please note also that I am a white, American euro-male on the wrong side of the Political Correctness spectrum. Please note finally that I have NEVER been paid or offered anywhere near close to the average salary quoted for either men or women in this field, even though I have just been offered a position at a considerably higher salary than I make now, after 12 years in the field. It is still considerably below the average posted for female microscopists. Am I complaining? No.
The story I hear is that women are paid less than men in many fields. Some protest that this is because of late entry by women into these fields. So, are there any figures out there for people in their first, fifth, or tenth year in the field, correlated against educational background?
Let's continue to talk about this. It's as relevant as any other microscopy topic. If their truly is unfairness in pay scales, it should be brought to light and fixed by all means possible. IMHO.
Any suggestions on how to prepair tick eggs successfully for TEM ? Poor penetration seems to be the main problem because the cuticle is very hard. I would appreciate your expert advise very much.
Thanks
Susan Cooper Mrs Susan Cooper Electron Microscope Unit Department of Anatomical Pathology University of the Orange Free State P.O.Box 339 (G5) Bloemfontein 9300 South Africa Fax:0027-51-4473222 Tel:0027-51-4053061 E-Mail: GNAPSC-at-MED.UOVS.AC.ZA
Any people involved with TEM or STEM on the list(s) may be interested in this.
SuperSTEM project
Invitation to Interested Parties for Possible One Year Post
With current advances in lens aberration correctors occurring worldwide, the Electron Microscopy and Analysis (EMAG) committee of the Institute of Physics (IOP) is currently investigating the possibility of setting up an analytical TEM/STEM/ PEELS facility for the UK and European microscopy community. The facility is envisaged to consist of at least two machines capable of high resolution imaging and analysis at the sub-Angstrom level and would provide a user service to external researchers in parallel with constant development in instrumentation. From these discussions, the possibility of an initial one year post, most probably located at Daresbury Laboratories in Cheshire, has arisen. This appointment would enable the project to be throughly researched and prepared for funding submission. The SuperSTEM sub-committee is keen to hear from interested parties and potential applicants for this post. In the first instance, candidates are asked to contact Prof. L.M. Brown. (0044 (0)1223 337291, email: lmb12-at-cam.ac.uk)
_____________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
Hallo Frank, what kind of STEM do you have? We use Si lattice images for the magnification calibration at the VG HB501 UX. If you don't get this resolution, you could use something with a large lattice spacing. Gerd
---------------------- Forwarded by Pat Kingman/arl on 12/09/98 07:45 AM ---------------------------
Pat Kingman 12/09/98 08:02 AM To: LI Kun {k-li-at-imre.org.sg} cc:
Hey Randy don't disillusion me. I was just about to arrange a sex change & apply for a green card !!
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Randy Tindall {rtindell-at-NMSU.Edu} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
Discussion on salaries as it relates to experise in microscopy is in my view within the scope of this forum. It is abit off the main line of microscopy/microanalysis per se, but I think of sufficient interest to the community that if someone has a constructive comment or information it should be posted.
However, I think we have had more than enough of the side jokes, puns, etc. for now.
My guess is that most people have had their say already.
I have an interesting technical problem that I'm hoping the folks on this list can help me figure out.
I have a colleague who is trying to embed dissected renal tubules (20 micron diameter x 500 micron length) for immunofluorescence. He would like to be able to "tack down" individual tubules to some kind of substrate, so that he can keep track of which end he's working from, and would like to cross section them (1-4 micron sections) from end-to-end (vs longitudinal sx). We've tried adhering tubules to Thermanox coverslips and making cryotome sections, but the sections of plastic curled up like a scroll. We've tried Aclar film, and plastic embedding using LR Gold and had the same problem. Is there someone out there who has a suggestion? We have 2 concerns, 1) we don't want to heat the specimen above physiologic temps and 2) we want to keep track of orientation so as to have cross sections of the tubules - adherence to some sort of substrate is one possible solution - any alternatives?
Thanks for your help. Doug .................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
Susan Cooper (Anatomiese Patalogie) wrote: } } TEM --- Fixation and processing of tick eggs. } Any suggestions on how to prepair tick eggs successfully for TEM ? Poor penetration seems to be the main problem because the cuticle is very hard. } I would appreciate your expert advise very much. } Phase-partition fixation would be worth a try, often useful for invertebrate eggs. See Zalokar and Erk, 1977, Stain Technology 52:89. "Phase partition fixation and staining of Drosophila eggs."
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
I have been asked about analyzing a silver filled (70%) conductive epoxy to look for evidence of a lubricant or other material on the surface of the silver flakes in the epoxy. The epoxy is exhibiting variations in conductivity and one of the hypotheses is that there is a lubricant or other material present that may be influencing conductivity. There is much more to the story, but I won't go into that in order to keep the message short.
We have done quite a bit of ESCA and SEM work on the bulk material, but believe we may need to look at the material in a different manner. One of the suggestions has been to microtome the sample and do TEM on it. The thought is we may be able to see the lubricant layer surrounding the silver flakes at high magnifications. I am only slightly familiar with TEM (I'm a SEM and Surface Analysis person) and thought I would approach the group and ask about the feasibility of this method or any other suggestions would be most welcome. We have the material available in the cured and uncured states.
Thanks in advance for any suggestions,
John Giles Principal Materials Engineer Honeywell Space Systems
Kun Li wrote: =============================================== Could somebody please tell me where Wulff net can be downloaded? ================================================ I don't know of any place where they could be "downloaded" but they can be purchased from our firm, SPI Supplies. I presume they can be purchased from others, possibly EBS. You can find out details of these and other crystal plane projections on our website or go directly to URL http://www.2spi.com/outlet/project1.html
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
A couple of negative comments have been posted about the creation of the Lemas list, but no similar fuss was raised when the microprobe list was created which also has considerable overlap with the MSA listserver.
There's no harm in also having lists with a somewhat narrower focus. I'll continue to subscribe to all three and suffer some duplication of messages, but I can certainly understand someone with a materials science interest not wanting to wade though all the biological sample prep material which often dominates the microscopy listserver.
There's not much difference between sorting postings by subject in one listserver and subscribing to multiple listservers, except the latter at least offers the option of not receiving mail which isn't of interest. For folks who pay for their mail messages, that might be significant.
We had a similar problem with wasp eggs and posted the question to the list. The discussion is archived at the following url at the "Tips & Tricks" site.
At 11:47 AM 12/09/1998 GMT+2, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I have news about two useful supplements for MSA's middle school (grades 4-8) manual,"Microscopic Explorations":
Warren Hatch is a Portland, Oregon schoolteacher who makes very good microscopy videos for children. They're cheap ($15-30 postpaid, depending on quantity, format, and destination); he makes them as a labor of love. MICRO asked him to make a "custom" tape to match the exercises in "Microscopic Explorations". It's now ready, and it's as good as his other tapes. It's 33 minutes long & it's available in both VHS and PAL. Contact him directly at {whatch-at-hevanet.com} for ordering information.
If you are already using "Microscopic Explorations" you know that it contains copyable student instructions and worksheets. The Lawrence Hall of Science is now completing a full set of these in Spanish. If you could use them, or know someone who could, please let me know.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
some years ago, I did a lot of work on TEM of lepidopteran eggs, which are likewise enclosed in a +/- thick shell. Best fixation was achieved, although not in every single egg, but in most of them, by pricking the eggshell with a tiny glass needle that you make from a glass capillary. Be sure that the needle has a sharp edge by breaking it with a Dumont forceps (size 7).
As a primary fixative, we used a mixture of glutaraldehyde and osmium tetroxide in cacodylate buffer. Fixation was done on ice for 30-60 minutes. After several rinses, post- osmication was performed followed by dehydration and Epon embedment - see H. Fehrenbach (1995) Zool. Anz. 234:19-41 and D. Zissler, K. Sander (1973) W. Roux' Arch. Entw.mech. 172:175-86.
Hope that helps.
Good luck with your work,
Heinz
----------------------------------------------------------------- Dr. Heinz Fehrenbach Institute of Pathology University Clinics "Carl Gustav Carus" Technical University of Dresden
I originally asked about 10 months ago if anyone was aware of a recent = salary survey for microscopists similar to the one that was conducted by = MSA back in 1984. I had been asked by the Vice Pres of Research and the = Personnel Director at the institution I was employed at, previously, = what were the salaries paid microscopists in reply to my asking for an = adjustment based on my responsibilities and years of experience. ( I = found that the earlier survey by MSA was helpful in a salary adjustment = back in 1988, but now was invalid due to its age.) I believe these = surveys serve a useful purpose as it exposes in equities (as has been = brought out here) and also give us a benchmark when hiring new people, = and are helpful to personnel directors in determining wage scales. = Furthermore there is a significant difference in cost of living through = out the US, which surveys address as well as specialties, etc. =20 I know some fields involving microscopy are experiencing a decline in = demand but the demand in some other areas for microscopists trained in = EM and in the new light microscopies with experience in networking and = image processing has grown. These labs are now looking for people with = more capabilities/experience than when I started working in this field = in the mid seventies and administers/personnel directors expect us to = hire them at salaries that do not reflect their skills. Any new, valid = data would be helpful, unfortunately, not enough people responded to = give us current and valid data. Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
Several months ago I was told about a software developer who has some relatively simple "time-lapse" imaging acquisition programs for digital cameras. I have since lost track of the name and source. Does anyone know who this is?
In the discussion of salaries for Microscopists we must include an extremely informative bit of information put out by the US Dep of Labor in 1983. That is a long time ago now, but nevertheless, it needs to be known. At the time of the statistics, Fe salaries were 70 cents or so to M sal of 1.00. This was widely published ( and much screamed about) in every newspaper, etc. The US Dep of Labor from the census divided men and women into 4 groups and averaged their salaries. The four groups: Married Men, Unmarried Men, Married Women, Unmarried Women. The results were as follows: Married Men had the highest salaries. Unmarried Women had slightly, slightly, slightly less salary. It was approximately 6% less. Then there was a considerable drop for unmarried men. There followed a HUGE drop for Married women. Married women made about half of what married men did. The commentary at that time was (and some of that is still true) that 1) Married women consider their income frequently as a supplement to the family income and work part time while raising the children and running the house. Many women still follow this pattern. Also many women still take off several years while children are young, or follow less ambitious career paths for several years. We do need to consider this in the overall picture. In 25 years of working professionally I have watched men VERY closely (fun). To see what they do and how they do it. Generally, not always, men are MUCH more forceful when asking for raises, or making moves to other towns for more power and more money than women. Men will write letters, threaten the department chair with leaving, endlessly scout the scene for better jobs, etc. Women are just now learning this type of behavior. Women who demand do a lot better than those who don't. It will change. It is changing now. There will be different ways of living in the next two generations. It will be exciting and interesting. Bye, hildy
I was going to say something like this, but I couldn't have said it any better so I will just say " Thank You!!" I am primarily a materials science man myself, with secondary interests and occasional assignments in the bio area. I find I never have enough good information and I can deal with overlap.
-- Respectfully, Bob ( Robert G. ) Lawrence Failure Analyst Motorola Phoenix Corporate Research Lab 2100 E. Elliot Rd. MD 508 Tempe, AZ 85284-1806 Phone: 602-413-5848 Fax: 602-413-5934 Pager: 1-800-759-7243 PIN 834-2458
I originally asked about 10 months ago if anyone was aware of a recent = salary survey for microscopists similar to the one that was conducted by = MSA back in 1984. I had been asked by the Vice Pres of Research and the = Personnel Director at the institution I was employed at, previously, = what were the salaries paid microscopists in reply to asking for an = adjustment based on my responsibilities and years of experience. ( I = found that the earlier survey by MSA was helpful in a salary adjustment = back in 1988, but now was invalid due to its age.) I believe these = surveys serve a useful purpose as it exposes in- equities (as has been = brought out here) and also give us a benchmark when hiring new people, = and are helpful to personnel directors in determining wage scales. = Furthermore there is a significant difference in cost of living through = out the US, which surveys address as well as specialties, etc. =20 I know some fields involving microscopy are experiencing a decline in = demand but the demand in some other areas for microscopists trained in = EM and in the new light microscopies with experience in networking and = image processing has grown. These labs are now looking for people with = more capabilities/experience than when I started working in this field = in the mid seventies and administers/personnel directors expect us to = hire them at salaries that do not reflect their skills. Any new, valid = data would be helpful, unfortunately, not enough people responded to = give us that. Thanks, to Don Grimes though for making an attempt. Maybe, = M&M should conduct one, since the only data available is from 1984.
Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
I am going to make the assumption that since you are resorting to TEM you need to resolve extremely detail within the egg. Just in case that is not the situation, I once did a neat set of confocal images of a moth egg, both inside and out, using confocal. We did the outside using reflected light and the inside using autofluorescence. No fixation was necessary and the results were amazingly detailed. Just a thought.
After reading through some of these listserver segregation msgs I have thought of an idea, which I'm sure someone has constructed before, or perhaps not (even possible). However, here it is:
The listserver should allow users to subscribe to a handful of segregated topical specializations. YOu may choose any number of them. However, when posting the listserver should offer the possibility of posting to all groups, or just a select number of them. Each group will automatically serve onto itself(a person responding to materials will only be posted to materials automagically), however, the listserver should be set up such that a user can post to all groups (to avoid duplication, etc. to people have subscr.d to more than one). The listserver may be set up in such a way that if a user wishes to post, he/she would have to list the groups' names in the subject line, or perhaps the first line of the message, according to which groups the message is to be posted. Perhaps a simple code of "abcd" or "xyzt" could be listed in the subject line, which will tell the listserver to post to all groups, or any variation therein for separate groups. This will save resources and resolve the issue of segregating the microscopy listserver...we could even begin to bring other slightly -removed topics into the listserver with absolutely no one complaining (unless the listserver goes nuts).
The net effect is that everyone can subsr. to all groups (or just the ones they want). When a person wants to post with a general request, e.g. corporate, academic, or other general umbrella topics, the msg can be posted to any number in the set of {xyzt...}. Effectivley no one loses content, no one receives duplication, and everyone can post to all groups. (HENCE EVERYONE IS SATISFIED AND RESOURCES ARE SPARED)
..but I'm not sure how a listserver can be set up to do this...I'm sure it has been done somewhere. ...and shouldn't be a complex task to implement.
[ignore the name below, as I'm not quite willing to write the listserver code] __ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3392 [x4] `___/ //\./=/~\====\ (704)509-6622 [Hm] \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
On Wed, 9 Dec 1998, Rick Mott wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } A couple of negative comments have been posted about the creation of } the Lemas list, but no similar fuss was raised when the microprobe } list was created which also has considerable overlap with the MSA } listserver. } } There's no harm in also having lists with a somewhat narrower focus. } I'll continue to subscribe to all three and suffer some duplication } of messages, but I can certainly understand someone with a materials } science interest not wanting to wade though all the biological sample } prep material which often dominates the microscopy listserver. } } There's not much difference between sorting postings by subject in } one listserver and subscribing to multiple listservers, except the } latter at least offers the option of not receiving mail which isn't } of interest. For folks who pay for their mail messages, that might } be significant. } } Rick Mott } rick-at-pgt.com }
My observations after being on 10 or so lists over the years is that you need a critical mass of active subscribers to make the thing self sustaining. Unless you get a steady stream of posts the list is quickly forgotten. This list has it. The microbeam analysis list doesn't. Any more fragmenation of the list makes both parts less able to survive.
My vote--the area is already being covered my a working list, let's not mess with it.
Best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
It seems that cost cutting and space saving have finally started attacking us! Our darkroom, which I admit is rather large, is wanted by another more influential person. It has been suggested that we attach a digital camera to our Hitachi H600 TEM to take over from the film camera we currently use. I have suggested that digital images are not yet able to produce images as good as our film can and that the digital image can not be increased to a much larger size without appreciable loss of resolution. Can any one please help us find a solution as this same influential person also has their beady little eyes on our laboratory! We need to know 1. How easy is it to digitise the TEM without losing the ability to still turn to film. 2. What sort of camera/system should we be looking at? 3. What sort of software is needed? 4. How much do these things cost?
Sarah Ellis
Trescowthick Research Centre Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne 8006 Victoria Australia
I have mixed feelings about another list server but if they evolve in their own separate ways there may be enough of a committed user base to support both. In any event at least it has been discussed in depth here.
My only suggestions are: 1. that, as many have pointed out recently, this is an international list so could people always put at least a basic address and there country of origin on their mail? There are still times when someone is selling kit or wanting local advice and I waste time checking their e-mail addresses etc in the futile hope of guessing where they are. 2. If people do post to more than one list can they do it at the same time so we can see it in the 'address to:' or 'copies to:' headers and then know whether to delete duplicates?
thanks
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
---------- } From: Rick Mott To: microscopy
} Hello all again, } } } It seems that cost cutting and space saving have finally started attacking } us! Our darkroom, which I admit is rather large, is wanted by another more } influential person. It has been suggested that we attach a digital camera } to our Hitachi H600 TEM to take over from the film camera we currently use. } I have suggested that digital images are not yet able to produce images as } good as our film can and that the digital image can not be increased to a } much larger size without appreciable loss of resolution. Can any one please } help us find a solution as this same influential person also has their beady } little eyes on our laboratory! } We need to know } 1. How easy is it to digitise the TEM without losing the ability to } still turn to film. } 2. What sort of camera/system should we be looking at? } 3. What sort of software is needed? } 4. How much do these things cost? }
One possibility is to use photoplates. I don't have any personal experience with these, but my understanding is they are basically digital film. The microscope does not need to be modified. Photoplates are used in the same fashion as film. I understand they have a better dynamic range than a CCD. They are also much higher resolution than a CCD. The down side is cost. A reader costs in the $15,000 USD range I believe, and the plates are close to $100 USD each. Also each image is 25 - 30 megabytes so disk storage and backup for the images can add up in a hurry.
All of the above is from memory of a conversation with someone who had this setup. The numbers may be misremembered. Hopefully someone who actually has bought and used photoplates will respond.
Tom
Thomas Mullarkey Murray email:tm8a-at-virginia.edu Thornton Hall - MSE phone:(804)982-5659 University of Virginia Fax: (804)982-5660 Charlottesville, VA 22903
POSTDOCTORAL POSITION - CELL PHYSIOLOGY I have an opening for a Postdoctoral Fellow in my laboratory . The research is concerned with the cellular and molecular biology of drug transport across the renal tubule and the blood-brain barrier. We use isolated renal proximal tubules, renal cells in culture, isolated brain capillaries, fluorescent substrates and confocal microscopy to follow transport across cells. Our goals are to 1) characterize the fundamental membrane-based and intracellular processes involved, 2) determine how they are controlled by hormones and xenobioitcs, and 3) identify the signal transduction pathways involved. Candidates should have a Ph.D. or M.D. degree, less than 5 years of postdoctoral experience and a background in cellular physiology, membrane transport or renal physiology. This is a non-tenure track position (NIH IRTA Fellow).
_____________________________ Dr. David S. Miller Laboratory of Pharmacology & Chemistry NIH/NIEHS Research Triangle Park, NC 27709
I use a Hitachi H7000 and, although I don't have your urgent requirement, I have looked at the convenience issue of digital photography on a TEM.
My understanding is that you would probably need to insert some sort of fluorescent screen and high resolution digital camera at either the 35mm camera port or STEM detector port position of the microscope. At present this would either involve Hitachi or a specialist company that you could trust. By the time that you have paid for all the specialist kit the numbers I have been quoted don't come much below 30,000 UK pounds. It's probably cheaper to get the system integrated into a new microscope if and when you upgrade or have some spare petty cash you're not using.
However - have you considered a simpler and much cheaper option. Develop TEM film in a simple darkroom (in an emergency any dark ventilated area will do). Then scan negatives with the highest resolution negative/transparency scanner you can get. I would guess that you could pay anything from 2,000 to 10,000 UK pounds depending on what you wanted and this would include scanner, fast computer , storage, software and printer. The added advantage is that from a typical negative and high resolution scanner it should be possible if necessary to scan at much better resolutions than with the best digital camera. I would guess that you would be able to print at up to at least 5x the original negative magnification if you used the right scanner.
This is my master plan because you still have the archival and universal properties of film but most of the convenience of digital plus you could at some later date do a full up-grade. I'm still awaiting money to do this but for the outlay the benefits greatly outweigh the costs.
Good luck and let me know how you get on.
Malcolm Haswell Electron Microscopy School of Health Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: Ellis, Sarah To: Microscopy Listserver
Hello all again,
It seems that cost cutting and space saving have finally started attacking us! Our darkroom, which I admit is rather large, is wanted by another more influential person. It has been suggested that we attach a digital camera to our Hitachi H600 TEM to take over from the film camera we currently use. I have suggested that digital images are not yet able to produce images as good as our film can and that the digital image can not be increased to a much larger size without appreciable loss of resolution. Can any one please help us find a solution as this same influential person also has their beady little eyes on our laboratory! We need to know 1. How easy is it to digitise the TEM without losing the ability to still turn to film. 2. What sort of camera/system should we be looking at? 3. What sort of software is needed? 4. How much do these things cost?
Sarah Ellis
Trescowthick Research Centre Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne 8006 Victoria Australia
by mailhub.dartmouth.edu (8.8.8+DND/8.8.8) with SMTP id IAA30481 for {MICROSCOPY-at-MSA.Microscopy.Com} ; Thu, 10 Dec 1998 08:56:16 -0500 (EST) Message-id: {12687646-at-vixen.Dartmouth.EDU}
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have been digitizing our images since 1995. Our solution to the massive resolution loss in our biological digital images (301K) is to take films as well (fewer) which serve as archives. My experience is you need 3 digitized images at different magnifications to get out the same information contained in one film. Films may be scanned as 3-4MB images and printed on a good 1200 dpi dye-sub printer or printed the old-fashioned way.
by highgate.mluri.sari.ac.uk (8.8.7/8.8.7) with SMTP id OAA20601; Thu, 10 Dec 1998 14:19:55 GMT Message-Id: {199812101419.OAA20601-at-highgate.mluri.sari.ac.uk} Comments: Authenticated sender is {mi596-at-highgate}
Hello Sarah, You've asked an interesting question (at least to me that is).The organisation I work for may be buying an old TEM in the near future which could be upgraded to digital. So I'd be interested in hearing all the responses. For example, do digital cameras really offer a chance for us to get out and even stay out of the dark room? (apologies if this question sounds simple). I know that one would need a slow scan CCD rather than a TV rate digital camera for suitable quality images. I'd be interested in all your thoughts on the subject. Regards
Martin Roe MLURI Craigiebuckler Aberdeen Scotland U.K.
} It seems that cost cutting and space saving have finally started attacking } us! Our darkroom, which I admit is rather large, is wanted by another more } influential person. It has been suggested that we attach a digital camera } to our Hitachi H600 TEM to take over from the film camera we currently use. } I have suggested that digital images are not yet able to produce images as } good as our film can and that the digital image can not be increased to a } much larger size without appreciable loss of resolution. Can any one please } help us find a solution as this same influential person also has their beady } little eyes on our laboratory! } We need to know } 1. How easy is it to digitise the TEM without losing the ability to } still turn to film. } 2. What sort of camera/system should we be looking at? } 3. What sort of software is needed? } 4. How much do these things cost? } } Sarah Ellis } } Trescowthick Research Centre } Peter MacCallum Cancer Institute } Locked Bag #1 A'Beckett Street } Melbourne 8006 Victoria } Australia } } Phone +61-3-9656 1244 } Fax +61-3-9656 1411 } Email s.ellis-at-pmci.unimelb.edu.au } } }
We only use a very small darkroom to develop the film. Basiclly a closet next to the scope. Then we scan the negs at either 600 dpi or 1200dpi, stor the files on CDs and print on good printers. We have gotten addicted to the enhanced information we get by viewing the images on the monitor, with image enhancement and analysis tools at the click of a mouse. For us it seems to be the best solution for now. We want to bypass film eventually but the cost and drawbacks seem too great for now.
Bob Derm Imaging Center U of W
On Thu, 10 Dec 1998, Ellis, Sarah wrote:
} Hello all again, } } } It seems that cost cutting and space saving have finally started attacking } us! Our darkroom, which I admit is rather large, is wanted by another more } influential person. It has been suggested that we attach a digital camera } to our Hitachi H600 TEM to take over from the film camera we currently use. } I have suggested that digital images are not yet able to produce images as } good as our film can and that the digital image can not be increased to a } much larger size without appreciable loss of resolution. Can any one please } help us find a solution as this same influential person also has their beady } little eyes on our laboratory! } We need to know } 1. How easy is it to digitise the TEM without losing the ability to } still turn to film. } 2. What sort of camera/system should we be looking at? } 3. What sort of software is needed? } 4. How much do these things cost? } } Sarah Ellis } } Trescowthick Research Centre } Peter MacCallum Cancer Institute } Locked Bag #1 A'Beckett Street } Melbourne 8006 Victoria } Australia } } Phone +61-3-9656 1244 } Fax +61-3-9656 1411 } Email s.ellis-at-pmci.unimelb.edu.au } }
I have difficulty explaining to people how and when to use the AUTO FOCUS and AUTO STIGMATOR controls on our Philips 515, and more important, when not to. Although I have an instinctive understanding, I find it very hard to put into words. Does anybody have any materials for public use that might be helpful, and can explain to people the background involved?
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Robert writes ... } } } } I have difficulty explaining to people how and when to use } the AUTO FOCUS and AUTO STIGMATOR controls on our Philips 515, } ...
I would explain their use in terms of "intelligence algorythms" ... that is, I've seen "auto mode" work well with certain objects imaged, and how well they work being dependent on symetries assumed and present. Your users should know that for all objects however, there is no better "intelligence algorythm" than the human eye working with the human brain.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
My colleague and I are trying to cut serial sections, but are having a hard time getting the sections to ribbon nicely. She has tried trimming by hand with a razor and on the microtome with glass knives. If anyone has had good luck with their sections ribboning and would be able to give some helpful hints, we would really appreciate it. Thanks.
Jeannine Caesar Department of Neuroscience University of Pennsylvania
You might want to try the following, adhear it to a slice of the 'meaty' portion of cucumber an then section in a cryostat. I did this for orientation and sectioning of gastric biopsies 15 or so years ago.
-- Begin original message --
} } I have an interesting technical problem that I'm hoping the folks on this } list can help me figure out. } } I have a colleague who is trying to embed dissected renal tubules (20 } micron diameter x 500 micron length) for immunofluorescence. He would like } to be able to "tack down" individual tubules to some kind of substrate, so } that he can keep track of which end he's working from, and would like to } cross section them (1-4 micron sections) from end-to-end (vs longitudinal } sx). We've tried adhering tubules to Thermanox coverslips and making } cryotome sections, but the sections of plastic curled up like a scroll. } We've tried Aclar film, and plastic embedding using LR Gold and had the } same problem. Is there someone out there who has a suggestion? We have 2 } concerns, 1) we don't want to heat the specimen above physiologic temps and } 2) we want to keep track of orientation so as to have cross sections of the } tubules - adherence to some sort of substrate is one possible solution - } any alternatives? } } Thanks for your help. } Doug } .................................................................... } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : } : Research Specialist, Principal University of Arizona : } : (office: AHSC 4212A) P.O. Box 245044 : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : } : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): } :...................................................................: } http://www.pharmacy.arizona.edu/exp_path.html } Home of: "Microscopy and Imaging Resources on the WWW" } } }
-- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
To all concerned: I am also in the process of digitizing our tem. We have a Zeus 902 cem, fortunately it has a side mount for a 35mm camera already in place. Even with a very good camera, there must be a loss of resolution compared to film. Electrons exposing the film emulsion are highly resolved, whereas a fluorescent image is inherently less resolved. We have always said that you can see more on the negative than the view screen. Has anyone evaluated different resolution cameras to see at what point pixel array size becomes irrelevant when imaging a fluorescent screen?
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
Since you raise questions that probably are of interest to many people, I will try to provide some information. Please keep in mind, that my company, Soft Imaging System Corp, produces and sells these systems and I might therefore be a little "biased" towards digital imaging. I will also forward your email to our agent responsible for Australia so he can provide you with more specific information.
1. How easy is it to digitise the TEM without losing the ability to still turn to film.
That is usually not a problem. The digital cameras are either side-mounted on a 35 mm port or bottom mounted below the film chamber. in most cases (if not all), you can retain the film camera and take images on negatives if that is required.
2. What sort of camera/system should we be looking at?
This and the following questions require a bit more thought. The answer usually depends on what you want to to. As I mentioned above, there are basically two different types of camera: side-mounted and bottom mounted. One clear differentiation is the resolution and field of view you can get from these cameras. Since the side-mounted cameras are mounted above the viewing chamber, they "see" about the same area as a negative, sometimes more. On the other hand, the bottom mounted cameras usually see only a small part of the negative area. The side-mounted cameras are better suited for applications where field of view is an issue, the bottom mounted ones are better suited for applications where resolution is the most critical.
Let's have a quick look at side-mounted cameras. They come in various types and configurations. The simplest type is a simple TV camera, where the CCD chip is either lens or fiber coupled to a small scintillator that intercepts the beam (removable). This type of camera is great for teaching and finding areas on a sample, as you get a real-time image on a screen, but the low resolution (640x480 for US standards) makes it less desirable for actual image acquisition (but see below, Multiple Image Alignment).
Another type of cameras are digital cameras for side-mount. They come in many possible sizes and configurations. Usually they have a better resolution and a higher dynamic range (12 bit as opposed to 8 bit for TV cameras), but often they require specific frame grabbers, and the frame rate (numbers of images per second) can drop significantly. That does not seem to be to important at first, but try to adjust astigmatism with a camera that delivers less than 10 frames per second. I personally find it impossible to do. I would say, that a good camera must deliver a resolution of at least 4 times TV (i.e., 1280 x 1024), have a dynamic range of at least 12 bit and a frame rate of 10 or more at full resolution. It should also be able to accept exposure times of several seconds for those darkfield images, which proabably means cooling the camera.
Then there are specialty cameras: high sensitivity cameras, extremely fast cameras, etc. Those can be used, but they are normally not required for "normal" TEM.
Now come the bottom mounted cameras. Almost all of them are fiber coupled for better sensitivity. They are usually square and come in 1Kx1K and 2Kx2K chips. The 2K chips are MUCH more expensive than the 1K chips. They ususally do not have the readout speed of side-mounted cameras, and are basically replacements for negatives. Their lower read out frequency allows better digitization and they are frequently digitized to 14 bit or better. they do only see a part of the negative, and together with the lower readout speed, it can be hard to find the correct area. Most of them, however, have some features (binning, etc.) that allows faster image repetition rate at a reduced resolution. these cameras are often liquid cooled and either hook up to the TEM cooling system or have their own chiller.
Of course you can have both cameras attached at the same time. That way you can pick what camera to use depending on the application.
3. What sort of software is needed?
That again depends on what you want to do. As a minimum I would think, that you need image acquisition from the camera you select (obviously), some gray scale manipulations, non-destructive overlays for documentation, an image database for archiving and retrieving (should be network capable and compatible with other databases), printing capabilities that include Microscopy specific things like scale bars, true magnification, etc. The software should be able to maintain calibration data for different camera setups, if possible communicate with the TEM, and, again if possible, allow for motorized stage control. Important for TEM is also online shading correction to compenstate for fluctuations in the scintillator and illumination, and real-time contrast maximization. This is especially important for 12 bit cameras, as you may have to look for the "right" 8 bits to display and illuminate the sample all the time. Auto-contrast cuts down on the dose. Depending on how you work, a single screen system (with the operating system and the images sharing one monitor) or a dual screen system (with the images displayed on a separate monitor) is best. If you want to use the digital system for astigmatism correction, you need a real-time FFT feature. If you need particle detection Fourier filters, Programming, etc., this should be available also, but is not really necessary for simple image acquisition. You want to keep this option open, though, by selecting software that offers an upgrade path.
4. How much do these things cost?
Ahh, the most important question. The answer is: From anywhere from several thousand $US to hundreds of thousands of $US, depending on manufacturer, quality, software, hardware, cameras, and your demands. I know, this answer is not very satisfying, but it is just as impossible to answer as the question: how much does a car cost. As I said, I will forward your email to our person in Malaysia, and he can answer those more specific questions.
Multiple Image alignment: This is a way to overcome resolution limitations of cameras. One simply takes a series of images that are displaced but overlapping, and the software reconstructs a larger image by montaging the single images. Best done with a motorized stage and controlled by software, but also possible by hand. This can give you very high resolution images (e.g., 3Kx3K) from a low resolution camera. The price you pay for this is that you have to take an image series instead of a single image and the processing takes some time (seconds).
Whew, that turned out to be longer than I had intended. Aplogies to anybody who thinks it is too long.
If you have further questions, please contact me through email or call me. you can also check out our website at
http:\\www.soft-imaging.de
Finally: I do not claim that this is complete. There are many more issued that need to be addressed, but I did not want to make it even longer.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
} } ---------- } From: Ellis, Sarah[SMTP:s.ellis-at-pmci.unimelb.edu.au] } Sent: Wednesday, December 9, 1998 10:18 PM } To: Microscopy Listserver } Subject: Digitization of the TEM } } Hello all again, } } } It seems that cost cutting and space saving have finally started attacking } us! Our darkroom, which I admit is rather large, is wanted by another more } influential person. It has been suggested that we attach a digital camera to } our Hitachi H600 TEM to take over from the film camera we currently use. I } have suggested that digital images are not yet able to produce images as good } as our film can and that the digital image can not be increased to a much } larger size without appreciable loss of resolution. Can any one please help } us find a solution as this same influential person also has their beady } little eyes on our laboratory! } We need to know } 1. How easy is it to digitise the TEM without losing the ability to still } turn to film. } 2. What sort of camera/system should we be looking at? } 3. What sort of software is needed? } 4. How much do these things cost? } } Sarah Ellis } } Trescowthick Research Centre } Peter MacCallum Cancer Institute } Locked Bag #1 A'Beckett Street } Melbourne 8006 Victoria } Australia } } Phone +61-3-9656 1244 } Fax +61-3-9656 1411 } Email s.ellis-at-pmci.unimelb.edu.au } }
Disaster struck our lab recently when the recipe for Reynolds PbCt got tossed, someone made off with our EM book, and my hard drive got wiped. All in one month! My attempt to recreate from memory failed, judging from the disgusting sections I subsequently observed in the TEM. We usually make up 50mLs at at time, please help!
Thanks,
Peggy Brannigan
EM Lab Floral and Nursery Plants Research Unit, National Arboretum USDA, Beltsville, MD. USA 20705
} Hello all, } } My colleague and I are trying to cut serial sections, but are having a } hard time getting the sections to ribbon nicely. She has tried trimming } by hand with a razor and on the microtome with glass knives. If anyone } has had good luck with their sections ribboning and would be able to give } some helpful hints, we would really appreciate it. Thanks. } } Jeannine Caesar } Department of Neuroscience } University of Pennsylvania
Sections don't ribbon when the top and the bottom of the trapezium is not smooth and parallel. Sharp edges (not raggedy) are important. If they are not parallel, they don't connect to form a ribbon. Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
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The company I work for (Compix) has time-lapse image analysis software that supports digital cameras (Hamamatsu, Spot, Photometrics, but sorry not Polaroid yet!). It is relatively inexpensive ($2K) and easy to use. It is being used mostly for DIC/GFP applications. E-mail me if you are interested in more details or phone my office, 520-749-2070 or our main office 800-393-2526.
Tim Bruchman
Western Regional Sales Manager Compix Inc. Imaging Systems
R-Brooks Corl wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Several months ago I was told about a software developer who has some } relatively simple "time-lapse" imaging acquisition programs for } digital cameras. I have since lost track of the name and source. } Does anyone know who this is?
--------------82E464E000FDAFF4FEB219FF Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Tim Bruchman Content-Disposition: attachment; filename="vcard.vcf"
begin: vcard fn: Tim Bruchman n: Bruchman;Tim org: Compix Inc. adr: 11934 E. Makohoh Trail;;;Tucson;AZ;85749;USA email;internet: timbruc-at-azstarnet.com title: Western Regional Manager tel;work: (800) 393-2526 tel;fax: (520) 749-7999 tel;home: (520) 749-2070 x-mozilla-cpt: ;0 x-mozilla-html: FALSE version: 2.1 end: vcard
In response to the original inquiry, I have worked with 2 imaging systems. One uses an external camera (Kodak Mega +) focused on a phosphor screen. It is mounted on the bottom of the column. The film camera is available. It's ok for lower mag ( {x500K) work but suffers in low light applications like HRTEM. It also suffers from spherical aberration although most users don't notice. Can't really give you a cost figure because it was a package deal. It is a good general use system for our multi user facility. Cost aside though I would not replace it with a similar system. The other system I have used was Gatan model 794. This system was quite impressive. The CCD is in the beam but can be retracted to allow the use of film. Viewing area is about the same with both cameras (~1cm dia). Due to cooling of the detector the Gatan camera has a couple of issues that make me less comfortable putting it into a multi user facility. I little interlocking could cure this. Depending on options the 794 is in the 80-90K$ range. Now all of that being said, while digital images will get the data. When I make presentations to people sitting around large thick oak tables. I want photos. Eye wash counts as much as data to some execs. To scanners: if you keep a dark room & print 8x10s you can scan them on a few hundred dollar flatbed scanner & do pretty well. I was suprised at the results. Of course the files can be huge (but cheap). As long a this subject is up. Has anyone had any experience with the Juji FDL 5000 system. Seems it consist of a secret imaging plate that can be taken from the instrument, electronically scanned by their hardware, erased & re used. It's not clear from the literate if it uses your existing camera or replaces your film camera but they are claiming that you get the same viewing area. (real short coming of all other digital systems I've seen)
Disclaimer: I have no business interest the mentioned companies. These are just my thoughts today.
Bruce Brinson Rice U.
When you travel at the speed of light, you don't need a review mirror.
I'm not completely up to date on the new digital technologies, but I concur with the person who advised capturing images on film and scanning them in, at least as far as the TEM is concerned. Film processing takes little space, unless you're really shooting a lot, and scanners are a lot cheaper than decent digital capture systems.
Also, I have yet to see a TEM digital capture system that can generate really useful images, i.e. publication quality. As I said, however, things may have changed.
SEM digital systems are much better, in my experience, and I assume this is because they literally take over the signal capture and image generation electronics of the scope. TEM's, of course, work completely differently.
Images on film are still the highest resolution, most archival, platform independent, and cheapest form of data storage. I do believe, however, that the printing side of photography can now be safely replaced by reasonably priced scanners and printers. Even cheap inkjet printers now are yielding surprisingly good images. These things and Adobe Photoshop are quickly making printing darkrooms a thing of the past for the vast majority of purposes.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
I am attempting to gather info on the Zeiss Axiotron 2 UV optical microscope. I would appreciate hearing from any users concerning its approximate cost, ease of use, and applications.
We are a semiconductor Fab making gallium arsnide 4 inch wafers. Our application for the Axiotron is to determine if 0.5 micron wide optical photoresist lines, which are about 1 micron deep, are fully exposed and opened. This would be in a production environment, where our operators would be inspecting hundreds of these wafers a week. We can successfully make this determination using our Hitachi 4500 FESEM, but this is of course not practical for production.
Please feel free to reply to:
Daniel Wilcox Senior Eng. M/A-COM Fab II 1-888-276-7975, x5847
Sarah- I have been through this discussion many times with current and former employers. First feel fortunate the bean counters are requesting the change. You will find the cost of digital imaging will cost more than the current value of your TEM. Try to get the slow scan CCD digital camera, the TV or video rate cameras are not acceptable for image archiving. These slow scan CCD cameras and their software usually cost $50-60,000 US. Gatan and Dage both make nice systems. I'm sure there are others. get the most for your money, buy the best technology, it will be obsolete sooner than you wish. Working prints can be printed on a 300-1200 DPI laser jet, and look fine, high quality photograde prints are usually printed on a Dye-sublimation printer, kind of spendy, but again well worth the invesment. And you feelings on film vs digital images are correct, You can always go back and use TEM film for publication images if needed. good luck & have fun shopping -Mike
On Thu, 10 Dec 1998, Ellis, Sarah wrote:
} Hello all again, } } } It seems that cost cutting and space saving have finally started attacking } us! Our darkroom, which I admit is rather large, is wanted by another more } influential person. It has been suggested that we attach a digital camera } to our Hitachi H600 TEM to take over from the film camera we currently use. } I have suggested that digital images are not yet able to produce images as } good as our film can and that the digital image can not be increased to a } much larger size without appreciable loss of resolution. Can any one please } help us find a solution as this same influential person also has their beady } little eyes on our laboratory! } We need to know } 1. How easy is it to digitise the TEM without losing the ability to } still turn to film. } 2. What sort of camera/system should we be looking at? } 3. What sort of software is needed? } 4. How much do these things cost? } } Sarah Ellis } } Trescowthick Research Centre } Peter MacCallum Cancer Institute } Locked Bag #1 A'Beckett Street } Melbourne 8006 Victoria } Australia } } Phone +61-3-9656 1244 } Fax +61-3-9656 1411 } Email s.ellis-at-pmci.unimelb.edu.au } }
Jeannine- I remember an old trick using the glue adhesive from double stick tape, solvated in acetone, then allow exess acetone to evaporate off, once the appropriate consistancy is obtained use this "glue on the bottom edge of the block face. ribbons are easy to cut, but sometimes difficult to separate, don't cut too many at once. -Mike Thu, 10 Dec 1998, Jeannine Caesar wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } } My colleague and I are trying to cut serial sections, but are having a } hard time getting the sections to ribbon nicely. She has tried trimming } by hand with a razor and on the microtome with glass knives. If anyone } has had good luck with their sections ribboning and would be able to give } some helpful hints, we would really appreciate it. Thanks. } } Jeannine Caesar } Department of Neuroscience } University of Pennsylvania } } }
We have a new em facility that was built without a darkroom. Our two main TEM's both have slow scan CCD systems for acquisition and TV rate systems for set up.
At 16:18 1998-12-10 +1100, you wrote: } Hello all again, } } } It seems that cost cutting and space saving have finally started attacking } us! Our darkroom, which I admit is rather large, is wanted by another more } influential person. It has been suggested that we attach a digital camera } to our Hitachi H600 TEM to take over from the film camera we currently use. } I have suggested that digital images are not yet able to produce images as } good as our film can and that the digital image can not be increased to a } much larger size without appreciable loss of resolution.
This is probably still true, just. Low magnification pictures are a problem (8K on the microscope gives us a digital image of x80K. You have to remember that the CCD sees what the small focusing screen looks at. At high mags the maximum final useable image is around 10-20x the indicated microscope magnification.
} Can any one please } help us find a solution as this same influential person also has their beady } little eyes on our laboratory! } We need to know } 1. How easy is it to digitise the TEM without losing the ability to } still turn to film.
Easy. Our digital cameras bolt onto the bottom of the column or it is possible to fit a camera side entry, above the viewing screen. In both cases the film camera is still intact. The CCD camera's are cooled, however, so more care has to be taken to ensure as good a vacuum as possible in the camera chamber.
} 2. What sort of camera/system should we be looking at?
You need both a slow scan CCD and Intensified TV camera. We purchased ours from Gatan.
} 3. What sort of software is needed?
We run with Digital Micrograph. The CCD control is integrated within it.
} 4. How much do these things cost?
Around $100K for a 1Kx1K CCD. The larger the CCD the better!
Regards
Alan W Nicholls, PhD Research Electron Microscopist Manager - Electron Microscopy Laboratory
Mailing Address: Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
When I started working as a junior histology tech in a hospital lab, many years ago in England, I noticed that in all the hospital labs I knew of all junior techs were female and all senior techs were male. I fled back to university research labs before I could be forced to have the operation that seemed to be required to climb the ladder.
Lesley Weston.
On Tue, 8 Dec 1998, Scott Ireland wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Let me put forth a 'possible' (i.e., no data to support this, just } observation) explanation - as a vendor who has worked with the microscopy } community for a few years, I have a noticed that many more women are } entering this profession than men. I believe that I have read that this is } true in most science (not necessarily engineering) fields today. If that } observation is correct, then perhaps the lower average salary is a result of } greater numbers of women new to the field. } } Yes, there does seem to be an old boys network at the top, but given the } sheer numbers of women entering the field, that can't stay like that } forever. I don't want to belittle this if it truly is a case of women doing } the same job and being paid less - that certainly is not right - but I think } that if my observations are correct, and have contributed to this sampling } error - then maybe in the long run it is a good sign, and as these new } microscopists gain experience the median salary will rise, and the } differential will decrease. } } Just a thought, and only my own - not my employers - } } } ------------------------------ } Scott Ireland } North American Sales Manager } Media Cybernetics, L.P. } "The Imaging Experts" } 716.473.0222 Tel } 716.473.8048 Fax } 888.691.2492 Pager } scott-at-mediacy.com } http://www.mediacy.com } http://www.optimas.com } ----------------------------- } } -----Original Message----- } } From: Dr. Rachel Teitelbaum [mailto:teitelba-at-aecom.yu.edu] } Sent: Monday, December 07, 1998 5:26 PM } To: Douglas W. Darnowski } Cc: microscopy-at-sparc5.microscopy.com } Subject: the old boys network } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } So how does one explain the dominance of men in the field, as evidenced by } fewer tenured professors, etc., and I don't know if NIH breaks down its } grants this way, but there are certainly many fewer Howard Hughes } recipients who are female, and nobel laureates for that matter. I think } it is a rational deduction that there is much less respect for women in } science, and that despite the changing times, there is still an "old boys } network" in place. Surveys in the business field have certainly } demonstrated that women are often paid less despite similar } qualifications. I'm sorry that more people did not respond to the survey } to achieve statistical significance, as to how the results might have } changed with more responses..... } } -Rachel } } } } }
Try using insects with lots of setae. These usually give auto controls troubles, but can be dealt with manually. I've found dealing with these structures helpful in showing when to use manual and how, and when to use auto. Setae are also good subjects for training folks in the use of gamma controls.
Phil
} I have difficulty explaining to people how and when to use the AUTO FOCUS } and AUTO STIGMATOR controls on our Philips 515, and more important, when } not to. Although I have an instinctive understanding, I find it very hard } to put into words. Does anybody have any materials for public use that } might be helpful, and can explain to people the background involved? } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: | } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } | University of Reading {University internal extension 7867 | } | Whiteknights Fax +44 (0) 118 9750203 | } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } | England URL: http://www.reading.ac.uk/~spsolley | } +------------------------------------------------------------------------+
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net
Dear Pedro, There is a useful section on electron scattering factors in the International Tables for X-Ray Crystallography. I don't know the volume number, but it's not the one with the space group tables, and the section is written by John Cowley.
While you won't find Doyle-Turner type parametrization of the scattering factors in this section, the factors are tabulated as a function of scattering angle, so you might be able to generate such parameters using a fit.
I have a web page which may be useful for transforming from the Doyle-Turner scattering factor parameters to equivalent parameters for the potential. It is: http://www.numis.nwu.edu/Staff/wharton/channeling.html
Good luck Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
mailto:wharton.sinkler-at-anlw.anl.gov
---------- } From: Joao Pedro Esteves De Araujo {Joao.Pedro.Esteves.de.Araujo-at-cern.ch} } To: sinkler-at-apollo.numis.nwu.edu } Subject: Doyle-Turner potential parameters... } Date: Thu, Dec 10, 1998, 8:27 AM }
} Dear Sir } I hope you can help me. } I am trying to find the Doyle-turner potential parameters for } Yttrium (Y). . Unfortunately in their original paper (Relativistic } Hartree-Fock X-ray and Electron Scattering Factors, P.A. Doyle and P.S. } Turner, Acta Cryst., A24, 390, 1968. ) these parameters were not } calculated. } Do you know some more recent reference where I could find this? } Have you some suggestion how to find this information? } I thank you in advance } Yours Pedro } } } } }
I am writing a paper and I need some explanations of field emission guns and why the resolution is so much better when using this gun in a low-voltage SEM.
I am writing a paper and need some basic information on how the field-emission gun works and why the resolution is so much better with this gun in a low-voltage SEM. TIA Tina Schwach
} Disaster struck our lab recently when the recipe for Reynolds PbCt got } tossed.
Here is the recipe (to make about 17ml):
Weigh out 0.44g lead nitrate and 0.59g of sodium citrate into a small glass bottle. Add 10ml glass distilled water, securely fix the cap and shake (by hand) violently for about 1 min. Transfer to a mechanical shaker and allow to shake for a further 30 min. This mixture should by then be white. To this mixture add 2.6ml of freshly made-up 0.1M sodium hydroxide and gently agitate until the cloudiness disappears, then add a further 4ml of distilled water. NB. If any cloudiness persists discard and make it up again. Some people use this diluted up to 1000x with 0.01M NaOH (the original Reynolds reference has more details) but we use it as made up above.
We store this stain in 3ml aliquots in Eppendorff tubes in the refrigerator at about 4C and it keeps for ages.
I hope this puts you on track again.
Regards
Robin
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
We are looking for a cell-chamber to make time lapse recordings of cells. Can someone tell me about experiences made with the FCS2 chamber from Bioptechs? Since we want to make long time recordings focal stability is of special interest for us. If soemone has made expiriences with other systems I also would be interested.
Hi Margaret I found my old book of recipes which includes Pb Cit. It's a few years since I made any as I'm now in a materials em lab but here's how we used to make 50ml:
1.33g lead nitrate 1.76g sodium citrate 30ml boiled distilled water
Mix in a 50ml volumetric flask, shaking vigorously for 1 min. Leave to stand for 30mins with intermittant shaking. Add 8ml of 1M sodium hydroxide (freshly mixed in boiled distilled water). Make up to 50ml with boiled distilled water and mix by inversion.
I hope this works. Best wishes Nikki
Nikki Bock EM Technician Dept. Materials Engineering University of Nottingham Nottingham NG7 2RD (0115) 9513759/9513871 Email: emznjb-at-hermes.nottingham.ac.uk
Try trimming the block with vertical leading face and sides; only the trailing face keep the usual shape in order to avoid flexure of the block during sectioning (This method is described by Fahrenbach 1984. You can also use a solution of mounting media and toluene mixed 1:1 to coat the leading face facilitating the forming of the ribbon. This procedure gave me very good results.
Gary. NTNU-Trondheim Norway.
On Thu, 10 Dec 1998, Elaine Humphrey wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hello all, } } } } My colleague and I are trying to cut serial sections, but are having a } } hard time getting the sections to ribbon nicely. She has tried trimming } } by hand with a razor and on the microtome with glass knives. If anyone } } has had good luck with their sections ribboning and would be able to give } } some helpful hints, we would really appreciate it. Thanks. } } } } Jeannine Caesar } } Department of Neuroscience } } University of Pennsylvania } } Sections don't ribbon when the top and the bottom of the trapezium is not } smooth and parallel. Sharp edges (not raggedy) are important. If they are } not parallel, they don't connect to form a ribbon. } Elaine } } } Dr. Elaine Humphrey } Biosciences Electron Microscopy Facility } University of British Columbia } 6270 University Blvd, mail-stop Botany } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-unixg.ubc.ca } } }
The recipe for Reynold's lead stain is as follows:
Add 1.33g lead nitrate Pb(NO3)2 to 1.76g tri-sodium citrate Na3C6H5O7.2H2O in a 50ml plastic (polymethylpentane) volumetric flask and add 30ml boiled, cooled, CO2 free distilled water. Invert the stoppered flask occasionally for 30 minutes and then add 8ml CO2 free 1N NaOH and mix till clear. Dilute to 50ml and leave to stand overnight before use. Store in refrigerator.
Dan Hill Department of Biochemistry Cambridge University UK
On Thu, 10 Dec 1998, Margaret Brannigan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Disaster struck our lab recently when the recipe for Reynolds PbCt got } tossed, someone made off with our EM book, and my hard drive got wiped. } All in one month! My attempt to recreate from memory failed, judging from } the disgusting sections I subsequently observed in the TEM. We usually } make up 50mLs at at time, please help! } } Thanks, } } Peggy Brannigan } } EM Lab } Floral and Nursery Plants Research Unit, National Arboretum } USDA, Beltsville, MD. USA 20705 } } (301) 504-6097 } } } }
we're having problems Au coating cheese samples for SEM observations. They ared freezed and then coated. While being coated, after only a couple of minutes, they get caramelized (does this word exists?). It looks like a film of caramel is produced on the surface. Since we have now experience and no idea about this type of materials we would like to hear any opinion about coating conditions: should we Au coat them at high pressure for a short time, or low pressure and longer time? what makes this caramel appear?
Please, let us now if any of you have done something like this before. Thanks in advance.
Silvia Montoro Centro Regional de Investigacion y Desarrollo Guemes 3450 3000 Santa Fe Argentina csedax-at-arcride.edu.ar
Good morning: We are also transitioning to digital with the following system: a Microlumina slow scan, high-resolution CCD camera connected to a Mac G3, on which we'll run Photoshop and aquire image from scans of negatives placed on a special ($) high illumination output light box. The system is also capable of scanning regular 2x2 and 120 slides. The camera will also be used on my Olympus for brightfield work (not enough light to the camera to permit fluorescence). We're looking at about 12-15 thousand for the whole system. I too have an H600 and have looked at cooled CCD cameras attached to the 35mm port. As a previous respondent mentioned, they don't, as yet offer the resolution that the Microlumina does and they cost about four times as much. Hence our decision to go with scans of negatives. The versatility of the system is also a plus. It is not in place yet, so I don't know what the downside might be, but I think we'll be satisfied. I also am reluctant to give up using the darkroom, although I have been very happy with what can be accomplished using a digital darkroom, i.e., Photoshop. Dwight
Dwight Beebe Institut de recherche en biologie vegetale Universite de Montreal 4101, rue Sherbrooke est Montreal (Quebec) H1X 2B2 Canada Tel: 514-872-4563 FAX: 514-872-9406
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello } } We are looking for a cell-chamber to make time lapse recordings of cells. } Can someone tell me about experiences made with the FCS2 chamber from } Bioptechs? Since we want to make long time recordings focal stability is } of special interest for us. } If soemone has made expiriences with other systems I also would be } interested. } } thanks } Reinhard } } . . . . . . . . . . . . . . . . . . . } Dr. Reinhard Windoffer Fon: (00)49 (0)6131/39 3720 } Universitaet Mainz Fax: (00)49 (0)6131/39 4615 } Anatomisches Institut e-mail: windoff-at-mail.uni-mainz.de } Becherweg 13 } D-55099 Mainz } Germany } . . . . . . . . . . . . . . . . . . .
We use the FCS2 for our live cell imaging, and I think it is fantastic. It is very well designed, and very easy to use. We use it pretty much as they recommend, with their pumps and heaters. They have a web site (www.bioptechs.com) that has a lot of useful info about live cell imaging, as well as technical specs of their systems. It is not cheap, but worth it. My one suggestion: get a couple extra of the thermoplastic glass and gaskets. Careless people ruin these easily.
Your focal stability is going to depend a lot on your microscope system. If you are using an immersion lens, the spring-loaded head will have a tendency, over several hours, to push itself slightly out of focus. Gravity will also play a small role in moving your objectives. The chamber itself is very stable. Dan at Bioptechs is very helpful, and can give you much good advice.
rich
-- --- --- --- -- -- -- --- --- --- Richard J. Dudley (rdudley+-at-pitt.edu) Research Specialist V Dept. of Cell Biology and Physiology University of Pittsburgh http://www.cbp.pitt.edu ---} search BIONET archives at http://www.bio.net {---
I have received multiple copies (at least four today) of a message sent on Dec 8 re:salary survey thoughts. I am certain this is no fault of the author but instead due to some computer system somewhere. Have others been receiving multiple copies as well?
Mick Thomas Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
The correct URL for the web page mentioned below is: http://www.numis.nwu.edu/internet/Staff/wharton/channel.html
Wharton
} } Dear Pedro, } There is a useful section on electron scattering factors in the } International Tables for X-Ray Crystallography. I don't know the volume } number, but it's not the one with the space group tables, and the section is } written by John Cowley. } } While you won't find Doyle-Turner type parametrization of the scattering } factors in this section, the factors are tabulated as a function of } scattering angle, so you might be able to generate such parameters using a } fit. } } I have a web page which may be useful for transforming from the } Doyle-Turner scattering factor parameters to equivalent parameters for the } potential. It is: } http://www.numis.nwu.edu/Staff/wharton/channeling.html } } Good luck } Wharton } ++++++++++++++++++++++++++++++++++++++++++++++++++ } Argonne National Laboratory West } P. O. Box 2528 } Idaho Falls, ID 83403 } Tel: (208) 533-7724 } FAX: (208) 533-7863 } } mailto:wharton.sinkler-at-anlw.anl.gov } } ---------- } } From: Joao Pedro Esteves De Araujo {Joao.Pedro.Esteves.de.Araujo-at-cern.ch} } } To: sinkler-at-apollo.numis.nwu.edu } } Subject: Doyle-Turner potential parameters... } } Date: Thu, Dec 10, 1998, 8:27 AM } } } } } Dear Sir } } I hope you can help me. } } I am trying to find the Doyle-turner potential parameters for } } Yttrium (Y). . Unfortunately in their original paper (Relativistic } } Hartree-Fock X-ray and Electron Scattering Factors, P.A. Doyle and P.S. } } Turner, Acta Cryst., A24, 390, 1968. ) these parameters were not } } calculated. } } Do you know some more recent reference where I could find this? } } Have you some suggestion how to find this information? } } I thank you in advance } } Yours Pedro } } } } } } } } } } } } } } } } } } }
We examine cheese routinely at USU. It sound as if the coating process is allowing the cheese to warm up and the serum and/or the fat may be flowing and carmalizing. Look at Food Structure Vol.12 (1993), pp.475-482 for details on our protocol, If unavailable, let me know and I will get you a copy. Bill
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920
-----Original Message----- } From: "csedax-at-alpha.arcride.edu.ar"-at-Sparc5.Microscopy.Com [mailto:"csedax-at-alpha.arcride.edu.ar"-at-Sparc5.Microscopy.Com] Sent: Friday, December 11, 1998 1:12 AM To: MICROSCOPY-at-Sparc5.Microscopy.Com
Dear all,
we're having problems Au coating cheese samples for SEM observations. They ared freezed and then coated. While being coated, after only a couple of minutes, they get caramelized (does this word exists?). It looks like a film of caramel is produced on the surface. Since we have now experience and no idea
about this type of materials we would like to hear any opinion about coating conditions: should we Au coat them at high pressure for a short time, or low pressure and longer time? what makes this caramel appear?
Please, let us now if any of you have done something like this before. Thanks in advance.
Silvia Montoro Centro Regional de Investigacion y Desarrollo Guemes 3450 3000 Santa Fe Argentina csedax-at-arcride.edu.ar
} -- Begin original message -- } } I have an interesting technical problem that I'm hoping the folks on this list can help me figure out. } I have a colleague who is trying to embed dissected renal tubules (20 } } micron diameter x 500 micron length) for immunofluorescence. He would like to be able to "tack down" individual tubules to some kind of substrate, so that he can keep track of which end he's working from, and would like to cross section them (1-4 micron sections) from end-to-end (vs longitudinal sx). We've tried adhering tubules to Thermanox coverslips and making cryotome sections, but the sections of plastic curled up like a scroll. We've tried Aclar film, and plastic embedding using LR Gold and had the same problem. Is there someone out there who has a suggestion? We have 2 concerns, 1) we don't want to heat the specimen above physiologic temps and 2) we want to keep track of orientation so as to have cross sections of the tubules - adherence to some sort of substrate is one possible solution - } } any alternatives? } } } } Thanks for your help. } } Doug } } .................................................................... } } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : } } : Research Specialist, Principal University of Arizona : } } : (office: AHSC 4212A) P.O. Box 245044 : } } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : } } : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): } } :...................................................................: } } http://www.pharmacy.arizona.edu/exp_path.html } } Home of: "Microscopy and Imaging Resources on the WWW"
Doug:
How about placing the tubules inside another tube such as a length of small intestine or aorta? Adding a bit of gelatine might keep more than one tubule from moving around. The enveloping tube could be fixed to make it less flexible if desired. Just a thought.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
We examine cheese routinely at USU. It sound as if the coating process is allowing the cheese to warm up and the serum and/or the fat may be flowing and carmalizing. Look at Food Structure Vol.12 (1993), pp.475-482 for details on our protocol, If unavailable, let me know and I will get you a copy. Bill
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920 -----Original Message----- } From: "csedax-at-alpha.arcride.edu.ar"-at-Sparc5.Microscopy.Com [mailto:"csedax-at-alpha.arcride.edu.ar"-at-Sparc5.Microscopy.Com] Sent: Friday, December 11, 1998 1:12 AM To: MICROSCOPY-at-Sparc5.Microscopy.Com
Dear all,
we're having problems Au coating cheese samples for SEM observations. They ared freezed and then coated. While being coated, after only a couple of minutes, they get caramelized (does this word exists?). It looks like a film of caramel is produced on the surface. Since we have now experience and no idea
about this type of materials we would like to hear any opinion about coating conditions: should we Au coat them at high pressure for a short time, or low pressure and longer time? what makes this caramel appear?
Please, let us now if any of you have done something like this before. Thanks in advance.
Silvia Montoro Centro Regional de Investigacion y Desarrollo Guemes 3450 3000 Santa Fe Argentina csedax-at-arcride.edu.ar
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Reply to: RE: TEM - serial sections Dear Jeanine,
Cutting serial sections is very easy and Elaine Humphrey and Mike Rock = have told you all you need to know. I can only add a couple of simple = suggestions.
Make sure the top and bottom of the block are parallel with each other and = with the knife edge. =
Shaping the sides of the block so that they are perpendicular to the knife = will sometimes help form ribbons. =
Make sure there is an identifying characteristic in the section which can = be used for orientation. This can be a slight nick in the side of the = block or an easily identifiable part of the sample.
If possible, make the height o fthe block as small as possible. This will = allow you to produce large numbers of sections before having to pick them = up.
Mikes idea for using the sticky tape glue is good but as he points out, = the glue is a little too effective in sticking the sections together. We = apply a product called "Tacky Wax" to the bottom (and/or top) of the block = and find it sticks the sections together but not too securely. As I do = not know the supplier of this product I have been experimenting with = substitutes. The best so far is the "Post-it" glue that is supplied on a = stick from office suppliers. =
Regards,
Paul Webster
Jeannine Caesar wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/apw.htm
id sma004522; Fri Dec 11 16:51:41 1998 Received: from sv-gw1.stsv.seagate.com (sv-gw1.stsv.seagate.com [134.204.14.95]) by auth1.seagate.com with SMTP id IAA06230 for {microscopy-at-sparc5.microscopy.com} ; Fri, 11 Dec 1998 08:51:32 -0800 Received: by sv-gw1.stsv.seagate.com(Lotus SMTP MTA Internal build v4.6.2 (651.2 6-10-1998)) id 882566D7.005C9023 ; Fri, 11 Dec 1998 08:51:02 -0800 X-Lotus-FromDomain: SEAGATE-at-INTERNET To: microscopy-at-Sparc5.Microscopy.Com Message-ID: {882566D7.005C8EB3.00-at-sv-gw1.stsv.seagate.com}
Sarah:
Although you have already received plenty of responses to your question on "digitizing the TEM," it may be interesting to you and others concerned with this issue to look at the article by Alwyn Eades in the current "Microscopy Today", and articles by J. Brink ("Micron" Vol 27 (2), 1997, 129-139), and Sherman and Chiu ("Journal of Microscopy" Vol 188, 1997, 285-289) which deal with resolution of CCD cameras.
I am installing a TEM, and fortunately was able to get a darkroom along with it. For the most part the CCD camera will do very well, with the added advantage of transmitting images to the requester through the local network. Measurements on digitized images obtained directly from the CCD camera or from scanned negatives are conveniently done on the computer monitor, and the whole process is dry and clean. For the highest resolution work I expect to use negatives, and possibly scanned prints of those negatives.
Augusto A. Morrone, Ph.D. Senior Advisory Quality Engineer Analysis Lab Seagate Technology 7801 Computer Ave South Bloomington, MN 55435-5489 Phone: (612) 844-5838 Fax: (612) 844-8247
I have been asked to compare the relative density of capillary beds in different regions of cortex (rodent). My current thinking is to use cardiac perfusion (saline/formaldehyde) followed by perfusion with a dye (which to use?) then cut either vibrotome or frozen sections. If the dye does not leak beyond the bed, quantitative image analysis could be used to determine %difference of dyed vs. undyed cortex. Has anyone done this type of study? Any comments and/or suggestions are welcome.
Thanks,
Richard Mount Auditory Science Laboratory Hospital for Sick Children Toronto, Ontario CANADA
Applied Optical Microscopy will be offered for the 15th year just prior to PITTCON '99, in Orlando, Fl.
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b) a full day on polarized light analysis (both quantitative and qualitative)
c) the opportunity to bring your own samples and use them during the course to develop practical strategies for solving microscopical problems.
For further details and registration information, visit our website { {http://www.MME-Microscopy.com/education} or call me here at the Microscopy/Microscopy office.
{bigger} {bigger} MME {/bigger} {/bigger} is America's first national consortium dedicated to customized on-site training in all areas of microscopy, sample preparation, and image analysis {bigger} .
I need some advice in vacuum impregnation of fractures. We plan to prepare cross sections for SEM analysis from partially fractured aluminum samples (that is a sample which has a fine crack going though its center). This analysis will be followed by opening the crack for an analysis of the fracture surface itself. It is important to protect the crack crevice from the attack by the solutions (water, etchants) used to prepare the cross sections.
In our first attempt, we vacuum impregnated the crack with stop-off-lacquer before handling. After opening the crack, fine particles of the lacquer adhered to the rough fracture surface and we were not able to remove these particles without attacking the fracture surface as well.
Are there any waxes out there suitable for this application?
Many thanks Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
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This is exactly what we did for similar reasons.
We are very happy with the results. It is amazing the "prints" you can obtain using an Epson stylus 800 printer with photo quality paper and Photoshop. Most of our students use this system exclusively now. Some of us "old timers" still like the darkroom at times.
Our system is PC based, but feel free to contact me if you need.
Carol } } } Good morning: } We are also transitioning to digital with the following system: a } Microlumina slow scan, high-resolution CCD camera connected to a Mac G3, on } which we'll run Photoshop and aquire image from scans of negatives placed } on a special ($) high illumination output light box. The system is also } capable of scanning regular 2x2 and 120 slides. The camera will also be } used on my Olympus for brightfield work (not enough light to the camera to } permit fluorescence). We're looking at about 12-15 thousand for the whole } system. } I too have an H600 and have looked at cooled CCD cameras attached } to the 35mm port. As a previous respondent mentioned, they don't, as yet } offer the resolution that the Microlumina does and they cost about four } times as much. Hence our decision to go with scans of negatives. The } versatility of the system is also a plus. It is not in place yet, so I } don't know what the downside might be, but I think we'll be satisfied. I } also am reluctant to give up using the darkroom, although I have been very } happy with what can be accomplished using a digital darkroom, i.e., } Photoshop. } Dwight } } Dwight Beebe } Institut de recherche en biologie vegetale } Universite de Montreal } 4101, rue Sherbrooke est } Montreal (Quebec) H1X 2B2 Canada } Tel: 514-872-4563 } FAX: 514-872-9406
Carol M. Garland, Member of the Professional Staff MC138-78 California Institute of Technology Pasadena, CA 91125
} we're having problems Au coating cheese samples for SEM } observations. } They are frozen and then coated. While being coated, after only a couple of } minutes, they get caramelized (does this word exists?). It looks like a } film of } caramel is produced on the surface. Since we have no experience and no idea } about this type of material we would like to hear any opinion about coating } conditions: } should we Au coat them at high pressure for a short time, or low pressure and } longer time? what makes this caramel appear? } } Please, let us know if any of you have done something like this before. Thanks } in advance. } } Silvia Montoro } Centro Regional de Investigacion y Desarrollo } Guemes 3450 } 3000 Santa Fe } Argentina } csedax-at-arcride.edu.ar
Sylvia -
You'll find the FOODS UNDER THE MICROSCOPE Web page http://www.cyberus.ca/~scimat/ quite helpful. Contact its author (who doesn't read this listserver) if you have further questions: Milos Kalab {kalabm-at-EM.AGR.CA}
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
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All,
All the ergonomic chairs we can find contain (magnetic?) metal parts. If they are moved at the microscope, their motion affects the position of the zero-loss peak of a GIF. The effect can be rather annoying. We really need ergonomic but non magnetic chairs made from plastic or wood. Does anyone out there know of a supplier?
Dr. Christian Kisielowski
National Center for Electron Microscopy Lawrence Berkeley National Laboratory One Cyclotron Road, Bldg. 72/125 Berkeley, CA 94720 USA
Tel: (510) 486 4716 Fax: (510) 486 5888
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begin: vcard fn: Michael O'Keefe n: O'Keefe;Michael org: {center} {img src=http://mercproj.lbl.gov/logo.gif} {br} {b} {big} National Center for Electron Microscopy {/big} {/b} {/center} email;internet: maokeefe-at-lbl.gov title: Deputy Head x-mozilla-cpt: ;0 x-mozilla-html: TRUE version: 2.1 end: vcard
Hello, Our company produced verry large chamber for SEM. You are invited to visi= t our site web: http://perso.wanadoo.fr/opea.hoan/ Our realizations for: CEAT (Centre d'Essais Aeronautique de Toulouse - FRANCE) OCE (Holland) Hoan OPEA 114, rue de la Jarry 94300-VINCENNES (FRANCE) Tel.33.1.4328 3496 Fax:33.1.4328.0364
Marienhoff a =E9crit:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } } Dear all, } } We want to integrate two separate motorized tables of five axis each in= to our } large chamber SEM. Who knows suppliers of these tables? } } Best regards } } Peter Marienhoff } ___________________________ } Dr. Peter Marienhoff } VisiTec Microtechnik GmbH } Karl-Marx-Str. 14 } D-23936 Grevesmuehlen } Germany } } Fon: +49-3881-790-47 } Fax: +49-3881-790-48 } email: pmarienhoff-at-visitec-em.de } http://www.visitec-em.de
Two groups here at the MBL have been using the Delta TC3 system from Bioptechs. It has been working very well for our purposes but the FCS2 would probably be better for time lapse recordings. Dan at Bioptechs has been of much help along with their web page: www.bioptechs.com.
Thanks, Louie
At 9:40 AM +0100 12/11/98, Reinhard Windoffer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
www.dvcco.com No cooling needed, color integration to 10 sec $4995 !!! These are 10 bit cameras.... 60dB signal to noise not 50. See model info below This is an informational update on a new type digital camera on the market from the manufacturer. This is current info, not some old, me too camera info rehash from years past by someone who paid too much. CURRENT SHOW: CELL BIOLOGY / SAN FRANCISCO MOSCONE BOOTH 15 12/13 - 12/16/ 1998 Going on as you read this! Web site: www.dvcco.com this is the most current site for you to bookmark ****** Please look at image gallery / color images/ biomedical images HERE IS WHAT WE CAN NOW OFFER THAT THE OTHERS CANNOT FOCUSING ON THE MICROSCOPY MARKET. * A 12fps focusable image finally.... No 60% signal loss via a noisy RGB sequential filter and slow response of 1 frame every 4 seconds etc. -at- $8k+. * Full RS232 Control outboard of the camera for 10 second integration with no noise or up to 30dB gain at 12fps so you can have it either way. Remember if you start with a 60dB signal and add gain you go from 10 bits down to 8 but if you start with a competitors camera at 50dB which is 8 bits only marginally you go down to 7 bits or 6. So with 50dB your are starting out with 7.5 to marginal 8 bits before you even add gain. Who wants less than 8 bits in the first place??? Your camera guy might say Oh we use that sensor also BUT....what he doesn't know is that due to his manufactures rush to get the camera to market they did a sloppy auto route job of the board and ended up with 50dB signal to noise Vs our 60dB. It is how much of the signal to noise of the sensor YOU PRESERVE that counts in a design. Yes, tell your camera/microscope salesmen they can stop telling everyone they need cooling just because of a little integration in the range of 0-20 seconds. NOT SO if they have YOUR best interest in mind. This was the big surprise at the Neuroscience show / virtually everyone automatically parroted cooling in the same breath with integration.... * Color integration.... one snap and its done, no overlays. * We offer the whole system digital camera, cable, supply, and frame grabber PCI/PC by Epix / PIXCI-D for about $6300!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! $4995+200+150+995. * Yes we have a new Mac interface also / ITI board, but please if you have a choice go PC/ PCI. So what if NIH Image is free, so is this software.... We can even offer you the computer 400Mhz+ in a system package. * We are a US camera manufacturer, with full control of our design and OEM engineering capability. OEM quantity orders are no problem. * Yes it is TWAIN compatible and the Epix board for $995 even has all the free software you will ever need to grab and evaluate your image via some of the other fine programs that are out there along with custom color balance software for the DVC 1300C * The DVC1300 Mono or DVC1300C / RGB camera can BOTH be USED on the same RS422 digital only Mono Epix frame grabber. You do not need a special frame grabber for color and the display of the RGB image is on your computer monitor...not some expensive RGB monitor. Were talking digital here, not analog so we have more avenues and flexibility than with the old analog cameras. * When evaluating cameras do it via S/N - signal to noise first....at 12 frames/sec We offer 10 bits, no more... Even with integration. Be careful, others will say they do 16 bits or 14 etc.... but at how much integration time... * Yes we have cooling to -40 if needed. Most integration out to perhaps 10 seconds or a little more DO NOT. Cooling is an expensive add on that also could reduce the life of the camera if the seal leaks later..... * The sensor is effective a grade one sensor. Before you purchase a camera ask what grade sensor you are getting. A well known mfgr ships grade 3 sensors and charges you 10K+ for the privilege. Models: DVC1300 Monochrome 1300 x 1030 pixels-at- 12fps or 1300 x 515 -at- 24fps !!!!!!!! DVC1300C Color RGB 1300 x 1030 pixels -at- 12fps
* Here is an idea: For the price these systems are offered you can outfit your lab with 2 if not more cameras with one frame grabber and 1 each of the RGB and mono DVC cameras instead of some pricey mono camera with a noisy lossey sequential filter and no frame grabber for the price, or some built in one with no flexibility. We fit a niche market...Most people that would like a good fluorescent image or other with decent signal to noise for 10 bit or less with gain picture and all the gain 30db or integration they need. Yes, if you go past 20 or 30 seconds then you will need cooling. The explanation of the color filter/ Bayer pattern is on the web site under DVC1300C. The Bayer pattern reduces aliasing also. Every wonder why at the Vision or Photonics East/West shows you see these color camera manufacturers showing big.......colorful teddy bear's etc. Its because they do not have a decent Megapixel color camera, only low resolution analog that is 7 bits or 48 dbS/N. Big and smooth does not show you the detail. There is a complete FAQ ....Frequently asked questions section in the index for your reading pleasure. We hope this helps let you know there is some radically new cost effective technology out there that you should be aware of. Double click on the images to blow them up for your review and download. Thank you for your time and if I offended anyone, God help me, I have seen enough boring messages that I know this does not fall into. Regards, Rich
Richard Klotsche DVC Company Sales Manager 619-444-8300 619-444-8321-fax dvcco-at-aol.com www.dvcco.com
In the past I was deeply involved in the design of SEM cryo systems and t= he problem you relate is typical of what I would term "old fashioned" sputte= r coating systems, too hot!.
Sputter coating is one of the biggest problem areas in any form of SEM, i= t often cooks and very often decorates a specimen.
May I suggest you use the following procedure to look at the specimen UNCOATED, then you will find the source of your problem.
1. Run at 2kV =
2. Depending upon the microscope you have you will probably need abo= ut 50uA emission =
3. Use a spot size SMALLER than you would normally use as this is a safety device 4. Run at a maximum of 15mm WD
Always look at a specimen in this way first and then you will know what t= he real specimen looks like. You will be surprised just how many specimens=
will be usable without coating. =
If you do decide to coat do so at the lowest voltage that your coater wil= l provide (probably about 800v) and run for no longer than 1 minute at 20mA= . =
To improve the coating efficiency move the sample along the cold block, backwards and forwards, as you coat.
For more help please feel free to contact me with details of your cryo system and your microscope.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
The electron source sets the limits of the SEM system. All that our condenser lenses do is to reduce the size of the source by throwing electrons away. A crude rule of thumb is the resolution attainable will = be 10,000X smaller than the source size (50 micron source 50 Angstrom resolution). The answer to the FEGSEM sucess is its source. About 1000=
times brighter than a W hairpin and measured in nm rather than micron there are far more electrons available to us per unit area. This makes = it possible to form smaller probes with sufficient current to gereate a high=
level of signal. The FEG really comes into its own at low voltage for a number of reasons.
1. The W hairpin system loses efficiency as you move away from the design kV. The gun has to be designed with a certain minimum anode to cathode distance (about 1mm for every 2kV) in order to prevent discharge = at the highest kV. As soon as you move away from the higest kV the system i= s no longer optimised and the gun becomes less efficient. You can improve the gun performance by raising the anode and moving the filament forward.= =
Even with these modifications the low kV performance still falls short because of the increased level of aberrations in the system. Lower operating lens currents and accelerating voltages mean that small irregularities have a greater effect. Add to these problems the increase= d beam spread when it strikes the specimen at lower voltages and you see w= hy the resulting performance is pretty poor.
2. Using a FEG we do not have the problems with gun design and the source is so much improved that even when using small spot sizes at the specimen (compensating for beam spread in the specimen and that due to aberrations) there is more than enough current to generate a good signal.=
Take a W hairpin system at 2kV and you will be doing well to obtain 20,00= 0X with any quality, go to a good FEG SEM and 90,000X at 2kV is not such a problem. The best FEG systems from my experience are the ones using a double detecting system, here the choice of signals makes the operation o= f the instrument and an understanding of imaging information an operator's dream, we can select the signal that we want.
Dual detector imaging often known as semi in lens imaging, relies upon th= e SE being attracted to a detector which is situated above the final lens pole piece. Initially commercial instruments with two ET detectors used the field from the final lens to "pull" SE into the upper detector. ISI were first with their SS Series and then Hitachi with the S570 took this route. In these instruments they allowed the specimen to be anywhere between 100% out of lens, through to what we would consider to be a WD of=
-5mm. That is 5mm up inside the pole piece! Problems arose if you were using magnetic materials, hence the development of a twin detector system=
by Hitachi which used a field coil to drag electrons up to the detector a= nd a pole piece design which retained the lens field within the pole piece. =
The twin detector system gives the microscopist the best of all worlds.
1. The upper detector provides an opportunity to sift out the BSE an= d obtain a pretty pure SE image. The high lens strength at very short WD (~3mm) enables the instruments to reach very high resolution levels. The down side of this is, as SE are effected by charge, this mode is more pro= ne to charge problems. However at {5kV in my experience we have very few problems provided the operator knows about low damage techniques.
2. The lower detector offers the type of contrast which we all see i= n our conventional single detector SEM, SE+BSE. The up side is that the BS= E contribution to this image results in far fewer charge problems and may b= e a good compromise for the biologist.
3. Add a BSE detector to such a system, and you can move in any direction "pure" SE or SE + BSE or pure BSE. Drop the kV and the BSE becomes even more interesting as the volumes of material involved almost mimic SE volumes. Not to be discounted for biological applications.
So there we are I hope this helps those who have no experience with the new era, that of dual detector FEG SEM.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
THANK-YOU to all who responded to my questions. I received many informative and wonderful responses. I am currently writing a submission to suggest we keep at least half our darkroom for the developement of negatives and for printing the occasional negative BUT I am also suggesting that we purchase a good quality scanner with associatied software and computer in compensation for losing part of our darkroom. Digitizing the TEM appears to be a very expensive alternative but I'll also furnish the 'Powerful Ones' with information on this as well. I will post a summary of the information I received from all the wonderful Micrscopy Listserver people as soon as practical. Again,
Thank-you!!!
Sarah Ellis
Trescowthick Research Centre Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne 8006 Victoria Australia
Silvia, Are you trying to keep your sample frozen or even cooled during the coating +imaging? I have a couple of thoughts depending on the equipment you have--- 1. if you are using a cryo-SEM, keep the sample small enough to fit into a 2 mm rivet. Place a second rivet onto the surface, freeze in LN slush, transfer to the specimen preparation chamber and fracture at -196C, transfer the sample onto the SEM stage, heat the stage ( -100C) and allow the surface ice to sublime (etch) for 4 min. Try imaging your sample "uncoated" at 2.5 or 5 kV. If you are unhappy with the image, transfer the "etched" sample back to the table in the prep chamber and coat 30 sec. with Au. This worked on "soft serve froz. yogurt" and some protein gels 2. if you are doing all of this at ambient temperatures, you might be able to cool the table inside the sputter- coater by pumping ice water through it. We managed to do this with an an old ISI PS-2 coater.In this case I would increase the distance from the target to your sample as much as is possible. In that same coater the target housing can be pulled up and the table adjusted as well. 3. if you can use your sputter coater to evaporate C or find a vacuum evaporator, you can maximize the distance from the C thread or rods as you evap. for 1 or 2 sec at 1 sec intervals and again try to use a heat sink such as a copper block in liquid N 4. lastly, you might try some form of fixation: --freeze-subsituting the sample in acetone +osmium at -80C. If you do this last procedure--be prepared to follow up with a dehydration series and critical point drying. The sample should be more conductive...but I would still try to coat it with Au or C. --follow a procdure used by M. Kalab (I have to check the reference) but I believe a series of 3mm dia. holes were drilled into an aluminum stub, the "soft" dairy sample was placed into each hole and covered with low melting pt. agar. The entire stub plus samples were fixed at ambient T with gluteraldehyde followed by osmium tetroxide, ethanol dehydration and critical point drying. Before the samples were sputter-coated,the surface of each was touched with the sticky tape on a second aluminum stub and both were then sputter- coated. I recently tried this with whey protein gels but only used vapor fixation with osmium before dehydrating and coating. I'll send references asap. Let me know what you found to work. Rosemary
Can anybody help me? I am in urgent need of SEM photos, good drawings and/or colour plates of the following genera or species: Dermatophagoides pteronyssinus; facies and whole body Larva and adult of Neotrombicula autumnalis; dorsal view Tetranychus sp.; facies and dorsal view - and if possible, drawings or photos of the stages in their mating ritual + photo of infested plant Varroa jacobsoni; facies, dorsal and ventral view + photo of infested bees Scheloribates laevigatus as intermediate host of the tapeworm Moniezia expansa + drawings of lifecycle of the tapeworm Dirocheles phalaenodectes; photo + drawins of lifecycle in the tympanal organ of a noctuid moth Demodex folliculorum or D. brevis; single specimen + drawing of several specimens in hair follicle Sarcoptes scabei; dorsal and ventral view + drawing of lifecycle in the human skin Dorsal view of male and female Ixodes ricinus, and colour plate of engorged animal. Also SEM photos of the chelicera. Arrenurus conicus; male and female, dorsal view Glycyphagus canestrinii; male and female, dorsal view Pergamassus crassipes, male and female, ventral view Analges clavipes, male and female, ventral view Labidocarpus megalonyx, Geckobia loricata, Caelculus echinipes, Carabodes elongatus, and Pelops phaenotus, all dorsal views Tegeocranus sp.(dorsal view) and Belba sp. (lateral view) with nymphal exuvial remains intact Pelops acromius larva, tritonymph and adult, all dorsal views
The pictures are to be used in an exhibition at the Bergen Aquarium, and we need them before January 5th. And they must be high quality.
If you can help me, please send me an e-mail (eli.amundsen-at-zoo.uib.no) naming the pictures you can supply, and your price of delivery and I will get back to you with details of payments and postal adress.
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A very easy way to evaluate capillary beds is to use the presence of endogenous peroxide in the red blood cells present in the capillaries. Remove the tissue from the animal and immediately cool...do not wash out the blood but immersion fix in 4% paraformaldehyde. Then simply react vibratomed tissue sections with DAB and the resultant dark brown reaction product makes it quite easy to trace the capillaries by light microscopy. The reaction can be further enhanced by exposing the sections to peroxidase-congugated IgG to increase the background staining of the blood plasma.
See the reference: Sherman, D. and W. Paull. (1985) "New Method for Visualization of Vascular Networks in Nonperfused Fixed Tissues". Stain Technology 60:2 p.89.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Suggestions needed!
I have been asked to compare the relative density of capillary beds in different regions of cortex (rodent). My current thinking is to use cardiac perfusion (saline/formaldehyde) followed by perfusion with a dye (which to use?) then cut either vibrotome or frozen sections. If the dye does not leak beyond the bed, quantitative image analysis could be used to determine %difference of dyed vs. undyed cortex. Has anyone done this type of study? Any comments and/or suggestions are welcome.
Thanks,
Richard Mount Auditory Science Laboratory Hospital for Sick Children Toronto, Ontario CANADA
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If chemistry associated with cracking is of concern (sounds that way), then almost anything added to the crack will be problematic. Either it will add it's own chemistry, or you may clean off offending material along with the impregnating material. If it is at all possible, I would suggest sectioning
(careful, dry cutting) the crack into two pieces, one for x-sect work and the other to open.
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I need some advice in vacuum impregnation of fractures. We plan to prepare cross sections for SEM analysis from partially fractured aluminum samples (that is a sample which has a fine crack going though its center). This analysis will be followed by opening the crack for an analysis of the fracture surface itself. It is important to protect the crack crevice from the attack by the solutions (water, etchants) used to prepare the cross sections.
In our first attempt, we vacuum impregnated the crack with stop-off-lacquer before handling. After opening the crack, fine particles of the lacquer adhered to the rough fracture surface and we were not able to remove these particles without attacking the fracture surface as well.
Are there any waxes out there suitable for this application?
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-- ************************************ Maite Caldes-Rouillon Institut des Matériaux de Nantes 2 rue de la Houssiničre, B.P. 32222 44322 Nantes Cedex 3 http://www.cnrs-imn.fr Tél: +(33) 02 40 37 64 17 Fax +(33) 02 40 37 64 18 ************************************
Since you are doing relative densities, and don't need absolute measures such as total cross-sectional area how about this: do corrosion-casting of the brains--I don't have the references handy, other than Fred Hossler's article in the Sept. '98 Microscopy Today. (Caveat--I'm tech. editor.)
Basically, methacrylate resin is injected into the blood system, then the organ of interest is removed after the resin has polymerized. The brain in your case. Then remove the areas of interest from the brains and digest away the tissue. If you've removed equal sized cubes (etc.) from the different areas/treatments, and if you've completely filled the capillaries, etc., then the different weights of the casts would directly correlate with the relative differences in capillary densities. Volume of the capillaries could be determined by immersion in water--the small amount of displace being the major headache. Alternatively, given the weight and the known density of the resin, volume could be calculated.
The obvious problem is cutting away the resin casts of the capillary beds from the casts of the larger vessels. This is possible, if mind-numbingly delicate work.
Phil
} Suggestions needed! } } I have been asked to compare the relative density of capillary beds in } different regions of cortex (rodent). My current thinking is to use } cardiac perfusion (saline/formaldehyde) followed by perfusion with a dye } (which to use?) then cut either vibrotome or frozen sections. If the dye } does not leak beyond the bed, quantitative image analysis could be used } to determine %difference of dyed vs. undyed cortex. } Has anyone done this type of study? Any comments and/or suggestions are } welcome. } } Thanks, } } Richard Mount } Auditory Science Laboratory } Hospital for Sick Children } Toronto, Ontario CANADA
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 oshel-at-terracom.net
Our company, ElectroImage, Inc. produces and markets the TEMSCAN System = to digitize TEM negatives. =20
Whenever there is a thread about the various methods to obtain digital = images from a TEM, our TEMSCAN system is generally mentioned by one of = our customers as an alternative. What is not mentioned is what = separates this system from flatbed scanners that have a transparency = adapter. =20
The TEMSCAN System is composed of a MicroLumina digital camera, = high-frequency light box, and copy stand. The result of this system is = a 9MB grayscale file that is 2700 x 3400 pixels. Scan times are less = than a minute for typical density TEM and SEM negatives. If resolution = isn't as important, the scan times can be decreased by scanning with = less resolution. Although many flatbed scanners are able to scan = transparencies, the key element that sets the TEMSCAN system apart is = the lens. With a lens attached to a scanning camera, cropping into the = negative doesn't result in a loss of resolution. For example, if the = area you wanted to scan on the negative was only one-fourth of the whole = image, and both the scanner and the camera were capable of the same = resolution (1000 x 2000 for example), the flatbed scanner would only = give a 500 x 1000 pixel image while the camera can simply focus = closer and still produce the full 1000 x 2000 resolution. Since most = flatbed scanners are much larger than 4 x 5 inches (typically 8 x 10 = or larger) you automatically throw away two thirds or the total = potential resolution of the device. =20
The MicroLumina can also be used for brightfield microscope work = producing a file of 26.1MB.
I would welcome the opportunity to scan in any negatives you wish to = send me. I will return your negatives and give you a CD with your = scanned images and hard copy to show the quality of this system.
Matt Irwin=20
ElectroImage, Inc. 277 Northern Blvd. Suite 101 Great Neck, NY 11021 Phone: 516-773-4305 Fax: 516-773-2955 E-mail: sales-at-electroimage.com Website: http://www.electroimage.com
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} To all concerned: {BR} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Our company, ElectroImage, Inc. = produces and=20 markets the TEMSCAN System to digitize TEM negatives. = {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Whenever there is a thread about the = various=20 methods to obtain digital images from a TEM, our TEMSCAN system is =
generally mentioned by one of our customers as an alternative. = What is not=20 mentioned is what separates this system from flatbed scanners that have = a=20 transparency adapter. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} The TEMSCAN System is composed of a = MicroLumina=20 digital camera, high-frequency light box, and copy stand. The = result of=20 this system is a 9MB grayscale file that is 2700 x 3400 pixels. = Scan times=20 are less than a minute for typical density TEM and SEM negatives. = If=20 resolution isn't as important, the scan times can be decreased by = scanning with=20 less resolution. Although many flatbed scanners are able to scan=20 transparencies, the key element that sets the TEMSCAN system apart is = the=20 lens. With a lens attached to a scanning camera, cropping into the =
negative doesn't result in a loss of resolution. For example, if = the area=20 you wanted to scan on the negative was only one-fourth of the whole = image, and=20 both the scanner and the camera were capable of the same resolution = (1000 x 2000=20 for example), the flatbed scanner would only give a 500 x 1000 pixel = image while=20 the camera can simply focus closer and still produce = the full=20 1000 x 2000 resolution. Since most flatbed scanners are much = larger=20 than 4 x 5 inches (typically 8 x 10 or larger) = you=20 automatically throw away two thirds or the total potential resolution of = the=20 device. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} The MicroLumina can also be used for = brightfield=20 microscope work producing a file of 26.1MB. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} I would welcome the opportunity to = scan in any=20 negatives you wish to send me. I will return your negatives and give you = a CD=20 with your scanned images and hard copy to show the quality of this=20 system. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Matt Irwin {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D2} ElectroImage, Inc. {/FONT} {/DIV} {DIV} {FONT size=3D2} 277 Northern Blvd. {/FONT} {/DIV} {DIV} {FONT size=3D2} Suite 101 {/FONT} {/DIV} {DIV} {FONT size=3D2} Great Neck, NY 11021 {/FONT} {/DIV} {DIV} {FONT size=3D2} Phone: =20 516-773-4305 {/FONT} {/DIV} {DIV} {FONT=20 size=3D2} Fax: = =20 516-773-2955 {/FONT} {/DIV} {DIV} {FONT size=3D2} E-mail: {A =
As most of the answers to my queries on digitization of the TEM were posted to the Microscopy Listserver, you may feel that this summary is not needed. However, due to the large response generated, I feel the topic is worth summarising. Please note that this summary is my own interpretation of the overall views expressed and may not therefore be entirely accurate. Most of the responses received were related to imaging for biologicaI sciences. I have no affiliation with any commercial company. It appears that the side mounted CCD cameras are the most popular mainly due to the very high cost of the larger CCD cameras which are fitted below the screen. These latter cameras also take up valuable knee space but if you have lots of $$$ then it appears that the camera of choice here is a 2K x 2k CCD camera. TV rate cameras or video cameras are really not suitable as the images generated are of poor quality. The general consensus is that the camera of choice in todays market is a 1024 x 1024 CCD camera with a dynamic range of at least 12 bit and a frame rate of 10 or more at full resolution. The cost of such a camera appears to be in the vicinity of US$50K -US$70K. The company 'Gatan' was frequently mentioned as 'good'. Other companys mentioned were 'Soft Imaging System Corp.' and 'Dage'. The company of choice will advise on whether you need any other accessories depending on your needs and I did not receive enough information on this to make a summary. Software for these cameras is generally provided by the camera manufacturer and it is wise to ensure that the software is not a proprietary image format and is compatible with other software in use. Having said all that, I found the overwhelming response from nearly every reply was that the 'old fashioned' negatives were still superior in terms of resolution compared with the digital 'equivalent'. The respondents that were lucky enough to have a CCD camera attached to their TEM generally still relied on film for publication quality prints and used the digital images for 'working' prints. In conclusion, unless you have lots of dollars, the way to go appears to be to scan your TEM negatives using a good quality film scanner and manipulate them with the associated software (Adobe photoshop). These scanned images have the advantage that they can be easily cropped, annotated and assembled into plates and can produce publication quality prints. In addition, negatives are still the most archival, platform independent and cheapest form of data storage.
I hope this summary is of some use,
Cheers
Sarah Ellis
Trescowthick Research Centre Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne 8006 Victoria Australia
Attention listservers: One of my government customers has decided to part with their ETEC = SEM. They have asked me to find it a good home before it goes out on = surplus. The recipient MUST be associated with the US government. The = instrument is in excellent condition and has been under full service = contract for at least 20(yes, that's TWENTY) years. It is has been = upgraded with the following options:
Computerized mag control with text writer Digital Scan Generator DIGISEM SEM Digital Imaging package Dapple(Noran) PC based EDS Analyzer(KEVEX Be detector needs to be = rebuilt) GW BSE NEW 8 CFM inline roughing pump Fairly new NESLAB water recirculator/chillier
For those of you who are not familiar with the ETEC SEM's, it's standard = features are:
2.5 to 30KV - Tungsten filament Smallish specimen chamber Five axis eucentric specimen stage - Tilts to +45 degrees Twin viewing CRT's 2000 line Photo CRT Fast pump down time - approx. one minute from atmosphere to beam turn on
Remember, you have to be associated with the US gov't to get this = instrument.
Please email me at: gary.easton-at-scannerscorp.com
Gary M. Easton, Pres. Scanners Corporation 1-800-466-SCAN Third Party SEM Service EDS and Digital Imaging upgrades
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} Attention listservers: {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} One of my government = customers has=20 decided to part with their ETEC SEM. They have asked me to find it = a good=20 home before it goes out on surplus. The recipient=20 {STRONG} {EM} {U} MUST {/U} {/EM} {/STRONG} be associated with the US=20 government. The instrument is in excellent condition and has been = under=20 full service contract for at least 20(yes, that's TWENTY) years. = It is has=20 been upgraded with the following options: {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Computerized mag control with text=20 writer {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Digital Scan Generator {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} DIGISEM SEM Digital Imaging = package {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Dapple(Noran) PC based EDS = Analyzer(KEVEX Be=20 detector needs to be rebuilt) {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} GW BSE {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} NEW 8 CFM inline roughing = pump {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Fairly new NESLAB water=20 recirculator/chillier {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} For those of you who are not familiar with the ETEC = SEM's,=20 it's standard features are: {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} 2.5 to 30KV - Tungsten filament {/FONT} {/DIV} {DIV} {FONT size=3D2} Smallish specimen chamber {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Five axis eucentric specimen stage - = Tilts to=20 +45 degrees {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Twin viewing CRT's {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} 2000 line Photo CRT {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Fast pump down time - approx. one = minute from=20 atmosphere to beam turn on {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {FONT=20 color=3D#000000} {STRONG} {EM} {U} Remember {/FONT} , you have to be = associated with the=20 US gov't to get this instrument. {/FONT} {/U} {/EM} {/STRONG} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Please email me at: {A=20 href=3D"mailto:gary.easton-at-scannerscorp.com"} gary.easton-at-scannerscorp.com= {/A} {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Gary M. Easton, Pres. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {FONT size=3D2} Scanners=20 Corporation {/FONT} {/DIV} {DIV} {FONT size=3D2} 1-800-466-SCAN {/FONT} {/DIV} {DIV} {FONT size=3D2} Third Party SEM Service {/FONT} {/DIV} {DIV} {FONT size=3D2} EDS and Digital Imaging upgrades {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {/BODY} {/HTML}
Need some help here. We are new to the cryo side of ultramicrotomy. I would be interested in the clinical research applications of cryoultramicrotomy. We wish to do immunocytochemistry on cultured myoblasts and myotubes; all the antibodies that we wish to examine do not withstand fixation, and we are assuming that we need to approach this from a cryo point of view. There have been a couple of reports on their localization using freeze-fracturing (and I know even less about that). Before we start getting info and quotes from manufacturers, I'd like some more information about this and other possible applications. Would someone be able to give me a ball-park figure of how much it would cost to perform cryoultramicrotomy taking into account technical time, and consumables (minus the cost of primary antibody)? Thanks in advance for any advice Ronnie Houston Texas Scottish Rite Hospital for Children Dallas, TX 75219
Yes no problem. That is of course if the gun was an oil filled type. If it is the older = epoxy type you will have to first get rid of the epoxy and clean it up = before using some other HV epoxy to fill it again. Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
-----Original Message----- } From: "Corvos-at-aol.com"-at-sparc5.microscopy.com = [SMTP:"Corvos-at-aol.com"-at-sparc5.microscopy.com] Sent: Tuesday, December 15, 1998 3:18 AM To: Microscopy-at-sparc5.microscopy.com
LUC,
Can you confirm again that Santovac 5 is OK to use on the gun... =20
I received an inquiry asking what would be a most suitable stain for student's to perform a quick blood stain to differentiate lymphocytes, macrophages, neutrophils etc (basically a DNA vs cytoplasm stain). My background is mostly in EM and so I have trouble thinking past Toluidine Blue O, differentiated with acid alcohol. That would do, but I expect there is a better alternative for smears. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
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Biological SEM users: I got very little response to the message I posted a week ago (see below). It appears that at the present time there is not a great deal of biological data from FEG-SEMs. To help us determine the best instrument to consider for biological data accumulation, I would also like to explore the pros and cons of the FEI/Philips'ESEM with the low vacuum instruments available from other manufacturers. I have the brochures, etc from a number of companies so am not looking for more of the same. I also have looked into the basic design differences and detector differences.
What I would appreciate is comments from users based on their experiences. Why did you choose the instrument you did? What type of information are you getting from your instrument of choice that you could not get from a conventional SEM? Do you also have a cryo unit and x-ray detector and, if so, what are the limitations for their use in low vacuum modes? How many hours per week is your instrument used and what percentage is in the low vacuum mode? Do you have other conventional SEMs or is this your only instrument?
Any other information would be appreciated. If the response is adequate, and off-line, I will try to summerize.
Thanks, Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
----------------------------------------------- Earlier post (12/7/98): I would like to hear from microscopists who have used FEG-SEM's and particularily the semi-in-lens models for biological applications. I would also appreciate info on recent papers where this instrumentation was critical to the biological research reported. Also any recommendations as to ideal samples for testing these microscopes for biological application? Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
(InterLock SMTP Gateway 3.0 for Microscopy-at-sparc5.microscopy.com); Tue, 15 Dec 1998 12:00:27 -0600 Received: by na2.dow.com (Internal Mail Agent-1); Tue, 15 Dec 1998 12:00:27 -0600 Message-Id: {D209047D0257D21191770000F8C991D916BB15-at-mante35a.nam.dow.com}
Could anyone suggest a source for PtSi2 ? (powder { 200 micron is preferred ). I only need a few grams. I thin film (i.e. similar to the NiOx TEM standard) would be ideal. Yes - I did check with Goodfellows and Aesar. They will provide the material as a special order but the minimum order was higher than I needed. thanks, Steve Rozeveld
Steve Rozeveld The Dow Chemical Company Analytical Sciences Laboratory 1897 Bldg., Door E43 Midland, MI 48667 517-636-5167 office 517-638-6443 fax
We have a Kevex Delta V EDS system available for parts. It is available to anyone willing to pay for the shipping.
We have kept the detector, amplifier, and HV power supply but have replaced the computer section with a new Windows-based system.
The system contains a PDP-11/73 processor with 2-4 MB of memory, 1 10 MB bernoulli drive, a 170 MB SCSI hard drive, and an ethernet card with software. It has the full complement of Kevex software including Quantex, Advanced Imaging, and Automated Image Analysis. It has both RT-11 and TSX operating software.
If you or someone you know is interested, contact me via one of the means below.
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
I have had some success with overlaying oil on a culture chamber in order to prevent evaporation when using a 37=B0 heating plate for open chamber microscopy. So far, I have used only light mineral oil. The problem is that you have to put quite alot of oil on top to cover the surface. I am wondering if anyone has found an oil that spreads better to form a barrier with less volume.
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
This question would be best asked on Histonet--you'd get lots of quick replies from folks who do blood smears and any other histo work for living. I've included the Histonet listserver information below. Including some who teach histotech and write texts on it.
If you can find a copy of "Staining Procedures" (Biological Stain Commission), there are several stains given. Any histo text ought to have these as well. The usual stains last time I did this were Wright's and Giemsa for thin film. If you receipes, let me know--I can send you these and a few others. Mind, mine are older ones.
Phil
WHO RUNS HISTONET? The list is run by Linda R. Margraf, M.D. and Herb K. Hagler, Ph.D. using hardware and software owned by the University of Texas Southwestern Medical School, Department of Pathology in Dallas, Texas. If you have any questions or problems with Histonet please contact Linda Margraf at LMargraf-at-childmed.dallas.tx.us.
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} I received an inquiry asking what would be a most suitable } stain } for student's to perform a quick blood stain to } differentiate } lymphocytes, macrophages, neutrophils etc (basically a DNA } vs cytoplasm } stain). } My background is mostly in EM and so I have trouble } thinking past Toluidine Blue O, differentiated with acid } alcohol. That would do, but I expect there is a better } alternative for smears. } Jim Darley } } ProSciTech Microscopy } PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } Great microscopy catalogue, 500 Links, MSDS, User Notes } ********************** www.proscitech.com.au *****
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net
Email: gorry2+-at-pitt.edu Name: Michael Gorry School: University of Pittsburgh
Question: Can you recommend any books or journal articles about the basics of fluorescent microscopy photography? I know nothing about cameras and am certain that I could improve the quality of the pictures that I have been taking if I knew more about the camera and exposure times, aperature sizing, etc. It is a Nikon microphot-FXA.
I could find no introductory information from the Nikon people.
Any help would be appreciated.
Thanks
Mike
Michael Gorry E1105 BST University of Pittsburgh Arthritis Institute Pittsburgh, PA 15261
In a message dated 98-12-15 16:38:37 EST, gorry2+-at-pitt.edu writes:
{ { Question: Can you recommend any books or journal articles about the basics of fluorescent microscopy photography? } }
Mike,
There is a small book that was produced by Wild Leitz in Germany, entitled "Fluorescence Microscopy: Principles, Instruments, Applications," written by Eberhard Becker. Tips and tricks of photography are discussed on a couple of pages in the book.
Are you having troubles with overexposed red fluorescence? Depending on the vintage of the scope and the photodetector, there may be problems with reduced sensitivity of the detector in the red end of the spectrum. The result: seriously overexposed and washed-out photos if you set the camera for the correct film speed.
You can compensate for this by multiplying the film speed (ASA) by about 4 or adding about 6 to the DIN number when you take the photographs. This and other topics are addressed in the book.
There is a chance that Leica still has some of these books. Call Leica Customer Service at 1-800-248-0123 to inquire.
Hope this helps!
Cheers,
Bob **************************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Research Microscopy Products *****************************************
To Michael Gorry, U of Pitt: re: Fluorescence Photomicrography
Your FXA may have too many "bells and whistles" for you to plow through to understand what's happening with your photography. With our rather simple set up, Nikon Optiphot with Microflex UFX II, I get excellent pictures using Kodak Professional Ektachrome 1600, shot at 1600. Make sure your camera can spot meter, that is to be able to read the brightest part of your field and not average the entire field. This way, you meter the very strongest signal, set that value in the memory of the timer, reposition your slide for the most pleasing shot and then expose. Background should be dark to black...if you have flares or glare, there is probably a diaphram somewhere between the lamphouse and the microscope to stop down (but not to the point of vignetting your picture). Good luck! I have an FXA instruction manual with a photography section if you need a copy. Bob Santoianni Emory University Hospital Atlanta, GA robert_santoianni-at-emory.org
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A great, inexpensive and very readable initial sourse is : Herman, B.(1998) Fluoresence Microscopy.BIOS Scientific Publishers Limited. ISBN: 0-387-91551-6. This is part of the Royal Microscopy Society series published by Springer-Verlag (volume 40). It can be purchased through the publishers. I have often been able to get books from this series through the publication "Microscopy Today". This book gives an extensive list of references about equipment and methods in fluorescence microscopy for standard, confocal and multi-photon microscopy.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Email: gorry2+-at-pitt.edu Name: Michael Gorry School: University of Pittsburgh
Question: Can you recommend any books or journal articles about the basics of fluorescent microscopy photography? I know nothing about cameras and am certain that I could improve the quality of the pictures that I have been taking if I knew more about the camera and exposure times, aperature sizing, etc. It is a Nikon microphot-FXA.
I could find no introductory information from the Nikon people.
Any help would be appreciated.
Thanks
Mike
Michael Gorry E1105 BST University of Pittsburgh Arthritis Institute Pittsburgh, PA 15261
RECEIVED: from SF_Database by POP_Mailbox_-1298363885 ; 15 DEC 98 21:50:11 UT Received: from SPARC5.MICROSCOPY.COM by mailcenter.btny.purdue.edu with SMTP (QuickMail Pro Server for MacOS 1.0.3); 15 DEC 98 21:50:02 UT Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id PAA01759 for dist-Microscopy; Tue, 15 Dec 1998 15:08:29 -0600 Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id PAA01756 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 15 Dec 1998 15:07:58 -0600 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id PAA01749 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 15 Dec 1998 15:07:45 -0600 X-Sender: zaluzec-at-microscopy.com Message-Id: {v03007803b29c82bcad0e-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Date: Tue, 15 Dec 1998 15:19:19 -0600 To: Microscopy-at-Sparc5.Microscopy.Com From: gorry2+-at-pitt.edu () (by way of Nestor J. Zaluzec) Subject: Help on basics of fluorescent microscopy Errors-to: Microscopy-request-at-sparc5.microscopy.com
First of all, thanks a lot for all the answers to my message regarding problems while coating soft cheese samples. I talked to the interested persons about your suggestions, those who also prepare the sample, I do the coating. Indeed, it seems the problem is the temperature raise while coating (a big evaporator adappted for sputtering). On the other hand the sample needs another treatment for drying. Thanks again for spending part of your time writing back to the net!
NEW QUESTION:
This time my question is related to large differences on measuring the width of fibers seen from the surface of different kinds of papers. We have taken micrographs with the SEM of a grid for calibrating at 300X, the error in the calibration was found about 30% (before any calibration correction). At the same time we took micrographs for the different samples at the same magnification, under identical instrument conditions.
On the other hand, the same samples (before and after Au coating) were observed under an optical microscope which was just calibrated. The width of the same fibers showed to be 4 times longer than with the SEM!!!!! Widths in the range of 10 - 20 microns on the SEM measure between 30-80 microns on the OM.
Did any of you happen to have such an uncertainty on measurements? I would trust the OM, but I have no arguments for saying that the SEM is wrong.
I would very much appreciate any comments
Many thanks in advance again.
Silvia Montoro Centro Regional de Investigaciones y Desarrollo Guemes 3450 3000 Santa Fe Argentina csedax-at-arcride.edu.ar
Dear all, has anybody experience cutting silicified chromatographic plates for TEM ultrastructure studies? We are afraid of destroying our diamond knife, is this a serious fear or does it not matter? The silicified graines are 50-60µm laying on a polyethylen sheet. Thanks, Bw
Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit„tsstrasse 25 Germany 33615 Bielefeld phone: 0521 1065592 fax: 0521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
Dear all, has anybody experience or works with in situ hybridization techniques and DNA/RNA negative staining for TEM examinations? Thanks for help , Bw Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit„tsstrasse 25 Germany 33615 Bielefeld phone: 0521 1065592 fax: 0521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
I wish to let you know that due to reasons of the technical and organisational nature it has been decided to shift the Conference Color 1999 to the year 2000 and hold it under the heading of Color 2000. I hope that this "Royal Year" will prove to be more opportune for us - the organizers, and for you - the participants. It will enable us to better prepare this event, while you will have more time to prepare your interesting papers. Due to the above, all the dates stated previously will be shifted by approximately one year, too. Soon we are going to send more detailed agenda.
Wishing you Happy Christmas and Prosperous New Year 1999
I remain
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager of Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
The decision on which microscope to purchase depends on what type of work and samples your lab will be handling. If you will be doing a lot of high resolution work on fixed samples a FEG is great. We have a field emission Hitachi SEM, an S4000. For low voltage work on biological samples it makes fantastic images, but they need to be fixed, dried and coated. They have exquisite detail, but if your sample cannot withstand the preparation or your client is unwilling to pay for all the preparation, the results will be disappointing. I do not have much experience with ESEMs, but from what I have seen, I am not very impressed. They are the ultimate in ease for looking at unfixed, wet samples, but the images from the ones I have seen have looked pretty blah (a technical term). I think this is not just a function of the image formation system in the scope, but of what you are actually seeing, water. With all of the water present in a liquid form, it flows and covers up the surface detail. I think that the combination of a FEG with cryostage would be excellent compromise. It would allow looking at unfixed samples, with everything in place without the time and expense of the preparation, with all of the advantages of the FEG. It would also keep the water were it was at the time of freezing. With sublimation, excess water can be removed, thus exposing surface structures which may have been covered. If you plan on looking at pollen and spore distributions, a cryostage can make all the difference. Bill
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920
-----Original Message----- } From: Debby Sherman [mailto:sherman-at-btny.purdue.edu] Sent: Tuesday, December 15, 1998 6:41 AM To: message to: MSA list
Biological SEM users: I got very little response to the message I posted a week ago (see below). It appears that at the present time there is not a great deal of biological data from FEG-SEMs. To help us determine the best instrument to consider for biological data accumulation, I would also like to explore the pros and cons of the FEI/Philips'ESEM with the low vacuum instruments available from other manufacturers. I have the brochures, etc from a number of companies so am not looking for more of the same. I also have looked into the basic design differences and detector differences.
What I would appreciate is comments from users based on their experiences. Why did you choose the instrument you did? What type of information are you getting from your instrument of choice that you could not get from a conventional SEM? Do you also have a cryo unit and x-ray detector and, if so, what are the limitations for their use in low vacuum modes? How many hours per week is your instrument used and what percentage is in the low vacuum mode? Do you have other conventional SEMs or is this your only instrument?
Any other information would be appreciated. If the response is adequate, and off-line, I will try to summerize.
Thanks, Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
----------------------------------------------- Earlier post (12/7/98): I would like to hear from microscopists who have used FEG-SEM's and particularily the semi-in-lens models for biological applications. I would also appreciate info on recent papers where this instrumentation was critical to the biological research reported. Also any recommendations as to ideal samples for testing these microscopes for biological application? Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
I'm looking for some parts for the epifluorescence system of an Olympus BH2 with the old-style cylinder for holding the filters. Institutional policy requires that we have no equipment less than a century old so Olympus no longer stocks these items. They include:
the little cylinder that looks like part of a submarine periscope, with the filters mounted in a horizontal configuration. I need a spare because my colleagues need the current filters in it (for blue and green excitation), and I would like to do some DAPI. The only number on the cylinder was H92103.
the little "bubble-wand" looking device which is mounted with the filters in a vertical position that slides between two positions for blue and green. Again, I'd like a spare to place my DAPI related filters in.
If anybody has a set of these that they would like to give away, sell, barter, etc. or they know where I might find these, please contact me.
Happy Holidays!
Laura
************************************************************ Yes, it's true. The inmates ARE running the asylum... ************************************************************ Laura Rhoads Electron Microscopy Facility Director Department of Biology Western Kentucky University 1 Big Red Way Bowling Green, KY 42101-3576
Interesting your findings but not a shock to me! SEM calibrations, particularly at low magnification may be at least 25% out. When did you last have the microscope serviced by an experienced technician?
Remember that light and electron imaging reactions do differ. I am not a= n expert on light but I believe I understand what happens with electrons. = I would consider the LM magnifications to be more accurate but read on for why the SEM has problems.
A conventional SEM image at less than 1,000X contains a great deal of BSE=
which either go straight into the Everhart-Thornley detector or are converted to SE by bouncing off components in the chamber. Remember this means sub surface information.
Electron images will have been produced from a volume of material that differs from that of the LM due to penetration through and into the material. You talk of fibres, one assumes they are hairy and that the L= M sees the hairs whilst the SEM does not? If you use the SEM at } 10kV you will certainly have a considerable amount of penetration through the hair= s and as a result you just will not see them. If you or a helper has the skill try 5 or even 2kV when you will see the hairs.
In the SEM business one becomes familiar with seeing things in light that=
do not show up with electrons and of course the reverse, its just part of=
the fun of this super subject.
Good luck
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
} From: "B.Laube-at-biologie.uni-bielefeld.de"-at-Sparc5.Microscopy.Com Organization: Fak. Biologie, Uni Bielefeld To: Microscopy-at-Sparc5.Microscopy.Com Date sent: Wed, 16 Dec 1998 15:10:43 GMT+0100
Dear all, has anybody experience cutting silicified chromatographic plates for TEM ultrastructure studies? We are afraid of destroying our diamond knife, is this a serious fear or does it not matter? The silicified graines are 50-60=B5m laying on a polyethylen sheet. Thanks, Bw
Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=84tsstrasse 25 Germany 33615 Bielefeld phone: 0521 1065592 fax: 0521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
sectioning of any matter is gonna harm a diamond knife with time... the major problem in this case seems to be that some grains will be pulled out of the matrix or away from the substrate, which will a: locally strain the blade normal to the sectioning direction (weak point of every diamond knife) and b: lead to mostly useless sections. I would try to somehow fix the grains (e.g. with a hard embedding material on top) and preferably use an outer edge of a 45 degrees knife at=
fast speed. Alexander Du Chesne, M.S., Ph.D. Max-Planck-Institut f=FCr Polymerforschung PF 3148, 55021 Mainz Tel. 0049 6131 379 195 Fax 0049 6131 379 100 E-mail: duchesne-at-mpip-mainz.mpg.de
1) Why must you make a choice between these different instruments? The FEGESEM allows imaging in either ESEM mode or hivac - user choice. ESEM has come a long way since its beginnings and is worthy of full investigation. There is no need to have surface water present when imaging in ESEM mode. Control of the specimen chamber environment allows an experienced user to remove surface water without dehydrating the sample.
2)The other factor which has not been mentioned when trying to relate OM to SEM is the effect of vacuum on the sample. There has been some nice work on textile fibres by David Taylor in Leeds using SEM and ESEM on the same fibres showing what must be high vacuum effect.
Chris
Chris Gilpin Experimental Officer Biological Sciences EM Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 0161 275 5170 Fax +44 0161 275 5171 Chris Gilpin http://www.empgu.man.ac.uk
Debby;
The decision on which microscope to purchase depends on what type of work and samples your lab will be handling. If you will be doing a lot of high resolution work on fixed samples a FEG is great. We have a field emission Hitachi SEM, an S4000. For low voltage work on biological samples it makes fantastic images, but they need to be fixed, dried and coated. They have exquisite detail, but if your sample cannot withstand the preparation or your client is unwilling to pay for all the preparation, the results will be disappointing. I do not have much experience with ESEMs, but from what I have seen, I am not very impressed. They are the ultimate in ease for looking at unfixed, wet samples, but the images from the ones I have seen have looked pretty blah (a technical term). I think this is not just a function of the image formation system in the scope, but of what you are actually seeing, water. With all of the water present in a liquid form, it flows and covers up the surface detail. I think that the combination of a FEG with cryostage would be excellent compromise. It would allow looking at unfixed samples, with everything in place without the time and expense of the preparation, with all of the advantages of the FEG. It would also keep the water were it was at the time of freezing. With sublimation, excess water can be removed, thus exposing surface structures which may have been covered. If you plan on looking at pollen and spore distributions, a cryostage can make all the difference. Bill
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920
-----Original Message----- } From: Debby Sherman [mailto:sherman-at-btny.purdue.edu] Sent: Tuesday, December 15, 1998 6:41 AM To: message to: MSA list
Biological SEM users: I got very little response to the message I posted a week ago (see below). It appears that at the present time there is not a great deal of biological data from FEG-SEMs. To help us determine the best instrument to consider for biological data accumulation, I would also like to explore the pros and cons of the FEI/Philips'ESEM with the low vacuum instruments available from other manufacturers. I have the brochures, etc from a number of companies so am not looking for more of the same. I also have looked into the basic design differences and detector differences.
What I would appreciate is comments from users based on their experiences. Why did you choose the instrument you did? What type of information are you getting from your instrument of choice that you could not get from a conventional SEM? Do you also have a cryo unit and x-ray detector and, if so, what are the limitations for their use in low vacuum modes? How many hours per week is your instrument used and what percentage is in the low vacuum mode? Do you have other conventional SEMs or is this your only instrument?
Any other information would be appreciated. If the response is adequate, and off-line, I will try to summerize.
Thanks, Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
----------------------------------------------- Earlier post (12/7/98): I would like to hear from microscopists who have used FEG-SEM's and particularily the semi-in-lens models for biological applications. I would also appreciate info on recent papers where this instrumentation was critical to the biological research reported. Also any recommendations as to ideal samples for testing these microscopes for biological application? Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Thanks all, for the reply. Looks like I may have to take matters into my own hands unless "the company" soon up-grades the ccMail v 0.001 :) we are using.
id {Y5R69H3S} ; Thu, 17 Dec 1998 09:15:54 -0600 Message-ID: {7EDF3EF1CA87D211AC160000C0A90FE5115B61-at-engem0.eng.uab.edu} To: Microscopy-at-Sparc5.Microscopy.Com samples!)
Also - when you calibrate and make measurements on the SEM (and TEM) you need to make sure that you are at the eucentric position. Measurement values will change substantially at other heights.
Robin
-----Original Message----- } From: Steve Chapman [mailto:PROTRAIN-at-CompuServe.COM] Sent: Thursday, December 17, 1998 2:14 AM To: INTERNET:"csedax-at-alpha.arcride.edu.ar"-at-sparc5.microscopy.com; American Soc List
Hi,
Interesting your findings but not a shock to me! SEM calibrations, particularly at low magnification may be at least 25% out. When did you last have the microscope serviced by an experienced technician?
Remember that light and electron imaging reactions do differ. I am not an expert on light but I believe I understand what happens with electrons. I would consider the LM magnifications to be more accurate but read on for why the SEM has problems.
A conventional SEM image at less than 1,000X contains a great deal of BSE which either go straight into the Everhart-Thornley detector or are converted to SE by bouncing off components in the chamber. Remember this means sub surface information.
Electron images will have been produced from a volume of material that differs from that of the LM due to penetration through and into the material. You talk of fibres, one assumes they are hairy and that the LM sees the hairs whilst the SEM does not? If you use the SEM at } 10kV you will certainly have a considerable amount of penetration through the hairs and as a result you just will not see them. If you or a helper has the skill try 5 or even 2kV when you will see the hairs.
In the SEM business one becomes familiar with seeing things in light that do not show up with electrons and of course the reverse, its just part of the fun of this super subject.
Good luck
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Dear Woody, We must have a better environment than you; our field was green :-). We too have received several textually-challenged messages. Yours, Bill Tivol
Recently there was a posting inquiring about a source/vendor for a non-magnetic operators chair for EM's. We would be interested in locating a vendor for this type of product.
Bob Roberts Arizona State University Center for Solid State Science Tempe, Arizona 85287
This is a multi-part message in MIME format. --------------9336CF1E225BC7D15F9DF850 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Bob, One positive response to the posting is from Lund.
Mike, We are using the IKEA chair Procent. It works well with the GIF 100. Jan-Olov Bovin
-Mike O'Keefe
Bob Roberts wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Recently there was a posting inquiring about a source/vendor for a } non-magnetic operators chair for EM's. We would be interested in locating a } vendor for this type of product. } } Bob Roberts } Arizona State University } Center for Solid State Science } Tempe, Arizona 85287
--------------9336CF1E225BC7D15F9DF850 Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for O'Keefe, Michael Content-Disposition: attachment; filename="vcard.vcf"
begin: vcard fn: Michael O'Keefe n: O'Keefe;Michael org: {center} {img src=http://mercproj.lbl.gov/logo.gif} {br} {b} {big} National Center for Electron Microscopy {/big} {/b} {/center} email;internet: maokeefe-at-lbl.gov title: Deputy Head x-mozilla-cpt: ;0 x-mozilla-html: TRUE version: 2.1 end: vcard
} Recently there was a posting inquiring about a source/vendor for a } non-magnetic operators chair for EM's. We would be interested in locating a } vendor for this type of product.
This is an issue that many labs are facing, including my own. If there is a commercially available solution (especially in Australia) please let us all know via this listserver.
If not then we are considering doing as an Australian colleague has, i.e. buy a suitably ergonomic chair(s) and have our workshop replace magnetic components with brass. Not a cheap solution but justifiable considering our investment in a GIF.
Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} ---------- } From: Rozeveld, Steve (SJ) } Sent: Tuesday, December 15, 1998 1:00 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: PtSi2 TEM standard } } } } Could anyone suggest a source for PtSi2 ? (powder { 200 micron is OK } ). I only need a few grams. } I thin film (i.e. similar to the NiOx TEM standard) would be ideal. } Yes - I did check with Goodfellows and Aesar. They will } provide the material as a special order but the minimum order was } higher than I needed. } thanks, Steve Rozeveld } } Steve Rozeveld } The Dow Chemical Company } Analytical Sciences Laboratory } 1897 Bldg., Door E43 } Midland, MI 48667 } 517-636-5167 office } 517-638-6443 fax } }
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Mark, Jan-Olov Bovin from Lund writes:
We are using the IKEA chair Procent. It works well with the GIF 100.
I checked on the www and found that this chair seems to use no metal in its construction. I find it is described as: PROCENT swivel chair. Adjustable backrest angle and seat depth. Compression moulded base for improved comfort. Star base and frame of reinforced fiberglass polypropylene foam. Lanna 100% wool in black, blue, dark blue, dark green, dark red, grey or yellow. at: http://www.ikea-usa.com/content/products/prods/PROCENT_CHAIR.asp
On the other hand, I have not tested it, nor even seen one. -Mike
Mark Blackford wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Bob Roberts wrote recently: } } } Recently there was a posting inquiring about a source/vendor for a } } non-magnetic operators chair for EM's. We would be interested in locating a } } vendor for this type of product. } } This is an issue that many labs are facing, including my own. If there is } a commercially available solution (especially in Australia) please let us } all know via this listserver. } } If not then we are considering doing as an Australian colleague has, i.e. } buy a suitably ergonomic chair(s) and have our workshop replace magnetic } components with brass. Not a cheap solution but justifiable considering } our investment in a GIF. } } Cheers, } } Mark Blackford } TEM Group } Materials Division, ANSTO } PMB 1, } Menai, N.S.W. } Australia } 2234 } } Phone 61 2 9717 3027 } Fax 61 2 9543 7179 } } Disclaimer: } The views expressed in this E-mail message do not necessarily represent the } official views of ANSTO from which this message was conveyed.
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begin: vcard fn: Michael O'Keefe n: O'Keefe;Michael org: {center} {img src=http://mercproj.lbl.gov/logo.gif} {br} {b} {big} National Center for Electron Microscopy {/big} {/b} {/center} email;internet: maokeefe-at-lbl.gov title: Deputy Head x-mozilla-cpt: ;0 x-mozilla-html: TRUE version: 2.1 end: vcard
We currently have a Hitachi S570 with a LaB6 filament, however, my management would like to see us get into semiconductor work. For this we would need an FESEM. The questions was asked as to whether or not a standard SEM could be retrofit as with the FE source. My best guess would be "no", but perhaps someone out there has done this - or would know for sure it can't be done. Any information would be welcome (and yes, it would be nice to by a whole new instrument, but.....)
Janet Rice MCC Senior Member Technical Staff rice-at-mcc.com 512-338-3266
We have found two books very helpful, one by Kodak and one by Polaroid.
The one by Kodak is called "Photography Through the Microscope" and is by John Delly. It should be available through most camera stores. We have a few extras here, available for sale, if you can't find it locally.
The other is "Instant Photomicrography Through the Microscope" and should be available through the Polaroid Tech Support hotline (I'm sorry - no contact info - call 1-800-555-1212 for info; they are in Cambridge, MA).
} ... } } } } We currently have a Hitachi S570 with a LaB6 filament, however, my } management would like to see us get into semiconductor work. } For this we would need an FESEM. } The questions was asked as to whether or not a } standard SEM could be retrofit as with the FE source. ...
The answer wouldn't be absolutely not ... after all, FE guns are adapted to similar electron columns. I can think of 3 issues which make the conversion quite expensive (... maybe other can include others ...)
(1) although your LaB6 gun is ion pumped, FE will demand a better ion pump and possibly more efficent 1st stage pumping.
(2) FE guns are said, maybe debatably, to be inherently unstable .. regardless, FE instability is generally compensated for brightness/contrast by an additional beam regulation component.
(3) The resulting increased resolution (... presumably this is why you want FE ...) demands mu-metal electro-magnetic shielding.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I'd like to connect with someone who has tried, successfully (or not), connecting 4pi's SEII card to a Link eXL XP2 pulse processor (PC1934). Please contact me off line at the address below. TIA.
Owen
============================= Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
Cryo TEM Service Work. I need a lab that is doing outside contract cryo TEM or cryo TEM replica work. I have some samples th= at I need prepared and photographed.
Contact: Joanne M. Crudele Unilever HPC USA Joanne.Crudele-at-Unilever.com or 847-734-3712 =
A simple Pappenheim-stain (May-Grunwald/Giemsa) is probably the simplest way to make a "quick blood stain to differentiate...".
This stain is easy to perform, fast (about 10 - 15 min) and gives very good differentiation of all blood-components!
Protocols can be found in any textbook on basic histological technique. If you can't find them, let me know...
Yvan Lindekens. ---------- } Van: Jim J Darley {jim-at-proscitech.com.au} } Aan: Microscopy-at-sparc5.microscopy.com {Microscopy-at-Sparc5.Microscopy.Com} } Onderwerp: blood stain; DNA vs cytoplasm } Datum: dinsdag 15 december 1998 11:17 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------- } I received an inquiry asking what would be a most suitable } stain } for student's to perform a quick blood stain to } differentiate } lymphocytes, macrophages, neutrophils etc (basically a DNA } vs cytoplasm } stain).
In addition to RCHIOVETTI-at-aol.com message: another interesting brochure on fluorescent microscopy was made by Reichert in Vienna (now also Leica):
"Fluorescence microscopy with fluorochromes - recipes and tables"
A good chapter on fluorescent microscopy photography can be found in:
C. Van Duyn Jr: "Mikrofotografie", Focus Elsevier, The Netherlands (no ISBN). This is in Dutch.
Some intresting notes on the subject can also be found in:
Goke, G: "Modene Methoden der Licht-mikroskopie", Kosmos Wissensschaft, Franckh, Stuttgart 1988, ISBN: 3-440-05765-8. This is in German.
Hope this helps...
Yvan Lindekens.
---------- } Van: RCHIOVETTI-at-aol.com-at-Sparc5.Microscopy.Com } Aan: gorry2+-at-pitt.edu; Microscopy-at-Sparc5.Microscopy.Com } Onderwerp: Re: Help on basics of fluorescent microscopy } Datum: woensdag 16 december 1998 5:25 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } In a message dated 98-12-15 16:38:37 EST, gorry2+-at-pitt.edu writes: } } { { Question: Can you recommend any books or journal articles about the basics } of fluorescent microscopy photography? } } } } Mike, } } There is a small book that was produced by Wild Leitz in Germany, entitled...
Hi, all We're looking for an LWD 40x phase objective and some filter cubes to go with a used Nikon Diaphot with epi-fluorescence. If you have one or more of these items, please contact me at the e-mail address below; please do not reply to the list.
The subject line says nearly all of it. We're looking for a 40x long-working distance phase objective with correction collar for a Nikon diaphot: Any one of the following three objectives will do: } CF Achromat LWD DL 40x C (78816) } CF N Plan Achromat ELWD DM 40xC (85053) } CF Fluor LWD DM 40x C (85008)
We are also looking for blue, green and UV cubes for an epifluorescence illuminator; standard Nikon will do, as these are going in an Opti-Quip slider
TIA Julian Smith III Biology Winthrop University Rock Hill SC 803-323-2246 (fax) 803-323-2111 x6427 (vox) smithj-at-winthrop.edu
Julian P.S. Smith III Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x6427 (vox) 803-323-2246 (fax)
-----Original Message----- } From: Doug Danielson [mailto:wddanie-at-ibm.net] Sent: Saturday, December 19, 1998 1:48 PM To: Microscopy-at-MSA.Microscopy.Com
Hi Mark, I must be getting real old as I remember my gransfather used to make chairs out of plant material. Large plants I believe. They wood cut the larger stems into structural members and glue the parts together into many different shapes. I sure these were not as functional or beautiful as the bent steel and plastic used today. Maybe you could find one of these antiques for your purpose. Russ
-----Original Message----- } From: Mark Blackford [mailto:mgb-at-ansto.gov.au] Sent: Friday, December 18, 1998 10:15 AM To: Microscopy-at-sparc5.microscopy.com
Bob Roberts wrote recently:
} Recently there was a posting inquiring about a source/vendor for a } non-magnetic operators chair for EM's. We would be interested in locating a } vendor for this type of product.
This is an issue that many labs are facing, including my own. If there is a commercially available solution (especially in Australia) please let us all know via this listserver.
If not then we are considering doing as an Australian colleague has, i.e. buy a suitably ergonomic chair(s) and have our workshop replace magnetic components with brass. Not a cheap solution but justifiable considering our investment in a GIF.
Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} From January 1999 there will be a 3 year post-doc position available in the electron microscopy group (Department of Physical Chemistry) at the University of Mainz, Germany. At present the group has 2 electron microscopes and a well-equipped specimen preparation laboratory. In the summer of 1999 a new 300 kV Philips Tecnai 30 Electron Microscope with FEG and GIF will be installed. Applications from physicists and chemists with Ph.D experience in electron microscopy and a sound background in electron energy loss spectroscopy are welcome.
Please send applications to
Dr. I.G. Voigt-Martin Inst. f. Physikalische Chemie Jacob Welder Weg 11 55099 Mainz
Hello, I have one Sorvall MT 5000 ultramicrotome that is in operating condition, and a Durst Laborator (185) enlarger with all the lens' still operational, for sale. If ineterested, E-mail or call with an offer.
The Matic-Mot2 camera controller for my Leco 300 has died - it gives a "Camera?" message and doesn't function. Anybody out there have this happen? Any known source to have it fixed? The Leco folks say that parts are very rare and the cost to fix would be $2-4K (USD).
If anyone has a spare Matic-Mot2 camera controller or knows of a fix, let me know.
I decided to hang a Polaroid DMC on the microscope, but would still like to have the capability to take 4X5 Polaroids.
Happy Holidays,
Barry Ritchey mbritch-at-sandia.gov Sandia Nat'l Labs - NM
While we don't have a used polishing wheel, we do offer an inexpensive 8"=
wet polishing wheel (Model 900) which is less than $1,000. For informati= on you can see our web site at www.southbaytech.com or give me a call at 800-728-2233.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by INTERNET:"DChernoff-at-aol.com"-at-sparc5.microscopy.co= m } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
I'm looking for a used, inexpensive wet polishing wheel. Contact Don Chernoff, (703) 849-1492, dchernoff-at-aol.com
A while back someone asked about automatic tissue processers. I don't recall seeing much responce. I have been asked to investigate pros and cons for our pathology dept. TEM lab. I have literature on RMC and Leica. We looked at one ten years ago and our only concern at that time was possible drying artifact from tissue being lifted out of the propylene oxide, and that there were not many people using them. Thanks for any help.
Rick Vaughn Univ. Nebr. Med. Ctr. RLVAUGHN-at-MAIL.UNMC.EDU
Wam wszystkim z ktorymi wspolpracuje z okazji Swiat Bozego Narodzenia i Nowego Roku 1999 skladam zycenia Duzo Zdrowia i Wszystkiego Najlepszego dla Was i Waszych Najblizszych.
Niech kazdemu spelni sie przynajmniej jedno z tych cichych marzen ktore, gdy wieczorem zasiada sie w fotelu to wraca - a gdy tak ....
Wszystkiego Najlepszego
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager of Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
I am going to work with TEM JEM-2010X, but to start is hard for me because of manuals, that are written in Japanese. I need to make a chemical composition analysis of about 3x3 micron area on semiconductor surface. And the only installation for such an analysis with appropriate spatial resolution is JEM-2010X. To simplify the procedure I'd like to avoid the common routine of sample preparations for TEM. But for that I need to know if the mentioned installation has such an option to observe the surface in non-transmission way.
Any suggestions on where to get the manual in English (printed, or in any file format) would be highly appreciated.
Dmitri.
__________________________________________ Dmitri V. Sokolov Doctor Course student Research Center for Interface Quantum Electronics, Hokkaido University, North 13, West 8, Kitaku, Sapporo 060, Hokkaido, Japan
Phone 81-11-706-7174 Fax 81-11-716-6004 Home phone 81-11-261-2185
If you have any wishes I wishe you to have them made true !!!
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager of Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
On Wed, 23 Dec 1998 VANDERHAEGHEN_JACQUES/CENTER_RD-at-bekaert.com wrote:
} Please if you send a e-mail do it in Englisch. } But, I want to send you the best wisches for Christmas and a very happy } new year. } } Jacques Vander Haeghen } Belgium } ---------- } } Kochani, } } } Wam wszystkim z ktorymi wspolpracuje z okazji Swiat Bozego Narodzenia i } Nowego Roku 1999 skladam zycenia Duzo Zdrowia i Wszystkiego } Najlepszego dla Was i Waszych Najblizszych. } } Niech kazdemu spelni sie przynajmniej jedno z tych cichych marzen ktore, } gdy wieczorem zasiada sie w fotelu to wraca - a gdy tak .... } } Wszystkiego Najlepszego } } } Krzysztof Jan Huebner } } {hubner-at-IOd.krakow.pl} :-) } } FOUNDRY RESEARCH INSTITUTE } Research Materials Department } Manager of Structural and Physical Research Laboratory } str. Zakopianska 73 Call (*48 12) 2665022 ext.356 } 30-418 KRAKOW - POLAND Fax (+48 12) 2660870 } } }
I have been using a RMC processor for 3 - 4 years now. It is used on a = daily basis, yes a few problems and yes a few lost specimens. If = possible do not process all of biospy received. I like to RMC because of = the individual caps to reagents. Have not had any problems do to = evaporation of solvents. =20
Best of Luck,
=20
Ed Calomeni Dept. Pathology Medical College of Ohio 3000 Arlington Avenue Toledo, Ohio 43614
Just to start the New Year off on an up beat: I would like to announce:
The Fourth International Short Course on 3D Microscopy of Living Cells June 16 - 27, 1999
and
The Post-course Workshop on 3D Image Processing June 29- July 1, 1999
in association with the BioSciences Microscopy Facility and Department of Computer Science
University of British Columbia, Vancouver, BC, Canada
Organized by Prof. James Pawley: University of Wisconsin-Madison
We expect to have at least 11, 3D microscope workstations, including several using multi-photon excitation, for student use and there will be an international faculty of 14. The schedule is basically lecture/demonstration in the mornings, labs in the afternoons and personal, live cell projects at night. Many contributors to this list are "alumnae."
APPLICATIONS Applicants will complete a questionnaire to assess knowledge level, field of interest and proposed personal, live-cell, project. Enrollment will be limited to about 24 participants. Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with basic texts to read before the course begins. Application forms requesting information on field of interest and level of experience may be down-loaded from the WWW site at
http://www.cs.ubc.ca/spider/ladic/course/forms/register.txt or obtained from:
Prof. James Pawley, Rm. 1235, 1117 W. Johnson Ave., Madison, WI, USA 53706. Phone: 1-608-263-3147, Fax 1-608-265-5315, Email: jbpawley-at-facstaff.wisc.edu
Application deadlines:
Application forms must be received for screening by March 1, 1999. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 1999. In general, refunds of the deposit will not be possible. The remainder is due before Registration.
DATES: Applications must be received by Mar. 1/99 50% deposit due Apr. 15/99 Registration 4:00 - 7:00 pm Wednesday, June 16, 1999 Last class will end with lunch Sunday, June 27, 1999
3D Image Processing Workshop June 29- July 1, 1999
I realize this is not the best time to post a question to the list, with so many people unsubscribed for the holidays, but here goes. The question has to do with the generation of Cathodoluminescence (CL) in specimens as a function of incident electron energy. Leverenz (1968) has a number, apparently a calculated range, of 0.2 microns for a 6 keV electron in Zn2SiO4 and says that the electron range is proportional to the square of the electron energy in this range. Is there any good more recent data on the penetration depth of electrons in minerals, e.g., silicates, carbonates, as a function of the electron enery,in the range of 5kV to 100 kV?
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Happy Holidays, Does anyone know of a company or small contractor who will service Reichert microtomes (other than the more expensive bigger companies like Leica). I recall a company called Tek-net in N.J. but am unsure. Salutaions,
I know it's Christmas day, but I have opened all my prezzies and found your message about penetration depth. You can calculate the average depth of electron peneration using the Bethe range equation. See equation 3.13 Page 88 of SEM-XRMA by Goldstein,Newbury,Echlin,Joy,Romig,Lyman,Fiori & Lifshin. lenum Press NY 1992 which gives all the good stuff. Also use Electron Flight simulator a piece of software which does all the sums for you.
Happy Christmas.
I'm back to the claret and Stilton
Patrick Echlin Multi-Imaging Centre Cambridge University United Kingdom
On Thu, 24 Dec 1998, MICHAEL DELANNOY wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Happy Holidays, } Does anyone know of a company or small contractor who will } service Reichert microtomes (other than the more expensive bigger } companies like Leica). I recall a company called Tek-net in N.J. } but am unsure. Salutaions, } } Mike D } } } }
Happy Christmas everybody Does anybody have an unwanted liquid nitrogen-cooled cold stage suitable = for a Philips 430 TEM, with retractable protective blades? We have = temperature control and transfer systems for Oxford and Gatan systems on = other TEMS, so may only need the specimen holder itself. But we are = looking for the type of holder that can be used to transfer a cold = specimen into the microscope, rather than just cool it down within the = column.=20 Yours in hope Sally
Dr Sally Stowe Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475 ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525
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You may also try MOC (914-268-6450), we have been using them for a couple of years and they are OK.
-- Michal Jarnik, Ph.D. Fox Chase Cancer Center Electron Microscopy Facility 7701 Burholme Ave. Philadelphia PA 19111 Tel. 215-728-5675 Fax 215-728-2412
--------------7DCF927DFE885DC8910A00A0 Content-Type: text/x-vcard; charset=us-ascii; name="M_Jarnik.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Michal Jarnik Content-Disposition: attachment; filename="M_Jarnik.vcf"
begin:vcard n:Jarnik;Michal x-mozilla-html:FALSE adr:;;;;;; version:2.1 email;internet:M_Jarnik-at-fccc.edu x-mozilla-cpt:;3 fn:Jarnik, Michal end:vcard
Hello.=20 My name is choong keun Yoo and graduate student at hanyang university in korea. I have a question for sample preparation in cross-sectional TEM. i will explain my problems following paragraph.
At first, using Si dummy wafers outside of two ribbons along with epoxy, Si dummy wafers were broken in cutting with diamond saw. So next step is to use moly and copper tubing kit. (ref. attached file) but next problem is that specimen's thickness is too slim to fix to moly and copper kit.=20 And the material putting in the space is so difficult to make and they( side material and specimen) are different in dimpling and ion milling rate. So prepered hole is generated in dimpling. (ref. attached file) So far, i said my problems in experiment
How can i promote my experiment? What method is good for that case? Wanna your help...
Can one of you out there on the listserv suggest a training program for electron microscopy of semiconductors? I'm looking for training in both sample prep techniques and in SEM technique as well. If we aren't likely to upgrade equipment at the moment, upgrading skills would not be a bad idea.
Janet Rice MCC Senior Member Technical Staff rice-at-mcc.com 512-338-3266
We run courses "in house", in your own laboratory on your own equipment, = on all aspects of electron microscopy. Please take a look at our web site f= or more information, we are able to tune a course to any requirement. =
Advantages of using your own equipment are that we are able to see problems that may be due to that particular equipment. AIso imaging on different systems, different specimen detector geometry in the SEM, may give considerably different results,
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
hi: I'm looking for tissue processing technics using microwave in SEM and TEM. any help is welcome
thanks in advance =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D Fernando D. Balducci Laboratorio de Microscopia Electr=F3nica Facultad de Ingenier=EDa - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
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I am looking for information on techniques using optical microscopy to quantify flaws within hardened concrete and grout samples. I am currently using Image Pro 3.0 with the Materials Pro module to quantify images off of a video microscope and am looking for more in-depth information. Thanks.
I would like to sell magnetically levitated turbomolecular pump based - TPU 180, TCM 180, MD4T Balzers - oil-free pumping system ($20420.- original cost a year ago). Alternatively, I would be interested in trading this system in for cryo-pumps. I will be happy to provide more detailed information to or to discuss offers with those who are interested. Marek Malecki.
All interested are invited to apply! This announcement can also be found at: http://www-biology.ucsd.edu/positions/molGenBioChemCell.html
Manager and Electron Microscopist Biology Department Electron Microscopy Facility University of California, San Diego, La Jolla, CA Job title: Associate Specialist (salary range $42,660-$45,624)
A position is available January 1, 1999, for an individual with extensive expertise in the preparation and analysis of biological materials for electron microscopy. A Ph.D. or M.S. degree in the life sciences with substantial graduate research experience in electron microscopy and a high level of skill in preparation and analysis of samples by transmission electron microscopy are required. Experience with the following techniques is preferred: negative staining and immunogold labelling for TEM, cryofixation and cryosectioning, scanning electron microscopy, confocal microscopy, developing and printing of photographic negatives, and preparation of digital images.
The individual hired will have primary responsibility for all aspects of the Biology Department Electron Microscopy Facility's operation. Essential functions include performing electron microscopy on a fee-for-service basis for a wide range of scientific projects, as well as training and assisting users of the Facility in sample preparation and analysis. The Manager will oversee maintenance of equipment and the Facility in general, as well as record keeping, ordering, and upkeep of supplies.
The facility serves a community of active UCSD and biotech scientists. The Manager of the EM facility will interact with top EM facilities in the La Jolla area. Opportunities for grant writing and involvement in growth of the facility are possible.
Closing date for applications is February 15. Inquiries about the position may be sent to:lsmith-at-biomail.ucsd.edu. Send applications (CV with names, addresses, phone and Fax numbers and email addresses of 3 references) to:
Carrie Garnett Supervisor, Academic Affairs Department of Biology - 0346 UCSD 9500 Gilman Drive La Jolla, CA 92093-0346
UCSD is an equal opportunity/affirmative action employer with a strong institutional commitment to the achievement of diversity among its faculty and staff.
Laurie G. Smith Assistant Professor Biology Dept. 0116 U.C.San Diego 9500 Gilman Drive La Jolla, CA 92093-0116 email: lsmith-at-biomail.ucsd.edu telephone: 619-822-2531 (office) 619-822-2558 (lab) fax: 619-534-7108
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
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Microscopy/Microscopy Education just added Lisa Montanaro to our staff. She has nearly a decade of experience in this area, having worked for IBM and for Dominion. (She was given the "Best of the Best" award at IBM). Since we specialize in integrating your level of experience, with your equipment and your application, we can tailor a course specifically to fit your needs. Our offices will be open again next Monday. Please call and we can discuss details.
The TEM Service Group in the Chemistry and Materials Science Directorate at Lawrence Livermore National Laboratory (LLNL)is considering posting a term position for an Electron Microscopists. The position will be open to all candidates with a technical or professional degree and 1-2 yrs. of related work experience.
The primary job function will be to perform a variety of (S)TEM techniques (as needed) to characterize the microstructure of a wide variety of materials that are utilized and/or are being developed for internal and external programs that LLNL supports. Examples include: Multilayered materials, areo-gels, metallurgical materials, ceramics, etc. Additional duties will include the use and developement of sample preparation techniques for TEM. Basic level experience or working knowledge of conventional, analytical and HREM TEM techniques and associated analysis techniques is required. Knowledge of materials science, metallurgy, computers, software, etc. is desired. USA citizenship is preferred.
The position will be a 1yr term, renewable up to 6 years. The salary range will be =89 $3.5 - 4.5K / month.
Send inquires and/or resumes to:
Mr. Mark A. Wall Sr. Scientific Assoc. L-350 7000 East Ave. C&MS Dir. Lawrence Livermore National Laboratory Livermore, CA USA 94551
or
Email WALL1-at-LLNL.GOV
Mr. Mark A. Wall Sr. Scientific Assoc. L-350 Chemistry & Materials Science Directorate Lawrence Livermore National Laboratory Livermore, CA USA 94550
ph: 925 423-7162 fax: 925 422-6892
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