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Lots of pixels mean a larger chip. At the side mount position, the diameter of the electron beam is perhaps 1-2 cm, so with a large chip, you would only be illuminating a part of the chip. So, no point in having a chip with lots of pixels at the side mount position.
There are at least two suppliers of 4k by 4k TEM CCD cameras - that is, 16 megapixels, so you might want to do a bit more research on your options.
I would also be careful in use of the word resolution - lots of pixels does not necessarily mean that you get good resolution in the final image. The key factor is the point spread function. To transfer the electron image to the CCD, there is a scintillator and then a coupling mechanism - either lenses/prism or fibre optic. What needs to be considered is how wide an illuminated area is generated at the CCD by a single electron hitting the scintillator. If you have small pixel elements in the CCD, you end up with a single electron illuminating 4 or 9 pixels - in such a case, there would be no advantage to having lots of pixels. Very large pixel elements will result in electrons at different positions in the image illuminating the same CCD pixel element. So, good TEM CCD camera design requires a careful matching of the CCD pixel element size to the point spread function. Given that this match has been achieved, large pixel elements are good - they store more charge, thus giving you greater dynamic range.
I guess what that boils down to is, don't assume more pixels means a better image. If the camera is well designed, it should but it is not an assumption I would make. You also need to think about read-out time - more pixels take longer to read-out to the computer.
My advice would always be to get images from your specimens and compare the results from different cameras.
Here is my protocol for HMDS drying of bacteria. Have fun!
Dave
Preparation of bacteria for SEM using HMDS
Safety Work in fume hood and use gloves Retain ethanol and HMDS (hexamethyldisilazane) for disposal in non-chlorinated waste bottle
Start with a lot of bacteria eg a 3 x1mm cubed pellet
Fixation
Fix in 4% glutaraldehyde in buffer (usually 0.1M )* Leave for 1 hr at room temperature or 24hrs in the fridge Rinse in buffer x3 (total storage time should exceed fixation time by a factor of 3).
SEM Dehydration of bacteria in an Eppendorf
5m in 20% ethanol 5m in 30% ethanol 5m in 50% ethanol 5m in 70% ethanol 5m in 90% ethanol 5m in 100% ethanol 5m in 100% ethanol
5m in 100% ethanol / HMDS (2:1) 5m in 100% ethanol / HMDS (2:2) 5m in 100% ethanol / HMDS (1:2) 5m in 100% HMDS 5m in 100% HMDS
Centrifuge and resuspend in a 2 drops" of HMDS. Put on drop on a clean (use acetone) small round coverslip on a stub. Take rest of sample dilute by a drop and mount one drop. Do this about 4 times to get a range of dilutions.
Sputter with gold and view in SEM.
*For 4% glutaraldehyde in 0.1M buffer Take, say, 50mls of 0.2M buffer, 34ml d water and 16ml of 25% glutaraldehyde.
On Wed, 26 Feb 2003 10:49:26 -0600 (CST) "ggm-at-servidor.unam.mx"-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 } } } I NEED TO DRY BACTERY BY SUBLIMATION USING HEXAMETHYLDISILAZANE } (HMDS), SOMEONE, COULD HELP ME, BECAUSE ITS THE FIRST TIME I USE IT. SOMEONE } COULD TELL ME HOW TO USE HMDS. } } THANKS. } } M.C. GUILLERMINA GLEZ. MANCERA } FAC.QUIMICA-UNAM } MEXICO, D.F. } } ------------------------------------------------- } Obtén tu correo en www.correo.unam.mx } UNAMonos Comunicándonos } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
} My thanks to everyone who has sent me advice and information on digital } image systems for TEM. I am still having trouble understanding why their } is a difference in resolution between side mounted cameras and bottom } mounted cameras. I see that bottom mounted cameras have a resolution of up } to 6 megapixels while the highest I have seen so far for side mounted } cameras is 2 megapixels. If anyone out there would be willing to try and } explain this to me I would appreciate this and then I could in turn pass } that on to my faculty bosses who would also like to know about the } difference. } } Tom Bargar } Electron Microscopy Core Research Facility } 986395 Nebraska Medical Center } Omaha, NE 68198-6395 } } phone 402-559-7347 } } tbargar-at-unmc.edu
Lots of pixels mean a larger chip. At the side mount position, the diameter of the electron beam is perhaps 1-2 cm, so with a large chip, you would only be illuminating a part of the chip. So, no point in having a chip with lots of pixels at the side mount position.
There are at least two suppliers of 4k by 4k TEM CCD cameras - that is, 16 megapixels, so you might want to do a bit more research on your options.
I would also be careful in use of the word resolution - lots of pixels does not necessarily mean that you get good resolution in the final image. The key factor is the point spread function. To transfer the electron image to the CCD, there is a scintillator and then a coupling mechanism - either lenses/prism or fibre optic. What needs to be considered is how wide an illuminated area is generated at the CCD by a single electron hitting the scintillator. If you have small pixel elements in the CCD, you end up with a single electron illuminating 4 or 9 pixels - in such a case, there would be no advantage to having lots of pixels. Very large pixel elements will result in electrons at different positions in the image illuminating the same CCD pixel element. So, good TEM CCD camera design requires a careful matching of the CCD pixel element size to the point spread function. Given that this match has been achieved, large pixel elements are good - they store more charge, thus giving you greater dynamic range.
I guess what that boils down to is, don't assume more pixels means a better image. If the camera is well designed, it should but it is not an assumption I would make. You also need to think about read-out time - more pixels take longer to read-out to the computer.
My advice would always be to get images from your specimens and compare the results from different cameras.
The University of Texas Health Science Center and Hamamatsu Photonics KK will co-host A Special Topics Symposium and Workshop on
Fluorescence Lifetime Imaging and Spectral Imaging
Symposium - Academic and Commercial Presentations June 6-7, 2003 Gunter Hotel Student: $ 200.00 (US) Professional/Corporate: $ 250.00 (US) Registration Deadline May 1, 2003
Workshop - Hands-on Experience with Latest Instrumentation June 8-10, 2003 UT Health Science Cnt. Tuition: $700.00 (US) Includes room and board 20 student limit / 4 scholarships available Application Deadline April 7, 2003
For information and forms visit:
http://usa.hamamatsu.com/flim_spectral/default.htm or http://www.uthscsa.edu/csb/imaging-course.html
The -lm switch solved the problem. The math libraries need to be explicitly linked on the gcc command line. It is also not necessary to include tgmath.h. I have successfully compiled atompot, mulslice and image programs and they run successfully on the sample data files provided with the source code. The commands I used were:
My thanks to Paul Voyles and Dimiter Prodanov for their suggestions. The suggestion from Dimiter Prodanov was to upgrade to gcc 3.0 and not to use the -ansi option. I have not used the -ansi option but upgrading to gcc 3.0 is not required for this particular set of programs, IMHO.
With Best Regards, Divakar
---- Dr R Divakar Physical Metallurgy Section MCG-IGCAR, Kalpakkam 603102, India ----
-----Original Message----- } From: Paul Voyles [SMTP:voyles-at-engr.wisc.edu] Sent: Thursday, February 27, 2003 10:02 AM To: Divakar R (Dr) Cc: Microscopy-at-sparc5.Microscopy.Com
Hi everybody,
does anybody know anything about metrizamide embedding of TEM specimens?
Philip
Philip Koeck Södertörns Högskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em
What I remember from an immunogold workshop given by Moise Bendayan, University of Montreal, Canada, is that different antigens react to osmium differently. Some are OK and others are not. If no one had worked on it before you have to find out yourself. You may incubate osmium treated section on a drop of saturarated sodium metaperiodate for 1 hr. before immunostaining. Following reference covers tissue precessing. Bendayan, M. Protein A-gold and Protein G-gold postembedding immunoelectron microscopy. In Colloidal Bold: Principles, Methods and Applications Vol. 1, p33-94. Acacemic Press 1989.
AnnFook yang
} } } Tamara Howard {thoward-at-unm.edu} 02/28/03 01:22PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mike -
I'm sure there are references out there, but all I can offer is personal experience and word-of-mouth. I've used 1 (that I can think of off the bat) antibody that gave "normal" staining on sections of tissue with regular osmication, and some that were OK with reduced osmium (osmium mixed with potassium ferrocyanide or ferricyanide - you'll get reams of info on which to use!), and enough that were useless after any osmium to make you cry. Osmium can also mess up your resin polymerization, so you have to be careful - especially if you want to UV cure - dark tissue is a major pain.
What do you mean by osmium reducing the size of gold label? You said you were doing postembedding immuno....I haven't heard of any gold size problems for postembedding (lots with pre-embedding and osmium) - how would that happen?
Do you have some potentially unrescuable blocks that have already been osmicated? You might try a gentle peroxide treatment on the sections, or some EM version of antigen retrieval....
Good luck!
Tamara
On Fri, 28 Feb 2003, Mike 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 } -----------------------------------------------------------------------. } } } To the listserver, } I am looking for references on the effects of osmium staining } on postembedded immunolabeling, ie reducing the amount and size } of gold label on LR Gold/Whites sections. Also has anyone } used very low osmium (0.05%?) successfully for IEM? } } Thank You } Mike D } } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
Anyone have a manual for a Nikon Labophot Pol? I need to do a little cleaning and probably regreasing for it, but don't have any kind of a manual, basic, service or otherwise. Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
We've lost the operating instructions to our LKB 2178 Knifemaker II. Can someone make us a copy of their's? I'd certainly compensate you for the associated costs. Thanks in advance.
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-487-2002 FAX 906-487-2934 Mailto: opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills.htm
The following party is looking for a outside laboratory to study the structure of DNA in plasmid. This is beyond the scope of our laboratory. I told her that someone in MSA can help her. J. Roy Nelson, Ph.D. Material Testing Laboratory mtl-at-njcc.com
"I am interested in using electron microscopy to look at supercoiled versus open circular DNA in our sample of purified plasmid DNA. I would like to know who might offer this service and their price for the service."
Has anyone done TEM on Armadillidium vulgare (Pillbugs)? We're interested in the structure of the eyes of these pillbugs. We're not very sure about the structures that we are seeing and the preparation of the specimen. If anyone has any literature that they can recommend or experiences that they can share I would greatly appreciate the help.
Khara Scott EM Technician Chicago State University
__________________________________________________ Do you Yahoo!? Yahoo! Tax Center - forms, calculators, tips, more http://taxes.yahoo.com/
Refer to "G. M. Brown and J. H. Butler, "New method for the characterization of domain morphology of polymer blends using ruthenium tetroxide staining and low voltage scanning electron microscopy (LVSEM)", Polymer 38 (15), 3937 (1997)" for a detailed description for preparation of RuO4 solutions.
Regards,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"George.Theodossio u-at-csiro.au" To: Microscopy-at-sparc5.microscopy.com cc: Subject: TEM Polymer Staining Procedure 02/13/03 01:14 AM
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Dear All,
I have a colleague who is looking at silica particles in a polymer matrix. He is planning TEM analysis of cross sections to image the particles, determine their distribution and possibly some EDXS mapping. The samples are cast as thin films and the silica could be as small as 5nm. He has found a reference to the use of Phosphotungstic Acid (hope the spelling is correct) to stain the polymer but not the silica, thus improving contrast in the TEM. This was done by floating off the cryo-microtomed sections on a methanol solution of Phosphotungstic Acid. Alas, this is all the information he has.
We don't have cryo microtomy facilities, so we were going to use our microtome at room temp and also attempt to tripod polish.
Does anyone have a more detailed procedure of how to make up the solution and stain the polymer?? Any assistance would be greatly appreciated.
Regards George
George Theodossiou Physicist / Electron Microscopist CSIRO Manufacturing & Infrastructure Technology Private Bag 33 Clayton South MDC Victoria, 3169 tel: +61 3 9545 2012 fax: +61 3 9544 1128
To the extent permitted by law, CSIRO does not represent, warrant and/or guarantee that the integrity of this communication has been maintained or that the communication is free of errors, virus, interception or interference.
The information contained in this e-mail may be confidential or privileged. Any unauthorised use or disclosure is prohibited. If you have received this e-mail in error, please delete it immediately and notify George Theodossiou on +61 3 9545 2012. Thank you.
I am a book designer with the University of Illinois Press and am searching for a SEM image of a cancer cell(s) to use on the cover of a book we are publishing.
It was suggested that I make my query on this list. The book talks specifically about mantle-cell diffuse lymphoma but the image does not need to be that exact, any malignant lymphoma would do.
We would of course credit any images we use and perhaps be able to pay you or your organization a small fee. Please email me direct if you have any information to pass on.
Thank you,
Paula Newcomb Designer University of Illinois Press 217.244.4706 newcomb1-at-uiuc.edu
Below is the Table of Contents from the upcoming issue of the Journal of Microscopy & Microanalysis. Vol 9, No. 2
Editorial Charles Lyman
Historical Bibliography Key Events in the History of Electron Microscopy F. Haguenau, P.W. Hawkes, J.L. Hutchinson, B. Satiat-Jeunemaître, G.T. Simon, and D.B. Williams
Papers from the Australian Microbeam Analysis Society
Monet's Painting under the Microscope Paula Dredge, Richard Wuhrer, and Matthew R. Phillips
Cathodoluminescence Efficiency Dependence on Excitation Density in n-Type Gallium Nitride Matthew R. Phillips, Hagen Telg, Sergei O. Kucheyev, Olaf Gelhausen, and Milos Toth
Application of Quantitative Electron Microscopy to the Mineral Content of Insect Cuticle Ron Rasch, Bronwen W. Cribb, John Barry, and Christopher M. Palmer
Charge-Related Problems Associated with X-Ray Microanalysis in the Variable Pressure Scanning Electron Microscope at Low Pressures Brendan J. Griffin and Alexandra A. Suvorova
Dear List, I'd like to find a protocol for making colloidal gold in 10 nm and 20 nm sizes. I'm sure someone on the list has either a procedure or a reference to one. TIA for your help. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Dear Colleagues, The 4th ASEAN Microscopy Conference (ASEANMC4) will be held in Hanoi, VietNam, from October 9 till 10, 2003. This Conference will be a gathering of microscopists from the ASEAN and other regions of the world. An important aim of the Conference is to promote mutual understandings and to develop international collaborative research in life and material sciences. Participants will have the opportunity to review recent work, to discuss recent advances and identify new challenges in the field of electron and light microscopy. The Second announcement are currently in the mail to many microscopist who are interested in this conference. When we receive reply from you then we will send "the Second announcement" to you by post as soon as possible. We would like to extend our warmest welcome to you to participate and look forward to seeing you in Hanoi in October 2003.
Chairman, Organizing committee
Prof. Nguyen Van Man
***********************************************
Mailing address:
Assoc. Prof. Nguyen Kim Giao
Electron Microscopy Unit
National Institute of Hygiene and Epidemiology
N0 1, Yersin street- Hai Ba Trung district - Hanoi - Vietnam
Tel: 84.04.9715434 Fax: 84.04.8210853
Email: {mailto:emlad-at-hn.vnn.vn} emlad-at-hn.vnn.vn, or {mailto:emunihe-at-vol.vnn.vn} emunihe-at-vol.vnn.vn
Why make your own gold, so much easier to buy and store premade?
Anyway, way back when there was a review article in the EMSA Bulletin entitled Colloidal Gold as a Marker and Tracer in Light and Electron Microscopy. Published around 1986??? Author was J. Dewey This is a nice review article on most aspects of colloidal gold labeling..
Other references to look up are:
1). Mulpferdt, H. Experientia, 1982 (38) pg 1127-1128 2). Roth, J. 1983b Techniques in Immunocytochemistry, Vol. II (Academic Press) pg 217-284.
best of luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
Note to list: This E-mail included a rather lengthy attachment that cannot be sent via the list. Please E-mail me directly if you want the procedure for making colloidal gold conjugates.
Bill, I have used the enclosed procedure many times and it works great. Although I am lazy these days and usually buy gold conjugates, I still have some prepared by this method that worked great 8-10 years after being made. I made primarily 5 and 10nm gold so cannot vouch for the 20nm results.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 3/3/03 5:52 PM, "Bill Tivol" {tivol-at-caltech.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, } I'd like to find a protocol for making colloidal gold in 10 nm and 20 } nm sizes. I'm sure someone on the list has either a procedure or a } reference to one. TIA for your help. } Yours, } Bill Tivol } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 } tivol-at-caltech.edu } } }
I kindly ask you for your advice concerning inverted microscope. We need to count tiny mites in alcohol samples. Mites were washed from plants, so some debris is in samples, too. As mites are very small - adults are 0.2 times 0.05 mm, white soft bodies, observing them using dissection microscope is very difficult, determination developmental stages and sex impossible.
Therefore, we consider to purchase an inverted microscope but we are not sure if to ask for that one which is equipped with relief contrasting method. If yes, which would be the best? Hoffman modulation contrast usually needs special condenser and lenses and is therefore more expensive. Zeiss microscope has so called Variable relief (VAREL) contrast which allows unilateral darkfield, perhaps convenient for our white mites.
As we do not have a possibility to try inverted microscope with relief contrast, I would very appreciate your opinion/advice.
Dear Colleagues: Tungsten bulbs change as they age (e.g., move toward more yellow wavelengths). Can anyone tell me if the mercury bulbs used in fluorescence microscopes also change with age? If so, is it possible that an old bulb might result in weakened fluorescence? --Many thanks, Jan Factor
--------------------------------------- Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Division of Natural Sciences Purchase College State University of New York Purchase, NY 10577 --------------------------------------- Ofc. Tel: 914-251-6659 Ofc. Fax: 914-251-6635 E-mail: jfactor-at-purvid.ns.purchase.edu ---------------------------------------
Whether your specialty involves fundamental research or practical applications of surface treatments and coatings, we encourage you to make a technical presentation at the 2nd ASM International Surface Engineering Congress & Exposition, to be held Sept. 15-18, 2003 in Indianapolis.
This premier event uniquely focuses on the practical engineering as well as the science of surface treatments and coatings. It's your best opportunity for networking with industrial engineers and scientists, industrial managers, and academic and government researchers.
To further enhance the technological and economic relevance of this year's technical sessions, our Congress will be co-located with America's largest heat treating event, the 22nd ASM Heat Treating Society Conference & Exposition, as well as the annual meeting of the North American Die Casters Association.
Abstracts are due by March 10 for presentations (platform and poster sessions) in the following areas:
Corrosion resistant surfaces PVD/CVD processes and applications Laser processes Thermal barrier coatings Computer modeling of deposition processes and applications Characterization of coatings and thin films Surface finishing effects and technologies Surface metrology Tribological coatings Residual stresses development and effect on properties Biomedical applications of coatings New deposition equipments and manufacturing Thermally sprayed coatings and applications Plasma enhanced processes Scanning probe microscopy for nanosciences
Please note that while "paper" submissions are encouraged for publishing in our proceedings volume, they are NOT required. To submit your abstract by March 10, please click on the following link: http://www.asminternational.org/absubmit_surface.cfm
Sincerely,
Dr. Oludele Popoola, General Chair Consultant Novi, MI popoola1-at-yahoo.com
Dr. Sudipta Seal, Program Chair University of Central Florida Orlando, FL sseal-at-pegasus.cc.ucf.edu
Dr. Arnold Deutchman, Expositions Chair Worthington BeamAlloy Corp., a Division of Worthington Industries, Inc. Columbus, OH ahdeutch-at-worthingtonindustries.com
Professor Narendra Dahotre, Proceedings Editor University of Tennessee-Center for Laser Applications Knoxville, TN ndahotre-at-utk.edu
Ask the vendor to bring the instrument you are considering to your lab for a demonstration. We always do this for expensive items. Any vendor who will not comply is not worth dealing with.
Geoff
Rostislav Zemek 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 users of ListServer, } } I kindly ask you for your advice concerning inverted microscope. We need } to count tiny mites in alcohol samples. Mites were washed from plants, so } some debris is in samples, too. As mites are very small - adults are 0.2 } times 0.05 mm, white soft bodies, observing them using dissection } microscope is very difficult, determination developmental stages and sex } impossible. } } Therefore, we consider to purchase an inverted microscope but we are not } sure if to ask for that one which is equipped with relief contrasting } method. If yes, which would be the best? Hoffman modulation contrast } usually needs special condenser and lenses and is therefore more } expensive. Zeiss microscope has so called Variable relief (VAREL) contrast } which allows unilateral darkfield, perhaps convenient for our white } mites. } } As we do not have a possibility to try inverted microscope with relief } contrast, I would very appreciate your opinion/advice. } } Thank you. } } Rostislav Zemek } } ------------------------------------------------------------------------------ } Rostislav Zemek Phone : 00420-387775227 } Institute of Entomology } Branisovska 31 Fax : 00420-385310354 } 370 05 Ceske Budejovice } Czech Republic E-mail: rosta-at-acarus.entu.cas.cz } ------------------------------------------------------------------------------ }
-- ********************************************** 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 mcauliff-at-umdnj.edu **********************************************
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Eighth Annual INTERNATIONAL 12-Day Short Course on
3D Microscopy of Living Cells June 15 - 26, 2003 (Pre-course: afternoon, June 14)
Seventh, Post-course Workshop on
3D Image Processing, June 28 - July 30, 2003
http:// www.3dcourse.ubc.ca/
Organized by Prof. James Pawley, (University of Wisconsin-Madison)
in association with
The Departments of Pharmacology and Physiology and the Brain Research Centre, University of British Columbia Vancouver, BC, Canada
DATES
Applications must be received by March 15, 2003 Deposit due April 15, 2003 Registration 5:00 - 7:00 PM Saturday, June 14, 2003 First Lecture 7:30 PM Saturday, June 14, 2003 Live-cell Course ends, noon Thursday, June 26, 2003 3D Image Processing Course, June 28 - 30, 2003
APPLICATIONS
Applicants must complete a questionnaire on the web to assess knowledge level, field of interest and proposed personal, live-cell, project. www.3dcourse.ubc.ca/application.htm
Enrollment will be limited to about 24-32 participants (exact number depends on number of 3D Systems available). Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with access to basic texts to read before the course begins. Application forms can be obtained from:
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-WISC.EDU
Additional information is available from: www.3dcourse.ubc.ca/brochure.htm
We expect to have at least 11, 3D microscope workstations for student use during 14 3D-Lab session and there will be an international faculty of 20.
Application deadlines:
Application forms must be received for screening by March 15, 2003. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2003. In general, refunds of the deposit will only be possible if someone on the waiting list can take the place. The remainder is due before Registration.
Pre-Course Tuition (1/2 day Basic Optical principles) $120 (US) 3D Live-cell Course Tuition (includes lunches and snacks): $2450 (US) 3D Image Processing Workshop Tuition (includes lunches and snacks): $900 (US)
Room/board about $40/day (US) depending on room type.
I hope that this includes all of the information that you need, but if not, please get back to me.
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-WISC.EDU 3D Microscopy of Living Cells Course, June 14 - 26, 2003, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, 2003
I'm in the market for a new critical point dryer, and trying to get a feel for the pros and cons of the various units that are available. If you have purchased a CPD in the last few years, would you please contact me (off-line)? I would appreciate hearing what you have and how well it works.
Thanks!
Leslie Eibest SEM lab manger Dept. of Biology Duke University
JiaMing manufacturing company, a world class forged iron, stained class, wood furniture, bamboo floor, stone product manufacturing company. Currently, is looking for European distributor. Please let us know if you're interested. You can also visit our web site www.jmwired.com
The article was by J. DeMay from Janssen Pharm in Beerse, Belgium and it's on pg 54 of EMSA Bulletin 14, 1 Spring 1984. It has a detailed appendix on how to make colloidal gold. The article is an extensive work with a zillion references.
I could either a.) photocopy the article and send it to Bill, b.) scan it in to PDFs and post their availability on the listserver, or c.) seek permission to reprint a shorter version in Microscopy Today. That said, Ed's suggestion to just go out and buy the stuff is a good one.
What does the community want?
Ron Anderson, Editor Microscopy Today and Editor of the EMSA Bulletin in 1984
-----Original Message----- } From: Edward Calomeni [mailto:calomeni-1-at-medctr.osu.edu] Sent: Tuesday, March 04, 2003 7:48 AM To: Microscopy-at-sparc5.microscopy.com
Morning Bill,
Why make your own gold, so much easier to buy and store premade?
Anyway, way back when there was a review article in the EMSA Bulletin entitled Colloidal Gold as a Marker and Tracer in Light and Electron Microscopy. Published around 1986??? Author was J. Dewey This is a nice review article on most aspects of colloidal gold labeling..
Other references to look up are:
1). Mulpferdt, H. Experientia, 1982 (38) pg 1127-1128 2). Roth, J. 1983b Techniques in Immunocytochemistry, Vol. II (Academic Press) pg 217-284.
best of luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
Looking for some lab in the Dallas area ( will consider other areas ) that could take some silicate powder material and heat it to 1100-1200C in a vacuum. Want to melt the material into a sample that can be sectioned and polished for microprobe analysis.
Thanks
Roy Beavers Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, Tx. 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
Greetings, Arc lamps probably do not change in spectrum as they age, but for sure they lose intensity. Sometimes when you change an old one, you will see a cloud of metalic deposit on the glass, presumably lowering the output intensity. In my experience the loss depends on the individual bulb, some lose more than others. But I have bulbs that are clearly and obviously less bright at the end of their life. Much rejoicing when the bulb gets changed.
Hope this helps, Tobias Baskin
} } } Dear Colleagues: Tungsten bulbs change as they age (e.g., move toward } more yellow wavelengths). Can anyone tell me if the mercury bulbs used } in fluorescence microscopes also change with age? If so, is it possible } that an old bulb might result in weakened fluorescence? } --Many thanks, Jan Factor } } --------------------------------------- } Jan Robert Factor, Ph.D. } Professor of Biology } --------------------------------------- } Division of Natural Sciences } Purchase College } State University of New York } Purchase, NY 10577 } --------------------------------------- } Ofc. Tel: 914-251-6659 } Ofc. Fax: 914-251-6635 } E-mail: jfactor-at-purvid.ns.purchase.edu } ---------------------------------------
Dear List, Thank you for the quick and informative responses. I have two protocols in hand and several promising references. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Jan I think mercury lamp may emit less light over the years for so many reasons (decreasing pressure or just dust cover). I would suspect more the filters: they may fade or change the properties under extended exposure to the UV light. In general, all optical components with age became dirtier and UV light may just make case the worse. Finally you may see funny sometimes colored layers of decomposed (or what?) material covered optical surfaces. It definitely will not increase the amount of light you have. Sergey
At 08:33 AM 3/4/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I can appreciate that anyone would prefer to have a vendor deliver an instrument to their door to do an on-site demonstration, but you also must realize that it is simply not possible in every instance. To say that "Any vendor who will not comply is not worth dealing with" is quite simplistic and offensive. I have nothing to do with this particular customer, but I can certainly understand how several excellent microscopy vendors may have difficulty justifying an on-site demo in the Czech Republic. I personally have been to the Czech Republic several times to demonstrate our systems, however, it can certainly not be justified in every instance. To dismiss vendors so capriciously would be doing a disservice to Dr. Zemek. Perhaps the company offering the best solution at the best price has a better method of demonstrating its performance than through an on-site demo. Technology these days is pretty impressive.
Speaking as a family business owner and microscopy vendor for nearly 40 years, I guess I take it a bit personally when vendors are treated with such disrespect. Scientists and vendors working together rather than as adversaries is much more productive and serves the scientific community much more effectively.
Best regards-
David
Geoff McAuliffe 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 } -----------------------------------------------------------------------. } } Ask the vendor to bring the instrument you are considering to your lab for a } demonstration. We always do this for expensive items. Any vendor who will not } comply is not worth dealing with. } } Geoff } } Rostislav Zemek 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 users of ListServer, } } } } I kindly ask you for your advice concerning inverted microscope. We need } } to count tiny mites in alcohol samples. Mites were washed from plants, so } } some debris is in samples, too. As mites are very small - adults are 0.2 } } times 0.05 mm, white soft bodies, observing them using dissection } } microscope is very difficult, determination developmental stages and sex } } impossible. } } } } Therefore, we consider to purchase an inverted microscope but we are not } } sure if to ask for that one which is equipped with relief contrasting } } method. If yes, which would be the best? Hoffman modulation contrast } } usually needs special condenser and lenses and is therefore more } } expensive. Zeiss microscope has so called Variable relief (VAREL) contrast } } which allows unilateral darkfield, perhaps convenient for our white } } mites. } } } } As we do not have a possibility to try inverted microscope with relief } } contrast, I would very appreciate your opinion/advice. } } } } Thank you. } } } } Rostislav Zemek } } } } ------------------------------------------------------------------------------ } } Rostislav Zemek Phone : 00420-387775227 } } Institute of Entomology } } Branisovska 31 Fax : 00420-385310354 } } 370 05 Ceske Budejovice } } Czech Republic E-mail: rosta-at-acarus.entu.cas.cz } } ------------------------------------------------------------------------------ } } } } -- } ********************************************** } 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 } mcauliff-at-umdnj.edu } **********************************************
-- David Henriks Vice President
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Can anyone tell me if the mercury bulbs used } in fluorescence microscopes also change with age? If so, is it possible } that an old bulb might result in weakened fluorescence? } --Many thanks, Jan Factor
Mercury lamps lose their output as a function of time. At the end of the rated life of the lamp the output is diminished by 30%, per the manufacturer. Not letting the lamp warm up before shutting it off can also cause the lamp to lose output, by darkening the envelope. It may also ruin the lamp completely. These lamps must be on for a minimum of 15 minutes after being lit, per the manufacturer. I suggest 20 minutes as a rule.
Alignment of the lamp is critical and can have a dramatic effect on the light delivered to the specimen. It is very common for the lamp to be installed incorrectly, or to have someone try and "adjust" the lamp for you and throw the alignment off. The adjusting mechanism in the lamphouse of many of these systems freezes up over time as the lubricants are dried up by the intense heat of the lamp and this also contributes to an inability to make the proper adjustments...
Good luck,
Try Lamp Technology or Bulbman as a resource for replacement lamps. Great prices, good service.
Std disclaimer, I have purchased thousands of lamps from both but have no other relationship with them.
David Burton Optical Specialist, University of Washington dbuw-at-u.washington.edu
We might be in the market for a new digital camera for microscopy/image analysis. I am currently using a Nikon DXM 1200 (12 magapixels) but wonder what else is out there as the development of such devices is going so quickly.
Contacts from users with likes/dislikes and also from the trade would be welcome.
Obs/NB New postal/visiting address from July 2002!
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
We are trying to make small optical cells for use under the optical microsccope, by building up square glass "walls" on a microscope slide, or mounting a metal washer on the same. The cavity is to contain some rather agressive solvent mixtures, also sometimes to be heated, so sticking with superglue or Aralidite does not always make a sound cell. Could anyone tell us of a tough adhesive for these purposes?
(School essay, AD3000: "The greatest playwrite of the Early Computer Age was William Shakespeare, who wrote the Comedy of Errors.")
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
Ron et al. I vote for creating Protocol Bank on the MSA WWW site and having Debby's and Jan's protocols there as pdf files. Marek.
At 03:17 PM 3/4/03 -0500 Tuesday, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I went through all the SEM tips in the archives but I still couldn't find answers to two specific problems that I am facing:
1. Mouse Embryos: After OTO processing and CPD the samples became very fragile and some are even damaged during the run. How does one get the embryos onto the stub and orient them without damaging them? Or should the sample be attached on to something before CPD?
2. Insects: In preparing whole insects for SEM, the problem I found was that after CPD they also become very brittle, easily damaged. Some suggest to stick a pin through the insect, but I don't know at what stage should this be done. Before fixation? before CPD?
Eleana Sphicas Bio-imaging Resource Center Rockefeller university (212)-327-8125
} Geoff: } } I can appreciate that anyone would prefer to have a vendor deliver an } instrument to their door to do an on-site demonstration, but you also } must realize that it is simply not possible in every instance.
That may be true.
} To say } that "Any vendor who will not comply is not worth dealing with" is quite } simplistic and offensive.
Hey, times are very tight here in University research land. We spend our money the way we want to. We can't afford to buy equipment we have not tested. If this offends you, so be it. In 30 years in university science, I have never had a vendor refuse to demo a product, even an inexpensive one.
} I have nothing to do with this particular } customer, but I can certainly understand how several excellent } microscopy vendors may have difficulty justifying an on-site demo in the } Czech Republic.
If that is the nature of their business, then that is the nature of their business. The vendor runs his business the way he wants to, the customer spends his money the way he wants to. Capitalism.
} I personally have been to the Czech Republic several } times to demonstrate our systems, however, it can certainly not be } justified in every instance. To dismiss vendors so capriciously would } be doing a disservice to Dr. Zemek.
No, Dr Zemek can make his own decisions. I was suggesting one course of action for his consideration.
} Perhaps the company offering the } best solution at the best price has a better method of demonstrating its } performance than through an on-site demo. Technology these days is } pretty impressive.Speaking as a family business owner and microscopy vendor for } nearly 40
} years, I guess I take it a bit personally when vendors are treated with } such disrespect.
Not disrespect, it is just the way things operate in my world. How is 'not buying a product without a demo to see if it fits your lab's needs' disrespectful??
} Scientists and vendors working together rather than as } adversaries is much more productive and serves the scientific community } much more effectively.
That is a very nice, general statement that no one would argue with. It is also 180 degrees opposite of buying a product without knowing if it will solve your problem.
Geoff
} Best regards- } } David } } Geoff McAuliffe 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 } } -----------------------------------------------------------------------. } } } } Ask the vendor to bring the instrument you are considering to your lab for a } } demonstration. We always do this for expensive items. Any vendor who will not } } comply is not worth dealing with. } } } } Geoff } } } } Rostislav Zemek 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 users of ListServer, } } } } } } I kindly ask you for your advice concerning inverted microscope. We need } } } to count tiny mites in alcohol samples. Mites were washed from plants, so } } } some debris is in samples, too. As mites are very small - adults are 0.2 } } } times 0.05 mm, white soft bodies, observing them using dissection } } } microscope is very difficult, determination developmental stages and sex } } } impossible. } } } } } } Therefore, we consider to purchase an inverted microscope but we are not } } } sure if to ask for that one which is equipped with relief contrasting } } } method. If yes, which would be the best? Hoffman modulation contrast } } } usually needs special condenser and lenses and is therefore more } } } expensive. Zeiss microscope has so called Variable relief (VAREL) contrast } } } which allows unilateral darkfield, perhaps convenient for our white } } } mites. } } } } } } As we do not have a possibility to try inverted microscope with relief } } } contrast, I would very appreciate your opinion/advice. } } } } } } Thank you. } } } } } } Rostislav Zemek } } } } } } ------------------------------------------------------------------------------ } } } Rostislav Zemek Phone : 00420-387775227 } } } Institute of Entomology } } } Branisovska 31 Fax : 00420-385310354 } } } 370 05 Ceske Budejovice } } } Czech Republic E-mail: rosta-at-acarus.entu.cas.cz } } } ------------------------------------------------------------------------------ } } } } } } } -- } } ********************************************** } } 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 } } mcauliff-at-umdnj.edu } } ********************************************** } } -- } David Henriks } Vice President } } South Bay Technology, Inc. } 1120 Via Callejon } San Clemente, CA 92673 USA } } TEL: +1-949-492-2600 } Toll-free in the USA: +1-800-728-2233 } FAX: +1-949-492-1499 } } email: henriks-at-southbaytech.com } } Celebrating 38 years of providing Materials Processing Solutions for } Metallogaphy, } Crystallography and Electron Microscopy. } } Please visit us online at www.southbaytech.com. } } The information contained in this message and any attachments is } privileged and } confidential. This message is intended for the individual or entity } addressed. } If you are not the intended recipient, please do not read, copy or } disclose this } communication. Notify the sender of the mistake by calling } +1-949-492-2600 and } delete this message from your system.
-- ********************************************** 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 mcauliff-at-umdnj.edu **********************************************
Hello everyone, I've been trying to enlarge a "virtual museum" of older, significant scopes and equipment used in electron microscopy. If you have a picture of older equipment (out of use or still being used) I would like to include it on the site. I would also like to include some information about the equipment (dates, usage, location, any interesting features, etc..) and cite the provider of the image as a contributor. This could also be a link to an existing image at your site. At present it represents equipment collected by the late Dr. Jerry Paulin. I would like to add a "Contributed Collection" addition to this. I think it would be great to show EM students the variety and provide them an appreciation for the changes in equipment.
The museum, as it stands now, can be seen at www.uga.edu/caur/museum.htm
Thanks in advance for your participation. John Shields EM Lab University of Georgia Athens, GA 30602 jshields-at-cb.uga.edu
What I've done for such delicate samples as you describe and with some success is to take a pin (like an insect mounting pin, or push pin) cut to about 1/4 inch in length, file a flat end, dip end in colloidal carbon paint, gently touch that end to "underside" (or side opposite the one you want to look at) of insect or embryo. The sample will stick to the paint so then you can lift it up. Hold the pin by its side with a clamping type fine tipped forceps.
Next step is to mount it onto an SEM sample stub. You can drill a small hole into the stub surface that will take the diameter of the pin, or use a clamping type SEM stub, both of which I have here. After mounting/clamping the pin onto the stub, paint carbon paint over the stub surface, then you will have a nice dark background behind the sample, which may also be conveniently out of focus, due to height of sample above stub surface, which helps to give dark featureless background for those full body glamour shots!
Good luck!
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Bera
} Hi list members, } } I went through all the SEM tips in the archives but I still couldn't find } answers to two specific } problems that I am facing: } } 1. Mouse Embryos: After OTO processing and CPD the samples became very } fragile and some are even damaged during the run. How does one get the } embryos onto the stub and orient them without damaging them? Or should the } sample be attached on to something before CPD? } } 2. Insects: In preparing whole insects for SEM, the problem I found was } that after CPD they also become very brittle, easily damaged. Some suggest } to stick a pin through the insect, but I don't know at what stage should } this be done. Before fixation? before CPD? } } Eleana Sphicas } Bio-imaging Resource Center } Rockefeller university } (212)-327-8125
A lot of the problem with arc sources is the electrodes. As the arc files over a long period of time, the electrodes erode, decreasing intensity. This problem adds to the deposition you have mentioned.
OptiQuip has a new stabilizer (LTS) and power supply (model 1600) for their Nobska fluorescence illuminators which automatically tests the lamp intensity 2000 times/s, compares it to a stored intensity, then signals the power supply to adjust the voltage to maintain the intensity. Normal stabilized power supplies control just the flicker. It's really great for everything from measurement to montage to deconvolution.
We ran an article last November in American Lab which discusses this new approach in detail: Foster, B. "Optimizing illumination for Microscopy and Measurement", Am Lab, Nov 2002, Pp13-17.
Caveat: MME has no financial interest in this product.
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
"Why didn't they teach us that sooner?" ... probably because no one thought to call MME about customized, on-site courses. Offered in all areas of microscopy, sample prep,and image analysis, they make an immediate impact on your productivity. -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 03:32 PM 3/4/03 -0600, Tobias Baskin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I use a 5/0 Red Sable artist's brush. You can buy these from the EM suppliers like Ted Pella for a couple bucks. I use them on embryos and insects, just like you. Just gently touch the sample and it will usually stick to the hairs. Then touch it down to your adhesive substrate. If the sample won't stick to the brush, touch the brush to your tongue (or touch your finger to your tongue then touch the brush to your finger). Saliva is a wonderful thing.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
At 09:26 AM 3/5/2003 -0500, Eleana Sphicas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Silicone sealant will probably work(something like Silastic (TM)). It's both heat and solvent resistant. You can try the hardware store variety, or contact Dow, GE, or one of the other manufacturers if you need something more specialized.
On Wednesday, Mar 5, 2003, at 05:38 America/New_York, Robert H. Olley 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 trying to make small optical cells for use under the optical } microsccope, by building up square glass "walls" on a microscope slide, } or mounting a metal washer on the same. The cavity is to contain some } rather agressive solvent mixtures, also sometimes to be heated, so } sticking with superglue or Aralidite does not always make a sound cell. } Could anyone tell us of a tough adhesive for these purposes? } } (School essay, AD3000: "The greatest playwrite of the Early Computer } Age was William Shakespeare, who wrote the Comedy of Errors.") } } +-----------------------------------------+ } Robert H.Olley } J.J.Thomson Physical Laboratory } University of Reading } Whiteknights } Reading RG6 6AF } England } +-----------------------------------------+ } Phone: } {direct line +44 (0) 118 9318572 } {University internal extension 7867 } Fax: +44 (0) 118 9750203 } Email: R.H.Olley-at-reading.ac.uk } URL: http://www.reading.ac.uk/~spsolley } +-----------------------------------------+ } } } } All the best, Andy Buechele
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } Eleana Sphicas {sphicae-at-rockefeller.edu} 03/05/03 09:26AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi list members,
I went through all the SEM tips in the archives but I still couldn't find answers to two specific problems that I am facing:
1. Mouse Embryos: After OTO processing and CPD the samples became very fragile and some are even damaged during the run. How does one get the embryos onto the stub and orient them without damaging them? Or should the
sample be attached on to something before CPD?
Fixed and dehydrated tissues will become brittle. Process them adhered to something. Amazingly SuperGlue does a nice job on fresh tissue. Use very sparingly and do not cover or the fumes will deposit on the surfaces. There are also ways to bind to glass coverslips (aminosilane), and then just process the coverslip itself.
2. Insects: In preparing whole insects for SEM, the problem I found was that after CPD they also become very brittle, easily damaged. Some suggest
to stick a pin through the insect, but I don't know at what stage should this be done. Before fixation? before CPD?
Ha, the bane of my recent existence. How to keep insects looking lively. Already dried specimens may be placed in a steam bath to absorb moisture and then CAREFULLY arranged (Some of my researchers just plunk in warm water overnight with a little surfactant). I prefer the steam because it doesn't require CPD or HMDS so long as you control it. If samples are in alcohol or fixative , you may pin mount them, but not through the body. Splay out the critter, and place a pin to either side of the body, angled so that they cross just over the structure. It takes time and lots of practice especially to pin down all the legs, antenna, wings, body... but the results are well worth it. I use silicone mats or filter paper underneath. If you don't have the mini pins, a cactus works really well (my preference). From this point dry in whatever manner suits you.
Fresh samples are most easily done by getting them to land on a damp piece of filter paper. When happy, set on a liquid nitrogen chilled block to snap freeze. I have also had success getting creepers to walk across tape and then freezing in a lab freezer to kill. Freeze drying would be the preferred method from this point.
Method three- Use an ESEM or cryo stage equipped instrument. Getting them to sit still under the beam is the major drawback with ESEM
Good luck!!
Eleana Sphicas Bio-imaging Resource Center Rockefeller university (212)-327-8125
Someone just dropped off a handful of "ancient" coins and wants to know if there is anything an SEM could tell him about their authenticity. The person dropping them off is not a crackpot treasure hunter type. He is a careful and knowledgeable professor who is really stumped.
They come from Syria, where they were bought from a street vendor. They are supposed to be silver and appear to be the real thing. If fake, they are really good fakes. If I got the story right, the vendor did not claim that they were authentic, but couildn't, wouldn't say where they came from. The price was about right for reproductions, but on closer inspection, they do not look like obvious forgeries.
We suspect they may be cast rather than struck and were wondering if there are any characteristic features that would tell us how the coins were made. We will do a little EDS to check on the metal, are there any trace metals that might be a give away, either way?
If you know anything about coins or anyone who we could contact, we would appreciate it very much if you could help us out.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
On Tuesday, March 4, 2003, at 12:17 PM, MicroscopyToday wrote:
} Way to go Ed--you've got a great memory! } } The article was by J. DeMay from Janssen Pharm in Beerse, Belgium and } it's on pg 54 of EMSA Bulletin 14, 1 Spring 1984. It has a detailed } appendix on how to make colloidal gold. The article is an extensive } work } with a zillion references. } } I could either a.) photocopy the article and send it to Bill, b.) scan } it in to PDFs and post their availability on the listserver, or c.) } seek } permission to reprint a shorter version in Microscopy Today. That said, } Ed's suggestion to just go out and buy the stuff is a good one. } } What does the community want? } Dear Ron, I can print out the article from PDF files, so that would be my preference; however, either of the other options is acceptable. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Jonathan: I've had a fair amount of experience using the SEM in the authentication of ancient coins. Based on your description of the origin of these coins from a street vendor, the chances of their being modern fakes are about 99.99% The individuals in the countries where ancient coins are found know exactly what genuine ancient coins are worth, especially silver ones, and they know where to sell them, which is not a vendor on a street corner.
You are correct in your assumption that they are most likely a cast fakes; most inexpensive reproductions are. Look carefully for a joining line at the edge where the two halves were glued together, or look for an edge that has been filed and then roughened to hide the joining line. Also, look for small casting bubbles in the metal.
In terms of the SEM, do quantitative analysis of the metal. If the coin is covered with toning, you may have to scrap a minute area clean to get an accurate analysis. If the metal is too pure, i.e. greater that 92% silver, be very suspicious. Look for trace metals. In genuine coins you may see small amounts of lead (~2 wt%), gold (less than 1 wt% and often below minimum detectable concentration), as well as iron. If it is cast, you are more likely to see some common casting metal alloy that just looks like silver. Look at the toning using EDS. Silver fakes are often toned with paint (titanium), iodine, etc. Genuine toning will usually show lead, iron, or chlorine from silver chloride (horn silver as it is know to ancient numismatists). If the composition of the toning is identical all over the coin, then be suspicious. Genuine ancients usually have considerable variation in chemical composition of the toning.
I hope this helps. Stanley L. Flegler, Director Center for Advanced Microscopy Michigan State University
I could put you in touch with one of the authors of that study, if you wish to pursue it further.
"To achieve results never before accomplished, we must employ methods never before attempted." -Sir Francis Bacon
Jeffrey A Fortner Argonne National Laboratory CMT/205 9700 S. Cass Avenue Argonne, IL 60439 (630) 252-5594 FAX: (630) 972-4438
} ---------- } From: jmkrupp-at-cats.ucsc.edu } Sent: Wednesday, March 5, 2003 1:40 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM (?) of ancient coins } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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: } } Someone just dropped off a handful of "ancient" coins and wants to know if } there is anything an SEM could tell him about their authenticity. The } person dropping them off is not a crackpot treasure hunter type. He is a } careful and knowledgeable professor who is really stumped. } } They come from Syria, where they were bought from a street vendor. They } are } supposed to be silver and appear to be the real thing. If fake, they are } really good fakes. If I got the story right, the vendor did not claim that } they were authentic, but couildn't, wouldn't say where they came from. The } price was about right for reproductions, but on closer inspection, they do } not look like obvious forgeries. } } We suspect they may be cast rather than struck and were wondering if there } are any characteristic features that would tell us how the coins were } made. } We will do a little EDS to check on the metal, are there any trace metals } that might be a give away, either way? } } If you know anything about coins or anyone who we could contact, we would } appreciate it very much if you could help us out. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
Barbara, we are using two sets of OptiQuip 1600 Power supply and LTS, which runs two of our Mercury lamp housings. We have shipped (Currently packing one) 7 times as of today in the year starting from January 1, 2002 (excluding this time) to New York for repair. That means an average of repair on either one of the system in every two months. Most of the time the Power supply blows the main fuses and burns the cord, which is connecting the LTS. It will take a two weeks to get the system back. If you have used this system for a long time, do you had any problems like these? We appreciate your comments.
We have now asked the company's President to put a full stop on this saga through our local Olympus Representative Mr. David Kinast. Thanks
Shiv
Mayandi Sivaguru, PhD Associate Director Molecular Cytology Core Facility 2, Tucker hall University of Missouri Columbia, MO 65211-7400
Voice: 1-573-882-4895 Fax: 1-573-882-0123
www.biotech.missouri.edu/mcc/
At 11:42 AM 3/5/03 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
There is a large body of information on this in the literature of the fields of archaeometry and art conservation science. The criteria that are important depend on the culture represented by the coins, their age, the burial conditions, ore types exploited for the metals and the treatment or mishandling that they have received since they were found.
Compositional information is often not sufficient by itself to make a determination. Phase analysis (by X-ray diffraction) or microstructural analysis by very limited metallographic testing (to minimize destructive action)may be required.
I've never heard this term "toning" before. Perhaps you are referring to the corrosion layer, oxide or patina that metal surfaces typically acquire and that may be falsified in the case of a forgery.
Unfortunately the criteria that you site could be highly misleading to someone unfamiliar with numismatics or with the methods that have been used over the years to treat ancient metals. Applying these criteria in the way that you suggest could result in a large proportion of legitimate ancient pieces being rejected.
On the other hand (other side of the coin?) in antiquity there were often large numbers of counterfeit coins in circulation. This is particularly true in an area like Syria that was a cross roads for many cultures and often outside close control of distant capitols. These ancient counterfeits are often debased but extremely valuable historical indicators of conditions at the time. Your criteria seem to assume that to be an antiquity a coin must be of high quality and from the official mint.
I would urge you to run any coins of potential value by a knowledgable numismatist before engaging in destructive tests or attempting to clean them. And not to base a decision on broad assumptions such as those regarding the purity of the metal.
John Twilley Art Conservation Scientist
-------Original Message------- } From: "Stanley L. Flegler" {flegler-at-pilot.msu.edu} Sent: 03/05/03 03:38 PM To: Microscopy-at-sparc5.microscopy.com, Jon Krupp {jmkrupp-at-cats.ucsc.edu}
Jonathan: I've had a fair amount of experience using the SEM in the authentication of ancient coins. Based on your description of the origin of these coins from a street vendor, the chances of their being modern fakes are about 99.99% The individuals in the countries where ancient coins are found know exactly what genuine ancient coins are worth, especially silver ones, and they know where to sell them, which is not a vendor on a street corner.
You are correct in your assumption that they are most likely a cast fakes; most inexpensive reproductions are. Look carefully for a joining line at the edge where the two halves were glued together, or look for an edge that has been filed and then roughened to hide the joining line. Also, look for small casting bubbles in the metal.
In terms of the SEM, do quantitative analysis of the metal. If the coin is covered with toning, you may have to scrap a minute area clean to get an accurate analysis. If the metal is too pure, i.e. greater that 92% silver, be very suspicious. Look for trace metals. In genuine coins you may see small amounts of lead (~2 wt%), gold (less than 1 wt% and often below minimum detectable concentration), as well as iron. If it is cast, you are more likely to see some common casting metal alloy that just looks like silver. Look at the toning using EDS. Silver fakes are often toned with paint (titanium), iodine, etc. Genuine toning will usually show lead, iron, or chlorine from silver chloride (horn silver as it is know to ancient numismatists). If the composition of the toning is identical all over the coin, then be suspicious. Genuine ancients usually have considerable variation in chemical composition of the toning.
I hope this helps. Stanley L. Flegler, Director Center for Advanced Microscopy Michigan State University
I'm not sure if your cells would be compatible, but we often used a non-CO2 dependent HEPES buffer for relatively long term atomic force microscopy/calcium imaging/electrophysiology experiments with primary hippocampal neuron and glial cultures. Might be worth looking at.
All the best,
Raj
Dr. Raj Lartius Novascan Technologies, Inc. Iowa State University Research Park 2501 North Loop Drive Ames, Iowa 50010 USA
At 01:14 PM 2/25/03, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Novascan Technologies, Inc. Iowa State University Research Park 2501 North Loop Drive Ames, Iowa 50010 USA
is anyone having any information on the lens design of the old ISI SMS 2 SEM. I'd require data on the geometry of the pole pieces as well as the number of A-t. But for the meantime typical values for the lens currents might already be helpful.
Besides the replies already given, I have used "live insect handling forceps" for both types of samples mentioned (and many other fragile, driec samples). These are blunt or fine-pointed forceps made of thin, flexible stainless steel that allow a sure grip with minimal force. As the name implies, they were originally developed by entomologists, but I believe not for live insects, but minute, dried insects in collections. I got mine from Fine Science Tools, cat #26029-10 & 26030-10 http://www.finescience.com/ . There should be other sources. The other trick is to put a pointy bit of latex rubber on the end of a dissecting needle and rub it on silk or wool or something to give it a charge. The static electricity can then be used to pick up and move fragile specimens. (My apologies to whoever first did this long ago, but I don't remember whom to credit.)
Phil
} Hi list members, } } I went through all the SEM tips in the archives but I still couldn't } find answers to two specific } problems that I am facing: } } 1. Mouse Embryos: After OTO processing and CPD the samples became } very fragile and some are even damaged during the run. How does one } get the embryos onto the stub and orient them without damaging them? } Or should the sample be attached on to something before CPD? } } 2. Insects: In preparing whole insects for SEM, the problem I found } was that after CPD they also become very brittle, easily damaged. } Some suggest to stick a pin through the insect, but I don't know at } what stage should this be done. Before fixation? before CPD? } } Eleana Sphicas } Bio-imaging Resource Center } Rockefeller university } (212)-327-8125
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Other than finding a joining line and/or casting bubbles, there is no one single factor that can be used. Instead look for "mistakes" that a modern counterfeiter may have made.
Doing an EDS check on the metal is an excellent first method of determining authenticity. A high proportion of these tourist fakes are cast and can be eliminated immediately by their alloys. A surprising number do not even contain silver, but instead a casting alloy such as Bismith, Lead, Tin, and Cadmium. A surprising number are cast using solder!
Among the struck (as opposed to cast) fakes, the single most common mistake of modern counterfeiters is to use either four nines pure silver or silver from common silver coins (for example 90% Ag, 10% Cu) without any trace metals. The ancients were not able to produce silver of ultra high purity and trace amounts of other metals (lead, iron, and gold) are present. Athens produced some of the highest silver content ancient coins, but even then, they are perhaps 92% to 94% silver with other trace elements. An excellent source of information on this area is {underline} Metallurgy in Numismatics {/underline} , vol 1 and 2, published by the Royal Numismatic Society, {underline} Analysis of Ancient Metals {/underline} , by E. Caley, and {underline} The New Style Silver Coinage of Athens {/underline} , by Margaret Thompson.
The term toning is often used by numismatists rather than corrosion layer, oxide, etc. It is possible that a genuine ancient coin has been cleaned and artificially retoned. Therefore, if you find titanium or iodine on the surface, it still could be ancient.
There were counterfeit coins in ancient times. The most common example is known as a "fourre." A copper core or blank was used with a thin layer of silver bonded to it. These were then struck using dies that attempted to copy the genuine coins in circulation. In ancient times, silver coins were often tested by making a test cut on the edge to see if a copper core was present. A number of these "tested" coins have survived and are in collections, etc. The ancient counterfeiter made a profit by passing these coins off as high silver coins, when in fact they were mainly copper. Most of these coins can be identified by the breaks in the silver layer that developed over the centuries. If there is any doubt, the coin can be placed in the SEM and if a suspicious area proves to copper, then that is a good indication that it is a fourre. If further proof is needed, specific density testing can be done, short of total destructive analysis.
Recently, we just installed a new JEOL 2010F TEM. The TEM has a 2k*2K ultrascan CCD (not yet installed), EDS, Enfina PEELs and Digiscan STEM system. We also have both Emispec and Gatan systems. This is a user based facility. The TEM is booked everyday.
We are trying to figure out a way for data storage, to let users save data for a period of time and let them transfer data through Internet.
I appreciate for the following informations:
what is the data storage system in your Lab? how much data in a period of time (a day, a week) the users save? how frequently they save and transfer the data?
Any other suggestion on internet security is also welcome
Thank you very much
Sincerely,
Jinguo Wang
Jinguo Wang, Ph.D The Pennsylvania State University Materials Research Institute 194 Materials Research Institute Building University Park, PA 16802 Tel: (814) 865-9285 Fax: (814) 863-8561, (814) 863-0637 email: jqw11-at-psu.edu
A histology question for the light microscopists out there: Are Haversian systems present in the flat bones formed by intramembranous ossification or are they only in long bones? Thanks, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I have the Cressinton MTM10 thickness monitor. It has stopped working. Does anyone know anything about this monitor and what I can check other than the fuse which seems to be fine? OR does anyone know how to get in touch with Cressington. I have left phone messages and sent them email from their web site and have gotten no replies.
Thanks in advance for any suggestions.
Mei Lie -- Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
I installed a Windows NT server 6 years ago. I made each microscope with a computer interface a client. There is a Philips TEM with Gatan CCD, Hitachi SEM, Leica confocal, 2 graphics workstations, and a backup server. All the systems run Windows based software so they can be networked as clients of the server (we include one Mac workstation for poster printing). There is also a UNIX based deconvolution scope that shares some parts of the facility. The server has about 60 gig of RAID 5 storage. Images acquired from any client are saved directly to the server. Images are never saved to the client workstations so the workstations stay more or less as delivered. No extraneous software is written to the workstations so the microscope application that is installed at setup remains unchanged. Users get an account on the server and this allows them space on the server. Once the image is written to the server it can be downloaded on the LAN, on the Internet via HTTP, or via FTP. Even with the confocal on the network the 60 gig is plenty of space for now. The server is considered a "transfer station" for information. We do not store images. My policy is that images over 30 days old may be removed at any time without notification. In actual practice, I do not remove anything less than 6 months old. If you want to provide storage, you may need more space. We have over 200 accounts so we leave storage and archiving to the individual but we do provide CD writing and DVD writing hardware in the facility. We do not set quotas on data storage. That feature is not native to Windows NT4.0 but can be added from another vendor. That feature is available in Windows2000 Server. Storage is cheap so quotas are not needed here yet. Security is managed by keeping aware of patches and fixes to the operating system. A firewall is being considered. In the 6 years that the system has been running there has been no security breach and no viruses.
Contact me directly if you have questions.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
At 11:39 AM 3/6/2003 -0500, Jinguo Wang wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
We may need to buy a table to put our Zeiss Axiovert on. I am aware of the various vibration isolation systems discussed here previously, and have used a heavy slab on sorbothane feet on a standard table for years with good success. However the tables that we received with our new building seem to have alot of flex in teh legs, so that even with an isolated slab on top, we are concerned about stage vibration. Can anyone suggest a good solid table to use as a base, or should I just go for a TMC/Kinetic systems table. Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
We have been very happy with our Snap server otherwise known as an NAS server. Basically just a bunch of hard drives and ethernet card. I have the Quantum Snap with 240G of space mirrored. They are scalable and you can add a bunch together to really serve some data. Users access their data in a variety of ways, and it took about as long to set up as to unpack. It also serves as the backup dump for the lab PC's.
We serve roughly 70 researchers and their associates annually generating 15-20k images. Our Molecular group uses a bunch of these as well for their data sets (get big quick) and the users write data to CD's as they fill up their quotas. Again authentication is done on a separate server and it all sits behind the SI firewall keeping the riffraff out. Good luck
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } Jinguo Wang {jqw11-at-psu.edu} 03/06/03 11:39AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers
Recently, we just installed a new JEOL 2010F TEM. The TEM has a 2k*2K ultrascan CCD (not yet installed), EDS, Enfina PEELs and Digiscan STEM system. We also have both Emispec and Gatan systems. This is a user based facility. The TEM is booked everyday.
We are trying to figure out a way for data storage, to let users save data
for a period of time and let them transfer data through Internet.
I appreciate for the following informations:
what is the data storage system in your Lab? how much data in a period of time (a day, a week) the users save? how frequently they save and transfer the data?
Any other suggestion on internet security is also welcome
Thank you very much
Sincerely,
Jinguo Wang
Jinguo Wang, Ph.D The Pennsylvania State University Materials Research Institute 194 Materials Research Institute Building University Park, PA 16802 Tel: (814) 865-9285 Fax: (814) 863-8561, (814) 863-0637 email: jqw11-at-psu.edu
I am trying to get our kevex system running for fully quantitative analysis and have been saving reference standards. When I try to use the standards in deconvolution some are accepted while others are rejected with the message "not a standard for this element and line". Elements up to Al seem to be accepted while everything after Ca is rejected. We thought it may have something to do with the Kb line. I've tried everything I can think of, re-saving etc, I know I have used the same process for all the standards. Any suggestions?
Kind Regards,
Jodi Reynolds. Onesteel Wire Mills Newcastle Australia
Does anyone have any recommndations on laminar flow hoods for optical microscopy? We are looking for one to create a clean space for sample prep and optical micrscopy.
Joe Neilly, Research Investigator Abbott Laboratories R45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
Hi folks - does anyone have a confocal with a Coherent Enterprise II 651 UV argon laser or similar, operating close to high resolution TEMs and SEMs - are there likely to be any problems with electrical interference, either in the power supply or radiated?
cheers
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University, Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 http://www.anu.edu.au/EMU
Hi All, I'd like to hear opinions about people's impression of the OEM (original equipment manufacturer sponsored) service they receive for their TEM's. Is it timely (average time from request to completion)? Are issues resolved the first time (call backs for same problem)? Do you feel it is cost effective? Is your downtime excessive? Minimal? Service personnel to service area ratio? Feel free to add any topic I might have missed. We are looking at acquiring new instrumentation, and thus are trying to evaluate every aspect of such a purchase.
All responses will remain confidential.
Thanks in advance, Randy Nessler University of Iowa Central Microscopy Research Facility Iowa City, Iowa 52242 Phone 319-335-8142
Listers, An immediate opening for an experienced (T)EM tech is available here at the Carolinas Medical Center in Charlotte, NC. Please see the listing and apply on-line at {http://www.carolinashealthcare.org} www.carolinashealthcare.org.
Go to Find A Job, then Job Search. Complete Step 1 with "Research Services," Step 2 with "Cannon Research Center," leave Step 3 blank, and click Find (Step 4.) Page down to Res Tech II, Position number 74366.
After viewing the listing (shown below and at the MSA job website,) if interested go to "Apply On Line," and submit your cva/resume indicating the position number.
Hope to hear from you soon!
Res Tech II POSITION NUMBER: 74366 / LOCATION: Cannon Research Center / CATEGORY: Research Services FT,M-F,days. Prepare chemical reagent's, follow chemical protocols, fixation through embedding of specimen, photo darkroom techniques with print processing & related duties. Bio/Chem/Med laboratory, must be able to stand & walk for extended periods. BA/BS in Biology/ Chemistry /or related.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor Cannon Electron Microscopy Lab Carolinas Medical Center P.O. Box 32861; Charlotte, NC 28232-2861 Ship to: 1000 Blythe Blvd; Charlotte, NC 28203 Ofc:704-355-1267 ; Lab:704-355-7220 ; Fax 704-355-0589 E-mail: {mailto:WWiggins-at-Carolinas.org} Winston.Wiggins-at-CarolinasHealthCare.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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PLEASE CIRCULATE THIS TO ALL THAT MAY BE INTERESTED
The Midwest Microscopy and Microanalysis Society (MMMS) is co-sponsoring the following upcoming meetings:
March 27th at Northwestern University (with the Biological Imaging Facility atNorthwestern) Topics include Tomography, Cryo FESEM in addition to others (see details below)
Location: Norris University Center, Lewis Room, Evanston,IL
May 13th - Joint meeting with the Milwaukee ASM Chapter
Topic - Color Metallography presented by George F. Vander Voort, Buehler Ltd.
Location: Klemmer's Banquet Center,10401 W. Oklahoma Ave.Milwaukee, WI 53227(414) 541-0401, 5:00pm Social 6:00pm Dinner 7:00pm Program
For additional information on either program contact Robb Mierzwa (mierzwa-at-jeol.com) or Jim DiOrio (jim_diorio-at-baxter.com)
Northwestern Program: (3/27/03)
9:00 am : Registration, 9:45 Welcome and introduction
10:00 Brad Marsh,Univeristy of Colorado at Boulder:3D Structure studies of the pancreatic beta cell by high resolution EM tomography
11:00 Valerie Kilman,Northwestern University:A new role for an old kinase in the Drosophila circadian clock
11:30 Veronica Ledoux,Northwestern University:Serial EM study of inhibitory hippocampal synapses
12:00 Lunch
1:00 Stan Erlandsen,Univeristy of Minnesota Medical School:Visualizing the glycocalyx:correlative microscopy on cell interaction and detecting individual molecules with immunogold by cryoSEM
2:00 Kendrick Boardman,Northwestern University:Lymphangiogenesis:roles of interstitial fluid flow and fluid channel formation
2:30 Greg Beitel,Northwestern University:Unexpected roles for the Na K ATPase in epithelial tube-size control and septate (tight)junctions organization
3:00 Reception:
4:00 Close
Driving directions to Northwestern University Evanston Campus available on the Web. at http://www.northwestern.edu/campus/directions/
RSVP Required
Please send RSVP via email,phone,or fax to: Robb Mierzwa,MMMS Program Coordinator c/o JEOL USA,Inc. 3906 Lisa Avenue Sheboygan,WI 53083 phone:(920)803-8945;FAX:(920)803-8946 email:mierzwa-at-jeol.com Admission:Free to MMMS Members MMMS Membership:$10.00 MMMS membership available at registration
Mei Lie, of course we've already contacted you, but for clarification to the rest of the list: Ted Pella, Inc. provides sales & service for Cressington vacuum equipment (except the CFE-60) in North, Central and South America. You can reach us at (800) 237-3526.
Tom Pella Ted Pella, Inc. http://www.tedpella.com
-----Original Message----- } From: Mei Lie Wong [mailto:wong-at-msg.ucsf.edu] Sent: Thursday, March 06, 2003 10:23 AM To: Microscopy-at-sparc5.microscopy.com
I have the Cressinton MTM10 thickness monitor. It has stopped working. Does anyone know anything about this monitor and what I can check other than the fuse which seems to be fine? OR does anyone know how to get in touch with Cressington. I have left phone messages and sent them email from their web site and have gotten no replies.
Thanks in advance for any suggestions.
Mei Lie -- Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
} We have been very happy with our Snap server otherwise known as an NAS } server. Basically just a bunch of hard drives and ethernet card. I have the } Quantum Snap with 240G of space mirrored. They are scalable and you can add } a bunch together to really serve some data. Users access their data in a } variety of ways, and it took about as long to set up as to unpack. It also } serves as the backup dump for the lab PC's.
I second the vote for the Snap server. Easy to set up and administer, plus our main unit network guru has ours set up to backup on another one in another building, while that one backs up on ours each night in case one building burns down...
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, I bought an EDAX model JSM-6100 detector for our lab through ebay at a good price. The information on the detector is as follows: 132-01, 10 mm2 active area, ser# 7455-57440ME, amp model#194-SUTW, PV9757/44ME.
It is the detector, connecting support arm, and dewar. It looks like it has an amplifier or electronic box on it as well. I am however missing the support electronics which go from the detector to the computer - wiring, software, computer card, and computer. If anyone has any experience with these detectors, or with installing them with homebuilt computers - let me know. It's a long term project I have to upgrade an Electronscan E3 ESEM to have analysis capability.
The detector looks like this one from EDAX: http://www.edax.com/products/Microanalysis/detectors/standard_EDS/10_liter.html
Thanks everyone.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
A couple of days ago I embedded some samples for microtomy, using Procure 812. Today when I went to remove the blocks from the mould I found that the mould material had adhered to the blocks and came away with the blocks, thus destroying two mould blocks. I have never experienced this problem in the past to such a degree.
The resin was measured and mixed properly and cured at 60 deg celcius. When embedding I half fill the mould and cure it in the oven at 60 deg celcius for an hour, then position the sample, fill the rest of the mould and back in the oven for final curing. This has worked in the past for me.
Does anybody have a better method of removing the cured blocks and not destroying the moulds, use a release agent etc etc??
Your help would be greatly appreciated.
Regards George
George Theodossiou Physicist / Electron Microscopist CSIRO Manufacturing & Infrastructure Technology Private Bag 33 Clayton South MDC Victoria, 3169 tel: +61 3 9545 2012 fax: +61 3 9544 1128
To the extent permitted by law, CSIRO does not represent, warrant and/or guarantee that the integrity of this communication has been maintained or that the communication is free of errors, virus, interception or interference.
The information contained in this e-mail may be confidential or privileged. Any unauthorised use or disclosure is prohibited. If you have received this e-mail in error, please delete it immediately and notify George Theodossiou on +61 3 9545 2012. Thank you.
George: My experience has been that the moulds age and that with repeated usage tend to start adhering to the cured blocks. At that point it's time to purrchase new moulds. I have had no success with release agents, etc. Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals Ridgefield, CT -- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } } A couple of days ago I embedded some samples for microtomy, using Procure } 812. Today when I went to remove the blocks from the mould I found that the } mould material had adhered to the blocks and came away with the blocks, thus } destroying two mould blocks. I have never experienced this problem in the } past to such a degree. } } The resin was measured and mixed properly and cured at 60 deg celcius. When } embedding I half fill the mould and cure it in the oven at 60 deg celcius } for an hour, then position the sample, fill the rest of the mould and back
} in the oven for final curing. This has worked in the past for me. } } Does anybody have a better method of removing the cured blocks and not } destroying the moulds, use a release agent etc etc?? } } Your help would be greatly appreciated. } } Regards } George } } George Theodossiou } Physicist / Electron Microscopist } CSIRO Manufacturing & Infrastructure Technology } Private Bag 33 Clayton South MDC } Victoria, 3169 } tel: +61 3 9545 2012 } fax: +61 3 9544 1128 } } Visit our Web site http://www.cmst.csiro.au } } Shipping address: CSIRO - Manufacturing & Infrastructure Technology, Gate 4 } Normanby Rd. Clayton, Victoria, } } PLEASE NOTE: } } To the extent permitted by law, CSIRO does not represent, warrant and/or } guarantee that the integrity of this communication has been maintained or } that the communication is free of errors, virus, interception or } interference. } } The information contained in this e-mail may be confidential or privileged.
} Any unauthorised use or disclosure is prohibited. If you have received this } e-mail in error, please delete it immediately and notify George Theodossiou } on +61 3 9545 2012. Thank you. } } }
Does anybody knows what is the acceptable variation in the extractor current on a LEO 1500 FE-SEM? Is it expected to observe a variation of the current for a Schottky FE-tip? Or might it be some external field? I am setting up a monitoring log now to see if this variation is periodic.
Our observed variation is about 3%.
Thanks in advance Sara
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} A histology question for the light microscopists out there: Are Haversian } systems present in the flat bones formed by intramembranous ossification or } are they only in long bones? Thanks, Tom
Tom:
Haversian systems are present in bone formed both ways. Haversian systems may be absent in very thin trabeculae of bone. In that case nourishment by diffusion through canaliculi is sufficient since the thickness of the trabecula is not greater than the diameter of a Haversian system.
Geoff
} Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
-- ********************************************** 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 mcauliff-at-umdnj.edu **********************************************
I have a geriatric JEOL 100C TEMSCAN that's older than dirt. It is under service contract with JEOL and the JEOL crew here in the Houston area have been phenomenally good about keeping it operating and operating well. They respond very quickly, at least within 24 hours. Now, that may be because we are in a large medical school in a large medical center and they have a lot of instruments in the area and are generally close by. It could also be because they just wander by my lab in a pavlovian reflex. I think they also understand that I run a diagnostic lab and patients don't stop getting sick just because Cartwright's scope is down. I am very happy with the service from JEOL.
Joiner Cartwright, Jr., Ph.D. Baylor College of Medicine Dept. of Pathology Houston, Texas U.S.A.
P.S. I have no financial ties to JEOL America, nor do I own stock in that company. However my car is parked on the 5th floor of Garage #6 today with the window down just enough for a plain brown envelope to slide through.
************************************************
At 06:03 PM 03/06/2003 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have any thoughts about the advantages/disadvantages of the use of various types of embedding molds (I'm referring to the substances from which they're made). If there's a preference for a certain type of material, then why?
We use Durcupan and EMbed/Araldite resins. I know that glycol methacrylate will not polymerize correctly in silicone molds, but does well in polyethylene. Other than that example, are there any problems in which the resin reacts with the mold material in any way?
Our "clear" silicone molds eventually turn opaque, with it happening faster around cavities that are used more often. Is this due to oven heat or the resin?
Thank you,
J.M. Lett Harold W. Siebens Hearing Research Center Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
My bosses are interested in sending me to a short course or workshop in digital imaging and image analysis. In particular they are looking for a workshop or short course which would be using Image-Pro Plus and Photoshop. Also geared toward biological EM applications. Anyone out there offering a short course or workshop along these lines?
Tom Bargar Core Electron Microscopy Research Facility 986395 Nebraska Medical Center Omaha, Ne 68198-6395
Hello George, When I was a graduate student at the University of Kansas, I was taught to spray the embedding molds with a teflon spray as one would do to a frying pan (I forget the brand name of the stuff). But for 15 years at the University of Iowa I have not done this and have had no problems removing blocks from "good" molds. When the molds begin to crack and the blocks become difficult to remove, I discard the mold. I have never experienced a sudden or overnight problem as you describe. Good luck.
Dean Abel Biological Sciences 141 BB University of Iowa Iowa City IA 52242-1324 USA
At 05:21 PM 3/7/2003 +1100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
AFter three Zr/O-W Schottky guns, my experience suggests that 3% variation is OK. I log Iext twice per month and perform gun brightness measurment every two months. I also record IPG and IPC pump currents.
These types (my type?) tend to rise in Iext after install and then stabilize at some certain current. At 92uA, mine varies by no more than +- 2uA. When the Iext starts to change more than this, that is an indicator that the gun is dying. Along with higher or lower Iext, gun chamber vacuum will deteriorate IPG will increase). When this happens, it's about two weeks to a totally dead gun. Typical life is about two years. My last one lasted exactly two years two months. some folks report half that lifetime. Keeping good vacuum and proper protocol for protecting the gun saves the cost of a new gun....or at least, puts it off longer.
gary g.
At 04:41 AM 3/7/2003, you wrote:
} Hello All } } Does anybody knows what is the acceptable variation in the extractor } current on a LEO 1500 FE-SEM? Is it expected to observe a variation of } the current for a Schottky FE-tip? Or might it be some external field? I } am setting up a monitoring log now to see if this variation is } periodic. } } Our observed variation is about 3%. } } Thanks in advance } Sara } } } -- } This message has been scanned for viruses and dangerous content by } MailScanner, and is believed to be clean. } } "The CSIR exercises no editorial control over E-mail messages and/or } attachments thereto/links referred to therein originating in the } organisation and the views in this message/attachments thereto are } therefore not necessarily those of the CSIR and/or its employees. } The sender of this e-mail is, moreover, in terms of the CSIR's Conditions } of Service, subject to compliance with the CSIR's internal E-mail and } Internet Policy."
I never used to have the problem with molds cracking or sticking to the resin but about 3 years ago it became a common occurence in my lab. I thought at first the supplier had changed the formulation but it was true from more than one supplier. Teflon sprays - available at your hardware store - help some but not much. I think using BDMA rather than DMP-30 in your epoxy resin mix makes it worse but that is not something I have carefully examined. Recently I switch to polyethylene "disposal" molds and they work pretty well - I re-use them but have only done this 3-4 times so I don't know what their lifespan will be.
} Hello George, } When I was a graduate student at the University of Kansas, I was } taught to spray the embedding molds with a teflon spray as one would do } to a frying pan (I forget the brand name of the stuff). But for 15 years } at the University of Iowa I have not done this and have had no problems } removing blocks from "good" molds. When the molds begin to crack and the } blocks become difficult to remove, I discard the mold. I have never } experienced a sudden or overnight problem as you describe. Good luck. } } Dean Abel } Biological Sciences 141 BB } University of Iowa } Iowa City IA 52242-1324 } USA } } } At 05:21 PM 3/7/2003 +1100, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
George, It sounds like your rubber mold has had one to many cures-the same happened to me with old rubber molds and Eponate 12.
Mike D ----- Original Message ----- } From: "George.Theodossiou-at-csiro.au"-at-sparc5.microscopy.com
Perfect timing. I was just about to post the following on this list. I think it exactly corresponds to your needs. We use Photoshop as a platform for the course because of its familiarity to a wide user base, but all of the plugins and techniques supplied are also fully compatible with Image Pro Plus. typically about 40% of the attendees are from the biological research community (other disciplines represented include materials science, medicine, forensic applications, geology, industrial imaging, etc.).
John Russ ===
The three-day intensive hands-on workshop on Image Processing and Measurement presented by John Russ (author of "The Image Processing Handbook" and "Practical Stereology") through the North Carolina State University Department of Continuing and Professional Education is now in its 21st year. The course dates for 2003 are May 21 - 23 in Raleigh, NC. This course has generated highly favorable reviews from the thousands of previous students. The primary focus is on images from various types of microscopy, with practical guidance in correcting imaging defects, enhancing the images for presentation and measurement, and performing stereological meaningful measurements on them. Textbooks and computer software are provided to attendees. Lab sessions with an opportunity to bring your own images makes this course immediately useful and highly productive.
For full information on the course, including outlines, faculty information, a downloadable brochure, and on-line registration, go to
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Class size is limited to maintain a high ratio of instructors to students, so make your reservation now. You may also contact Cindy Allen at NCSU Continuing Education, at 919-515-8171 ====
In a message dated 3/7/03 12:46:15 PM, tbargar-at-unmc.edu-at-sparc5.microscopy.com writes:
} My bosses are interested in sending me to a short course or workshop in } digital imaging and image analysis. In particular they are looking for } a } workshop or short course which would be using Image-Pro Plus and Photoshop. } Also geared toward biological EM applications. Anyone out there offering } a } short course or workshop along these lines?
George, You can use a TFE release agent to extend the life of your moulds, but the repeated heating, over time, causes the material to breakdown...what you experienced is pretty much inevitable. Moulds don't last forever. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Joiner, I don't know who you are, but I like your sense of humor. I'm thinking of trying the plain brown envelope trick with Leica and Philips. Let me know if it works. Mary Gail Engle
At 09:39 AM 3/7/03 -0600, Joiner Cartwright, Jr., Ph.D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
I would like to hear opinions or experiences using a Peltier cooled cryo-stage for SEM. Is this appropriate for high vacuum FEGSEM?
Greg
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295
Dear George, I always use a silicon release agent (a spray) when I embed samples. Sometimes they release, sometimes they don't or they take chunks of the mold with them. The molds have a life, but I make my own out of silicon rubber, so I just make some more. Good luck, 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 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {"George.Theodossiou-at-csiro.au"-at-sparc5.microscopy.c om} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 06, 2003 10:21 PM
We are looking for a technician with experience in clinical electron microscopy. Position Available immediately. Please contact me if you are interested know anyone who might be.
Lois Anderson Johns Hopkins University Dept. of Pathology Laboratory Manager Electron Microscopy/Immunofluorescence 600 N. Wolfe Street/Pathology 709 A Baltimore, MD 21287 (410) 955-2861/fax (410) 614-7110 landers-at-jhmi.edu
I was taught that going to Spot Size 1 in TEM gives more light at the expense of more beam damage to the specimen and lower resolution. In addition, I was told that one uses Spot Size 2 or 3 for high resolution work. Can one of the TEM experts out there expand on these simple statements. At what magnification does going to Spot Size 1 really begin to hurt resolution - if i am working at 20K, should it make a difference? I don't see much difference except in the amount of light. Comments and personal prejudices will be warmly accepted. If it makes any difference, I am working with conventional epon thin sections of biological material that was osmicated and UA and Pb counterstained; most photos are 20K or less but can sneak up to 40K or 50K. Thanks.
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Hi, I have two questions for image processing experts. A) how to change a gray scale image into a false color image. If the intensity of a gray scale image is in the range of 0 - 255, red, orange and blue color pixels in the corresponding color image should represent the pixels with intensity in the range of 0-100, 101 - 200, and 201-255 in the gray scale image, respectively. B) How to emerge three gray scale images into a color image. If the intensity of gray scale images is in the range of 0 - 255, red, orange and blue pixels in the color image should represent the pixels with intensity of 201-255 in the three gray scale images, respectively. Could you please tell me which software can solve the problems and how to do them? Your help is highly appreciated. Thanks, Jian-Guo
*********************************** Jian-Guo Zheng Research Assistant Professor Materials Science & Engineering Manager, NUANCE-EPIC http://www.nuance.northwestern.edu/epic/
Northwestern University 2220 Campus Drive, 1156 Cook Hall Evanston, IL 60208-3108, USA Phone: (847) 491-7807, Fax: (847) 491-7820 E-mail: j-zheng3-at-northwestern.edu http://www.nuance.northwestern.edu/epic/zheng/index.htm ***********************************
The problem of blocks sticking to the moulds might be the mould itself. They vary greatly in quality. We had to give up on one supplier because the moulds would start to deteriorate after only two or three usages. We are quite happy with the moulds supplied by Ted Pella (I have no financial interest in the company), but even these vary from batch to batch.
Ralph Common Michigan State University Division of Human Pathology
-----------------
Dear Colleagues,
A couple of days ago I embedded some samples for microtomy, using Procure 812. Today when I went to remove the blocks from the mould I found that the mould material had adhered to the blocks and came away with the blocks, thus destroying two mould blocks. I have never experienced this problem in the past to such a degree.
The resin was measured and mixed properly and cured at 60 deg celcius. When embedding I half fill the mould and cure it in the oven at 60 deg celcius for an hour, then position the sample, fill the rest of the mould and back in the oven for final curing. This has worked in the past for me.
Does anybody have a better method of removing the cured blocks and not destroying the moulds, use a release agent etc etc??
Your help would be greatly appreciated.
Regards George
George Theodossiou Physicist / Electron Microscopist CSIRO Manufacturing & Infrastructure Technology Private Bag 33 Clayton South MDC Victoria, 3169 tel: +61 3 9545 2012 fax: +61 3 9544 1128
Dear Colleagues, Most of grids are made of metals. Can anyone tell me if there are ceramic grids/slots/holes? Many thanks, Rong Yu
-- ************************************* Rong Yu Ph.D. Materials Science Division Lawrence Berkeley National Laboratory University of California Berkeley, CA94720, USA Phone: 1-510-486-6809 Fax: 1-510-486-7768 *************************************
The larger spot, the worse spatial coherence, and thus lower resolution for "High-Resolution" imaging. But such difference in resolution is hardly be detected at lower magnifications, e.g. 20k-50k.
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 was taught that going to Spot Size 1 in TEM gives more light at the } expense of more beam damage to the specimen and lower resolution. In } addition, I was told that one uses Spot Size 2 or 3 for high resolution } work. Can one of the TEM experts out there expand on these simple } statements. At what magnification does going to Spot Size 1 really begin to } hurt resolution - if i am working at 20K, should it make a difference? I } don't see much difference except in the amount of light. Comments and } personal prejudices will be warmly accepted. If it makes any difference, I } am working with conventional epon thin sections of biological material that } was osmicated and UA and Pb counterstained; most photos are 20K or less but } can sneak up to 40K or 50K. Thanks. } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
-- ************************************* Rong Yu Ph.D. Materials Science Division Lawrence Berkeley National Laboratory University of California Berkeley, CA94720, USA Phone: 1-510-486-6809 Fax: 1-510-486-7768 *************************************
In Photoshop, convert your image to indexed color:
Image, Mode, Indexed Color
In the Indexed Color dialogue box, choose:
Palette, Custom
You will see a grid of 256 squares. Click and drag over the first 85 squares and a color picker box will appear. Set those squares to the color you want by setting both the beginning and end square of the 85 squares to the same color. Do this three times for the three colors you want. Think about saving the indexed color profile so you don't have to do it again.
For your second question, I believe the Channels, Merge feature should be used. No time for details now but review the PhotoShop manual for this feature. We routinely use it to reconstruct 3 channel false color images from the confocal.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
} Hi, } I have two questions for image processing experts. A) how to change a } gray scale image into a false color image. If the intensity of a gray } scale image is in the range of 0 - 255, red, orange and blue color } pixels in the corresponding color image should represent the pixels with } intensity in the range of 0-100, 101 - 200, and 201-255 in the gray } scale image, respectively. B) How to emerge three gray scale images into } a color image. If the intensity of gray scale images is in the range of } 0 - 255, red, orange and blue pixels in the color image should represent } the pixels with intensity of 201-255 in the three gray scale images, } respectively. } Could you please tell me which software can solve the problems and how } to do them? Your help is highly appreciated. Thanks, } Jian-Guo } } } *********************************** } Jian-Guo Zheng } Research Assistant Professor } Materials Science & Engineering } Manager, NUANCE-EPIC } http://www.nuance.northwestern.edu/epic/ } } Northwestern University } 2220 Campus Drive, 1156 Cook Hall } Evanston, IL 60208-3108, USA } Phone: (847) 491-7807, Fax: (847) 491-7820 } E-mail: j-zheng3-at-northwestern.edu } http://www.nuance.northwestern.edu/epic/zheng/index.htm } ***********************************
I totally agree with Roger. You said, "etc. etc." I took this to mean you wanted a lot of help and info.
I use EPONATE 12 clone epoxy and the stuff releases fine when new PE molds are used. I have used flat silicone? embedding molds colored green and blue. This gets to be expensive when they give out. When they get old, the silicone rubber sticks as streaks on the blocks and wrecks the mold for any further use. They work fine for a time, just like the PE does. I have used my PE disposable molds, below, for over 8 years.
I use cheap Wheaton Bottle PE caps that can be purchased separately in different size diameters. (see below) I like a flat mold over the 'vertical' BEEM capsules that you can buy because of the types of samples I do and control of sample geometry. Anyway, after 5-10 uses, the epoxy sticks so bad that the polyethelene has to be trashed to get the round block out. I just throw the cheap mold away and use a new one. Hint: Just before the PE is going to stick, some crazing and slight sticking seems to take place. Also a thin or thick white area in the PE after removal signals the next embedding will be very hard to remove.
I believe the PE has some plasticizer on or near the surface that acts as a release agent. I could be wrong but there is something on the surface, in any case.
I use this large low profile mold because I do a lot of automotive coatings that contain TiO2 crystals and the larger molds are easier to pop the block out of. TiO2 trashes a diamond knife faster than most other things I run. This costs money and uses up diamond knife edge needlessly. So once I release a coil coat, 5 µm soda can ink or thicker 'mil' coating, I pin it down flat with straight pins that stick into the PE bottle caps and that hold the corners of the coating. Why use the pins? Sometimes the heat cure can cause the samples to curl during the curing and that also translates into excessive diamond knife edge useage and other problems in the TEM. I don't have to worry about a small sized sample being submitted. (The metal panels I get can be 10's of sq. inches in size.) If you only put a small amount of epoxy in at first, the raised number in the cap mold allows a thin film of epoxy to wick under the film even after pinning. After wetting, you can add the rest of the epoxy. This totally surrounds the embedded film or coating with epoxy. I saw the cured block with a X-ACTO RAZOR saw. It's faster but messy. Anyway, this flatness allows me to orient the straight and planar sample in the microtome so the smallest amount or profile is presented to the diamond knife edge used to thin section the coating for the TEM or for staining.
So why don't I use the small silicone molds? What I want to do is select the best flat area in the block that has the plane of the coating co-planar with the bottom of the mold. This also makes the coating co-planar with the jaws of the microtome vise mount on my RMC 6000 XL. This ensures that the diamond thin section cuts will be rectangular and not skewed. This is very critcal to the EDS examination of anti-reflective coatings on polymers.
These caps are listed in the regular Wheaton Catalog and can be ordered from Fisher Scientific using the Wheaton cap number.
I have never used Procure 812 but most clones of EPON 812 are the same. I have tested the ORIGINAL EPON 812 against Medcast (no longer available from Pella) and their Eponate 12 by having the NMR spectra run. Yes I still have a bottle of real EPON 812 from Shell ! NMR is not that sensitive as an analytical assay technique but the spectra match very close. The original stuff is thicker than Eponate 812 or the old Medcast. Obviously the molecular weight is now different in these newer clones. One other embedding kit I tested had minor levels of aromatics, FYI.
For cutting or trimming epoxy, use extra long Weck SS Razor Blades. They cost more but the pressure needed to be applied to cut the epoxy is 3-5 times less than a regular razor blade. This makes them safer to use from this stand point only. THESE blades CUT TISSUE like your FINGERS a lot easier also! They are extremely sharp and dangerous to use. They make a new regular razor seem dull. Take heed!!!! Never place your fingers or anything else below the edge of these blades.
I hope this helps give you some perspective and an overview of some hints on how others use Epon 812 clone epoxy and about silicone versus PE molds.
Paul Beauregard Senior Research Associate PPG Industries Monroeville Chemicals Technical Center 440 College Park Drive Monroeville, PA 15146 724-325-5131
} } George: } My experience has been that the moulds age and that with repeated usage tend to } start adhering to the cured blocks. At that point it's time to purrchase new } moulds. I have had no success with release agents, etc. } Roger Moretz, Ph.D. } Dept of Toxicology } BI Pharmaceuticals } Ridgefield, CT } -- } } } Dear Colleagues, } } } } A couple of days ago I embedded some samples for microtomy, using Procure } } 812. Today when I went to remove the blocks from the mould I found that the } } mould material had adhered to the blocks and came away with the blocks, thus } } destroying two mould blocks. I have never experienced this problem in the } } past to such a degree. } } } } The resin was measured and mixed properly and cured at 60 deg celcius. When } } embedding I half fill the mould and cure it in the oven at 60 deg celcius } } for an hour, then position the sample, fill the rest of the mould and back } } } in the oven for final curing. This has worked in the past for me. } } } } Does anybody have a better method of removing the cured blocks and not } } destroying the moulds, use a release agent etc etc?? } } } } Your help would be greatly appreciated. } } } } Regards } } George } } } } George Theodossiou } } Physicist / Electron Microscopist } } CSIRO Manufacturing & Infrastructure Technology } } Private Bag 33 Clayton South MDC } } Victoria, 3169 } } tel: +61 3 9545 2012 } } fax: +61 3 9544 1128
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mwb142-at-psu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, March 7, 2003 at 12:42:42 ---------------------------------------------------------------------------
Email: mwb142-at-psu.edu Name: Matt Bell
Organization: Penn State
Education: Graduate College
Location: State College, PA
Question: I know that there is a Sparrow approximation for resolution of an optical wave. Does that apply for acoustic waves? And if so, what is the formula for the Sparrow approximation for lateral and vertical resolution?
What you are suggesting is not the only way. Of course you can create a false color image, but you can just as easily create a full color image by assigning each of the three colors to one color channel of the full color (24-bit) image. If you really do need a false color image, you can simply convert this image. The results are likely to be better as any reasonable software will optimize the color palette.
As for software, look for software that supports Fluorescence imaging (such as, but not limited to, our analySIS software). Your question is a standard problem in fluorescence, as often a b/w camera is used (because of higher sensitivity) to acquire several fluorescence channels and merge them into a color image. Often it is more than 3 colors, which makes it more complicated, but it is still a standard technique (multiple Fluorescence) for many software packages.
Please contact me offline if you want more information.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Jian-Guo Zheng [mailto:j-zheng3-at-northwestern.edu] Sent: Friday, March 07, 2003 4:56 PM To: Microscopy-at-sparc5.microscopy.com Cc: 'Jian-Guo Zheng'
Hi,
We have 3 SEM's and 2 TEM's in our facility with varying costs and amounts of use associated with each. Using the Cost/Hours of Use=Rate leaves us with scopes having unappealingly high rates simply because they aren't used much or have higher annual costs. Your heads are nodding in agreement now, right?
What can't I use an "average" rate for SEM's and TEM's that (somewhat) smooths out those irregular use and costs patterns? Of course I can, right? Well, in truth it would help me to be able to tell Research Accounting that others are doing it too :} ).
Is anyone else doing this or something like it?
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Thank you for all the replies and useful hints, for release agents and different types of moulds.
I understand that moulds have a finite life but what really annoyed me about this instance was that the moulds were white silicone rubber, had only been used 3-4 times previously and had very few cracks. I thought they had a bit of life left in them. Oh well, it just makes work more fun.
Regards George
George Theodossiou Physicist / Electron Microscopist CSIRO Manufacturing & Infrastructure Technology Private Bag 33 Clayton South MDC Victoria, 3169 tel: +61 3 9545 2012 fax: +61 3 9544 1128
To the extent permitted by law, CSIRO does not represent, warrant and/or guarantee that the integrity of this communication has been maintained or that the communication is free of errors, virus, interception or interference.
The information contained in this e-mail may be confidential or privileged. Any unauthorised use or disclosure is prohibited. If you have received this e-mail in error, please delete it immediately and notify George Theodossiou on +61 3 9545 2012. Thank you.
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what is the timescale of these variations? In case seconds, then there is something wrong with your filament ... this may be fixed with an bakeout and a "reconditioning": trying to find a new extractor voltage and filament current is usually the last measure before exchanging the filament I think the DENKA's behave a bit different compared to the one's from FEI ...
There is also some further reading which may be helpful for understanding: F. Zhou:"Handbook of charged particles optics", pp.77-102 "A Review of the ZrO/W Schottky Cathode", CRC Press
But the main point to check the status of your GEMINI is the probe current: With a 30µm aperture -at- 1kV our's have been at 90..130pA (...as long as the aperture is o.k.). The DENKA we are using right now has about 5500h and has been at more than 180µA extractor current (...EHT on) more or less form the beginning!!! On the second to minute timescale the variations are definitely below 0,5%(!!!) ... o.k.: we are doing only LV-morphology work (...no EDX/WDX) ... and we do not change the extractor voltage from the set working point of 6KV for more than +/- 400V for longer than 4h ... IGP vac always better than 2x10^-9´mbar
If the variation (... always talking about the time after having reached the saturation, that means 4..6h after switching on the gun!!!) is periodically on a minute to hour scale, there is something wrong with the ZrO droplet which should wet the tip of the W single crystal ...
Hope this helps a bit?
Gunnar
Date sent: Fri, 07 Mar 2003 10:02:19 -0800 To: "Sara Prins" {SPrins-at-csir.co.za} } From: Gary Gaugler {gary-at-gaugler.com}
Owen, I ran into the same problem when we set up our rate structure last year. Accounting wanted each scope treated separately based on it's use and service contract costs. This amounted to vastly different rates for our three scopes. We intend to go back and argue for the need to treat all as one unit and combine maintenance costs plus use hours to come up with a reasonable hourly rate for all. Our internal people are fortunate in that the rate will be heavily subsidized for most of them, make it very affordable. It will affect external users and some unsubsidized university users the most but should end up being more fair overall than the way it is now. Hopefully this will be accepted by the university accounting office.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 3/9/03 4:35 PM, "Owen P. Mills" {opmills-at-mtu.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, } } We have 3 SEM's and 2 TEM's in our facility with varying costs and amounts } of use associated with each. Using the Cost/Hours of Use=Rate leaves us } with scopes having unappealingly high rates simply because they aren't } used much or have higher annual costs. Your heads are nodding in } agreement now, right? } } What can't I use an "average" rate for SEM's and TEM's that (somewhat) } smooths out those irregular use and costs patterns? Of course I can, } right? Well, in truth it would help me to be able to tell Research } Accounting that others are doing it too :} ). } } Is anyone else doing this or something like it? } } Owen } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills } } } }
I can not answer your specific question about whether others are using average cost. However, as a recent graduate from business school, and one who three times attended summer hockey camp at your university (Go Huskies!) I offer some input that I hope you find helpful.
Accounting costs are an approximation of opportunity cost, and to the extent that they reflect opportunity cost they serve as a fair measure for transfer pricing (of services purchased within the organization) or a useful measure for external pricing (of services sold outside the organization).
Opportunity cost indicates the value that you might extract from your tool given the options that you have. Options may include performing research, charging an external customer, and selling the tool. In an extreme, if you choose one option, others are foregone. If you strictly perform research, for example, your opportunity cost is the difference between the present value of performing that research and the greater of charging external customers or selling the tool. You might have reason to argue that you are doing the right thing and to do so by stating alternatives, and noting that those alternatives provide less value than your desired use.
Opportunity cost may be the marginal cost of operating a tool. Marginal costs would be those costs that occur only when you turn the tool on, excluding those costs which are otherwise committed to, such as purchase price of the tool and cost to maintain the building. Considering another extreme, it may be that your highest and best use of this tool and the facility would be to rent it out at a fee merely greater than the cost of electricity to run the tool.
In contrast to opportunity costs of today, your organization's accounting costs are probably what is required to recover dollars already spent to purchase the unit and to operate the facility that it houses it. Indeed, this would reflect the opportunity cost at the time the tool was purchased. However, such accounting may reflect little of today's economic situation and in the worst case it leads to failure to extract the most possible value from the tool.
While you may find standard practices to support use of "average pricing", I hope that you might also consider opportunity cost as a method for both decision making as well as persuasion. Finally, I recommend an easy reading novel that provides an understanding of standard accounting practices and suggests a framework for extracting maximum value from an organization's assets, "The Goal", by Elijahu M. Goldratt.
Sincerely,
John Moore Montara Industries New Hill, North Carolina 919-434-8457
"Assembling a team to provide 3rd party support for the Applied Materials SEMVison."
----- Original Message ----- } From: "Owen P. Mills" {opmills-at-mtu.edu} To: "Microscopy" {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, March 09, 2003 4:35 PM
Hi,
We have 3 SEM's and 2 TEM's in our facility with varying costs and amounts of use associated with each. Using the Cost/Hours of Use=Rate leaves us with scopes having unappealingly high rates simply because they aren't used much or have higher annual costs. Your heads are nodding in agreement now, right?
What can't I use an "average" rate for SEM's and TEM's that (somewhat) smooths out those irregular use and costs patterns? Of course I can, right? Well, in truth it would help me to be able to tell Research Accounting that others are doing it too :} ).
Is anyone else doing this or something like it?
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
The Connecticut Microscopy Society is pleased to present Dr. Pietro De Camilli on Thursday, March 27 at the Peabody Museum at Yale University. Dr. De Camilli will describe his work on synaptic transmission. A reception and dinner precede the lecture. Information is on our website, http://www.connms.org We hope to see you there!
Elizabeth Cowles, President of the Connecticut Microscopy Society
Elizabeth A. Cowles, Ph.D. Associate Professor, Department of Biology 220 Goddard Hall Eastern Connecticut State University 83 Windham St. Willimantic, CT 06226 voice: 860-465-4385 fax: 860-465-5213 email: cowlese-at-easternct.edu home page: http://www.easternct.edu/personal/faculty/cowlese/index.html http://biology.easternct.edu
To amplify what Geoff has said slightly, I would only add that "FIRST" dermal bone is often called "woven bone" (non-lamellar in its initial structure) and may even become known as "spicular or trabecular" bone thereafter before it becomes remodeled as lamellar bone. The reason I mention this is that if one is looking at embryonic/fetal bone, then one may not see haversian systems or periosteal or endosteal lamellar bone at the time of sampling. Furthermore, there are even some small fish whose bone is actually acellular in the adult (as I recall dimly from coursework taken years ago). Some of the smaller sesamoid bones may also lack haversian and/or lamellar bone, although without looking I would bet that either or both of my patellas do. The haversian system is considered the fundamental unit of bone remodeling and is often referred to as an 'osteon'.
It would be relatively easy to distinguish between the lamellae of periosteal and endosteal bone from those in haversian bone. Indeed, one can often observe between periosteum and endosteum the arcs of remnant older haversian systems between the newer, uninterrupted osteons.
It would also be worthwhile mentioning that the axes of collagen fibrils are parallel within a single lamella and opposing in adjacent lamellae [mindful of micelles of cellulose in adjacent layers of wood growth] and not described as being paraxial with the long axis of a long bone. Thus, perfect transections of long bone viewed with the TEM will never show transverse sections of collagen micells without some tilt.
The most recent really good basic histology book is the 12th edition of Bloom and Fawcett, published in 1994 (ISBN: 0-412-04691-1). There should be some of this latest in a classic series of higher-end biology books available on the used book market. I got mine as new just last year.
Hope this helps too,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging
Mail to: Geology, CASI West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383
For help and information only, The CASI houses: An FEI Quanta 400 and Technai 12T, Oxford INCA Energy 400, Tousimis AutoSamdri 815 and Olympus FV-300.
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Friday, March 07, 2003 10:00 AM To: Tom Phillips Cc: Microscopy-at-sparc5.microscopy.com
Tom Phillips wrote:
} A histology question for the light microscopists out there: Are Haversian } systems present in the flat bones formed by intramembranous ossification or } are they only in long bones? Thanks, Tom
Tom:
Haversian systems are present in bone formed both ways. Haversian systems may be absent in very thin trabeculae of bone. In that case nourishment by diffusion through canaliculi is sufficient since the thickness of the trabecula is not greater than the diameter of a Haversian system.
Geoff
} Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
-- ********************************************** 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 mcauliff-at-umdnj.edu **********************************************
On Friday, March 7, 2003, at 04:55 PM, Rong Yu wrote:
} The larger spot, the worse spatial coherence, and thus lower } resolution for } "High-Resolution" imaging. } But such difference in resolution is hardly be detected at lower } magnifications, e.g. 20k-50k. } } Tom Phillips wrote: } } } I was taught that going to Spot Size 1 in TEM gives more light at the } } expense of more beam damage to the specimen and lower resolution. In } } addition, I was told that one uses Spot Size 2 or 3 for high } } resolution } } work. Can one of the TEM experts out there expand on these simple } } statements. At what magnification does going to Spot Size 1 really } } begin to } } hurt resolution - if i am working at 20K, should it make a } } difference? I } } don't see much difference except in the amount of light. Comments and } } personal prejudices will be warmly accepted. If it makes any } } difference, I } } am working with conventional epon thin sections of biological } } material that } } was osmicated and UA and Pb counterstained; most photos are 20K or } } less but } } can sneak up to 40K or 50K. Thanks. } } Dear Tom and Rong, Another consideration for very high mags is that the closer the beam is to crossover, the poorer the resolution, so one may want to use a smaller spot size number (actually, bigger spot) further from crossover rather than a bigger spot size number at crossover. Also, your specimen may be better behaved vis-à-vis charging/heating induced beam movement if a larger area is illuminated. As you said, Rong, all this is irrelevant at 20k-50k. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Just some last minute comments from a long-time mold manufacturer:
Standard blue silicone rubber molds - These are quite pliable. We've found the life span is mainly dependent upon the number of cures and components of the epoxy mixture. Over-curing at high temps can also limit lifespan.
Clear silicone rubber molds - These require a different silicone formulation. They do tend to crack when bent. Obviously you have to balance this with the bottom illumination advantage.
We've looked at other rubber molds manufactured in the U.S. and most use pretty good material. Our labs do use our TFE spray which helps in certain situations and it is now environmentally-friendly.
We also do a lot of custom silicone molds and some are quite large. These may require modification of curing procedures.
John Arnott Disclaimer: Ladd sells EM supplies and accessories
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
Does anyone know of an opaque white resin that can be used for tissue infiltration? We want to progressively section and photograph the block face
Thanks Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University, Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 http://www.anu.edu.au/EMU
Is anyone using an automatic negative/film developer for TEM negatives that they are happy with? Our EM Lab is looking for an auto developer that would develope Technical Pan Film TP-120 (a roll of approx 8 negatives that is taken with a Zeiss TEM 109). Manufacturer and price will be most helpful. Any imput will be appreciated. Many thanks to all who respond or take precious time to read. Teresa
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rajmeister-at-msn.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, March 10, 2003 at 22:49:28 ---------------------------------------------------------------------------
Email: rajmeister-at-msn.com Name: Raj
Organization: Mary Washington College
Education: Undergraduate College
Location: F'brg, VA, USA
Question: Hello, My question is, what do you think about the Observer IV microscope overall for its price($265)? And also what is the procedure for preparing an oil immersion slide for use at 1000x? Thanks
We are the latest folks to get caught by the "New Formulation" TEM films. We had a couple of batches come out of our nitrogen-burst processor looking like they'd been tossed into a vat of developer and left untouched for a couple minutes. The negs were way light and mottled with streaks and patches indicative of poor agitation. Unfortunately, I didn't even realize we had any of this film in our inventory, since we hadn't bought any since before the controversy hit the listserver, so our poor student lab assistant got the blame.
When we finally hit on the cause of this, we were able to remedy it by increasing our agitation cycle to 2 seconds every 10 seconds, instead of 2 seconds every 30 seconds (which worked beautifully with the old film). We use a 4-minute development in d-19 at 68-70 degrees F, and we may move to 5 minutes. The agitation marks have been eliminated and density is back into the good range, although still a bit lighter than before. We had also started to get a "thumbprint" mark on the bottom of the negatives which puzzled us greatly until our faculty coordinator pointed out that it coincided exactly with the round plastic rod that the negatives rest on at the bottom of the rack. It was causing increased local agitation in a half-oval pattern as the nitrogen bubbles swirled around it. Lifting the negatives up slightly seems to have cured this. For some reason we had never noticed it before.
In case you were curious, the student lab assistant has been duly apologized to and was very gracious in his victory.
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Consult a professional photographer who is familiar with the type of job you are doing. Proper lighting coupled with a specific film and processing may allow tissue in a paraffin or epoxy block to stand out from the background. You might also try mixing something into your embedding material to make it opaque.
Geoff
Sally Stowe 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 an opaque white resin that can be used for tissue } infiltration? We want to progressively section and photograph the block } face } } Thanks } Sally } } Dr Sally Stowe } Facility Coordinator, ANU Electron Microscopy Unit } Research School of Biological Sciences } Australian National University, Canberra ACT0200 } AUSTRALIA } stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 } http://www.anu.edu.au/EMU
-- ********************************************** 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 mcauliff-at-umdnj.edu **********************************************
I am looking for a statement of acceptable water quality for TEM usage. This may be familiar to those of us who are *blessed* to be inspected by CAP. All I know about water quality is when it's NOT! Does anyone have a statement that they use that they would be willing to share?!
Thanks,
Karen Bovard EM Lab Creighton University Medical Center Omaha, Nebraska
The Oklahoma Microscopy Society and the Central States Microscopy and Microanalysis Society will be holding a joint meeting in Tulsa, Oklahoma on April 10-11, 2003. The theme of tis meeting is "The Art of the Science Image."
Thusday, April 10 Session I: Image Creation 9:00-9:50 Opening Address: "Images and Human Understanding", John Russ, NCSU
10:00-10:45 "A Review of X-Ray Mapping", John Friel, PGT
10:45-11:00 BREAK
11:00-12:00 "Spectrum Imaging: The Next Step in Microanalysis", Paul Kotula, Sandia Labs
12:00-1:00 LUNCH
1:00-1:30 "Winning the Depth of Field Battle in Light Microscopy", Jim DeMian, 3M Co.
1:30-2:30 "Specialized Methods in Light Microscopy", Robert Weaver, McCrone Inst.
Evenong Reception - IMAX Theatre 6:00-7:00 Buffet Dinner 7:00-8:30 "Four Million House Guests" a.k.a. "The Hidden Dimension", 3D movie and Keynote Addrerss: "SEM Stop-Frame, Color, 3D Animation for Motion Pictures", David Scharf, Scarf Photography
Friday April 11 Session II: Image Processing and Analysis 8:30-12:30 "Guide to Image Analysis Workshop", John Russ, NCSU
12:30-1:30 LUNCH
Session III: Image Presentation 1:30-2:15 "Basic Principles of Image Composition for Scientists", TU Art Dept.
2:15-2:30 BREAK
2:30-3:30 "Exploring the Invisible: the Cultural Impact of Microscopy", Lynn Gamwell, SUNY
Special Features Corporate Exhibitors Demonstrations of image processing software Region-wide "Art of the Science Image" competition Art Gallery displays "Images from Science", Rochester Inst. of Tech and from David Scharf
Hotel Accommodations Renaissance Tulsa, the city's newest hotel, located across from the IMAX theatre, special meeting rate of $84 per night for single or double; complimentary pool and fitness facilities; complimentary shuttle from airport and to nearby shopping mall.
Registration In advance ( {3/31), $40 for members, $50 for non-members, $20 for students On-site (} 3/31), $50 for members, $60 for non-members, $25 for students Thursday night reception: $20 note: non-member registration includes a 2003 membership to either OMS or CSMMS
For more information or to receive a registration packet, contact Lou Ross at rosslm-at-missouri.edu or at (573) 882-4777.
-- Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211-5120 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.biotech.missouri.edu/emc
Leslie, We have been in the very fortunate position to have extensive university support to cover costs associated with this microscopy facility. The administration, primarily the School of Agriculture, has provided sufficient funds to cover service contracts on 3 EMs and additional funds for other operating costs. In addition salaries of two full-time professional level positions are covered by schools of Agriculture, Science, and Veterinary Medicine since this core facility caters primarily to researchers within those schools. This means that for many years we had no charges for use of the facility other than to cover consumables such as film. The facility encourages multi-user access but also does provide full service (there were always charges for service).
Under current financial stress, we have instituted a rate system to try to cover future increases in costs as well as to try to reclaim some of the current operating costs. Salaries are not figured into the recharge except for the service option. Administrators here understand that EM facilities do not make money...they are fortunate to recover most of their costs but even this is difficult if you want the facility used by the greatest possible number of researchers.
Now, that is the brief history behind the facility funding. It is true that you must charge identical amounts to all users of a facility if any are paying for costs using grant funds. You cannot preferentially exempt some people and supplement others. We use the subsidies to keep our rates low for all users. By doing this there is also the commitment by the administrators to continue the subsidies and not expect the facility to cover all associated costs. Everyone within the university who are doing sponsored research or unfunded research pay the same amount.
However, in some cases researchers do proprietary work on contract with private companies. In this case, they are not able to share the results with the research community so are not entitled to subsidized rates. At the present time they pay the full amount based on actual costs divided by use. Since the use is by university staff, no indirect charges are applicable. This also applies to companies who have direct ties to the university such as those who are industrial affiliates of departments or programs (in which case they are paying a fee for access to university staff and facilities).
Those companies who do not have formal ties to the university pay the unsubsidized rates plus indirect costs to the university to cover infrastructure costs. They also would pay consultant costs for assistance by university staff.
We have found so far that keeping rates low through the subsidies increases the number of users. Since the equipment does no good if unused, this is very desirable. In return, as the number of users increases, so does the revenue. The revenue then goes to "pay back" funds used for the subsidies. I would guess that we generate more overall revenue with the lower rates and more users benefiting from the facility than if we charged double the cost and user numbers fell accordingly. Its like anything else...if users perceive a bargain they will come...and often spend more than they originally intended because they feel they are getting value for their $.
Now, some equipment has been purchased by asking university faculty to contribute required funds. This is the case for some of our light microscopes and computer equipment. In this case, when these researchers agree to help fund facility equipment, they understand that access is on a first-come basis. In return the equipment is maintained and students are trained in the use by facility staff. The researchers individually put in much less money than if they were purchasing the entire instrument so we usually end up getting higher end equipment. There are no fees associated with using this jointly funded equipment.
Major equipment is funded almost entirely through instrumentation grants with required matches from the university. Researchers who assist in writing these grants understand that they are doing this partially as a service to the university community. They do not get preferential treatment if the instrument resides in the common core facility but usually end up using the equipment most since they had the research justification for it in the first place.
Devising a fair rate system is a complicated business and we have found that we need to clarify and revise as the system reveals the need.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 3/10/03 11:53 AM, "Leslie Eibest" {leibest-at-duke.edu} wrote:
} Hi Debby, } } I'm very interested in the method used there to subsidize some } users. Currently, we have a couple of departments that subsidize my salary } and the service contract, and those department members have free use of the } equipment and technical assistance. Outsiders were charged reasonable } rates. Everyone loved the system, and it provided a lot of stability for } the SEM lab. However, Sponsored Programs has come down on our necks, saying } that this unfairly subsidizes some grant holders and not others. All } federal grant holders should be charged the lowest rate. We will have to } raise more funds from user fees, so now we'll have to charge } everyone. Researchers and students without funding won't be able to use } the equipment. Adding to the fun, they won't let me keep reserves for } equipment repairs and upgrades. It's ironic, since NSF was pleased that } our lab was accessible to so many users. } } Have you had to deal with these issues yet? If so, how do you get around } the problems? } } Leslie } } } } } We intend to go back and argue for the need to treat all as one unit and } } combine maintenance costs plus use hours to come up with a reasonable hourly } } rate for all. Our internal people are fortunate in that the rate will be } } heavily subsidized for most of them, make it very affordable. It will } } affect external users and some unsubsidized university users the most but } } should end up being more fair overall than the way it is now. Hopefully } } this will be accepted by the university accounting office. } } } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-052 Whistler Building } } 170 S. University Street } } West Lafayette, IN 47907 } } } } } } On 3/9/03 4:35 PM, "Owen P. Mills" {opmills-at-mtu.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, } } } } } } We have 3 SEM's and 2 TEM's in our facility with varying costs and amounts } } } of use associated with each. Using the Cost/Hours of Use=Rate leaves us } } } with scopes having unappealingly high rates simply because they aren't } } } used much or have higher annual costs. Your heads are nodding in } } } agreement now, right? } } } } } } What can't I use an "average" rate for SEM's and TEM's that (somewhat) } } } smooths out those irregular use and costs patterns? Of course I can, } } } right? Well, in truth it would help me to be able to tell Research } } } Accounting that others are doing it too :} ). } } } } } } Is anyone else doing this or something like it? } } } } } } Owen } } } } } } Owen P. Mills } } } Electron Optics Engineer } } } Materials Science & Engineering } } } Michigan Technological University } } } Rm 512 M&M Bldg. } } } Houghton, MI 49931 } } } PH 906-369-1875 } } } FAX 906-487-2934 } } } mailto:opmills-at-mtu.edu } } } http://www.mm.mtu.edu/~opmills } } } } } } } } } } } } } }
Oops, sorry. My posting was about the 4469 film. Thanks for the reminder, Mike.
Randy
-----Original Message----- } From: Mike Coviello [mailto:coviello-at-mae.uta.edu] Sent: Tuesday, March 11, 2003 12:52 PM To: Tindall, Randy D.
To all, The Core Facility Management session at the annual Microscopy and Microanalysis meeting is a forum to discuss topics relating to the management issues associated with core facilities in both the academic and private sector. The format has been to have a presenter to introduce the topic and then having an interactive discussion session. Many of the proceedings have been taped, transcribed and published in Microscopy Today so that those who could not attend the meetings still had the benefit of the discussions. Some of the topics discussed in past sessions include:
Managing Users Justification of costs/cost recovery Equipment maintenance issues Training Users Calibration of Ems Scientific Ethics and responsibilities of facility managers.
I must finalize the topic(s) for M&M 2003. As we want to make this session timely, I would like input from readers on topics of current interest. This year we will receive no support from M&M 2003 but they have promised us a room and audio-visual equipment.
Please respond immediately with topic suggestions.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
First, let me assure you I know how to spell cryostat! The listserver spam filter blocks messages with CRY (as in Cry for Help) in them and Nestor is working on it. In the meantime I thought I would try this work around. I have several questions for the cryostat gurus out there.
First, how do you decide whether to infiltrate with sucrose? I will be fixing my tissue with 2% paraformaldehyde and some protocols call for direct freezing and others for infiltration with 30% sucrose in PBS first. I know the sucrose will cryo-protect and suspect it will also improve the plasticity but is there a disadvantage? A minor side question is whether the 30% sucrose can be simply made up in 1x PBS or do you need to account for the change in volume that the sucrose will probably cause. Presumably osmolarity is no longer a big issue if you are using 30% sucrose.
Second, what is the feeling on knives - high profile vs low profile? Is it worth paying extra for the heavy duty ones to minimize chatter or is that only needed for hard tissues. I will be cutting lymph nodes.
Thanks for any tips. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
} First, how do you decide whether to infiltrate with sucrose?
If you are going to freeze the tissue cryoprotection is a good idea. If your samples are small and you freeze quickly, you may be able to get away w/o cryoprotection.
} I will be } fixing my tissue with 2% paraformaldehyde and some protocols call for } direct freezing and others for infiltration with 30% sucrose in PBS } first. I know the sucrose will cryo-protect and suspect it will also } improve the plasticity but is there a disadvantage?
Not that I know of.
} A minor side question } is whether the 30% sucrose can be simply made up in 1x PBS or do you need } to account for the change in volume that the sucrose will probably } cause.
I don't. I have used sucrose concentrations from 20% to 30% with no apparent differences so if you are off a bit, no problem. What is important is the speed of freezing, the faster the better.
} Presumably osmolarity is no longer a big issue if you are using 30% } sucrose.
Nope. At least not in my experience.
} Second, what is the feeling on knives - high profile vs low profile? Is it } worth paying extra for the heavy duty ones to minimize chatter or is that } only needed for hard tissues. I will be cutting lymph nodes.
I can't help you there, I usually cut rodent brains with plain old microtome knives.
} Thanks for any tips. Tom } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
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 mcauliff-at-umdnj.edu **********************************************
This is perhaps a bit unusual of a topic but it is centered on EDS viability for foiling double blind testing.
Question: Can EDS be used to analyze a double blind tablet to ascertain whether it is placebo or a study medication?
It would seem that the inability to see light element H might not be a limiting factor. Are there signatures of placebos that would strongly differentiate them from study meds? How might one guard against educated analysis of study meds? If the basic elements are C, H and O, are there distinct signatures or quantities that one could use to distinguish between placebo and study meds?
Randy, I'm glad you apologized to your student...its very frustrating to be lamed for someting that is totally out of your control! Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
HI Tom, The preference in our facility is to cryoprotect with sucrose whenever possible. We even freeze our tissues in a 1:1 mixture of sucrose (20% in PBS) and OCT. It makes a softer block, and you may have to drop your chamber temp to -20, but it cuts very smoothly and yields very nice structure. The only time we don't use sucrose is when the primary Ab's to be used won't tolerate any fixation, then we snap freeze and pray. Our protocol is a modification of that published by Barthel & Raymond in J Histochem Cytochem 3899) 1383-1388 1990. They were looking at eyes. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I am not sure what is the film thickness you need to cross-section, if it's over 10 microns it might be quite difficult. However, we have successfully cross-sectioned individual multi-walled carbon nanotubes and bundles of those (on SiO2/Si substrates) using combination of FIB 'lift-out' and broad beam ion milling in Gatan Ion Mill. At the first stage, you use the FIB 'lift-out' technique to make a sample with the thickness 0.8-1.0 microns. The sample is then mounted with the glue on the edge (on a bar) of the Cu half-grid and subjected to ion milling (4 kV, followed by 2 kV) in Gatan Ion Mill. Typical milling time in Gatan: 4-6 minutes. Keep checking the sample thickness in TEM and continue milling as necessary. The sample quality is good enough for high resolution TEM and EELS.
Let me know if you are interested in more detail, I can send you our recent paper.
Regards, Katharine ******************************* Katharine Dovidenko Assistant Professor School of NanoSciences and NanoEngineering & UAlbany Institute for Materials University at Albany-SUNY 251 Fuller Rd., Albany, NY 12203 Phone: (518) 437-8781
-----Original Message----- } From: yimin yao [mailto:yimin-at-fy.chalmers.se] Sent: Wednesday, March 12, 2003 3:49 AM To: Microscopy-at-sparc5.microscopy.com
Dear colleagues,
Does anyone have experience to use FIB to make cross-section TEM samples for a carbon nanotube film on substrate?
Best regards,
Yiming
------------------------------------ Dr. Yiming Yao
Microscopy and Microanalysis Department of Experimental Physics Chalmers University of Technology SE-41296, Göteborg Sweden
UVM Practical Course on Three-dimensional Cryo Electron Microscopy of Single Particles
August 11-17, 2003 Burlington, Vermont
The course will teach the principles of three-dimensional reconstruction of single particles from electron micrographs.
It includes demonstrations of the experimental aspects, teaching of the basic theoretical principles and six hours per day of hands on experience in processing data sets from cryo-electron microscopy images.
Participants will work in groups of two and six instructors will be available to guide everyone step by step through the complete reconstruction process. Four participants will have the opportunity to carry out practical microscopy work the two days following the end of the course.
For further information and application visit our web site: http://physioweb.med.uvm.edu/Cryo_Practical/
Organizers and Teachers:
Michael Radermacher Teresa Ruiz Montserrat Barcena Jean Francois Menetret Montserrat Samso T.B.A.
__________________________________________________ Michael Radermacher, Assoc. Prof. University of Vermont College of Medicine Department of Molecular Physiology and Biophysics HSRF Building Burlington, VT 05405
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Study medications would most likely contain detectable amounts of nitrogen. There might also be Na or K.
-Ken Gaugler
Gary Gaugler 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 perhaps a bit unusual of a topic } but it is centered on EDS viability for } foiling double blind testing. } } Question: Can EDS be used to analyze } a double blind tablet to ascertain whether } it is placebo or a study medication? } } It would seem that the inability to see } light element H might not be a limiting } factor. Are there signatures of placebos } that would strongly differentiate them } from study meds? How might one guard } against educated analysis of study meds? } If the basic elements are C, H and O, are } there distinct signatures or quantities } that one could use to distinguish between } placebo and study meds? } } gary g. } } }
On Tuesday, March 11, 2003, at 06:16 PM, Gary Gaugler wrote:
} This is perhaps a bit unusual of a topic } but it is centered on EDS viability for } foiling double blind testing. } } Question: Can EDS be used to analyze } a double blind tablet to ascertain whether } it is placebo or a study medication? } } It would seem that the inability to see } light element H might not be a limiting } factor. Are there signatures of placebos } that would strongly differentiate them } from study meds? How might one guard } against educated analysis of study meds? } If the basic elements are C, H and O, are } there distinct signatures or quantities } that one could use to distinguish between } placebo and study meds? } Dear Gary, Most medications consist of small amounts of the active ingredient mixed with a much larger amount of matrix (which provides sufficient volume to handle a tablet, gets the active ingredient into the stomach--and sometimes further along the digestive tract--dissolves at a rate appropriate to deliver the drug as desired, etc.). The placebo in a properly designed trial consists of matrix identical to that for the drug, so EDS analysis would almost certainly give the same result for both. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
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In your message you only mention elements you believed would be present in the placebo tablet. However as Ken Gaugler alluded to, if you had information regarding the chemical composition of the active ingredient you might be able to ascertain if it were absent in a placebo tablet. For example, if the active contained sulfur EDS would (likely) be able to detect the sulfur in a tablet assuming the excipients/binders (see response from Bill Tivol) did not contain sulfur. However, unless you can verify this on a tablet containing the active ingredient I would be very careful in coming to a conclusion based upon a negative finding by EDS.
FYI Polarized light microscopy can also be helpful with this type of investigation.
Regards, jr
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Wednesday, March 12, 2003 8:51 PM To: microscopy-at-sparc5.microscopy.com
On Tuesday, March 11, 2003, at 06:16 PM, Gary Gaugler wrote:
} This is perhaps a bit unusual of a topic } but it is centered on EDS viability for } foiling double blind testing. } } Question: Can EDS be used to analyze } a double blind tablet to ascertain whether } it is placebo or a study medication? } } It would seem that the inability to see } light element H might not be a limiting } factor. Are there signatures of placebos } that would strongly differentiate them } from study meds? How might one guard } against educated analysis of study meds? } If the basic elements are C, H and O, are } there distinct signatures or quantities } that one could use to distinguish between } placebo and study meds? } Dear Gary, Most medications consist of small amounts of the active ingredient mixed with a much larger amount of matrix (which provides sufficient volume to handle a tablet, gets the active ingredient into the stomach--and sometimes further along the digestive tract--dissolves at a rate appropriate to deliver the drug as desired, etc.). The placebo in a properly designed trial consists of matrix identical to that for the drug, so EDS analysis would almost certainly give the same result for both. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
We have a pleasure to announce the final circular for the:
12th Croatian Slovenian Crystallographic Meeting to be held in Hotel Jezero, National Park Plitvice Lakes, Croatia (June 19-22, 2003). All additional information with Instructions for Abstract, Registration Form and Hotel Accommodation is contained on the web site:
http://www.chem.pmf.hr/~hkz/plitvice
Looking forward to seeing you on Plitvice Lakes Organizing Committee
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (didier.goux-at-unicaen.fr) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, March 13, 2003 at 08:01:06 ---------------------------------------------------------------------------
We use a balzers CPD 020 for critical point drying. When the engineer show me how to use it, I noticed that the temperature were set to 42 degres celcius (314 K) but the critical temperature is 31 degres celcius (304 K).
Dear All This is one of those "odd ones". Seems like administration love there admin tools. Here is one they would like a realistic answer of. "How many publications can the University expect per year from a EMU with one TEM, one SEM, and one CLSM ?" The EMU is staffed by 3 people. The size of the University: Overall total 12,286 students
Full-time 9977 Part-time 2309
Male - 6328 Female - 5958
Undergraduate - 11,336 Post graduate - 950
After a nice laugh, please help me on this one. I anticipate the normal "next" question in the near future: Cost recovery through consultancy!
Stephan H Coetzee Department of Physics EMU Private Bag 0704 Abalone, Botswana
It is good to know that administrators the world over all ask for wacky things. It helps to show there are no real differences between us all. This is a silly exercise but here is how I would answer it. A realistic level of production for a faculty member is 2 refereed papers a year (clearly 5-6 would be an excellent but not impossible goal but on average, 2 would be good for cell biologists). I would suggest that you estimate 2 / year for every faculty member whose research publications would always include at least 1 micrograph and a lesser fraction for the heathen biochemist and molecular biologists who only occasionally dabble in the most difficult science of microscopy. The real flaw with this exercise is that it will be taken as a measure of productivity of the EMU when, unless your staff are terribly incompetent (an unlikely prospect), it is really a measure of faculty productivity and quality. You could have a great EMU but if you don't have aggressive faculty, it won't get used.
At 05:28 PM 3/13/2003 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
(1) Thanks to all those who replied to my 'adhesive' question. I also now remember the correspondence on this listerver concerning the sawing up of M-Bond 610 chunks. I've found that M-Bond 610 is supplied in something like 25 ml bottles - does one have to use the whole of one pair of bottles once it's opened?
(2) Something completely different - has anyone recently purchased a new vacuum coating unit for TEM? How much did it cost, and do you like it? Trying to get a price from a manufacturer directly can be very wearing - they seem to want to interrogate you about what you want to do with it, rather than letting you know if the thing is within your budget.
LAB TALK (Caption searching for a cartoon):
"Call me an airhead? There's a Penning gauge on yours!"
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
Hi All, Please allow me to pick your collective brains.... A client just requested a TEM study of mouse ankle joints. I have looked at many things in the scope over the years, but I've never had to process & cut bone. Mice have tiny bones, and this will be the ankle...even tinier: Do I need to decalcify? How should we fix (I usually use a modified Karnovsky's (2.5% GA, 4% PFA + 0,02% picric acid) as my primary fix.) How & when do I decalcify? Any preferred resin? I feel like a babe in the woods. Thanks, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I am in the market for an osmometer. Does anyone have experience with these instruments to share? I am also interested in literature from vendors. Thank you in advance.
I have grown weary of snagging digital images with a camera held in my bare hand over the eyepiece or camera port of a stereomicroscope while manipulating an object with the other hand, which actually does work passably well because of the effectively high "film" speed of the digital camera. The images have vignetting and there's usually way too much light, though. It's not a good optical match, because the digital camera's lens gets in the way; there's a need for a transfer lens ... which would be too dear and too specialized.
Therefore, I bought a suitable video adapter with C mount for my company's stereomicroscope and then bought a Kodak MDS 100 camera, both on eBay, in my price range (less than $500 in it so far). While waiting for the camera to arrive (hopefully complete in its original wrapping as advertised by the eBay seller, as Kodak has disowned the thing) I'm anticipating with the following questions:
1. Have any of you light microscopists got experiences to share ? 2. Has anyone tried to mate the camera with a PC running Linux ? 3. Are there any wavelength issues, such as poor image quality due to infrared seeping through all that glass ?
I do have experience operating a flatbed scanner that uses the USB port as well as the TWAIN interface - and that is all favorable so far. My digital camera uses floppy disks, so image transfer via that route isn't very fast, but it's pretty reliable except when I make images on very hot days, whereupon some of the floppy files are DOA. And no, I'm not trying to get it to work from a Linux PC right from the get-go. The main PC is using W98SE.
And for the time being I have given up all hope of making one of the old, defunct flatbed scanners work like a film-plane scanner. Too many mechanical and optical issues. Thanks for your help on that.
Best regards, George Langford, Sc.D. Principal Consultant Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
*From: "Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} *To: "Listserver Microscopy (E-mail)" {Microscopy-at-sparc5.microscopy.com} *Subject: Number of publications supported by the EMU per year? *Date sent: Thu, 13 Mar 2003 17:28:58 +0200
*------------------------------------------------------------------------ *The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Best regards,
Witold Zielinski
*Dear All *This is one of those "odd ones". Seems like administration love there admin *tools. Here is one they would like a realistic answer of. *"How many publications can the University expect per year from a EMU with *one TEM, one SEM, and one CLSM ?" *The EMU is staffed by 3 people. *The size of the University: *Overall total 12,286 students * *Full-time 9977 *Part-time 2309 * *Male - 6328 *Female - 5958 * *Undergraduate - 11,336 *Post graduate - 950 * *After a nice laugh, please help me on this one. I anticipate the normal *"next" question in the near future: Cost recovery through consultancy! * * *Stephan H Coetzee *Department of Physics *EMU *Private Bag 0704 *Abalone, *Botswana * *Coetzees-at-mopipi.ub.bw {mailto:Coetzees-at-mopipi.ub.bw} * *Telephone: (+267) 355 2462 *Fax: (+267) 3185 097 *Telephone: (+267) 355 0000 Switchboard * * * * :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) ;)
Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
I will preface my remarks by stating that I have not worked with osmometers since the 1980's. That said, there are two different types of osmometers/two different ways of measuring osmolarity. 1. Freezing point depression 2. Vapor pressure. How #1 works is obvious, how #2 works I have no clue. I prefered a Wescor Vapor pressuer osmometer because I could use a sample as small as 7 microliters. The freezing point depression osmometers I had access to needed a much larger volume for accurate results. That may have changed in subsequent years. Most renal physiologists have an osmometer for measuring urinc concentration, perhaps someone in the physiology dept can help.
Geoff
kwalters 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 in the market for an osmometer. Does anyone have experience with these } instruments to share? I am also interested in literature from vendors. } Thank you in advance. } } Kathy } } Kathy Walters // } Central Microscopy Research Facility / / } 85 EMRB / /\ } University of Iowa / /\ \ } Iowa City, Iowa 52242 / / \ \ } Phone #: (319) 335-8142 / / \ \ } Fax #: (319) 384-4469 ______ ((0)) } email: Katherine-Walters-at-uiowa.edu |__| / / } || / / } -------------- } ------------------
-- ********************************************** 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 mcauliff-at-umdnj.edu **********************************************
} Does anyone have experience with these instruments to share?
There are two basic types commonly used: vapor pressure osmometers and freezing point osmometers. The major manufacturer of freezing point osmometers went out of business several years ago. I am not aware of other manufactures. As a cell physiologist who works with osmoregulation, I use the vapor pressure osmometer regularly. Every colleague that I know uses the Wescor brand (Utah). I am not familiar with any other brands. I am absolutley satisfied with my Wescor, but have no means for making a comparison with other brands. These are not that cheap--I think one could cost $4-5K.
Best,
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
Greetings: I have not heard much lately about Holographic TEM lately.
From what I read, even though unstained polymer sections provide little modulation of the electron wave amplitude, there are variations in the wave phase. Holographic imaging techniques can be used to recover these phase modulations and thereby create contrast between multi-phase polymers without the use of stains. Can anyone provide an update on the state-of-the-art of this technique?
regards,
Anthony J. Ribaudo Staff Scientist Analytical Sciences Microscopy Facility Honeywell International Specialty Materials Division
Following the sudden death of my husband General Sani Abacha the former head of state of Nigeria in August 1998, I have been thrown into a state of utter confusion, frustration and hopelessness by the present civilian administration, I have been subjected to physical and psychological torture by the security agents in the country. My son is still under detention arraigned before the federal high courtof Nigeria for an offence he did not commit even though the government might release him soon. As a widow that is so traumatized,I have lost confidence with anybody within the country. You must have heard over the media reports and the internet on the recovery of various huge sums of money deposited by my husband in different security firms abroad, Some companies willingly gave up their secrets and disclosed our money confidently lodged there or many outright blackmail. In fact the total sum discovered by the Government so far is in the tune of $700. Million dollars. And they are not relenting to make me poor for life. I got your contacts through my personal research,and out of desperation decided to reach you through this medium.I will give you more information as to this regard as soon as you reply. I repose great confidence in you hence my approachto you due to security network placed on my day to day affairs I cannot afford to visit the embassy so that is why I decided to contact you and Ihope you will not betray my confidence in you. I have deposited the sumof 40 million dollars with a security firm abroad whose name is witheld for now until we open communication.shall be grateful if you could receive this fund into your account for safe keeping. This arrangement is known to you and my son Mustapha alone, so my son will deal directly with you as security is up my whole being. I am seriously considering to settle down abroad in a friendly atmosphere like yours as soon as this fund get into your account so that I can start all over again if only you wish, but if it is impossible,just help me in diverting this fund into your account which will accrue you 30% of this fund . Please honesty is the watch word in this transaction. I will require your telephone and fax numbers so that we can commence communication immediately and I will give you a more detailed picture of things. In case you dont accept please do not let me out to the security as I am giving you this information in total trust and confidence I will greatly appreciate if you accept my proposal in good faith. Please expedite action.
Sincerely yours,
Hajia Mariam Abacha.
N/B: Please copy your response:hajiamahaaba-at-netscape.net
Following the sudden death of my husband General Sani Abacha the former head of state of Nigeria in August 1998, I have been thrown into a state of utter confusion, frustration and hopelessness by the present civilian administration, I have been subjected to physical and psychological torture by the security agents in the country. My son is still under detention arraigned before the federal high courtof Nigeria for an offence he did not commit even though the government might release him soon. As a widow that is so traumatized,I have lost confidence with anybody within the country. You must have heard over the media reports and the internet on the recovery of various huge sums of money deposited by my husband in different security firms abroad, Some companies willingly gave up their secrets and disclosed our money confidently lodged there or many outright blackmail. In fact the total sum discovered by the Government so far is in the tune of $700. Million dollars. And they are not relenting to make me poor for life. I got your contacts through my personal research,and out of desperation decided to reach you through this medium.I will give you more information as to this regard as soon as you reply. I repose great confidence in you hence my approachto you due to security network placed on my day to day affairs I cannot afford to visit the embassy so that is why I decided to contact you and Ihope you will not betray my confidence in you. I have deposited the sumof 40 million dollars with a security firm abroad whose name is witheld for now until we open communication.shall be grateful if you could receive this fund into your account for safe keeping. This arrangement is known to you and my son Mustapha alone, so my son will deal directly with you as security is up my whole being. I am seriously considering to settle down abroad in a friendly atmosphere like yours as soon as this fund get into your account so that I can start all over again if only you wish, but if it is impossible,just help me in diverting this fund into your account which will accrue you 30% of this fund . Please honesty is the watch word in this transaction. I will require your telephone and fax numbers so that we can commence communication immediately and I will give you a more detailed picture of things. In case you dont accept please do not let me out to the security as I am giving you this information in total trust and confidence I will greatly appreciate if you accept my proposal in good faith. Please expedite action.
Sincerely yours,
Hajia Mariam Abacha.
N/B: Please copy your response:hajiamahaaba-at-netscape.net
On Thursday, March 13, 2003, at 08:55 AM, Robert H. Olley wrote:
} (2) Something completely different - has anyone recently purchased a } new } vacuum coating unit for TEM? How much did it cost, and do you like it? } Trying to get a price from a manufacturer directly can be very wearing } - } they seem to want to interrogate you about what you want to do with it, } rather than letting you know if the thing is within your budget. } } Dear Robert, I recently priced out several vacuum evaporators, and the one we ordered was slightly under $20k. We wanted a desk-top unit, and we wanted a turbo pump for the vacuum. It has not yet arrived, so I can't tell you whether we'll like it, but I've had good experiences with similar units. If your requirements are different, you could possibly get away with a much cheaper unit, ~$5k, with a mechanical pump for the vacuum. We need to evaporate both carbon and metals, but we don't need sputtering capability. Since the uses to which the unit will be put make a big difference in the price, I think the manufacturer or supplier is justified in asking rather than just selling the fanciest unit to each customer. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Hi Stephan This is a question of particular interest to us as we have to be accountable too and I am looking forward to the replies We use:
Utilization of our facility is monitored by 1) The numbers of researchers using the facility from individual departments. 2) The numbers of researchers using the facility from off campus and from industry. 3) The numbers of participants in the in-house workshops. 4) The numbers of graduate students taking credit courses in electron and light microscopy. 5) The numbers of students in various UBC courses visiting the facility. 6) The numbers of research papers produced with the assistance of the facility. 7) The numbers of projects handled by the facility's staff. 8) The number of research theses produced with support from the facility.
The numbers of users we mostly generate from the billing. For the numbers of papers and theses we have to remind users constantly that we need those numbers. There is a note on the billing when it is sent out. Does anyone have any other ideas how to generate these numbers? Elaine
} Dear All } This is one of those "odd ones". Seems like administration love there admin } tools. Here is one they would like a realistic answer of. } "How many publications can the University expect per year from a EMU with } one TEM, one SEM, and one CLSM ?" } The EMU is staffed by 3 people. } The size of the University: } Overall total 12,286 students } } Full-time 9977 } Part-time 2309 } } Male - 6328 } Female - 5958 } } Undergraduate - 11,336 } Post graduate - 950 } } After a nice laugh, please help me on this one. I anticipate the normal } "next" question in the near future: Cost recovery through consultancy! } } } Stephan H Coetzee } Department of Physics } EMU } Private Bag 0704 } Abalone, } Botswana } } Coetzees-at-mopipi.ub.bw {mailto:Coetzees-at-mopipi.ub.bw} } } Telephone: (+267) 355 2462 } Fax: (+267) 3185 097 } Telephone: (+267) 355 0000 Switchboard
-- Dr. Elaine Humphrey Director, BioImaging Facility First Vice President, Microscopy Society of Canada 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-interchange.ubc.ca website: www.emlab.ubc.ca
In brief, one usually sets the temperature slightly higher than the critical point to insure that one has passed it.
Cheers,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging
Mail to: Geology, CASI West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383
For help and information only, The CASI houses: An FEI Quanta 400 and Technai 12T, Oxford INCA Energy 400, Tousimis AutoSamdri 815 and Olympus FV-300.
-----Original Message----- } From: didier.goux-at-unicaen.fr [mailto:didier.goux-at-unicaen.fr] Sent: Thursday, March 13, 2003 10:23 AM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (didier.goux-at-unicaen.fr) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, March 13, 2003 at 08:01:06 ---------------------------------------------------------------------------
We use a balzers CPD 020 for critical point drying. When the engineer show me how to use it, I noticed that the temperature were set to 42 degres celcius (314 K) but the critical temperature is 31 degres celcius (304 K).
I am not sure if my first posting got through, but, the definitive answer to this question comes from either Raman microscopy or FT-IR microscopy. No guessing, just good chemical fingerprinting.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 08:53 AM 3/13/03 -0500, jrobson-at-rdg.boehringer-ingelheim.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Near IR (NIR) focal plane array systems provide direct visual differentiation of active and placebo tablets, as well as the areal distribution of components within the tablet. For an example, see http://www.spectraldimensions.com/products/index.html.
James Martin Orion Analytical, LLC www.orionanalytical.com
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Hi,
I am not sure if my first posting got through, but, the definitive answer to this question comes from either Raman microscopy or FT-IR microscopy. No guessing, just good chemical fingerprinting.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 08:53 AM 3/13/03 -0500, jrobson-at-rdg.boehringer-ingelheim.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (henryp-at-bhphoto.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, March 14, 2003 at 12:11:56 ---------------------------------------------------------------------------
Email: henryp-at-bhphoto.com Name: Henry Posner
Organization: B&H
Education: Undergraduate College
Location: New York, NY USA
Question: I am in possession of my father's WW II era Leica microscope. It's missing the chrome clips which hold a slide in place. Where can I get a pair and where can I learn more about this instrument and its current value? TIA
the company "Knauer" in Berlin / Germany builds among other scientific instruments osmometers for over 40 years. I have a freezing point one and am very contented with it. they have represantatives all over the world, just check their website http://www.knauer.net/ peter -- ********************************** peter.heimann-at-uni-bielefeld.de Dr. Peter Heimann Developmental Biology & Molecular Pathology; W7-107 University of Bielefeld D 33501 Bielefeld / Germany phone: xx49(0)521-106-5628 / 5627 FAX : " " - 5654 www.uni-bielefeld.de/biologie/Entwicklungsbiologie/ www.uni-bielefeld.de/SFB549 ***********************************
In the past I have been working on Calcium ratio measurements in microscopy. It has been a while since I was actively involved and I am curious about new developments in the field.
People have been using ultrafast camera's for Calcium ratio imaging, which puzzles me a bit. From what I remember, at 20 deg. Celsius, the Calcium sensitive probe Fura-2 needs 5-10 ms to reach equilibrium in a solution of about 140 mM Calcium. We used regular PAL video cameras (25 fps, 40 msec./frame) with a frame splitter device, which enabled us to monitor intracellular Calcium changes and relocation at 1/4 of the frame rate, at 10 msec in individual cells. Taking the Nyquist sampling theorem into account, this is sufficient to monitor most phenomena we were interested in.
We used Fura-2, as the use of Calcium ratio imaging which has some advances compared to non-ratio Calcium measurement (i.e. concentration changes of the probe). What are good alternatives to Fura-2 for intracellular Calcium measurements. For Calcium ratio measurements, a Mercury arc lamp provides the best result I guess, with its high emission of light in the near-UV range (I do not like Mercury arcs for other fluorescent work, due to their "spiky" spectrum, but prefer Xenon arcs instead, as their emission is more evenly spread through the visible spectrum).
For the optics we used Fluorite lenses, as they transmit better in the near-UV range, than glass objectives. What is your opinion of the optics for Calcium ratio measurements ?
We used mechanical filter changers for switching between the excitation wavelengths, what about elctronic devices ? From what I remember, their transmission efficiency is much lower than "traditional" filters ?
Some data : The concentration of intracellular Calcium lies between 10 nM and 10 uM or even higher I believe. The excitation wavelength optimum changes when Fura-2 binds Calcium. Fura-2 has a Kd of about 145 nM and is about saturated above 1 uM Calcium (this may change depending on the conditions ?).
When Fura-2 binds Calcium, the excitation optimum shifts between 300 and 400 nm, 363 (ion free) and 335 nm, in practice 340 nm and 380 nm are taken for the two excitation wavelengths (Mercury arc peaks). The more Calcium is present, the higher the absorption shift towards 340 nm. One can detect this shift by monitoring the emission at 510 nm (~green light), the change in emission intensity will reflect the absorption shift between 340 and 380 nm. The Kd (dissociation constant) for Calcium of Fura-2 is ~135 nM in Mg2+ free Calcium buffers and ~224 nM in the presence of 1 mM Mg2+. Although Fura-2 may accurately indicate peaks of Calcium up to 500 nM in cells, Fura-2 exhibits limited sensitivity above 500 nM Calcium.
There is no Calcium dependent change in absorption at about 360 nm., which is called the isosbestic point of Fura-2. Measurement of the 510 nm emission when exciting at 360 nm can be used as a measure for fotobleaching of Fura-2.
Dear all, I am sure I remember that someone had produced a routine for the rotational averaging of SAD ring patterns to produce an intensity versus line plot analagous to a XRD diffractogram. Could anyone give me any hints where I could get such a thing? A plug-in for Gatan's digital micrograph would be especially good, since we use this program often. A Photoshop plug-in would be another good solution.
Thanks in advance for any help you can give.
-- Ian MacLaren Technische Universität Darmstadt Material-und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany
In a message dated 3/17/03 4:48:38 AM, maclaren-at-tu-darmstadt.de writes:
} Dear all, } } I am sure I remember that someone had produced a routine for the rotational } averaging of SAD ring patterns to produce an intensity versus line plot } analagous to a XRD diffractogram. Could anyone give me any hints where } I could get such a thing? A plug-in for Gatan's digital micrograph would } be especially good, since we use this program often. A Photoshop plug-in } would be another good solution. } The Image Processing Tool Kit (for 8 bit images) and Fovea Pro (for 16 bit images) both include that routine, as a Photoshop-compatible plugin. see http://ReindeerGraphics.com
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In a message dated 3/17/03 4:48:38 AM, maclaren-at-tu-darmstadt.de writes:
} Dear all, } } I am sure I remember that someone had produced a routine for the rotational } averaging of SAD ring patterns to produce an intensity versus line plot } analagous to a XRD diffractogram. Could anyone give me any hints where } I could get such a thing? A plug-in for Gatan's digital micrograph would } be especially good, since we use this program often. A Photoshop plug-in } would be another good solution. } The Image Processing Tool Kit (for 8 bit images) and Fovea Pro (for 16 bit images) both include that routine, as a Photoshop-compatible plugin. see http://ReindeerGraphics.com
A one-year postdoctoral position (with a possible one-year extension) is opened at the CNRS in Strasbourg, France (CEPE: Centre d’Ecologie et de Physiologie Energétiques). The aim of the project is to look at the morphological and functional flexibility of the intestinal mucosa in vertebrates through light, transmission and scanning electron microscopy. An experienced microscopist is required with skills in immunohistochemistry (immunofluorescence) and transmission and scanning electron microscopy (immunogold). Skills in Environmental Scanning Electron Microscopy (ESEM) -and possibly in western blotting- would be an advantage. If interested, please send a resume to Dr JH Lignot by fax or by email.
Dr Jean-Hervé Lignot, Université Louis Pasteur / CNRS CEPE 23 rue Becquerel, 67087 Strasbourg, Cedex 2, France Fax: 00 33 0388106906 Tel: 00 33 0388106938
A one-year postdoctoral position (with a possible one-year extension) is opened at the CNRS in Strasbourg, France (CEPE: Centre d’Ecologie et de Physiologie Energétiques). The aim of the project is to look at the morphological and functional flexibility of the intestinal mucosa in vertebrates through light, transmission and scanning electron microscopy. A good microscopist is required with skills immunohistochemistry (immunofluorescence), transmission and scanning electron microscopy (immunogold). Skills in Environmental Scanning Electron Microscopy (ESEM) -and possibly in western blotting- would be an advantage. If interested, please send a resume to Dr JH Lignot by fax (0033 388106 906) or by email (J-H.Lignot-at-c-strasbourg.fr).
Dr Jean-Hervé Lignot, Université Louis Pasteur / CNRS CEPE 23 rue Becquerel, 67087 Strasbourg, Cedex 2, France Fax: 00 33 0388106906 Tel: 00 33 0388106938
I'm building a humidified/heated chamber to do live cell imaging of mammalian cells. iIm looking for a vendor that sells warm air blowers that would be useful.
Thanks in advance for the input.
Best,
Gary
Gary S. Laevsky, Ph.D. Research Associate The Scripps Research Institute 10550 N. Torrey Pines Road/IMM-24 La Jolla, CA 92037 (858) 784-9372
I am in the market for a electropolisher for TEM sample preparation & was wondering if anyone had any suggestions. Preferably a twin jet electropolisher for amorphous metal sample preparation.
Any feedback would be greatly appreciated,
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
Does anyone know where I could get another rubber drive belt for the Reichert OMU3 ultramicrotome? Ours broke, and when we try to glue it, it just keeps breaking again.
Does this rotational averaging routine from Reindeergraphics take into account the ellipticity of the (imperfect) SAD ring patterns? If so, does it automatically find the major and minor axes or does that require manual adjustment?
Thanks,
Tom Schamp
On 3/17/03 7:03 AM, ""DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com" {"DrJohnRuss-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 } -----------------------------------------------------------------------. } } } } In a message dated 3/17/03 4:48:38 AM, maclaren-at-tu-darmstadt.de writes: } } } Dear all, } } } } I am sure I remember that someone had produced a routine for the rotational } } averaging of SAD ring patterns to produce an intensity versus line plot } } analagous to a XRD diffractogram. Could anyone give me any hints where } } I could get such a thing? A plug-in for Gatan's digital micrograph would } } be especially good, since we use this program often. A Photoshop plug-in } } would be another good solution. } } } The Image Processing Tool Kit (for 8 bit images) and Fovea Pro (for 16 bit } images) both include that routine, as a Photoshop-compatible plugin. see } http://ReindeerGraphics.com } } } }
---------------- Tom Schamp ph: (434) 982-4595 University of Virginia fx: (434) 982-5660 Dept. of Materials Science and Engineering email: cts2v-at-virginia.edu 116 Engineer's Way Charlottesville, VA 22904
I am interested in looking at the general Laminin distribution within a basement membrane. Does anybody know of, or have, an antibody that will cross react with a number of Laminin subclasses, rather than just a specific Laminin subclass ?
I am more than happy to receive answers from antibody suppliers.
Thanks in advance
Allan
-- ------------------------------------------------- Allan Mitchell Technical Manager Otago Centre for Electron Microscopy C/-Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
On Monday, March 17, 2003, at 01:40 AM, Ian MacLaren wrote:
} I am sure I remember that someone had produced a routine for the } rotational averaging of SAD ring patterns to produce an intensity } versus line plot analagous to a XRD diffractogram. Could anyone give } me any hints where I could get such a thing? A plug-in for Gatan's } digital micrograph would be especially good, since we use this program } often. A Photoshop plug-in would be another good solution. } } Thanks in advance for any help you can give. } Dear Ian, I wrote a routine both for SPIDER and as a stand-alone, which determines the center and radius of a ring from an input of between 3 and 20 points on the ring. SPIDER can then produce a rotational average by padding your original diffractogram into a larger image such that the center of the pattern is the center of the larger image and using the rotational averaging module in SPIDER. This does not qualify as a plug-in for DM, but it could do the job. Since I have moved from Albany, I no longer have access to the code, but perhaps someone there could get it to you, if you decide you need it. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Ian, You may want to look at the program ProcessDiffraction http://www.mfa.kfki.hu/~labar/ProcDif.htm by János L. Lábár. It will do what you are asking and is a stand alone program. I have used it a handful of times, but never very intensely.
-Ray
Ray D. Twesten, PhD. Center for Microanalysis of Materials Seitz Materials Research Laboratory 104 S. Goodwin Ave., Urbana, IL 61801 +1 217 244-6177 (Fax -2278)
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: Monday, March 17, 2003 3:41 AM To: Microscopy Listserver Cc: Ralf Theissmann
Dear all, I am sure I remember that someone had produced a routine for the rotational averaging of SAD ring patterns to produce an intensity versus line plot analagous to a XRD diffractogram. Could anyone give me any hints where I could get such a thing? A plug-in for Gatan's digital micrograph would be especially good, since we use this program often. A Photoshop plug-in would be another good solution.
Thanks in advance for any help you can give.
-- Ian MacLaren Technische Universität Darmstadt Material-und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany
Rotational averaging is included in a software package for Digital Micrograph written at the NCEM. It's available for download at http://ncem.lbl.gov/frames/software.htm.
Larry Thomas Pacific Northwest National Laboratory P.O. Box 999 Mail Stop P8-16 Richland, WA, USA
} ---------- } From: Tom Schamp } Sent: Monday, March 17, 2003 11:13 AM } To: "DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com; ian.maclaren-at-physics.org; Microscopy-at-sparc5.microscopy.com } Subject: Re: Rotational averaging } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dr. Russ, } } Does this rotational averaging routine from Reindeergraphics take into } account the ellipticity of the (imperfect) SAD ring patterns? If so, does } it automatically find the major and minor axes or does that require manual } adjustment? } } Thanks, } } Tom Schamp } } } On 3/17/03 7:03 AM, ""DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com" } {"DrJohnRuss-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 } } -----------------------------------------------------------------------. } } } } } } } } In a message dated 3/17/03 4:48:38 AM, maclaren-at-tu-darmstadt.de writes: } } } } } Dear all, } } } } } } I am sure I remember that someone had produced a routine for the rotational } } } averaging of SAD ring patterns to produce an intensity versus line plot } } } analagous to a XRD diffractogram. Could anyone give me any hints where } } } I could get such a thing? A plug-in for Gatan's digital micrograph would } } } be especially good, since we use this program often. A Photoshop plug-in } } } would be another good solution. } } } } } The Image Processing Tool Kit (for 8 bit images) and Fovea Pro (for 16 bit } } images) both include that routine, as a Photoshop-compatible plugin. see } } http://ReindeerGraphics.com } } } } } } } } } } } } } ---------------- } Tom Schamp ph: (434) 982-4595 } University of Virginia fx: (434) 982-5660 } Dept. of Materials Science and Engineering email: cts2v-at-virginia.edu } 116 Engineer's Way } Charlottesville, VA 22904 } } } } }
This is an open reply to the listserver for your posting, but my questions are directed to you.
I tried the radial profile option in Fovea Pro on a ring pattern file that I have that has the pattern centered. I got a nice radial profile. I then offset the image and put a black background where the image was offset. If you take that radial profile, you get something quite different. Could you tell us the general algorithm that you use to obtain this?
I would also like to know whether the circular Hough Transform that you discussed with me a few years ago would take into account this offset or must the pattern be centered?
Again with the circular Hough Transform, could that address an elliptical pattern?
On another note:
I helped MDI, Inc. modify their electron diffraction module in WinJade several years ago. Here are some of the problems that had to be addressed with their system. 1) some diffraction patterns can have an aspect ratio that is not 1:1 either due to the microscope or the digitizing process. This needs to be taken care of because it can be as much as 2%. For the microscope itself, the presence of an elliptical pattern can be checked quite easily by taking two identical diffraction patterns of polycrystalline gold and overlaying one over the other, but rotated 90 degrees. 2) patterns are often off-center and must be centered for the radial profile 3) Their original algorithm simply took the number of pixels in a ring and divided by the number of pixels in the ring for an average. This had the effect of decreasing the intensities quickly. In effect, they were taking the integral over dxdy, not the integral over d(theta)rdr, i.e. not taking the average in polar coordinates. For spotty patterns, this drastically cut down the intensities at larger radii in the pattern because of the large number of black pixels in the rings bring the intensities down. We developed four algorithms for using on patterns and which was best depended on the type of pattern e.g. completer rings, spotty rings, etc. The four algorithms were:
J1: Average All Pixels in a Ring --The radius for each pixel in the image is calculated, the intensity at each radius is summed, divided by the number of pixels at that radius, and the pattern is normalized to 10,000 counts.
J2: Average Above-Average Pixels --The average of each radius is found and only pixels at each radius that have intensities above the average for the radius is used. The intensity is calculated for each radius by averaging these pixels and the pattern is normalized.
J3: Sum of Above-Average Pixels --The sum of the above-average pixels is used instead of the average and the pattern is normalized.
J4: Take Maximum Pixel in Ring --The highest intensity pixel in each ring is used as the intensity.
Here are the conclusions that I found with WinJade to real world diffraction patterns:
Care must be taken when choosing the mode of data reduction. J1 and J2 give very good results with complete rings. J2 works very well with incomplete but readily apparent rings. J2 gives an improved peak/bkg ratio because it excludes pixels below the average intensity value for a given ring. J3 is noisy. It works well when there are discrete spots on a black background. J4 works well for spotty patterns and finds all of the d-spacings present. It provides good values of d-spacings at large radii.
This program was really great for working with SAD patterns. You could compare the radial profile with the powder diffraction database, compare with X-ray diffraction data, overlay either experimental XRD data or PDF patterns on the SAD pattern, and easily calibrate the camera constant easily for the program. Unfortunately, in the new version of WinJade, they did not include the electron diffraction module. They have it in another program that they sell, but I already bought the upgrade to the new program and can't use the old version on Windows 2000. -Bummer.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: "DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com [mailto:"DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com] Sent: Monday, March 17, 2003 7:04 AM To: ian.maclaren-at-physics.org; Microscopy-at-sparc5.microscopy.com
In a message dated 3/17/03 4:48:38 AM, maclaren-at-tu-darmstadt.de writes:
} Dear all, } } I am sure I remember that someone had produced a routine for the rotational } averaging of SAD ring patterns to produce an intensity versus line plot } analagous to a XRD diffractogram. Could anyone give me any hints where } I could get such a thing? A plug-in for Gatan's digital micrograph would } be especially good, since we use this program often. A Photoshop plug-in } would be another good solution. } The Image Processing Tool Kit (for 8 bit images) and Fovea Pro (for 16 bit images) both include that routine, as a Photoshop-compatible plugin. see http://ReindeerGraphics.com
Scott (and anyone else interested in this conversation)
The plugin uses a circular hough to get the radially averaged intensities. It expects the center of the pattern to be "near" the center of the image, as it does not store the full 3D accumulator space for a Hough (the axes are x-center, y-center and radius). I suppose that with computer memories getting larger it would be possible to store a larger array, if someone has a pressing need. The method works by adding up the grey scale brightnesses of each pixel in the original image into the points in the Hough array that define a possible circle through those points - in other words a cone in the Hough space. Then the central axis of the pattern is found by locating the brightest column in the space, and the intensities along that axis form a plot of the circularly averaged intensities. The routine does NOT perform an elliptical hough transform, which would require a 5 dimensional array (x-center, y-center, minor axis, major axis, angle). It is certainly possible to write something to do that, but it simply hasn't come up before. What I usually do with images that have some distortion (e.g., non-round SAED patterns) is fix that first, before the measurement. But writing an elliptical Hough wouldn't be hard (given enough memory) and if someone wants to agitate for that, I'll consider doing it. Might even give it away, as this seems like a pretty specialized application.
John Russ
In a message dated 3/17/03 8:05:29 PM, walck-at-ppg.com writes:
I tried the radial profile option in Fovea Pro on a ring pattern file that I have that has the pattern centered. I got a nice radial profile. I then offset the image and put a black background where the image was offset. If you take that radial profile, you get something quite different. Could you tell us the general algorithm that you use to obtain this?
I would also like to know whether the circular Hough Transform that you discussed with me a few years ago would take into account this offset or must the pattern be centered?
Again with the circular Hough Transform, could that address an elliptical pattern?
No question it can be done. All of the math is pretty trivial, but there is a modest amount of programming involved and before embarking on it I'd like to see how much interest there is. Apparently someone has written something that does circular averaging in DM, and if that meets peoples' needs there is no point in my writing a Photoshop plugin as well. If a groundswell of interest develops (i.e., more than just you), I'll try to design a fairly simple but adequate user interface to get the camera constant, so that the output plot and Excel file have all of the 1/d, d, etc values included. If the ellipticity of the pattern is an instrument constant, it is still easier to fix it before measurement rather than add the extra parameters to the Hough.
John Russ
=====
In a message dated 3/17/03 8:48:57 PM, walck-at-ppg.com writes:
} John, } } I think that you would be surprised how many elliptically patterns are } out there that people don't even know they have until someone points it } out. The microscope manufacturers don't go out of their way to tell you. } } } } One more question for you. How do you calibrate the output. The data } is given (in my case) with a maximum radius as something like 4.6 um. } I then went in and made the um/pixel value =1 and so now even though it } is saying that the max radius is in um, I know that it is in pixels. Still, } is there anyway you could have a diffraction pattern calibration in the } program similar to the magnification calibration for images? this way, } d-spacing, 1/d spacings, 2theta, values, and 2theta(XRD Cu-Ka) values could } be calculated. } } } } I could see an elliptical check with the camera constant calibration. } Two vertical lines that you can adjust in and out so that they are tangent } to opposite sides of the same ring and then two horizontal lines to do } the same thing. The value of the separations of the lines would agree } for a circular pattern as a check. This could also check the centering } of the pattern when the two lines are over top of each other and their } separation is zero. } } } } -Scott
I was wondering if anyone can tell me where I might find demographics for research and clinical labs in the US. Any site's you can refer me to would be greatly appreciated.
*Date sent: Mon, 17 Mar 2003 10:53:06 -0600 *From: William Stratton {wgstratton-at-wisc.edu} *Subject: Recommendations for Electropolisher? *To: Microscopy-at-sparc5.microscopy.com
*------------------------------------------------------------------------ *The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
We're using double jet polisher Tenupol made by Struers for many years and may say that it's good maschine for metallic TEM specimen preparation. However such requirment as electropolishing of amporhous metal is maybe too specific. Good performence of this method depends rather on the metal than on its state.
Best regards,
Witold Zielinski
*Greetings all, * *I am in the market for a electropolisher for TEM sample preparation & was *wondering if anyone had any suggestions. Preferably a twin jet *electropolisher for amorphous metal sample preparation. * *Any feedback would be greatly appreciated, * *William Stratton * *------------------- *William G. Stratton *Research Assistant *University of Wisconsin - Madison * *1509 University Avenue *Madison, WI 53706 *Office: 608-265-6391 *Fax: 608-262-8353 *wgstratton-at-wisc.edu * * * :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) ;)
Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
The company Solent Scientific manufactures full enclosure incubation chambers for Research Inverted Microscopes, Confocal Microscopes and Multi Photon Microscopes. Maybe they can help with the necessary equipment ?
Their website: http://www.solentsci.com/
Note: I have no commercial relation with this company. At my previous employer they used to build their own fully equipped incubator chambers for long-term time-lapse studies of mammalian cells (} 24 h.).
==================== Gary Laevsky 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 building a humidified/heated chamber to do live cell imaging of } mammalian cells. iIm looking for a vendor that sells warm air blowers that } would be useful. } } Thanks in advance for the input. } } Best, } } Gary } } Gary S. Laevsky, Ph.D. } Research Associate } The Scripps Research Institute } 10550 N. Torrey Pines Road/IMM-24 } La Jolla, CA 92037 } (858) 784-9372
Dear Garry, You can get it stitched neatly, That is what I did for my Reichert ultracutE, It is working for 3 years like that. Shashi
Does anyone know where I could get another rubber drive belt for the Reichert OMU3 ultramicrotome? Ours broke, and when we try to glue it, it just keeps breaking again.
__________________________________________________ Do you Yahoo!? Yahoo! Platinum - Watch CBS' NCAA March Madness, live on your desktop! http://platinum.yahoo.com
Does anyone have a source for black or darkly colored membrane filters. I am setting up to do an OM particle counting / sizing study and need a filter with a pore size of {0.45 micrometers that will be resistant to cyclohexane.
Thanks, John A. Robson Boehringer Ingelheim Pharmaceuticals, Inc. PO Box 368 900 Ridgebury Rd Ridgefield, CT 06877
The University of Texas Health Science Center at San Antonio will host a Symposium and Workshop sponsored by Hamamatsu Photonics KK on
Fluorescence Lifetime Imaging and Spectral Imaging
Academic Participants: Philippe Bastiaens (GER)*Robert Clegg (USA)*Mary Dickinson (USA)*Paul French (UK) Hans Gerritsen (Neth)*Brian Herman (USA)*Ammasi Periasamy (USA) Herbert Schneckenburger (GER)*Ton Visser (Neth)*Timo Zimmermann (GER)
Symposium Registration Deadline: May 1, 2003 June 6-7, 2003 The Sheraton Gunter Hotel 205 E. Houston St. San Antonio, TX Student: $200 ($250 after May 1) Professional: $250 ($300 after May 1)
Workshop Application Deadline: April 7, 2003 June 8-10, 2003 UT Health Science Center San Antonio, TX Tuition: $700 (includes room and board) 20 student limit/4 scholarships
For Information and Forms Visit: http://usa.hamamatsu.com/flim_spectral/default.htm or http://www.uthscsa.edu/csb/imaging-course.html
Rushing around South Africa at the moment I have just seen the request for data on Spot Size in a TEM, perhaps I can help?
The Spot Size control adjusts the first condenser lens and in combination with the second condenser lens they demagnify the virtual source to provide a certain beam "spot size" on the specimen. This spot size may be measured by setting the magnification at 10,000X where 1um = 1cm. When trying to produce a quality image I would always suggest one uses a spot size smaller than 3um.
Whilst the Spot Size is used by most people at its largest setting (bright) it is not the way the designers expect the instrument to be. Good operating techniques should always take the spot size into account, even at lower magnifications better quality images will be attained at smaller spot sizes.
Since papers were produced in 1944 about transmission images (LM and EM) we have known that parallel beams are very important if you are chasing the best image quality, biology or materials we always desire high coherence. There are some misunderstandings on how to obtain a parallel beam or high coherence, for example setting the final condenser under focus is incorrect. The procedure for high coherence would be to use the smallest spot size you could tolerate (this probably means you must up the emission current, use at least 20 to 30 uA). Once in this condition over focus the final condenser (clockwise from crossover) the spot, whatever size it is now, becomes your new virtual source. The further over focus you go, the greater the distance between the specimen and the crossover the more parallel the beam and therefore it attains a higher coherence, which is what we are after. You will deduce the smaller the condenser aperture the sharper the spot and the smaller the spot the greater the coherence for a given degree of over focus.
Work with a design team and they expect everyone to over focus, they expect everyone to use small condenser apertures and they expect everyone to use small spot sizes. They do not expect everyone to use too low an emission current because this makes the task too difficult! Unfortunately almost everyone does use too low a current, I have talked before about filament life being the most important feature of many laboratories and it is always to the determent of image quality.
So in short you should always use a smallish spot size when taking micrographs and you should always run with the second condenser over focus. With sheet film photography try to use 3 to 4 seconds exposure, as this will give you better coherence too. Also remember that the denser the negative the more contrast you will build in your final print.
Hope this helps, spot size IS so important in the production of a quality image.
Steve Chapman
Senior Consultant EM
Protrain for Electron Microscopy Courses World Wide
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (alvarobq-at-fcien.edu.uy) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, March 18, 2003 at 11:42:39 ---------------------------------------------------------------------------
Question: may i adding potassium ferrocyanide to 1%OsO4 postfixation to enhance contrast-staining cellular components in a routin coloidal-gold immunocytochemistry method? how do you prepare this ferrocyanide solution? what rate do you use? and if i use Lowicril(absence of osmium treatment), may i adding to the glutaraldehyde solution? how do you prepare and what rate?
Can anyone fill me in on some of the issues surrounding doing high res cryoEM and/or the need for something like a PhotoScan 2002 high res scanner from ZI Imaging.
I need to get some of the basics down so I can have a reasonable chance of understanding a proposal being made here.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have a guy who wants to look at the interior of some wood blocks using our conventional SEM. He is trying to see bacteria and anything else that might be hiding in there.
His blocks are pretty small, about 5mm cubes. I thought about CPD, but had second thoughts about how long to soak in CO2 etc. He did some experiments and it took a couple of hours for some dye to make it all the way into the middle of the block. Plus he has about 30 of these things to look at. That could add up to a lot of time and CO2.
I got an idea that HMDS might work for this project, but have no experience with it. I think that being able to soak the blocks for a long time in HMDS would be good, as would the chance to let them sublime to dryness over a long time without too much attention.
Do you think I am on the right track or is there something I am missing.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
A journal editor requested me to provide a reference regarding how to measure electron dose. I looked over some textbooks and experimental sections of some papers on electron beam damage. However, I could not find. If someone knows a reference, please advise.
Hiromi Konishi University of New Mexico hkonishi-at-unm.edu
Steve, thanks for the very good comment on spot size issue.
Without knowing all these details I was told many years ago to use smaller possible spot size (yes, I do have some idea what coherence is) and owerfocus second condenser. Working in Russia and here in US I always set all my TEMs for spot size 3 (JEM12000EX) surviving the battle with previous "spot size 1" lovers. If microscope aligned correctly, I never ever had a problem with illumination even with regular tungsten filament in magnification range x5-80K. My standard exposure time is 1.5-1.7 sec for 4489 film. In order to insure dynamic range, negatives should be very grayish. If it's too dark - you will loose the film's dynamic range. If you need extra contract - use more contrasty paper. In the nowadays, when we switched mostly for digital imaging, low illumination is just not an issue. My digital camera is exactly 10x more sensitive than 4489 film, so I have exposure time 0.1-0.2 sec with tungsten filament, spot size 3. Another reason to use smaller spot size - it's more easy to focus the images, at least to me: I do see focus changes more clear with smaller spot size. As for illumination, we have to keep in mind that e-beam itself damage our fragile biological samples, so less light - better for your sample. Thanks again for such detailed comment. Sergey
At 08:20 AM 3/18/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I saw a black (Whatman) Nuclepore polycarbonate membrane filter (0.2 and 0.4 um pore) in VWR. You'd have to check the chemical resistance charts if polycarbonate is resistant to cyclohexane.
Lizette Tuason
-----Original Message----- } From: "jrobson-at-rdg.boehringer-ingelheim.com"-at-sparc5.microscopy.com [mailto:"jrobson-at-rdg.boehringer-ingelheim.com"-at-sparc5.microscopy.com] Sent: Tuesday, March 18, 2003 5:28 AM To: microscopy-at-sparc5.microscopy.com
Jonathan,
We recently looked at wood blocks in our FESEM and the only preparation we did was to freeze them in liquid nitrogen, then fracture them with pliars to expose the internal structure, followed by several days in a vacuum chamber to get as much moisture and air out as possible. We glued them down with copius amounts of conductive paint, then sputtered the bejesus out of them.
The structure was well preserved and we did find bacteria.
The only problem was breaking the harder pieces of wood. Some of the wood was ancient and very brittle, but the new pieces were tough.
Hope this is applicable to your problem.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Jon Krupp [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Tuesday, March 18, 2003 3:03 PM To: Microscopy-at-sparc5.microscopy.com
Hi again:
I have a guy who wants to look at the interior of some wood blocks using our conventional SEM. He is trying to see bacteria and anything else that might be hiding in there.
His blocks are pretty small, about 5mm cubes. I thought about CPD, but had second thoughts about how long to soak in CO2 etc. He did some experiments and it took a couple of hours for some dye to make it all the way into the middle of the block. Plus he has about 30 of these things to look at. That could add up to a lot of time and CO2.
I got an idea that HMDS might work for this project, but have no experience with it. I think that being able to soak the blocks for a long time in HMDS would be good, as would the chance to let them sublime to dryness over a long time without too much attention.
Do you think I am on the right track or is there something I am missing.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Has anybody experience in praparation of weak milk foams? We prefer freeze fracture technique. The bubbles size is at maximum 1 mm. Of special interest is the air-milk interface.
The next meeting of the Metropolitan Microscopy Society will be held on April 9th, 2003 at the EDAX facility in Mahwah, NJ. EDAX has graciously agreed to host our meeting and to provide complimentary coffee and lunch to all the attendees.
The forthcoming meeting comprises five presentations offering a blend of microscopic, spectroscopic and image processing applications. Mike Marko will describe the cutting edge methods for tomographic reconstruction of biological organelles. Paul Kotula and Tom Hancewicz will discuss methods to extract maximum value from the data by analyzing spectral images. Matt Libera will illustrate ways to form novel surface patterns on polymeric materials, using electrons, for controlling biological activity and Del Redfern will highlight the advances in high-resolution imaging in projection x-ray microscopy.
We invite all members to attend and to inform their colleagues and fellow workers who may also wish to attend the meeting. This will not only offer an opportunity to listen to some of the experts in their respective fields but also a venue to meet colleagues and exchange ideas as well as to discuss some of the new items that are in the pipeline for this year.
It is important that members should pre-register as it will help us plan for lunches. The pre-registration deadline is April 4 and can be accomplished electronically. Please respond via email or fax to Evan Slow directly. For all attendees, the meeting fee, which includes lunch, will be $20.00.
For any additional information about the meeting, please contact any of the officers. We hope to see you on April 9th at the meeting in Mahwah!
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Hi. I am looking for a reference regarding electron dose measurement. Journal editor requested me to provide a reference on dose measurement in microscope. I looked over some textbooks and papers on electron damages, but I could not find. If someone knows a reference (textbook or experimental section in papers), please advise. Thank you,
Hiromi Konishi, Ph.D. University of New Mexico hkonishi-at-unm.edu
I was wondering if anyone can tell me where I might find demographics for research and clinical labs in the US. Any site's you can refer me to would be greatly appreciated.
I'm not sure that the requirements are clearly stated but I'll try my best to respond. If the person already has the blocks of wood cut, and they are only 5mm square, then they most likely are dried out as are any bacteria within the blocks and there is no point in rehydrating and then dehydrating with HMDS or CPD - the damage has already been done. On the other hand, if the blocks of wood are in a buffer solution, then perhaps HMDS would work but you will have to soak for quite a while and then it will take some time to dry. Five mm blocks could be dehydrated in a graded ethanol series (10, 20, 30, 50, 70, 80, 95, 3x100, 3:1 ethanol:HMDS, 1:1, 1:3 and then 100% HMDS. I'm guessing on the times as it would depend on the type of wood. I'd try 30min or so for each step.
How does he plan to look at anything in the interior of the block with an SEM? Will he section them? Depending on the wood, he might need a sliding microtome and a good sharp microtome blade (I've never had much success with razor blades and always preferred a good heavy, sharp blade. If he is going to look at only the surface, then whatever is there is already dried by the above process. I really don't know how the xylem parenchyma and the xylem rays are going to look (I'm assuming secondary xylem as you wrote "wood blocks". And depending on the material and what he wants to look at he might have to embed in some type of removable embedment.
If he is planning on sectioning the wood then the blocks will have to be soaked in warm water or sometimes I've had to use steam to soften the lignified secondary walls immediately before and during sectioning. This would create sections that you could then run up into HMDS; they would dehydrate much faster.
Just a few ideas and thoughts, Damian Neuberger
} Hi again: } } I have a guy who wants to look at the interior of some wood blocks using } our conventional SEM. He is trying to see bacteria and anything else that } might be hiding in there. } } His blocks are pretty small, about 5mm cubes. I thought about CPD, but had } second thoughts about how long to soak in CO2 etc. He did some experiments } and it took a couple of hours for some dye to make it all the way into the } middle of the block. Plus he has about 30 of these things to look at. That } could add up to a lot of time and CO2. } } I got an idea that HMDS might work for this project, but have no experience } with it. I think that being able to soak the blocks for a long time in HMDS } would be good, as would the chance to let them sublime to dryness over a } long time without too much attention. } } Do you think I am on the right track or is there something I am missing. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
I would propose to measure the beam current by using a Faraday cage and, by integration in time, you can get an acceptable approximation of the electron dosis of irradiation of your sample. The Faraday cups/cages are commercially available; you may have some doubts about the precision of electron beam current measurement, but it will work.
Sincerely,
Corneliu Sarbu
At 19/03/03 13:40, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Corneliu Sarbu, PhD Department of Metallurgy and Applied Materials Science (MTM Dept.) Catholic University of Leuven (KULeuven) Kasteelpark ARENBERG nr. 44 B-3001 Heverlee-Leuven, Belgium **************************************************************** Phone: +32-16-32.1241 - office +32-16-32.1264 - secretary of department Fax: +32-16-32.1992 or +32-16-32.1270 e-mail: Corneliu.Sarbu-at-mtm.kuleuven.ac.be web-site: www.mtm.kuleuven.ac.be ****************************************************************
I would propose to measure the beam current by using a Faraday cage and, by integration in time, you can get an acceptable approximation of the electron dosis of irradiation of your sample. The Faraday cups/cages are commercially available; you may have some doubts about the precision of electron beam current measurement, but it will work.
Sincerely,
Corneliu Sarbu
At 19/03/03 13:40, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Corneliu Sarbu, PhD Department of Metallurgy and Applied Materials Science (MTM Dept.) Catholic University of Leuven (KULeuven) Kasteelpark ARENBERG nr. 44 B-3001 Heverlee-Leuven, Belgium **************************************************************** Phone: +32-16-32.1241 - office +32-16-32.1264 - secretary of department Fax: +32-16-32.1992 or +32-16-32.1270 e-mail: Corneliu.Sarbu-at-mtm.kuleuven.ac.be web-site: www.mtm.kuleuven.ac.be ****************************************************************
SEM -cryo. need info on conduction by Leit-C CCC at low temperatures
I am looking at samples of ice (glacier & artificial chemical doped) using SEM with cryo-stage.
The samples are stuck to stubs using Leit-C conductive carbon cement. A co-worker has suggested to me that carbon has greatly reduced electrical conduction at liquid nitrogen temperatures but could not remember where he had seen it. Does anyone have any information on this? Thanks,
David Mallard Department Earth Sciences Bristol University, UK
Please take note of the upcoming School on Electron Microscopy of Powdered Nanostructured Materials
which will be hold in Wroc?aw (Poland), 19-20 September 2003. For complete information, please look at: http://celtam.int.pan.wroc.pl/ISEM/ Or Email: kepinski-at-int.pan.wroc.pl
Looking forward to seeing you in Wroc?aw Organizing Committee
Currently I am working on a project which need label cytokeratin in skin sample. we use NCK-CK15 from Novocastra Lab as primary antibody. we got nice result on paraffin section at LM level. But we couldn't get any K15 gold labeling on LR white sections. The sample was fixed with 4% paraformaldehyde and embedded in LR White at 50C for 1 day. We did get gold labeling of filaggrin on the LR white sections from same block. I was wondering if the cytokeratin antigen survived after sample preparation. Here are my questions. 1) Has anyone done this kind of labeling on resin embedded sections? If anyone did similar experiment, can you share the reference or protocol? 2) Immunohistochemistry always need perform antigen retrieval on sections before labeling, what about EM immunolabeling? 3) I have some references about antigen retrieval of resin sections. I tried high pressure heat and microwave heat with sodium citrate buffer. But I lost almost all sections from grids. Is there any tricks to keep sections stick on the coated or bare nickel grids.
Any suggestions are really appreciated!
Shanling
Shanling Shi Advanced Imaging & Measurement Unilever Research & Development -Edgewater 45 River Road Edgewater, NJ 07020 201-840-2340 Shanling.Shi-at-unilever.com
The U.S. Post Office’s new Merlin (?) system has rejected the following addresses from the mailing of the March issue of Microscopy Today.
These subscribers were not sent magazines. I noticed several instances of 4-digit ZIP codes, other than that they look OK to me.
If you see your name here will you please resubscribe at http://www.microscopy-today.com
Thanks,
Ron Anderson, MT Editor
Ms. Becca Hoffman Triquint Semiconductor Dr. Stephen Giovannoni Oregon State Univ Dr. Andrew Gabor Evergreen Solar Inc Dr. Emil Adamec McLean Hospital Dr. Arthur Coates Microtherm Mr. Vincenzo Lordi Princeton University Dean Face DuPont Ms. Hermina Borgerink Wake Forest Univ Medical School Ms. M.L. Henson Usacil - Conus Mr. Larry L. Flinn US Army Criminal Inv Lab-Conus Larry Flynn Usacil-Conus[[same person]] Dr. Irene Kokkala North Georgia College Heike Gabrisch Cal Tech Ms. Vina R. Diderrich Univ Of Tennessee Ms. Judith A. Mescher Desc - Eltf Dr. Lewis D. Stegink Univ Of Iowa Mr. Wayne Engleman Monsanto BB3G Dr. John Longlet Chevron Chemical Co Daryl L Goad Texas A & M University Mr. John Curulli University of Southern California Dr. David Carter Center for Plant Cell Biology Ms. Susan DeMaggio Univ Of California Matthias Rief Stanford Univ Victoria Doyle-Jones Stanford University
Actually, this is a fairly short list for a database of over 16,500 subscribers!
Just a reminder about the exciting and innovative joint meeting of the Oklahoma Microscopy and Central States Microscopy and Microanalysis Societies that will be held at the University of Tulsa on April 10-11. Entitled "The Art of the Scientific Image", the two-day event will focus on the creation of scientific images, their digital processing and analysis, and their effective presentation. Connections between scientific imaging, human vision, and art will be explored throughout the meeting, which will feature an art exhibit and a region-wide contest for scientific images presented for aesthetic appeal rather than scientific content. Renowned imaging expert Dr. John Russ, author of The Image Processing Handbook, will present a workshop on the basics of image analysis, and Dr. Rob Weaver from the McCrone Research Institute will demonstrate techniques for forming beautiful images using specialized light microscopy techniques. A highlight of the meeting will be the keynote address by David Scharf, called by Time magazine "an Ansel Adams of inner space," whose scanning electron microscope images are regularly published in science journals, as well as in popular magazines such as Time, Nature, and Discovery. His address will accompany a viewing of the IMAX film "Hidden Dimensions" for which his contributions received an Emmy award. The meeting will close with an address by Dr. Lynn Gamwell, whose recent book Exploring the Invisible documents two centuries of connections between science, art, architecture, and culture.
The program and a registration form can be found on our website, see below
We are looking into a chartering a bus for those wishing to attend the meeting. The cost would be ~$50 per person and need to reserve it by Wednesday March 26 so please let us know asap if you are interested.
For more information contact Randy (tindallr-at-missouri.edu) or Lou (rosslm-at-missouri.edu).
Thanks, Lou Central States Microscopy and Microanalysis Society http://treefrog.cvm.uiuc.edu/~lam/csmms/
We have immunolabeled human skin for filaggrin and keratins at both light and electron microscopic levels. We have used mostly Zamboni's fixed tissue embedded in LRWhite with good success for filaggrin, K10 and Pan Keratin. However, I haven't tried K15. Two of my favorite tricks for unmasking are, 1) 0.3% sodium dodecyl sulphate (SDS) in 0.01M tris pH 7.6 incubated for 5-10 min, then rinse and continue as normal. 2) incubate the grid on 0.01% trypsin in PBS for 5-10 min., rinse and continue as normal.
Robert Underwood U of Washington Div. of Dermatolgy
On Thu, 20 Mar 2003, Shanling Shi 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. there, } } Currently I am working on a project which need label cytokeratin in skin } sample. we use NCK-CK15 from Novocastra Lab as primary antibody. we got nice } result on paraffin section at LM level. But we couldn't get any K15 gold } labeling on LR white sections. The sample was fixed with 4% paraformaldehyde } and embedded in LR White at 50C for 1 day. We did get gold labeling of } filaggrin on the LR white sections from same block. I was wondering if the } cytokeratin antigen survived after sample preparation. Here are my questions. } 1) Has anyone done this kind of labeling on resin embedded sections? If anyone } did similar experiment, can you share the reference or protocol? } 2) Immunohistochemistry always need perform antigen retrieval on sections } before labeling, what about EM immunolabeling? } 3) I have some references about antigen retrieval of resin sections. I tried } high pressure heat and microwave heat with sodium citrate buffer. But I lost } almost all sections from grids. Is there any tricks to keep sections stick on } the coated or bare nickel grids. } } Any suggestions are really appreciated! } } Shanling } } Shanling Shi } Advanced Imaging & Measurement } Unilever Research & Development -Edgewater } 45 River Road } Edgewater, NJ 07020 } 201-840-2340 } Shanling.Shi-at-unilever.com } } } }
Could you use OCT compound? The Oxford reps say it can be mixed with a graphite powder. I have never got round to trying it.
Dave
On Thu, 20 Mar 2003 11:03:04 +0000 "David.Mallard-at-bristol.ac.uk"-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 } -----------------------------------------------------------------------. } } } SEM -cryo. need info on conduction by Leit-C CCC at low temperatures } } I am looking at samples of ice (glacier & artificial chemical doped) } using SEM with cryo-stage. } } The samples are stuck to stubs using Leit-C conductive carbon cement. A } co-worker has suggested to me that carbon has greatly reduced } electrical conduction at liquid nitrogen temperatures but could not } remember where he had seen it. Does anyone have any information on this? } Thanks, } } David Mallard } Department Earth Sciences } Bristol University, UK }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
On Thu, 20 Mar 2003 11:03:04 +0000 "David.Mallard-at-bristol.ac.uk"-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 } -----------------------------------------------------------------------. } } } SEM -cryo. need info on conduction by Leit-C CCC at low temperatures } } I am looking at samples of ice (glacier & artificial chemical doped) } using SEM with cryo-stage. } } The samples are stuck to stubs using Leit-C conductive carbon cement. A } co-worker has suggested to me that carbon has greatly reduced } electrical conduction at liquid nitrogen temperatures but could not } remember where he had seen it. Does anyone have any information on this? } Thanks, } } David Mallard } Department Earth Sciences } Bristol University, UK }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
The 2003 Spring Meeting of THE TEXAS SOCIETY FOR MICROSCOPY “Embracing All Forms of Microscopy”
April 3, 4, 5, 2003
Meeting will be held in Hubbard Hall on the campus of Texas Woman’s University, Denton, TX.
THURSDAY WORKSHOP Microwave Techniques Presented by Rick Giberson, Research and Development Manager, Ted Pella, Inc. Workshop sponsored by Ted Pella, Inc. and TSM
MSA LAS-SPONSORED SPEAKER FRIDAY, APRIL 4, 2003 HUBBARD HALL TWU - DENTON CAMPUS Dr. Lucille A. Giannuzzi, Ph.D., Associate Professor, Mechanical, Materials & Aerospace Engineering and Director, UCF/Cirent Materials Characterization Facility, University of Central Florida, Orlando, FL. Topic: "Focused Ion Beam Specimen Preparation for Everything". Focused ion beam techniques have been developed
to prepare site-specific specimens in a reproducible and rapid manner for SEM and TEM analysis. The conventional and the lift-out methods of specimen preparation will be discussed and FIB applications on a wide range of materials will be presented. The usefulness for using the FIB for solving scientific research problems will be emphasized.
Registration forms, hotel information, and author's instructions can be found on our website, http://www.texasmicroscopy.org/ . Questions about the program can be directed to Jo Taylor, Program Chair, at jtaylor-at-sfasu.edu.
I have a group looking at very early germinated Arabidopsis sp. They are having problems with good infiltration of the Epon-Araldite epoxy. While they are out searching the literature for solutions, I thought I'd drop a note to the list. Any suggestions for an infiltration protocol for these tiny plants? Spurr's? Dice up those little seeds?
TIA
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
On Thursday, March 20, 2003, at 03:03 AM, "David.Mallard-at-bristol.ac.uk"-at-sparc5.microscopy.com wrote: } } SEM -cryo. need info on conduction by Leit-C CCC at low temperatures } } I am looking at samples of ice (glacier & artificial chemical doped) } using SEM with cryo-stage. } } The samples are stuck to stubs using Leit-C conductive carbon cement. A } co-worker has suggested to me that carbon has greatly reduced } electrical conduction at liquid nitrogen temperatures but could not } remember where he had seen it. Does anyone have any information on } this? } Thanks, } Dear David, I do not know the properties of the cement you mention, nor do I have at hand any quantitative data on the conductivities of carbon at 293 K vs 77 K; however, the conductivity of carbon evaporated films at 77 K is sufficient for TEM observations at that temp. The conductivity of carbon at 4.2 K is not adequate for observations, and maybe that is what your co-worker remembers. On the other hand, SEM and TEM are sufficiently different that carbon could also be inadequate for SEM observations at 77 K. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
After 24 years of managing our small facility as a collaborative effort with interested faculty, I now have a new supervisor with little technical expertise. Our facility consists of an older TEM, a new SEM, a new deconvolution scope, and a Lieca SP1 confocal. I don't want to air our dirty laundry but I need a little help in justifying our equipment to the new boss. Now, she is proud of the fact that she still uses PhotoShop 3.0 and states that the best way to get a good picture is to take a good picture and then don't mess with it. While there is certainly some truth in the second fact, it is also very naive. And the first fact classifies her as a Luddite. Her research centers around the confocal and the deconvolution scope. So the move is on to dump the TEM and give the Digital Darkroom to the department Network Admin. This will free me up to tackle confocal projects for her and spend more time dusting. Yes, after 24 years of managing the facility on my own my new boss has actually given me a dusting schedule. However, she is quick to point out that this is not micro-managing. Alright, I digress. I need to justify having a couple graphics workstations, a slide scanner, a flat bed scanner with transparency tray, and a photo-quality printer. She would give this equipment to the Network Admin who would integrate it with the common computer room. I treat this equipment like I do optical equipment. I am concerned that once relocated to a common room, with no supervision, it will rapidly deteriorate. I also take exception with the apparently wide-spread belief that the computer support staff is the best source of information on digital imaging. I guess because it is digital?
So how do I keep those pieces of equipment I deem necessary for the facility? I think we should have a photo-quality printer. I think we need the scanner with tray for those increasingly rare occasions that we need to scan in a TEM neg or an illustration for a PowerPoint slide or publication. I find that I am not good at making an argument for what seems blindingly obvious to me. I just sputter and fume and ask, "... how can anyone have an Imaging Facility without a couple workstations and printers?". Or a TEM.
And then there is always the possibility she is right. Maybe we don't need to do this stuff to the standards I embrace. Should I convert to her mantra: "Do more, less well."?
Any good arguments out there that have worked for any of you?
I have an ageing Jeol JSM840 SEM (~20 years old) with a rapidly fading viewing screen and with no framestore. I cannot get a replacement viewing screen so am looking at other options such as getting a third part image grabbing system that would allow tv rate viewing of a reasonable sized image on a pc screen for focussing etc. A framestore for averaging would also be useful. I don't need the system for capturing digital images, I can already do that via my EDS system, it's the viewing screen that is the problem. The EDS capture system does not allow tv rate viewing.
The Deben Pixie 3000 system appears to do what I want but I'm interested to know of any other similar systems available. I'd also like to hear comments from users of the Deben system.
Cheers
Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
David Carbon has very useful conductivity at 77K, though 100K would be more typical of an LTSEM system cooled by LN2. This is easy to confirm in the lab with a multimeter. My measurements indicate that a 3mm dia. graphite rod 70mm long has a resistance of 1.2 ohms at room temperature and 2.1 ohms at 77K. No dramatic conductivity loss, therefore.
Leit-C is solvent-based paint, and expensive too. Is it really the best choice for your purpose, I wonder. I use TissueTek or aqueous colloidal graphite (Aquadag) to attach ice samples to stubs. Effective and much cheaper. Best wishes Chris
On 20 Mar 03, at 15:00, Bill Tivol 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 } ---------------------------------------------------------------------- } -. } } } } On Thursday, March 20, 2003, at 03:03 AM, } "David.Mallard-at-bristol.ac.uk"-at-sparc5.microscopy.com wrote: } } } } SEM -cryo. need info on conduction by Leit-C CCC at low temperatures } } } } I am looking at samples of ice (glacier & artificial chemical doped) } } using SEM with cryo-stage. } } } } The samples are stuck to stubs using Leit-C conductive carbon } } cement. A co-worker has suggested to me that carbon has greatly } } reduced electrical conduction at liquid nitrogen temperatures but } } could not remember where he had seen it. Does anyone have any } } information on this? Thanks, } } } Dear David, } I do not know the properties of the cement you mention, nor do I have } } at hand any quantitative data on the conductivities of carbon at 293 K } vs 77 K; however, the conductivity of carbon evaporated films at 77 K } is sufficient for TEM observations at that temp. The conductivity of } carbon at 4.2 K is not adequate for observations, and maybe that is } what your co-worker remembers. On the other hand, SEM and TEM are } sufficiently different that carbon could also be inadequate for SEM } observations at 77 K. Yours, Bill Tivol EM Scientist and Manager } Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 } California Institute of Technology Pasadena CA 91125 (626) 395-8833 } tivol-at-caltech.edu } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 /8669 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
This is a rough one. We have just stated a new EM lab (less than two years operational) Our TEM (Which is new!) is not functioning due to the SIS camera being away for repairs under warranty now for some time. The EDS has been sent back twice now and I hope this time around the repairs will be successful. Being in Africa does have it pro's to like the weather, the birds and wildlife. We also have a lot more freedom in research. I am seeing forward to a prosperous year full of publications with EM input and well trained motivated users! (my whish list and also the units goal)
Thus if your TEM is running this is already a blessing. This alone (service contract cost depending) should be enough to motivate for keeping it. We are in a process to monitoring all publications on all instruments as a justification to keep it running or to replace it should the kneed arise.
My wife is brilliant at management and she helps me a lot. Her motto is always: "if you motivate something well enough you can cell ice to an Eskimo."
This is the most important point of all: Since she is now directing the facility, please go ask her to give the Objectives, Goals and mission statement for the facility to you in writing (Her view). If possible ask her also for a 5, and 10 Year plan for the facility. Before attempting to motivate anything think things over carefully. You two must be able to work in peace since strive will destroy the facility quickly and drive users away! With unity much can be achieved since her signature is required most likely on all purchase requests, motivations and budget expenditure.
When you do the motivation, link your instruments to the objectives and goals of your institute as well as the ones you got from her. Let her see that you are on her side and you would like to see these goals achieved.
If you can provide a list of publications where each instrument was involved in over its lifespan as a percentage of the total number of publications published in your institution it should help to explain the kneed of each instrument, scanner, SG's etc...
You can probably get a letter from the IT guys to state that your application, software, set-up and configuration is unique to the institution and it serve the best to keep it out the hands of ordinary villains to prevent expensive reconfiguration and repair. I had to fight to keep them out without driving them away since I kneed there help but not there software upgrades etc!
If you have a good relationship with your research office ask them for a opinion in writing.
It is worth it to do a survey in the institute to all relevant users, past users and possible users including head of departments. Things worth getting a answer on are questions like: What do you expect from the facility, What instruments would you like to be present in the unit, Past usage, usefulness of input in their research. Possible future utilisation of your unit. Suggestions to improve the service as well as suggestions wishes for instruments/capability's currently not present. If you can get postgads promised to instruments and research programs including your facility the better. I know that is not always possible but if you do not ask you will not know. We did that at a previous institute where I was employed. This was a very useful exercise. This helped to direct the unit and improve on our service. These documents are also a very useful source for equipment motivation when it is combined into a format top management can understand.
Hopefully from this information the wise path to follow will be much more clearer to all.
Good luck.
-----Original Message----- } From: Rick Harris [mailto:raharris-at-ucdavis.edu] Sent: Friday, March 21, 2003 1:08 AM To: microscopy-at-sparc5.microscopy.com
After 24 years of managing our small facility as a collaborative effort with interested faculty, I now have a new supervisor with little technical expertise. Our facility consists of an older TEM, a new SEM, a new deconvolution scope, and a Lieca SP1 confocal. I don't want to air our dirty laundry but I need a little help in justifying our equipment to the new boss. Now, she is proud of the fact that she still uses PhotoShop 3.0 and states that the best way to get a good picture is to take a good picture and then don't mess with it. While there is certainly some truth in the second fact, it is also very naive. And the first fact classifies her as a Luddite. Her research centers around the confocal and the deconvolution scope. So the move is on to dump the TEM and give the Digital Darkroom to the department Network Admin. This will free me up to tackle confocal projects for her and spend more time dusting. Yes, after 24 years of managing the facility on my own my new boss has actually given me a dusting schedule. However, she is quick to point out that this is not micro-managing. Alright, I digress. I need to justify having a couple graphics workstations, a slide scanner, a flat bed scanner with transparency tray, and a photo-quality printer. She would give this equipment to the Network Admin who would integrate it with the common computer room. I treat this equipment like I do optical equipment. I am concerned that once relocated to a common room, with no supervision, it will rapidly deteriorate. I also take exception with the apparently wide-spread belief that the computer support staff is the best source of information on digital imaging. I guess because it is digital?
So how do I keep those pieces of equipment I deem necessary for the facility? I think we should have a photo-quality printer. I think we need the scanner with tray for those increasingly rare occasions that we need to scan in a TEM neg or an illustration for a PowerPoint slide or publication. I find that I am not good at making an argument for what seems blindingly obvious to me. I just sputter and fume and ask, "... how can anyone have an Imaging Facility without a couple workstations and printers?". Or a TEM.
And then there is always the possibility she is right. Maybe we don't need to do this stuff to the standards I embrace. Should I convert to her mantra: "Do more, less well."?
Any good arguments out there that have worked for any of you?
Dear Rick, Definitely spurr's is the best. I do not how hard is the seed coat, You could try araldite-DDSA-DMP mixture. after fixation and dehydration, PO etc, Infilterate with 20% resin mix in PO and leave your sample vial open overnight. Next day(~20h) change it to fresh 100% resin mix for about 2hrs before embedding. curing, usual at 60 deg C for 72hrs. Shashi Singh Scientist CCMB Hyderabad-India
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I have a group looking at very early germinated Arabidopsis sp. They are having problems with good infiltration of the Epon-Araldite epoxy. While they are out searching the literature for solutions, I thought I'd drop a note to the list. Any suggestions for an infiltration protocol for these tiny plants? Spurr's? Dice up those little seeds?
TIA
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
__________________________________________________ Do you Yahoo!? Yahoo! Platinum - Watch CBS' NCAA March Madness, live on your desktop! http://platinum.yahoo.com
Peter No, I did a two-point measurement with the rod connected at both ends. The rod was immersed in liquid nitrogen while remaining connected, so the resistances are comparable. The figures I quote are the resistance of the specified length of the 3mm carbon rod, not the resistivity of carbon, which is usually quoted as ohms per square. Chris
} David; } } In doing this measurement of electrical resistivity, did you use a } "four point Kelvin" measurement to determine sheet resistance? It's } the only way I know of to know and compare resistances unless all } things in the comparison are held the same, i.e. thickness, width and } length. } } Peter Tomic } } } ----- Original Message ----- } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } To: {David.Mallard-at-bristol.ac.uk} ; "Bill Tivol" {tivol-at-caltech.edu} } Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, March 21, 2003 } 3:45 AM Subject: Re: SEM -cryo. need info on conduction by Leit-C CCC } at low temperatures } } } } -------------------------------------------------------------------- } } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } } ---. } } } } } } David } } Carbon has very useful conductivity at 77K, though 100K would be } } more typical of an LTSEM system cooled by LN2. This is easy to } } confirm in the lab with a multimeter. My measurements indicate that } } a 3mm dia. graphite rod 70mm long has a resistance of 1.2 ohms at } } room temperature and 2.1 ohms at 77K. No dramatic conductivity loss, } } therefore. } } } } Leit-C is solvent-based paint, and expensive too. Is it really the } } best choice for your purpose, I wonder. I use TissueTek or aqueous } } colloidal graphite (Aquadag) to attach ice samples to stubs. } } Effective and much cheaper. Best wishes Chris } } } } On 20 Mar 03, at 15:00, Bill Tivol 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 } } } ------------------------------------------------------------------ } } } ---- -. } } } } } } } } } } } } On Thursday, March 20, 2003, at 03:03 AM, } } } "David.Mallard-at-bristol.ac.uk"-at-sparc5.microscopy.com wrote: } } } } } } } } SEM -cryo. need info on conduction by Leit-C CCC at low } } } } temperatures } } } } } } } } I am looking at samples of ice (glacier & artificial chemical } } } } doped) using SEM with cryo-stage. } } } } } } } } The samples are stuck to stubs using Leit-C conductive carbon } } } } cement. A co-worker has suggested to me that carbon has greatly } } } } reduced electrical conduction at liquid nitrogen temperatures } } } } but could not remember where he had seen it. Does anyone have } } } } any information on this? Thanks, } } } } } } } Dear David, } } } I do not know the properties of the cement you mention, nor do I } } } have } } } } } } at hand any quantitative data on the conductivities of carbon at } } } 293 K vs 77 K; however, the conductivity of carbon evaporated } } } films at 77 K is sufficient for TEM observations at that temp. } } } The conductivity of carbon at 4.2 K is not adequate for } } } observations, and maybe that is what your co-worker remembers. On } } } the other hand, SEM and TEM are sufficiently different that carbon } } } could also be inadequate for SEM observations at 77 K. Yours, Bill } } } Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility } } } Broad Center, Mail Code 114-96 California Institute of Technology } } } Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu } } } } } } } } } } } } ========================================== } } Dr. Chris Jeffree } } University of Edinburgh } } BIOSEM - Biological Sciences Electron Microscope Facility } } Institute of Cell and Molecular Biology } } Waddington Building } } King's Buildings, Mayfield Road } } EDINBURGH, EH9 3JN, Scotland, UK } } Tel. #44 (0) 131 650 5554 /8669 } } FAX. #44 (0) 131 650 6563 } } Mobile 07710 585 401 } } email c.jeffree-at-ed.ac.uk } } ========================================= } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 /8669 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
More questions! How do you decide whether to use TissueTek or aqueous colloidal graphite (Aquadag) for an application? Where do get the latter?
Dave
On Fri, 21 Mar 2003 08:45:18 -0000 Chris Jeffree {cjeffree-at-srv0.bio.ed.ac.uk} 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 } -----------------------------------------------------------------------. } } } David } Carbon has very useful conductivity at 77K, though 100K would be } more typical of an LTSEM system cooled by LN2. This is easy to } confirm in the lab with a multimeter. My measurements indicate } that a 3mm dia. graphite rod 70mm long has a resistance of 1.2 } ohms at room temperature and 2.1 ohms at 77K. No dramatic } conductivity loss, therefore. } } Leit-C is solvent-based paint, and expensive too. Is it really the } best choice for your purpose, I wonder. I use TissueTek or aqueous } colloidal graphite (Aquadag) to attach ice samples to stubs. } Effective and much cheaper. } Best wishes } Chris } } On 20 Mar 03, at 15:00, Bill Tivol 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 } } ---------------------------------------------------------------------- } } -. } } } } } } } } On Thursday, March 20, 2003, at 03:03 AM, } } "David.Mallard-at-bristol.ac.uk"-at-sparc5.microscopy.com wrote: } } } } } } SEM -cryo. need info on conduction by Leit-C CCC at low temperatures } } } } } } I am looking at samples of ice (glacier & artificial chemical doped) } } } using SEM with cryo-stage. } } } } } } The samples are stuck to stubs using Leit-C conductive carbon } } } cement. A co-worker has suggested to me that carbon has greatly } } } reduced electrical conduction at liquid nitrogen temperatures but } } } could not remember where he had seen it. Does anyone have any } } } information on this? Thanks, } } } } } Dear David, } } I do not know the properties of the cement you mention, nor do I have } } } } at hand any quantitative data on the conductivities of carbon at 293 K } } vs 77 K; however, the conductivity of carbon evaporated films at 77 K } } is sufficient for TEM observations at that temp. The conductivity of } } carbon at 4.2 K is not adequate for observations, and maybe that is } } what your co-worker remembers. On the other hand, SEM and TEM are } } sufficiently different that carbon could also be inadequate for SEM } } observations at 77 K. Yours, Bill Tivol EM Scientist and Manager } } Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 } } California Institute of Technology Pasadena CA 91125 (626) 395-8833 } } tivol-at-caltech.edu } } } } } } } ========================================== } Dr. Chris Jeffree } University of Edinburgh } BIOSEM - Biological Sciences Electron Microscope Facility } Institute of Cell and Molecular Biology } Waddington Building } King's Buildings, Mayfield Road } EDINBURGH, EH9 3JN, Scotland, UK } Tel. #44 (0) 131 650 5554 /8669 } FAX. #44 (0) 131 650 6563 } Mobile 07710 585 401 } email c.jeffree-at-ed.ac.uk } ========================================= }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
If you just want to see the interior of the wood, it should be easy enough
to split 5mm cubes with a single edge razor blade, knife or wood chisel along the longitudinal axis. You have your choice of exposing surfaces with orientations that are radial, tangential or something in between. (If
those terms don't mean anything to you, take a look at the Soc. for Wood Sci. & Tech. at http://www.swst.org/teach/set2/struct1.html for an excellent introduction to wood structure and terminology).
If you need to expose a cross-section, I'd echo the LN2 idea with a couple
of suggestions. For my wood science MS research, I had to break numerous 5mm square by 50mm "matchsticks" with as brash (perpendicular to the long axis and not jagged) a failure as possible for an SEM study. What worked best for me was to lightly score the wood on the tension side where I wanted the break and insert it into a small jig that clamped the matchstick on both sides of the score line. I immersed the jig w/ wood in LN2 until the boiling slowed significantly (may take a while, wood is a good insulator) and broke the sample immediately and energetically on removal from the LN2. If your pieces are only 5mm cubes, you may not be able to use such a jig, but perhaps you don't need the brash failure I did. Another factor I found important was the moisture content of the wood
- too low was a problem. No matter what technique you use, you'll likely encounter the variability of wood. Count on lots of replications!
Pete Eastman Microscopy Rohm and Haas Company 215 619-5229
"Tindall, Randy D." {TindallR-at-missouri.edu} 03/19/2003 09:13 AM
To: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} cc: {microscopy-at-sparc5.microscopy.com} bcc: Subject: RE: CPD or HMDS for wood blocks?
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Jonathan,
We recently looked at wood blocks in our FESEM and the only preparation we did was to freeze them in liquid nitrogen, then fracture them with pliars to expose the internal structure, followed by several days in a vacuum chamber to get as much moisture and air out as possible. We glued them down with copius amounts of conductive paint, then sputtered the bejesus out of them.
The structure was well preserved and we did find bacteria.
The only problem was breaking the harder pieces of wood. Some of the wood was ancient and very brittle, but the new pieces were tough.
Hope this is applicable to your problem.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Jon Krupp [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Tuesday, March 18, 2003 3:03 PM To: Microscopy-at-sparc5.microscopy.com
Hi again:
I have a guy who wants to look at the interior of some wood blocks using our conventional SEM. He is trying to see bacteria and anything else that might be hiding in there.
His blocks are pretty small, about 5mm cubes. I thought about CPD, but had second thoughts about how long to soak in CO2 etc. He did some experiments and it took a couple of hours for some dye to make it all the way into the middle of the block. Plus he has about 30 of these things to look at. That could add up to a lot of time and CO2.
I got an idea that HMDS might work for this project, but have no experience with it. I think that being able to soak the blocks for a long time in HMDS would be good, as would the chance to let them sublime to dryness over a long time without too much attention.
Do you think I am on the right track or is there something I am missing.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
On Thursday, March 20, 2003, at 07:12 PM, Dave Phelan wrote:
} I have an ageing Jeol JSM840 SEM (~20 years old) with a rapidly fading } viewing screen and with no framestore. I cannot get a replacement } viewing } screen so am looking at other options such as getting a third part } image } grabbing system that would allow tv rate viewing of a reasonable sized } image on a pc screen for focussing etc. A framestore for averaging } would } also be useful. I don't need the system for capturing digital images, } I can } already do that via my EDS system, it's the viewing screen that is the } problem. The EDS capture system does not allow tv rate viewing. } Dear Dave, If you have old screens or screen blanks, they can be recoated with phosphor. There are companies that do this; I don't have access to their names or addresses, but I am pretty sure I have posted at least one to the list in the past. You can also recoat the screen blank yourself, and I think I posted a procedure to the list also. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Dave Phelan wrote: ============================================================================ ======== I have an ageing Jeol JSM840 SEM (~20 years old) with a rapidly fading viewing screen and with no framestore. I cannot get a replacement viewing screen so am looking at other options such as getting a third part image grabbing system that would allow tv rate viewing of a reasonable sized image on a pc screen for focussing etc. A framestore for averaging would also be useful. I don't need the system for capturing digital images, I can already do that via my EDS system, it's the viewing screen that is the problem. The EDS capture system does not allow tv rate viewing.
The Deben Pixie 3000 system appears to do what I want but I'm interested to know of any other similar systems available. I'd also like to hear comments from users of the Deben system. ============================================================================ ========= Another alternative is the passive image capture software called Spectrum Mono, as described on URL http://www.2spi.com/catalog/software/spectrum.shtml
It does what you want it to do, and, it is perhaps the lowest cost alternative for a product of this type. On most older SEMs it is easily installed, either by the user or the preferred service engineer. We have had the software running on one of our own Model 840 JEOL SEMs and it seems to perform up to expectations.
Disclaimer: SPI Supplies has distributed this "passive" image capture software for some time and we have a vested interest in its promotion.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
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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Can anyone recommend a source of 5-10L LN2 portable dewars? These are for filling a 10L EDS detector. The LN2 will be transported by car.
Before I introduce myself, I wish to inform you that this letter is not a hoax mail and I urge you to treat it serious. I am Dr. Muhammad Khalil Hasan, a banker with the Mashreq Bank Plc Dubai, UAE. I am the Accounts officer late Mr. Robert Chapman the African Area Director of SIL International, who unfortunately died in the crash of Kenya Airways Flight 431 in Abidjan, Ivory Coast, January 30 2000. You will read more stories about the crash on visiting this website, news.airwise.com/airlines/archive/2000/kenya2000.html and also in this website, where Chapman's company talked about his death in the Kenya crash. You shall as well find the pictures of Chapman and his wife there. Mr. Chapman was from Hamilton, Ontario Canada.
Since the death of Chapman, I as his accounts officer in the bank, have made several enquiries to locate his only surviving relation, without any success. I came across your name and contact, on the course of my personal searching for Chapman's relations, so I decided to contact you for this project.
I am contacting you to assist in repatriating and securing the wealth left behind in a fixed deposit account by Robert, before they get confiscated or declared unserviceable by my bank. The board of my bank, has issued a notice that after 2 months from now and no next of kin shown up for the claim, the funds will be confiscated and declared unserviceable. Since I and my team have been unsuccessful in locating Robert's relatives for sometime now, I seek your consent to present you as the Next of Kin of the deceased since you are a foreigner, so that the proceed of this deposit valued at $USD7.5 Million Dollars can be released to you.
The bank will release the funds to any foreigner who has all related information/documents to the bank account. I am in charge of this matter in my bank, because I am his accounts officer. Your application will be directed to my department for verification and approval. Everything is under my control.
I shall provide you all the information and copy of the certificate of deposit issued to Robert when he deposited the funds. I shall also involve a good attorney who shall represent you in all the appropriate offices for the claim. Please, find in the attachment my work ID Card. This is just to proof to you that my proposal is genuine. I also have all necessary information we need for the claim and once the money is transferred to you, I shall destroy all the documents used for the claim and leave no traces. After everything, you shall have 40% of the total sum,while 60% for me.
All requires is your honest cooperation to enable us see this business successful. I guarantee that this will be executed under a legitimate arrangement that will protect you from any breach of the law. Robert was a very good man and it is not wise to allow his hard earned wealth to be stolen by the greedy directors of my bank.
Further details awaits your response by email. PLEASE, TREAT THIS PROPOSAL AS TOP SECRET. Please, confirm the receipt of my Id Card in the attachment, as it was difficult to attach.
Tell her NO! Make no consigns not afoot to compromise or be her friend just tell her no.
When you end up in her supervisors offices explaining the dusting duties, her ludilite ways and designs to enslave you. If he tells you to go along to get along say no. Let her find some other domestic servant to do her work. and call a head hunter.
Unless dusting is what you should be doing I would think if you are any good you can find a better paying job down the hall and let here dust her own lab.
The only thing is you have to mean no you can say it and not mean it shows.
One of the reasons women get less pay is they will put up with a lot more abuse on the job than most a,m/
No is an extremely powerful word that is not used near enough. If the whole lab will tell her no se will be gone by the end of the moth unless she is sleeping wiht the boss. Then it will take another month or two. In the mean time a match stem propping the valve on a tire open will annoy here. do it to two tires cost here 45 minutes that day.A 10 dollar bill will get the doomster sat where her car can't get over night. A 40 dollar tip will get a pitcher of ice water down the front of her dress as she gets up to speak at a meeting..
Even the densest manager gets the clue some where along the line.
Bad management is on of the most embedded instructions in our country and to letters NO can bring them to their knees if everyone cooperates and be rid of them.
They may be a great tech and like the work give them power over other can turn Dr. Jeckel into Mr. Hyde.
Being a bad manger is only a third the mangers fault. A third or more rests with their boss and third rests with the people that work for them. You were looking for a job when you found this one and the next one should pay better with new challenges and new manure piles to afford.
good luck
Gordon Couger Stillwater, OK www.couger.com/gcouger ----- Original Message ----- } From: "Rick Harris" {raharris-at-ucdavis.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 20, 2003 5:07 PM
I am in need of a scoring wheel cartridge for a Leica Reichert Knifemaker II. Please reply with source contact info.
Thanks.
William C. Hamlett, Ph.D. Professor of Anatomy & Cell Biology Indiana University School of Medicine Notre Dame, Indiana 46556, USA e-mail whamlett-at-nd.edu Telephone 574-631-7194 FAX 574-631-7821 http://www.nd.edu/~whamlett
This is to inform you that the Call for Abstracts for the forthcoming International Meeting on Applied Physics (APHYS-2003), to be held during October 14-18th 2003 in Badajoz (Spain), is open. All the information regarding this interdisciplinary conference can be found at the Conference website
www.formatex.org/aphys2003/aphys2003.htm
Some of the topics to be covered will be
- Surfaces, Interfaces and Colloids - Imaging Techniques, Microscopy - Nano-sciences and Technologies - Biophysics and Biophysical Chemistry - Materials Science & Engineering, Applied Solid State Physics/Chemistry - Advanced and Fuctional Materials - Biomedical Engineering and Biomaterials - Biological & Medical Physics - Computational Physics - Radiation Physics, Applied Nuclear Physics/chemistry, Radiation Protection
In addition to the regular Scientific Program, several International Workshops will be held as pre-conference events. The following Workshops are presently confirmed
1. Workshop on Modern Applied Microscopy in Molecular and Cell Biophysics Research 2. International Interdisciplinar Workshop on Bioengineered Non-crystalline Solids 3. Workshop on Interfaces in Colloidal and Particulate Systems 4. International Workshop on Radiation Protection and Dosimetry
The Conference will be specifically interested in receiving reports on Interdisciplinary researches relating Physics with other Sciences such as Biology, Chemistry, Information Technology, Medicine, etc or relating different Physics areas. In other words, we are specially (but not exclusivelly) interested in reports applying the techniques, the training, and the culture of physics to research areas usually associated with other scientific and engineering disciplines
APHYS-2003 will also serve as a platform to search for partners for transnational collaboration projects, specially for the EU Sixth Framework Program (NETworks of Excellence and Integrated Projects). "Projects Presentations" and "Call for Partners" presentations proposals are therefore encouraged and welcomed. If you are interested in taking part of this Conference feature, please send us the corresponding form available at the website.
In addition to the "traditional" oral contributed and posters presentation, a Virtual Participation modality has been established for those researchers unable to attend it in person. A limited number of works can be presented in this way. Please refer to the Conference website for details.
If you are interested in taking part of APHYS-2003, please send us your PRE-REGISTRATION FORM (at the main website of the conference) as soon as possible. The pre-registration form is also available through the direct URL http://www.formatex.org/aphys2003/preregistration.htm
Deadline for abstracts submission is April 30th 2003 (for oral presentations proposals; for posters additional time will be provided) although we highly recommend you to submit your abstract as soon as possible to avoid saturation during the days before the deadline.
Proceedings Accepted and presented papers will be reviewed for publication in special issues of several international Journals: - Journal of Microscopy - Journal of Non-crystalline Solids - Applied Surface Science - Colloids and Surfaces A - Powder Technology (to be confirmed) - Microelectronics Journal - Physica Scripta - Radiation Protection Dosimetry - Applied Physics A (Materials Science & Processing, to be confirmed). - Biomedial Materials and Engineering
Also a book "Advances in Applied Physics" will be published by an international publisher (Kluwer or the American Institute of Physics, with those papers accepted for presentation but not suitable for the journal issues.For up-to-date information on publications participating at the Conference as publishers, please visit regularly the Conference website (Proceedings sections).
For any question or suggestion, please do not hesitate to contact us at secretariat-at-formatex.org, or visit www.formatex.org/aphys2003/aphys2003.htm (Bookmark the page!!) We would also appreciate if could disseminate this Call for Papers through your Department or Institution.
We hope to meet you at this exciting and interdisciplinar international meeting!
SUV would be fine versus car. I thought that there were Dewars that could be filled and had pressure relief vents. Then, comes the problem of getting the LN2 out of the Dewar.
This is a tough area to tackle for a low use system. I suspect EDS would be used a half a day per week, perhaps less. Total of around 100 hours per year. For the non-use periods, the detector would be empty. This is suppose to be OK.
gary g.
At 09:42 AM 3/22/2003, you wrote: } Gary } There is really only one way to carry liquid nitrogen safely by car } and that is outside it. A pick-up truck, or SUV with a rear transport } cage for a dewar would be ideal, but if liquid nitrogen spills inside } the car you, your passengers and other road users are in mortal } danger. Taylor-Wharton manufacture dry shippers that can safely } transported inside cars and aircraft, but unfortunately they are } designed as refrigerators, and will not release liquid nitrogen when } inverted. They are therefore useless for refilling your EDS } detector. } Chris } } Dr. Chris Jeffree } Inveresk Cottage } 26, Carberry Road } Inveresk } Musselburgh } Midlothian } EH21 8PR } Tel: +44 131 665 6062 } FAX +44 131 653 6248 } Mobile 07710 585 401 } ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} } To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} } Sent: Saturday, March 22, 2003 2:20 AM } Subject: LN2 dewar } } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 source of 5-10L } } LN2 portable dewars? These are for filling } } a 10L EDS detector. The LN2 will be } } transported by car. } } } } Any ideas? } } } } tnx, } } gary g. } } } }
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Gary Gaugler wrote: ============================================================================ SUV would be fine versus car. I thought that there were Dewars that could be filled and had pressure relief vents. Then, comes the problem of getting the LN2 out of the Dewar.
This is a tough area to tackle for a low use system. I suspect EDS would be used a half a day per week, perhaps less. Total of around 100 hours per year. For the non-use periods, the detector would be empty. This is suppose to be OK. ============================================================================ == I thought that in the USA at least, since liquid nitrogen is considered to be a "dangerous goods" material, in order to transport it, one would need to be a licensed "dangerous goods" transporter. For example those who deliver liquid nitrogen have their trucks placarded with the green flammable gas placard. This is all highly regulated by the US DOT.
Is it not similar to the case for transporting hazardous waste, one needs to be licensed to carry it in a vehicle?
I am not the last word on this, however, and there could be some exemptions, perhaps for personal use, but if there are, I have not heard of any. We all have to make our own decisions about such things, but one should be sure that laws and/or regulations are not being violated. In the event of an accident, enormous legal liability for someone could be the result (not a legal opinion, just a lay presumption).
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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by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id LAA20580 for dist-Microscopy; Sun, 23 Mar 2003 11:50:19 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id LAA20576 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Sun, 23 Mar 2003 11:49:48 -0600 (CST) Received: from priv-edtnes04.telusplanet.net (outbound02.telus.net [199.185.220.221]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id LAA20569 for {Microscopy-at-sparc5.microscopy.com} ; Sun, 23 Mar 2003 11:49:36 -0600 (CST) Received: from interchange.ubc.ca ([142.179.32.64]) by priv-edtnes04.telusplanet.net (InterMail vM.5.01.05.17 201-253-122-126-117-20021021) with ESMTP id {20030323174356.LDUN8012.priv-edtnes04.telusplanet.net-at-interchange.ubc.ca} ; Sun, 23 Mar 2003 10:43:56 -0700
Rick, There are a number of factors to consider. It would help to know what parts are not infiltrated well to determine the cause of the problems.
Spurr's is definitely easier to get into plant material. If the seeds are still intact it will be hard to get any resin in. You stated that they are early germinants. Does this mean that they have broken through the seed coat? If they haven't, you may have to abrade the seeds in some way to get fixation and infiltration. Larger seeds can be shaken over sandpaper, but this may not work with the smaller seeds. (Dicing the seeds may work but could smash the cells.) Consider extending the infiltration time to several days to a week. I have had to infiltrate for more than a week to get successful embedding of some woody material. You will have to make regular changes of resin, but it does make a difference.
I don't know your protocol, but another factor may be fixation and especially dehydration. You have to use extended fixation and dehydration times to allow hydrophobic resin to penetrate plant material. Often the woody parts retain water and the resin won't penetrate.
If you have access to a lab microwave then you should consider trying infiltration in there. It really helps!
Good luck,
Kim {} {} {} {} {} Kim Rensing PhD Department of Botany, UBC 6270 University Blvd. Vancouver BC, Canada V6T 1Z4
On Thursday, March 20, 2003, at 02:02 PM, Rick Harris wrote: } } I have a group looking at very early germinated Arabidopsis sp. They } are having problems with good infiltration of the Epon-Araldite epoxy. } While they are out searching the literature for solutions, I thought } I'd drop a note to the list. Any suggestions for an infiltration } protocol for these tiny plants? Spurr's? Dice up those little seeds? } } TIA } } } Rick A. Harris, Director } Microscopy and Imaging Facility
Rick, There are a number of factors to consider. It would help to know what parts are not infiltrated well to determine the cause of the problems.
Spurr's is definitely easier to get into plant material. If the seeds are still intact it will be hard to get any resin in. You stated that they are early germinants. Does this mean that they have broken through the seed coat? If they haven't, you may have to abrade the seeds in some way to get fixation and infiltration. Larger seeds can be shaken over sandpaper, but this may not work with the smaller seeds. (Dicing the seeds may work but could smash the cells.) Consider extending the infiltration time to several days to a week. I have had to infiltrate for more than a week to get successful embedding of some woody material. You will have to make regular changes of resin, but it does make a difference.
I don't know your protocol, but another factor may be fixation and especially dehydration. You have to use extended fixation and dehydration times to allow hydrophobic resin to penetrate plant material. Often the woody parts retain water and the resin won't penetrate.
If you have access to a lab microwave then you should consider trying infiltration in there. It really helps!
Good luck,
Kim {} {} {} {} {} Kim Rensing PhD Department of Botany, UBC 6270 University Blvd. Vancouver BC, Canada V6T 1Z4
On Thursday, March 20, 2003, at 02:02 PM, Rick Harris wrote: } } I have a group looking at very early germinated Arabidopsis sp. They } are having problems with good infiltration of the Epon-Araldite epoxy. } While they are out searching the literature for solutions, I thought } I'd drop a note to the list. Any suggestions for an infiltration } protocol for these tiny plants? Spurr's? Dice up those little seeds? } } TIA } } } Rick A. Harris, Director } Microscopy and Imaging Facility
JEOL will have your picture tube (CRT) for a replacement. If they don't by some miracle, then contact Richardson Electronics at www.rell.com or (630)208-2200. Use information from your SEM CRT label (monitor pulls out, label is on the side of a CRT). You can get new CRT form Richardson for couple-three hundred $. Make sure they are aware that CRT phosphor must have longer decay time (phosphor type is also stated on the CRT label). Longer phosphor decay time is not critical, but helpful.
If you in the mood, get a monochrome video monitor (such as security monitor), and connect TV video from SEM to it.
Countless video frame grabbers with frame averaging function are available- too many to list. What features and what price range are you looking for?
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons.
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----- Original Message ----- } From: "Dave Phelan" {emudp-at-mail.newcastle.edu.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 20, 2003 10:12 PM
Hi, Bill
Could you re-post the directions for a home-cooking procedure? I didn't think it was possible to recoat CRTs.
cheers
rtch
} } } On Thursday, March 20, 2003, at 07:12 PM, Dave Phelan wrote: } } } I have an ageing Jeol JSM840 SEM (~20 years old) with a rapidly } } fading viewing screen and with no framestore. I cannot get a } } replacement viewing screen so am looking at other options such as } } getting a third part image grabbing system that would allow tv rate } } viewing of a reasonable sized image on a pc screen for focussing } } etc. A framestore for averaging would also be useful. I don't need } } the system for capturing digital images, I can already do that via } } my EDS system, it's the viewing screen that is the problem. The EDS } } capture system does not allow tv rate viewing. } } } Dear Dave, } If you have old screens or screen blanks, they can be recoated with } phosphor. There are companies that do this; I don't have access to } their names or addresses, but I am pretty sure I have posted at least } one to the list in the past. You can also recoat the screen blank } yourself, and I think I posted a procedure to the list also. Good } luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron } Microscopy Facility Broad Center, Mail Code 114-96 California } Institute of Technology Pasadena CA 91125 (626) 395-8833 } tivol-at-caltech.edu } }
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
} From: "Garber, Charles A." {cgarber-at-2spi.com} : : Gary Gaugler wrote: : ============================================================================ : SUV would be fine versus car. I thought that there were Dewars that could : be filled and had pressure relief vents. Then, comes the problem of getting : the LN2 out of the Dewar. : : This is a tough area to tackle for a low use system. I suspect EDS would be : used a half a day per week, perhaps less. Total of around 100 hours per : year. For the non-use periods, the detector would be empty. This is : suppose to be OK. : ============================================================================ : == : I thought that in the USA at least, since liquid nitrogen is considered to : be a "dangerous goods" material, in order to transport it, one would need to : be a licensed "dangerous goods" transporter. For example those who deliver : liquid nitrogen have their trucks placarded with the green flammable gas : placard. This is all highly regulated by the US DOT. : : Is it not similar to the case for transporting hazardous waste, one needs to : be licensed to carry it in a vehicle? : : I am not the last word on this, however, and there could be some exemptions, : perhaps for personal use, but if there are, I have not heard of any. We : all have to make our own decisions about such things, but one should be sure : that laws and/or regulations are not being violated. In the event of an : accident, enormous legal liability for someone could be the result (not a : legal opinion, just a lay presumption). : : Chuck : Liquid nitrogen should be much less dangerous the liquid oxygen and I see trucks that haul live fish with LOX tanks on them. They may be hauling the LOX without the proper permits. The people in the bait fish business are not particular careful about obeying the law. Artificial insemination technicians carry LN2 dewers all over the country as well.
To pump the LN2 put a pipe that reaches the bottom of the tank. Just close the vents and put a little current through a heater submerged in the nitrogen and the pressure will pump the LN2 out the pipe. Four psi should pump about 10 gallons a minutes or more though a 3/4 inch pipe to a container the same height as the LN2 tank.
If the temperature of boiling propane is low enough you could vaporize propane and dispose ot it in a flame and greatly reduce your storage and procurement problems. A propane tank has to be in sealed compartment in vehicle or mounted out it and the vehicle should not be parked in a building. It can legally be installed in vehicles.
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (spohnheimer-at-dalsemi.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, March 22, 2003 at 19:57:07 ---------------------------------------------------------------------------
Email: spohnheimer-at-dalsemi.com Name: John Spohnheimer
Organization: Dallas Semiconductor (Commercial)
Education: Graduate College
Location: Dallas, TX
Question: I am an engineer/physicist working with semiconductors & have used high-quality optical/SEM/TEM microscopes for ~25 years. Recently, a friend (also a physicist) asked why we can see sub-micron geometries (sometimes even one-two tenth micron diameter particles are visible) using optical microscopes with wavelengths nominally in the 1/2 micron range.
I argued that what we're really seeing is simply a phase interference pattern generated by the sub-micron particle, not the real particle itself. That's why most sub-resolution particles appear spherical in an optical microscope and often-times look completely different under SEM examination.
I'm curious if a professional in this field has a better/more accurate explanation.
I have learned your name from the Internet. We are the manufacturer&exporter in China, specializing in producing Bathroom Products. Please visit our website www.china-bathroom.com for more information. OEM is available. Please let me know if I may be of further assistance. ( E-mail: jimmy-at-china-genesis.com )
Yours sincerely, Mr. Jimmy Sky
Ningbo Genesis Industry Co., Ltd. Tel: 86-574-87330333 Fax: 86-574-87725245 Location: Ningbo city, Zhejiang province, China ( a port city, 150km from Shanghai )
If you think this is spam, please send email Subject "remove" to jimmy197809-at-hotmail.com.
I have learned your name from the Internet. We are the manufacturer&exporter in China, specializing in producing Bathroom Products. Please visit our website www.china-bathroom.com for more information. OEM is available. Please let me know if I may be of further assistance. ( E-mail: jimmy-at-china-genesis.com )
Yours sincerely, Mr. Jimmy Sky
Ningbo Genesis Industry Co., Ltd. Tel: 86-574-87330333 Fax: 86-574-87725245 Location: Ningbo city, Zhejiang province, China ( a port city, 150km from Shanghai )
If you think this is spam, please send email Subject "remove" to jimmy197809-at-hotmail.com.
To upgrade our old microscope (a double condenser lens VG HB501) we are looking for a second-hand parallel electron energy loss spectrometer (PEELS) or a second-hand Gatan Image Filter (GIF). Please e-mail any information to me if you have one. Thanks marie
----------------------------------------------------------------- Marie-Claude Cheynet Groupe Physique du métal LTPCM/ENSEEG/CNRS(UMR-5614) Domaine Universitaire BP75 Saint-Martin d'Hères 38402 e-mail mcheynet-at-ltpcm.inpg.fr tel : 33 (0)4.76.82.66.14 http://www.inpg.fr/LTPCM/ -------------------------------------------------------------------------------- -----------------
Greetings, This reflects a common misconception about the quantitative definition of 'resolution' in microscopy. For light microscopy, one is quite used to hearing that the 'resolution limit' is about 0.25 microns, or thereabouts. So it is natural to wonder how come it is pretty easy to see things like microtubules, which have a diameter around 1/10th of that limit. The answer lies in the definitition of resolution: that limit defines how close two objects can approach and still be resolved as two objects. When they get closer than the limit, then they are seen as one single object. However, the definition says nothing about the smallest object that can give rise to an image. That is limited only by the sensitivity of the detector. So objects like microtubules many times smaller than the resolution limit can be detected. If you measure their diameter, you will find that they measure up to whatever the diffraction limit is in your particular scope. When two of them happen to be closer than that limit, then they are detected as one object.
Hope this helps, Tobias } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (spohnheimer-at-dalsemi.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, } March 22, 2003 at 19:57:07 } --------------------------------------------------------------------------- } } Email: spohnheimer-at-dalsemi.com } Name: John Spohnheimer } } Organization: Dallas Semiconductor (Commercial) } } Education: Graduate College } } Location: Dallas, TX } } Question: I am an engineer/physicist working with semiconductors & } have used high-quality optical/SEM/TEM microscopes for ~25 years. } Recently, a friend (also a physicist) asked why we can see } sub-micron geometries (sometimes even one-two tenth micron diameter } particles are visible) using optical microscopes with wavelengths } nominally in the 1/2 micron range. } } I argued that what we're really seeing is simply a phase } interference pattern generated by the sub-micron particle, not the } real particle itself. That's why most sub-resolution particles } appear spherical in an optical microscope and often-times look } completely different under SEM examination. } } I'm curious if a professional in this field has a better/more } accurate explanation. } } Thank you. } } } ---------------------------------------------------------------------------
I looked around on the US DOT web site, and was a little surprised to find that it seems there is not, in the United States at least, a class of hazardous material called "Cryogen". The most useful document I found in my (brief) search was http://hazmat.dot.gov/files/registration/0203/regbrch2002.pdf which clearly lists the classes of materials that require placarding of vehicles and registration of carriers. My lay reading of this leads me to the conclusion that liq. N2 would be placarded as "Non-flammable gas" which is only regulated in quantities over 1000 pounds. It may, naturally, be different in other countries. Nevertheless, common sense must prevail, and that tells us that one must be thoughtful about transporting any quantity of the material. There will be other applicable safety regulations that apply to liquid nitrogen at all times, whether during transportation or not, (for example, occupational safety regulations), which I have not even begun to address here.
Once you add the "W" word to any material, though, the issue is **VERY** different. Even a microgram of waste is highly regulated (or so it seems!).
Tony.
At 04:07 PM 3/23/2003 -0600, Gordon Couger wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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When I was working in a lab with low-overhead - basically owned by someone with shallow pockets - I successfully transported LN2 in my car using a styrofoam cooler. I wasn't too keen on it and I don't recommend doing it, but I thought this may be worth mentioning. LN2 thermoses are somewhat expensive... I believe $200-$500, but worth it for a lab committed to doing EDS work.
If you must transport by car, keep the windows open or blower fan on if it's in the passenger cabin. I doubt that there are DOT issues involved. LN2 is not a substance one would consider highly hazardous, plus I believe DOT has jurisdiction only for amounts above a certain minimum. Only practical issues apply.
On the matter of occasional EDS use, one can simply fill LN2 when needed, allowing for proper detector cooldown (roughly 2-4 hours for my particular model). This method has to weighed against the thermal cycling that occurs, which is not good and potentially may - with time - damage the detector by delamination.
Stu Smalinskas Metallurgist SKF NATC Plymouth, Michigan
Gordon cougar wrote:
Liquid nitrogen should be much less dangerous the liquid oxygen and I see trucks that haul live fish with LOX tanks on them. They may be hauling the LOX without the proper permits. The people in the bait fish business are not particular careful about obeying the law. Artificial insemination technicians carry LN2 dewers all over the country as well.
To pump the LN2 put a pipe that reaches the bottom of the tank. Just close the vents and put a little current through a heater submerged in the nitrogen and the pressure will pump the LN2 out the pipe. Four psi should pump about 10 gallons a minutes or more though a 3/4 inch pipe to a container the same height as the LN2 tank.
If the temperature of boiling propane is low enough you could vaporize propane and dispose ot it in a flame and greatly reduce your storage and procurement problems. A propane tank has to be in sealed compartment in vehicle or mounted out it and the vehicle should not be parked in a building. It can legally be installed in vehicles.
Gordon Couger gcouger-at-couger.com
Dr. Chris Jeffree wrote:
} There is really only one way to carry liquid nitrogen safely by car } and that is outside it. A pick-up truck, or SUV with a rear transport } cage for a dewar would be ideal, but if liquid nitrogen spills inside } the car you, your passengers and other road users are in mortal } danger. Taylor-Wharton manufacture dry shippers that can safely } transported inside cars and aircraft, but unfortunately they are } designed as refrigerators, and will not release liquid nitrogen when } inverted. They are therefore useless for refilling your EDS } detector. } Chris } } Dr. Chris Jeffree } Inveresk Cottage } 26, Carberry Road } Inveresk } Musselburgh } Midlothian } EH21 8PR } Tel: +44 131 665 6062 } FAX +44 131 653 6248 } Mobile 07710 585 401
Gary Gaugler wrote:
} } Can anyone recommend a source of 5-10L } } LN2 portable dewars? These are for filling } } a 10L EDS detector. The LN2 will be } } transported by car. } } } } Any ideas? } } } } tnx, } } gary g.
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There's a long answer involving such great party conversation pieces as Rayleigh's criterion and Ernst Abbe's formula for resolution in any optical system by diffraction which is R=0.61 lambda/ n sine alpha.
The important bit in this context is n sine alpha. Where n is refractive index of the medium through which light/electrons pass from the specimen into the lens and sine alpha is the sine of the semi angle of the objective lens This combined figure is generally called the Numeric Aperture (NA) of the lens and even our old teaching microscopes have an NA of 1.28. In other words to make resolution smaller or better you need a high refractive index (eg immersion oil 1.4 or greater) and a big lens as close as possible (eg an oil immersion lens) which give a NA as large as possible.
Some quick maths tells you that a basic light microscope is capable of a resolution of 0.61/1.28 lambda which is less than half the wavelength of light. But in the e.m. with an aperture of 100um diam and focal length of ~3mm you are lucky to get a NA of better than about 0.01. Why the e.m. has such a poor numerical aperture is because of lens aberrations especially spherical which are only corrected by using a small central objective lens aperture.
Basically light microscope lens design is so good that resolution is only limited by diffraction effects whereas electromagnetic lenses are a compromise.
My apologies if I've oversimplified.
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: spohnheimer-at-dalsemi.com (by way of MicroscopyListServer)
Thanks to many who replied to my post about LN2 Dewars and transporting them. It is possible to do this. however, several have pointed out the issues surrounding temperature cycling of the EDS Dewar. The makers claim that this is not a problem--even with Si(Li) detectors. And the systems will power down the amplifier if LN2 is low or out. And, they will not allow the unit to power up if LN2 is not correct. But there seems to be issues surrounding the window. The makers claim that this is not so.
OK. Plan B. So what about the closed loop refrigerated EDS units like EDAX makes? Does anyone have any direct experience with these? Good or bad. I need to detect down to N.
I had heard that these Kleemenko cycle cryocoolers are notoriously unreliable. The pumps fail frequently. Perhaps also, there are sensitivity issues and low Z detection limits. I had looked at closed loop WDS systems before and got the same reliability feedback from users. How about EDAX EDS units?
This is a rough one. We have just stated a new EM lab (less than two years operational) Our TEM (Which is new!) is not functioning due to the SIS camera being away for repairs under warranty now for some time. The EDS has been sent back twice now and I hope this time around the repairs will be successful. Being in Africa does have it pro's to like the weather, the birds and wildlife. We also have a lot more freedom in research. I am seeing forward to a prosperous year full of publications with EM input and well trained motivated users! (my whish list and also the units goal)
Thus if your TEM is running this is already a blessing. This alone (service contract cost depending) should be enough to motivate for keeping it. We are in a process to monitoring all publications on all instruments as a justification to keep it running or to replace it should the kneed arise.
My wife is brilliant at management and she helps me a lot. Her motto is always: "if you motivate something well enough you can cell ice to an Eskimo."
This is the most important point of all: Since she is now directing the facility, please go ask her to give the Objectives, Goals and mission statement for the facility to you in writing (Her view). If possible ask her also for a 5, and 10 Year plan for the facility. Before attempting to motivate anything think things over carefully. You two must be able to work in peace since strive will destroy the facility quickly and drive users away! With unity much can be achieved since her signature is required most likely on all purchase requests, motivations and budget expenditure.
When you do the motivation, link your instruments to the objectives and goals of your institute as well as the ones you got from her. Let her see that you are on her side and you would like to see these goals achieved.
If you can provide a list of publications where each instrument was involved in over its lifespan as a percentage of the total number of publications published in your institution it should help to explain the kneed of each instrument, scanner, SG's etc...
You can probably get a letter from the IT guys to state that your application, software, set-up and configuration is unique to the institution and it serve the best to keep it out the hands of ordinary villains to prevent expensive reconfiguration and repair. I had to fight to keep them out without driving them away since I kneed there help but not there software upgrades etc!
If you have a good relationship with your research office ask them for a opinion in writing.
It is worth it to do a survey in the institute to all relevant users, past users and possible users including head of departments. Things worth getting a answer on are questions like: What do you expect from the facility, What instruments would you like to be present in the unit, Past usage, usefulness of input in their research. Possible future utilisation of your unit. Suggestions to improve the service as well as suggestions wishes for instruments/capability's currently not present. If you can get postgads promised to instruments and research programs including your facility the better. I know that is not always possible but if you do not ask you will not know. We did that at a previous institute where I was employed. This was a very useful exercise. This helped to direct the unit and improve on our service. These documents are also a very useful source for equipment motivation when it is combined into a format top management can understand.
Hopefully from this information the wise path to follow will be much more clearer to all.
Good luck.
-----Original Message----- } From: Rick Harris [mailto:raharris-at-ucdavis.edu] Sent: Friday, March 21, 2003 1:08 AM To: microscopy-at-sparc5.microscopy.com
After 24 years of managing our small facility as a collaborative effort with interested faculty, I now have a new supervisor with little technical expertise. Our facility consists of an older TEM, a new SEM, a new deconvolution scope, and a Lieca SP1 confocal. I don't want to air our dirty laundry but I need a little help in justifying our equipment to the new boss. Now, she is proud of the fact that she still uses PhotoShop 3.0 and states that the best way to get a good picture is to take a good picture and then don't mess with it. While there is certainly some truth in the second fact, it is also very naive. And the first fact classifies her as a Luddite. Her research centers around the confocal and the deconvolution scope. So the move is on to dump the TEM and give the Digital Darkroom to the department Network Admin. This will free me up to tackle confocal projects for her and spend more time dusting. Yes, after 24 years of managing the facility on my own my new boss has actually given me a dusting schedule. However, she is quick to point out that this is not micro-managing. Alright, I digress. I need to justify having a couple graphics workstations, a slide scanner, a flat bed scanner with transparency tray, and a photo-quality printer. She would give this equipment to the Network Admin who would integrate it with the common computer room. I treat this equipment like I do optical equipment. I am concerned that once relocated to a common room, with no supervision, it will rapidly deteriorate. I also take exception with the apparently wide-spread belief that the computer support staff is the best source of information on digital imaging. I guess because it is digital?
So how do I keep those pieces of equipment I deem necessary for the facility? I think we should have a photo-quality printer. I think we need the scanner with tray for those increasingly rare occasions that we need to scan in a TEM neg or an illustration for a PowerPoint slide or publication. I find that I am not good at making an argument for what seems blindingly obvious to me. I just sputter and fume and ask, "... how can anyone have an Imaging Facility without a couple workstations and printers?". Or a TEM.
And then there is always the possibility she is right. Maybe we don't need to do this stuff to the standards I embrace. Should I convert to her mantra: "Do more, less well."?
Any good arguments out there that have worked for any of you?
In the past I have thermally cycled several detectors without ill effect, but not enough times to be absolutely confident. Several vendors have told me that modern detectors should be far more stable to thermal cycling than older ones.
I have a detector from EDAX with a small cryostat designed for cooling on demand. This works fine, but we find the 1-2hr cool-down time is a nuisance in our facility for users who see a feature, then decide it would be good to do EDX analysis on it. We still tend to keep the detector cool except during the weekend. If you were more sure of when you would and wouldn't be using the EDX (and if the occasions when you would be using it are relatively few) then I would say this would be a good way to go. I have had one window failure (in 3.5 years), but then I have had window failures at about this rate with other detectors that are not cycled regularly.
If you are thinking of new technology, you might consider the silicon drift detector from Rontec. To the casual user this works just like a conventional EDX, but it only needs cooling to -15C or thereabouts; this is done easily with a thermoelectric cooler. Since the temperature change is small, it also cools fast if it is not already on - it can be used 15 mins or less after being switched on. On standby, the amplifiers are left on but the detector is allowed to warm up. I don't know if I got a particularly good one, but its resolution is comparable to that of a premium Si(Li) of a few years ago, and it is sensitive to x-rays down to C. It also will count happily at 160,000 counts/sec. It does have a few minor disadvantages, so it may not be for everyone, but I was attracted by the lack of need for Liq. N2, and I have been a happy customer. (I have no connection with Rontec except as a customer, but I should mention that the terms of the purchase allow Rontec to use my installation for demonstration purposes).
Cheers,
Tony.
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dear sir or madam, i was wondering whether you could inform me of the modern developments in microscopy and their advantages over traditional methods and machines
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It would seem this request has reached the ultimate in vague and general questions. I suppose the only improvement on it would be to avoid the mention of microscopy altogether.
At 02:41 PM 3/25/03 +0000, you wrote: } Date: Tue, 25 Mar 2003 14:41:04 +0000 } From: "R.M. Carr" {BMB2RMC-at-leeds.ac.uk} } Reply-To: noreply-at-leeds.ac.uk } Organization: University of Leeds } To: Microscopy-at-sparc5.microscopy.com } Subject: modern microscopy } Content-Type: text/plain; charset=us-ascii } Content-Transfer-Encoding: 7bit } } dear sir or madam, } i was wondering whether you could inform me of the modern developments } in microscopy and their advantages over traditional methods and machines } } -- }