Is there any standard data format for spectra of filters or fluorescent dyes for storage of transmission or absorption / emission values related to wavelength?
I would like to have spectra in a standard format which I can overlay for comparison of crosstalk, for checking suitability of combinations of filters or dichroics, fluorochromes, laser or arc lines etc.. Molecular probes as well as BioRad have dye or filter spectra available on their web sites, but only as GIF images etc. I am sure that manufacturers have general information at hand, but I think they do not want to distribute this information freely (at least not as tables).
What I imagine is a standard format as simple as possible (like ASCII tables) relating, e.g. wavelenght (in nm) to transmission, excitation or emission strength etc., and that such files can be imported and displayed by standard plotting software. Links to websites with standard spectra and related software would be helpful.
Thanks!
Guenter
---------------------------------- Dr. Guenter Giese MPI fuer Medizinische Forschung Jahnstr. 29 D-69120 Heidelberg Phone (Germany or 0-)6221-486-320 Fax (Germany or 0-)6221-486-325 e-mail: ggiese-at-mzf.mpimf-heidelberg.mpg.de ------------------------------------------
The correct adress is : hussein.jibaoui-at-univ-reims.fr
PhD in physics in search for a post doctoral. I will have finished my thesis on Sept 99. I'm actually working at Reims University (FRANCE) in the laboratory of analysis solid surfaces and interfaces. My research consists in exploration for new imaging technique X ray in total reflection (Application : topography observation of surface and interface solid/solid) and fluorescence X. In the team we use X ray microscopy in electrochimical systems. My competences : X ray microscopy : Projection, reflection and fluorescence Glancing X ray : Reletometry, TXRF, GIXF Experimentator data analysis preparation of thin layer by evaporation preparation of X-visible screen converter for image guide ........ Please send Email to receive my CV and details on my work. N.B : I will take part in XRM 99 conference to be held in Berkeley, California August 1-6, 99. Sincerly
I received the following inquiry today and wondered if any of you could help. Please reply directly to: Catrinel Stanciu [SMTP:stanciu-at-mbi-berlin.de] {mailto:[SMTP:stanciu-at-mbi-berlin.de]} as this person is not on the listserver.
-----Original Message----- } From: Catrinel Stanciu [SMTP:stanciu-at-mbi-berlin.de] {mailto:[SMTP:stanciu-at-mbi-berlin.de]} Sent: Wednesday, June 30, 1999 12:18 PM To: ebs-at-ebsciences.com {mailto:ebs-at-ebsciences.com}
Dear Microscopist,
I have an Indium Wire and is not aware of its use. It was come along the em kit. Could any one experience the use to it in electron microscopy.
I am also search for an e-mail address of Professor Willeur C. Bigelow.
Thanks in advance.
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road Pune- 411 004, India
Dear Catrinel, What is the wavelength of the laser emission? The choice of substance for the film will depend on the match between its absorption spectrum and the emission spectrum of the laser. Ideally, these should not overlap. An additional consideration would be that energy absorbed by the specimen could be transferred to (and thereby damage) the film, but this is likely to depend only on the mechanical strength of the film regardless of what it is made of, and anything suitable for HRTEM has to stand up under a similar process for energy absorbed from the elec- tron beam. Good luck. Yours, Bill Tivol
Boy I got a lot of responce on this one but........... It seems most people are using CD's. Our computer system people suggested NOT using cd r, or cd rw's stating that the hardware / software is still not standardised. Our falculty computer cluster has been using Zip (too small ?) and Jazz drives for years with no problems, and everyone seems to have one or both, and yes everyone has a CD too.
I will collect all the data and submit it to the server...and our computer "experts".
But I quess I still ask why are we storing their data, other than to have a repository of images for other uses? Can we use their images without their permission since it is the investigators research or do you (that allow outside use) have the investigator sign off rights on their photos?
Can we bring this topic up at the upcoming meetings? Thanks
You can buy polypropylene film from Goodfellow Corporation as thin as 4 microns. Chemplex sells it, too, but somene seems to have stolen my Chmeplex catalog.
Alternatively, my company MOXTEK has for years offered a service to refurbish CAMECA column seperation windows with our AP1 film, which is stronger, less permiable, and has higher nitrogen x-ray transmission than polypropylene.
best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
Yes, I would suggest the same storage medium, CD-R.
Remember, that CR-R can be sensitive to exposure to light for long times. So, in order to preserve them, you should not expose them to bright light (direct sunlight) for too long, or to too high a temperature. Storing them in a cabinet with doors should be adequate. Unlike magnetic media, CD-Rs don't care about magnetic fields.
Another option with higher capacity would be DVD. But I guess for writable DVDs you'll have to wait for a while.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Exchange Administrator Sent: Thursday, July 01, 1999 1:19 AM To: Michael Bode
At 02:40 PM 6/30/99 , you wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am having some trouble getting thin sections of some seeds I want to do EM immunolabeling on. I have ttried embedding some seeds in LR White (osmicated) or LR Gold (unosmicated). I took a week slowly infiltrating them with a dilution series of plastic resin but when I went to section them, they just popped out like stainless steel bb's. There is no hint at all that the embedding medium penetrated into them. Any experts out there with hints on fixing seeds?
TIA, Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
} I have an Indium Wire and is not aware of its use. It was come along the } em kit. Could any one experience the use to it in electron microscopy. }
Dear Rajdeep, Indium is used to make a vacuum seal for ultra-high vacuum applications. There should be a place where it would be used instead of an o-ring. This allows a vacuum seal without the use of grease or rubber, which can volatilize or outgas and degrade the vacuum. When you find the place for which the indium is intended to be a seal, place the indium so that it completely fills the seal volume and then some, so that when you tighten the seal the indium will be com- pressed. Indium is a soft metal and it will flow into the irregularities in the seal volume. Yours, Bill Tivol
Just a quick question to the wealth of knowledge on the microscopy list server,
How important is the WPE number on the Eponate 12 Resin bottle from Ted Pella when calculating the mixture of epon? i.e. should I recalculate the mixture every time the Eponate 12 Resin has a different WPE number???
Any help would be appreciated...
Eric A. Rosen Dept. Pathology UCLA Medical Center Electron Microscopy Lab...
} } I am having some trouble getting thin sections of some seeds I want to do } EM immunolabeling on. I have ttried embedding some seeds in LR White } (osmicated) or LR Gold (unosmicated). I took a week slowly infiltrating } them with a dilution series of plastic resin but when I went to section } them, they just popped out like stainless steel bb's. There is no hint at } all that the embedding medium penetrated into them. Any experts out there } with hints on fixing seeds? }
Dear Tom, We had similar problems with pollen grains. The trouble was with the difference between the hardness of the pollen and that of the resin. By varying the composition of the resin, we were able to make a sufficient match so that
sections could be cut--in our case, thick sections--with many--but not all-- of the pollen grains still in the sections. Good luck. Yours, Bill Tivol
The Tips & Tricks site has a few "difficult embedding" discussions archived. Go to:
http://www.biotech.ufl.edu/~emcl/index.html
follow the tips link and either run a search or manually pick through the TEM section. I am a year behind in archiving and will forward anything else I have to you but not to the list as a whole. If there is interest I will post the rest to the archive.
At 09:44 AM 7/1/1999 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I am interested in learning about sample preparation and previous work performed on dye adsorbed onto mica investigated via fluorescence microscopy.
Is it possible to demonstrate using fluorescence microscopy the charge anisotropy associated with the edges and faces of mica? How? Is there a pH dependence?
What literature is available that discusses fluorescence microscopy with
mica as a substrate?
Thank You for any tips or leads!
Christian Honeker Max Planck Institute for Polymer Research
I am trying to do immunolabeling of semithin sections of LR White (medium) blocks along with TEM of ultrathin sections.
I'm using the protocol for infiltration and embedding given by Brorson (Micron 1997, 1998): from 70% EtOH, 2 x 96% EtOH for 20 min., LR White/96% EtOH (1:1) for 2 hr, complete resin for 5 hr, embed in gelatin capsule, polymerize -at- 56 deg. C. ~40 hr
My questions are:
(1) I'm also using Brorson's protocol for immunolabeling. Brorson was immunolabeling TEM sections with gold-conjugated probes. I'm doing labeling of LM sections using peroxidase/DAB. My labeling has been weak at best. I'm going to try antigen retrieval with hot citrate buffer. Any other suggestions?
(2)My TEM sections have little holes. I can't think why; my tissue is fish heart muscle. This should be easy. My oven temp. is sometimes not very stable, but I can't see how this would cause holes. Should I try vacuum pressure during infiltration?
Thanks,
-- Sonia Cawsey McGowan Copley Library, University of San Diego email: scawsey-at-acusd.edu home page: http://www.acusd.edu/~scawsey
} -----Original Message----- } Boy I got a lot of responce on this one but........... } It seems most people are using CD's. Our computer system people } suggested NOT using cd r, or cd rw's stating that the hardware } / software is still not standardised. ...
I don't understand your "computer system people" ... that is, if the CDRW software offers you the option "write a disk which can be read by any platform" (?!?!?) ... even if they change the standard, new CD readers will always be able to read old standards ... they know if they do not, their drive will be outsold by another which does. Four years ago, we went with Fujitsu MOs ... they work beautifully, and we have 4 years of worked archived on them ... be we now have to admit they didn't catch on universally. We have since changed to CDs, but we'll not move the MO archives until we are down to our last MO drive, but all four of them are working as good as new. Regarding "zip drives" ... handy? yes ... an archive medium? NO! ... please do not trust magnetic media ... always go with optical or magneto-optical ..
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
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Hi Soumitra, Sorry your having problems. One thing you shouldn't do is dry your mount with anything, especially filter paper as there are some extractables which encourage further wetting. When your block gets wet from the boat just let it dry by evaporation. If you think the knife or block may have some contamination, wash them with a mild detergent and rince with hgih purity water and allow to air dry. Also never stop the block face near the knife edge as a static knife will likely wet. Keep the block face moving while it near the knife edge. Check to make sure water has not collected on the back edge of the knife as this can be invisible to you but will rewet your block face even though it appears dry Also make sure the water you are using is pure with no residuals. Good luck, Russ, Xerox
-----Original Message----- } From: Soumitra Ghoshroy [mailto:ghoshroy-at-nmsu.edu] Sent: Wednesday, June 30, 1999 10:42 AM To: Microscopy-at-sparc5.microscopy.com
Dear all microscopists,
I am having some problems during ultrathin sectioning with my block face getting wet as the diamond knife makes the first contact with the block and it happens with every stroke thereafter. I have dried the block face with filter paper or kimwipe before the next cutting stroke but it is not helping at all. Sometimes it happens during sectioning too. All my blocks are embedded in Spurr's and these are primarily plant tissue samples. I have switched knives, cleaned them before sectioning, lowered the water level in the boat, changed the clearance angle, but nothing has been working so far. I am not sure whether it is a lack of humidity problem or something else.
I will appreciate some help in this matter.
Thanks,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
I received the following inquiry today and wondered if any of you could help. Please reply directly to: Catrinel Stanciu [SMTP:stanciu-at-mbi-berlin.de] {mailto:[SMTP:stanciu-at-mbi-berlin.de]} as this person is not on the listserver.
-----Original Message----- } From: Catrinel Stanciu [SMTP:stanciu-at-mbi-berlin.de] {mailto:[SMTP:stanciu-at-mbi-berlin.de]} Sent: Wednesday, June 30, 1999 12:18 PM To: ebs-at-ebsciences.com {mailto:ebs-at-ebsciences.com}
Dear Microscopist,
I have an Indium Wire and is not aware of its use. It was come along the em kit. Could any one experience the use to it in electron microscopy.
I am also search for an e-mail address of Professor Willeur C. Bigelow.
Thanks in advance.
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road Pune- 411 004, India
I need to prepare TEM samples from a Fe-Co bulk material. Can someone suggest me an electrolyte for twin-jet electropolishing and the corresponding conditions?
Dear Tom, I've used a procedure covered by VA Lindley in Microscopy Research & Techniques 21:355-360 (1992) "A New Procedure for Handling Impervious Biological Specimens" with great success after I struck a similar problem with weevils (never had such a problem with Spurrs resin completely failing to stick to a sample). The paper recommends a pre treatment with gamma-glycidoxypropyl trimethoxysilane and subsequent embedding with LR White. Its simple & worked amazingly well with re-embedded weevils (they were irreplaceable samples so I stripped off the Spurrs). The paper mentions several samples which they tested the method on, including Salicornia oil seeds. I like it.
Good luck.
Regards,
Richard } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
Re: TEM, Phillips EM400 Good Morning Microscopy List Members With the kind help of the group, perhaps we will find certain salvaged parts for the Phillips EM400 TEM that we are in need of.... specifically: the Emission Chamber, the Electron Gun (Insulator and Wehnelt Assembly), the High Voltage Cable and the Vacuum Block (Upper Column). If anyone can help, we can be reached at T. (305) 669-0233 or email. Thanks. Stephen Quinto natural-immunogenics corp.
We are looking for Philips EM 400 series TEM in any condition, or at least for the following parts: emission chamber and upper column vacuum block, electron gun (insulator and Wehnelt assembly), and high voltage cable.
Indium wire is used as a vacuum seal in the column of Philips EM400 = (near the top of the liner tube, if my memory serves me right). This is = a one-use seal and has to be scraped out with a special tool supplied = for the purpose and replaced with new wire each time the vacuum seal is = broken.
-----Original Message----- } From: Rajdeep [SMTP:rajdeep-at-aripune.ernet.in] Sent: Friday, July 02, 1999 10:00 AM To: Microscopy-at-Sparc5.Microscopy.Com
Dear Microscopist,
I have an Indium Wire and is not aware of its use. It was come along = the em kit. Could any one experience the use to it in electron microscopy.
I am also search for an e-mail address of Professor Willeur C. Bigelow.
Thanks in advance.
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road Pune- 411 004, India
Shaf makes a good point in that CD's are platform independent. Most storage media suffer because the drive will eventually breakdown, at which point you will have to try to find an outdated drive to be able to access your archive.
We have been archiving on CD-R's for 5 years now. We started with Win 3.11, moved to Win 95, and are now using Win 98 & NT. All of the original discs are still readable, even ones written with the early CD-R software. We frequently give out data on the CD's and have had no problems with there accessibility on other computers.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: shAf {mshaf-at-darkwing.uoregon.edu} To: RLVAUGHN-at-UNMC.EDU {RLVAUGHN-at-UNMC.EDU} ; Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I have worked on EM equipment that uses indium wire as a vacuum seal. Indium allows you to make a vacuum seal between two flat surfaces. In the ADEM microscope we used it as a vacuum seal between the ceramic and stainless steel in the gun. If it was in your EM service kit, I would hang on to it.
Bill Carmichael Electron Microscopy Faculty Madison Area Technical College Madison, WI
http://electron-microscopy.madison.tec.wi.us
billc-at-jvlnet.com
Rajdeep 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 Microscopist, } } I have an Indium Wire and is not aware of its use. It was come along the } em kit. Could any one experience the use to it in electron microscopy. } } I am also search for an e-mail address of Professor Willeur C. Bigelow. } } Thanks in advance. } } Rajdeep Dongre } Electron Microscopy Laboratory } Agharkar Research Institute } G.G. Agarkar Road } Pune- 411 004, India
I'd like to get the information about supliers of the systems of cathodoluminescence detectors for SEM wich can include a monochromator to get a spectra of the radiation in the NIR range (400-1200nm). We know about Oxford system but it's a high end priced. Any suggestions will be appreciated. kind regards to all...
Krzysztof Herman * LABSOFT * Biuro Techniczno-Handlowe ul.Ba=BFancia 45A, 02-892 Warszawa PL tel/fax: (+48 22)6446233, tel: 6449750, 6449753 mobile: (+48 601)307456, (+48 501)213438 E-mail: kherman-at-labsoft.com.pl http://www.labsoft.com.pl
Hi Krzysztof - If you dont need a defined monochromatic image, a cheaper pan-chromatic imaging system used in conjuction with a fibre optic light guide as close as possible to the specimen, a vacuum feedthrough and an Ocean Optics SD2000 spectrometer does quite well. You can pick from a number of gratings for the appropriate wavelength range, but its probably a good idea to include the most sensitive. Ocean Optics have a good website, all the information is there. The only tricky bit is aligning the fibre optic - it has to be very precise.
Good luck
Sally Stowe ( no connection with any company, etc....)
Dr Sally Stowe Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475, ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525 http://www.anu.edu.au/EMU
I'd like to get the information about supliers of the systems of cathodoluminescence detectors for SEM wich can include a monochromator to get a spectra of the radiation in the NIR range (400-1200nm). We know about Oxford system but it's a high end priced. Any suggestions will be appreciated. kind regards to all...
Krzysztof Herman * LABSOFT * Biuro Techniczno-Handlowe ul.Ba=BFancia 45A, 02-892 Warszawa PL tel/fax: (+48 22)6446233, tel: 6449750, 6449753 mobile: (+48 601)307456, (+48 501)213438 E-mail: kherman-at-labsoft.com.pl http://www.labsoft.com.pl
I have received a request for TEM characterization of antiferromagnetic = nanocrystalline MnF2 powder. Is it okay to load the nanocrystalline = powder on to C coated copper grids / holey carbon film on copper grids? = I have used this method earlier for non-magnetic nanocrystalline oxide = ceramic powders. What will be the effect of the magnetic field of the = objective lens on the material? Is there a risk of contaminating the = objective lens pole pieces? What precautions should I take? I use a JEOL = 2000 EX II (T) and a Philips CM200.
Ideas / suggestions / advise requested, please.
Best wishes. --- R Divakar PMS, IGCAR, Kalpakkam 603102, India ----
I would like to know if any of you has had experience in TEM of block copolymers, specifically poly-(propylene-ethylene) impact copolymers? I use RuO4 to enhance the contrast between the rubber phase and the matrix. The RuO4 is prepared from the oxidation of hydrated RuO2 with NaIO4 (sodium periodate). I've tried using chlorosulphonic acid to harden the blocks before staining but realised that the acid was destroying both the particles and the matrix. I've sectioned both at RT and at cryo- temperatures but there is not much improvement. One of the main problems I'm facing is the poor penetration of the stain.
The literature that I've come across shows the structure to nice round particles in a semicrystalline matrix but my experience has shown me that there are more than two constituents and it's very rare that I come across rubber particles that are spherical with some crystallinity inside.
I would appreciate it if you could give me information as to the preparation of the solution, the specimen, the staining procedure, the ideal sectioning conditions, etc.
Sincerely Siyabonga Siyabonga Mange BSc (Pure and Applied Chemistry;UCT) MSc (Applied Science) Final year Materials Engineering Department Menzies Building, Level 2 UCT
Having spoken to someone who has used an In seal before, are there not problems with baking? In has a low melting point, so if you use it as a seal in any U= HV system that is baked, would you not be depositing In throughout the system?
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The =C5ngstr=F6m Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
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Sonia, A couple of things to think about: 1. What type of fixation are you using? That has great bearing on number of antigenic sites available for ICC. 2.Does your oven cycle over 60oC when in the heating phase? High temperatures can really effect the amount of labeling. It might be a good idea to lower the average temperature to 50oC so your chance of the oven overshooting is less. 3. 40 hours in the oven is fairly long....usually 24hrs is sufficient with LRW. 4. If possible, polymerize in a vacuum oven that allows you to back-fill with nitrogen. Keep vacuum low but on slightly to help eliminate any gas from within the sample. You can often find old paraffin ovens that can be used for this purpose. 5. How good is your antibody? Do you know that it will give a strong label? Not all antibodies are equal. What concentration are you using? 6. What size gold are you using? The larger the gold, the less actual particles will bind. 7. Do you have another antibody which you know labels well that can be used for a method control? Before you blame the prep (rather than the antibody) you should try to react your material with another antibody that you know is powerful and has a strong reaction and well defined localization pattern. 8. How about your background? If you have no background, even a weak label can be informative. 9. Pinholes in the resin may mean that too much water is being retained in the tissue. Try dehydrating through 100% ETOH and then make sure to keep samples in closed containers to minimize moisture uptake from the air.
ICC is not an exact science and there are often problems. Don't give up because when a reaction works, it can reveal really significant information. I have had numerous examples of an ICC reaction that didn't make sense (i.e. was different than our preconceived result expectations) which later led to months of intensive research using other techniques and important new conclusions.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Sonia Cawsey McGowan wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
by hermes.Teknikum.UU.SE (8.9.1a/8.9.1aa) with SMTP id QAA06354 for {microscopy-at-MSA.Microscopy.Com} ; Fri, 2 Jul 1999 16:56:07 +0200 (MET DST) Message-Id: {3.0.6.32.19990702163630.008f9e10-at-pop.teknikum.UU.SE} X-Sender: joba-at-pop.teknikum.UU.SE X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32)
Having spoken to someone who has used an In seal before, are there not problems with baking? In has a low melting point, so if you use it as a seal in any UHV system that is baked, would you not be depositing In throughout the system?
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The =C5ngstr=F6m Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
Hi Soumitra, sectioning can be very frustrating sometimes. Here are some solutions I have found to block wetting, in addition to the helpful suggestions of Rus Gillmeister: Are there any tiny nicks in the edges of your block face? They tend to draw water by capillary action. Make the edges of your pyramid very clean. Lowering the surface tention of the water is helpful, so you get a concave surface and can lead the water right up to the knife edge. You can do that with a drop of highly diluted Photoflo, but I have better luck with saliva. I use a toothpick that has been in my mouth to lead the water up to the knife edge, and it is just enough. The only problem is epithelial cells that sometimes show up in the 1-micron sections. However, they are no problem on thin sections, as they are not fixed, so they just disappear. During dry weather, a hot cup of coffee next to the ultramicrotome may be enough to humidify the air. I have even known people who put wet towels around their microtome. Anyway, good luck, and keep at it and it will get better. Honest. Joyce
Interesting. We have had occasions, where people with older CD-ROMs could not read CD-Rs created with a CD burner. Whether that is particular to a specific burner or a general problem I can't say. On the other hand it's a problem that can be solved by purchasing a new $50 CD-ROM.
But back to the original question of whether the facility has to keep images: Unless there are any laws or regulations requiring you to keep images as records, I would think you are free to set your own policy. I would set up a reasonable policy (like: we keep electronic copies for a year or two on CD. After that the CDs are discarded or sent to the user), let everybody know about this policy and deal with special circumstances one by one. I am pretty sure, that if you have a reasonable policy, 95% of your users will simply accept them.
When I worked with TEMs, we would give the users prints of the negatives but keep the negatives. of course once in a while somebody wanted to make a print of an old negative, but that was not too frequent.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Colin Reid[SMTP:CREID-at-TCD.IE] } Sent: Thursday, July 01, 1999 11:36:19 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Digital Archiving continued } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Shaf makes a good point in that CD's are platform independent. Most storage media suffer because the drive will eventually breakdown, at which point you will have to try to find an outdated drive to be able to access your archive.
We have been archiving on CD-R's for 5 years now. We started with Win 3.11, moved to Win 95, and are now using Win 98 & NT. All of the original discs are still readable, even ones written with the early CD-R software. We frequently give out data on the CD's and have had no problems with there accessibility on other computers.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: shAf {mshaf-at-darkwing.uoregon.edu} To: RLVAUGHN-at-UNMC.EDU {RLVAUGHN-at-UNMC.EDU} ; Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
For the past 16 years I've used an adhesion promotion step to embed difficult materials for diamond knife cross-sectioning. Typically hard, non porous materials such as glass, diamond, silicon, minerals, ceramics, semiconductors, metals, optical coatings and insect parts.
Prepare a 1% solution of Dow Corning's silane co-polymer Z-6040 (3-glycidoxy-propyltrimethoxy silane) in water and alcohol (50/50). Treat your samples in the solution for approximately an hour. Transfer them to filter paper to remove liquid and embed as normal in Spurrs or LR White. These coupling agents are the adhesion promoters used to bind fibers to polymer in fiberglass. Dow has a variety of other silane coupling agents with specificities for epoxies, acrylics, polyesters, phenolics, urethanes, etc. These are typically sold in 55 gal drums but smaller quantities are available.
Phil Swab Deposition Sciences Inc. Santa Rosa, CA 707-566-3718
-----Original Message----- } From: Tom Phillips [SMTP:PhillipsT-at-missouri.edu] Sent: Thursday, July 01, 1999 7:45 AM To: Microscopy-at-Sparc5.Microscopy.Com
I am having some trouble getting thin sections of some seeds I want to do EM immunolabeling on. I have ttried embedding some seeds in LR White (osmicated) or LR Gold (unosmicated). I took a week slowly infiltrating them with a dilution series of plastic resin but when I went to section them, they just popped out like stainless steel bb's. There is no hint at all that the embedding medium penetrated into them. Any experts out there with hints on fixing seeds?
TIA, Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Dear Greg Strout, John Warrack, Bill Tivol,Markus Meyenhofer,Mike Wombwell Mike Whittlesey and Jim Fotinopoulos, My sincerest thank you's to you all for your most invaluable help and for going out of your way to provide me the help I needed to resurrect this Polaron. Greg: a gracious thank you for going the "extra mile" to the point of sending me those beautiful images. What a help they are! It has been a pleasure conversing with you. John: those directions were extremely critical in assisting me with the carbon evaporating. I may contact you soon if I need further assistance. You are very kind. Bill: thank you for taking the time to provide such detailed and thorough oil changing instructions. You have this down to a science! Markus: It has been a pleasure speaking with you. Your help has truly been a benefit. Keep in touch. Mike W. and Mike W.: Thank you both for your valuable advise. Now I should be ready for the "maiden voyage". I will call you to further follow up. Jim: It was a pleasure to encounter your services. Your help was greatly appreciated. See you soon.
What a blessing this has been. Microscopists are truly a unique breed. I'm proud to be among the best.
Most Sincerely,
Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Hoechst Marion Roussel, Inc. 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-hmrag.com
} } Re: TEM, Phillips EM400 Good Morning Microscopy List } Members With the kind help of the group, perhaps we will find certain } salvaged parts for the Phillips EM400 TEM that we are in need } of.... specifically: the Emission Chamber, the Electron } Gun (Insulator and Wehnelt Assembly), the High Voltage Cable and the } Vacuum Block (Upper Column). If anyone can help, we can be reached } at T. (305) 669-0233 or email. Thanks. Stephen Quinto } natural-immunogenics corp. } }
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 530 754 7536 fax raharris-at-ucdavis.edu
Yes, underfill boat (concave surface), making sure knife edge is wet, but also decrease clearance angle and cut faster. When aligning, don't ever stop face even with the knife edge when it's really close.
Most important if the face is getting water on it is to trim the block with the glass knife so that scratches from trimming go parallel with the block bottom edge, rather than vertical. Trimming with a razor blade creates little troughs for the water to run right up onto the face. If you don't know how to do this, and want instructions, email back.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} -----Original Message----- } } ... } It seems most people are using CD's. Our computer system } people suggested NOT using cd r, or cd rw's ...
I thought I better post a caveat to my last message which would have implied CD/RW the only way to go. I've been spending all of this Friday trying to retrieve archived images from a CD written here. Bad writes ... not because there isn't any one standard format for acrchiving files to CDs across platforms, but because, either (1) I was cheap and bought inexpensive CDs, OR (2) because my CD writer needed its BIOS flashed for better support for silver CDs. A lesson learned ... buy best quality, "tried and true" gold CDs for your archiving purposes (!!!)
have a *happy* 4th ... shAf {} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Please respond directly, offline, to sbarlow-at-sunstroke.sdsu.edu
I am interested in hearing from vendors that specialize in converting older, analog Scanning Electron Microscopes to digital computer controlled microscopes.
I am also interested in vendors that specialize in SEM digital image capture
thanks.
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
All: I'm looking for some information on ultramicrotomy of materials, specifically semiconductors. I'm totally in the dark about it, except that I have heard the words. Book titles, articles, web sites, helpful hints, any information would be greatly appreciated.
Hi Eric - Put the blocks back into the oven; overnight at 80 C should harden them at bit better. Remember when the blocks come out of the oven they will be quite soft for a few minutes and take a couple of hours to reach maximum hardness. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Saturday, July 03, 1999 6:18 AM, ERIC [SMTP:biology-at-ucla.edu] wrote: } } } } Today over here at UCLA we are having a slight problem with soft blocks } trying to cut 1 micron sections first... } } any suggestions as to what to do to get the sections off the glass knife } and onto a glass slide?? } } The sections seem to get stuck on the edge of the knife after they have } been cut and are floating in the water but stuck to the knife edge?? } } Any help would be appreciated... } } Eric A. Rosen } Dept Pathology } UCLA Medical Center }
I would appreciate it if someone out there in the scientific community has information to offer on how the following can be used to make water safe for drinking (in terms of killing biological contaminants).
[1] sodium hypochlorite: Can one use household bleach (5.25% sod hypoch)? What dilution is effective and safe to use?
[2] potassium permanganate: What concentration to use?
Thank you for any help or suggestions where to find the info.
Hi Jonathan: A complex subject. The melting point of Indium is 156.61 degrees C. I expect none of the TEM lens parts get that hot during a bake out. Some manufacturers restrict In seals to a couple of positions, like the gun chamber. I do not have the figures, but believe that Indium, even near its melting point, when compared with good vacuum greases would outgas at much slower rate. Furthermore, a few indium metal atoms would not be contaminants which would cause problems similar to greases and oils and their degraded products. I believe to revert manufacturers In seals to polymers seals would be unwise. Comparing current instruments with those from the 70th, arguably cleaner vacuum systems have improved TEM performance most and In seals are part of that advance. The discussion should be if in turbo and ion getter pumped systems more In seals should be used in place of polymer seals. Disclaimer: PST supplies In seals - but also vac greases. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Saturday, July 03, 1999 12:37 AM, Jonathan Barnard [SMTP:Jonathan.Barnard-at-Angstrom.UU.SE] wrote: } } Having spoken to someone who has used an In seal before, are there not } problems } with baking? In has a low melting point, so if you use it as a seal in any } UHV } system that is baked, would you not be depositing In throughout the system? } } ******************************************************** } Dr Jonathan Barnard } } Analytical Materials Physics } The Angstrom Laboratory, Uppsala University } P O Box 534, SE-751 21 Uppsala, Sweden } Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 } } ******************************************************** } }
You don't have to use staining: our permanganic etching technique is ideal for PP-PE "impact" copolymers. You can see a picture of the result on:
http://www.reading.ac.uk/~spsolley/pp_impact.html
If you want any further information I can send it to you.
On Fri, 2 Jul 1999, Siyabonga wrote:
} I would like to know if any of you has had experience in TEM of block } copolymers, specifically poly-(propylene-ethylene) impact copolymers? } I use RuO4 to enhance the contrast between the rubber phase and the } matrix. The RuO4 is prepared from the oxidation of hydrated RuO2 } with NaIO4 (sodium periodate). I've tried using chlorosulphonic acid } to harden the blocks before staining but realised that the acid was } destroying both the particles and the matrix. I've sectioned both at } RT and at cryo- temperatures but there is not much improvement. One } of the main problems I'm facing is the poor penetration of the stain.
Whichever method you use, here's wishing you success,
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
we get good TEM specimens of PS/PE block copolymers if we first cut the material with the cryo-ultramicrotome (knife at -50degC, specimen at -140degC, 6deg cutting angle). Afterwards we stain the thin sections (on 1000mesh copper grids) with RuO4. Therefor we prepare a fresh mixture of about 100mg RuCl3 hydrate with ca. 5ml of 10wt% NaClO solution (in a vented hood of course!). Then we place the specimens over the mixture and keep them in the fumes for about two hours. Afterwards we wash them first with water and then with ethanol.
Good luck,
Petra
} Dear Colleagues, } } I would like to know if any of you has had experience in TEM of block } copolymers, specifically poly-(propylene-ethylene) impact copolymers? } I use RuO4 to enhance the contrast between the rubber phase and the } matrix. The RuO4 is prepared from the oxidation of hydrated RuO2 } with NaIO4 (sodium periodate). I've tried using chlorosulphonic acid } to harden the blocks before staining but realised that the acid was } destroying both the particles and the matrix. I've sectioned both at } RT and at cryo- temperatures but there is not much improvement. One } of the main problems I'm facing is the poor penetration of the stain. } } The literature that I've come across shows the structure to nice } round particles in a semicrystalline matrix but my experience has } shown me that there are more than two constituents and it's very rare } that I come across rubber particles that are spherical with some } crystallinity inside. } } I would appreciate it if you could give me information as to the } preparation of the solution, the specimen, the staining procedure, } the ideal sectioning conditions, etc. } } Sincerely } Siyabonga } Siyabonga Mange } BSc (Pure and Applied Chemistry;UCT) } MSc (Applied Science) Final year } Materials Engineering Department } Menzies Building, Level 2 } UCT } } Tel.:(021) 650 3181 (o/h) } (021) 387 4117 (a/h) } 083 583 8182 } Internet:SMANGE-at-ENGFAC.UCT.AC.ZA -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
I'm looking for some information on ultramicrotomy of materials, specifically semiconductors. I'm totally in the dark about it, except that I have heard the words. Book titles, articles, web sites, helpful hints, any information would be greatly appreciated.
Hi Becky,
Volume 31, No. 4 (July 1, 1995) of Microscopy Research and Technique was dedicated to ultramicrotomy of hard materials. In particular, there is a paper by S.R. Glanvill (page 275) on "Ultramicrotomy of semiconductors and related materials".
Best regards, Joergen
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
Just to add my own tricks to the excellent comments that you've received here, I would say that once you get the block face wet, drying it with a kimwipe never seems dry it enough for me. And one it's been wet, it has a tendency to get wet again much easier. So if I ever manage to get a wet block face (which happens less and less as you gain experience), I not only dry it with a kimwipe, but I blow dry the whole block face and knife edge with an air gun. (from one of those cans of compressed air) The diamond knife can stand drying from the air gun with no problem, and it's especially important to dry the back of the knife, and the block face.
Because after the block face gets wet once, then unless it is more than 100% dry after that, it will immediately get wet again and again and again....and slowly increase your frustrations until you require psychiatric treatment.
I want to do elemental mapping on catalyst particles with EELS. We have a Philips CM200 microscope equipped with a GATAN Image Filter. How does one prepare catalyst powders for EELS elemental mapping ? I suppose the powder has to be set in some kind of resin and then thinned by microtoming or some thinning technique to sufficient thickness. What kind of resin works best ? How thick should the sections be ?
Thanks
Willem Erasmus Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 9604211 Fax : +27 +16 9602826
Re: Ted Dunn's request for info on using bleach or potassium permanganate for purifying drinking water:
Household bleach (5.25% sodium hypochlorite) can be to disinfect drinking water for bacterial contamination. The amount you would use depends on the amount you are trying to disinfect. You should test the water 20 to 30 minutes after adding the bleach to check for the amount of free chlorine that is present in the water. You should have a free chlorine residule if 0.2 to 0.5 mg/L ( the greater the amount of contaminates in the water, the more chlorine it will tahe to reach this amount of residule chlorine). Kits can be purchased from various sources to measure the amount of chlorine present (the one I am familar with is made by Hach Chemical Co. but I am sure there are others available). These are similar to the kits used for measuring chlorene levels in swimming pools but test at lower ranges. Please be aware that this will not kill all pathogens. Those that are capable for forming cysts are an example and they can only be removed by filtering.
I do not know what the acceptable method is for using potassium permanganate.
If you are only wanting to purify small amount of water such as for use when backpacking, I would suggest you consider purchasing a hand held filter. These can remove all pathogens and are available at most local stores that carry camping and backpacking supplies.
Your best source of information is your local Boy Scout or camping store. We have used both sodium hypochlorite, iodine, and filtration. They each have their advantages & drawbacks. For cleaning biological out of water lines in a manufacturing setting (i.e., DI water), sodium peroxide is often used. For cleaning wells, dry sodium hypochlorite is used. The potassium permanganate is usually reserved for cuts and treating tropical fish.
Roy Nelson Material Testing Laboratory
DUNNTEM-at-aol.com"-at-sparc5.microscopy.com wrote: I would appreciate it if someone out there in the scientific community has information to offer on how the following can be used to make water safe for drinking (in terms of killing biological contaminants). [1] sodium hypochlorite: Can one use household bleach (5.25% sod hypoch)? What dilution is effective and safe to use? [2] potassium permanganate: What concentration to use?
Thank you for any help or suggestions where to find the info.
"When I were a lad", the British Army used to use a two tablet system: first one would add a "sterilizing tablet" to one's water bottle for a time, followed by a "thio tablet".
As regards the sterilizing tablet, to the best of my knowledge it was not hypochlorite as such, but something like Chloramine-T, the common name for
N-chloro p-toluene sulfonamide
which releases hypochlorite when dissolved in water. It is made from a by-product of saccharin manufacture. The "thio tablet" SOUNDS LIKE sodium thiosulfate, which would reduce any unreacted hypochlorite to chloride.
I would guess that these things might be on sale in a camping equipment shop.
On Sat, 3 Jul 1999 DUNNTEM-at-aol.com-at-Sparc5.Microscopy.Com wrote:
} I would appreciate it if someone out there in the scientific community has } information to offer on how the following can be used to make water safe for } drinking (in terms of killing biological contaminants). } } [1] sodium hypochlorite: Can one use household bleach (5.25% sod hypoch)? } What dilution is effective and safe to use? } } [2] potassium permanganate: What concentration to use?
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Willem Erasmus wrote: ============================================== I want to do elemental mapping on catalyst particles with EELS. We have a Philips CM200 microscope equipped with a GATAN Image Filter. How does one prepare catalyst powders for EELS elemental mapping ? I suppose the powder has to be set in some kind of resin and then thinned by microtoming or some thinning technique to sufficient thickness. What kind of resin works best ? How thick should the sections be ? ================================================= The ultimate goal is to have the sectioned sample "supported" in a way in which there are locations where the sample can be viewed without any support film. Also the section must itself be cut so it is absolutely of minimum thickness. For longer exposure times, drift can be minimized further through the use of higher grid mesh sizes (smaller hole sizes).
We have found that "supporting" the section on a holely carbon or Formvar ® film provides a stable section for viewing, with data being taken only from parts of the section that are suspended over a "hole". Hence the data is taken "neat" and without contribution (absorption) from a support film.
What resin to use? Vacuum embedding of at least some of the "Epon substitutes" (we have a natural preference for our own SPI-Pon™ 812 resin) gives quite an acceptable result. We use our own SPI "materials science" diamond knives but with 45° angles, not 55°. We have ourselves never been able to make thin enough sections with the 55° knives. 35° in theory would be even better but the knife will wear out much more quickly, perhaps too quickly for most budgets. Clearly other brands of diamond knives should work as well.
Just remember that section thickness is the critical issue and everything done must be from the perspective of minimizing that thickness (in order to minimize back ground effects).
Disclaimer: Our firms provides diamond knives and embedding resins and also performs sample preparation of this type as a service to others.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
A colleague at a local university is looking for a TEM of a centrisome for use in an in-house, intro biology lab manual. Does anyone have a photo they could donate (JPEG file is preferred)?
TIA
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (920) 42403404 wise-at-uwosh.edu http://www.uwosh.edu/departments/biology/wise/wise.html
A teacher has just contacted me with a request for help. Does anyone want to have an enjoyable, rewarding experience this fall? If you respond, please copy to me, so that I know that she's getting help.
} I currently teach the Talented and Gifted at my school, Findley Oaks Elem in } Duluth, GA (Atlanta Metro). When I ordered Microscopic Explorations: A GEMS } Festival Guide, I noticed the assistance your society provides as it relates } to volunteers, and obtaining microscopes. } } Our Talented and Gifted program focus is science, although we interweave all } the other disciplines. I am very much interested in meeting with one of } your members this summer to put some plans in place for the fall. Also, I } will be attending one of the GEMS workshop at the University of California, } Berkeley in August 1999. } } Any assistance you will provide is appreciated. } } Wanda Pennywell Rachal "MSPENNY" {MSPENNY-at-email.msn.com}
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Does anybody have experience (positive) with processing yeast for TEM, both conventional and rapid freezing - freeze-substitution? Favorite protocols, comments and tips would be much appreciated.
(I do thank greatly Scott Whittaker who has already directed me to some really good information on his tips website).
Sincerely, Vladislav V. Speransky School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Phone: 207 581 2998 Fax: 207 581 2969 Email: vladis-at-maine.maine.edu
Dear Willem, probably the simplest method to mount your partciles is to disperse them on a carbon holey grid using an organic solvent. I have seen others do this very successfully with ethanol for example. Put some of the catalyst in a test tube, add some ethanol shake to mix/disperse. Using a plastic pipette drop some of the mixture onto a carbon holey grid and let it dry. It should now be ready.
I hope this helps, Jonathan
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
I would like you to suggest the following problem:
I want to observe the arrangement of 2 proteins forming a ring shape- pore on Erythrocyte membrane (the ring shape is already observed by using TEM). The previous data suggests that they may form in to 6 sub unit -pore . I intend to use immunogold labelling 1 protein in order to adjust the interaction of two. But this pore is very small just about 3 and 7 nm inner and outer diameter respectively. So I am afraid of a steric hindrance and epitop exposure problem.
Could you please recommend which method and what kind of Microscope I should apply for my work?
Thank you in advance.
Nguyen Anh thi Van Lab. of Applied Microbiology Dep. of Biochemistry Faculty of Agriculture Tohoku University Sendai Japan
Caution re: If you are only wanting to purify small amount of water such as for use when backpacking, I would suggest you consider purchasing a hand held filter. These can remove all pathogens and are available at most local stores that carry camping and backpacking supplies.
FILTERS WON'T REMOVE VIRUSES, E.G., HEPATITIS A VIRUS.
On Mon, 5 Jul 1999 msteglic-at-notes.mdacc.tmc.edu-at-sparc5.microscopy.com wrote:
} Date: Mon, 5 Jul 1999 09:35:28 -0500 } From: msteglic-at-notes.mdacc.tmc.edu-at-sparc5.microscopy.com } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: potable water } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Re: Ted Dunn's request for info on using bleach or potassium permanganate for } purifying drinking water: } } Household bleach (5.25% sodium hypochlorite) can be to disinfect drinking water } for bacterial contamination. The amount you would use depends on the amount you } are trying to disinfect. You should test the water 20 to 30 minutes after adding } the bleach to check for the amount of free chlorine that is present in the } water. You should have a free chlorine residule if 0.2 to 0.5 mg/L ( the greater } the amount of contaminates in the water, the more chlorine it will tahe to reach } this amount of residule chlorine). Kits can be purchased from various sources } to measure the amount of chlorine present (the one I am familar with is made by } Hach Chemical Co. but I am sure there are others available). These are similar } to the kits used for measuring chlorene levels in swimming pools but test at } lower ranges. } Please be aware that this will not kill all pathogens. Those that are capable } for forming cysts are an example and they can only be removed by filtering. } } I do not know what the acceptable method is for using potassium permanganate. } } If you are only wanting to purify small amount of water such as for use when } backpacking, I would suggest you consider purchasing a hand held filter. These } can remove all pathogens and are available at most local stores that carry } camping and backpacking supplies. } } Mannie Steglich } Houston, TX } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I would like you to suggest the following problem:
I want to observe the arrangement of 2 proteins forming a ring shape- pore on Erythrocyte membrane (the ring shape is already observed by using TEM). The previous data suggests that they may form in to 6 sub unit -pore . I intend to use immunogold labelling 1 protein in order to adjust the interaction of two. But this pore is very small just about 3 and 7 nm inner and outer diameter respectively. So I am afraid of a steric hindrance and epitop exposure problem.
Could you please recommend which method and what kind of Microscope I should apply for my work?
ELECTRON MICROSCOPY TECHNICIAN, SENIOR Duties include all aspects of TEM, including negative staining and thin sectioning. It's an exciting job working with many different kinds of clinical and research specimens, physicians and researchers. It is a large lab with 4 other amiable EM techs who share duties. The salary is in the $27K range, plus 24% benefits (total--~$34.3K) which is reasonable for the cost of living here. Durham is a cosmopolitan city with 3 major universities nearby, and hence, all kinds of cultural entertainment; it is situated halfway between the beach and the mountains (about 3.5 hrs to each). Called the "City of Medicine," for it's high doctor/citizen ratio because of numerous hospitals, Durham also claims the team that "almost won" the NCAA tournament and that sent 3 players this year to the NBA (heaven help us next year). It is home to the Durham Bulls (baseball) and a half hour drive from the Carolina Hurricanes (hockey). With all this, it is still a relatively small city (~150K) where you can live in the country with only a 20 min drive to work, if you choose. Requisite job qualifications include experience in TEM, either through college courses or on-the-job training. Call or reply by email if interested.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Thanks to all of you who took the time to respond to my posting regarding block face getting wet. I received lots of very helpful suggestions.
This is truly an excellent resource for microscopists.
Cheers,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
Sample preparation required to perform EELS on catalyst particles varies, depending on the composition and hardness of the catalyst. We have good luck with many alumino-silicate and similar catalysts by embedding the finely ground powders in LR White resin (acrylic), and then microtoming to acquire very thin sections (references available). Many other supported catalysts do much better with an epoxy system such as Spurr's Low Viscosity resin. Some materials are easier examined by dispersing fine particles on a holey carbon support.
We'll need more info before we can give you any specific sample prep to follow. What is your catalyst material?
Good Luck, Brad ____________________________
I want to do elemental mapping on catalyst particles with EELS. We have a Philips CM200 microscope equipped with a GATAN Image Filter. How does one prepare catalyst powders for EELS elemental mapping ? I suppose the powder has to be set in some kind of resin and then thinned by microtoming or some thinning technique to sufficient thickness. What kind of resin works best ? How thick should the sections be ?
Thanks
Willem Erasmus Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 9604211 Fax : +27 +16 9602826
} Rick Vaughn wrote ... } } } It seems most people are using CD's. Our computer system } } people suggested NOT using cd r, or cd rw's ... } } I thought I better post a caveat to my last message which } would have implied CD/RW the only way to go. I've been spending } all of this Friday trying to retrieve archived images from a } CD written here. Bad writes ... not because there isn't any one } standard format for acrchiving files to CDs across platforms, } but because, either (1) I was cheap and bought inexpensive CDs, } OR (2) because my CD writer needed its BIOS flashed for better } support for silver CDs. A lesson learned ... buy best quality, } "tried and true" gold CDs for your archiving purposes (!!!)
Dear List:
All of this talk about digital archiving reminds me that film, that semi-transparent material with the silver halide coating, has a storage life of 100+ years when properly processed.
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 **********************************************
ELECTRON MICROSCOPY TECHNICIAN, SENIOR Duties include all aspects of TEM, including negative staining and thin sectioning. It's an exciting job working with many different kinds of clinical and research specimens, physicians and researchers. It is a large lab with 4 other amiable EM techs who share duties. The salary is in the $27K range, plus 24% benefits (total--~$34.3K) which is reasonable for the cost of living here. Durham is a cosmopolitan city with 3 major universities nearby, and hence, all kinds of cultural entertainment; it is situated halfway between the beach and the mountains (about 3.5 hrs to each). Called the "City of Medicine," for it's high doctor/citizen ratio because of numerous hospitals, Durham also claims the team that "almost won" the NCAA tournament and that sent 3 players this year to the NBA (heaven help us next year). It is home to the Durham Bulls (baseball) and a half hour drive from the Carolina Hurricanes (hockey). With all this, it is still a relatively small city (~150K) where you can live in the country with only a 20 min drive to work, if you choose. Requisite job qualifications include experience in TEM, either through college courses or on-the-job training. Call or reply by email if interested.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Thanks to all of you who took the time to respond to my posting regarding block face getting wet. I received lots of very helpful suggestions.
This is truly an excellent resource for microscopists.
Cheers,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
Our resident entomologist/electrophysiologist would like to delve into the fascinating world of microscopy and imaging, and wants to take me along with her.
My microscopy background has not included insects, and I am not well versed in their pecularities in preparation (such as how to deal with penetration of fixatives and the like through exoskeleton), so I am asking for your help with this.
At this point, I would very much appreciate it if people might point me in the direction of some good references and/or books on the subject of preparing cleared whole mounts of insects as well as techniques for fixing and embedding insect parts (especially antennae) for LM and EM.
Please reply off line. Thanks in advance.
Paula Allan-Wojtas Research Scientist, Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
} ---------- } From: Geoff McAuliffe[SMTP:MCAULIFF-at-UMDNJ.EDU] } Sent: Tuesday, July 06, 1999 9:32:49 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Digital Archiving continued
..
} Dear List:
} All of this talk about digital archiving reminds me that film, that } semi-transparent material with the silver halide coating, has a storage life } of 100+ years when properly processed.
.. and stored!! Exposure to light, humidity, chemical vapors, etc., or scratches due to handling can destroy negatives as well. I agree, that some negatives have a proven record of 100+ years, but how long do you relly need access to the negatives? I have folders and folders of TEM negatives from before the "digital age" in my shelves, and frankly, they will probably stay there until I die and then go to the trash without anybody ever looking at them again. And my filing system is not perfect either. If somebody asked me for a certain negative I took, say 10 years ago, I would probably have to look for a long, long time to find it. Some of the images went into publications, so there is another record of these. Others that I need I have on my PC and they will get transferred to other media until they become yesterday's news. Most of them are redundant and not needed, but since I had no way of making sure I recorded the right pictures I took many more than I needed.
While I agree, that negatives can survive at least a century, it is other factors like databasing, ease of transfer, and less work with chemicals that I would consider as well. The problem we have with recalling old data is usually not that the data is gone, but that the technology has changed and there is no longer access to the media (remember 8 inch disks :) ? I am convinced, that even in 50 years you can find somebody who can scrape the data from a CD and put it on a then current medium (holographic storage??).
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
I'm trying to get some information for a client who wants to try making his own glass Ralph (histo) knives. He says he heard something about a technique for breaking these by hand, without the special knifemaker. Does anyone out there in list-land know anything about this?
Thanks much.
Randy Tindall Electron Microscope Specialist Electron Microscope Core Facility University of Missouri Columbia, MO 65211
I'm trying to get some information for a client who wants to try making his own glass Ralph (histo) knives. He says he heard something about a technique for breaking these by hand, without the special knifemaker. Does anyone out there in list-land know anything about this?
Thanks much.
Randy Tindall Electron Microscope Specialist Electron Microscope Core Facility University of Missouri Columbia, MO 65211
Responding to the message of {v03007801b3a834474a66-at-[206.69.208.21]} from JoAnn Buchanan {redhair-at-leland.Stanford.EDU} : } } } Dear readers, I have a problem with very pale staining of my embed 812 } resin thin sections. I often need to use formvar and I cut 50nm thin } sections of cultured cells and fish embryos. I have tried numerous recipes } for both UA (including methanolic) and Lead (Sato's, lead nitrate, lead } citrate). It seems like the stain works well for a couple of times, then } flakes out. I have tried making the lead up fresh each time but even that } fails. The tissue is microwave processed including an en bloc uranyl } acetate step. I take great pains not to get any precipitate by using an } NaOH rinse. I stain 10-15 mins in UA and 5 min. in lead. Does any one have } any suggestions, ideas? What is the best lead stain?? Thanks in advance.
I, too, cut 50-55 nm sections of Embed 812, tho of plant materials, and also get slightly fainter staining, and I attribute it to just the fact that the sections are quite thin and there isn't a lot of material to stain - no Formvar on these grids. If I cut 60-75 nm sections, they do stain better. I'm cutting thin for better resolution.
Try cutting some thicker sections and look at those just to check your stains. I use 3-4% UA for 20-30 minutes, followed by Sato lead for 3-4 minutes. Hope this helps.
Gib
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu http://biosci.umn.edu/MIC/consortium.html
} Dear readers, I have a problem with very pale staining of my embed 812 } resin thin sections. I often need to use formvar and I cut 50nm thin } sections of cultured cells and fish embryos. I have tried numerous recipes } for both UA (including methanolic) and Lead (Sato's, lead nitrate, lead } citrate). It seems like the stain works well for a couple of times, then } flakes out. I have tried making the lead up fresh each time but even that } fails. The tissue is microwave processed including an en bloc uranyl } acetate step. I take great pains not to get any precipitate by using an } NaOH rinse. I stain 10-15 mins in UA and 5 min. in lead. Does any one have } any suggestions, ideas? What is the best lead stain?? Thanks in advance. } } JoAnn Buchanan } Molecular and Cellular Physiology } Stanford University Scholl of Medicine } Stanford, CA 94305
Sorry about the repeated posting about Ralph knives. The messages kept coming back to me as "undeliverable", so I didn't know they'd made it through. I thought the spam filter had locked me out. Oops...
Hello All Does anybody know where to buy following thing: - Thermal Insulating Compound type GE 7030 (pack 200 G) ... and what it is for ? kind regards Krzysztof Herman* LABSOFT * Biuro Techniczno-Handloweul.Bazancia 45A, 02-892 Warszawa PLtel/fax: (+48 22)6446233, tel: 6449750, 6449753mobile: (+48 601)307456, (+48 501)213438E-mail: kherman-at-labsoft.com.plhttp://www.labsoft.com.pl
} I'm trying to get some information for a client who wants to try making his } own glass Ralph (histo) knives. He says he heard something about a } technique for breaking these by hand, without the special knifemaker. Does } anyone out there in list-land know anything about this?
} Randy Tindall } Electron Microscope Specialist } Electron Microscope Core Facility } University of Missouri } Columbia, MO 65211
This is how I used to do it: 1) Make a score perpendicular to the long edge of a standard 1" strip of 1/4" plate glass, ~3" from the end. I did it with a LKB knifebreaker, but you could probably do it with by hand with a glass cutter. 2) Place a pencil under the score, on a table. Put a thinner stick under the 3" end piece (which will become the knife) to protect it from chipping. 3) Press firmly and evenly on both sides of the score; the glass will break. 4) Shorten the concave "ralph" to fit your knife holder. I used the LKB for this. You may have to experiment a bit with the position of the pencil if the cutting edge is too steep or shallow. This was actually published, a long time ago when ralphs were new, but I don't have the reference.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
} Can anyone give me their formula or direct me to a reference on how to } accurately calculate the size of scale bar on photomicrographs?
Easiest way to do this (for me, anyway) is as follows:
1. Think of a value for your scale bar, e.g. 1 micron (this should always be a round figure, or reasonable fractions thereof), 2. Multiply this by the final print magnification of the micrograph, 3. This value is the length of the scale bar, in this case in microns.
Example:
If final print magnification is 22 300x, and if you choose to have the scale bar representing 1 micron, then scale bar length = 1 micron x 22 300 = 22 300 microns. Then, as there are 1000 microns in a mm, the scale bar length on the micrograph will have to be 22.3mm.
Clearly, depending on the magnification, sometimes you may choose an impractical figure for the scale bar to represent.
If, for example you choose 1 micron for a magnification of 400x then the scale bar would only be 400 microns (0.4mm) long. This is crazy so how about choosing 20 microns? The scale bar would then have to be 20 microns x 400 = 8000 microns = 8mm.
At the other end of the scale, if you have a magnification of 120 000x then a 1 micron scale bar would be 120mm - much too long - so how about a 0.1 micron (or 100 nm) scale bar? Yes, much better because then the scale bar would be 0.1 micron x 120 000 = 12 000 microns = 12mm.
PLEASE do not fall into the trap of doing it the other way around and calculating what a bar of given length will represent. This leaves you then with all your bars the same length (very neat, yes) but representing crazy figures like 0.23 micron, 66 nm, etc!
Thanks for asking about this. Now, having written out the answer, with examples, on the many occasions when I am asked the same question by our users, I can simply pass on copies of this message rather than do the explanation over and over again on the board!
Good luck
Robin
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za (or eurc-at-giraffe.ru.ac.za) tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
Regarding your closing sentence - I shall keep your letter for 50 years and see if you were right !! Either way, I know that my indexed collection of negatives will still be around ! Predicting the future of technology at this exciting time is a high-risk operation - even for 10 years never mind 50.
Tony Bruton Centre for EM University of Natal, KZN South Africa
} } } Michael Bode {mb-at-soft-imaging.com} 07/06 10:39 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ---------- } From: Geoff McAuliffe[SMTP:MCAULIFF-at-UMDNJ.EDU] } Sent: Tuesday, July 06, 1999 9:32:49 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Digital Archiving continued
.
} Dear List:
} All of this talk about digital archiving reminds me that film, that } semi-transparent material with the silver halide coating, has a storage life } of 100+ years when properly processed.
. and stored!! Exposure to light, humidity, chemical vapors, etc., or scratches due to handling can destroy negatives as well. I agree, that some negatives have a proven record of 100+ years, but how long do you relly need access to the negatives? I have folders and folders of TEM negatives from before the "digital age" in my shelves, and frankly, they will probably stay there until I die and then go to the trash without anybody ever looking at them again. And my filing system is not perfect either. If somebody asked me for a certain negative I took, say 10 years ago, I would probably have to look for a long, long time to find it. Some of the images went into publications, so there is another record of these. Others that I need I have on my PC and they will get transferred to other media until they become yesterday's news. Most of them are redundant and not needed, but since I had no way of making sure I recorded the right pictures I took many more than I needed.
While I agree, that negatives can survive at least a century, it is other factors like databasing, ease of transfer, and less work with chemicals that I would consider as well. The problem we have with recalling old data is usually not that the data is gone, but that the technology has changed and there is no longer access to the media (remember 8 inch disks :) ? I am convinced, that even in 50 years you can find somebody who can scrape the data from a CD and put it on a then current medium (holographic storage??).
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} A complex subject. The melting point of Indium is 156.61 degrees C. I expect } none of the TEM lens parts get that hot during a bake out. Some manufacturers } restrict In seals to a couple of positions, like the gun chamber. I do not have } the figures, but believe that Indium, even near its melting point, when } compared with good vacuum greases would outgas at much slower rate.
Since the boiling point of Indium is 2000 C plus change (at 1 atmosphere pressure), the vapor pressure would be very low. I don't have a published value.
} Furthermore, a few indium metal atoms would not be contaminants which would } cause problems similar to greases and oils and their degraded products.
Agreed. They'd occasionally be scrubbed off surfaces by stray radiation, but would otherwise sit harmlessly on the column.
} The discussion should be if in turbo and ion getter pumped systems more In } seals should be used in place of polymer seals.
I prefer systems like conflat flanges with copper gaskets. They're easier to work with and have equally good vacuum properties. Yours, Bill Tivol
I have made some of these for large cutting edges on a rotary microtome, but never for the triangular pieces that are typically used in an ultratome. I used the standard 1/4" thick LKB glass and scored it with a diamond scribe, at right angles to the length of the glass strip -using a staight edge to guide the cut. Then I broke the strip with glass-breaking pliers (check the hardware store). The same method can be used for making ralf knives from 3X1 microscope slides (the cheaper the glass the better they break---I don't know why). It takes a few tries to get the edge that you want. As I said, I only make square or rectangular knives this way, 'tho you may be able to work something out for the triangular type.
Karen
On Tue, 6 Jul 1999, Tindall, Randy D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I'm trying to get some information for a client who wants to try making his } own glass Ralph (histo) knives. He says he heard something about a } technique for breaking these by hand, without the special knifemaker. Does } anyone out there in list-land know anything about this? } } Thanks much. } } } Randy Tindall } Electron Microscope Specialist } Electron Microscope Core Facility } University of Missouri } Columbia, MO 65211 } } }
I have found it quite useful to print out an Excel spreadsheet table with values on it. One axis can have magnifications in 1,000s (eg 1,2,3,4,5,6,7,8,9,10,20,30,40,50,60,70,80,90,100,200,300,400,500) and the other axis can have fixed values to be represented (eg 10,20,50,100,500nm;1,2,5,10,20um). The table can then contain the values in mms.
It's almost foolproof and easy to run off copies for students, especially when they realise that you can add bits together on the magnification side to give you a scale bar length for say 13k = 10k + 3k so just add the two entries in the table to get the length for a 2um bar. I imported this into a word document because I wanted to shade bits of the table to give recommended lengths for scale bars but it works fine.
I may have the Word 6.0 for Windows PC document somewhere. If you want a copy post an e-mail to me (see e-mail address at bottom)
Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: Robin Cross To: Ruth Yamawaki Cc: microscopy
Hello Ruth
} Can anyone give me their formula or direct me to a reference on how to } accurately calculate the size of scale bar on photomicrographs?
Easiest way to do this (for me, anyway) is as follows:
1. Think of a value for your scale bar, e.g. 1 micron (this should always be a round figure, or reasonable fractions thereof), 2. Multiply this by the final print magnification of the micrograph, 3. This value is the length of the scale bar, in this case in microns.
Example:
If final print magnification is 22 300x, and if you choose to have the scale bar representing 1 micron, then scale bar length = 1 micron x 22 300 = 22 300 microns. Then, as there are 1000 microns in a mm, the scale bar length on the micrograph will have to be 22.3mm.
Clearly, depending on the magnification, sometimes you may choose an impractical figure for the scale bar to represent.
If, for example you choose 1 micron for a magnification of 400x then the scale bar would only be 400 microns (0.4mm) long. This is crazy so how about choosing 20 microns? The scale bar would then have to be 20 microns x 400 = 8000 microns = 8mm.
At the other end of the scale, if you have a magnification of 120 000x then a 1 micron scale bar would be 120mm - much too long - so how about a 0.1 micron (or 100 nm) scale bar? Yes, much better because then the scale bar would be 0.1 micron x 120 000 = 12 000 microns = 12mm.
PLEASE do not fall into the trap of doing it the other way around and calculating what a bar of given length will represent. This leaves you then with all your bars the same length (very neat, yes) but representing crazy figures like 0.23 micron, 66 nm, etc!
Thanks for asking about this. Now, having written out the answer, with examples, on the many occasions when I am asked the same question by our users, I can simply pass on copies of this message rather than do the explanation over and over again on the board!
Good luck
Robin
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za (or eurc-at-giraffe.ru.ac.za) tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
by darkwing.uoregon.edu (8.9.3/8.9.3) with SMTP id IAA26704; Wed, 7 Jul 1999 08:05:27 -0700 (PDT) "Ruth Yamawaki" {yamawaki-at-leland.Stanford.EDU} Cc: {microscopy-at-Sparc5.Microscopy.Com}
Robin Cross writes ...
} -----Original Message----- } } Easiest way to do this (for me, anyway) is as follows: } } 1. Think of a value for your scale bar, e.g. 1 micron } (this should } always be a round figure, or reasonable fractions thereof), } 2. Multiply this by the final print magnification of the } micrograph, } 3. This value is the length of the scale bar, in this } case in microns. } } Example: } ...
Indeed the simplest approach ... however, when I first read Ruth's post I hesitated to reply without wanting more info. Is she asking about creating a ubar after darkroom enlargement? ... or is she asking about converting magnifications into pixel dimentions for annotating with (e.g.) Photoshop?? Or, is she asking an even more basic question which might be instrument specific and about how to calibrate "reported" magnifications???
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I admit that it is dangerous to speculate on technological advances and extrapolate too far into the future, but it goes both ways. My daughter just showed me a book from the 60s (I believe), where they predict, that "by the year 2000 vacation travel to the moon will be commonplace", and we have all been there, right?
The reason I used CDs for my "prediction" is that it is used everywhere - data distribution, Music, software, etc. - and it will have a pretty big momentum to be totally replaced and forgotten.
I still have an old PC with a 5 1/4 floppy around (use it as a doorstop, mainly), but I could probably revive it to read 5 1/4 disks and put the data on CDs, if that became necessary. Those disks were around in the early 70s into the 80s, so they are 15 to 20 years old. And at that time, there were much fewer PCs around than now. So, if I personally can go back 15 to 20 years with a 5 1/4 floppy, chances are that somebody could go back to CDs in 50 years and read them, particular since CDs are much more widespread than 5 1/4 floppies ever were.
But let's discuss this in 50 years over a glass of wine or beer, perhaps at the 2049 M&M meeting (on Mars???)
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Tony Bruton[SMTP:BRUTON-at-EMU.UNP.AC.ZA] } Sent: Wednesday, July 07, 1999 3:59:04 AM } To: Michael Bode; Microscopy-at-sparc5.microscopy.com } Subject: RE: Digital Archiving continued } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Micheal
Regarding your closing sentence - I shall keep your letter for 50 years and see if you were right !! Either way, I know that my indexed collection of negatives will still be around ! Predicting the future of technology at this exciting time is a high-risk operation - even for 10 years never mind 50.
Tony Bruton Centre for EM University of Natal, KZN South Africa
} -----Original Message----- } } Regarding your closing sentence - I shall keep your letter for 50 } years and see if you were right !! Either way, I know that } my indexed } collection of negatives will still be around ! ...
I don't believe anyone is asking you to trash your collection of negatives ... which (no arguement from anyone) IS the best and least expensive method of archiving photographic images ... IF they're on film!! On the other hand ... what if, for their own reasons, one of your students chooses to scan all of one your past project's micrographs, and turns them and their analysis into a thesis project. Further imagine some squirrel chooses to fry itself on your building's power transformer while that computer is on and it wipes out a sector on the harddisk, and with it all the work he/she has done. I wonder how appreciative your student will be that your negatives are so well indexed(???) ... my $0.02 :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I saw someone post microwave antigen retrieval protocol website a few days ago in Listserver. However, I deleted it by accident. Could anyone forward to me the website or any other resources under that subject. I try to use microwave to do immunohistochemistry. Thanks.
******************************************************************* To see what is in front of one's nose requires a constant struggle.
George Orwell
Dorothy Zhang Harvard School of public Health Building 2, CVLAB 677 Huntington Ave, Boston, MA 02115 Phone# 617-432-6981 Fax# 617-432-2980
-----Oorspronkelijk bericht----- Van: Tindall, Randy D. {TindallR-at-missouri.edu} Aan: 'microscopy-at-sparc5.microscopy.com' {microscopy-at-sparc5.microscopy.com} Datum: mercredi 7 juillet 1999 5:15 Onderwerp: histo knives
[knip]
A method for "do-it-yourself" glas knives to cut semi-thin sections (about .5 - 3 micron) of specimens embedded in methacrylate, Epon and/or Araldite is described in:
Gerlach, D: "Botanische Mikrotechnik", Thieme, stuttgart (1969), ISBN unknown, the method is also described in more recent versions of the book, but I don't have these at hand at the moment... (in German, maybe there's an English translation...).
The method described uses a regular rotary microtome with do-it-yourself modification of the knive holder. No appartus is needed to make the glass knives.
Does anyone know of an antibody source for GFP(not produced in rabbit) = that has a proven track record for EM immunolabelling? This will be used = in a dual labeling experiment in which the other primary is produced in = rabbit.
Thanks, Hank Adams IMC Baylor College of Medicine Houston, TX
Hello again I'm just full of questions lately. As I mentioned in a previous question, we are in the middle (actually down to the wire) of a CAP inspection in the Path EM lab. We are having to rewrite all of the procedures which most have been done from memory and little black books in our lab coats. Sure the originals are there but the modifications aren't. One of which is a notation not to heat the paraformaldehyde solution greater than 60 degrees C, and I remember the senior lab tech at the time telling me this was to prevent polymerization. Unfortunately, the current books like Bozzolla and Russell"s 2nd edition state that you can heat it from 60 to 80 degrees. Are these temperatures OK, I know it would probably dissolve faster? Was this "polymerization" stuff hooey? Thanks for the help.
Rick Vaughn, M.S. Electron Microscopy Research Fac. Dept. Cell Biology and Anatomy Univ Neb Med Ctr
PS....I say "we" but the real work is being done by the new lab coordinator. I'm luckily just the technical adviser that has been around for 12 years.
shAf, Robin's reply was what I wanted. Thanks to both of you for replying.
Ruth
At 08:05 AM 7/7/99 -0700, shAf wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We use TWAIN import feature in Photoshop to bring images from two cameras on two stereomicroscopes. We have John Russ' Image Processing Toolkit for Photoshop. There is one plug-in, "IP*measure-Calibrate" that can be used to calibrate the image if a feature of known dimension is captured. What I did was to take a digital image from a good metric ruler at each magnification setting of the microscope. I then calibrated the image, drew a scale marker on a new layer and labeled it for each setting. I changed the name of each layer to be indicative of the setting on the microscope that changed the mag. The only thing on each of these layers is the scale marker and the label. Once I had a layer for each of the different settings, I gathered them all into one photoshop file and labeled it with the appropriate microscope name. When I want to capture images from the microscope, I open Photoshop by opening this file of calibrated layers. After I collect the new image, I drag the appropriate layer from the open "calibrated scale markers" file onto the new image and align the scale marker where I want it.
A little work up front has saved me a lot of aggravation when it comes to calibrating images and putting scale markers on images. I plan to do the same thing for the mag settings on my TEM when I digitize them with my negative scanner, but I haven't invested the time yet.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Does anyone have any opinions on using a heat pen to flatten ultrathin EM sections? I would very much prefer to get away from using chloroform.
Electron Microscopy Sciences has two models: Wax Pen 1 (which uses 1 AA battery) and Wax Pen 2 (which uses 2 AA batteries). Their tech representative recommended the Wax Pen 2 for flattening sections. Is this consistent with the preference of the majority?
I need to decide to place an order soon (the end of the fiscal year is approaching).
Thank you,
Jaclynn Lett, Research Assistant jmlett-at-cid.wustl.edu
Fay and Carl Simon Center for the Biology of Hearing and Deafness Central Institute for the Deaf 818 S. Euclid Ave. St. Louis, MO 63110
We have two CD Writers here. They are both Plasmon RF4100 units. They are five & fours years old respectively. Apart from a dodgy power supply on the older one they have given very reliable service since we got them. There is bound to be a much more modern version available. We were recommended to purchase the first one because it was a more professional model compared to others available. The "engine" is a Philips. It might be worth trying the Hewlett P model since it may be more readily available and service will be more local.
When considering the model to purchase they will quote Write/Read speeds. The computer you use to write will determine what Write speed you are able to use. The software you use should give a reading of the data transfer & more especially the level of data in the buffer ( I use Gear 4 ). If the buffer empties because of a fast Write speed then the CD will be ruined. I have two computers set up for writing. One cannot reliably Write faster than X1, while the second writes at X2 successfully. Both are set up with SCSI transfer from a SCSI hard drive. The Read speed is unimportant since I just pop it into the X32 CD on the computer when I want to check it.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2. Tel: 353-1-6081820 Fax: 353-1-6770438 Email: creid-at-tcd.ie http://www2.tcd.ie/Electron_Microscope/emu/home.htm
-----Original Message----- } From: rlvaughn-at-UNMC.EDU [SMTP:rlvaughn-at-UNMC.EDU] Sent: Thursday, July 08, 1999 2:17 AM To: sdw-at-biotech.ufl.edu; oshel-at-terracom.net; gary-at-gaugler.com; creid-at-tcd.ie; m.dickson-at-unsw.edu.au; walck-at-ppg.com; raharris-at-ucdavis.edu; doug-cromey-at-ns.arizona.edu; mshaf-at-darkwing.uoregon.edu; mb-at-soft-imaging.com
I think you'd do better with a calibrating slide on your microscope. It's accurate and can be used with any objective lens. The one I know of is made by MicroBrightField (www.microbrightfield.com) and shows two different sized 10x10 square lattices, one for high mag objectives, the other for low. You can also make video images of the lattices. I believe the individual box sizes are 20 um and 100 um. The price is modest, somewhere around $100.
Ed Glaser
On Wed, 7 Jul 1999, Walck. Scott D. 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 use TWAIN import feature in Photoshop to bring images from two cameras on } two stereomicroscopes. We have John Russ' Image Processing Toolkit for } Photoshop. There is one plug-in, "IP*measure-Calibrate" that can be used to } calibrate the image if a feature of known dimension is captured. What I did } was to take a digital image from a good metric ruler at each magnification } setting of the microscope. I then calibrated the image, drew a scale marker } on a new layer and labeled it for each setting. I changed the name of each } layer to be indicative of the setting on the microscope that changed the } mag. The only thing on each of these layers is the scale marker and the } label. Once I had a layer for each of the different settings, I gathered } them all into one photoshop file and labeled it with the appropriate } microscope name. When I want to capture images from the microscope, I open } Photoshop by opening this file of calibrated layers. After I collect the } new image, I drag the appropriate layer from the open "calibrated scale } markers" file onto the new image and align the scale marker where I want it. } } A little work up front has saved me a lot of aggravation when it comes to } calibrating images and putting scale markers on images. I plan to do the } same thing for the mag settings on my TEM when I digitize them with my } negative scanner, but I haven't invested the time yet. } } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } } "The opinions expressed are those of Scott D. Walck and not of PPG } Industries, Inc. nor of any PPG-associated companies." } }
Edmund Glaser, D. Eng. Dept. Physiol. Univ. Md. School. Med. Baltimore, MD 21201 USA Ph: (410) 706-5041 Fax: (410) 706-8341
Hello, We need to reduce effects of acoustic interference with high resolution EFTEM work. Would anyone be willing to share experience with effectiveness of various wall panels, carpets, or floor tiles for this purpose? Marek.
Marek Malecki, M.D., Ph.D. Director Electron Microscopy Facilities
Dear All, Is anyone aware of a reference that will give the DeBye Waller Factors of Zn and O in ZnO? Or do you know of a value from a private communication or experiments ? Aacknowledgements will be made to those providing the information. I would really apprecite any help on this one, thanks, Jonathan
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
Dear Rick: Yes, I think it is OK to heat paraformaldehyde above 60 degrees. We have used 1% paraform in combination with 2% glutaraldehyde for perfusion fix. The source of our recipe is fuzzy. Try Karnovsky, M.J., 1965. J. Cell Biol. 29:137A. Our recipe is as follows:
1) 1 gram PF powder in 50ml DD H20. 2) Stir and heat to 70 degrees C 3) Transfer to stirrer (cool) 4) Add 2 drops 1N NaOH by pasteur pipette 5) Solution should clear. 6) Filter thru #1 filter paper.
This gets diluted by half when combined 1:1 with 4% glut. Don Gantz Boston Univ Med School gantz-at-med-biophd.bu.edu
It's a while since I last cut thin sections, but I always used to use a heat pen. I bought it when there was only one model and it ran off the mains through a transformer, but I imagine the present ones are much the same. Anyway, I found it worked very well, and it was wonderful not to breathe in chloroform all the time.
Lesley Weston.
On Wed, 7 Jul 1999, Jaci Lett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have any opinions on using a heat pen to flatten ultrathin EM } sections? I would very much prefer to get away from using chloroform. } } Electron Microscopy Sciences has two models: Wax Pen 1 (which uses 1 AA } battery) and Wax Pen 2 (which uses 2 AA batteries). Their tech } representative recommended the Wax Pen 2 for flattening sections. Is this } consistent with the preference of the majority? } } I need to decide to place an order soon (the end of the fiscal year is } approaching). } } Thank you, } } Jaclynn Lett, Research Assistant jmlett-at-cid.wustl.edu } } Fay and Carl Simon Center for the Biology of Hearing and Deafness } Central Institute for the Deaf } 818 S. Euclid Ave. } St. Louis, MO 63110 } } voice 314-977-0257 fax 314-977-0030 } } } }
We are looking to identify any local or regional testing labs or vendors that could help us evaluate the feasibility of applying laser confocal microscopy applied to some of our coatings and coatings-related studies. Please respond directly. Thank you.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
} I don't believe anyone is asking you to trash your collection of } negatives ... which (no arguement from anyone) IS the best and least } expensive method of archiving photographic images ... IF they're on } film!!
If you are saying that sticking existing negatives in a box and index them somehow (on paper or computer), you'll get no argument from me. However, if you are saying that taking images on film and then archiving them is the least expensive way, I would argue otherwise. The negative itself is roughly $1/piece (perhaps a bit cheaper if you buy bulk). So, the cost for 300 images is about $300 without counting development chemicals and labor. I also leave out initial investments (darkroom, development station, other equipment, enlarger, etc.). The same 300 image you could probably stuff onto one (1) CD for about $1. Again I am neglecting initial investments (CD-writer, camera, PC, etc), but I would say, the cost difference points in the direction of digital imaging.
Of course I will now get a lot of arguments about prices of cameras vs. enlargers, upgrade costs, labor cost, information density on film vs. CD, etc. The decision, which is more cost effective has to be made individually, taking into account the number of images recorded on average, existing equipment, etc., but just saying "Film is cheaper" is not enough. It needs to be qualified.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
This is the Biological EM Facility, but, with the recent acquisition of a LEO 912 EFTEM, I suddenly find non-biologists at my door! There promises to be some exciting projects.
One researcher is trying to visualize and characterize Silicon nanoparticles. They may or may not be crystalline, and they may or may not form crystals and/or amorphous blobs. The immediate question (for a grant proposal) is what do we need to do electron diffraction? We can do s.a.d., but I'm completely unclear as to the other kinds of diffraction and what information can be obtained. His particles are probably 1-4 nm, with another class at probably 6-12 nm. I read somewhere that low-angle diffraction is probably useful only on particles {0.4 nm. S.a.d. probably for larger particles. What would people recommend? And right now I could use a primer/glossary on CBED, SAD, HRED, etc.
I love learning new techniques, but I never thought I'd really have to know what Kikuchi lines were!
Aloha from a biologist, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Does anybody out there know where I can find a used liquid nitrogen dewar? Somewhere around 4 liter size would be fine. My telephone number is 202-544-0836 and e-mail is mansfield-at-erols.com
Our technicians and students in the TEM course routinely use the heat pen for flattening thin sections. We have been using the "Max Wax" pen marketed by EMS. We use rechargeable nickel-metal hydride batteries that last longer than the regular nickel cadmium ones. We also keep extra tips for the pens (also avalibale from EMS) that needs replacement depending on the amount of use.
RESEARCH SCIENTIST: Microcosm, Inc., one of the top photonics imaging laboratories in the world has an immediate opportunity in Columbia, Maryland for a Research Scientist specializing in Time-Resolved Fluorescence Imaging and Photonics. You should have an advanced degree in physics, biophysics, material science or a related area with 0-3 years postgraduate laboratory experience (Ph.D.) or 3-5 years experience (MS degree). Essential skills include experimental design, execution and troubleshooting, and employing methods such as multiphoton excitation, time-resolved spectroscopy and imaging. Strong analytical, computer and verbal skills are required. We offer an attractive salary, comprehensive benefits and progressive work environment. This position is open only for US citizens.
Please fax or e-mail CV to Dr. H. Malak at 301-725-2941 or henryk-at-microcosm.com
________________________________________________________ Dr. Henryk Malak Director of Research Microcosm, Inc. | 9140 Guilford Road, Suite O |Columbia, MD 21046 Phone: (301) 725-2775, Fax: (301) 725-2941 Our web site is located at: http://www.microcosm.com {http://www.microcosm.com} ________________________________________________________
This is the Biological EM Facility, but, with the recent acquisition of a
} LEO 912 EFTEM, I suddenly find non-biologists at my door! There promises } to be some exciting projects.
You will also have some exciting biological projects--that's a terrific machine for getting the maximum information from each electron.
} } One researcher is trying to visualize and characterize Silicon } nanoparticles. They may or may not be crystalline, and they may or may not } form crystals and/or amorphous blobs. The immediate question (for a grant } proposal) is what do we need to do electron diffraction? We can do } s.a.d., but I'm completely unclear as to the other kinds of diffraction } and what information can be obtained. His particles are probably 1-4 nm, } with another class at probably 6-12 nm. I read somewhere that low-angle } diffraction is probably useful only on particles {0.4 nm. S.a.d. probably } for larger particles. What would people recommend? And right now I could } use a primer/glossary on CBED, SAD, HRED, etc. }
I just finished editing a topical issue of Microscopy Research & Technique, so I'm not biased or anything, but see Vol. 46 #2 & 3 for articles on these subjects. It sounds like the articles by Cowley on nanodiffraction and by Holmstad et al. and by Zuo on CBED will be especially useful. These issues are scheduled to be out
this month.
Disclaimer: Although I edited the journal issues mentioned above, I don't actually get paid more if more get sold. I, therefore, have no monitary interest, only honor and glory. Yours, Bill Tivol
When "cooking" formaldehyde, do so only in a fume hood. Sara Miller
On Thu, 8 Jul 1999 GANTZ-at-med-biophd.bu.edu-at-sparc5.microscopy.com wrote:
} Date: Thu, 08 Jul 1999 12:18:46 -0400 (EDT) } From: GANTZ-at-med-biophd.bu.edu-at-sparc5.microscopy.com } To: MICROSCOPY-at-sparc5.microscopy.com } Subject: Cooking Paraformaldehyde } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Rick: } Yes, I think it is OK to heat paraformaldehyde above 60 degrees. } We have used 1% paraform in combination with 2% glutaraldehyde for } perfusion fix. The source of our recipe is fuzzy. Try Karnovsky, M.J., } 1965. J. Cell Biol. 29:137A. Our recipe is as follows: } } 1) 1 gram PF powder in 50ml DD H20. } 2) Stir and heat to 70 degrees C } 3) Transfer to stirrer (cool) } 4) Add 2 drops 1N NaOH by pasteur pipette } 5) Solution should clear. } 6) Filter thru #1 filter paper. } } This gets diluted by half when combined 1:1 with 4% glut. } Don Gantz } Boston Univ Med School } gantz-at-med-biophd.bu.edu } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Once again I am grateful to the many microscopists offering suggestions to bail me out of my most recent hole. I am posting this summary for all who are interested. I thought Richard Eastwood's and Phil Swab's similar suggestions to use silane derivatives to promote adhesion to the resin was very intriguing. I can't believe I have never heard of that before. I am trying the simplier idea of opening the seeds up before embedding but will probably try a silane derivative as soon as I can scrounge some up. Tom
THE ORIGINAL PROBLEM:
I am having some trouble getting thin sections of some seeds I want to do EM immunolabeling on. I have ttried embedding some seeds in LR White (osmicated) or LR Gold (unosmicated). I took a week slowly infiltrating them with a dilution series of plastic resin but when I went to section them, they just popped out like stainless steel bb's. There is no hint at all that the embedding medium penetrated into them. Any experts out there with hints on fixing seeds?
THE REPLIES:
How intact do you need the seed to be? Since it's already hard and dry, why not whack it with a hammer, then embed the pieces, without the hard seed coat.
Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 Address for courier deliveries: 6319 Pheasant Lane #A-12 Voice: (608) 833-2885 Fax: (608) 836-1969 (please make sure my name is on any fax) oshel-at-terracom.net __________________________________________________________________________
I've used a procedure covered by VA Lindley in Microscopy Research & Techniques 21:355-360 (1992) "A New Procedure for Handling Impervious Biological Specimens" with great success after I struck a similar problem with weevils (never had such a problem with Spurrs resin completely failing to stick to a sample). The paper recommends a pre treatment with gamma-glycidoxypropyl trimethoxysilane (available form Sigma etc) and subsequent embedding with LR White. Its simple & worked amazingly well with re-embedded weevils (they were irreplaceable samples so I stripped off the Spurrs). The paper mentions several samples which they tested the method on, including Salicornia oil seeds. I like it.
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
I saw your email about the seed work. I'd like to suggest you to microwave them during the process of infiltration. For my work, I have microwaved tobacco seeds (twice, 5 seconds each time in a regular microwave) at the beginning of each infiltration step with LR White resin. It worked well for both sectioning and immunogold labeling. Regards. Xinshun (Ding, X.S." {xsding-at-noble.org) __________________________________________________________________________
For the past 16 years I've used an adhesion promotion step to embed difficult materials for diamond knife cross-sectioning. Typically hard, non porous materials such as glass, diamond, silicon, minerals, ceramics, semiconductors, metals, optical coatings and insect parts.
Prepare a 1% solution of Dow Corning's silane co-polymer Z-6040 (3-glycidoxy-propyltrimethoxy silane) in water and alcohol (50/50). Treat your samples in the solution for approximately an hour. Transfer them to filter paper to remove liquid and embed as normal in Spurrs or LR White. These coupling agents are the adhesion promoters used to bind fibers to polymer in fiberglass. Dow has a variety of other silane coupling agents with specificities for epoxies, acrylics, polyesters, phenolics, urethanes, etc. These are typically sold in 55 gal drums but smaller quantities are available.
Phil Swab Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com (Phil Swab) __________________________________________________________________________
The Tips & Tricks site has a few "difficult embedding" discussions archived. Go to:
http://www.biotech.ufl.edu/~emcl/index.html
follow the tips link and either run a search or manually pick through the TEM section. I am a year behind in archiving and will forward anything else I have to you but not to the list as a whole. If there is interest I will post the rest to the archive.
GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks " __________________________________________________________________________
I had a project embedding lettuce seeds and we ran into a similar problem. The best thing we found was to scratch the surface of the seed to break the outter coat. Don't have to cut into it, just a small abrasion on the surface, then put it in a vacuum. I don't have a vacuum oven, so I place them in a desicator and pulled a vacuum with a hand pump until they sank. This made a huge difference for us.
Karen Vaughn Senior Electron Microscopy Technician ICBR EM Core Labatory 214 Bartram Hall Gainesville Fl 32610 phone: 392-392-1184 fax: 392-846-0251 From: "Karen Vaughn " {klv-at-biotech.ufl.edu
Dear Tom, We had similar problems with pollen grains. The trouble was with the difference between the hardness of the pollen and that of the resin. By varying the composition of the resin, we were able to make a sufficient match so that sections could be cut--in our case, thick sections--with many--but not all-- of the pollen grains still in the sections. Good luck. Yours, Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax) --============_-1280767762==_ma============ Content-Type: text/enriched; charset="us-ascii" Once again I am grateful to the many microscopists offering suggestions to bail me out of my most recent hole. I am posting this summary for all who are interested. I thought Richard Eastwood's and Phil Swab's similar suggestions to use silane derivatives to promote adhesion to the resin was very intriguing. I can't believe I have never heard of that before. I am trying the simplier idea of opening the seeds up before embedding but will probably try a silane derivative as soon as I can scrounge some up. Tom
THE ORIGINAL PROBLEM:
I am having some trouble getting thin sections of some seeds I want to do EM immunolabeling on. I have ttried embedding some seeds in LR White (osmicated) or LR Gold (unosmicated). I took a week slowly infiltrating them with a dilution series of plastic resin but when I went to section them, they just popped out like stainless steel bb's. There is no hint at all that the embedding medium penetrated into them. Any experts out there with hints on fixing seeds?
THE REPLIES:
How intact do you need the seed to be? Since it's already hard and dry, why not whack it with a hammer, then embed the pieces, without the hard seed coat.
Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 Address for courier deliveries: 6319 Pheasant Lane #A-12 Voice: (608) 833-2885 Fax: (608) 836-1969 (please make sure my name is on any fax) oshel-at-terracom.net __________________________________________________________________________
I've used a procedure covered by VA Lindley in Microscopy Research & Techniques 21:355-360 (1992) "A New Procedure for Handling Impervious Biological Specimens" with great success after I struck a similar problem with weevils (never had such a problem with Spurrs resin completely failing to stick to a sample). The paper recommends a pre treatment with gamma-glycidoxypropyl trimethoxysilane (available form Sigma etc) and subsequent embedding with LR White. Its simple & worked amazingly well with re-embedded weevils (they were irreplaceable samples so I stripped off the Spurrs). The paper mentions several samples which they tested the method on, including Salicornia oil seeds. I like it.
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
I saw your email about the seed work. I'd like to suggest you to microwave them during the process of infiltration. For my work, I have microwaved tobacco seeds (twice, 5 seconds each time in a regular microwave) at the beginning of each infiltration step with LR White resin. It worked well for both sectioning and immunogold labeling. Regards. Xinshun (Ding, X.S." { {xsding-at-noble.org) __________________________________________________________________________
For the past 16 years I've used an adhesion promotion step to embed difficult materials for diamond knife cross-sectioning. Typically hard, non porous materials such as glass, diamond, silicon, minerals, ceramics, semiconductors, metals, optical coatings and insect parts.
Prepare a 1% solution of Dow Corning's silane co-polymer Z-6040 (3-glycidoxy-propyltrimethoxy silane) in water and alcohol (50/50). Treat your samples in the solution for approximately an hour. Transfer them to filter paper to remove liquid and embed as normal in Spurrs or LR White. These coupling agents are the adhesion promoters used to bind fibers to polymer in fiberglass. Dow has a variety of other silane coupling agents with specificities for epoxies, acrylics, polyesters, phenolics, urethanes, etc. These are typically sold in 55 gal drums but smaller quantities are available.
Phil Swab Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com (Phil Swab) __________________________________________________________________________
The Tips & Tricks site has a few "difficult embedding" discussions archived. Go to:
http://www.biotech.ufl.edu/~emcl/index.html
follow the tips link and either run a search or manually pick through the TEM section. I am a year behind in archiving and will forward anything else I have to you but not to the list as a whole. If there is interest I will post the rest to the archive.
GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks " __________________________________________________________________________
I had a project embedding lettuce seeds and we ran into a similar problem. The best thing we found was to scratch the surface of the seed to break the outter coat. Don't have to cut into it, just a small abrasion on the surface, then put it in a vacuum. I don't have a vacuum oven, so I place them in a desicator and pulled a vacuum with a hand pump until they sank. This made a huge difference for us.
Karen Vaughn Senior Electron Microscopy Technician ICBR EM Core Labatory 214 Bartram Hall Gainesville Fl 32610 phone: 392-392-1184 fax: 392-846-0251 From: "Karen Vaughn " { {klv-at-biotech.ufl.edu
Dear Tom, We had similar problems with pollen grains. The trouble was with the difference between the hardness of the pollen and that of the resin. By varying the composition of the resin, we were able to make a sufficient match so that sections could be cut--in our case, thick sections--with many--but not all-- of the pollen grains still in the sections. Good luck. Yours,
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax) --============_-1280767762==_ma============-- } Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
} When "cooking" formaldehyde, do so only in a fume hood. } Sara Miller
Speaking of formaldehyde I am looking for a formaldehyde free fixitive. I have developed vary serious asthma late in life and I am very sensitive to formaldehyde.
So far Lugol's iodine looks the best for me but I would like something more lasting.
Is formaldehyde the only thing that will fix tissue or just the cheapest. I know it will crosslink almost anything it touches including my sinuses and bronchial tubes. Fortunately it doesn't make it much further than that so it is not a real threat to lungs unless you get in very high concentrations.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
Y'all might be interested to know that "The Usborne complete book of the microscope" by Kirsteen Rogers has been voted (by schoolchildren) as the winner of this year's Rhone-Poulenc junior science book prize.
(information from the recent "Chemistry in Britain")
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
We have a problem with the cooling water on a JEOL 1200EX TEM, and hope that someone can suggest a cause.
The instrument was run for years on a central closed-circuit water chiller which was also feeding several other instruments. About three years ago the chiller was replaced with a new central unit, and it was then the problem started. Over a period of about three weeks the water turns a dark brown to such an extent that it impossible to see through the sight glasses on the flow meters. The brown deposit has been analysed on an EDAX system, and has been shown to be rust. This is further borne out by the fact that the deposit can easily be removed from the flow meters using phosphoric acid.
Thinking that the problem was tied up with the new central chiller, we have now purchased a new chiller unit solely to cool the JEOL. We are using tap water in the system as recommended by the chiller manufacturer, and no additives are being used. The water circuit external to the TEM contains only copper, brass and plastic, and yet the brown rust deposit is still forming just as rapidly as before. Obviously we are very worried that there is some iron component in the TEM itself which is corroding away, and we could be faced with an expensive repair. We would be very interested to hear if anyone else has experienced this problem with a 1200EX.
Regards to all. Bob Phillips ****************** MicroServiS, Huntingdon, Cambs. UK *******************
Although I cannot directly help you with your diffraction questions, I can provide some information about the energy-filtering capabilities of the LEO 912 EFTEM you recently acquired.
I have been using the exact same microscope on organic materials for many years and I must say that you have a powerful scope in your possession! Indeed, I recently published a feature article highlighting what can be done in investigating organic and hybrid materials using this microscope in various modes. The article is going to appear in August:
Energy-filtering TEM of polymers - benefit and limitations of the method Alexander Du Chesne Macromolecular Chemistry and Physics Vol. 200 August 1999 pp. 1813-1830
I have a new task of quantifying the over all fluorescence signal comparing immunolabelled human skin sections from normal vs affected biopsies. Primary antibody was against Laminin 5 with a Texas Red secondary. Images were captured at the same exposure time, normalized and saved as 8bit tiff. Since the region of signal is different for each image, I took the total grey values of the signal, divided by the area(in pixels) of signal and got the average brightness per pixel, which I thought would then be normalized to the region og interest. However, this value doesn't seem to give an indication of how much more or less protien exists in each sample. The average pixel value is only 1.2x different, whereas the difference in area of signal under the histograms is 9x different. But the area under the histogram doesn't normalize for the difference in area of signal, which in this case is the dermal-epidermal juction. There may be more dermal epidermal junction in one image over the other.
What is the proper way to show the real difference in signal? Any suggestions are appreciated and thank you in advance.
} } } Hello, } We need to reduce effects of acoustic interference with high resolution } EFTEM work. Would anyone be willing to share experience with effectiveness } of various wall panels, carpets, or floor tiles for this purpose? } Marek.
this was a big issue in our FESEM installation (LEO982); FETEM should be about the same i imagine.
we experimented with acoustic matting (foam matts) inside the sheet metal boxes of the instrument. the area most critical seemed to be around the ion pump and middle of the column. it worked well enough to get resolution in the 10Ang. range at 30kv. without it the resolution was much poorer.
of course the best way to block the acoustic noise is to eliminate the noise producer. we had to remote install our water chiller...this helped tremendously too.
Brian McIntyre mailto:mcintyre-at-optics.rochester.edu Sr. Engineer lab: 716-275-3058/4875 River Campus EMLab fax: 716-244-4936 University of Rochester Rochester, NY 14620
"The most important thing a father can do for his children is to love their mother." - Unknown
There are some fixatives we use in a microwave where formaldehyde is not a used because of the fumes created after heating. The new fixatives are formulations of glyoxal and they are available from Anatech. The name is Prefer. You have to teach your pathologists to look at tissues fixed in something other than formaldehyde, which may be the difficult part.
Anatech Ltd. 1020 Harts Lake Road Battle Creek, MI 49015 800-262-8324 Good Luck Evanne Maher Zeiss Microm ______________________________ Reply Separator _________________________________
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} When "cooking" formaldehyde, do so only in a fume hood. } Sara Miller
Speaking of formaldehyde I am looking for a formaldehyde free fixitive. I have developed vary serious asthma late in life and I am very sensitive to formaldehyde.
So far Lugol's iodine looks the best for me but I would like something more lasting.
Is formaldehyde the only thing that will fix tissue or just the cheapest. I know it will crosslink almost anything it touches including my sinuses and bronchial tubes. Fortunately it doesn't make it much further than that so it is not a real threat to lungs unless you get in very high concentrations.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
id {01JDCVHA6BOW8WW115-at-denver.du.edu} for Microscopy-at-Sparc5.Microscopy.Com; Fri, 9 Jul 1999 10:26:42 MDT Received: from odin.cair.du.edu (odin.cair.du.edu [130.253.1.2]) by denver.du.edu (PMDF V5.2-29 #28064) with ESMTP id {01JDCVH9IIX88WW0ZH-at-denver.du.edu} ; Fri, 09 Jul 1999 10:26:41 -0600 (MDT) Received: from localhost by du.edu (PMDF V5.2-32 #35157) with SMTP id {0FEM00E0130H4S-at-du.edu} ; Fri, 09 Jul 1999 10:26:41 -0600 (MDT)
On Thu, 1 Jul 1999, ERIC wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Just a quick question to the wealth of knowledge on the microscopy list } server, } } How important is the WPE number on the Eponate 12 Resin bottle from Ted } Pella when calculating the mixture of epon? i.e. should I recalculate the } mixture every time the Eponate 12 Resin has a different WPE number??? } } Any help would be appreciated... } } Eric A. Rosen } Dept. Pathology } UCLA Medical Center } Electron Microscopy Lab... } } Hi,
The WPE numbers from Pella should not change much. A slight change cannot be picked up in the quality of the final block. Simply measure accurately. WPE #s used to be important only when there were huge variations from batch to batch.
The Karnovsky and Ito reference was published only in abstract, which is why you can't find it with conventional searches. The reference is:
Ito S, Karnovsky MJ, 1968. Formaldehyde-glutaraldehyde fixatives containing tri nitro compounds. J Cell Biol, 39:168A-169A (Abstract).
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ On Tue, 6 Jul 1999 rlvaughn-at-UNMC.EDU-at-sparc5.microscopy.com wrote:
} I have lent and lost a reference to a fixative by Karnovsky and (Ito?) . The } paper involved using several different tri-nito compounds in conjunction with } paraformaldehyde and glutaraldehyde. The one the authors liked best used picric } acid with Karnovsky's fixative. I thought it was in Nature? I have ran } searches in medline with no luck. Does anyone remember this fixative and the } original reference? We use it in our Path EM Lab and the CAP inspectors demand } that we have references for everything we do. ( they are driving us nuts) } Thanks in advance. } } Rick Vaughn } } rlvaughn-at-unmc.edu
although I have not seen the issues of the magazine below, I am sure Bill is right. Cowley is always a good reference for diffraction. In addition you may want to check your library for books on electron diffraction by Cowley. May be perhaps a bit theoretical, but worth checking into. If High Resolution is the issue, you may want to check out books by John Spence from ASU. Another source may be the book by L. Reimer about TEM (little plug for my old boss ;-).
If you can't find them let me know. I can probably dig up the exact references for you.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: William Tivol[SMTP:TIVOL-at-WADSWORTH.ORG] } Sent: Thursday, July 08, 1999 3:17:15 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Electron diffraction } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Tina,
Tina Carvalho wrote:
This is the Biological EM Facility, but, with the recent acquisition of a
} LEO 912 EFTEM, I suddenly find non-biologists at my door! There promises } to be some exciting projects.
You will also have some exciting biological projects--that's a terrific machine for getting the maximum information from each electron.
} } One researcher is trying to visualize and characterize Silicon } nanoparticles. They may or may not be crystalline, and they may or may not } form crystals and/or amorphous blobs. The immediate question (for a grant } proposal) is what do we need to do electron diffraction? We can do } s.a.d., but I'm completely unclear as to the other kinds of diffraction } and what information can be obtained. His particles are probably 1-4 nm, } with another class at probably 6-12 nm. I read somewhere that low-angle } diffraction is probably useful only on particles {0.4 nm. S.a.d. probably } for larger particles. What would people recommend? And right now I could } use a primer/glossary on CBED, SAD, HRED, etc. }
I just finished editing a topical issue of Microscopy Research & Technique, so I'm not biased or anything, but see Vol. 46 #2 & 3 for articles on these subjects. It sounds like the articles by Cowley on nanodiffraction and by Holmstad et al. and by Zuo on CBED will be especially useful. These issues are scheduled to be out
this month.
Disclaimer: Although I edited the journal issues mentioned above, I don't actually get paid more if more get sold. I, therefore, have no monitary interest, only honor and glory. Yours, Bill Tivol
Two very important things to remember when comparing digital imaging to film is that you don't have to print every digital image. And you don't have to print every image on high quality (high cost) output media. You do have to develop/process every piece of film. That's how you get real cost savings using digital imaging over film.
Short term: If you are taking a few hundred images per year stick with film. Long term: If you are taking a few thousand images per year move to digital imaging.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
I've had a request here to find out some general information about some of the high voltage EM facilities. I seem to recall there is/was a million volt TEM in Boulder Colorado, are there others? Who would be the contact persons for these facilities? Thanks for your help.
Doug .................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
We are trying to locate Snap-25 on synapses fixed with paraformaldehye, processed for TEM, and poststained with Au. We have achieved very good success with preservation of morphology, but Au deposition has not been adequate (or absent). We have capabilities of embedding in LR Gold at minus 20 deg C., etc. Has anyone done this? Can anyone think of any ideas to encourage the unveiling of this antigen at the synaptic site? Your kind replies might save the day (or our salaries) as they have before.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 readers, I have a problem with very pale staining of my embed 812 } resin thin sections. I often need to use formvar and I cut 50nm thin } sections of cultured cells and fish embryos. I have tried numerous recipes } for both UA (including methanolic) and Lead (Sato's, lead nitrate, lead } citrate). It seems like the stain works well for a couple of times, then } flakes out. I have tried making the lead up fresh each time but even that } fails. The tissue is microwave processed including an en bloc uranyl } acetate step. I take great pains not to get any precipitate by using an } NaOH rinse. I stain 10-15 mins in UA and 5 min. in lead. Does any one have } any suggestions, ideas? What is the best lead stain?? Thanks in advance. } } JoAnn Buchanan } Molecular and Cellular Physiology } Stanford University Scholl of Medicine } Stanford, CA 94305 } } } } Hi,
You have several basic problems. Your sections are too thin. There simply is not enough material present to take up stain. And then they may be on formvar! This compounds the problem doubly, because formvar reduces contrast and blocks stain from one of the sides of the grid. Next, well made citrate (Reynolds) lasts at least a year assuming it is kept correctly. Washing a section with sodium hydroxide after it has been stained with citrate will dissolve the lead citrate off the structures. Do not use it! Use good quality water. Well made lead citrate stain does not precipitate easily. Call me if you have questions and can't get an answer fast. (303-871-3026)
I had sent this email to Geoff earlier and thought we had moved to the email realm. I am really not keen on starting a shootout between the "analogs" and the "digitals". If you want to discuss this further with me, please send me email rather than using the listserver.
Thanks. Michael.
Hey Geoff,
let's not get into a shouting match here. What is cheaper depends a lot on the individual situation and that this is my opinion, it hink I made clear. Just some numbers:
A complete image acquisition and processing system (from us) including camera,TEM adapter, PC, software with on-line shading correction, on-line FFT, etc., but no printer, starts at a little over 43K. And you don't need an extra room for that (if you want to calculate TCO (total cost of ownership) you need to take that into account as well). Photographic quality prints on a Codonics dye-sub printer (you need at least an eyelupe at 16x to start to see some pixelation) are about 50c (that does not include the cost for the printer).
Now, let's assume you shoot one frame of negatives (50) a day. That's about 10,000 negatives a year. I think it is obvious, that it takes just a couple of years to amortize a digital system. And don't forget, you see the images immediately. So for many applications there are advantages that cannot be measured in $, like being able to distribute images seconds after acquisition. Likewise there are other digital advantages (EFTEM), that provide tools not available in non-digital form.
Are you going to the M&M meeting? If so, please stop by at our booth (839). I'd like to show you some stuff you can do with digital imaging.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Geoff McAuliffe[SMTP:MCAULIFF-at-UMDNJ.EDU] } Sent: Friday, July 09, 1999 10:54:21 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: digital archiving/costs } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Doug Cromey,
There are only two HVEMs in the US devoted to bilogical research; the 1 mev in Boulder, CO and 1.2 mev in Albany, New York. There are three others; 2 at Berkley, CA and 1 at Argonne National Labs devoted primarily to Materials research. The Albany HVEM is part of the NIH supported Bilogical Microscopy and Image and Reconstruction Resource (BMIRR) and can be utilized by contacting Dr. Conly Rieder at rieder-at-newton.wadswprth.org (518) 474-6774 Carmen Manella at carmen-at-newton.wadsworth.org (518) 474-2642 or Karolyn Buttle at buttle-at-newton.wadsworth.org (518) 474-6646. For more information about our HVEM or BMIRR check out www.wadsworth.org/spider_doc/bmirr/. The Boulder HVEM is also available and I think Daivd Mastronarde or J. Richard McIntosh would be the contact.
Dave
Dave Barnard HVEM operator Wadsworth Ctr NY State Dept Health Albany, NY 12201 barnard-at-newton.wadsworth.org
I known that this subject come to the surface from time to time, but since I am new here... :) I have being doing the tripod polishing for a couple of months. Now I want to perfect the quality of the sample I prepare. One question is about the best glue for the polishing. I am using a superglue from Faucet Queens and I experimented some problem when polishing III-V semiconductors for XTEM. There is some brand that you can recommend? Thanks for your attention.
Kazuo
PS. If there is a FAQ about this subject, let me know. :)
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
You can use the compressor air to rush the cooling pipe many times until the rust go a way. JEOL EM designer make special connector for it , you can look for the JEOL's instruction.
Doug: Info on the 1.5MeV TEM at Berkeley is available at http://ncem.lbl.gov/frames/hvem.htm The National Center for Electron Microscopy is a U.S. Department of Energy user facility providing scientific researchers with essential resources for electron beam microcharacterization. (http://ncem.lbl.gov/). -Mike O'Keefe
Doug Cromey {doug-cromey-at-ns.arizona.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} the high voltage EM facilities. I seem to recall there is/was a million } volt TEM in Boulder Colorado, are there others? Who would be the contact } persons for these facilities? Thanks for your help. } } Doug } .................................................................... } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : } : Research Specialist, Principal University of Arizona : } : (office: AHSC 4212A) P.O. Box 245044 : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : } : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): } :...................................................................: } http://www.pharmacy.arizona.edu/exp_path.html } Home of: "Microscopy and Imaging Resources on the WWW" } } }
Kazuo, Having used the Tripod Polisher for a number of years, the supergule you are using should be okay. Anyhing cyanocrylate based should be fine.
What is the exact your problem with this glue? It may be specimen based. I have polished Si, AlN-InN-GaN-on-sapphire and tried GaAs. GaAs is a real pain to polish because I found that below 10 microns, the thin edges start to cleave along the charge neutral {110} planes. For the unsuspecting it is really frustrating. Unless you can tripod polish within a very narrow direction I am afraid T Polishing III-V's are always going to be a problem.
I hope this helps, Jonathan ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
The problem we noticed happens specially with III-V materials. We used to polish Si without major problems, but when we started to polish GaAs we started to see small black dots on XTEM. So, when we moved to a different material, GaSb/GaAs we could see that these dots increase in size (some of them half micron in size). Since we do some ion milling to reach the right thickness, these dots act like masks preventing the remove of the material by ion milling. We suspect that these dots are from the glue, specially because they are amorphous.
Our proceedure for the polishing is glue the samples with a piece of Si using Gatan epoxy (face to face), cut and polish both side using sandpaper and diamond lapping film (from 3 to 0.5 um) and finishing with the SBT colloidal blue stuff on a cloth pad. Every step done in a slow rotating polisher. We can't see another source for this contamination, except the glue, but we tried to remove the glue soaking in acetone for 6 hs and rinse with alcohol. The last sample we prepared using 6 hs in acetone resulted in the same problem. Didi you experimented something like this? Thanks for your attention.
Regards,
Kazuo
On Sat, 10 Jul 1999, Jonathan Barnard wrote:
} Kazuo, } Having used the Tripod Polisher for a number of years, the supergule you } are using should be okay. Anyhing cyanocrylate based should be fine. } } What is the exact your problem with this glue? It may be specimen based. I } have polished Si, AlN-InN-GaN-on-sapphire and tried GaAs. } GaAs is a real pain to polish because I found that below 10 microns, the } thin edges start to cleave along the charge neutral {110} planes. For the } unsuspecting it is really frustrating. Unless you can tripod polish within } a very narrow direction I am afraid T Polishing III-V's are always going to } be a problem. } } I hope this helps, } Jonathan } } ******************************************************** } Dr Jonathan Barnard } Analytical Materials Physics } The Angstrom Laboratory, Uppsala University } P O Box 534, SE-751 21 Uppsala, Sweden } Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 } ********************************************************
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
Hello Bob - These JEOLs do not have parts that will rust. I would not worry about some residual rust in the system, it does no harm except to offend your aesthetics. Changing water a couple of times will diminish the colouration but improve nothing. If with clean filters the flow is impeded, I suggest that you use a small pump. Your closed system's could do, for re-circulating a few litres of about 5% hot acetic acid (vinegar). Within an hour it'll clean out any calcium deposits and remaining rust. A few copper ions will go also into solution, but nothing to worry about. It's the only practical way I know to restore good flow to an old EM . Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Sunday, July 09, 2000 9:10 PM, Bob Phillips [SMTP:microservis-at-dial.pipex. com] wrote: } } } Hello All, } } We have a problem with the cooling water on a JEOL 1200EX TEM, and hope that } someone can suggest a cause. } } The instrument was run for years on a central closed-circuit water chiller } which was also feeding several other instruments. About three years ago the } chiller was replaced with a new central unit, and it was then the problem } started. Over a period of about three weeks the water turns a dark brown to } such an extent that it impossible to see through the sight glasses on the } flow meters. The brown deposit has been analysed on an EDAX system, and has } been shown to be rust. This is further borne out by the fact that the } deposit can easily be removed from the flow meters using phosphoric acid. } } Thinking that the problem was tied up with the new central chiller, we have } now purchased a new chiller unit solely to cool the JEOL. We are using tap } water in the system as recommended by the chiller manufacturer, } and no additives are being used. The water circuit external to the TEM } contains only copper, brass and plastic, and yet the brown rust deposit is } still forming just as rapidly as before. Obviously we are very worried that } there is some iron component in the TEM itself which is corroding away, and } we could be faced with an expensive repair. We would be very interested to } hear if anyone else has experienced this problem with a 1200EX. } } Regards to all. } Bob Phillips } ****************** } MicroServiS, } Huntingdon, } Cambs. UK } ******************* } } }
One of the more popular exercises in "Microscopic Explorations" (MSA's middle school microscopy manual) involves looking at sand from around the world. It's interesting, and it leads easily to followup lessons in geology and geography. Joe Neilly, a MICRO volunteer from the Midwest local society, has volunteered to manage MSA's sandpile, and he'll be bringing samples to Portland for use in the Project MICRO workshop at the meeting (Tuesday, 2-5pm). If you have samples from exotic locations (or manufacturing processes, or whatever) will you please bring them (in a film can or ziplock baggie, labelled with location) to the MICRO display in the MSA booth? Ypu'll have the opportunity to make a set of samples on filecard "slides" to use in your own outreach program.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I am looking for the following Nikon Parts, if anyone has or knows where to locate please let me know.
A condenser rack and pinion assembly with condenser holder for a Nikon S-Ke.
A condenser for a Nikon S-Ke
A field lens for a Nikon S-Ke
A lamp housing with lamp socket and the adapter to vertical tube for the Nikon Differential Interference Contract (DIC) Attachment, Model "R" (model "R" is for reflecting illumination).
Hi there Joseph, the s is the old black one right? Yea the DIC is francion yammaoto rather than standard after nomarski is the fine focus on yours coaxial or separate from the rough focus? Ed Sharpe archivist SMECC
Hi, I have been asked by a colleague to ask the list if any one has a reference and/or recipe for Pannett and Comptons saline solution.I think it is to be used with chick embryos and is probably a very old recipe. Thanks in advance, Christine.
I'm having trouble following your logic on digital vs film costs. Original digital images must be very large files if they are to approach the resolution that is routine in film images. Storing thousands of these very large digital image files seems much more of a pain to me than storing a comparable number of film negatives (which is what I archive) in a simple file cabinet. I suspect that this film archiving is less expensive than archiving very large computer files.
Don't get me wrong. I'm completely converted to digital imaging, and spend much of my time working in Photoshop. However, I choose to take the original electron micrographs on film, and to store (archive) them as film negatives (which I can study on a viewing screen). The small percentage of the EMs that are actually used for publications or other serious purposes are scanned with a Leafscan 45 scanner, yielding a digital image file that has as good or better resolution than an original digital image would have had.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
On Fri, 9 Jul 1999, Scott D. Davilla wrote:
} Two very important things to remember when comparing digital } imaging to film is that you don't have to print every digital image. And } you don't have to print every image on high quality (high cost) output } media. You do have to develop/process every piece of film. That's how you } get real cost savings using digital imaging over film. } } Short term: If you are taking a few hundred images per year stick } with film. } Long term: If you are taking a few thousand images per year move to } digital imaging. } } Scott } ----------------------------------------------------------------------- } Scott D. Davilla Phone: 919 489-1757 (tel) } 4pi Analysis, Inc. Fax: 919 489-1487 (fax) } 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com } Durham, North Carolina 27707-2534 web: http://www.4pi.com } }
The latest product people seem to be using is Loctite 460. However, over=
the years many different superglues have been used. The one feature that=
seems most important is that it is fresh. If you let the glue sit around=
it loses it's effectiveness - at least as for as Tripod Polishing goes. =
Basically, you just need to be sure that it is a cyanoacrylate. They wil= l all differ a bit in viscosity and you will need to try a few to find the best one for your application. I am not at my office right now so I don'= t have access to the information, but I have a list of a other glues that have been used over the years. If you have an interest, please contact m= e and I'll put together the list for you on Monday.
You may also want to take a look at our web site as we are offering a Workshop on Tripod Polishing in September. You can take a look at our we= b site at www.southbaytech.com under workshops for details.
Finally, we also have a lot of technical papers on Tripod Polishing which= I would be pleased to send to you - perhaps that along with some of our man= y years of experience manufacturing and using the Tripod Polisher=AE will s= erve as the FAQ list you were looking for. Please contact me off-line if you would like any of this additional information.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Carlos Kazuo Inoki
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Hi,
I known that this subject come to the surface from time to time, but since I am new here... :) I have being doing the tripod polishing for=
a couple of months. Now I want to perfect the quality of the sample I prepare. One question is about the best glue for the polishing. I am usin= g a superglue from Faucet Queens and I experimented some problem when polishing III-V semiconductors for XTEM. There is some brand that you can recommend? Thanks for your attention.
Kazuo
PS. If there is a FAQ about this subject, let me know. :) { {
} Date: Mon, 12 Jul 1999 08:55:00 -0800 } To:University-at-icarus.dur.ac.uk, of-at-icarus.dur.ac.uk, } Durham-at-durham.ac.uk } From:kennedy-at-nsi.edu (Grace Kennedy) } Subject:Re: saline recipe } } Christine, the reference would be: Pannett C.A. & Compton A. (1924) The } cultivation of tissues in saline embryonic juice. Lancet 1924 i, 381-384. } The recipe is, in grams/liter: } } solution A: } NaCl 12.11 } KCl 1.55 } CaCl2 0.77 } glucose 1.0 } } solution B: } Na2HPO4.12H2O 55ml of M/69 solution } NaH2PO4 5ml of M/69 solution } } "Add 4m A to 6 ml B and make up to 100 ml with distilled water. This } saline is too dilute for chicken tissues (Howard, 1953)." } } Howard E. (1953) Some effects fo NaCl concentration on the development of } early chick blastoderms in culture. J.Cell.Comp.Physiol.41, 237-260 } } Our lab is involved in chick/quail chimera studies and we have been using } Hank's balanced salt solution (commercially available) quite successfully. } } The reference above and recipe both came from: } } Lockwood A.P.M. (1961) "Ringer" solutions and some notes on the } physiological basis of their ionic composition. Comp.Biochem.Physiol., } 1961, Vol.2, 241-289 } } Good luck Grace
The Department of Pathology at the SUNY Health Science Center at Syracuse is seeking candidates to perform scanning electron microscopy and light microscopy analysis of environmental media samples, collect and prepare samples, manage data and analyze results. Bachelors degree in environmental science, environmental chemistry or related field or equivalent combination of education and experience required. Electron microscopy preferred.
Informal inquires can be made to this address, but should be accompanied if serious by a cover letter and resume to: Department of Human Resources Dept. D, Job #8680 SUNY Health Science Center 750 East Adams Street Syracuse, NY 13210
I think the confusion comes from the fact, that film indeed has a very high information density, but in most cases you don't need it or not all of it everywhere. Let's look at 2 examples:
1) A low res example: many people are mostly interested in what you can see on the negative with the naked eye. These TEM users mostly print out the entire negative and use these prints for publication, etc. If the microscope is well adjusted, such a negative has a resolution much better than that visible to the naked eye, but most of this resolution does not contain information or only non-relevant information. The idea then is to capture the important information digitally and produce a print from that. For that task, a camera with 1280x1024 will suffice, give or take a few pixels. So for most of these people, the information visible to the eye on a negative can be stored in a single (or perhaps a few) digital images that are 1 MB (2 MB for 12 bit images) in size.
1) A high-res example: Researchers doing high-res TEM are usually not overly interested in what is visible on the negative to the naked eye. Instead, they blow up the negative many times to see the atomic structure (or representation of such). These people look for the highest resolution possible. In most cases, however, the information is not available on the entire negative. For example you may be looking for an interface which is located to a few atomic spacings. 99% of the negative shows areas that are of no interest. On many of my own high-res images (lattice imaging) the negative is only correctly exposed in the center. Areas further out show artifacts due to the changing imaging conditions. For these people, the resolution of the negative is important, but most of the area of the negative is "wasted". Again, a few digital images can provide the same information as the entire negative.
So, if your work falls into one of the two categories, you can replace a negative with a couple of digital images of a few MB size. Of these you can get on the order of hundreds on a single CD at a cost of roughly $1 (for the medium). And image acquisition software will let you search and query a database to find the images you are looking for. In addition, you can annotate you negatives directly on the screen, never have to worry about calibration, send the files over the internet, etc. If not, you are using the wrong software.
The point that Scott wanted to make is, that if you put 50 negatives in the TEM camera, you must develop all 50, costing you on the order of $50 even before you can look at them and decide if they are good. Since you don't want to prepare another sample, you probably take more negatives that you really need. With digital imaging, the cost of acquisition and storing is lower, once you have a system in place. And since you see the image directly, you don't have to store images where the stage shifted or that are overexposed.
Using an off-line camera for digitization is a good solution, as long as you don't need the advantages and possibilities an on-line camera offers. I assume, that you scan the images at a very high resolution for printing purposes. I am not sure, what exactly the printing resolution is that magazines use, but much of the resolution you are trying to get with the Leafscan may be sacrificed again by the printing process. I have used images from 1Kx1K cameras in publications and never had a problem with them.
I am NOT saying, that everybody's work falls into these 2 categories. But from talking to a lot of TEM users I find these uses of a TEM very wide spread. I also think, I should add this disclaimer to the post:
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: A. Kent Christensen[SMTP:AKC-at-UMICH.EDU] } Sent: Monday, July 12, 1999 8:53:05 AM } To: Scott D. Davilla } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: digital archiving/costs } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm having trouble following your logic on digital vs film costs. Original digital images must be very large files if they are to approach the resolution that is routine in film images. Storing thousands of these very large digital image files seems much more of a pain to me than storing a comparable number of film negatives (which is what I archive) in a simple file cabinet. I suspect that this film archiving is less expensive than archiving very large computer files.
Don't get me wrong. I'm completely converted to digital imaging, and spend much of my time working in Photoshop. However, I choose to take the original electron micrographs on film, and to store (archive) them as film negatives (which I can study on a viewing screen). The small percentage of the EMs that are actually used for publications or other serious purposes are scanned with a Leafscan 45 scanner, yielding a digital image file that has as good or better resolution than an original digital image would have had.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
On Fri, 9 Jul 1999, Scott D. Davilla wrote:
} Two very important things to remember when comparing digital } imaging to film is that you don't have to print every digital image. And } you don't have to print every image on high quality (high cost) output } media. You do have to develop/process every piece of film. That's how you } get real cost savings using digital imaging over film. } } Short term: If you are taking a few hundred images per year stick } with film. } Long term: If you are taking a few thousand images per year move to } digital imaging. } } Scott } ----------------------------------------------------------------------- } Scott D. Davilla Phone: 919 489-1757 (tel) } 4pi Analysis, Inc. Fax: 919 489-1487 (fax) } 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com } Durham, North Carolina 27707-2534 web: http://www.4pi.com } }
I wanted to thank all of those who responded to my posting concerning pale staining. It turned out my uranyl acetate was more than 10 years old and had lost it's staining properties. I tried a newer batch as suggested and voila! This whole time I thought it was bad lead stain. It wasn't the formvar or the thinness of the section either. So check the date on those uranyl acetate bottles. Now I have to figure out how to get rid of the old stuff.
Dear listers, our SEM Cambridge S100 needs two integrated circuits (AD7520 & LM308) set on the optic plate and the assistance is not able to give us new circuits. Could someone help us in looking for these two circuits?
Thanks, thanks and thanks
dr Enrico de Lillo Istituto di Entomologia agraria - Universit=E0 Bari - Italy tel. +39 080 5443105 fax +39 080 5442876 email: delillo-at-agr.uniba.it http://193.204.185.103/de_lillo.htm
Postdoctoral Research Associate Advanced Analytical Electron Microscopy of Heterogeneous Catalysts
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
A postdoctoral research associate position is currently available in the Interface Physics Group at the University of Illinois at Chicago (UIC) to develop advanced analytical electron microscopy techniques for characterizing supported catalysts. The successful candidate is expected to apply atomic resolution PEELS, sub-nanometer resolution XEDS, and Z-contrast imaging techniques to the study of active components in supported catalysts. The research associate will work closely with a major industrial company in areas of catalyst development and characterization.
Research in the Interface Physics Group at UIC focuses on the use of atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale. This new catalysis program will focus on the complex interface and surface properties of supported catalysts and small particles. The experimental facilities to perform this research consist primarily of a JEOL 2010 Field-Emission STEM/TEM featuring a 1.4=C5 probe size, "drift free" stage, high-angle annular dark-field detector (Z-contrast), Gatan Imaging Filter, and Noran EDS; a JEOL 3010 conventional TEM with digital imaging capabilities and EDS; and a JEOL 6320 Field-Emission SEM with EDS.
The successful candidate will possess a Ph.D. degree in physics, chemistry, or materials science with a strong background in electron microscopy-based research, and should have demonstrated expertise in atomic resolution STEM and related analytical techniques (PEELS, EDS, etc.). Prior knowledge in supported catalysts or small particles is beneficial. The successful candidate is expected to be highly motivated with an ambition to be part of a developing program for understanding, controlling, and designing heterogeneous catalysts for industrial applications. Excellent interpersonal and written communication skills are necessary. This position is initially for two years and will start as soon as possible. Salary is commensurate with experience. Please send a resume and publication list to Professor Nigel D. Browning at the address below.
UIC is an equal opportunity employer.
Nigel D. Browning, PhD Associate Professor University of Illinois at Chicago Department of Physics (M/C 273) 845 West Taylor Street Chicago. IL 60607-7059. USA Tel: 312-413-8164 Fax: 312-996-9016 http://interface.phy.uic.edu
Nigel D. Browning, PhD Associate Professor University of Illinois at Chicago Department of Physics (M/C 273) 845 West Taylor Street Chicago. IL 60607-7059. USA
I am hoping someone may be able to help me. I am trying to identify mouse macrophages, lymphocytes, eosinophils, neutrophils etc by both scanning and transmission EM. Does anyone have the details of any textbooks which aid in the identification of these types of cells? Thank you in advance,
Kate Scarff CSL, Melbourne Australia e-mail: kate_scarff-at-cslbio.com.au
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
{smaller} As part of a DOE funded program, a postdoctoral position is available for the investigation of the structure-property relationships at internal interfaces in ceramic and ceramic composite systems. The aim of this program is to correlate the experimental results from atomic resolution imaging and microanalysis to produce structural models which can be compared with theoretical simulations. Of particular interest is the use of atomic resolution EELS to characterize the 3-dimensional structure of the interface and quantify the number of vacancies and dopant atoms present in the structure as a function of temperature.=20
Research in the Interface Physics Group focuses on the use atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale. Current research programs involve ceramics, high-Tc superconductors, catalysts, and optoelectronic/high-power semiconducting materials and devices. The experimental facilities to perform this research are comprehensive: a JEOL 2010 Field-Emission STEM/TEM featuring a 1.4=C5 probe size, "drift free" stage, high-angle annular dark-field detector (Z-contrast), Gatan Imaging Filter, Noran EDS, Gatan liquid nitrogen cooling stage, and Gatan heating stage; a VG HB501A Field-Emission dedicated STEM with EDS, EELS and Auger spectrometers; a JEOL 3010 conventional TEM with digital imaging capabilities and EDS; a JEOL 6320 Field-Emission SEM with EDS; and a JEOL JXA733 microprobe. In addition to the electron microscopes, specimen preparation facilities include a Gatan Duo-mill, Fischione precision ion-mill, SouthBay plasma cleaner and Leica Ultramicrotome.=20 The Interface Physics Group has a Silicon Graphics R10000 Power Indigo workstation with the Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The physics department has additional workstations and access to the UIC Convex Exemplar Supercomputer and the National Center for Supercomputing Applications at UIUC. =20
Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevant materials issues and an ambition to be part of a developing program pushing at the frontiers of interface physics. Please send a resume and publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. =20 However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Salary is commensurate with experience. UIC is an equal opportunity employer.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have been following the discussion of digital vs film imaging and archiving with interest and finally decided to put in my two cents. You can argue over which media will be around in 50 years or which is more cost effective. However, the bottom line is to determine which gives you the highest quality data for the intended use.
I am all in favor of digital imaging for routine documentation in that I find users tend to take more images when they are not paying for film. This helps minimize the sampling error that results from drawing conclusions from limited data. Digital imaging is fast, has ecological advantages, and serves very adequately to provide images where speed is important as in diagnostic situations or to facilitate ongoing investigations. In many instances the digital images may be adequate for publication provided minimum enlargement is required.
However, I challenge anyone to show that a digital image printed out on the printers available to most of us (i.e. under $15,000...I use a Codonics 1660) is equivalent to a good darkroom print from a negative in overall sharpness and clarity of image without requiring that the digital image undergo extensive filtering. Adjusting gamma levels, brightness and contrast is done with both production methods as is dogging and burning, but edge sharpening and similar filtering is limited to digital images.
In order to prove the ultimate superiority of film over digital images to myself as well as to my students, I did some careful testing using our SEM. I compared digital images taken at a series of dwell times and resolution settings at magnification of 5000x. Specimen was a calibration grid with latex beads in order to have a specimen ranging in conductivity and density. The specimen was prepared by JEOL and used by their service engineers to check SEM calibration. I also took a Polaroid image of the identical area of the specimen under identical conditions. Polaroid film is not great film but is the most common film used for SEM. I then blew up a portion of the digital images to 18000x (3.6 times) and 36000x (7.2 times). The comparison of resolution vs dwell time drove home the message of the importance of collecting the images at appropriate settings if you intend to enlarge images but at some point, collecting at the highest resolution setting may not yield any additional information but will result in longer collection times and much bigger files.
Then I scanned the polaroid negative into the computer at 300dpi (I routinely use 600dpi for scanning most TEM and SEM negatives). The resultant digital image was compared against the original digital images taken at the higher resolution and longer dwell times and the image from the scanned film was still superior in sharpness with less distortion caused by pixelization. Had I printed the negative in the darkroom and compared the print to the output of the digital files, the difference would have been even more pronounced.
I should note that I encourage users to take images, particularily SEM images, at magnifications that will not require supstantial enlargemnt. However, this has to be balanced against the problems if magifications are selected that result in "empty magnification" with resulting loss of information.
My conclusion is that there is an appropriate use for digital images in the microscopy laboratory. However, if you REALLY need or want to produce the sharpest images with the greatest detail, stick to film. Digital imaging is improving all the time but isn't quite equal to film yet. Don't worry about the storage or cost because the important thing is the data....poor data is a bad buy at any cost.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
I think the confusion comes from the fact, that film indeed has a very high information density, but in most cases you don't need it or not all of it everywhere. Let's look at 2 examples:
1) A low res example: many people are mostly interested in what you can see on the negative with the naked eye. These TEM users mostly print out the entire negative and use these prints for publication, etc. If the microscope is well adjusted, such a negative has a resolution much better than that visible to the naked eye, but most of this resolution does not contain information or only non-relevant information. The idea then is to capture the important information digitally and produce a print from that. For that task, a camera with 1280x1024 will suffice, give or take a few pixels. So for most of these people, the information visible to the eye on a negative can be stored in a single (or perhaps a few) digital images that are 1 MB (2 MB for 12 bit images) in size.
1) A high-res example: Researchers doing high-res TEM are usually not overly interested in what is visible on the negative to the naked eye. Instead, they blow up the negative many times to see the atomic structure (or representation of such). These people look for the highest resolution possible. In most cases, however, the information is not available on the entire negative. For example you may be looking for an interface which is located to a few atomic spacings. 99% of the negative shows areas that are of no interest. On many of my own high-res images (lattice imaging) the negative is only correctly exposed in the center. Areas further out show artifacts due to the changing imaging conditions. For these people, the resolution of the negative is important, but most of the area of the negative is "wasted". Again, a few digital images can provide the same information as the entire negative.
So, if your work falls into one of the two categories, you can replace a negative with a couple of digital images of a few MB size. Of these you can get on the order of hundreds on a single CD at a cost of roughly $1 (for the medium). And image acquisition software will let you search and query a database to find the images you are looking for. In addition, you can annotate you negatives directly on the screen, never have to worry about calibration, send the files over the internet, etc. If not, you are using the wrong software.
The point that Scott wanted to make is, that if you put 50 negatives in the TEM camera, you must develop all 50, costing you on the order of $50 even before you can look at them and decide if they are good. Since you don't want to prepare another sample, you probably take more negatives that you really need. With digital imaging, the cost of acquisition and storing is lower, once you have a system in place. And since you see the image directly, you don't have to store images where the stage shifted or that are overexposed.
Using an off-line camera for digitization is a good solution, as long as you don't need the advantages and possibilities an on-line camera offers. I assume, that you scan the images at a very high resolution for printing purposes. I am not sure, what exactly the printing resolution is that magazines use, but much of the resolution you are trying to get with the Leafscan may be sacrificed again by the printing process. I have used images from 1Kx1K cameras in publications and never had a problem with them.
I am NOT saying, that everybody's work falls into these 2 categories. But from talking to a lot of TEM users I find these uses of a TEM very wide spread. I also think, I should add this disclaimer to the post:
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: A. Kent Christensen[SMTP:AKC-at-UMICH.EDU] } Sent: Monday, July 12, 1999 8:53:05 AM } To: Scott D. Davilla } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: digital archiving/costs } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm having trouble following your logic on digital vs film costs. Original digital images must be very large files if they are to approach the resolution that is routine in film images. Storing thousands of these very large digital image files seems much more of a pain to me than storing a comparable number of film negatives (which is what I archive) in a simple file cabinet. I suspect that this film archiving is less expensive than archiving very large computer files.
Don't get me wrong. I'm completely converted to digital imaging, and spend much of my time working in Photoshop. However, I choose to take the original electron micrographs on film, and to store (archive) them as film negatives (which I can study on a viewing screen). The small percentage of the EMs that are actually used for publications or other serious purposes are scanned with a Leafscan 45 scanner, yielding a digital image file that has as good or better resolution than an original digital image would have had.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
On Fri, 9 Jul 1999, Scott D. Davilla wrote:
} Two very important things to remember when comparing digital } imaging to film is that you don't have to print every digital image. And } you don't have to print every image on high quality (high cost) output } media. You do have to develop/process every piece of film. That's how you } get real cost savings using digital imaging over film. } } Short term: If you are taking a few hundred images per year stick } with film. } Long term: If you are taking a few thousand images per year move to } digital imaging. } } Scott } ----------------------------------------------------------------------- } Scott D. Davilla Phone: 919 489-1757 (tel) } 4pi Analysis, Inc. Fax: 919 489-1487 (fax) } 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com } Durham, North Carolina 27707-2534 web: http://www.4pi.com } }
RFC822 header -----------------------------------
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(InterMail v03.02.07.07 118-134) with SMTP id {19990713163717.KYQW4651-at-oemcomputer} ; Tue, 13 Jul 1999 16:37:17 +0000 Message-Id: {3.0.3.32.19990713124000.00893b70-at-postoffice.worldnet.att.net} X-Sender: edhirsch-at-postoffice.worldnet.att.net X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.3 (32)
Kazuo,
I would like to tell you that I represent Allied High Tech Products and we are a manufacturer of equipment for sample preparation and a distributor of laboratory supplies for sample preparation.
The superglues that have been discussed should work well and be completely acetone soluble. We have also found certain waxes to work very well when applied in small amounts. The wax is completely acetone soluble and will clean from the sample easily and all waxes are not the same.
Could the black dots be from the SiC grinding steps? As you know the III-V materials are very brittle and extra care should be taken when preparing them. The sandpaper you speak about might be causing the difficulties too. I would eliminate it and use a larger size of diamond lapping film for grinding such as 15 micron and then 6 micron continuing with 3,1 and 0.5 micron.
We also have a tool, the MultiPrep, that is a semi-automatic tool for preparing SEM and TEM samples. The MultiPrep is excellent for providing reproducible results and reliable results eliminating variations that can occur with hand tools. A detailed brochure is available on our website at http://www.alliedhightech.com/pdf/multiprep.pdf please note this is a pdf file and Adobe Acrobat reader is required to view it.
If I may provide you with additional information please let me know offline.
Sincerely, Ed Hirsch
************************************************* Edward A. Hirsch Product Application Specialist Allied High Tech Products 2376 East Pacifica Place Rancho Dominguez, CA 90220 ph: (919) 846-9628 vm:(800)675-1118 x245 fx: (310)762-6808 http://www.alliedhightech.com *************************************************
I would like to tell you that I represent Allied High Tech Products and we are a manufacturer of equipment for sample preparation and a distributor of laboratory supplies for sample preparation.
The superglues that have been discussed should work well and be completely acetone soluble. We have also found certain waxes to work very well when applied in small amounts. The wax is completely acetone soluble and will clean from the sample easily and all waxes are not the same.
Could the black dots be from the SiC grinding steps? As you know the III-V materials are very brittle and extra care should be taken when preparing them. The sandpaper you speak about might be causing the difficulties too. I would eliminate it and use a larger size of diamond lapping film for grinding such as 15 micron and then 6 micron continuing with 3,1 and 0.5 micron.
We also have a tool, the MultiPrep, that is a semi-automatic tool for preparing SEM and TEM samples. The MultiPrep is excellent for providing reproducible results and reliable results eliminating variations that can occur with hand tools. A detailed brochure is available on our website at http://www.alliedhightech.com/pdf/multiprep.pdf please note this is a pdf file and Adobe Acrobat reader is required to view it.
If I may provide you with additional information please let me know offline.
Sincerely, Ed Hirsch
************************************************* Edward A. Hirsch Product Application Specialist Allied High Tech Products 2376 East Pacifica Place Rancho Dominguez, CA 90220 ph: (919) 846-9628 vm:(800)675-1118 x245 fx: (310)762-6808 http://www.alliedhightech.com *************************************************
I had some success with tripod polishing of GaAs and thought that I would ramble on about how I did it. BTW, I highly recommend that GaAs is NOT the material that you use to learn how to tripod polish when you are teaching yourself how to do it. What I found when I went back to doing GaAs with the TP after having some experience with the technique with other materials was that I had to protect the edges of the GaAs sample especially when using the lowest grit sizes. I originally did this by trying to "encapsulate" the edges by smearing the sides, surface and back with a coating of epoxy (Epo-Tek 353ND). This worked somewhat if the coating is thin. If it is too thick, the epoxy pulled away from the sample leaving a small gap. Thinner was better. After Ron Anderson presented his technique of embedding a small sample in a hollowed-out dimple in Si, I borrowed his idea and tried coring a desired area of GaAs out with a small ultrasonic drill that I made with the thin walled stainless steel tubing available from Small Parts, Inc. I then drilled a matching hole in a piece of Si such that when I put the plug into the Si and filled it with epoxy, the epoxy "wall" around the sample was thin.
I believe that the epoxy helps prevent the edges of the GaAs from chipping out while using the lowest grit sizes and rescratching the polished surface of the GaAs. When the sample gets too thin, this scratching breaks the sample. The Si was used to gauge the thickness of the sample by color changes. (See John McCaffrey's paper on this subject.)
Sorry, I don't have the sizes for the tubes that I tried. I tried a whole (or is it "hole") assortment of different sizes because the ultrasonic core drill doesn't quite give plugs or holes of exactly the same diameter as the inner and outer diameters of the tubes. I silver soldered the tubes into stainless steel parts that I had made that would fit the ultrasonic drill. This presented a rash of problems too, but with the appropriate jigs, this could probably be done more easily than I did it. Because of these issues, the smallest plug of GaAs that I used was about 1.5 mm. It was really nice having the Si to help with determining the thickness of the sample.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I apologize in advance for this message, but it is sort of an emergency approach to covering up a mistake. A few days ago someone sent me a message asking for an address where my book 'Vacuum Methods in Electron Microscopy' could be purchased in India. I mistakenly deleted the message before I could send out a reply, and so don't know who sent it nor the address for the reply. Therefore, I'm putting this general message on the listserver in the hopes that it will reach the proper person. If it does, I'd appreciate a reply from him/her.
Probably the best place to order the book in India is from: Affiliated East-West Press Pvt Ltd G-1/16 Ansari Road New Delhi 110 002, India Tel: 11 327 9113 or 11 326 4180 Fax: 11 326 0538
However, the book can also be ordered directly from the publisher, Portland Press of London, through their web site, which also gives addresses of distributors throughout the world: http://www.portlandpress.co.uk
While we're on the subject, information about the book (outline, reviewers' comments, etc.) can also be found at the SPI web site: http://www.2spi.com/catalog/books/book48.html and it can be purchased from SPI.
Another apology, W. C. Bigelow
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Hi Ya'll: Several month's ago, I asked the listserver community for help/suggestions on the installation of a drive (other than the present Syquest upgrade that is available from Kevex) to replace the 44Mb Bournoulli's. I would like to know if anyone has upgraded their system without the Kevex upgrade. If someone hasn't upgraded their system, but understands how to do it, I would surely appreciate your help. In my first post, several people responded directly to me, and now I have found that I have accidentally deleted their responses. If you would like to respond again, that would be fantastic. Thanks in advance for your help. Sorry for any inconvenience.
Regards, Michael Coviello Lab Manager MRCEDM UT Arlington
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 6 - October 14, 1999
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2050 (Includes room and board, text, handouts, supplies)
Application Deadline: August 3, 1999
Admission application and information: Carol Hamel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, polarized light, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy, multiphoton excitation fluorescence microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratiometric-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual research problems with the faculty.
I am in charge of organizing a section on teaching microscopy for a local (SF bay area) meeting this October. If you are interested in participating let me know. In addition to some presentations, I think we can do posters. If you would like to display a poster, I would be happy to put it up for you if travel to the meeting is not possible.
The meeting is to be held Sat. Oct. 2, 1999 at San Francisco State University and is a joint meeting of the Northern California Society for Microscopy, an MSA local affiliate, and the Cal State University EM lab group. Abstracts will be published in Microscopy Research and Technique.
Abstracts are due Aug. 2, but I will lobby for an extension if you have something good and can't get it in until after MSA. You can check a web page about the meeting at:
http://online.sfsu.edu/~camicro/
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I feel somehow compelled to answer all these postings, while on the other hand I think, we are using too much bandwidth for this already. But it's hard to judge without getting feedback. Should we continue this? Nestor, is it OK to continue?
Debby Sherman wrote:
} However, I challenge anyone to show that a digital image printed out } on the printers available to most of us (i.e. under $15,000...I use a } Codonics 1660) is equivalent to a good darkroom print from a negative in } overall sharpness and clarity of image without requiring that the digital image } undergo extensive filtering. Adjusting gamma levels, brightness and } contrast is done with both production methods as is dogging and burning, but } edge sharpening and similar filtering is limited to digital images.
There is a whole chain of software and hardware that has to work hand = in hand to provide good results. And unfortunately the weakest link = usually determines the final quality. That may be the printer, the camera or = the software. And that is why it is important to buy complete and tuned systems, not a camera from here and software from there. But let me quote yourself: "However, the bottom line is to determine which gives you the highest quality data for the intended use." So, if an edge filter does give you better results for the intended use, why not use it? An edge filter is just a procedure like dogging or burning, it's just not available for film. One caveat: if digital filters are used blindly, i.e., without knowing what the effects and side-effects are, they can = be dangerous! Nobody wants to introduce artifacts and then interpret them as features. Especially important for using FFT filters!
} The comparison of resolution vs dwell time drove home the message of } the importance of collecting the images at appropriate settings if = you } intend to enlarge images but at some point, collecting at the highest } resolution setting may not yield any additional information but will result in } longer collection times and much bigger files.
I agree completely. In fact, could you send me those images? I'd like = to see them myself. The reason for this behavior is manyfold, but there is really a point where increasing the resolution will not yield more information. It has to do with the digitization of the scanning voltage and transmission noise, but I don't think we have to go into that at this point.
} Then I scanned the polaroid negative into the computer at 300dpi (I } routinely use 600dpi for scanning most TEM and SEM negatives). The } resultant digital image was compared against the original digital images taken } at the higher resolution and longer dwell times and the image from = the } scanned film was still superior in sharpness with less distortion caused by } pixelization. Had I printed the negative in the darkroom and = compared the } print to the output of the digital files, the difference would have been } even more pronounced.
I'd be curious as to how you judged the sharpness. I've had strange results when comparing printers: I printed out the same image on a Codonics 1600 and a Lexmark Inkjet printer for a couple of hundred Dollars. For the inkjet I used the best (glossy) paper, the Codonics = has it's own paper. The images looked sharper and crisper on the Ink-jet! However, when I took an eye lupe to look at the image in detail, there was no question. While the Codonics hardly showed any pixelation even = at a mag of 16x, the ink jet showed nothing but a few colored dots. I concluded from that, that either my eyes tricked me (Am I getting old??), or that the inkjet process with the dithering produces an image that the brain perceives as "sharper". I'd be interested in hearing how you compared the images.=20 Also, If I understood correctly what you did, you recorded the image on Polaroid, then digitized it on a scanner with 300 DPI. Then you = compared it to an image acquired directly from the SEM. I am not sure, what the size is of a Polaroid negative, but if it's 4x5, you should compare = that to a digital image of 1500x1200 pixels taken with a dwell time as to = get the same exposure time as the negative. If you don't, you are comparing apples to oranges. For example, if you compare that to a digital image of much higher resolution, you probably have to drop the dwell time and thereby increase the noise. And if you then look at the image at a higher mag, you may perceive that as a decreased "sharpness" (or increased "fuzzyness").
} Don'tworry about the storage or cost because the important thing is the } data....poor data is a bad buy at any cost.
At 07:37 AM 7/13/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Who really cares about 50 year old data? Archeologists might but what was printed 50 years ago may be referenced. The challenge is to obtain the highest quality images now for immediate use and publication. If one does publish, the standard printed medium runs at 133 lpi. Since digital capture can achieve much higher levels, the difference is in tonal range. This is an ongoing battle between digital photography and film photography. The range of film is pure analog, while that of digital is binned in nature. But with high resolution (10-bits or more per pixel), digital does a good job.
Suppose that you have a film master. What now? If you make a print and send it to a magazine, they will flatbed or drum scan it for master output. The resolution is still 133 lpi. And the D range is less than the original neg.
Therefore, the mechanics are such that having a film original is not the end. One must convert it to digital for printing. Starting with digital in the first place eliminates that phase. And it eliminates any loss incurred by the conversion process. What is archived is an original. With film, the archive is an approximation, or at best, a decent copy.
I don't dismiss digital all that much. In fact, I dismiss film more than digital.
The Department of Geology and Geophysics at the University of Calgary invites applications from qualified persons interested in joining our technical support staff as an Electron Microprobe & Petrology Technician.
The Department is in the process of purchasing a new electron microprobe, expected to be in place in early 2000, and requires a senior technician to support the new instrument and related activities. The position is full-time and permanent. Compensation includes a competitive salary and generous benefit package.
A full description with application procedures is available from the Department of Geology and Geophysics web page: http://www.geo.ucalgary.ca/
please be so kind and do not be upset for crossposting.
I have just start section RECIPES at www.coleoptera.org in directory {Other useful things} at bottom of the section { for collectors}
Please be so kind and send to me any recipes of for relaxing specimens, microscopy, mounting of speciemns etc., glue recipes, how to get mould off the specimen, anything waht you feel that could be handy or interesting for entomologists. Thank you very much for your response
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptiste (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
http://www.coleoptera.org Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.37 / Virus Database: 30 - Release Date: 6.4.1999
I've just been given the OK to buy a microwave oven for our EM facility. I would like to hear the good and/or bad on the Electron Microscopy Sciences' EMS-820 Precision Pulsed Laboratory Microwave Oven verses the Ted Pella' Pelco 3450 Laboratory Microwave System.
TIA for any information that would be useful in trying to make a decision on which one to buy.
George Lawton Chief Electron Microscopist Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75235-9039 Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
Hi, I'm now to the list... Anyone out ther familiar with a Unitron series N with a 35mm camera attachment? I seem to have one sitting on my coffee table, and would like to try to take some pictures with it. How do I get light down to the camera....where is the shutter on the microscope? Whats the sidelamp supposed to do? Whats a xenon lamp? The overhead lamp seems to be working, and I can look at stuff pretty darn close...but I want pictures. Also, anyone know how I adjust the eyepeices in a bit? I know it must sound stupid, but I've got a small brain, hence, a small head. Thanks for any help. -Matt B
PS For what it worth, the whole thing is about 2.5 feet tall and weighs in at about 90lbs give or take 10.
_________________________________________________________ Do You Yahoo!? Get your free -at-yahoo.com address at http://mail.yahoo.com
I would think any good e.m. histology text would do. I use: Johannes A. G. Rhodin (1974) Histology A Text and Atlas Pub Oxford University Press Library of Congress Cat No. 73-90374
It's good for TEM of mammalian cells and tissue (a lot of pictures are of rat or mouse) but has no SEM. I would have thought that TEM would be of more use in identification because you can see size and shape of nucleus as well as bodies and organelles in cytoplasm. It's usually a fairly straight forward process to identify most blood cells by TEM unless you're just looking at bits.
Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: Kate Scarff To: Microscopy
Gary and List - A long but interesting discussion, mostly factual. However, some conclusions may be wishful thinking, with the arguments woven into a system of beliefs. Lets talk TEM images - SEM images are 'designed' for digital all the way. Electrons forming the image are not digital expressions -unless we are prepared to use billions of bits. The image intrinsically is best recorded in analogue. Practical arguments like printer's resolution comes later. TEM resolution is identical at high and low magnifications (to a point). So a medium power image, which still has a tremendous depths of focus, can be enlarged 20 and more times, revealing detail not apparent at lower enlargement; this simple technique side-steps the very difficult task of focussing at say 200 000x. Only at about 30x enlargement (difficult to achieve with an enlarger), insufficient electrons form the image and it becomes "noisy". In practice, it makes sense to take an image at say 40k and then enlarge to 600k and beyond. Try that with a CCD in the scope! Alternatively it means that the entire negative should be scanned at no less than 900ppi for proper digital archiving, including good continuous tone rendition. Handy viewing and long term save storage of negs are further arguments in favour of the photo process. Subsequent scanning and or printing for use are another discussion.
I conclude that all other considerations are choices for a particular laboratory or personal preference, based on: required quality, magnification range commonly used, number of images taken and realistic archiving needed etc. Based on such considerations some labs will find digital recording and archiving saves MONEY, which is the common denominator of all factors. Please do not argue on quality - the TEM neg wins every time and will do so for the foreseeable time.
Disclaimer PST supplies TEM films AND digital cameras.
Cheers Jim Darley PS From my system of beliefs on archiving images: Medical laboratories need to keep most images for many years. In a university laboratory I believe that long term central storage is just a huge liability. I would leave the archiving to the user: "please remove your photos within four weeks or they may be placed into the circular file on the floor". ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Wednesday, July 14, 1999 9:17 AM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] wrote:
printing process . . . . . . Therefore, the mechanics are such that having a film original is not the end. } One must convert it to digital for printing. Starting with digital in the } first place } eliminates that phase. And it eliminates any loss incurred by the conversion } process. What is archived is an original. With film, the archive is an } approximation, or at best, a decent copy. } } I don't dismiss digital all that much. In fact, I dismiss film more than } digital. } } Cheers, } Gary Gaugler, Ph.D. }
I have only set up various bernoulli and syquest drives on the 8000/Delta but will speculate...
Two conditions must be satisfied. The hardware interface and formatting. I suspect that any SCSI 1 drives could be made to work (standard hardware interface). DMON will need to be setup appropriately to logically access the drive. After that, the question is if the RT-11 formatting program will function with a different drive. Since I recently acquired an iXRF system, my 8000/Delta is "in the corner" awaiting a decision on its' dispositon - so I can't try much.
Perhaps this vague info is food for thought anyway. (Comments Warren?)
Woody White McDermott Technology Inc. ____________________Reply Separator____________________
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Hi Ya'll: Several month's ago, I asked the listserver community for help/suggestions on the installation of a drive (other than the present Syquest upgrade that is available from Kevex) to replace the 44Mb Bournoulli's. I would like to know if anyone has upgraded their system without the Kevex upgrade. If someone hasn't upgraded their system, but understands how to do it, I would surely appreciate your help. In my first post, several people responded directly to me, and now I have found that I have accidentally deleted their responses. If you would like to respond again, that would be fantastic. Thanks in advance for your help. Sorry for any inconvenience.
Regards, Michael Coviello Lab Manager MRCEDM UT Arlington
by rose.muohio.edu (8.9.1/8.9.1) with ESMTP id JAA85678 for {microscopy-at-MSA.Microscopy.com} ; Wed, 14 Jul 1999 09:10:28 -0400 Message-Id: {199907141310.JAA85678-at-rose.muohio.edu} Received: from CASSERVER1/SpoolDir by casmail.muohio.edu (Mercury 1.44); 14 Jul 99 09:10:29 -5 Received: from SpoolDir by CASSERVER1 (Mercury 1.44); 14 Jul 99 09:10:08 -5 To: microscopy-at-Sparc5.Microscopy.Com
Well, it seems someone has decided that they needed our CPD baskets more = than we did (what for I can't imagine), so now I am trying to replace them. I am part= icularly looking for the larger (18-22mm) diameter screw together baskets made of b= rass and stainless steel - they look like little metal pill cases/boxes. Here's a = good description I borrowed from EMS for the smaller versions:
" This basket is brass with a stainless steel mesh attached to both ends; = the mesh opening is 300=B5m. (The inside diameter is 12.5mm, 15mm height. (16mm ODx= 17mmH). To use, simply screw both halves together..."
} } I conclude that all other considerations are choices for a particular } laboratory or personal preference, based on: required quality, magnification } range commonly used, number of images taken and realistic archiving } needed etc. } Based on such considerations some labs will find digital recording and } archiving saves MONEY, which is the common denominator of all factors. Please } do not argue on quality - the TEM neg wins every time and will do so for the } foreseeable time. } There is one aspect in which digital imaging in a TEM via CCD or imaging plates is superior to film: quantitative measurement of image intensity. I'm surprised no one has mentioned this yet. At least for materials science purposes, comparison of images to simulations is becoming more and more important. In order to do this, a researcher needs a linear quantitative electron intensity measurement device, which is very difficult to achieve with film. Film certainly has a higher information density than digital recording, but in some cases digital methods provide access to information not accessible by film.
-Paul
Paul Voyles voyles-at-research.nj.nec.com voice: (609) 951-2627 fax: (609) 951-2496 NEC Research Institute, 4 Independence Way, Princeton, NJ 08540 USA
We are currently trying to embed Drosophila ovarioles for TEM, to look at cell membranes. Unfortunately we are having a difficult time preserving the membranes. Using the following protocol, our cells completely lacked membranes, the cytoplasm was very dense and full of ribosomes but there were white spaces where membranes should have been.
1. Dissect in PBS 2. Fix in 2% gluteraldehyde, 1.5% paraformaldehyde, 1.5% acrolein in .1 M cacodylate buffer 90 min -at- RT 3. 4.
6. Wash in water 7. Dehydrate through a series of ET-OH washes: 30%, 50%, 70%, 90%, 95%, 2X 100% 8. 2X 10 min each in Propylene Oxide (PO) 9. Infiltrate with Spurr's 10. Polymerize24-48 hrs -at- 65oC. 11. After sectioning, contrast with 2% UA in 50% EtOH, and lead citrate
In a previous experiment, in which ovarioles were dissected in similar fixative but without acrolein and the rest of the protocol was the same, mitochondrial and nuclear membranes were well preserved but the cell plasmalemma (what we are interested in!) was absent or incomplete. I don't think it is any of our reagents because we don't have this problem with any other tissues.
Any advice would be greatly appreciated. Thanks in advance
Michelle
#################################################### Michelle Peiffer Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:mlk101-at-psu.edu ####################################################
} } I've just been given the OK to buy a microwave oven for our } EM facility. I would like to hear the good and/or bad on the } Electron Microscopy Sciences' EMS-820 Precision Pulsed } Laboratory Microwave Oven verses the Ted Pella' Pelco 3450 } Laboratory Microwave System. } } TIA for any information that would be useful in trying to make a } decision on which one to buy. } } George Lawton } Chief Electron Microscopist } Microscopy and Imaging Service Center } UT Southwestern Medical Center at Dallas } 5323 Harry Hines Blvd. } Dallas, Tx 75235-9039 } Phone: 214-648-7291 } eMail: George.Lawton-at-email.swmed.edu
Hi George We bought the Pelco oven.
I went to the microwave workshop at U of Berkeley (Kent Macdonald's lab) last January and I thoroughly recommend it. From that workshop and my own evolving protocols, I recommend that you get the vacuum chamber as well if you are going to process any difficult specimens such as nematodes, drosophila larvae, plant material. Without the chamber, whole drosophila larve are still white grubs with black tips after osmium treatment, (even after 24 hours normal post fixation). With the vaccum chamber, they are black all the way through after osmium treatment in 45 minutes in the microwave. You do need the low power settings - 200 watts to do this. Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
I already replied to Mike directly on the matter, but basically concur with what you wrote. Many of the controllers had a format utility in their firmware that could be used to format the drive. I don't think the stock RT-11 one worked. I think I still have the SCSI format utilities that Kevex provided to work with a couple of the PDP-11 SCSI controllers. They might work in a pinch.
I told Mike that one of the issues is the amount of addressable disk space. The RT-11 DL driver normally only handled two 10-MB partitions, but could be coaxed up to four. The DU: driver can handle up to eight 32-MB partitions. So, you are limited to either 40 MB or 256 MB of total usable space depending on which controller you have. Anything beyond that will just sit idle, unless you got a fancy controller someplace else. But then, 40 MB seemed like a lot of space on those PDP systems.
At 08:21 AM 7/14/1999 -0500, Woody wrote: } I have only set up various bernoulli and syquest drives on the 8000/Delta } but } will speculate... } } Two conditions must be satisfied. The hardware interface and formatting. I } suspect that any SCSI 1 drives could be made to work (standard hardware } interface). DMON will need to be setup appropriately to logically access } the } drive. After that, the question is if the RT-11 formatting program will } function with a different drive. Since I recently acquired an iXRF system, } my } 8000/Delta is "in the corner" awaiting a decision on its' dispositon - so I } can't try much. } } Perhaps this vague info is food for thought anyway. (Comments Warren?) } } Woody White } McDermott Technology Inc.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Matt B } ------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Hi, I'm now to the list... Anyone out ther familiar with a Unitron series N with a 35mm camera attachment? I seem to have one sitting on my coffee table, and would like to try to take some pictures with it. How do I get light down to the camera....where is the shutter on the microscope? Whats the sidelamp supposed to do? Whats a xenon lamp? The overhead lamp seems to be working, and I can look at stuff pretty darn close...but I want pictures. =
Also, anyone know how I adjust the eyepeices in a bit? I know it must sound stupid, but I've got a small brain, hence, a small head. =
Thanks for any help. -Matt B
PS For what it worth, the whole thing is about 2.5 feet tall and weighs in at about 90lbs give or take 10.
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No doubt this topic has been beaten to death in this go-around. I suspect it will arise again (and again). Here's another opinion --modest as usual.
I see no good reason --none-- for using film recording on an SEM given the cost effectiveness, efficiency, flexibility, superior control and recording quality of digital image capture systems. A compelling justification for many labs is eliminating the environmentally suspect chemicals used in processing film. We ignore the noxious chemicals used in producing the silicon, of course. For TEM, the high cost of appropriate-quality CCD recording systems limits their application. Of four TEMs in the lab I use, we've been able to justify the cost of equipping just one (a high-end FEGTEM) with a 1K x 1K CCD system. The preference for digital vs film recording is mixed, depending on application, but digital recording has some considerable advantages. Resolution is not always the overwhelming issue: Some other considerations that favor digital recording over film in TEMs are immediate availability of recorded images, linear intensity response, and superior quantum efficiency. The compromise of where to place the CCD in the TEMs imaging system (above, at , or below the viewing screen, or at the end of an electron spectrometer) has important implications for intended uses. As to image storage, the recordable CD is a suitable medium that can be read by nearly every desktop computer today. I often wish they had more storage capacity and faster recording. Also, most TEM images require some king of processing, and once you've put in the work you'll probably want to store the processed images too.
Larry
Larry Thomas Washington State University Pullman, WA
Does anyone know of a supplier for Hyrax mounting media? This , apparently, is the mounting medium of choice for diatoms.
Thanks, Mike Baxter mykkb-at-juno.com ___________________________________________________________________ Get the Internet just the way you want it. Free software, free e-mail, and free Internet access for a month! Try Juno Web: http://dl.www.juno.com/dynoget/tagj.
I know I've harped on this before, but I thought I'd bring a recent article to everyone's attention. Digital data is not as long lasting as we would like to believe, see this article in a recent Newsweek for more information.
July 12, 1999 Issue of NEWSWEEK MAGAZINE TECHNOLOGY: History: We're Losing It "They told us digital data would last forever. They lied. How do we save the past before it all disappears?"
Previous stories/articles on this topic: Business Week 20 Apr 98 Scientific American, January 1995
Don't get me wrong, I'm not a Luddite, we're going digital too. Caveat Emptor!
Yours, Doug Cromey .................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
Labs lacking in CCD cameras aren't doomed to a non-quantitative existence. The density D on the film is related to electron exposure E by the simple linear equation D = bE + d, where b and d are constants, for appropriate values of Dmax (Zeitler & Bahr, 1962, J. Appl Phys. 33, 847). While CCD cameras and position-sensitive electron counters are terrific, film can still be used to make quantitative measurements without the capital cost of a CCD camera.
Mike Lamvik MCNC, Research Triangle Park, NC ---------------------------------
} There is one aspect in which digital imaging in a TEM via CCD or } imaging plates is superior to film: quantitative measurement of image } intensity. I'm surprised no one has mentioned this yet. At least } for materials science purposes, comparison of images to simulations } is becoming more and more important. In order to do this, a } researcher needs a linear quantitative electron intensity measurement } device, which is very difficult to achieve with film. Film certainly } has a higher information density than digital recording, but in some } cases digital methods provide access to information not accessible by } film. } } } -Paul } } } Paul Voyles voyles-at-research.nj.nec.com } voice: (609) 951-2627 fax: (609) 951-2496 } NEC Research Institute, 4 Independence Way, Princeton, NJ 08540 USA
I am afraid the linearity of film can not be assumed unless for images with very little contrast and very little density. The actual relation between the density D of the film and the current density I is described reasonably well be the equation:
D = Dmax ( 1 - exp(-c I t) )
where Dmax is the maximum density of the film, c the speed of the film, I the electron density and t the exposure time.
Unfortunately, this is not the end of the story. When digitizing the image, another non-linear function gets involved. The pixel value P is
P = a exp( -D ln(10) )
where a is proportional to the illumination intensity usually adjusted such that for an 8-bit image the dynamic range is within the pixel values from 0 to 255.
The expression for I as a function of P (which is what a CCD camera delivers (at least in very good approximation)) is arguably not linear. Should someone decide to correct for the non-linearity, I would like to point out that Dmax is not really a constant. It depends on many factors like the age of the developer, how long film is developed and also on the illumination used to digitize the film. Diffuse illumination usually provides a smaller value for Dmax than parallel illumination.
If this has not discouraged you, please find more details in:
E. Voelkl, F. Lenz, Q. Fu and H. Lichte, "Density Correction of Photographic Material for Further Image Processing in Electron Microscopy", Ultramicroscopy, 55 (1994) 75--89
Best regards, Edgar
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Does anyone know of a supplier for Hyrax mounting media? This , apparently, is the mounting medium of choice for diatoms.
Thanks, Mike Baxter mykkb-at-juno.com ___________________________________________________________________ Get the Internet just the way you want it. Free software, free e-mail, and free Internet access for a month! Try Juno Web: http://dl.www.juno.com/dynoget/tagj.
a colleague is after references and/or info on how to calculate specimen temperature rise due to a focussed electron beam in a 200kV TEM. He has been studying nickel particles (approximately 0.1 - 5 micron in diameter) on holey carbon coated copper grids. Under a focused beam larger particles tend to melt but smaller ones end not to.
He would like to estimate the temperature rise in different diameter particles and for different metals/materials as well. Any help would be appreciated. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
Three post doctoral fellowships are vacant at the Centre for Materials Research, University of Oslo.
For more detailed information please contact professor Tore Amundsen, phone no.. +47 22 95 87 36. e-mail: tore.amundsen-at-fys.uio.no
The positions are funded by the Norwegian Research Council for two years. The projects are divided into the following:
"Thin films produced by ALE-methods". The applicants should be familiar with this method.
"Production of films by sol/gel and nanoparticle methods". The applicants need considerable experience within the field, and show ability to work independently and show intiative.
"Atomic modelling of crystals, surfaces and interfaces". Interest and experience in computersimulation and a background in solid state physics or solid state chemistry is required.
The fellowships will be available from the end of August beginning of September 1999.
Applications must reach the Centre for Materials Research by August 15.= 1999.
Please send your application or contact: Centre for Materials Research University of Oslo Gaustadaalleen 21 0349 Oslo, Norway
************************************************
for Centre for Materials Research, University of Oslo.
Dear All, I am a beginner in SEM/TEM and would appreciate your help. I am labelling leukemia cell cultures for SEM with 10nm immunogold labelled ABs and therefore have to silver enhance the label. Usually this is done after fixing the cells with glutaraldehyde. However I read that post fixation with osmium tetroxide dissolves the silver layer - can I use the silver enhancer after the osmium tetroxide treatment? (The cells will be on polycarbonate filters). Thanks in advance
Claudia Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT12 2EE ++44(0)181 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
} } I try to find the best way in prooving the depth of a burn in rat skin } } (staining, counting hair follicle, antibody). Does anybody has experience } } with that? } } } } Another question is to stain parrafin slides with x-gal. Is there } } anyprotocol out there which works. I tried to replace xylene with } } Clearite3.But unfortuantly I didn't find any blue cells on these slides. } } What is the best protocol for X-Gal in fresh frozen sections. } } } } Lars Steinstrasser } } } } Trauma Burn Research Lab } } 1510 MSRB I } } 1150 West Medical Center Drive } } Ann Arbor, MI 48109-0666 } } } } Tel: (734) 615-2510 } } Fax: (734) 763-7932 } } Pager: (734) 936-6266 #1129 } } } } Private } } 218 Village Green Blvd. #203 } } Ann Arbor /MI 48105 } } Tel: (734) 747-6200 }
_______________________________________________________________ Get Free Email and Do More On The Web. Visit http://www.msn.com
I recently fielded a call from a Father looking for a good, yet inexpensive light microscope for his Daughter. He is looking for a tool in the 5 to 100X range, and something that is "solid", not the junk they sell in toy stores.
If you have any ideas for him please contact Dr. Thomas via E-mail at lathomas-at-aol.com
My feeling is that this might not be easy because of difficulty of estimating conductive heat loss through the carbon, which will depend on how good the contact is.
However, there is a good section in the textbook by Ludwig Reimer "Transmission Electron Microscopy", which describes modeled and experimental approaches and has a number of references which may be useful.
Wharton Sinkler
} } } Hi all, } } a colleague is after references and/or info on how to calculate specimen } temperature rise due to a focussed electron beam in a 200kV TEM. He has } been studying nickel particles (approximately 0.1 - 5 micron in diameter) } on holey carbon coated copper grids. Under a focused beam larger particles } tend to melt but smaller ones end not to. } } He would like to estimate the temperature rise in different diameter } particles and for different metals/materials as well. Any help would be } appreciated. Cheers, } } } Mark Blackford } TEM Group } Materials Division, ANSTO } PMB 1, } Menai, N.S.W. } Australia } 2234 } } Phone 61 2 9717 3027 } Fax 61 2 9543 7179 } } Disclaimer: } The views expressed in this E-mail message do not necessarily represent the } official views of ANSTO from which this message was conveyed. } } } }
When anyone asks for information about buying microscopes etc for schools you should point them to the Project Micro pages which were put together by Caroline Schooley.
I ran across a replacement for both Hyrax and Naphrax at http://www.emsdiasum.com/ems/chemicals/adhesive.html - called MeltMount from Cargille.
Bill Miller
At 02:41 PM 7/14/99 -0400, mykkb-at-juno.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
t 02:41 PM 7/14/99 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Has anyone out there performed a careful examination of stage drift in a high-res SEM? If so, what drift rate have you measured? We recently purchased a new FEG SEM and have been quoted that drift on that SEM should be less than 40 nm over a 6 minute period (under ideal sample and SEM conditions). Since this could amount to a significant drift over the many hour time period required for an OIM run, I would like to find out what drift rate others have measured (or been quoted) for their SEMs for comparison.
A related question for the OIM users is: how do you account for the drift (and/or minimize it) on longer OIM runs to avoid distortion of the OIM map?
TIA, Richard Fonda
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
New this year to Microscopy & Microanalysis is the MSA Booth! Attendees
will conveniently find in the MSA island (# 739): the Technologists Forum, Certification Board, Project MICRO, the Education Committee's Book Display, and the new MSA Membership Desk. The computer workshop will be in booth # 749, adjacent to the MSA island.
Come by and learn about the many activities and services of the society.
Look for the "MSA in the sky" down isle 700! See you in Portland.
Janet H. Woodward - for the Chairs of the MSA Committees
Thank you all for the responses received so far. I don't know why my original posting didn't go through correctly, but here it is again. We are using osmium and en bloc UA, but somehow it got cut out of the original post.
We are currently trying to embed Drosophila ovarioles for TEM, to look at cell membranes. Unfortunately we are having a difficult time preserving the membranes. Using the following protocol, our cells completely lacked membranes, the cytoplasm was very dense and full of ribosomes but there were white spaces where membranes should have been.
1. Dissect in PBS 2. Fix in 2% gluteraldehyde, 1.5% paraformaldehyde, 1.5% acrolein in .1 M cacodylate buffer 90 min -at- RT 3. 4.
6. Wash in water 7. Dehydrate through a series of ET-OH washes: 30%, 50%, 70%, 90%, 95%, 2X 100% 8. 2X 10 min each in Propylene Oxide (PO) 9. Infiltrate with Spurr's 10. Polymerize24-48 hrs -at- 65oC. 11. After sectioning, contrast with 2% UA in 50% EtOH, and lead citrate
Any advice would be greatly appreciated. Thanks in advance
#################################################### Michelle Peiffer Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:mlk101-at-psu.edu ####################################################
} Dear List members: } } I am in the market for a basic microtome, any } recomendations/information would be appreciated. } } Thank you, } Dawn } } Dawn
If you are interested in a refurbished (used) microtome, I carry a number of makes and models. All are completely refurbished to manufacturer's specifications and come with a 90 day warranty. Please contact me for details and price quote.
-- Ford M. Royer, MT(ASCP) Analytical Instruments, Ltd (Refurbished Histology, Cytology, & General Lab Equipment) 9921 13th Ave. N. Minneapolis, MN 55441-5004 800-565-1895 phone 612-929-1895 fax web site: http://www.aibltd.com
Nestor, RHS people etc. I guess there will be similar pages this side of the pond. Any suggestions from different countries?
you wrote: ==== Colleagues
When anyone asks for information about buying microscopes etc for schools you should point them to the Project Micro pages which were put together by Caroline Schooley.
Nestor Your Friendly Neighborhood SysOp ==== Best wishes
Stephan Helfer +44(0)131 248 2865; fax +44(0)131 248 2901 ------------------------------------------- To most people 'solutions' mean finding the answers. But to chemists solutions are things that are still all mixed up. (Science Explained)
Unsubscribe, please ---------------------- Douglas R. Keene Shriners Hospital Microscopy Facility 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
This question on stage drift reminded me of a correction technique that is in John Russ' book, "Imaging Processing Handbook" in chapter 5 on Processing images in frequency space. Basically, you take the Fourier transform of the image and the Fourier transform of a line vector that represents the length of the displacement vector. You divide the first transformed image by the second transformed image and then transform the resulting image back. It is really slick.
This might work on an image that is "summed" over many frames, but unfortunately, I don't think that it will work on an image that is collected sequentially over a long time. I think there, that you would have to apply a shear to the image to get point registry back.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Richard W. Fonda To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Has anyone out there performed a careful examination of stage drift in a high-res SEM? If so, what drift rate have you measured? We recently purchased a new FEG SEM and have been quoted that drift on that SEM should be less than 40 nm over a 6 minute period (under ideal sample and SEM conditions). Since this could amount to a significant drift over the many hour time period required for an OIM run, I would like to find out what drift rate others have measured (or been quoted) for their SEMs for comparison.
A related question for the OIM users is: how do you account for the drift (and/or minimize it) on longer OIM runs to avoid distortion of the OIM map?
TIA, Richard Fonda
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
Responding to the message of {98FF9B038A-at-rbge.org.uk} from "Stephan Helfer" {S.Helfer-at-rbge.org.uk} : } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Nestor, RHS people etc. } I guess there will be similar pages this side of the pond. Any } suggestions from different countries? } Apart from being written in English (American), t.hese pages should be applicable anywhere in the world. If you know of other microscopes that are not mentioned in this list, I am sure that Caroline Schooley would love to get details of them (and a review) from you. She can be found at:
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
__________________ Stuart McKernan stuartm-at-tc.umn.edu Associate Director Office:(612) 626-7594 CIE Characterization Facility, University of Minnesota Desk: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
In a message dated 7/15/99 1:47:22 PM, walck-at-ppg.com writes:
} This question on stage drift reminded me of a correction technique that } is in John Russ' book, "Imaging Processing Handbook" in chapter 5 on Processing } images in frequency space. Basically, you take the Fourier transform of } the image and the Fourier transform of a line vector that represents the length } of the displacement vector. You divide the first transformed image by } the second transformed image and then transform the resulting image back. } It is really slick. } } This might work on an image that is "summed" over many frames, but } unfortunately, I don't think that it will work on an image that is collected } sequentially over a long time. I think there, that you would have to apply } a shear to the image to get point registry back.
FT deconvolution is definitely the wrong approach for this problem. If the stage drift is uniform then as Scott notes the effect on the raster scanned image is a shear in the image. Exactly the same thing can be found in satellite images which are raster scanned as the platform orbits, and the images are again sheared in one direction. If you know the shear (which in the satellite case is easy and in the SEM case might be if you measured the location of a feature before and after a known time interval), then you can stretch the image back pretty easily. You can do it interactively in programs like Photoshop or could write a simple macro to do it in NIH-Image.
Now that I've gotten past the hyperactive spam filter ...
Amenex has a 1976-vintage ETEC Autoprobe SEM with three WDS spectrometers. Most of the instrument still functions acceptably (with the able assistance of Ken Converse) but we have noticed substantial deterioration of the oxygen dot maps in the last few years. The sodium maps come out OK; nitrogen has always been pretty much hopeless; but carbon works OK.
We've been told that our TAP crystal may have deteriorated or become contaminated. What can we do about that ? Are there any adjustments or alignments that could be tweaked so as to pick up the oxygen radiation better ?
Has anyone got a spare TAP crystal for sale or trade ? We have several spares of other crystals ...
Any advice or suggestions would be greatly appreciated.
Best regards, George Langford, Sc.D. amenex-at-amenex.com http://www.amenex.com/
It is not a good idea to perform silver enhancement after osmium tetroxide treatent, because the deposited osmium can also act as a nucleating site for silver deposition, and you will silver enhance the osmicated regions of your sample as well as the gold particles.
The greatest risk of etching of deposited silver is when both osmium tetroxide and uranyl acetate are used. In the absence of uranyl acetate, etching is often not a problem. In addition, Burry and co-workers have found that in situations where it is a problem, it may be greatly reduced by using 0.1 % osmium tetroxide instead of 1 %; this has been found to give similar levels of staining (Burry, R.W. Pre-embedding immunocytochemistry with silver-enhanced small gold particles, p. 217-230. In M. A. Hayat (Ed.). Immunogold silver staining: Principles, methods and applications. CRC Press, Boca Raton (1995)).
Etching may be prevented altogether by gold toning. The procedure for this i= s:
1. After silver enhancement, wash thoroughly with dionized water. 2. 0.05 % gold chloride: 10 minutes at 4=B0C. 3. Wash with deionized water. 4. 0.5 % oxalic acid: 2 mins at room temperature. 5. 1 % sodium thiosulfate (freshly made) for 1 hour. 6. Wash thoroughly with deionized water and embed according to usual procedure.
(Reference: Arai, R., et al, Brain Res. Bull., 1992, 28, 343-345).
I hope this helps.
Rick Powell Nanoprobes, inc.
****************************************************************** * PLEASE NOTE NEW E-MAIL ADDRESS: rpowell-at-lihti.org * * * * NANOPROBES, Incorporated | US Toll-free: (877) 808-2101 * * 25 East Loop Road, Suite 113 | Telephone: (919) 510-0590 * * Stony Brook, NY 11790-3350, | Fax: (919) 510-0590 * * USA | www.nanoprobes.com * ******************************************************************
} a colleague is after references and/or info on how to calculate specimen } temperature rise due to a focussed electron beam in a 200kV TEM. He has } been studying nickel particles (approximately 0.1 - 5 micron in diameter) } on holey carbon coated copper grids. Under a focused beam larger particles } tend to melt but smaller ones end not to. } } He would like to estimate the temperature rise in different diameter } particles and for different metals/materials as well. Any help would be } appreciated. Cheers,
Dear Mark, In order to calculate the heat input, one needs to have the stopping power of the material for electrons of the appropriate energy. For 200 kV e- in Ni,
that would be about 1.9 MeV*cm^2/g (I have the tables for Fe & Cu, so I in- terpolated). Only the collision stopping power contributes to the temperature
rise, since the radiation from brehmsstrahlung is not absorbed in small par- ticles. Perhaps David Joy's Monte Carlo program can be used (or modified) to integrate the heat loss over the volume of a particle to get the total heat
input. The loss would be through radiation (proportional to T^4) and con- duction. The former would depend on the total surface area, and the latter would be a function of contact area (among other variables). The particles can probably be treated as black bodies (at most, a constant would be put into the equation; the particle would then be a gray body). The conductive loss is more difficult to calculate, and it probably dominates. Both the loss
terms are smaller for larger particles--volume-to-surface and volume-to- contact-surface--but if the contact surface can be assumed to be the same fraction of the total surface for particles of all sizes, perhaps a simple model can be used. That assumption is probably good for spheres, bad for plates, and varies for other shapes. If all the particles are spheres, the size of the smallest sphere which melts can be used to determine the parameter for my proposed simple model. Good luck. Yours, Bill Tivol
I have contacted five major U. S. SEM supply vendors and only two of them sell 1/8" pin mount storage boxes that will accommodate high samples (approx. 1/2" to 5/8" clearance). The least expensive and highest box is out of stock for at least two weeks and I need some ASAP. (The cost of these boxes range from 1.35 to 6.00 dollars each!) Any suggestions would be appreciated. You can probably guess who the vendors that I contacted are but I would be interested to find any new players out there. Thank you.
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269-2131 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
I just want to thank to everybody that had send his piece of comment for my problem. The problem was solved just moving to a different superglue. It seems that the brand we were using was just leaving some residues even after full night in acetone. Now we moved for the locitite superglue, as some of you suggested, and the sample is clean of the black dots we observed before. Thanks again.
Regards,
Kazuo
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Joe -
All the information that your friend needs is on the Project MICRO web page, http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html under "How to buy a microscope" and "Supply sources".
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Dear Ph.Ds-to-be: We have a post-doc position available at Emory University, Department of Neurology in Atlanta, GA. Specifically, we are looking for someone with EM experience to participate in research on the neuropathology of Huntington's disease and Fragile X syndrome. If you are interested in this position, or know of anyone who might be, please contact me at the following email address. Thank you very much.
Hong Yi
=========================== Hong Yi EM Supervisor Emory University School of Medicine Department of Neurology 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30322 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
I do not have www, only free mail. Could someone remind me of the Image Analysis List Server's email adress or find it in the archives of this server? Also an SPM- and TEM-vendor type list server if anyone knows of one. Please respond to my email.. Thank you. Jeff Day/ 'JD'
At 02:14 PM 7/15/99 , you wrote: } I have contacted five major U. S. SEM supply vendors and only two of them } sell 1/8" pin mount storage boxes that will accommodate high samples } (approx. 1/2" to 5/8" clearance). } The least expensive and highest box is out of stock for at least two weeks } and I need some ASAP. (The cost of these boxes range from 1.35 to 6.00 } dollars each!) Any suggestions would be appreciated. You can probably guess } who the vendors that I contacted are but I would be interested to find any } new players out there. Thank you. } } James S. Romanow } The University of Connecticut } Physiology and Neurobiology Department } Electron Microscopy Facility } U-131 } Storrs, CT 06269-2131 } bsgphy3-at-uconnvm.uconn.edu } 860 486-2914 voice } 860 486-1936 fax }
Pella makes the 16160 which is a tall storage box for 12 stem mounts. They are $2.25 ea or $15.50/10. I have some extras right now if you need them and Pella is out of stock. I had no trouble getting them.
Am I right in understanding that the "Cargille" liquids for RI determination are organic solvent-like? If so, does anyone know a series of liquids for examining particles of polymers which would be swollen or even dissolved by such liquids?
Thanks in advance,
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
A little help if possible. I am used to working with human cells, tissue sections etc. Recently a member of staff has arrived with two samples of wood !!!, one of pine shavings with pine dust, the other sample is of MDF again dust and shavings. The question she asked was what size and shape are these particles.
The tools that I have to hand are either a confocal microscope with KrAg laser or a zeiss axiovert with a z focus motor, with phase and fluorescent illumination and deconvolution software.
{/color} The way that I thought I might proceed is to find a wood fibre specific stain that would autofluoresce and then collect a z series, deconvolve and render.
Any suggestions as to dyes or basically any other suggestions would be gratefuly received. The end point of this is to try to determine the difference between pine and mdf and to suggest how far these may travel in the lungs before lodging.
Peter
{nofill}
Peter McHardy Technology Services Manager, Beatson Institute, Glasgow University, Garscube Estate, Bearsden, Glasgow G61 1BD Tel 0141 330 4818 Fax 0141 330 4127 http://www.beatson.gla.ac.uk
I have one Microsoft Publisher file, size 99k (probably written in Publisher 97 or higher). There is a data for our SEM. But I can' t read this file with my Publisher version 3.0. Please help me, if someone has a Publisher 97 or 98.
-- Henrik Kaker, Ph.D. SEM-EDS Laboratory Metal Ravne, Koroska cesta 14 Ravne, Slovenia, Phone: +386 602 21 131 Fax: +386 602 20 436 Mailto:Henrik.Kaker-at-guest.arnes.si http://www2.arnes.si/guest/sgszmera1/index.html
You might consider replacing the TAP crystal with, or adding, a layered synthetic crystal. There is a WSi crystal with a 2d spacing of ~ 60 angstroms that works very well for C, N, and O. This crystal will have much higher count rates than the TAP, but will sacrifice some wavelength resolution. One vendor I am aware of is Osmic in Michigan. I don't know if they could fit one to your ETEC.
Jim McGee
{} {} {} {} {} {} {} {} {} {} {} {} {} James J. McGee (jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
TEL: (803) 777-6300 FAX: (803) 777-6610
On Thursday, July 15, 1999 5:13 PM, George Langford, Sc.D. [SMTP:amenex-at-amenex.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 probing Microscopists ! } } Now that I've gotten past the hyperactive spam filter ... } } Amenex has a 1976-vintage ETEC Autoprobe SEM with three } WDS spectrometers. Most of the instrument still functions } acceptably (with the able assistance of Ken Converse) but } we have noticed substantial deterioration of the oxygen } dot maps in the last few years. The sodium maps come out } OK; nitrogen has always been pretty much hopeless; but } carbon works OK. } } We've been told that our TAP crystal may have deteriorated } or become contaminated. What can we do about that ? Are } there any adjustments or alignments that could be tweaked } so as to pick up the oxygen radiation better ? } } Has anyone got a spare TAP crystal for sale or trade ? We } have several spares of other crystals ... } } Any advice or suggestions would be greatly appreciated. } } Best regards, } George Langford, Sc.D. } amenex-at-amenex.com } http://www.amenex.com/
I am taking SEM images, BEI mode of paper samples. I have problems to differenciate between the binders used in paper and the epoxy resin used to embedd the paper samples. I wonder if there is a way of staining the resin in order to get better contrast when taking backscattered images. I know that OsO4 can be used to stain some binders,but I also want to stain the resin.
Johann Gebauer has done an excellent job of rebuilding Philips EM300 specimen tips for us. Just sent him all of your broken parts.
Gebauer Machining 117 Ridgecrest Rd. Ithica, NY 14850 607 273-5049
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269-2131 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
I have embedded reproductive tracts from parasitoid wasps and leps with none of the problems you are experiencing. Maybe I've just had good luck? Anyway, the only things that differ between our protocols are that I dissect in the primary fixative, not in PBS, and the primary fixative is phosphate buffered glutaraldehyde/formaldehyde pH 7.3 without any acrolein. The only components of the repro tract that have proven poorly fixed are more or less mature eggs. Let me know what finally works for you. Good luck!
Eloise--
Eloise L. Styer Veterinary Diagnostic Lab University of Georgia 43 Brighton Rd. PO Box 1349 Tifton, Ga 31793 912 386-3340
Michelle L. Peiffer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Thank you all for the responses received so far. I don't know why my } original posting didn't go through correctly, but here it is again. We are } using osmium and en bloc UA, but somehow it got cut out of the original } post. } } We are currently trying to embed Drosophila ovarioles for TEM, to look at } cell membranes. Unfortunately we are having a difficult time preserving } the membranes. Using the following protocol, our cells completely lacked } membranes, the cytoplasm was very dense and full of ribosomes but there } were white spaces where membranes should have been. } } 1. Dissect in PBS } 2. Fix in 2% gluteraldehyde, 1.5% paraformaldehyde, 1.5% acrolein in .1 M } cacodylate buffer } 90 min -at- RT } 3. } 4. } } 6. Wash in water } 7. Dehydrate through a series of ET-OH washes: 30%, 50%, 70%, 90%, 95%, } 2X 100% } 8. 2X 10 min each in Propylene Oxide (PO) } 9. Infiltrate with Spurr's } 10. Polymerize24-48 hrs -at- 65oC. } 11. After sectioning, contrast with 2% UA in 50% EtOH, and lead citrate } } Any advice would be greatly appreciated. Thanks in advance } } #################################################### } Michelle Peiffer } Electron Microscope Facility for the Life Sciences } The Biotechnology Institute for Research and Education } 1 South Frear Lab } University Park, PA 16802 } 814-865-0212 email:mlk101-at-psu.edu } ####################################################
We have a Sorvall Porter-Blum JB4 Microtome. It seems to be in good working condition. However, it does not have any manual.
If any of you have a manual of this machine, I will really appreciate sending me a copy.
Thanks in advance,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
Robert, the Cargille RI oils that I use do fine for synthetic fibers (nylon, rayon, etc.). You may want to contact McCrone Corp. to see if they have specific RI oils for polymer particles. 800-622-8122. Hope this helps Mike Bucker Consolidated Labs of Va. Richmond, Va. 23219
} } } "Robert H. Olley" {R.H.Olley-at-reading.ac.uk} 07/16 5:32 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Am I right in understanding that the "Cargille" liquids for RI determination are organic solvent-like? If so, does anyone know a series of liquids for examining particles of polymers which would be swollen or even dissolved by such liquids?
Thanks in advance,
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
For the best and most trained of ETEC technicians, the alignment of the WDS crystals is a daunting task. The TAP crystal is not a particularily sensitive crystal as far as cleaning, a simple quick swabing with ethanol should remove most contamination. However, the ETEC crystal mount has four mounting screws that are adjusted to control the tilt of the crystal in two dimensions, the crystal height and alignment with the Rowland circle as well as the curvature of the crystal.
Ken is a very good technician on the ETECs and was my technical backup as a field technician. However, I was the only technician, that I know of, that was trained by the gentleman who set up the WDS systems in the factory. I know from setting up many systems that I would often spend an entire week adjusting a single crystal for proper operation. These are fully focussing Johansson WDS optics that are capable of measurements to one ten thousandth of an Angstrom. However, the adjustment screws are very course and touchy to properly adjust. Added to this is the proper adjustment of the detector tape, a metal tape that both maintains the line of sight of the detector to the crystals as well as maintaining the proper detector alignment to the Rowland circle. The ETEC WDS spectrometer also requires an accurate alignment of the spectrometer housing to the sample chamber.
More than likely, your spectrometer is in need of a full alignment that may well cost you thousands of dollars and a couple of weeks of work or more to accomplish.
I can put you in touch with a gentleman who can provide you with new crystals, contact me if needed, but more than likely you have need of a complete overhaul of the spectrometer system.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
-----Original Message----- } From: George Langford, Sc.D. {amenex-at-amenex.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Robert H.Olley wrote: ================================================= Am I right in understanding that the "Cargille" liquids for RI determination are organic solvent-like? If so, does anyone know a series of liquids for examining particles of polymers which would be swollen or even dissolved by such liquids? ==================================================== The Cargille family of refractive index fluids and immersion oils cover a wide range of liquid chemistries, all are considered broadly to be "organic oils" but they range in composition from pure hydrocarbon to some that are siloxanes (most "laser" liquids) to yet others that are perfluorinated polyethers. There are probably some classes of liquids I have l have not mentioned.
Solvency depends on the polymer you are trying to characterize.
A custom fluid can usually be produced for a very specific refractive index at a specified wavelength within a specified viscosity range. Without knowing the polymer involved, it is not possible to know how the choice of liquids might be further constrained. A fluid that would swell a porous polymer particle might not necessarily dissolve it. Of course there is always the chance that one of the standard fluids would work.
Contact me off line and maybe we can design a specific liquid that will accomplish what you require. More information about the standard line of Cargille fluids can be found on our website given below.
Disclaimer: SPI Supplies offers the full range of Cargille fluid products so we are not a disinterested third party.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
Does any one have any information on the Sakura sapphire knives or where we could find the company?
Thanks. ML 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
You were one of several factory trained technicians: I was one, Ken was another, Hank Bebe was also trained. I was trained by "Tung Tsu" who trained Darrell Jackson. Ask Darrell how many spectrometers he screwed up before being trained by Tung Tsu.
Earl
"Allen R. Sampson" 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 } -----------------------------------------------------------------------. } } For the best and most trained of ETEC technicians, the alignment of the WDS } crystals is a daunting task. The TAP crystal is not a particularily } sensitive crystal as far as cleaning, a simple quick swabing with ethanol } should remove most contamination. However, the ETEC crystal mount has four } mounting screws that are adjusted to control the tilt of the crystal in two } dimensions, the crystal height and alignment with the Rowland circle as well } as the curvature of the crystal. } } Ken is a very good technician on the ETECs and was my technical backup as a } field technician. However, I was the only technician, that I know of, that } was trained by the gentleman who set up the WDS systems in the factory. I } know from setting up many systems that I would often spend an entire week } adjusting a single crystal for proper operation. These are fully focussing } Johansson WDS optics that are capable of measurements to one ten thousandth } of an Angstrom. However, the adjustment screws are very course and touchy } to properly adjust. Added to this is the proper adjustment of the detector } tape, a metal tape that both maintains the line of sight of the detector to } the crystals as well as maintaining the proper detector alignment to the } Rowland circle. The ETEC WDS spectrometer also requires an accurate } alignment of the spectrometer housing to the sample chamber. } } More than likely, your spectrometer is in need of a full alignment that may } well cost you thousands of dollars and a couple of weeks of work or more to } accomplish. } } I can put you in touch with a gentleman who can provide you with new } crystals, contact me if needed, but more than likely you have need of a } complete overhaul of the spectrometer system. } } Allen R. Sampson } Advanced Research Systems } 317 North 4th. Street } St. Charles, IL 60174 } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com } } -----Original Message----- } } From: George Langford, Sc.D. {amenex-at-amenex.com} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } Date: Thursday, July 15, 1999 1:09 PM } Subject: Haaalp ! How can I improve oxygen dot maps in microprobe ? } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 probing Microscopists ! } } } } Now that I've gotten past the hyperactive spam filter ... } } } } Amenex has a 1976-vintage ETEC Autoprobe SEM with three } } WDS spectrometers. Most of the instrument still functions } } acceptably (with the able assistance of Ken Converse) but } } we have noticed substantial deterioration of the oxygen } } dot maps in the last few years. The sodium maps come out } } OK; nitrogen has always been pretty much hopeless; but } } carbon works OK. } } } } We've been told that our TAP crystal may have deteriorated } } or become contaminated. What can we do about that ? Are } } there any adjustments or alignments that could be tweaked } } so as to pick up the oxygen radiation better ? } } } } Has anyone got a spare TAP crystal for sale or trade ? We } } have several spares of other crystals ... } } } } Any advice or suggestions would be greatly appreciated. } } } } Best regards, } } George Langford, Sc.D. } } amenex-at-amenex.com } } http://www.amenex.com/ } } } } } }
I had always worked with TEM and had developed an interest in performance=
monitoring through my service days, hence my RMS book "Maintaining & Monitoring the Transmission Electron Microscope" and work on "Quality" in=
electron microscopy.
When I first started using SEM my instincts naturally took me into testin= g out these machines too. I just had to measured resolution, contamination=
rate and drift rate on every instrument that I used, either as a consulta= nt or within our own courses. Often having the students make the tests.
In relation to contamination and drift rate tests we would work over a 20=
minute period at 50,000X. A level that all instruments were capable of reaching. The test specimen was one of my gold on latex, here there is a=
0.5mm block of dried latex (0.24um), that has been multi coated (sputtere= d 6-10 times at different angles) with gold to create a coarse (latex) and = a fine (gold) structure. As a result the specimen is well earthed!
So we ran the tests and to my surprise the results were almost always as follows - within a 20 minute period the drift was less than the resolutio= n of the instrument i.e. we could not detect the drift. At first I thought=
this was an error but repeated tests over this period on a number of instruments gave the same results. We were looking at a double exposure = to measure the movement of the latex spheres. If any drift was detected subsequent tests put this "image shift" down to charge. Re positioning t= he specimen in the stage and checking the stage earth solving most problems.= =
You learn a good deal about the SEM simply by trying to perform this type=
of test. High voltage stabilization periods for example!
I have not met anyone else who has done this but they must be out there somewhere? Most of the SEM users I worked with in the early days, who ha= d more SEM experience than I, thought to measure drift rate and contaminati= on rate a bit of a joke!
Hope this helps?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Post Doctoral position for recent PhD (no longer than 5 years) or PhD to be.
We have a post-doc position available at the NIH, NIDDK. Specifically, we are looking for someone with a Cell Biology interest to participate in research on intracelllular lipid trafficking. Technically someone with Electron Microscopy experience and cryo-immunogold cytochemistry experience would be ideal. However our laboratory is equipped to train an individual interested in devoting their efforts to this line of research. There are several cytological projects of importance to the laboratory that could be approached with these techniques. If you are interested in this position, please contact me at the following e mail address
Thank you very much.
E.Joan Blanchette-Mackie, PhD
-------------------------------------------------- Dr. Joan Blanchette-Mackie Chief; Section of Lipid Cell Biology NIDDK / NIH Builing 8, Room 427 8, Center Drive Bethesda MD 20892 - 0850
With regard to the discussion on Naphrax™, it along with some of the other "older" mountants, like Canada Balsam and Aroclor™, have been "replaced" with other more "user friendly" alternatives. Some of the original mountants contained PCBs, although I have forgotten just what the less desirable feature was about Naphrax.
Cargille Laboratories has develped a line of replacements, they are part of their line of "Optical Quality Thermoplastics". They are PCB-free. Their RI 1.704 product is the direct replacement for Naphrax. It has an Abbe V dispersion of 24. It is available as a thermoplastic liquid as well as what is called their Quick-Stick™. Both products are described on our website below. These products are also offered by several of the other firms supplying microscopy products to microscope users.
Disclaimer: SPI Supplies is a distributor for the Cargille Laboratories line of products for microscopy.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
Glad to hear it, Earl. I had never heard from either Hank or Ken that they had been properly trained in WDS, and still have not. To the contrary, after I first came on board, received orientation in Danbury for a week and received basic training in Hayward for two weeks, I was teamed with Darrel for a couple of weeks for WDS training and then left to install a dozen instruments through out the midwest because there had been no one available to do the installations for the previous two years or more. Forgive me if I am a little touchy on this subject, but I had to go into many situations where ETEC had simply been unable to completely install instruments that had been delivered years before. I realize that I entered ETEC on the cusp of their acquisition by Perkin-Elmer, but I was alone in bringing much of the midwest territory into compliance with the contractual responsibilities that ETEC undertook.
I can only hope that this group will forgive my self-indulgence, but I have to point out that Earl has done nothing to refute the suggestions that I made. They still stand on their own. Earl's clarifications suggest that Ken Converse knows what he is doing, a subject that I never refuted. On the contrary, I have referred Ken to a number of people who have contacted me for service over the years for a variety of instruments. In fact, I will be packing up a Hitachi SEM in the next week that I have suggested Ken for the installation and continuing service of.
I stand by my comments on a fully-focusing WDS spectrometer being a different beast requiring a careful and informed hand for proper maintenance and operation. And I stand by my assertion that the ETEC WDS spectrometer in particular is an extremely touchy and sensitive instrument to properly calibrate. Once accomplished, however, the ETEC Autoscan mated with the ETEC WDS is a very effective and stable electron microprobe system, something I can attest to with my own personal instrument.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
-----Original Message----- } From: "earlw-at-pacbell.net"-at-sparc5.microscopy.com {"earlw-at-pacbell.net"-at-sparc5.microscopy.com} To: Allen R. Sampson {ars-at-sem.com} Cc: amenex-at-amenex.com {amenex-at-amenex.com} ; Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Hello Fellow Microscopist:
This is my first posting since joining the group. I have learned quite a bit just from reading your back and forth inquires.
I have a question of my own. A few years ago I got a Cambridge Stereoscan 120 SEM, but soon moved into another post and lost track of the vendor who sold the instrument to us. Now the SEM is in pretty bad shape and I would like to get in touch with the manufacturer, Cambridge Instruments - Electron Optics Group. So far I have tried without any success.
Does anyone of you know where or how to get in touch with these people? They don't appear in any listing or website I have look into.
Furthermore, does any of you by chance own an operational stereoscan instrument? Please reply.
-----Mensaje original----- De: BEN HIDALGO {bhidalgo-at-raudo.udo.edu.ve} Para: microscopy-at-msa.microscopy.com {microscopy-at-msa.microscopy.com} Fecha: domingo 18 de julio de 1999 1:54 Asunto: SEM- Need Help on Stereoscan
} Hello Fellow Microscopist: } } This is my first posting since joining the group. I have learned quite a bit } just from reading your back and forth inquires. } } I have a question of my own. A few years ago I got a Cambridge Stereoscan } 120 SEM, but soon moved into another post and lost track of the vendor who } sold the instrument to us. Now the SEM is in pretty bad shape and I would } like to get in touch with the manufacturer, Cambridge Instruments - Electron } Optics Group. So far I have tried without any success. } } Does anyone of you know where or how to get in touch with these people? They } don't appear in any listing or website I have look into. } } Furthermore, does any of you by chance own an operational stereoscan } instrument? Please reply. } } Thank you for your kind help. } } Ben Hidalgo, Ph.D. Materials Science Depart. Universidad de Oriente Venezuela E-mail: bhidalgo-at-ci.udo.edu.ve
I have an older (1991) stock of object slides from a German brand ("Assitent, ELKA slides, cleaned, with frosted stripe on both sides, no. 2406"). There's something strange with those: the've turned white/opaque and they all stick together.
Under the microscope, some kind of crystalisation process is visible, with feather-like crystals.
When I store the slides as I usualy do, in ethylalcohol 90%, with 3% hydrochloric acid (v/v), the crystals dissolve, but some kind of blue-metalic haze (in reflected light) stays on the slides and is impossible to remove. This haze is non-crystaline in appearence and is very noticiable in darkfield and phasecontrast.
Preparations I made with these slides (several 1000's!) seems all to be lost, as the mounting medium seems to have a habit to shrink excessively on those slides. Strangely enough this is only the case with slides made with Numount. Slides made with Entellan, Naphrax, Rhenohistol, Venetian turpentine and water based media (von Apathy's sirop, glycerinjelly) are OK.
In older literature (Langeron, M: "Precis de microscopie", 1934) I've found a possible explanation: Langeron speaks of an alteration of slides and coverslips, called "devitrification" (French). This alteration is/was not uncommon in a hot and humid climate but it happens/happened in all climates, only much faster under tropical conditions.
As possible solutions Langeron advises:
* sometimes the white opaque can be dissolved in acids (see above) * sometimes the white opaque can be dissolved in acid alcohol (see above) * the slides can be stored in paraffinoil or clove oil * the slide are protected when stored in air tight metal boxes * sometimes it's possible to clean the slides using FeCl3, 3% aq. (impossible, tried it). * always bye slides of premium quality (I tought "Assistent" was a top brand and it's widely used in Europe).
Does this sounds familiar to anyone? Possible solutions? Prevention?
Ben, Cambridge has been bought and sold several times over the last few years. They are currently called LEO. There general number is 800-356-1090 and web site is http://www.mwrn.com/leo/leo.htm. I hope this helps.
Brian Wajdyk
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Cambridge Instruments ha changed ownership several times in the last few years. They are currently LEO. LEO's general phone number is800-356-1090 and there web page is http://www.mwrn.com/leo/leo.htm. I hope this helps.
Brian Wajdyk
-- Brian Wajdyk, Senior Electron Microscopist, Center for Intergrated Systems Development, Motorola (FESEM / EDS / Scanning Auger Microscopy)
Cambridge Instruments ha changed ownership several times in the last few years. They are currently LEO. LEO's general phone number is800-356-1090 and their web page is http://www.mwrn.com/leo/leo.htm. I hope this helps.
Brian Wajdyk
-- Brian Wajdyk, Senior Electron Microscopist, Center for Intergrated Systems Development, Motorola (FESEM / EDS / Scanning Auger Microscopy)
I _desperately_ need information about the Princeton Gamma-tech 307 AD-converter NIM-module. I have recently acquired an old EDS system based on PGT electronics and I only have manuals for SiLi, spectroscopy amp and pileup/livetime-corrector. The AD converter apparently has some sort of fast sample-and-hold circuit (with calibration potentiometers at front) and a computer digital-io connection at back. I'd guess that the computer connection has 12 data pins (12bit converter) and some trigger/reading-good/whatever pins to control transfer (flow-control) of data.
The module was originally connected to PGT ancient inhouse-designed "computer" with ferrite-core memory.. I intend to connect the module to modern PC with windows software.
So, if someone has manual for this converter, or experience about it, please, let me know. I'm, of course, willing to pay reasonable cost for copies of manuals, airmail etc.
btw: I do have contacted both PGT directly and their Finnish representative. Their response was that they have _no_ support for these old modules and have dumped all their documentation. :(
I need to know if there is a japanese diamond knife and who is the manufactor and references about, if possible an E-mail for contact. I'll appreciate the help,
I must first apologize for this posting....I have for sale on ebay a Unitron binocular scope with camera. I started it at a buck, there is no reserve. I initially signed up to this list in an effort to figure out how to use/test the scope. I assume now that this is less a unit than most of you folks are working with, or want to deal with. Anyhow, Thanks for the fun...signing off now. http://cgi.ebay.com/aw-cgi/eBayISAPI.dll?ViewItem&item=133518177 -Matt B I can be contacted here or vovo-at-verinet.com _________________________________________________________ Do You Yahoo!? Get your free -at-yahoo.com address at http://mail.yahoo.com
Ben Hidalgo wrote: =====================================================
I have a question of my own. A few years ago I got a Cambridge Stereoscan 120 SEM, but soon moved into another post and lost track of the vendor who sold the instrument to us. Now the SEM is in pretty bad shape and I would like to get in touch with the manufacturer, Cambridge Instruments - Electron Optics Group. So far I have tried without any success.
Does anyone of you know where or how to get in touch with these people? They don't appear in any listing or website I have look into. ================================================ Their website URL is http://www.leo-em.co.uk/
Address information in the UK:
LEO Electron Microscopy Ltd Clifton Road Cambridge CB1 3QH England Telephone (44) 1223 414166 Fax (44) 1223 412776 E-mail info-at-leo-em.co.uk
Hope this helps!
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
{bold} Laboratory: {/bold} Electronic Materials and Engineering, Department = of Electronics, University of Barcelona.
{bold} Duration: {/bold} 12-18 months from January (or April) 1999.
{bold} Subject {/bold} : Structural and chemical characterization of UV-coati= ngs using TEM, XPS and XRD techniques, within the frame of a TMR project, in collaboration with LZH-Hannover, FHG-Jena, CEA- Grenoble, ENEA-Rome, FORTH-Crete and MSU- Moscou.
The aim of the project is to assess the effects of microstructure and composition inhomogeneities in the optical behaviour of the high reflectant or antireflectant coatings. Some of the materials involved are MgF2, LaF3, ... on Si, quartz and CaF2 substrates
{bold} Requirements: - {/bold} nationality from a {bold} EU member {/bold} (n= ot Spanish)
- Younger than 35 years
- PhD already finished or in the last period.
{bold} Merits: {/bold} - Experience in TEM and related techniques
{bold} Estimated Salary: {/bold} 1800 Euros/month
Applications including a CV, list of publications, and the names and addresses of at least two referees should be sent to:
I am new to the group but not a microscopist. My background is in photography and imaging, I currently work at the Materials Technology Laboratories in Ottawa, Canada, employed primarily to deal with digital images.
I have read the past correspondence and would like to offer my thoughts. As the topic seemed to be drifting toward quality I was surprised some standard control measures were not mentioned. Manipulation of a silver image is not limited to printing alone - development time, temperature, agitation, developer formula and concentration, even sharpening ( not limited to digital) can all factor into the quality of the resulting negative. Although I agree that digital images are no match for conventional film, they are close and getting closer. Using a service bureau to drum scan and output an image will reveal just how close. I've looked at an 8x10 camera original and it's digitally output twin through a 10x loupe and was surprised at the quality of the digital version. As a matter of fact, I would predict none but the very experienced could see a difference with the naked eye. The digital image was in the region of 6Gb. Hardly convenient, but the possibility exists to eventually come very close to film if not surpassing it in some areas. Generation degradation for one.
Archiving film is easy, relatively inexpensive and very long term with no degradation (if done properly). I haven't seen any studies that show a digital image would fare as well. What wasn't mentioned was discreet tonality, and I believe film wins hands down. Film's ability to record subtle shading (degree is format dependent) gives it a quality I have not seen in a digitally produced print. I don't believe digital will replace film, but it will win over a very large market share. In some areas it will dominate because of its cost effectiveness, i.e. press and catalogue photography. Keep in mind though, a skilled photographer with a digital back will more than likely produce a sharper image than a less skilled person using film. I can't speak for microscopists but there are many ways to produce a sharp image, it is not limited to a particular acquisition medium.
David Ashe NRCan / MTL Ottawa , Canada dashe-at-NRCan.gc.ca
There is a full time position available in The Jackson Laboratory's Biological Imaging Facility. Successful applicants must be proficient in state-of-the-art Light Microscopy (LM), TEM and SEM instrumentation use. Previous experience with standard EM and LM sample preparation and immunostaining techniques is critical. Basic darkroom and computer skills are also required; rudimentary programming skills are preferred. Applicants should have either a B.S. in a biological field with two years EM experience in a research setting or an M.S. in a biological field with one year of experience in EM. Courses at either the undergraduate or graduate level focusing on microscopy and immunohistochemical techniques would be an asset. This customer service driven position requires solid communication skills; applicants must be able to work independently in a multi-user facilty and interact with a variety of people. Friendliness and the ability to work as a team member are essential.
The Jackson Laboratory is one of the world's foremost centers for mammalian genetics research. Located in Bar Harbor, Maine, the lab is adjacent to Acadia National Park. Mountains, ocean, forests, lakes and trails are all within walking distance. If you love high tech challenges but you're looking for a more natural environment, this could be the opportunity you've been searching for.
Please contact: Human Resources The Jackson Laboratory 600 Main St. Bar Harbor, Maine 04609 e-mail: jobs-at-jax.org The Jackson Laboratory is an Equal Opportunity/Affirmative Action Employer. Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
South Bay Technology produces the PC2000 Plasma Cleaner which is based o= n technology that we license from Argonne National Laboratory. Technical information on Plasma Cleaning can be found on our web site at www.southbaytech.com. We have also recently introduced our PC2000 Plasm= a Cleaner which does not yet appear on our web site. I can send you a PDF version of that product brochure via email if that would be of interest. =
The PC2000 will be "officially" released at the upcoming MSA meeting in Portland. Please contact me off-line if you would like any additional details.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "paqui" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Dear All,
We are considering the possibility of up-date our equipment with a =
Plasma-Cleaner.
I would appreciate both commercial information and also (and =
specially) the opinion from the users of this kind of equipment.
Kind regards
Paqui
*******************************+ Francesca Peiro
EME, Electronic Materials and Engineering Dpt. Electronics =
University of Barcelona Marti i Franques 1 08028 Barcelona, Spain
Hi everyone I am spreading DNA using the droplet spontaneous adsorption technique. The DNA is spread fine, however, the Cytochrome C coating of the DNA is irregular- thick and thin, sometimes 2-3X the usual diameter, at other points not coating the DNA at all. The background is also irregular. My readings suggest the problem is with the ammonium acetate concentration, I have tried 0.25M, 0.30M and 0.45M, None of these produces a clear coating consistently. Any suggestions?
Sally Burns Center for Electron Optics B5 CIPS, MSU East Lansing, MI 48823 517 355-5004 burnssal-at-msu.edu
Their web site is (in the US): http://www.leo-usa.com/
good luck.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Garber, Charles A.[SMTP:CGARBER-at-2SPI.COM] } Sent: Monday, July 19, 1999 12:25:08 AM } To: MICROSCOPY BB } Subject: Where is LEO? } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Ben Hidalgo wrote: =====================================================
I have a question of my own. A few years ago I got a Cambridge Stereoscan 120 SEM, but soon moved into another post and lost track of the vendor who sold the instrument to us. Now the SEM is in pretty bad shape and I would like to get in touch with the manufacturer, Cambridge Instruments - Electron Optics Group. So far I have tried without any success.
Does anyone of you know where or how to get in touch with these people? They don't appear in any listing or website I have look into. ================================================ Their website URL is http://www.leo-em.co.uk/
Address information in the UK:
LEO Electron Microscopy Ltd Clifton Road Cambridge CB1 3QH England Telephone (44) 1223 414166 Fax (44) 1223 412776 E-mail info-at-leo-em.co.uk
Hope this helps!
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
I've been depressed because I can't afford a diamond knife and I've been told I would get much better results with my cardiac tissue using a diamond knife rather than a glass knife. I also just finished my thesis project which involved crustacean cysts. I put in so many hours at the microtome on that one!!
Anyway, I've been thinking that with all the recent advances in materials science, there's got to be a better way... Has anyone investigated whether a diamond-quality (or near diamond-quality) knife could be manufactured from a ceramic material? Anybody else have any thoughts on this topic?
-- Sonia Cawsey McGowan Copley Library, University of San Diego email: scawsey-at-acusd.edu home page: http://www.acusd.edu/~scawsey
I am seeking a JEOL 100CX TEM for parts (need not be operational) and will pick it up at your lab. Additionally, we are interested in the availability of a late model used (must be operational) 200kV analytical TEM for purchase. Please contact me off line at the address below.
Owen
============================= Owen P. Mills Electron Optics Facility Engineer Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
I have attempted to purchase a drawing tube from Zeiss to fit the old "standard" model scopes, but was dismayed to find out that it is no longer available. If anyone has one that is not being used and wishes to sell it, I would be very interested.
Richard A. Snyder, Ph.D. Center for Environmental Diagnostics and Bioremediation (CEDB) Biology Department University of West Florida 11,000 University Parkway Pensacola, FL 32514
http://www.uwf.edu/~rsnyder/
Work (850) 474-2806 FAX: -3130 _____________________________________________________________
} Anyway, I've been thinking that with all the recent } advances in materials science, there's got to } be a better way... } Has anyone investigated whether } a diamond-quality (or near } diamond-quality) knife } could be manufactured from } a ceramic material? } Anybody else have any thoughts on this topic?
Dear Sonia, Diamond is harder than any substance except boron nitride (I think), and, in addition to the hardness, the knife edge cannot flake off, so there is a stringent constraint on the crystal size and the binding of crystallites to one another if the knife is to work properly. Perhaps there is a way to produce a suitable ceramic knife edge, but I'd guess that this would involve epitaxial growth of the crystal to make the edge. If so, the knife would be much more expensive than diamond. Saphire (hardness 9 vs diamond 10 on the Moe (?) scale) is a material which is used for knives, and it may be suitable for your work. Good luck. Yours, Bill Tivol
I'm not sure I'd borrow a knife even if I could find a willing lender. I'd get too nervous about harming it. Fortunately, today I had some success with tissue that had been embedded using a slightly different protocol. But I'm thinking more about this from a broader perspective. As you mention, diamond knives have always been expensive. I don't think this alone, though, is enough to spur the development of a cheaper alternative. Market forces seem to be very quirky when it comes to anything to do with scientific products. I'm primarily a librarian, and journal price increases have been outpacing inflation since the 70's, but we still buy them because our patrons need them. I'd love to cancel the journals guilty of gouging, but often they are the most important (and the publishers know it). Another, more mundane example: I've found that a couple of 12 inch square pieces of 100 micron nylon mesh costs $75.00 if you buy it marketed for cell culture applications, but you can get a 1.5 x 5 foot bag filter for aquaculture applications made out of the same mesh
for $20.00. I'm not convinced that a cheaper alternative to diamond knives does not exist simply because it isn't possible. However, if alternative materials have been tried and have been found inadequate, then I guess I can get even more depressed.... (hmm ... where'd I put my stash of chocolates?)
"Malis, Tom" wrote:
} Sonia, I feel for your predicament, but I think that, in a sense you are } barking up the wrong tree, as the old saying goes. Diamond knives have been } around for nearly 40 years and have been expensive for all of that time, yet } no viable alternative has emerged. Sapphire knives are used on occasion } (there was just a Listserver inquiry as to where one could buy them), but } are still expensive, at about half the cost of diamond. So, there is just } about zero chance of anyone coming up with a ceramic alternative in the near } future. } } Why not focus your creative thinking in the other direction, which is to } find someone with a knife who is willing to let you borrow it? Microtomists }
} Tom Malis } Group Leader - Characterization } Materials Technology Laboratory } Natural Resources Canada (Govt. of Canada) } 568 Booth St., Ottawa, Canada } ph. 613-992-2310 } FAX 613-992-8735 } email: malis-at-nrcan.gc.ca {mailto:malis-at-nrcan.gc.ca} } } ---------- } From: Sonia Cawsey McGowan [SMTP:scawsey-at-acusd.edu] } Sent: July 19, 1999 12:42 PM } To: microscopy list } Subject: Alternatives to Diamond Knives } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } I've been depressed because I can't afford a diamond knife and } I've been told I would get much better results with my } cardiac tissue using a diamond knife rather than a glass knife. } I also just finished my thesis project which involved } crustacean cysts. I put in so many hours } at the microtome on that one!! } } Anyway, I've been thinking that with all the recent } advances in materials science, there's got to } be a better way... } Has anyone investigated whether } a diamond-quality (or near } diamond-quality) knife } could be manufactured from } a ceramic material? } Anybody else have any thoughts on this topic? } } -- } Sonia Cawsey McGowan } Copley Library, University of San Diego } email: scawsey-at-acusd.edu } home page: http://www.acusd.edu/~scawsey } }
-- Sonia Cawsey McGowan Copley Library, University of San Diego email: scawsey-at-acusd.edu home page: http://www.acusd.edu/~scawsey
I wonder if anyone can help me out. I'm processing some cell cultures through to LR White for immunocytochemistry. This is my current procedure;
Trypsinise (~ 4 min.) and scrape cells
Spin down to pellet at 1000 rpm in PBS
Fix in 2% paraformaldehyde, 0.05% glutaraldehyde for 2 hours
Buffer wash
Dehydrate 50 / 70 / 80 / 90 % Methanols at 4 degrees over ~3 hr
Put through 4 changes of LR White ( 1 hour each )
Embed at ~50 degrees C for 48 hours
Everything goes fine until the buffer wash after fixation . At this point my cell pellet does a near miraculous disappearing trick ! I am not flushing it into the waste. What then is happening ? The only thing I can think off is that the fixative is too weak. I am doing the whole thing again on Wednesday morning so any advice would be more than welcome. Yours Hopefully,
Barry Shaw
EM Technician, School of Biomedical Sciences, University of Nottingham Medical School
Hi everyone I am spreading DNA using the droplet spontaneous adsorption technique. The DNA is spread fine, however, the Cytochrome C coating of the DNA is irregular- thick and thin, sometimes 2-3X the usual diameter, at other points not coating the DNA at all. The background is also irregular. My readings suggest the problem is with the ammonium acetate concentration, I have tried 0.25M, 0.30M0. and 0.45M, None of these produces a clear coating consistently. Any suggestions? -- Sally Burns Center for Electron Optics B5 CIPS, MSU East Lansing, MI 48823 517 355-5004 burnssal-at-msu.edu
Listies: I need some info on sources for iridium foil. We used to get it from VCR Group, but my co-worker tells me he cannot contact them anymore. We use it in an ion sputter coater for SEM samples. Vendors' replies are quite welcome. We are in the USA, North Texas.
South Bay Technology, Inc. recently acquired VCR Group and is continuing = to manufacture their products - now under the South Bay Technology name. I will send you by separate email a copy of the Ion Beam Sputtering and Etching System price list for your review. The iridium target you are looking for is part no. 30.2090i. I will verify the price on that target=
and send it to you tomorrow.
If you plan to be at MSA, please stop by our booth (#518) so we can updat= e you on what is going on with South Bay Technology and VCR. As if gettin= g the SBT/VCR update isn't enough, we'll also be offering a free gift to th= e first 50 people who stop by to say hello!
In the mean time, if you have any questions concerning VCR Group product= s, please feel free to contact me directly.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Becky Holdford } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Listies: I need some info on sources for iridium foil. We used to get it from VCR Group, but my co-worker tells me he cannot contact them anymore. We use it in an ion sputter coater for SEM samples. Vendors' replies are quite welcome. We are in the USA, North Texas.
Can anyone suggest a Long Island, NY, SEM facility which offers machine time to experienced users in an academic environment or on a fee basis at low cost? EDS capability would be nice, but not essential.
We have been using cork/vial storage of SEM 1/8" pin mounts for a very long time. We punch a hole in the center of the small end of the cork to receive the pin and push stub on the small end of the cork into the vial. By using vials just slightly large in diameter than the stub, the assembly is quite compact. The vials are 1 dram shell vials, size 15 x 45 mm, manufactured by Kimble. The corks are VWR Laboratory corks size 4, regular length.
John Rensberger
On Thu, 15 Jul 1999, JIM ROMANOW wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } I have contacted five major U. S. SEM supply vendors and only two of them } sell 1/8" pin mount storage boxes that will accommodate high samples } (approx. 1/2" to 5/8" clearance). } The least expensive and highest box is out of stock for at least two weeks } and I need some ASAP. (The cost of these boxes range from 1.35 to 6.00 } dollars each!) Any suggestions would be appreciated. You can probably guess } who the vendors that I contacted are but I would be interested to find any } new players out there. Thank you. } } James S. Romanow } The University of Connecticut } Physiology and Neurobiology Department } Electron Microscopy Facility } U-131 } Storrs, CT 06269-2131 } bsgphy3-at-uconnvm.uconn.edu } 860 486-2914 voice } 860 486-1936 fax } } } }
Sonia - The discussion about the economics of diamond knives has missed an important point; most of the cost results from the labor involved, not the knife materiel itself. There's maybe $100 worth of diamond in a $1000 knife, plus a lot of highly skilled craftsman time. That's why the sapphire costs so much, relatively speaking; cheaper crystal, same labor. When you get beyond the thesis stage of your career, I hope that you can convince your employer that a diamond knife is actually a bargain compared to the productive hours (and irreplacable samples) lost with glass knives.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Francesca: Shop around and that would include looking at the EMITECH K1050x. You can read up about that online on our page K5 (enter from "Contents") Please note that PST cannot supply Emitech instruments outside Australasia and would not supply additional info outside our territory. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Monday, July 19, 1999 11:02 PM, paqui [SMTP:paqui-at-el.ub.es] wrote: } } } Dear All, } } We are considering the possibility of up-date our equipment with a } Plasma-Cleaner. } } I would appreciate both commercial information and also (and } specially) the opinion from the users of this kind of equipment. } } Kind regards } } Paqui } } } *******************************+ } Francesca Peiro } } EME, Electronic Materials and Engineering } Dpt. Electronics } University of Barcelona } Marti i Franques 1 } 08028 Barcelona, Spain } } Tel. (34-93) 402 11 39 } Fax. (34-93) 402 11 48 } e-mail: paqui-at-el.ub.es } **************************** }
Sonia - I fail to believe that muscle cannot be sectioned well with a glass knife. However, a diamond does make life easier. If I was a supervisor, I would be concerned to give a diamond knife to an inexperienced person - i.e. one who cannot section soft tissues with a glass knife. The cost of the diamond in a knife is significant, but smaller than the cost of manufacturing and testing, not to mention marketing. So it makes sense to use the best material available. Diamond is the hardest substance known and retains a superfine edge better than anything else. Tungsten carbide is next in hardness, but it has an intrinsic grain structure, making it only useful for sections above 1.5 um. Sapphire knives have been commercially available for over ten years. They have not made inroads. Probably because they have a much shorter life expectancy and only provide a modest saving. One could debate at which point is it worthwhile for a student to learn thin sectioning. The worst you can do is to become depressed. Reality is that you do not have a diamond knife available, that muscle can be sectioned with glass well and that you need to polish up on technique, unless somebody else is going to do the sectioning for you. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Tuesday, July 20, 1999 2:42 AM, Sonia Cawsey McGowan [SMTP:scawsey-at-acusd.edu] wrote: } } } I've been depressed because I can't afford a diamond knife and } I've been told I would get much better results with my } cardiac tissue using a diamond knife rather than a glass knife. } I also just finished my thesis project which involved } crustacean cysts. I put in so many hours } at the microtome on that one!! } } Anyway, I've been thinking that with all the recent } advances in materials science, there's got to } be a better way... } Has anyone investigated whether } a diamond-quality (or near } diamond-quality) knife } could be manufactured from } a ceramic material? } Anybody else have any thoughts on this topic? } } } -- } Sonia Cawsey McGowan } Copley Library, University of San Diego } email: scawsey-at-acusd.edu } home page: http://www.acusd.edu/~scawsey } }
We often look at paper samples but attach them directly to a brass stub using double- sided sellotape, coat them with gold-palladium and then image in SEM using secondary and/or backscattered electrons - any resin in the paper specimen is easily distinguishable from paper fibres. We don't embed our paper in any sort of resin
Best regards
Belinda
Belinda White Senior EM Technician Centre for Electron Microscopy University of Natal Pietermaritzburg South Africa
} I wonder if anyone can help me out. I'm processing some cell } cultures through to LR White for immunocytochemistry. This is my } curren t procedure; } } Trypsinise (~ 4 min.) and scrape cells } } Spin down to pellet at 1000 rpm in PBS } } Fix in 2% paraformaldehyde, 0.05% glutaraldehyde for 2 hours } } Buffer wash } } Dehydrate 50 / 70 / 80 / 90 % Methanols at 4 degrees over ~3 hr } } Put through 4 changes of LR White ( 1 hour each ) } } Embed at ~50 degrees C for 48 hours } } Everything goes fine until the buffer wash after fixation . At this } point my cell pellet does a near miraculous disappearing trick ! } I am not flushing it into the waste. What then is happening ? The } only thing I can think off is that the fixative is too weak. I am } doing the whole thing again on Wednesday morning so any advice would } be more than welcome. Yours Hopefully, } } Barry Shaw } } EM Technician, School of Biomedical Sciences, } University of Nottingham Medical School }
Hi Barry: This is what I would do:
Fix cells with double strength fixative added to the medium at a ratio of 1:1 (8 % formaldehyd and 0,4 % ga is often used) Leave for 1-2 hours. Carefully scrape the cells with a piece of Teflon cut to form a blade (a soft wooden stick will also do the job) and pellet the cells by centrifugation in the presence of a small amount of protein (FCS or BSA 1-2%). The protein will prevent the cells from sticking to the wall of the tube, something that might happen. You can also embed the cell pellet in 10 % gelatin in water and threat the cells as tissue during further processing:remove the supernatant, resuspend the pellet in 100 ul 10 % gelatin, centrifuge and leave on ice for 1 hour, cut the tip off the tube and remove the gelatinembedded pellet that can be cut into small blocks containing cells.
For further details about embedding etc I suggest you look into
Are there any Transmission Electron Microscopists (especially Zeiss TEM users) out there who have converted their TEM to enable digital photography?
We are interested in bolt-on systems such as the Megaview II, and would be very interested to hear from electron microscopists who have already got these systems installed........
Of particular interest: how well does the resolution and blackness of the blacks of digital micrographs compare to those printed from traditional b/w film?
Amanda Wilson St George's Hiospital Medical School awilson-at-sghms.ac.uk
At 09:42 AM 7/19/1999 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Barry, Regarding your problem with disappearing cell culture pellets: Try spinning your sample down before each change. Then resuspend the cells each time to make sure they're exposed evenly to each solution and not packed in the bottom of the tube. Good luck, Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
} Listies: I need some info on sources for iridium foil. We used to } get it from VCR Group, but my co-worker tells me he cannot contact } them anymore. We use it in an ion sputter coater for SEM samples. } Vendors' replies are quite welcome. We are in the USA, North Texas.
Dear Becky, Alfa Aesar sells Ir foil of several different thicknesses. You can reach them at (800) 343-0660, or www.alfa.com. The only connection I have to the company is that I get their catalog. Yours, Bill Tivol
Job Posting: Scientist in the Microscopy Group of the Analytical and Materials Characterization Laboratories of Polaroid Corp., R&D Chemical Research Division, Waltham, Ma.
* B.S. or M.S. in Materials Science * Strong communication skills, written and oral * Works well in a team environment * Strong light microscopy skills * Hands on experience in a variety of other microscopy and materials characterization techniques desirable e.g. scanning electron microscopy, transmission electron microscopy, microtomy, x-ray photo-electron spectroscopy, x-ray diffraction, energy dispersive x-ray analysis, probe microscopy, * Photonic materials experience is a plus * Creative, problem solver * Additional knowledge in analytical techniques valuable
If interested, Pls. contact Lynne Garone, Manager of Analytical and Materials Characterization Labs 1265 Main St. W4 -1D Waltham, Ma. 02451-1714 phone #: (781) 386-1446, Fax # (781) 386-0378, e. mail: GaroneL-at-Polaroid.com
It is inexpensive to use glass for knives, but the glass degrades repidly. Has anyone used the resin additves which are supposed to extend glass knife edge life?
Bill
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
JEOL has asked for permission to post the following message. Since it is an open invitation to all microscopist's attending the upcoming meeting in Portland, and has been made in the spirit of good will and camaraderie, I consider it appropriate to pass along to the listserver community.
Nestor Your Friendly Neighborhood SysOp
================================================================= To All M&M '99 Attendees:
We at JEOL are celebrating our 50th anniversary this year and are holding a reception at the Microscopy & Microanalysis 1999 Conference to commemorate this event. This reception will be held immediately following the exhibit hours on Tuesday, August 3, 1999 in the Ballroom at the Oregon Convention Center. The same location as the sessions.
We hope that you will set aside this time to come and visit with us and enjoy some refreshments. For those of you attending the MAS Presidential Symposium we hope that you will be able to join us at 6:00 PM.
Free passes to this reception will be available to everyone at the conference during exhibit hours on Monday and Tuesday. Please feel free to bring family and friends.
Again, it's in the Ballroom at the Oregon Convention Center on Tuesday from 5:00 until 8:00 PM.
If you have any questions please contact Steve Hamilton at 978/536-2270 or hamilton-at-jeol.com. ==================================================================
Steve Hamilton Tel: 978/536-2270 JEOL USA, Inc. Fax: 978/536-2401 11 Dearborn Road Email: hamilton-at-jeol.com Peabody, MA 01960 WWW: http://www.jeol.com
================================================================== Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-5075, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov ================================================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ==================================================================
The box said "This program requires Win 95/98/NT or better..." so I bought a G3 Mac
Dear Becky, VCR has been recently acquired by SouthBay Technology and Vince Carlino is at SouthBay, so you could try them. Another good source for all metals is Goodfellow: www.goodfellow.com. At 06:49 PM 7/19/99 -0500, you wrote: } } Listies: I need some info on sources for iridium foil. We used to } get it from VCR Group, but my co-worker tells me he cannot contact } them anymore. We use it in an ion sputter coater for SEM samples. } Vendors' replies are quite welcome. We are in the USA, North Texas. } } -- } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Becky Holdford (r-holdford-at-ti.com)
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
POSTDOCTORAL POSITION AVAILABLE TO STUDY MICROTUBULE ORGANIZING CENTERS (MTOCs; CENTROSOMES) IN CANCER CELLS AND IN A TRANSGENIC ADENOCARCINOMA MOUSE PROSTATE MODEL
A postdoctoral position is available immediately to study centrosome-cytoskeletal interactions in cancer cells and tissue. The successful candidate will join an established research lab and an established group of investigators at the Cancer Research Center to study centrosome organization and abnormalities in cancer cells using novel microscopy methods and low voltage field emission scanning electron microscopy (LVFESEM).
Our lab was among the first to identify centrosomes with immunofluorescence microscopy and to characterize centrosome structure with high resolution scanning and transmission electron microscopy. Monoclonal antibodies against centrosomes have been raised to identify tissue specific centrosome proteins. We are currently interested in abnormal centrosome formation that is associated with multipolar spindle organization and genomic instability.
Qualifications for this position will include basic experience with tissue culture cells, and knowledge in light and electron microscopy. Salary will be commensurate with experience. Please submit a curriculum vitae, the names and addresses of three references, and a brief statement of research interests to:
Heide Schatten, Ph.D. Associate Professor Department of Veterinary Pathobiology University of Missouri-Columbia 1600 E. Rollins Street, Columbia, MO 65211 TEL: (573) 882-2396 FAX: (573) 884-5414 e-mail: SchattenH-at-missouri.edu
SELECTED RELATED PUBLICATIONS:
Schatten, H., Schatten, G., Mazia, D., Balczon, R., and Simerly, C. Behavior of centrosomes during fertilization and cell division in mouse oocytes and in sea urchin eggs. Proc. Natl.Acad. Sci. USA 83: 105-109 (1986).
Schatten, H., Walter, M., Mazia, D., Biessmann, H., Paweletz, N., Coffe, G., and Schatten, G. Centrosome Detection in Sea Urchin Eggs with a Monoclonal Antibody against Drosophila Intermediate Filament Proteins: Characterization of the Division Cycle of Centrosomes. Proc. Natl. Acad. Sci. USA 84:8488-8492 (1987).
Schatten, H. Dithiothreitol Prevents Membrane Fusion but not Centrosome or Microtubule Organization During the First Cell Cycle in Sea Urchins. Cell Motil. Cytoskel. 27, 59-68 (1994).
Thompson-Coffe, C., Coffe, G., Schatten, H., Mazia, D., and Schatten, G. Cold-Treated Centrosome: Isolation of Centrosomes from Mitotic Sea Urchin Eggs, Production of Anticentrosomal Antibody, and Novel Ultrastructural Imaging. Cell Motil. Cytoskel. 33, 197-207, 1996.
Petzelt, C., Werner, D., and Schatten, H. In Vivo-Labeling of the Centrosome of Human Primary Endothelial Cells by Transfection with a Green Fluorescent Protein-Centrosomin A Construct. Mol. Biol. of the Cell, (1997).
Schatten, H. and Chakrabarti, A.,. Centrosome Structure and Function is Altered by Chloral Hydrate and Diazepam During the First Reproductive Cell Cycles in Sea Urchin Eggs. Eur. J. Cell Biol., 75, 9-20.
Chakrabarti, A., Schatten, H., Mitchell, K.D., Crosser, M., and Taylor, M. Chloral hydrate alters the organization of the ciliary basal apparatus and cell organelles in sea urchin embryos. Cell Tissue Res. 293, 453-462 (1998).
Schatten, H., Hedrick, J., and Chakrabarti, A. The cytoskeleton of Drosophila-derived Schneider line-1 and Kc23 cells undergoes significant changes during long-term culture. Cell Tissue Res. 294, 525-535 (1998).
Schatten, H., Ripple, M., Balczon, R., and Taylor, M. Centrosome abnormalities in cancer cells and tissue. ICEM14, 243-244 (1998).
Schatten, H., Ripple, M., Balczon, R., Taylor, M., and Crosser, M. Centrosome proliferation in the human androgen-responsive LNCaP and the androgen-independent DU 145 prostate cancer cell lines. Proceedings MSA 4(2), 1066-1067 (1998).
Schatten, H., Chakrabarti, A., and Hedrick, J. Centrosome and microtubule instability in aging Drosophila cells. J. Cellular Biochemistry 74:229-241 (1999).
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sonia,
You say that you cannot obtain useful sections of cardiac muscle using = glass knives and are looking for an alternative to diamond knives. =
Here are two suggestions that may help you out:
First, have you really tried to cut good sections on the glass knives? = Sometimes this defeats people because they embed their samples in a resin = formulation that is just too hard to cut using glass knives. Try = embedding in a soft Spurr resin formulation and cutting that. I am sure = it will work.
The second suggestion is that you look at the special resharpening offers = available from some of the diamond knife suppliers. Basically, if you can = get your hands on any old diamond knife, you can trade it in for a new one = at a little over half the price.
If you are really stuck on getting good sections come up to Los Angeles = for a day and I will let you use one of my diamond knives.
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
by imo23.mx.aol.com (IMOv20.25) id fAVEa10245 (14386) for {microscopy-at-msa.microscopy.com} ; Tue, 20 Jul 1999 14:40:51 -0400 (EDT) Message-ID: {65b53d99.24c61cb9-at-aol.com}
I agree with Paul,glass knives can be used to produce consistently very good sections. However,a softer Spurr or Epon formulation must be used. For 8 years,we used this technique in a commercial veterinary pathology lab on all types of animal tissue with very good results. The only time we had to use the diamond knife was when we received hard blocks from customers. The trade off is that the images were not quite as crisp as those produced by using a diamond knife on a hard block, but they were still very useable for all purposes. Hope this helps Norm Woodside
JEOL has asked for permission to post the following message. Since it is an open invitation to all microscopist's attending the upcoming meeting in Portland, and has been made in the spirit of good will and camaraderie, I consider it appropriate to pass along to the listserver community.
Nestor Your Friendly Neighborhood SysOp
================================================================= To All M&M '99 Attendees:
We at JEOL are celebrating our 50th anniversary this year and are holding a reception at the Microscopy & Microanalysis 1999 Conference to commemorate this event. This reception will be held immediately following the exhibit hours on Tuesday, August 3, 1999 in the Ballroom at the Oregon Convention Center. The same location as the sessions.
We hope that you will set aside this time to come and visit with us and enjoy some refreshments. For those of you attending the MAS Presidential Symposium we hope that you will be able to join us at 6:00 PM.
Free passes to this reception will be available to everyone at the conference during exhibit hours on Monday and Tuesday. Please feel free to bring family and friends.
Again, it's in the Ballroom at the Oregon Convention Center on Tuesday from 5:00 until 8:00 PM.
If you have any questions please contact Steve Hamilton at 978/536-2270 or hamilton-at-jeol.com. ==================================================================
Steve Hamilton Tel: 978/536-2270 JEOL USA, Inc. Fax: 978/536-2401 11 Dearborn Road Email: hamilton-at-jeol.com Peabody, MA 01960 WWW: http://www.jeol.com
================================================================== Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-5075, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov ================================================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ==================================================================
The box said "This program requires Win 95/98/NT or better..." so I bought a G3 Mac
I once saw a small lightbox that has a footprint a little larger than a notebook. The transparent window is angled at about 30 degrees. If you are using Neg-a-file sheets for your negatives, this lightbox sits in your notebook inserted between pages and a sheet of negatives of interest can be viewed.
Does anyone know who makes this thing and where to buy it?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Barry: Cell pellets can be treated like bits of tissue if they are near 1mm thick. If you have less material it will disappear by re-suspending. Then it would need to be spun during every step - a method to be avoided.
Fix pellet for 5 to 10 minutes and then use a needle to gently score a one mm square pattern through the pellet. Complete the fixation time. 120 minutes with 2.5% of fixative is rather long for cells - half an hour (in the cold) should be plenty. After that, gentle handling will keep the cells clumped and they can be treated like a bit of tissue. Keep the cells in the original tubes during processing and gently add and withdraw the various liquids.
Cells fixed in suspension will not remain clumped after pelleting. By not scoring the pellet early during fixation, fixative penetration and forming of small clumps are impaired. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Tuesday, July 20, 1999 9:53 AM, BARRY SHAW [SMTP:Barry.Shaw-at-nottingham.ac.uk] wrote: } } I wonder if anyone can help me out. I'm processing some cell } cultures through to LR White for immunocytochemistry. This is my } current procedure; } } Trypsinise (~ 4 min.) and scrape cells } } Spin down to pellet at 1000 rpm in PBS } } Fix in 2% paraformaldehyde, 0.05% glutaraldehyde for 2 hours } } Buffer wash } } Dehydrate 50 / 70 / 80 / 90 % Methanols at 4 degrees over ~3 hr } } Put through 4 changes of LR White ( 1 hour each ) } } Embed at ~50 degrees C for 48 hours } } Everything goes fine until the buffer wash after fixation . At this } point my cell pellet does a near miraculous disappearing trick ! } I am not flushing it into the waste. What then is happening ? The } only thing I can think off is that the fixative is too weak. I am } doing the whole thing again on Wednesday morning so any advice would } be more than welcome. Yours Hopefully, } } Barry Shaw } } EM Technician, School of Biomedical Sciences, } University of Nottingham Medical School } }
Scott D. Walck wrote: ======================================================= I once saw a small lightbox that has a footprint a little larger than a notebook. The transparent window is angled at about 30 degrees. If you are using Neg-a-file sheets for your negatives, this lightbox sits in your notebook inserted between pages and a sheet of negatives of interest can be viewed.
Does anyone know who makes this thing and where to buy it? ======================================================= There is probably more than one manufacturer of this sort of thing, but we have offered for some time the Slim Edge® Light Pad. It has an 8x10" viewing area and has overall dimensions of 12.25 x 9.25". It is only ˝" thick. More information can be found on URL http://www.2spi.com/catalog/photo/lightpad.html
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
Thank you very much for all the helpful comments, suggestions and links that you sent me. I appreciate it very much.
Claudia Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT12 2EE ++44(0)181 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
Could you please refer on a provider of low-melting point solders, about 40-60C of melting point. If it possible to make a proper alloy from pure Ga and In by myself, I'd be happy to know the composition of it. The alloy must be inert for HF acid, or at least not to contaminate the surface of the sample after deepening into HF for tens of seconds. The solder is necessary for InP-based samples mounting onto magnetic holders for works with conductive probe AFM. I hope to reduce sample drift together with good conductivity of the joint. If you know a better solution for the drift-free conductive sample mounting, avoiding oxidation of the sample surface, I would be glad to here it from you!
Best regards. Dmitri.
__________________________________________ Dmitri V. Sokolov, Doctor Course student Research Center for Interface Quantum Electronics, Hokkaido University, North 13, West 8, Kitaku, Sapporo 060, Hokkaido, Japan Phone 81-11-706-7174 Fax 81-11-716-6004 http://www.geocities.com/SiliconValley/Campus/1314 AOL Instant Messenger FalconDot ICQ 9418072 __________________________________________
You may have to consider fixing in the flask if all else fails or at least some form of encapsulation such as in agar if it doesn't. interfere with your immuno work. Most references I have seen also emphasize that fixative and buffer solutions need to be decanted with great care down the sides of the centrifuge tube to avoid dispersing the pellet.
My experience has mainly been with histological fixation (2.5% glutaraldehyded) of culture cells which always worked well if the cells were fixed in the flask but on the one occasion a research assistant scraped then fixed we ended up with small pellets of disrupted cells. The only problem may be lifting the fixed cells off the flask surface - for this you may have to experiment to find the optimum time after fixation is started.
Good luck
Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
---------- } From: BARRY SHAW To: Microscopy
I wonder if anyone can help me out. I'm processing some cell cultures through to LR White for immunocytochemistry. This is my current procedure;
Trypsinise (~ 4 min.) and scrape cells Spin down to pellet at 1000 rpm in PBS Fix in 2% paraformaldehyde, 0.05% glutaraldehyde for 2 hours Buffer wash Dehydrate 50 / 70 / 80 / 90 % Methanols at 4 degrees over ~3 hr Put through 4 changes of LR White ( 1 hour each ) Embed at ~50 degrees C for 48 hours
Everything goes fine until the buffer wash after fixation . At this point my cell pellet does a near miraculous disappearing trick ! I am not flushing it into the waste. What then is happening ? The only thing I can think off is that the fixative is too weak. I am doing the whole thing again on Wednesday morning so any advice would be more than welcome. Yours Hopefully,
Barry Shaw
EM Technician, School of Biomedical Sciences, University of Nottingham Medical School
Hi everyone =20 =20 I am using the IMQuant software (Oxford Instruments) to characterise=20 carbide particles in steel. However, the samples analysed are 2-D=20 sections and diameters or areas of carbides are understated compared=20 with the 3-D reality. Is there a mathematical way of compensating for=20 this 2-D limitation within this software? =20 Also, what's the best resolution I can achieve, is it related to the=20 SEM resolution? =20 Many thanks =20 F.
} Dear All, } } We are considering the possibility of up-date our equipment with a } Plasma-Cleaner. } } I would appreciate both commercial information and also (and } specially) the opinion from the users of this kind of equipment. } } Kind regards } } Paqui } } } *******************************+ } Francesca Peiro } } EME, Electronic Materials and Engineering } Dpt. Electronics } University of Barcelona } Marti i Franques 1 } 08028 Barcelona, Spain } } Tel. (34-93) 402 11 39 } Fax. (34-93) 402 11 48 } e-mail: paqui-at-el.ub.es } **************************** } Dear Paqui,
We are a UK based manufacturer of plasma barrel reactors that may suit your purpose. A local distributor based in Barcelona is: Leica Microsistemas C/Nicaragua, 46 08029 Barcelona Tel: +34 93 494 95 30 Fax: +34 93 494 95 32 E-mail:ana.alarcon-at-leica-microsystems.com Contact: Ana Alarcon
Best regards Mike Wombwell Polaron range Business Manager V G Microtech The Birches Industrial Estate Imberhorne Lane East Grinstead West Sussex RH19 1UB UK Direct line: +44 (0)1342 310296 Switchboard: +44 (0)1342 327211 Fax: +44 (0)1342 315074 http://www.polaron-range.com E&OE
Spreading DNA is a very tedious procedure. I am not familiar with the spontaneous adsorption technique, but I know the spreading technique has always worked for me and I'm including it here. But first, let me ask you several questions. What buffer are you using and at what pH? This is very important. A more alkaline solution has always worked for me (see below). Second, what is the purpose of ammonium acetate? I have never used this and achieved excellent results. Third, what is your hypophase solution? This is very important since this serves as a matrix. Look at my procedure. It is a very modified one and I cannot locate the original source right now.
Spreading solution: Working Solution:
1 M Tris buffer - pH 8.5 + 0.1M EDTA 10ul 99% formamide - highly purified molecular bio. grade 50ul
DNA - (50ug/ml stock solution) 2ul ddH2O - 33ul cytochrome C 5ul always add last
Add in order given. Make sure that any glass used, including slides must be soaked in HCl/Chromic acid solution for 2 days prior to use. Also, as you know,everything must be DNAase free.
Hypophase - floating solution.
ddH2O 79ml 1M Tris (above) + 0.1M EDTA pH 8.5 1ml formamide 20ml The procedure is the slide spreading technique. The DNA is collected onto carbon-coated grids 400m. The next step was to stain in 0.05M uranyl acetate in 95% ethanol and 0.1M HCl.
Please give me a call if you have any questions and I would be more than happy to explain further the technique. Blessings................................................................... Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Hoechst Marion Roussel, Inc. 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-hmrag.com
I have been working for some time to produce sections of an extremely fragile medium and have at last succeeded. However, this has meant that Osmium Tetroxide has had to be used in the fixation process along with freeze substitution. I am embedding in Lowicryl. Does anyone have any information on immunolabelling of Osmicated sections?
Thanks ********************************************************************** Caroline Routledge Department of Optometry and Vision Science University of Wales, Cardiff Redwood Building King Edward VII Avenue Cardiff South Glamorgan
Bill: Never noticed any difference with the additives. I even tried the tungsten coating technique (evaporate W onto the glass knife) with variable results. I have found that the original Luft (medium) formulation to be the most consistent and to provide the best results with glass knives.
Roger Moretz Dept of Toxicology
Opinions expressed are my own and do not reflect those of my empoyer.
} -----Original Message----- } From: William McManus [SMTP:billemac-at-biology.usu.edu] } Sent: Tuesday, July 20, 1999 9:21 AM } To: 'microscopy-at-Sparc5.Microscopy.Com' } Subject: RE: Alternatives to Diamond Knives } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } } It is inexpensive to use glass for knives, but the glass degrades repidly. } Has anyone used the resin additves which are supposed to extend glass } knife } edge life? } } Bill } } William McManus } Supervisor } Electron Microscopy Facility } Department of Biology } Utah State University } Logan UT 84322-5305 } } billEMac-at-cc.usu.edu } 435-797-1920
Fischione Instruments(www.fischione.com) makes an excellent plasma cleaner. We have a dry-pumped system that has proven itself very valuable for cleaning TEM samples and our various holders, as well as bulk SEM samples, aperatures, etc. Both prevention of contamination and removal of previous contamination have been excellent.
There was a very good paper in Microscopy and Microanalysis (March/April '99) by Isabell, et. al. dealing with the plasma cleaner and it's applications/results for electron microscopy. Worth reading, especially if you're considering a purchase.
Regards,
Mike Coy ************************************************************************ Mike Coy m-coy-at-nwu.edu SEM Facility Manager (847)491-3439 Electron Probe Instrumentation Center (EPIC) Northwestern Unversity ************************************************************************
In a message dated 7/21/99 8:33:10 AM, frank.sarrazit-at-AVESTASHEFFIELD.COM-at-sparc5.microscopy.com writes:
} I am using the IMQuant software (Oxford Instruments) to characterise } carbide particles in steel. However, the samples analysed are 2-D } sections and diameters or areas of carbides are understated compared } with the 3-D reality. Is there a mathematical way of compensating for } this 2-D limitation within this software?
I don't know that particular software package but probably the best solution is to measure a size distribution of the two-dimensional feature sections, say in 10 to 15 bins, and write that data out to Excel. Performing the matrix multiplication to get the diameters of the three-dimensional features that correspond to the measured section areas is trivial there. Matrices corresponding to spheres and ellipsoids can be found in Weibel's 1980 Stereological Methods book (Academic Press) and in my 1986 Practical Stereology book (Plenum) among others. For other shapes (cubes, tetrahedra, etc.) there is a harder-to-find book by J. Wasen and R. Warren (A Catalogue of Stereological Characteristics of Selected Solid Bodies) published in 1990 by Chalmers Univ. in Sweden (my copy does not show any isbn number). When using this method, be aware that it is mathematically "unstable" - i.e., the statistical precision of your final result is much worse than the counting precision of the number of features in each classification bin - and that the results depend very much on having the correct shape assumption about your particles.
It is clear that the Microscpopy Community needs a simple forum where Commerical Organizations can post News Items of interest. We have discouraged this practice on the Microscopy Listserver, however, the need still exists nevertheless.
As such, in the infinite amount of spare time available to me, I have created an experimental WWW site which will allow commerical organizations to post messages, announcements etc.. This WWW site is independent of the Microscopy Listserver, but still maintained on the MSA servers. The URL is:
and is accessible via the MSA Home Page as well as a variety of links at other WWW sites.
A few points to bear in mind.
Firstly, any commerical organization who fills out the Electronic Submission Form will be permitted can post to the WWW Electronic News Server. However, in order to maintain a degree of content oversight, no posting will be uploaded to a viewable page without having been reviewed first. This is to attempt to insure that proper decorum is maintained. I (for the moment) will review the text of submissions as they are posted and if it in my judgement they are not bogus and are appropriate then I will uplink them to the Active News/Announcement WWW pages. If I find something that appears out of line, then I will of course contact a vendor.
Second, no editorial action will be taken on the postings. They will appear as submitted. I will not have time to be an editior, so for those of you who make use of this system, be forewarned. Read your postings carefully before submitting them!
Third, after some finite period of time the postings will be culled to remove old or outdated information.
The Microscopy Listserver will still operate as it always has, but I hope this will provide our (important) commerical members a mechanism to provide timely information to our scientific community. It will, I hope, remove the frustration I know alot of commerical vendors have with the rather strict rules about no advertising on the Microscopy Listserver.
As this is an experiment, so please bear with any growing pains. If it becomes to much of a problem then we will try some other mechanism but at least it is an attempt. My one request is that you please use it in the spirit it is intended.
Cheers....
Nestor Your Friendly Neighborhood SysOp.
================================================================== Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-5075, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov ================================================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ==================================================================
The box said "This program requires Win 95/98/NT or better..." so I bought a G3 Mac
Our Core users have been generating zigabytes of data on our = deconvolution and confocal microscopes. We have found, unfortunately, = the CD writing interfaces we have on these systems are too slow and = complicated (or error prone) for most users. As a result we have been = forced to spend several hours a week writing CD's for people. It takes = about a 30 minutes/CD: 15 min data prep and 15min actual writing. My = question, finally, is what are other labs charging to write CDs for=20 their users. =20 Thanks,
Hank Adams Integrated Microscopy Core Baylor College of Medicine Soggy Houston
} Members, } } Our Core users have been generating zigabytes of data on our deconvolution } and confocal microscopes. We have found, unfortunately, the CD writing } interfaces we have on these systems are too slow and complicated (or error } prone) for most users. As a result we have been forced to spend several } hours a week writing CD's for people. It takes about a 30 minutes/CD: 15 } min data prep and 15min actual writing. My question, finally, is what are } other labs charging to write CDs for } their users.
Hank,
We charge for a half-hour of technician time but it does not really take that long. We use a 4X Yamaha writer and Adaptec software on an NT workstation. It is a very simple process to drag the files into the data window of the CD Creator software and click the red button. We charge for time at the worstation at $5/hr plus the tech time. I wonder if the problems you are encountering are from using older software (some of the early stuff was dreadful) or older hardware?
Good luck.
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 530 754 7536 fax raharris-at-ucdavis.edu
Members, I appreciate all the replies I received so far on CD writing. I = guess I should have added that about 15 gigs of data is generated/wk and = we are writing from SGI unix machines. We actually have few problems = with WinNT data transfer from users-either Zip (I know not the best), CD = writing or FTP from our users.=20
Thanks,
Hank Adams Integrated Microscopy Core Baylor College of Medicine Sweltering Houston
This is an old thread which came up about 2 years ago, however, since it has reappeared, it is appropiate to point out that Mike Coy neglected to mention that in addition to Fischione, a number of other companies namely- South Bay Technology (http://www.southbaytech.com) and Structure Probe Inc (http://www.2spi.com) also sell and market plasma cleaners for AEM applications in the USA. There is also a variation which is being developed by XEI (http://www.msa.microscopy.com/SM/XEI/XEIHomePage.html) for in-line SEM applications.
I also use this methodology routinely. I happen to use the SBT model and can echo Mike Coy's exact comments which apply equally well to the SBT unit, it has " proven itself very valuable for cleaning TEM samples and our various holders, as well as bulk SEM samples, aperatures, etc. Both prevention of contamination and removal of previous contamination have been excellent. "
The SBT unit at ANL is used on instruments including Philips, JEOL, Hitachi, AEI and VG covering the gamit of operating modes .. AEM, SEM, CTEM, STEM, IVEM,HREM, and HVEM.
I can testify to the technologies effectiveness in literally all operating conditions. An important point to remember is that this methodology works most effectivley when the contamination source is specimen borne (a key point that most people forget to mention). If you have a "dirty" instrument to start with , then cleaning the specimen while ignoring the column will not be very effective. Of course, modern instruments have relatively low inherent contamination rates, so the newer the instrument the more effective the technology.
Nestor Your Friendly Neighborhood SysOp
Disclaimer: ======== Although I am a satisfied user, I must disclose that my employer Argonne National Laboratory holds the original patent on this technology and recieves royality for units sold based upon that patent. Each of the above manufacturer's are licensee's of the ANL Patent.
I am asked to look for morphology and composition by SEM/EDS of some strongly ferro-magnetic powders, about 20=B5m in size. We would like to avoi= d embedding and cross-sectionning.
=46rom experience, they tend to sink in epoxy during curing when we glue the= m on a substrate. Did somebody try to use ferro-magnetic stubs (particles should stick to the closest ferromagnetic surface, but astigmatism=8A)?
Any suggestions how to mount the powder on the stub owing to the magnetic forces which will attract the particles to the pole-pieces?
Best regards
Philippe
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
Philippe Buffat wrote: ========================================= I am asked to look for morphology and composition by SEM/EDS of some strongly ferro-magnetic powders, about 20µm in size. We would like to avoid embedding and cross-sectionning.
} From experience, they tend to sink in epoxy during curing when we glue them on a substrate. Did somebody try to use ferro-magnetic stubs (particles should stick to the closest ferromagnetic surface, but astigmatismŠ)?
Any suggestions how to mount the powder on the stub owing to the magnetic forces which will attract the particles to the pole-pieces? ================================================= The SPI Supplies Tacky Dot™ Slides might work just fine for this application. If you select the 15 µm dot slide, only one particle will stick (and quite tenaceously) to each "dot". Wait 24 hours after application for the bond to develop to its maximum strength. It would be possible for the local field strength to be sufficiently high that the strong adhesive bonds could be overcome and some or all of the particles could get pulled off of their respective "dots"on the slide.
More information about Tacky Dot Slides can be found on URL http://www.2spi.com/new/tacky.html
Dsiclaimer: SPI Supplies manufactures Tacky Dot Slides under license from E . I. du Pont de Nemours and Co., Inc.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
Alternatives to Diamond Knives, response to Sonia Cawsey's message
In the mid '70s, Ian Roberts described a method for coating glass knives to prolong their cutting life and, although they will never match a diamond knife for sustained sectioning, may well offer a solution to sectioning difficulties experienced with conventionally prepared glass knives. As I have always been fortunate enough to have a good diamond knife for sectioning, I have not tried coated glass knives myself. However, I have had it on good authority that the coating works and have kept this reference safely tucked away in case I ever needed it: Roberts, I M (1975), J Microsc 103 113-119: Tungsten coating of glass knives.
Sonia Cawsey McGowan wrote:
I've been depressed because I can't afford a diamond knife and } I've been told I would get much better results with my cardiac tissue using a diamond knife rather than a glass knife. I also just finished my thesis project which involved crustacean cysts. I put in so many hours at the microtome on that one!! Continues ...
SEM Stub Storage Boxes, response to James S. Romanow's message
I have not used commercially-available SEM stub storage boxes for many years now, ever since a colleague showed me an alternative:
We use storage boxes manufactured from rigid, transparent plastic (we call them sandwich boxes), which are available in different depths. Their bases can be lined with a thin sheet of expanded polystyrene or I prefer to use stiff card (Ilford paper boxes are ideal) cut to size and with the edges turned down to support the sheet at the desired height. Suitably spaced holes are then punched in the polystyrene/card for holding the stubs. A refinement is to cover the top of the polystyrene/card with some squared/graph paper, not only does this help with the layout of holes but also allows you to write labels etc.
These boxes are cheap, hold lots of samples, are very adaptable for different samples, are stable and, if the type with a push-in lid is used, do not suffer from broken hinges. Obviously, for tall samples you need to use deeper boxes.
James S. Romanow wrote:
I have contacted five major U. S. SEM supply vendors and only two of them } sell 1/8" pin mount storage boxes that will accommodate high samples (approx. 1/2" to 5/8" clearance). The least expensive and highest box is out of stock for at least two weeks } and I need some ASAP. (The cost of these boxes range from 1.35 to 6.00 dollars each!) Any suggestions would be appreciated. You can probably guess who the vendors that I contacted are but I would be interested to find any new players out there. Thank you.
I hope that these suggestions may be of help.
Regards,
Jill Webb Reading Scientific Services Ltd, Reading, England
I use the ImQuant package on our system, and I do not believe that there is a way to do the transformation within the program. I use the Display/Excel function within the Analysis windows to transfer the data to Excel and then do my processing to sort the features into size bins of my own design.
Frankly, I don't much care for trying to massage the data back into its "true" 3-D distribution. Like John Russ said, these can be rather unstable exercises. Sure, there is some underestimation of true size when analyzing the 2-D section, but how critical is the true size? Can you not get good enough information from a comparison of 2-D results for different samples? I guess I am more interested in a good engineering solution now than in the "right" scientific answer.
Warren
At 12:09 PM 7/21/1999 +0000, you wrote: } Hi everyone } } } I am using the IMQuant software (Oxford Instruments) to characterise } carbide particles in steel. However, the samples analysed are 2-D } sections and diameters or areas of carbides are understated compared } with the 3-D reality. Is there a mathematical way of compensating for } this 2-D limitation within this software? } } Also, what's the best resolution I can achieve, is it related to the } SEM resolution? } } Many thanks } } F.
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
Over the last year, it has become increasingly difficult to turn the specimen injector rod in the airlock of our Philips 201 TEM ( no service contract) when inserting or removing specimens. Would anyone know what the cause might be, and if I could cure it during routine maintenance/cleaning of the microscope? The rod itself does not appear to be bent and the specimen holder fits snugly onto its tip. I have cleaned the rod and speciment holders, but I don't know if something needs lubrication and I've been afraid of introducing any chemicals into the column. Any suggestions would be appreciated. Regards,
Andrew Ochalski, Microscopy Technician, Dept. of Biology, University of Ottawa Room 108, Gendron Bldg. 130 Louis Pasteur, Ottawa, Ontario CANADA K1N 6N5 613-562-5800 x6343 FAX: 613-562 5486
I didn't "neglect" anything. I simply gave an opinion on a piece of equipment that I use daily and am very happy with at the request of a fellow microscopist looking for information. I have never used SBT's or Structure Probe's units, and thus didn't offer an opinion of them.
SBT had previously responded to this thread, as well as another manufacturer, without mentioning all other units. Were those "neglectful" as well?
Regards,
Mike Coy
************************************************************************ Mike Coy m-coy-at-nwu.edu SEM Facility Manager (847)491-3439 Electron Probe Instrumentation Center (EPIC) Northwestern Unversity ************************************************************************
Alternatives to Diamond Knives, response to Sonia Cawsey's message
In the for what it's worth dept.
Back in the early to mid 80's I worked for Reichert-Jung (it was another life), and an artical crossed my desk wherein they (no I don't remember who'they' are) used vitrious carbon sheets which were broken like glass knives. The quality of which was compared to diamond knives. I believe that the artical was published in BioTechniques. This is really reaching into the dim recesses of my memory and I do not remember if anything else was done with it.
best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
Don't go around saying the world owes you a living; the world owes you nothing; it was here first. Mark Twain [Samuel Langhornne Clemens] (1835-1910)
urgent: Is there anybody from near Frankfurt/Germany who is joining the MSA MAS meeting in Portland / Oregon in the beginning of August? I would like to send a poster there and need someone to take it with them. Thank you for your help.
Alexander Du Chesne, M.S., Ph.D. Max-Planck-Institut für Polymerforschung PF 3148, 55021 Mainz Tel. 0049 6131 379 195 Fax 0049 6131 379 100 E-mail: duchesne-at-mpip-mainz.mpg.de
Dear Philippe, My experience with imaging iron powder is that on an SEM/EDX and with such small particle size, there should not be any problem if you use a small sample and stick it down well. I recommend the carbon double-sided sticky tabs, which have a good stick. Use a small sprinkle of powder and press it well into the sticky tab. Using magnetic stubs will cause bad astigmatism in the SEM. I found out the hard way not to put magnetic powders into the TEM, but I have lovely pictures of them stacking up in spiral towers in the field of the objective lens. Then they stuck to the polepiece and it was a service call to get any imaging back. At 03:33 PM 7/21/99 +0200, you wrote:
} Hello all, } } I am asked to look for morphology and composition by SEM/EDS of some } strongly ferro-magnetic powders, about 20=B5m in size. We would like to= avoid } embedding and cross-sectionning. } } From experience, they tend to sink in epoxy during curing when we glue them } on a substrate. Did somebody try to use ferro-magnetic stubs (particles } should stick to the closest ferromagnetic surface, but astigmatism=8A)? } } Any suggestions how to mount the powder on the stub owing to the magnetic } forces which will attract the particles to the pole-pieces? } } Best regards } } Philippe }
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
We have had a lot more success with glass knives using the 55 degree angle setting. It involves a major of the resetting the microtome, but the percentage of usable knives went up considerably than when we used the "standard" 45 degree angle. The hardest part was breaking the taboo of violating the Tribal Knowledge. Also, when making the final break, do it very slowly. In fact, if you can turn the breaking knob in little moves and allow the break to occur "on its own" seems the best. A friend suggested this over ten years ago and it made a big difference.
Mike Baxter mykkb-at-juno.com
___________________________________________________________________ Get the Internet just the way you want it. Free software, free e-mail, and free Internet access for a month! Try Juno Web: http://dl.www.juno.com/dynoget/tagj.
Andrew, Your problem is either a maladjustment on the airlock, or the grease inside the specimen airlock tube is 'thickening' up and not lubricating as well as it should. I was with Philips for 14 years, now an independent, and have worked on/serviced dozens of EM201s. If you'd like you could either e-mail or call me and I could suggest several things to do, or clean to get the airlock functioning properly. Regards, Ron Veil V.E.I.L.(Veil Electron Instrument Lab.) Customer Services veilcs-at-earthlink.net tel.(650) 952-3099 FAX (650) 869-4978
On Thu, 22 Jul 1999 09:44:24 GMT+5 "Andrew Ochalski" {aochalsk-at-science.uottawa.ca} writes: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America
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although i know it is a longshot, i was wondering if anyone happens to have a GATAN TV CONTROL UNIT MODEL #622-0600 that you may want to sell. it interfaces to a fiber optic camera for TEM, although it may be used in other optical applications; i don't know. although our unit still works, the power supply is on its way out and gatan is charging an outrageous price to repair it. thanks for your time.
paul
-------------------
Paul E. Anderson Catalytic and Nanostructured Materials Laboratory Department of Chemistry 102 Hurting Hall Northeastern University Boston, MA 02115 (617) 373 5909 FAX: (617) 373 8795 paanders-at-lynx.neu.edu
} } } "Andrew Ochalski" {aochalsk-at-science.uottawa.ca} 07/22/99 09:44am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Andrew, After thoroughly cleaning the injector rod, did you apply any vacuum grease to the o-ring? Lubricate the o-ring with some Philips-recommended grease. It's worked for me the past 20 years! Bob Santoianni Emory University Hospital Atlanta, Georgia -----------------------------------------------------------------------.
Fellow Microscopists,
Over the last year, it has become increasingly difficult to turn the specimen injector rod in the airlock of our Philips 201 TEM ( no service contract) when inserting or removing specimens. Would anyone know what the cause might be, and if I could cure it during routine maintenance/cleaning of the microscope? The rod itself does not appear to be bent and the specimen holder fits snugly onto its tip. I have cleaned the rod and speciment holders, but I don't know if something needs lubrication and I've been afraid of introducing any chemicals into the column. Any suggestions would be appreciated. Regards,
Andrew Ochalski, Microscopy Technician, Dept. of Biology, University of Ottawa Room 108, Gendron Bldg. 130 Louis Pasteur, Ottawa, Ontario CANADA K1N 6N5 613-562-5800 x6343 FAX: 613-562 5486
Stain Technol 1983 May;58(3):143-51 Vitreous carbon: a new material for making microtome knives. Disharoon DR, Wickham MG, Worthen DM, Lofftus FH
In the abstract (haven't gotten the article yet) they state that it can be broken like glass - so no big labor cost. I know it isn't the material itself that is the big cost. But if another material could be used that doesn't require so much labor...
Has anyone tried vitreous carbon? What are the reasons for it not catching on?
================================== Alternatives to Diamond Knives, response to Sonia Cawsey's message
In the for what it's worth dept.
Back in the early to mid 80's I worked for Reichert-Jung (it was another
life), and an artical crossed my desk wherein they (no I don't remember who'they' are) used vitrious carbon sheets which were broken like glass knives. The quality of which was compared to diamond knives. I believe that the artical was published in BioTechniques. This is really
reaching into the dim recesses of my memory and I do not remember if anything else was done with it.
best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
-- Sonia Cawsey McGowan Copley Library, University of San Diego email: scawsey-at-acusd.edu home page: http://www.acusd.edu/~scawsey
We are scanning in SEM negs using a high resolution scanner (Polaroid SS45) and have a problem with negs that show moderate to high contrast.
When images are captured and opened in Photoshop 4.0, we can adjust the gamma to get either excellent details in the shadows OR highlights but unfortunately the other one then suffers. Is there a way of adjusting for highlights and THEN shadows and somehow combining the best of both adjustments into one image?
Or am I asking for something beyond the capabilities of the program?
Thanks very much.
John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I am using a Zeiss microscope with a HBO 100 mercury arc bulb. It is used approximately 12 hours a day. Over time (weeks to months), a film develops upon the condenser lense that has brownish rainbow like pattern. This results in longer integration times for our CCD camera and general degradation of image quality. Has anyone noticed this on their systems, and is there a method to prevent it? Does anyone know the source of this film?
Thanks,
Jason Bush
_______________________________________________________________ Get Free Email and Do More On The Web. Visit http://www.msn.com
} } } } "rschoonh-at-sph.unc.edu"-at-sparc5.microscopy.com wrote: } } } } } Alternatives to Diamond Knives, response to Sonia Cawsey's message } } } } } } In the for what it's worth dept. } } } } } } Back in the early to mid 80's I worked for Reichert-Jung (it was another } } } life), and an artical crossed my desk wherein they (no I don't remember } } } who'they' are) used vitrious carbon sheets which were broken like glass } } } knives. The quality of which was compared to diamond knives. I } } } believe that the artical was published in BioTechniques. This is really } } } reaching into the dim recesses of my memory and I do not remember if } } } anything else was done with it. } } } } } } } I think this paper was in Stain Technology about 1982. } } } } 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 } } **********************************************
I scan with a gamma of between 1.6 and 2. This is primarily based on info that I got from a book entitled "Real World Photoshop 4". Something about the linear response of the detectors in the scanners to light - I forget the explanation. works well on a flatbed too. I adjust the exposure control on the SS45 from about -10% to about +15% depending on the negative. I also always scan in 12 bit resolution and adjust the levels in Photoshop just before I change the image to an 8-bit. I get a lot of control on the final image this way. You can set up an action that automates the levels, and switching to an 8-bit. I also always check the levels just before I switch to 8-bit just as a check.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: bozzola-at-siu.edu To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Digital Imaging Gurus,
We are scanning in SEM negs using a high resolution scanner (Polaroid SS45) and have a problem with negs that show moderate to high contrast.
When images are captured and opened in Photoshop 4.0, we can adjust the gamma to get either excellent details in the shadows OR highlights but unfortunately the other one then suffers. Is there a way of adjusting for highlights and THEN shadows and somehow combining the best of both adjustments into one image?
Or am I asking for something beyond the capabilities of the program?
Thanks very much.
John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Photoshop does the basic tasks pretty well. I suggest you to try the Curves and Levels tools (located in Image/Adjust menu) to equalize gray shades. The curves tool is my favorite. Using it, you can manipulate the shape of the gamma curve of an image. The best way is to start by pressing the auto button in the Curves dialog box, which will give you a basic tonal adjustment. Next, you can manipulate the shape of the gamma curve with the mouse to achieve the result that you want. You can also use the eyedropper icons in the same dialog box to select the wanted pure black and pure white that in the picture.
A more sophisticated image analysis package (analySIS) can be found at: http://www.soft-imaging.de/ . It can make miracles with some pictures, but I guess that it is somewhat expensive and not so friendly user as Photoshop. Anyway, if you need more, take a look at it.
Photoshop does the basic tasks pretty well. I suggest you to try the Curv= es and Levels tools (located in Image/Adjust menu) to equalize gray shades. = The curves tool is my favorite. Using it, you can manipulate the shape of the gamma curve of an image. The best way is to start by pressing the auto button in the Curves dialog box, which will give you a basic tonal adjustment. Next, you can manipulate the shape of the gamma curve with th= e mouse to achieve the result that you want. You can also use the eyedroppe= r icons in the same dialog box to select the wanted pure black and pure whi= te that in the picture.
A more sophisticated image analysis package (analySIS) can be found at: http://www.soft-imaging.de/ . It can make miracles with some pictures, bu= t I guess that it is somewhat expensive and not so friendly user as Photoshop. Anyway, if you need more, take a look at it.
S=E9rvio T=FAlio Pires Amarante
serviopa-at-usp.br
Museu de Zoologia da Universidade de S=E3o Paulo Caixa Postal 42694-970 04299-970 S=E3o Paulo BRASIL
Thanks to everyone who offered advice on how to fix my problems with LR White LM immunolabeling and regular TEM - especially Hildy Crowley.
I haven't yet improved the LM immunolabeling, but the TEM is much better.
I switched from dehydrating to 96% EtOH to 100% EtOH.
I infiltrated in the refrigerator rather than room temperature (3 changes, 40 min each) I filled the vials to the top of the container (no air space).
I used a "heat sink" during polymerization (50 deg. for around 20 hrs.) Since I don't have a machine shop (a heat sink can be made by drilling holes in an aluminum block) I made one out of stacked 1/4" and 15/16" zinc nuts. I glued 6 stacks together in a hexagon with nail polish. I filled in the bottom nuts with tin foil, put a coil of copper wire in the center and wrapped the outside with tin foil. I wrapped the 00 size gelatin capsules in a single layer of aluminum foil for a snug fit.
================== Another matter: I sent a post re: vitreous carbon this afternoon and it still hasn't shown up on the list. Is it possible it got rejected by the SPAM filter? (I have not received a message to that effect, though).
Does anyone have information on the State University of New York SEM classes? I tried to find information on the current status of these classes (I attended one many years ago) for a new employee we are training, but have not been successful. Any information would be appreciated.
Maybe not... Problem is probably in the barrel that the rod turns in.
Sadly, one must split the microscope above the specimen stage, lift the stage out, and then undo two screws that hold the barrel in place. Remove, clean and regrease the barrel. BUT, if there is excessive wear from the stage alignment rod (the rod gizee that moves the stage that is driven by an ecentric on the barrel), it's time to buy parts from your friendly Philips provider.
Good luck!
Ron
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
"Robert Santoianni" {Robert_Santoianni-at-emory.org} on 07/22/99 04:29:59 PM
To: aochalsk-at-science.uottawa.ca cc: microscopy-at-sparc5.microscopy.com
} } } "Andrew Ochalski" {aochalsk-at-science.uottawa.ca} 07/22/99 09:44am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Andrew, After thoroughly cleaning the injector rod, did you apply any vacuum grease to the o-ring? Lubricate the o-ring with some Philips-recommended grease. It's worked for me the past 20 years! Bob Santoianni Emory University Hospital Atlanta, Georgia -----------------------------------------------------------------------.
Fellow Microscopists,
Over the last year, it has become increasingly difficult to turn the specimen injector rod in the airlock of our Philips 201 TEM ( no service contract) when inserting or removing specimens. Would anyone know what the cause might be, and if I could cure it during routine maintenance/cleaning of the microscope? The rod itself does not appear to be bent and the specimen holder fits snugly onto its tip. I have cleaned the rod and speciment holders, but I don't know if something needs lubrication and I've been afraid of introducing any chemicals into the column. Any suggestions would be appreciated. Regards,
Andrew Ochalski, Microscopy Technician, Dept. of Biology, University of Ottawa Room 108, Gendron Bldg. 130 Louis Pasteur, Ottawa, Ontario CANADA K1N 6N5 613-562-5800 x6343 FAX: 613-562 5486
Had a major camera jam in a Hitachi H-7000. Now we are in need of about 25 film cassettes. The 7000 "dispenser" box either held 22 or 50 cassettes. The 22 size are much thicker and therefore much more robust. The thinner cassettes where 50 will fit in the box are the ones we are in need of. Does anyone know of a source (other than Hitachi) or are willing to part with some of these? Thanks.
Joel McClintock U of Kentucky (606)257-1242 jmcclin-at-pop.uky.edu
The brown pattern you see on your collector lens is made by excess of Heat. HBO and XBO burners have this disadvantage to generate a lot of heat. This problem can be also seen by other types of microscopes. However, the lamphous from Carl Zeiss is particulary sensitive, because it is very small.
I have a company dedicated to service and repair of microscopes. The browning of collector lens is very current. Even, sometimes, the lens is cracked.
I have a solution for this. I built an automatic cooling fan. If you need more information, please send me a direct email at: emeylan-at-csi.com
best regards,
Emile Meylan SERCO Technical Services, Inc.
----- Original Message ----- } From: Jason Bush {jbec26-at-hotmail.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, July 22, 1999 5:10 PM
VACANCY - NEUROANATOMICAL SCIENTIST
The Neuroimaging Department at ReNeuron is responsible for the exploration of the spatio-temporal fate and the integration into host neural networks of neural cells transplanted into a number of models of neurodegenerative diseases.
We are immediately seeking a BSc/MSc graduate in Neuroscience or a related discipline to fill a vacancy in the Neuroimaging department. You will be expected to participate in experimental work using a number of histological and neuroanatomical techniques as well as computerized analysis of images from tissue sections. Previous experience is not essential as training will be provided for the successful applicant but an interest in data handling, graphics and computing is essential.
Please send your full career details either by email to Sara-Patel-at-reneuron.com or by post to Sara Patel, ReNeuron Ltd, Institute of Psychiatry, De Crespigny Park, Denmark Hill, London, UK, SE5 8AF.
I am constantly being impressed by the quality and quantity of generosity offered via this listserver. Thank you to all who sent the helpful suggestions regarding the gamma adjustment. I will pursue each one of the suggestions and respond individually. I look forward to meeting some of you at M&M next week in Portland. Again, thanks.
John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Hyrax: Custom research and Designs 8500 Mount Vernon Road Auburn california 93603
Phone: 916* 885-3341 *I think the area code has changed to 530.
Kim
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Kimberly A. Riddle Florida State University tel: 850.644.6519 Biological Science Imaging Resource 119 Bio Unit I, 4370 fax: 850.644.0481 Tallahassee, FL 32306 riddle-at-bio.fsu.edu ~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Dear colleagues Maybe I will unsubscribe soon ... I had many questions, but seldom an answer. That=B4s because I=B4m one o= f the "juniors" . Thanks especially to Hildy Crowley and Charles Garber, they are very active in this forum and had good hints regarding embedding for immuno-electron-microscopy (e.g. LR-White). Now, I=B4m doing quite good work in this special discipline and its possible that you will notice it one day. Recently, themes have changed in this listserver. Costs of digital imaging archives, salaries, used devices to sell or search and special advertisements, job offers seem to be the overwhelming topics nowadays. I=B4ve noticed that "greenhorn questions" remained unanswered in the firs= t half of the year ... As I am resident in Europe, offers of used devices and so on don=B4t interest me that much as the most of you all (in the U.S.). Thus, many messages from the listserver are not so interesting for me especially. Dear colleagues, why don=B4t you discuss more about techniques and methods? I am young and want to gain experience. Sure, digital archives of EM-photographs are now high-sophisticated but my intsitution won=B4t afford it in the near future. Eyeryone of us has the fear to talk about unpublished results or datas, but I feel that electron-microscopic techniques are so complex that they need a wide cooperation.
Thank you for your support. I will return to the listserver, that=B4s sur= e
Michael Reiner, cand. med Department of Anatomy I University of Colgne Germany Joseph-Stelzmann-Str. 9 50931 Koeln/Cologne Germany e-mail: a2811111-at-smail.uni-koeln.de Tel: +49-221-478-5519 Fax: +49-221-478-6411
Hello Greg! I feel you haven=B4t understand my intentions. I found acces to this listserver by way of the archives you named. I got useful hints from the archives and decided to participate in the listserver. I am not critizising the support that the MSA Listserver provides, sure it is the most advanced forum in special microscopy ! But I=B4m referring to the shift of themes and topics discussed in the listserver and the obviously ignorance of basic themes ( embedding, proceedings). Costs of new imaging and archiving possibilities, offers of postdoctoral=B4s and used devices seem to be overwhelming in the recen= t past. Everyone of us has knowledge in microscopy. We should share it, unpublished or not. Greetings. a young microscopist. Michael Reiner
Let me offer a couple quick thoughts and suggestions in response to your thoughts. I looked through my trash can and only found one other message from you, and that one was an answer to someone else's question. Besides, I am interested in materials more than biological samples, so most of your posts were quickly discarded.=20
I typically try to offer comments in areas where I have some knowledge, as do many users. But because of the traffic, I will often quickly delete the posts that are unrelated to my field(s). You may want to make your subject fields clear and concise.=20
Some posts will go unanswered because they the questions seem at such a rudimentary level. Someone asking about the right conditions to do such and so might be better off looking in a text first or taking some more training and then bringing a more specific question to the list. I don't think most listers care to do lots of training on line. But even so, I have seen a number of what I thought to be fairly general and rudimentary questions get answered on this list.=20
Similary, other posts will go unaswered because they are so general or vague. If someone were to ask for replies from users involved in image analysis, I doubt that they would get very many replies just because of the unspecific question. Some might offer a reply saying "I do image analysis. So what?" They really don't have much to go on to give a more helpful answer. However, if I were to ask for replies from those doing image analysis on SEM images of concrete, I think the response would be better. Subscribers would have less tendency to think that someone else would respond to this still pretty vague request. If I share some more details of my work at the outset and a particular question, I think listers are more likely to reply.=20
I offer these as general suggestions to all subscribers. Like I said, I don't remember the particulars of your posts. And sometimes, listers post questions that just don't have answers out there (like when I asked for those who had M-O drives that could read 650 MB cartridges from an HP drive). The silence itself is an answer. So I hope you will stick around and that these ramblings have been helpful.=20
WS
At 10:01 PM 7/23/1999 +0200, you wrote: } Dear colleagues } Maybe I will unsubscribe soon ... } I had many questions, but seldom an answer. That=B4s because I=B4m one of } the "juniors" . } Thanks especially to Hildy Crowley and Charles Garber, they are very } active in this forum and had good hints regarding embedding for } immuno-electron-microscopy (e.g. LR-White). } Now, I=B4m doing quite good work in this special discipline and its } possible that you will notice it one day. } Recently, themes have changed in this listserver. Costs of digital } imaging archives, salaries, used devices to sell or search and special } advertisements, job offers seem to be the overwhelming topics nowadays. } I=B4ve noticed that "greenhorn questions" remained unanswered in the first } half of the year ... } As I am resident in Europe, offers of used devices and so on don=B4t } interest me that much as the most of you all (in the U.S.). Thus, many } messages from the listserver are not so interesting for me especially. } Dear colleagues, why don=B4t you discuss more about techniques and } methods? } I am young and want to gain experience. Sure, digital archives of } EM-photographs are now high-sophisticated but my intsitution won=B4t } afford it in the near future. } Eyeryone of us has the fear to talk about unpublished results or datas, } but I feel that electron-microscopic techniques are so complex that they } need a wide cooperation.
We have the following items that are now surplus to our requirements in our lab.
1. A Gatan model 622 image intensified TV camera designed to mount on a JEOL 4000EX (i.e. we have the mounting flange for that instrument and the camera is shielded for use on that instrument. This camera is about 11 years old.
2. A Gatan model 673 wide angle TV camera that fits in the 35mm camera port of a JEOL 2000FX. This camera is over 8 years old.
3. Two Tracor TN5500 XEDS systems. a. One system has a 30Meg hard disk drive, two 5.25 Syquest removable hard disks (both failed) and two floppy disks one 5.25" and one 8". There are actually two 5.25" disks and two 8" disks in a separate subsystem, but the hard ware only supports two floppies at one time and so we have one of each set up. A standard Tracor keyboard with keypad and monitor is supplied. The system does not have a printer. We modified it so it would run without a printer and if we need print out we have a couple of switch boxes that directs the print out to a Mac. We also have the HP plot software and this is directed to a program on the Mac that can then send the plot to a laser printer or can save it for pasting into word processing documents. The system has the imaging package that will allow the computer to control the microscope (it is setup for a JEOL 2000FX) and record STEM and SEM images and XEDS mapsThe software includes SMTF and SQMTF. The system has an almost new refurbished light element detector (detects down to C). System also has a license for RT-11, the DEC operating system and it can run an FTP server for removal of spectra and images to a remote computer.
b. The second system is floppy based and also has imaging which is setup for an SEM whose manufacturer evades my memory, but if anyone is interested I will obviously find out for you. This system has a Be window XEDS detector with it.
4. Gatan 666 PEELS with mount for JEOL 2000FX. This system can be driven with the TN5500 (see 3a above) or with a Mac, the Mac is to be preferred and will be included Mac includes the data acquisition card from Gatan. This is one of the earliest 666 systems ever installed and is therefore about 10 years old.
5. Gatan single-tilt liquid-nitrogen cryo-transfer stage for Philips CM series microscope, I believe this is called a model 626.
6. JEOL straining stage for JEOL 2000FX
7. Two Gatan double-tilt Be cup analytical stages for the JEOL 2000FX, I think these are 646 models.
8. One thousand degree hot stage for JEOL 2000FX Gatan single tilt Model 62= 8
9. Liquid nitrogen cold stage for JEOL 2000 FX Gatan double tilt old model 613 upgraded to double tilt. Sample airlock pumps dewar jacket.
=46or further info just give me a call or drop me email.
Please note new FAX number.
John Mansfield PhD CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
I have a customer who wants to scrap a running JEOL JSM-U3 SEM. I suggested donation to the Smithsonian. It is free for anyone who wants it for parts or whatever.
} } Digital Imaging Gurus, } } We are scanning in SEM negs using a high resolution scanner (Polaroid SS45) } and have a problem with negs that show moderate to high contrast. } } When images are captured and opened in Photoshop 4.0, we can adjust the } gamma to get either excellent details in the shadows OR highlights but } unfortunately the other one then suffers. Is there a way of adjusting for } highlights and THEN shadows and somehow combining the best of both } adjustments into one image? } } Or am I asking for something beyond the capabilities of the program?
The task of adjusting different parts of an image independenly and then recombining them are variously described as "local" or "adaptive" algorithms. Since you want to modify contrast, most folk do it via direct manipulation of the histogram, though some do it in image space itself -- such as a proprietary "hysteresis" method or some greyscale mathematical morphology methods.
Unless your problem is extreme, try simple things first. Look for "local" or "adaptive" histogram methods: local histogram strech, adaptive histogram equalization, etc. Code for most of these are available on the web. I don't know what's available for Photoshop as a plugin, but I do know that some are available free for the freeware Photoshop work-alike called GIMP.
This is a first announcement to inform you of an upcoming conference....The First International Conference on Advanced Materials Processing, Rotorua, New Zealand, 19-23 November 2000.
Scope of the Conference:
In the last few decades of the 20th century, research in advanced materials processing has made a tremendous contribution to the development of novel, high performance and low cost materials. Without any doubt, in the new millennium, advanced materials processing will continue to be one of the most active research areas in materials science. To celebrate the historical achievement of scientists and engineers in this crucially important area, we are organizing the {bold} 1st International Conference on Advanced Materials Processing (ICAMP) in the very first year of the new millennium.
Sponsored by the Institute of Materials, UK, this conference will provide a forum for international researchers to exchange their recent findings and views in the area of advanced materials processing. Internationally renowned experts in this area will be invited to give keynote lectures to the conference.
During the conference, a workshop will also be organized to discuss opportunities for international co-operation in research on advanced materials processing.
Key dates:
Preliminary Title/Expression of Interest: Sept. 1 1999
Second Circular: October 1 1999
Abstract due by March 1 2000
The proceedings will be published in a hard-bound special edition book, and all papers will be fully refereed to ensure that they are of the highest standard.
The conference will be held in Rotorua, New Zealand. Rotorua is in the central North Island, and holds many attractions, including the world famous thermally active reserves.
Areas of Interest include:
Casting
Electronic materials
Extrusion
Filament Winding
Fuel cells
Hot compaction
HIP
Joining
Mechanical Alloying
Moulding
Powder Metallurgy
pre-Preg Production
Pultrusion
Rapid Solidification
Recycling
Repair
Sintering Processes
Solid State Chemistry
Tape Casting
Thermoset Matrix Composites
Thin Film Deposition
Traditional Ceramic Processes
Other
International Advisory committee:
Professor Aldinger (MPI, Germany)
Professor Boivin (USTL, France)
Dr. Bossel (Switzerland)
Dr Dicks (BG, UK)
Prof. Dunlop (Univ. Queensland, Australia)
Prof. Hanson (Riso Denmark)
Prof Hu (Institute for Metals Research, China)
Mr Jessup (CSIRO, Australia)
Prof Kilner (I.C., UK)
Prof Koch (North Carolina State Univ., USA)
Dr Lewin (BNFL, UK)
Prof McCormick (Univ. W. Australia)
Prof Muddle (Monash Univ., Australia)
Assoc. Prof. Petric (McMaster Univ., Canada)
Dr Pugh (Concordia Univ., Canada)
Dr Suzuki (NRIM, Japan)
For further information (and to receive the first circular), please contact:
Professor Nigel Sammes, Chair ICAMP 2000, n.sammes-at-waikato.ac.nz
or
Dr Deliang Zhang, Secretary, ICAMP 2000, d.zhang-at-waikato.ac.nz
Professor Nigel Sammes Department of Materials and Process Engineering The University of Waikato Private Bag 3105 Hamilton New Zealand
----- Original Message ----- } } Unless your problem is extreme, try simple things first. Look } for "local" or "adaptive" histogram methods: local histogram } strech, adaptive histogram equalization, etc. Code for most } of these are available on the web. I don't know what's available } for Photoshop as a plugin, but I do know that some are available } free for the freeware Photoshop work-alike called GIMP.
Just in case most of you are famailiar with "gimp" being a linux program, it has been ported to win32. I have just today realized this, and am thusly unfamiliar with its hardware compatibility and stability, but it is available for free download at: http://www.gimp.org/~tml/gimp/win32/ ... I am unfamiliar with the plugins mentioned above being available for this version.
A really good place to start is the "Tips & Tricks" archive I maintain. It is an archive of many of what "I" consider useful biologic discussions posted to the miocroscopy listserver. It is a bit of a chore and thus I am about a year behind, but there is still a fair ammount of info in there. If there was a discussion posted you recall but don't find, let me know as I have roughly 7megs ( about 200 discussions ) worth of material sitting in my mail and I can forward relevant hits to you.
The address is:
http://www.biotech.ufl.edu/~emcl
Follow the tips link
Nestor maintains a full archive of this list indexed by month at the address below.
http://www.msa.microscopy.com/
He is probably more current than I.
Good luck
At 10:01 PM 7/23/1999 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
{} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} Scott Whittaker ph: 352-392-1184 EM Technician fax: 352-846-0251 University of Florida email: sdw-at-biotech.ufl.edu ICBR EM Core Lab web:www.biotech.ufl.edu/~emcl/
OK - I can't stand it - I'm going to hop in on this one.
These techniques discussions don't happen in a vacuum - someone has to start them by asking a question or an opinion.
I don't think that anyone has the time (or the ego) to sit down at the computer and compose a "What I know about technique X" e-mail out of the clear blue. This would be the only other way to have the kind of information that you want make it out to the server.
Don't unsubscribe simply because the recent threads haven't been to your liking - you might miss some great stuff! The same thing happens on any listserver - there will be periods when nothing comes out that applies to your own work. Tough. Start a discussion of your own or just wait it out :) Besides - a lot of the "this isn't my area" stuff can suddenly BE in your area, then you wish you'd been paying attention! (personal experiene on that one - who knew we'd accidentally buy the wrong color CDs?!)
Allen R. Sampson 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 } -----------------------------------------------------------------------. } } Glad to hear it, Earl. I had never heard from either Hank or Ken that they } had been properly trained in WDS, and still have not. To the contrary, } after I first came on board, received orientation in Danbury for a week and } received basic training in Hayward for two weeks, I was teamed with Darrel } for a couple of weeks for WDS training and then left to install a dozen } instruments through out the midwest because there had been no one available } to do the installations for the previous two years or more. Forgive me if I } am a little touchy on this subject, but I had to go into many situations } where ETEC had simply been unable to completely install instruments that had } been delivered years before. I realize that I entered ETEC on the cusp of } their acquisition by Perkin-Elmer, but I was alone in bringing much of the } midwest territory into compliance with the contractual responsibilities that } ETEC undertook. } } I can only hope that this group will forgive my self-indulgence, but I have } to point out that Earl has done nothing to refute the suggestions that I } made. They still stand on their own. Earl's clarifications suggest that } Ken Converse knows what he is doing, a subject that I never refuted. On the } contrary, I have referred Ken to a number of people who have contacted me } for service over the years for a variety of instruments. In fact, I will be } packing up a Hitachi SEM in the next week that I have suggested Ken for the } installation and continuing service of. } } I stand by my comments on a fully-focusing WDS spectrometer being a } different beast requiring a careful and informed hand for proper maintenance } and operation. And I stand by my assertion that the ETEC WDS spectrometer } in particular is an extremely touchy and sensitive instrument to properly } calibrate. Once accomplished, however, the ETEC Autoscan mated with the } ETEC WDS is a very effective and stable electron microprobe system, } something I can attest to with my own personal instrument. } } Allen R. Sampson } Advanced Research Systems } 317 North 4th. Street } St. Charles, IL 60174 } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com } } -----Original Message----- } } From: "earlw-at-pacbell.net"-at-sparc5.microscopy.com } {"earlw-at-pacbell.net"-at-sparc5.microscopy.com} } To: Allen R. Sampson {ars-at-sem.com} } Cc: amenex-at-amenex.com {amenex-at-amenex.com} ; Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } Date: Friday, July 16, 1999 7:26 PM } Subject: Re: Haaalp ! How can I improve oxygen dot maps in microprobe ? } } } } } Dear Allen, } } } } You were one of several factory trained technicians: I was one, Ken was } another, } } Hank Bebe was also trained. I was trained by "Tung Tsu" who trained Darrell } } Jackson. Ask Darrell how many spectrometers he screwed up before being } trained } } by Tung Tsu. } } } } Earl } } } } "Allen R. Sampson" wrote: } } } } } } } } For the best and most trained of ETEC technicians, the alignment of the } WDS } } } crystals is a daunting task. The TAP crystal is not a particularily } } } sensitive crystal as far as cleaning, a simple quick swabing with ethanol } } } should remove most contamination. However, the ETEC crystal mount has } four } } } mounting screws that are adjusted to control the tilt of the crystal in } two } } } dimensions, the crystal height and alignment with the Rowland circle as } well } } } as the curvature of the crystal. } } } } } } Ken is a very good technician on the ETECs and was my technical backup as } a } } } field technician. However, I was the only technician, that I know of, } that } } } was trained by the gentleman who set up the WDS systems in the factory. } I } } } know from setting up many systems that I would often spend an entire week } } } adjusting a single crystal for proper operation. These are fully } focussing } } } Johansson WDS optics that are capable of measurements to one ten } thousandth } } } of an Angstrom. However, the adjustment screws are very course and } touchy } } } to properly adjust. Added to this is the proper adjustment of the } detector } } } tape, a metal tape that both maintains the line of sight of the detector } to } } } the crystals as well as maintaining the proper detector alignment to the } } } Rowland circle. The ETEC WDS spectrometer also requires an accurate } } } alignment of the spectrometer housing to the sample chamber. } } } } } } More than likely, your spectrometer is in need of a full alignment that } may } } } well cost you thousands of dollars and a couple of weeks of work or more } to } } } accomplish. } } } } } } I can put you in touch with a gentleman who can provide you with new } } } crystals, contact me if needed, but more than likely you have need of a } } } complete overhaul of the spectrometer system. } } } } } } Allen R. Sampson } } } Advanced Research Systems } } } 317 North 4th. Street } } } St. Charles, IL 60174 } } } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com } } } } } } -----Original Message----- } } } } From: George Langford, Sc.D. {amenex-at-amenex.com} } } } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } } } Date: Thursday, July 15, 1999 1:09 PM } } } Subject: Haaalp ! How can I improve oxygen dot maps in microprobe ? } } } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 probing Microscopists ! } } } } } } } } Now that I've gotten past the hyperactive spam filter ... } } } } } } } } Amenex has a 1976-vintage ETEC Autoprobe SEM with three } } } } WDS spectrometers. Most of the instrument still functions } } } } acceptably (with the able assistance of Ken Converse) but } } } } we have noticed substantial deterioration of the oxygen } } } } dot maps in the last few years. The sodium maps come out } } } } OK; nitrogen has always been pretty much hopeless; but } } } } carbon works OK. } } } } } } } } We've been told that our TAP crystal may have deteriorated } } } } or become contaminated. What can we do about that ? Are } } } } there any adjustments or alignments that could be tweaked } } } } so as to pick up the oxygen radiation better ? } } } } } } } } Has anyone got a spare TAP crystal for sale or trade ? We } } } } have several spares of other crystals ... } } } } } } } } Any advice or suggestions would be greatly appreciated. } } } } } } } } Best regards, } } } } George Langford, Sc.D. } } } } amenex-at-amenex.com } } } } http://www.amenex.com/ } } } } } } } } } } } } } } } } } } } } Alan,
George said he has an ETEC Autoprobe, not an Autoscan with Autospec. The Autoprobe has 3 MAC spectrometers (also fully focussing and single crystal, single detector). I believe Hank hit it on the nose and Jim Nicholino makes great crystals.
You did not single-handedly square away the midwest. There were many of us who spent weeks at a time out there after our own areas were in pretty good shape, enough time to allow them to go to hell in a handbasket again so we'd have something to do when we got back home.
} Don't unsubscribe simply because the recent threads haven't been to your } liking - you might miss some great stuff! I agree 95% of the mailing list traffic is of little interest to=20 me. I automatically filter it into a mail folder with Eudora and=20 look for interesting subjects. After a while I file what is useful=20 and consign the rest to the bit bucket!
Please note new FAX number.
John Mansfield PhD CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Re: what isn't you area: Yes, sometimes just seeing something presented starts the wheels turning..... I have acquired my best obsessions and vices in this manner! Don't Un Sub, Just use the delete key after you have scanned through it!
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Allen R. Sampson 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 } } -----------------------------------------------------------------------. } } } } Glad to hear it, Earl. I had never heard from either Hank or Ken that they } } had been properly trained in WDS, and still have not. To the contrary, } } after I first came on board, received orientation in Danbury for a week and } } received basic training in Hayward for two weeks, I was teamed with Darrel } } for a couple of weeks for WDS training and then left to install a dozen } } instruments through out the midwest because there had been no one available } } to do the installations for the previous two years or more. Forgive me if I } } am a little touchy on this subject, but I had to go into many situations } } where ETEC had simply been unable to completely install instruments that had } } been delivered years before. I realize that I entered ETEC on the cusp of } } their acquisition by Perkin-Elmer, but I was alone in bringing much of the } } midwest territory into compliance with the contractual responsibilities that } } ETEC undertook. } } } } I can only hope that this group will forgive my self-indulgence, but I have } } to point out that Earl has done nothing to refute the suggestions that I } } made. They still stand on their own. Earl's clarifications suggest that } } Ken Converse knows what he is doing, a subject that I never refuted. On the } } contrary, I have referred Ken to a number of people who have contacted me } } for service over the years for a variety of instruments. In fact, I will be } } packing up a Hitachi SEM in the next week that I have suggested Ken for the } } installation and continuing service of. } } } } I stand by my comments on a fully-focusing WDS spectrometer being a } } different beast requiring a careful and informed hand for proper maintenance } } and operation. And I stand by my assertion that the ETEC WDS spectrometer } } in particular is an extremely touchy and sensitive instrument to properly } } calibrate. Once accomplished, however, the ETEC Autoscan mated with the } } ETEC WDS is a very effective and stable electron microprobe system, } } something I can attest to with my own personal instrument. } } } } Allen R. Sampson } } Advanced Research Systems } } 317 North 4th. Street } } St. Charles, IL 60174 } } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com } } } } -----Original Message----- } } } From: "earlw-at-pacbell.net"-at-sparc5.microscopy.com } } {"earlw-at-pacbell.net"-at-sparc5.microscopy.com} } } To: Allen R. Sampson {ars-at-sem.com} } } Cc: amenex-at-amenex.com {amenex-at-amenex.com} ; Microscopy-at-sparc5.microscopy.com } } {Microscopy-at-sparc5.microscopy.com} } } Date: Friday, July 16, 1999 7:26 PM } } Subject: Re: Haaalp ! How can I improve oxygen dot maps in microprobe ? } } } } } } } } Dear Allen, } } } } } } You were one of several factory trained technicians: I was one, Ken was } } another, } } } Hank Bebe was also trained. I was trained by "Tung Tsu" who trained Darrell } } } Jackson. Ask Darrell how many spectrometers he screwed up before being } } trained } } } by Tung Tsu. } } } } } } Earl } } } } } } "Allen R. Sampson" wrote: } } } } } } } } } } } For the best and most trained of ETEC technicians, the alignment of the } } WDS } } } } crystals is a daunting task. The TAP crystal is not a particularily } } } } sensitive crystal as far as cleaning, a simple quick swabing with ethanol } } } } should remove most contamination. However, the ETEC crystal mount has } } four } } } } mounting screws that are adjusted to control the tilt of the crystal in } } two } } } } dimensions, the crystal height and alignment with the Rowland circle as } } well } } } } as the curvature of the crystal. } } } } } } } } Ken is a very good technician on the ETECs and was my technical backup as } } a } } } } field technician. However, I was the only technician, that I know of, } } that } } } } was trained by the gentleman who set up the WDS systems in the factory. } } I } } } } know from setting up many systems that I would often spend an entire week } } } } adjusting a single crystal for proper operation. These are fully } } focussing } } } } Johansson WDS optics that are capable of measurements to one ten } } thousandth } } } } of an Angstrom. However, the adjustment screws are very course and } } touchy } } } } to properly adjust. Added to this is the proper adjustment of the } } detector } } } } tape, a metal tape that both maintains the line of sight of the detector } } to } } } } the crystals as well as maintaining the proper detector alignment to the } } } } Rowland circle. The ETEC WDS spectrometer also requires an accurate } } } } alignment of the spectrometer housing to the sample chamber. } } } } } } } } More than likely, your spectrometer is in need of a full alignment that } } may } } } } well cost you thousands of dollars and a couple of weeks of work or more } } to } } } } accomplish. } } } } } } } } I can put you in touch with a gentleman who can provide you with new } } } } crystals, contact me if needed, but more than likely you have need of a } } } } complete overhaul of the spectrometer system. } } } } } } } } Allen R. Sampson } } } } Advanced Research Systems } } } } 317 North 4th. Street } } } } St. Charles, IL 60174 } } } } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com } } } } } } } } -----Original Message----- } } } } } From: George Langford, Sc.D. {amenex-at-amenex.com} } } } } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } } } } Date: Thursday, July 15, 1999 1:09 PM } } } } Subject: Haaalp ! How can I improve oxygen dot maps in microprobe ? } } } } } } } } } ------------------------------------------------------------------------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 probing Microscopists ! } } } } } } } } } } Now that I've gotten past the hyperactive spam filter ... } } } } } } } } } } Amenex has a 1976-vintage ETEC Autoprobe SEM with three } } } } } WDS spectrometers. Most of the instrument still functions } } } } } acceptably (with the able assistance of Ken Converse) but } } } } } we have noticed substantial deterioration of the oxygen } } } } } dot maps in the last few years. The sodium maps come out } } } } } OK; nitrogen has always been pretty much hopeless; but } } } } } carbon works OK. } } } } } } } } } } We've been told that our TAP crystal may have deteriorated } } } } } or become contaminated. What can we do about that ? Are } } } } } there any adjustments or alignments that could be tweaked } } } } } so as to pick up the oxygen radiation better ? } } } } } } } } } } Has anyone got a spare TAP crystal for sale or trade ? We } } } } } have several spares of other crystals ... } } } } } } } } } } Any advice or suggestions would be greatly appreciated. } } } } } } } } } } Best regards, } } } } } George Langford, Sc.D. } } } } } amenex-at-amenex.com } } } } } http://www.amenex.com/ } } } } } } } } } } } } } } } } } } } } } } } } } } } } Alan, } } George said he has an ETEC Autoprobe, not an Autoscan with Autospec. } The Autoprobe has 3 MAC spectrometers (also fully focussing and single } crystal, single detector). I believe Hank hit it on the nose and Jim } Nicholino makes great crystals. } } You did not single-handedly square away the midwest. There were many of } us who spent weeks at a time out there after our own areas were in } pretty good shape, enough time to allow them to go to hell in a } handbasket again so we'd have something to do when we got back home. } } Ken Converse } owner } Quality Images
} Hello Greg! } I feel you haven't understand my intentions. } I found acces to this listserver by way of the archives you named. I got } useful hints from the archives and decided to participate in the } listserver. I am not critizising the support that the MSA Listserver } provides, sure it is the most advanced forum in special microscopy ! } But I'm referring to the shift of themes and topics discussed in the } listserver and the obviously ignorance of basic themes ( embedding, } proceedings). Costs of new imaging and archiving possibilities, offers } of postdoctoral's and used devices seem to be overwhelming in the recent } past.
No, Michael, I think it's you who doesn't quite understand the intentions of the list. It's a forum for the exchange of ideas, not a publication mechanism. Topics discussed change from time to time, as you point out, but the initiative is usually a question, not someone's desire to tell something to the world. Imaging and archiving, post-docs, and used equipment may not interest you, but they obviously do interest many, as evidenced by the volume of correspondance. So the thing is self-regulating in that if there's no interest, a thread stops.
I have to admit to being a bit incensed by your words "the obviously ignorance of basic themes ( embedding, proceedings)" .
This is not a place for the publication or the acquisition of basic information, there are heaps of texts for that. But if you have a SPECIFIC question, go ahead and ask it, it'll probably get answered.
Just quit this "If you guys don't write about what I want, I'm going to unsubscribe! So there!" stuff.
I'd rather read a posting on imaging, vacuum problems, or even ETEC service training, any day than be subjected to your petulance.
Unsubscribe! Unsubscribe! Do it now!
cheers
rtch
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
a couple of years ago I purchased DTSA version 2.5.1 and have used it only occassionally for qualitative studies. I now wish to make full use of it's quantitative analysis functions for EDX spectra from TEM specimens (Cliff-Lorimer analysis of materials specimens). I have a few questions that I'm hoping you guys can help me with.
1) is there a more recent update of DTSA? I note from the documentation that some functions were not fully implimented in V2.5.1;
2) is DTSA still being developed?
3) is there a plug-in for converting Link ISIS EDX spectrum files into DTSA format?
3) is there a plug-in for converting EMiSpec ES Vision EDX spectrum files into DTSA format? I know this is a long shot but I thought I'd ask anyway.
4) if these plug-in are not currently available, what do I need to write my own (assuming I can get details of the file formats)?
Any help with these questions, or suggestions regarding the use of DTSA for Cliff-lorimer analysis would be most welcome. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
Please excuse me if you received this posting earlier. I received a message from Mail Delivery Subsystem {MAILER-DAEMON-at-Sparc5.Microscopy.Com} with the following message in the subject line: "Returned mail: Bad usage". I assume the anti-spam filter was offended by something in my posting, possibly the subject line, so I'll try again:
a couple of years ago I purchased DTSA version 2.5.1 and have used it only occassionally for qualitative studies. I now wish to make full use of it's quantitative analysis functions for EDX spectra from TEM specimens (Cliff-Lorimer analysis of materials specimens). I have a few questions that I'm hoping you guys can help me with.
1) is there a more recent update of DTSA? I note from the documentation that some functions were not fully implimented in V2.5.1;
2) is DTSA still being developed?
3) is there a plug-in for converting Link ISIS EDX spectrum files into DTSA format?
3) is there a plug-in for converting EMiSpec ES Vision EDX spectrum files into DTSA format? I know this is a long shot but I thought I'd ask anyway.
4) if these plug-in are not currently available, what do I need to write my own (assuming I can get details of the file formats)?
Any help with these questions, or suggestions regarding the use of DTSA for Cliff-lorimer analysis would be most welcome. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} Hello Greg! } I feel you haven't understand my intentions. } I found acces to this listserver by way of the archives you named. I got } useful hints from the archives and decided to participate in the } listserver. I am not critizising the support that the MSA Listserver } provides, sure it is the most advanced forum in special microscopy ! } But I'm referring to the shift of themes and topics discussed in the } listserver and the obviously ignorance of basic themes ( embedding, } proceedings). Costs of new imaging and archiving possibilities, offers } of postdoctoral's and used devices seem to be overwhelming in the recent } past. } }
The thing is Michael - this list was not devised by you nor is it run by you. It is an invaluable forum for the sharing of thoughts and for questions and for just straight contact with colleagues. It takes on a life of its own in terms of content. Subjects that are of more general interest will clearly predominate. If there is little interest in a subject notice how quickly it fizzles out.
Look - it is soooo easy to hit the delete button if you are not interested in a subject. If you do have subjects you are personally really interested in then post some good solid questions (after first exhausting on-hand texts) and for sure you will receive answers.
And unltimately if you don't find enough value in the List then it is easy to unsubscribe.
Believe me there is a golden fountain of experienced, knowledgeable people on this list. Don't belittle it.
Dear all, Please don't flame someone just because they say things on the server you don't like (for instance a recent discussion about microprobe service, and comments about someone suggesting that he may unsubscribe due to lack of response to queries in his area). If you really think someone is out of line, please talk to Nestor instead.
Best wishes to all
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics Chalmers University of Technology S-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 FAX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com Research Group Homepage: http://fy.chalmers.se/microscopy/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Dear members of the list, it wasn=B4t my intention to offend anybody with my thoughts about the "shift of themes" on this listserver. I feel that many from you are angry and irritated by my thoughts.
First, I was content with your cooperation. Every question I asked to you was replied satisfactorily and I got a lot of useful hints which lead to advances in my work. In fact I was very happy about the great response. I wanted to thank you and not to say "If you guys don't write about what I want, I'm going to unsubscribe!".
Second, I just noticed a "shift of themes" but obviously I made me an "outsider" by these statements.
Third, I mentioned the "ignorance of basic themes". This was overaccaregated. But it is true that sometimes a "newcomer" has a basic question which remains unanswered. I remember a case of someone who wanted to do in-situ on thin sections for electron microscopy. At least I replied. This made me feel, I don=B4t know, alone. It was the first time that I had the possibility to answer someone but nobody else engaged discussion and this topic left "dead". A question about DAB and immunogold combination remained unanswered up to now. As well as another question dealing with LR-White and immunogold (posted by Sonya Cawsey). There are more examples in the recent past. These are topics which are -in fact- basics. Maybe I=B4m wrong ...
I fell sorry about offending you with my mails. As you see I=B4m still subscribed and I will do so further. I hope to be able to do my part to keep the mailing list attractive even for newcomers as I am with many questions and few answers. I hope you all can calm down now and I you will accept me as a staying member of the list in the future. Sorry.
Dear all, Please don't flame someone just because they say things on the server you don't like (for instance a recent discussion about microprobe service, and comments about someone suggesting that he may unsubscribe due to lack of response to queries in his area). If you really think someone is out of line, please talk to Nestor instead.
Best wishes to all
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics Chalmers University of Technology S-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 FAX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com Research Group Homepage: http://fy.chalmers.se/microscopy/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Dear members of the list, it wasn=B4t my intention to offend anybody with my thoughts about the "shift of themes" on this listserver. I feel that many from you are angry and irritated by my thoughts.
First, I was content with your cooperation. Every question I asked to you was replied satisfactorily and I got a lot of useful hints which lead to advances in my work. In fact I was very happy about the great response. I wanted to thank you and not to say "If you guys don't write
about what I want, I'm going to unsubscribe!".
Second, I just noticed a "shift of themes" but obviously I made me an "outsider" by these statements.
Third, I mentioned the "ignorance of basic themes". This was overaccaregated. But it is true that sometimes a "newcomer" has a basic question which remains unanswered. I remember a case of someone who wanted to do in-situ on thin sections for electron microscopy. At least I replied. This made me feel, I don=B4t know, alone. It was the first time that I had the possibility to answer someone but nobody else engaged discussion and this topic left "dead". A
question about DAB and immunogold combination remained unanswered up to now. As well as another question dealing with LR-White and immunogold (posted by Sonya Cawsey). There are more examples in the recent past. These are topics which are -in fact- basics. Maybe I=B4m wrong ...
I fell sorry about offending you with my mails. As you see I=B4m still subscribed and I will do so further. I hope to be able to do my part to keep the mailing list attractive even for newcomers as I am with many questions and few answers. I hope you all can calm down now and I you will accept me as a staying member of the list in the future. Sorry.
Dear members of the list, it wasn=B4t my intention to offend anybody with my thoughts about the "shift of themes" on this listserver. I feel that many from you are angry and irritated by my thoughts. I didn=B4t want to make enemies.
First, I was content with your cooperation. Every question I asked to you was replied satisfactorily and I got a lot of useful hints which lead to advances in my work. In fact I was very happy about the great response. I wanted to thank you and not to say "If you guys don't write
about what I want, I'm going to unsubscribe!".
Second, I just noticed a "shift of themes" but obviously I made me an "outsider" by these statements.
Third, I mentioned the "ignorance of basic themes". This was overaccaregated. But it is true that sometimes a "newcomer" has a basic question which remains unanswered. I remember a case of someone who wanted to do in-situ on thin sections for electron microscopy. At least I replied. This made me feel, I don=B4t know, alone. It was the first time that I had the possibility to answer someone but nobody else engaged discussion and this topic left "dead". A
question about DAB and immunogold combination remained unanswered up to now. As well as another question dealing with LR-White and immunogold (posted by Sonya Cawsey). There are more examples in the recent past. These are topics which are -in fact- basics. Maybe I=B4m wrong ...
I fell sorry about offending you with my mails. As you see I=B4m still subscribed and I will do so further. I hope to be able to do my part to keep the mailing list attractive even for newcomers as I am with many questions and few answers. I hope you all can calm down now and I you will accept me as a staying member of the list in the future. Sorry.
I have found the cause of the Email bounce. It was not do to anyone's postings but due to a edit I was doing in the software. Don't hassel anyone else ..
Dear Frank In reference to your e-mail dated 21/7/99.
1) the smallest particle should be the same as you observe on the SEM screen. When taking images and making measurements on features in a package such as IMQuant, you should use the highest magnification possible for small features to minimise any feature detection and measurement errors. e.g. if you use a low mag and the features of interest only occupy a few pixels in your image then area and shape measurments could have large errors. If you raise the mag then the same features occupy tens or hundreds of pixels then the measurements will be much more accurate.
2) There is no compensation within Imquant. This is a classic stereological problem and is described in a number of books. One book you could try is "Stereological methods" Practical methods for biological Morphometry by Ewald R Weibel published by Academic Press Limited chapter 5.
Regards
Oxford Instruments Microanalysis Ltd. Customer Support Group ----------------- } From: frank.sarrazit-at-AVESTASHEFFIELD To: MAG-TSG Hotline
Hi I have a couple of questions related to the IMquant software and would be grateful if you could help. 1/ What's the resolution /smallest particle which can be accurately detected? Is it different from that of the microscope? 2/ we are looking at 2-D sections of steel blades and have successfully used IMQuant to characterise carbide particles (area etc ). However, because these are 2-D sections, diameters and carbides are understated and I was wondering whether a mathematical way of compensating for this was available within imquant? Thanks for your help Franck
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(InterLock SMTP Gateway 3.0 for Microscopy-at-sparc5.microscopy.com); Mon, 26 Jul 1999 08:06:23 -0500 Received: by gate.mmd.com (Protected-side Proxy Mail Agent-1); Mon, 26 Jul 1999 08:06:23 -0500 Message-Id: {A16C87C13FB5D1118BD000A0C9AF993601172766-at-brwsmxsusr04.brw.hmrag.com}
Dear Sally,
Spreading DNA is a very tedious procedure. I am not familiar with the spontaneous adsorption technique, but I know the spreading technique has always worked for me and I'm including it here. But first, let me ask you several questions. What buffer are you using and at what pH? This is very important. A more alkaline solution has always worked for me (see below). Second, what is the purpose of ammonium acetate? I have never used this and achieved excellent results. Third, what is your hypophase solution? This is very important since this serves as a matrix. Look at my procedure. It is a very modified one and I cannot locate the original source right now.
Spreading solution: Working Solution:
1 M Tris buffer - pH 8.5 + 0.1M EDTA 10ul 99% formamide - highly purified molecular bio. grade 50ul
DNA - (50ug/ml stock solution) 2ul ddH2O - 33ul cytochrome C 5ul always add last
Add in order given. Make sure that any glass used, including slides must be soaked in HCl/Chromic acid solution for 2 days prior to use. Also, as you know,everything must be DNAase free.
Hypophase - floating solution.
ddH2O 79ml 1M Tris (above) + 0.1M EDTA pH 8.5 1ml formamide 20ml The procedure is the slide spreading technique. The DNA is collected onto carbon-coated grids 400m. The next step was to stain in 0.05M uranyl acetate in 95% ethanol and 0.1M HCl.
Please give me a call if you have any questions and I would be more than happy to explain further the technique. Blessings................................................................... Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Hoechst Marion Roussel, Inc. 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-hmrag.com
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Michael, I think you underestimate the people who participate on the list.=20 Maybe you did not get an answer because no-one had a good one for you. It is possible you know that what you ask may be something for which many of us do not have good answers. I have very little idea of who is a senior or=
a junior researcher on the list. Sure a few names pop out but very few.=20 I am a biologist and many of the participants on the list are in materials. I do not get upset that they may send more than their fair shar= e of the messages. I just delete those I am not interested in and go on. The same is with any specific topic that does not interest me. If even one person is interested than it has a place on the list. =20 I do not consider myself a "junior" since I have been in this business for over 30 years. However, I have more than once poised a questi= on and not gotten any answers. Chalk that up to the fact that there are still lots of unexplained situations or unsolved questions combined with very busy people who may not want to take time to just speculate or repeat information that can be found in any basic text on electron microscopic technique. Even one good answer is well worth your time poising questions and most of the good tips I have received have not been to questions I have poised but to ones others have, which have been answered by many people mor= e knowledgeable than I. Stay on the list and read questions and answers which=
are in your very general area of interest and I guarantee that you will get more out of the list than you will put in. In no time I think you will=
realize that you are not a "junior" but an equal in the eyes, or fingers" of those submitting the to list.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University =20 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Dear colleagues Maybe I will unsubscribe soon ... I had many questions, but seldom an answer. That=B4s because I=B4m one of the "juniors" . Thanks especially to Hildy Crowley and Charles Garber, they are very active in this forum and had good hints regarding embedding for immuno-electron-microscopy (e.g. LR-White). Now, I=B4m doing quite good work in this special discipline and its possible that you will notice it one day. Recently, themes have changed in this listserver. Costs of digital imaging archives, salaries, used devices to sell or search and special advertisements, job offers seem to be the overwhelming topics nowadays. I=B4ve noticed that "greenhorn questions" remained unanswered in the first half of the year ... As I am resident in Europe, offers of used devices and so on don=B4t interest me that much as the most of you all (in the U.S.). Thus, many messages from the listserver are not so interesting for me especially. Dear colleagues, why don=B4t you discuss more about techniques and methods? I am young and want to gain experience. Sure, digital archives of EM-photographs are now high-sophisticated but my intsitution won=B4t afford it in the near future. Eyeryone of us has the fear to talk about unpublished results or datas, but I feel that electron-microscopic techniques are so complex that they need a wide cooperation.
Thank you for your support. I will return to the listserver, that=B4s sure
Michael Reiner, cand. med Department of Anatomy I University of Colgne Germany Joseph-Stelzmann-Str. 9 50931 Koeln/Cologne Germany e-mail: a2811111-at-smail.uni-koeln.de Tel: +49-221-478-5519 Fax: +49-221-478-6411
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by post.its.mcw.edu (8.8.8/8.8.8) with ESMTP id JAA01178 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 26 Jul 1999 09:16:53 -0500 (CDT) Message-ID: {379C708E.38A73978-at-mcw.edu}
Hi, All -
I want to put a service contract to my Reichert-Jung Ultracut. Can any of you provide some information about it? I would like to know who provides such services in the US.
Gang Ning EM Facility Medical College of Wisconsin
Just a reminder, abstracts and early registrations are due August 2nd for the 5th California Microscopy Colloquium organized by the Northern California Society for Microscopy and The California State Universities and to be held at San Francisco State University on October 2nd..
Our web site (http://online.sfsu.edu/~camicro/) is ready to accept both your early registration and abstracts.
Also, please forward this message to others (especially students) who you think might be interested but might not yet be included on our email list.
Thank you and looking forward to seeing you in October.
Thank you to everyone who responded to my enquiry about bolt-on digital camera systems for TEM.
It seems that the resolution of digital systems still doesn't quite match that of conventional film, but image contrast is definitely not a problem, and the convenience is definitely quite tempting!
I reckon better cameras with more pixels should be just round the corner if the technology continues to improve at the current pace.....
Amanda EM Unit St George's Hospital Medical School awilson-at-sghms.ac.uk
On Mon, 26 Jul 1999 DUNNTEM-at-aol.com-at-Sparc5.Microscopy.Com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Michael Reiner wrote:- } } } } Hello Greg! } } I feel you haven't understand my intentions. } } I found acces to this listserver by way of the archives you named. I got } } useful hints from the archives and decided to participate in the } } listserver. I am not critizising the support that the MSA Listserver } } provides, sure it is the most advanced forum in special microscopy ! } } But I'm referring to the shift of themes and topics discussed in the } } listserver and the obviously ignorance of basic themes ( embedding, } } proceedings). Costs of new imaging and archiving possibilities, offers } } of postdoctoral's and used devices seem to be overwhelming in the recent } } past. } } } } } The thing is Michael - this list was not devised by you nor is it run by you. } It is an invaluable forum for the sharing of thoughts and for questions and } for just straight contact with colleagues. It takes on a life of its own in } terms of content. Subjects that are of more general interest will clearly } predominate. If there is little interest in a subject notice how quickly it } fizzles out. } } Look - it is soooo easy to hit the delete button if you are not interested in } a subject. If you do have subjects you are personally really interested in } then post some good solid questions (after first exhausting on-hand texts) } and for sure you will receive answers. } } And unltimately if you don't find enough value in the List then it is easy to } unsubscribe. } } Believe me there is a golden fountain of experienced, knowledgeable people on } this list. Don't belittle it. } } Ted Dunn } Maui, Hawaii } } } Micheal,
Reread the last sentence above. Then please stop complaining or quit this most valuable service. Hildegard H. Crowley University of Denver
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Dear members of the list, } it wasn=B4t my intention to offend anybody with my thoughts about the } "shift of themes" on this listserver. } I feel that many from you are angry and irritated by my thoughts. } =20 } First, I was content with your cooperation. Every question I asked to } you was replied satisfactorily and I got a lot of useful hints which } lead to advances in my work. In fact I was very happy about the great } response. I wanted to thank you and not to say "If you guys don't write } =20 } about what I want, I'm going to unsubscribe!". } =20 } Second, I just noticed a "shift of themes" but obviously I made me an } "outsider" by these statements. } =20 } Third, I mentioned the "ignorance of basic themes". This was } overaccaregated. But it is true that sometimes a "newcomer" has a basic } question which remains unanswered. } I remember a case of someone who wanted to do in-situ on thin sections } for electron microscopy. At least I replied. This made me feel, I don=B4t } know, alone. It was the first time that I had the possibility to answer } someone but nobody else engaged discussion and this topic left "dead". A } =20 } question about DAB and immunogold combination remained unanswered up to } now. As well as another question dealing with LR-White and immunogold } (posted by Sonya Cawsey). There are more examples in the recent past. } These are topics which are -in fact- basics. } Maybe I=B4m wrong ... } =20 } I fell sorry about offending you with my mails. } As you see I=B4m still subscribed and I will do so further. I hope to be } able to do my part to keep the mailing list attractive even for } newcomers as I am with many questions and few answers. } I hope you all can calm down now and I you will accept me as a staying } member of the list in the future. } Sorry. } =20 } Yours sincerly, } Michael Reiner } =20 } =20 } =20 } =20 } =20 } =20 Hi, Sorry I took offense as to what I considered an attack on one of my best sources to be able to do good work. Regarding the "no answers" to "basic questions". I could spend my entire day on the List Server writing answers to basic questions, which, in most cases are not at all basic! DAB-TEM is particularly involved and difficult. There simply is not the time to write a whole textbook chapter on a basic question. Wish I could! Bye, Hildy Crowley
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Colleagues... } } I have found the cause of the Email bounce. It was not do to anyone's } postings but due to a edit I was doing in the software. Don't hassel } anyone else .. } } My fault entirely.... } } Nestor } Your Friendly Neighborhood SysOp } } } } Hi, You can make as many mistakes as you like! We are so grateful for what you are doing. You can bounce things or edit or whatever! We are so happy to have the List Server that next to nothing bothers us! See you, Hildy Crowley
I'd just like to say that I have received very valuable advice from list members. It is important to note that with this list, when you hit "Reply", it goe= s to the sender, not the whole list. (A good idea, with all the "vacation email response" programs people have= !) *** So I received many messages off-list.***
Some questions may not get responses because they have no answer. Who can say why my immunolabeling of LR White LM sections didn't work (mainly with DAB, though I did try a gold-conjugated secondary to see how that would work). I've done everything that the books, articles, and very kind list members have suggested. I've tried new primary antibodies; longer incubation; higher and lower titres; etching with ethanol, hot citrate buffer and sodium methoxide; lower polymerization temperature. Sometimes stuff just doesn't work out. I'm switching to cryosectioning (which might mean I'll have to kill more fish, but at least I'll learn a new technique).
Anyway, I'm a junior and I'm pretty happy with the list. There's no censorship of posts, which I have seen on some email lists. There's free and open discussion of whatever the list members want to bring up. Yeah!
Michael Reiner wrote:
} question about DAB and immunogold combination remained unanswered up to } now. As well as another question dealing with LR-White and immunogold } (posted by Sonya Cawsey). There are more examples in the recent past. } These are topics which are -in fact- basics. } Maybe I=B4m wrong ... }
... -- Sonia Cawsey McGowan Copley Library, University of San Diego email: scawsey-at-acusd.edu home page: http://www.acusd.edu/~scawsey
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Dear All,
In many respects, I would agree with Michael's point that although = subjects are introduced on the list, often few replies re-appear. It = would be good to follow discussions on particular topics as they are = brought up.
However, as a guilty party, I know that once a topic is brought up, it is = usually discussed further off-line. Occasionally we then re-post a = summary of replies, but this is not often. Perhaps a little more on-line = discussion would be useful.
I enjoy it most when there are different points of view being expressed.
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
I am doing some work involving image acquisition of a sample which is driven by stepper motors. The 1" sample is viewed from a microscope which has a field of view of 0.5mm and is equipped with a CCD camera. Although the sample is initially set up to be well focused within the microscope, when it is moved to another location, the image can get seriously out of focus because of the sample surface roughness, limited depth of field of the microscope, and etc. I can move the sample up or down (closer to or further away from the microscope) by stepper motor so that the viewed area is still in focus. However, I would prefer an automation to manual adjustment. Anyone knows a tool or software that can tell if the image is focused or not?
Transmission Electron Microscopist Position in DSE
The principal mission of the Department of Drug Safety Evaluation (DSE) is to define the toxic potential of chemicals to the extent necessary and desirable for assessing risk to man under intended use conditions. The Pathology Department is an integral part of DSE providing expert consultation, histological and ultrastructural preparation of non-human tissues and interpretation of changes to advance new candidate development. You will perform routine transmission electron microscopy, including specimen acquisition, tissue processing, ultramicrotomy, operation of Hitachi 7100, and darkroom procedures. In addition, you will be responsible for projects requiring methods development in areas such as, autoradiography, immunocytochemistry, freeze substitution, cryoultramicrotomy, freeze drying, and correlative light, transmission, and confocal microscopy. You will provide technical and scientific support for Drug Safety Evaluation and Discovery Research projects encompassing diversified therapeutic areas ( e.g., Cardiovascular, Cancer, CNS, Animal Health, etc.).
A minimum of a BS and 2 years of relevant experience are required. Two or more years' experience with low temperature immuno-EM is preferred.
Pfizer is proud to offer competitive salaries, exceptional benefits, tremendous opportunities for learning and advancement and the rewards of living only hours from Boston, Providence and New York City on the beautiful Connecticut seashore. Please send your resume in confidence to:Pfizer Inc, Central Research, Job Code: XXXXXXNEW, c/o Aon Consulting, P.O. Box 25, Findlay, OH 45839, or e-mail to: _ HYPERLINK mailto:Pfizer-at-aon-hros.com __Pfizer-at-aon-hros.com_.Pfizer offers a workplace rich with diversity and potential - an Equal Opportunity Employer.
I am trained to operate a scanning electron microscope but I do not hold a certificate of any kind. Is certification useful in finding employment? If so, where would I go to get information on becoming certified?
Just a reminder, abstracts and early registrations are due August 2nd.
Our web site (http://online.sfsu.edu/~camicro/) is ready to accept both your early registration and abstracts.
Also, please forward this message to others (especially students) who you think might be interested but may not yet be included on our email list.
Thank you and looking forward to seeing you in October.
Gregory A. Antipa Department of Biology San Francisco State University San Francisco CA 94132 Office/Lab (415) 338-2951 Email antipa-at-sfsu.edu FAX (415) 338-2295
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Re} support of Reichert-Jung ultramicrotomes:
We had good service from TEKNET on the east coast (1-800 835 6386). I am = not sure they support Wisconsin though.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
by email.nist.gov (8.9.3/8.9.3) with ESMTP id RAA22504; Mon, 26 Jul 1999 17:06:51 -0400 (EDT) Sender: js0b-at-email.nist.gov Message-ID: {379CCD00.3B5E67E2-at-nist.gov}
Mark Blackford wrote: } } Dear All, } } a couple of years ago I purchased DTSA version 2.5.1 and have used it only } occassionally for qualitative studies. I now wish to make full use of it's } quantitative analysis functions for EDX spectra from TEM specimens } (Cliff-Lorimer analysis of materials specimens). I have a few questions } that I'm hoping you guys can help me with. } } 1) is there a more recent update of DTSA? I note from the documentation } that some functions were not fully implimented in V2.5.1; } } 2) is DTSA still being developed? } } 3) is there a plug-in for converting Link ISIS EDX spectrum files into DTSA } format? } } 3) is there a plug-in for converting EMiSpec ES Vision EDX spectrum files } into DTSA format? I know this is a long shot but I thought I'd ask anyway. } } 4) if these plug-in are not currently available, what do I need to write my } own (assuming I can get details of the file formats)? } } Any help with these questions, or suggestions regarding the use of DTSA for } Cliff-lorimer analysis would be most welcome. Cheers, } } Mark Blackford } TEM Group } Materials Division, ANSTO } PMB 1, } Menai, N.S.W. } Australia } 2234
Mark,
} 1) is there a more recent update of DTSA? I note from the documentation } that some functions were not fully implimented in V2.5.1;
The most recent "production" version of DTSA is 2.6.1, although this may not have been approved for release to the external website yet (http://micro.nist.gov/DTSA/dtsa.html).
This version includes substantial changes from 2.5.x, at least internally. Many of the data structures have been cleaned up, and some code has been modified (e.g. now it is much easier to loop over x-ray lines). Many of these changes, however, are not visible to the user. Unfortunately, many functions are still not implemented.
} 2) is DTSA still being developed?
Yes.
} 3) is there a plug-in for converting Link ISIS EDX spectrum files into DTSA } format? } 3) is there a plug-in for converting EMiSpec ES Vision EDX spectrum files } into DTSA format? I know this is a long shot but I thought I'd ask anyway. } 4) if these plug-in are not currently available, what do I need to write my } own (assuming I can get details of the file formats)? } } Any help with these questions, or suggestions regarding the use of DTSA for } Cliff-lorimer analysis would be most welcome. Cheers,
As far as I know, there is no plug-in for Link ISIS EDS spectra or EMiSPEC Vision spectral files. It would be straightforward to write these plug-ins, but I'm not sure that would be the right approach. A lot of time was invested in refining the MSA spectrum file format so it could act as a standard for exchanging data between acquisition/analysis applications. I believe ISIS can already export MSA files (they call it "EMSA/MAS file" in my version) which can be read by DTSA. As for EMiSPEC, I'd rather see an EMiSPEC-} MSA converter than a DTSA plug-in for EMiSPEC spectral files. I think a quick GUI-based tool (written in VB?, scripted in Vision itself using the COM objects exposed in their hierarchy?) would be useful for plucking out specific spectra (or a list of spectra) from the spectral sequences used in spectrum images and spectrum profiles.
If you decide you do want to write a DTSA plug-in, have a look at the source code for the existing plug-ins. There are working examples for reading and writing binary and ASCII files. The plug-ins were patterned after Adobe Photoshop plug-ins. The data details are different, of course, so the two are not compatible. For the input case, the basic idea is to write a completely independent chunk of code that parses the incoming spectral file and populates a data structure that the main program can read. This pre-defined area of memory is set up by DTSA when it is launched, when it scans for plug-ins and calls each one to see if it is functional. When the plug-in is called to read a file, DTSA passes control to the plug-in code, the plug-in reads the file and fills the data structure, the plug-in returns, and DTSA resumes execution where it left off, assuming the pre-defined memory area now contains data from the file. Thus, there are no parameters or data passed between DTSA and the plug-in. This makes it easier for you to write the plug-in using a different language than DTSA (e.g. C, or python) since you do not need to worry about the differences in calling convention. Also, you do not need to rebuild DTSA from source to write the plug-in (that's the point of a plug-in), but you may find it useful to look at the source for DTSA while writing. Agreement about the details of the (common) data structure shared between DTSA and all the plug-ins is assured by placing this data structure in an include file. All plug-ins should pull the data format from this include to prevent compatibility problems.
Hope this helps,
--John Henry
John Henry J. Scott NIST Microanalysis Research Group http://www-sims.nist.gov/Division/Contacts.html
Would you happen to know what solution would etch copper but not } stainless } } steel? I need this information to clean copper off a stainless steel fixture. } } thank you, } }
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Just a note to comment on DAB reactions for immunocytochemistry:
Apart from the fact that diamino-benzidine (DAB) is toxic, it is not = really a good visualization tool for immunocytochemistry. =
Firstly, it is non-quantitative. The amount of reaction product does not = depend on the number of antibodies binding to antigen, rather it is a = result of the reaction time between the HRP and the DAB (longer time =3D = more signal). It is an amplification reaction, not an improvement in = sensitivity. What we really want is a method that will give us the = location of an antigen AND an idea of the relative number of antigens = present.
Secondly, it is not a precise marker. With the enzyme reaction that = produces the precipitate comes the production of oxygen. This damages = intracellular organelles (if the antigen is located within the cell) and = causes reaction product to be distributed at sites other than where the = antigen is localized. This is also a result when the amplification of = reaction product occurs. =
Perhaps for these reasons there was little discussion on DAB problems. = Better to use the gold particles instead.
I get my information from papers published in the 1970's and will provide = the detailed references only if requested.
As for Michael Reiner, he is lucky enough to live in Germany, which is = well served by free, or low cost, instruction courses. To really get the = information you want (basic, detailed and served with a smile) I suggest = looking to FEBS or EMBL for help. They provide money for junior = microscopists to attend courses on these techniques.
PS: any idea why it takes my postings two days to appear? Sometimes they = don't even appear at all, which is worse!
Good luck.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Because of questions about my earlier comments on quantitative measurement of films in TEM, I thought I should provide clearer details. I want to offer the hope that labs without (expensive) CCD cameras can get quantitative data from film, perhaps for preliminary measurements or for student projects. Such techniques generally require more work than using a CCD camera, and they typically serve a different range of problems, but they are possible with a small investment!
The density D of an area of film that has received an exposure I to electrons in a TEM is given by
D = D(max)*(1-exp(-aI)),
where a is a constant. When D is much less than D(max), the exponential can be expanded and truncated to the first term, yielding
D = kE,
where k is a constant (Zeitler, 1992, Ultramicroscopy 46, 405). Experimental data showing the linearity of the film response for electrons is given in figure 4 of the paper cited. This paper notes that the linearity is valid for D(max) less than 1.5 for all commercially available TEM emulsions. It is assumed here that the base and fog density of the film d has been subtracted out. For practical work, it is useful to be explicit about the base and fog density, yielding the equation
D = kE + d.
To use this equation in a practical way, the specimen to be measured is placed on a holey film. When the image is made, D(max) is measured from the image of the hole on the film (because this is where the total beam current is recorded). D is measured from any desired point on the specimen. The base and fog density d is measured from the shadow created by the rim of the film holder, or anyplace else that the film isn't exposed to electrons. Because all of the measurements are made on the same piece of film, the degree of development or the age of the film are not relevant because there doesn't need to be any comparison to any other piece of film.
Films like this will typically have a density range from something slightly above zero to something slightly above one, a range that is easily measured with an analog densitometer. Because all of the measurement are made on the same piece of film with the same densitometer, the type of illumination or degree of collimation of the densitometer is not relevant.
There's more work to do if the film needs to be digitized, because typical scanners are often limited to 8 bits of black & white. Then it may be necessary to use techniques for dividing up the film dynamic range, calibrating scanners with a photographic step tablet (scanning the step tablet at the same time as the film!), and linearizing the response.
As noted above, these methods aren't as easy as getting a picture from a CCD camera. But I hope that people in smaller EM labs, especially students, can remember that they have the option to think quantitatively about their micrographs even in the absence of a large digital investment.
There is such tool and attachment. What microscope do you have ?
Please reply directly to:
emeylan-at-csi.com
Emile Meylan SERCO Technical Services, Inc.
----- Original Message ----- } From: Tong Wang {tong-at-jlab.org} To: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com} Sent: Monday, July 26, 1999 12:44 PM
G'day All,
We are looking to purchase a Wafering saw, and I need information. If = anyone can suggest models and/or suppliers and provide me with contact = numbers or emails, or any other info I will be most grateful. =20
If you have a second hand one that you want to get rid of, we are also = interested. =20
I look forward to hearing from you. =20
Thanks=20
George
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 1793 +61 3 9925 2205 Fax: +61 3 9925 5290 Email: george.theodossiou-at-rmit.edu.au
Hello Sally - Just in case Maria is following the method, but does not know "why ammonium acetate". I used that about 27 years ago doing "Kleinschmitt" DNA spreading. The ammonium acetate is volatile - is disappears without leaving crystals. So you have the correct osmotic pressure pulling on the DNA molecules but not too much rubbish left in the background. I think that there are other good applications for that salt in EM. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Monday, July 26, 1999 11:06 PM, Fazio-Zanakis, Maria, HMR/US [SMTP:Maria.Fazio-Zanakis-at-hmrag.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Sally, } } Spreading DNA is a very tedious procedure. I am not familiar with the } spontaneous adsorption technique, but I know the spreading technique has } always worked for me and I'm including it here. But first, let me ask you } several questions. What buffer are you using and at what pH? This is very } important. A more alkaline solution has always worked for me (see below). } Second, what is the purpose of ammonium acetate? I have never used this and } achieved excellent results. Third, what is your hypophase solution? This is } very important since this serves as a matrix. Look at my procedure. It is a } very modified one and I cannot locate the original source right now. } } Spreading solution: } Working Solution: } } 1 M Tris buffer - pH 8.5 + 0.1M EDTA } 10ul } 99% formamide - highly purified molecular bio. grade } 50ul } } DNA - (50ug/ml stock solution) } 2ul } ddH2O - } 33ul } cytochrome C } 5ul always add last } } Add in order given. Make sure that any glass used, including slides must } be soaked in HCl/Chromic acid solution for 2 days prior to use. Also, as you } know,everything must be DNAase free. } } Hypophase - floating solution. } } ddH2O } 79ml } 1M Tris (above) + 0.1M EDTA pH 8.5 } 1ml } formamide } 20ml } The procedure is the slide spreading technique. The DNA is collected onto } carbon-coated grids 400m. } The next step was to stain in 0.05M uranyl acetate in 95% ethanol and 0.1M } HCl. } } Please give me a call if you have any questions and I would be more than } happy to explain further the technique. } Blessings................................................................... } Maria } } Maria Fazio-Zanakis } Bioimaging and Molecular Histology } Hoechst Marion Roussel, Inc. } 1-908-231-3357 } Fax: 1-908-231-3962 } Email: Maria.Fazio-Zanakis-at-hmrag.com }
Dear Frank In reference to your e-mail dated 21/7/99.
1) the smallest particle should be the same as you observe on the SEM screen. When taking images and making measurements on features in a package such as IMQuant, you should use the highest magnification possible for small features to minimise any feature detection and measurement errors. e.g. if you use a low mag and the features of interest only occupy a few pixels in your image then area and shape measurments could have large errors. If you raise the mag then the same features occupy tens or hundreds of pixels then the measurements will be much more accurate.
2) There is no compensation within Imquant. This is a classic stereological problem and is described in a number of books. One book you could try is "Stereological methods" Practical methods for biological Morphometry by Ewald R Weibel published by Academic Press Limited chapter 5.
Regards
Oxford Instruments Microanalysis Ltd. Customer Support Group ----------------- } From: frank.sarrazit-at-AVESTASHEFFIELD To: MAG-TSG Hotline
Hi I have a couple of questions related to the IMquant software and would be grateful if you could help. 1/ What's the resolution /smallest particle which can be accurately detected? Is it different from that of the microscope? 2/ we are looking at 2-D sections of steel blades and have successfully used IMQuant to characterise carbide particles (area etc ). However, because these are 2-D sections, diameters and carbides are understated and I was wondering whether a mathematical way of compensating for this was available within imquant? Thanks for your help Franck
____________________________________________________________ Oxford Instruments Microanalysis, Halifax Road, High Wycombe,Bucks, HP12 3SE, UK Tel: +44 (0)1494 442255 Fax: +44 (0)1494 524129 http://www.oxford-instruments.com/ We have a 1M email size limit
Unless stated above to be non-confidential, this E-mail and any attachments are private and confidential and are for the addressee only and may not be used, copied or disclosed save to the addressee. If you have received this E-mail in error please notify us upon receipt and delete it from your records. Internet communications are not secure and Oxford Instruments is not responsible for their abuse by third parties nor for any alteration or corruption in transmission. ____________________________________________________________
Leica provides service for Reichert ultramicrotomes. For information from their office in Illinois, call 1-800-248-0223.
Best of luck,
Robyn
Robyn Rufner, Ph.D. Director, The Center for Microscopy and Image Analysis Ross Hall, Suite 406 Adjunct Associate Professor, Anatomy and Cell Biology THE GEORGE WASHINGTON UNIVERSITY 2300 Eye Street, N.W., 431 Ross Hall Washington, DC 20037-2337 Voice: (202) 994-2881 Fax: (202) 994-8885 Internet: anarrr-at-gwumc.edu
My apologies to those who hit a firewall while trying to download DTSA after my post yesterday. Sometimes the summer heat causes my IQ to drop into the single digits.
Try this URL: http://www.nist.gov/cstl/div837/837.02/MicroscopySoftware.html
} From there you can link to DTSA, MacLispix, and the NIST Monte Carlo pages. If you still have trouble accessing these URLs, please send me a private email and I'll try again.
-- John Henry
John Henry J. Scott NIST Microanalysis Research Group http://www-sims.nist.gov/Division/Contacts.html
I want to put a service contract to my Reichert-Jung Ultracut. Can any of you provide some information about it? I would like to know who provides such services in the US.
Gang Ning EM Facility Medical College of Wisconsin
One of my colleagues (who occasionally uses microscopy but is not a = microscopist) has asked me for my opinion . He was telling me about an = amazing demonstration that was given at his facility where a Richardson = RT-2 microscope was used to give results much better than a standard LM. = Apparently this was possible with some modifications of a LM, but the = exact mechanism for this vast improvement in image quality is not apparent = to me from the brief fact sheet which I have. I haven't heard of this = microscope before, nor have I seen it in action, but this question has = peaked my interest.
I would appreciate hearing from anyone who has this equipment, or has seen = a demonstration of it. If vendors are subscribed to this list and can = provide more information, I would appreciate that, too.=20
As always, thanks in advance.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
We have software that does exactly what you want. You can drive a stage, adjust the focus and finally montage the individual images into a large image with high resolution.
Please contact me through email for further details or call.
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
----- Original Message ----- } From: Tong Wang {tong-at-jlab.org} To: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com} Sent: Monday, July 26, 1999 12:44 PM
Dear All:
We are looking for a color CCD camera to capture true color images of plant samples. We would like to capture color images from fluorescently stained samples ( FITC, DAPI, Texas red), GFP as well as samples for bright-field/Transmitted light with traditional histology stains.. Currently we are considering the SPOT2 (Diagnostic Instruments), SenSys Color (Roper) and the color chilled C5810 ( Hamamatsu). Does anyone have any experience with these cameras? Any feedback will be appreciated. If anyone has any other recommended models of a good color CCD camera for microscopes other then the three cameras mentioned above please let me know.
Thanks in advance for your help
Sincerely,
Elison ***************************** Elison B. Blancaflor Plant Biology Division The Samuel Roberts Noble Foundation Ardmore, OK 73401
South Bay Technology, Inc. manufacturers diamond wafering saws, wire saws=
and also an acid saw. You can get general information on our saws via ou= r website at: www.southbaytech.com. In Australia, you can contact our exclusive distributor as follows:
Oxford Instruments P.O. Box 7 Pennant Hills NSW 2120 AUSTRALIA
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "George Theodossiou" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
G'day All,
We are looking to purchase a Wafering saw, and I need information. If anyone can suggest models and/or suppliers and provide me with contact numbers or emails, or any other info I will be most grateful. =
If you have a second hand one that you want to get rid of, we are also interested. =
I look forward to hearing from you. =
Thanks =
George
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 1793 +61 3 9925 2205 Fax: +61 3 9925 5290 Email: george.theodossiou-at-rmit.edu.au
We have Metamorph from Universal Imaging. It has an auto-focus function which works surprisingly well with fluorescent, brightfield and DIC images. We configurated it do so some montages of large tissue sample (sections of tumor) in combination with a scan-stage. We are very pleased with it. If you need more detail, you might contact them. The web page is: http: "//www.image1.com/".
Our scope has a cooled CCD, Zeiss axioplan II motoried microscope, and motorized XY stage.
Good luck!
Xuejun
PS: I have not commercial link with Universal Imaging. I am just a happy user.
On Mon, 26 Jul 1999, Tong Wang wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, everyone } } I am doing some work involving image acquisition of a sample which is } driven by stepper motors. The 1" sample is viewed from a microscope which } has a field of view of 0.5mm and is equipped with a CCD camera. Although } the sample is initially set up to be well focused within the microscope, } when it is moved to another location, the image can get seriously out of } focus because of the sample surface roughness, limited depth of field of } the microscope, and etc. I can move the sample up or down (closer to or } further away from the microscope) by stepper motor so that the viewed area } is still in focus. However, I would prefer an automation to manual } adjustment. Anyone knows a tool or software that can tell if the image is } focused or not? } } Any input is appreciated. } } Tong } } } } }
*********************************************************************** Xuejun Sun, Ph.D. Dept. of Experimental Oncology Cross Cancer Institute 11560 University Ave. Edmonton Alberta T6G 1Z2, Canada
Hey all, Once again, as I was getting frustrated with the clutter downstairs in the dungeon, I came across some items. I thought I'd offer them up gratis before pitching them.
Two Westinghouse projection lamps (DMX 500W 125V)
One and a partial box of EM filaments for possibly a Philips 300 (?) A guess by Mark Farmer as the boxes are labeled only with the part number and date #6101 and 7 Aug 1964. First one to provide the correct box color and ID for the fungus growing on them wins... ;)
Any takers, give me the address and I'll load em up.
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu
I can't seem to find the link for abstracts for M&M'99. I'd swear that these were already on thge web... does anyone know the URL? I'm looking at the M&M website and maybe the link is right in front of my nose, but I don't see it...
Thanks,
Brendan Foran, Ph.D. Materials Analysis Group / Internal Technical Support SEMATECH Austin, TX
-----Original Message----- } From: Foran, Brendan (S) Sent: Tuesday, July 27, 1999 11:16 AM To: Microscopy-at-sparc5.microscopy.com
Hello to you all!
I have been looking for a set of negative cassettes (30 plates), a cassette magazine box, and a cassette receiver box for a Hitachi-600 TEM. Please let me know if any of you or your friends has a H-600 TEM being taken down apart and wants to give away or trade these things.
Thanks in advance.
Gang Ning EM Facility Medical College of Wisconsin 414-456-8344
I think did do a demonstration on that microscope at the MSC in Guelph, Canada. It was very interesting as the manufacturer claims a total magnification of about 15,000X (this includes TV screen magnification) from a light microscope. All the information is proprietary - he would not tell me what kinds of modifications he used to make the LM do this kind of magnification but did say he was willing to visit places and do demos. At the meeting, he was affiliated with Leica. I saw some live cells from asparagus "juice" and could see organelles. I think it makes a bridge between conventional LM (which generally magnify to 100X) and TEM (without all the preparation time). It is interesting enough to look at. I can't tell you much more than that as the fact sheet is rather short. All I can tell you is that I did see it!
Lesley Bechtold
At 10:16 AM 7/27/99 -0400, Paula Allan-Wojtas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Santosh, In my Vander Voort Metallography book, several of the Cu etches are based on ammonium hydroxide and hydrogen peroxide. The stainless steel should be resistant to these. An example is: 25 ml NH4OH 25 ml water 25 - 50 ml H2O2 (3%) Use fresh, add peroxide last. At 05:23 PM 7/26/99 -0400, you wrote:
} } } Would you happen to know what solution would etch copper but not } } stainless } } } steel? I need this information to clean copper off a stainless steel } fixture. } } } thank you, } } } } } Santosh Kurinec } skkemc-at-rit.edu } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Santosh Kurinec asked how to remove copper from stainless steel without damaging the stainless. Then Chris Jeffree suggested ferric chloride ...
Chlorides in general are death to stainless steel - pitting or stress corrosion cracking can occur, especially if the exposure occurs at temperatures above about 150 F or if the stainless steel is being plastically deformed at the same time.
I would use concentrated nitric acid, which will remove the copper in an instant. However, some stainless steels are sensitive to nitric acid (that's the reason there's a Huey test in ASTM A262) so the exposure should not be for long. The attack takes place along inclusions on exposed sections that were transverse to the direction of prior mechanical working. It's also called, "end grain attack." Stainless steels with welds in them should also not be exposed to the concentrated nitric acid for long times.
This should be done under a hood, of course.
Nonsusceptible stainless steels are quickly passivated by the nitric acid, so they shouldn't even etch.
The same technique works on carbon steel items coated with copper or with copper parts embedded in them.
Best regards, George Langford, Sc.D. (Metallurgy) amenex-at-amenex.com http://www.amenex.com/
I have placed a Public Domain version of our Direct Methods code fs98 in the directory http://www.numis.nwu.edu/fs98
While this code is mainly for determining surface structures from 2D TED or x-ray diffraction data, it can also be used for phase-extension of HREM data or (in some cases) for structure determination just from TED data. (The later is a work in progress.) The code is designed for UNIX machines, but could be converted to work on a PC if someone wants to do this.
Contact me directly for specific questions/bugs (there will be some).
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
After rummaging a bit on the internet I found the following URL:
http://www.bio-microtech.com/products/rtm/
You can check out the PDF file there for more info.
We have NO connection to this company or the microscope, so this time I will not include my disclaimer. I just remembered that I had seen it on the web. (Ask me for image acquisition from this scope, though) ;-)
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Lesley S. Bechtold[SMTP:LSB-at-ARETHA.JAX.ORG] } Sent: Tuesday, July 27, 1999 11:52:12 AM } To: Paula Allan-Wojtas; microscopy-at-sparc5.microscopy.com } Subject: Re: LM- Richardson Real Time Microscope } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Paula and everyone,
I think did do a demonstration on that microscope at the MSC in Guelph, Canada. It was very interesting as the manufacturer claims a total magnification of about 15,000X (this includes TV screen magnification) from a light microscope. All the information is proprietary - he would not tell me what kinds of modifications he used to make the LM do this kind of magnification but did say he was willing to visit places and do demos. At the meeting, he was affiliated with Leica. I saw some live cells from asparagus "juice" and could see organelles. I think it makes a bridge between conventional LM (which generally magnify to 100X) and TEM (without all the preparation time). It is interesting enough to look at. I can't tell you much more than that as the fact sheet is rather short. All I can tell you is that I did see it!
Lesley Bechtold
At 10:16 AM 7/27/99 -0400, Paula Allan-Wojtas wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A new company, called Illumea, will be exhibiting at the upcoming M&M meeting. Although their product is telemicroscopy software, they have found some innovative solutions to just this problem. I'd suggest you contact Dr. Jack Zeineh (Zee-na) at Illumea with your question. He has developed proprietary algorithms for stage movement and focus control which may be applicable. I'll also cc with this response.
His contact info: Phone: 800-323-3203 Email: jzeineh-at-illumea.com website: www.illumea.com
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 03:44 PM 7/26/99 -0400, Tong Wang wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am trying to find a supplier of a NiF2 Single Crystal which I could cut down into a thin section. If any one has a lead on a company that supplies single crystals, it would be greatly appreciated.
Lance Delzeit
Lance Delzeit NASA Ames Research Center M/S 239-4 Moffett Field, CA 94035-1000 ph# 650-604-0236 fax 650-604-1088 ldelzeit-at-mail.arc.nasa.gov
} } One of my colleagues (who occasionally uses microscopy but is not a } } microscopist) has asked me for my opinion . He was telling me about an } amazing } demonstration that was given at his facility where a Richardson } RT-2 microscope } was used to give results much better than a standard LM. } Apparently this was } possible with some modifications of a LM, but the } exact mechanism for this vast } improvement in image quality is not } apparent to me from the brief fact sheet } which I have. I haven't heard of } this microscope before, nor have I seen it in } action, but this question } has peaked my interest. } } I would appreciate hearing from anyone who has this equipment, or has seen } a } demonstration of it. If vendors are subscribed to this list and can } provide } more information, I would appreciate that, too.
Hi, Paula,
The Richardson microscope is a clever new design (Canadian, like yourself) that was described in detail last March in Proc. RMS 34(1):311-316. If we're talking about the sme "Richardson", it hardly violates any of the laws of physics, but it's a simple, low cost (~$100 U.S.) design that will probably appeal to schools, field microscopists, and the "third world". I haven't had a chance to use one yet, but I'm looking forward to it; it may be good for Project MICRO.
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Don't know anything personally but I did find the following website with info about the microscope:
http://www.bio-microtech.com/products/rtm/
Paula Allan-Wojtas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, All, } } One of my colleagues (who occasionally uses microscopy but is not a microscopist) has asked me for my opinion . He was telling me about an amazing demonstration that was given at his facility where a Richardson RT-2 microscope was used to give results much better than a standard LM. Apparently this was possible with some modifications of a LM, but the exact mechanism for this vast improvement in image quality is not apparent to me from the brief fact sheet which I have. I haven't heard of this microscope before, nor have I seen it in action, but this question has peaked my interest. } } I would appreciate hearing from anyone who has this equipment, or has seen a demonstration of it. If vendors are subscribed to this list and can provide more information, I would appreciate that, too. } } As always, thanks in advance. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
From going to the site and looking at there reference, looks like the instrument is using IR to do the imaging.
The following reference's where listed;
1. Test Slides: Diatoms to Divisions - What are you looking at?, Part 1, Tim Richardson, (1998), Proceedings of the Roy. Microsc. Soc.,; March 1998, 33(1), pages 3-9.
This reference was on the site at URL: http://www.bio-microtech.com/info/articles/article1.htm
2. Infrared Light in the Microscope: History, Theory and Practical Aspects, Tim Richardson, (1997), Proceedings of the Roy. Microsc. Soc., December 1998, 32(4), pages 229-235.
3. Practical Infrared Tim Richardson, (1997), Proceedings of the Roy. Microsc. Soc., December 1997, 32(4), pages 236-242.
4. Do You, Your Slides and Your CCD Camera Need Sunglasses? Tim Richardson, (1997), Proceedings of the Roy. Microsc. Soc.,;June 1997, 32(2), pages
Dose any one know if the other reference's (2, 3 and 4) listed are some where on the net?
Best Regards
Joseph Passero jp-at-spacelab.net
Michael Bode wrote: } } } After rummaging a bit on the internet I found the following URL: } } http://www.bio-microtech.com/products/rtm/ } } You can check out the PDF file there for more info. } } We have NO connection to this company or the microscope, so this time I } will not include my disclaimer. I just remembered that I had seen it on } the web. (Ask me for image acquisition from this scope, though) ;-) } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } From: Lesley S. Bechtold[SMTP:LSB-at-ARETHA.JAX.ORG] } } Sent: Tuesday, July 27, 1999 11:52:12 AM } } To: Paula Allan-Wojtas; microscopy-at-sparc5.microscopy.com } } Subject: Re: LM- Richardson Real Time Microscope } } } } Hi Paula and everyone, } } I think did do a demonstration on that microscope at the MSC in } Guelph, } Canada. It was very interesting as the manufacturer claims a total } magnification of about 15,000X (this includes TV screen magnification) } from } a light microscope. All the information is proprietary - he would not } tell } me what kinds of modifications he used to make the LM do this kind of } magnification but did say he was willing to visit places and do demos. } At } the meeting, he was affiliated with Leica. I saw some live cells from } asparagus "juice" and could see organelles. I think it makes a bridge } between conventional LM (which generally magnify to 100X) and TEM } (without } all the preparation time). It is interesting enough to look at. I } can't } tell you much more than that as the fact sheet is rather short. All I } can } tell you is that I did see it! } } Lesley Bechtold
At 02:31 PM 7/27/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Somehow, I think that something is wrong with this picture. 100nM resolution and imaging of viruses? With a LM? I am not convinced. OK Microtech, convince me. Maybe they have found out how to make Meiji do wonders.
First of all thanks for all the replies, its been helpful. Second, sorry = about the lack of info here's a bit more of the background. =20
Recently we acquired a Gatan Model 600 Dual Ion Mill, Ultrasonic Disc = Cutter and a Dimple Grinder. I've spent the last couple of months = cleaning the Ion Mill and rebuilding the whisperlocks and guns. I finally = got it working a couple of weeks ago. Although one of the guns on the = room temperature stage is a bit unstable, it appears to sputter in bursts. = I think its probably the Ar supply, either the solenoid valve or the = needle valve. =20
Now what we want to do is start preparing cross sectional TEM samples of = TiN films on Si. Thus we need the wafering saw, to prepare the samples. = Although I don't know if what we need is a precision saw or not. =20
I haven't done anything like this before, so I'm learning on the fly.=20 So any info I can get on the equipment and/or the techniques, ie. books, = papers, etc is a bonus. =20
Thanks again folks
George
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 1793 +61 3 9925 2205 Fax: +61 3 9925 5290 Email: george.theodossiou-at-rmit.edu.au
Home Ph: +61 3 9808 9085
Impossible I Can Do Today, Miracles, Require 24 Hours Notice
Looking at their images the resolution doesn't look any better than what I get with an old Leitz. The contrast is a little better but I bet a good servicing of the old Leitz would help the contrast too.
I am reminded of Rife's microscope. Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger -----Original Message----- } From: Dr. Gary Gaugler {gary-at-gaugler.com} To: MSA listserver {Microscopy-at-Sparc5.Microscopy.Com}
Our sample preparation for TEM involves using silicon 'backing blocks' of dimensions 10mm x 4mm x 1mm. We have a 3 inch diameter (?) silicon ingot which could yield enough blocks to last a lifetime if only we had someone to dice it up for us. Does anyone know of, ideally, a UK based company who could do it?
TIA
Alan Walker
***************************************************************************** Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom Phone: +44-(0)114-2225365 (direct) +44-(0)114-2222000 (switchboard)
I had the opportunity to organize a 2 days demonstration with that = microscope last july on several biological samples (food related one's) = and people including myself where very impress by the resolution, contrast = and the simplicity of the technique that does not require any kind of = staining to improve the image and all this in real time. Emulsion of all kinds, bacteria, yeast, liquids...etc are perfect = candidates for that type of microscopy. Optical sectioning is possible to a certain extent, surface imaging is = also possible.
The software is inexistant and the manipulation of the instrument does = require a bit of training and having a soft fist but it seems more to be = common sense than anything else (very clean area, very good quality slides = and coverslips, clean water, clean is the key word in fact).=20
The instrument itself is composed of a mechanically modified microscope, = improved optics, no oculars, an imaging device of some sort, a good = quality tv screen and a small black box with a few controls similar to pmt = one's.This instrument is indeed a proprietary one for the time being for = patent related reasons, it is not for sale but for rent.
I do believe, like Lesley Bechtold, it does fill a gap between conventiona= l LM and SEM/TEM and that we will hear more and more about that canadian = innovation.
Diane Montpetit microscopist EM lab Agriculture and Agri-Food Canada Food research and development center St-Hyacinthe (Qu=E9bec) Canada J2S 8E3 tel 450 773 1105 fax 450 773 8461 montpetit-at-em.agr.ca
I agree. Have a look at their web site. Do they show anything there that can't be done with phase-constrast LM? Not as far as I can tell. So maybe someone could explain to me the huge advantage?
Rob Palmer CEB/UT
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just a note to let you know that the previously offered filaments are already spoken for. Requests were only seconds apart(!). I'm always still amazed how fast response time is on this thing...
Want to share this guess on the fungus by Gwyneth Beagley at Alma College: "...and the mold is philapusmicroscopus..." Thanks for the smile Gwyneth.
john www.uga.edu/caur/ ******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu
In Needham book, The Practical Use of the Microscope, Second Edition, Chapter IX, pages 123 to 136. It talks about Near Ultraviolet and Ultraviolet Microscope used for photo's with separation of structure lines per inch as high as 250,000 (0.10 Micron) using quartz objective 1.25 NA with a wavelength of 253mu this was back in the 1930 or 40.
Food for thought;
Would it not be possible to use the IR spectrum, quartz objectives and a IR camera to get a high resolution?
Best Regards Joseph Passero jp-at-spacelab.net
Gordon Couger wrote Wed, 28 Jul 1999 02:14:51 -0600 } } Looking at their images the resolution doesn't look any better than } what I get with an old Leitz. The contrast is a little better but I bet a } good servicing of the old Leitz would help the contrast too. } } I am reminded of Rife's microscope. } Gordon } } Gordon Couger gcouger-at-couger.com } 624 Cheyenne } Stillwater, OK 74075-1411 } 405 624-2855 GMT -6:00 www.couger.com/gcouger } } -----Original Message----- } } From: Dr. Gary Gaugler {gary-at-gaugler.com} } To: MSA listserver {Microscopy-at-Sparc5.Microscopy.Com} } Date: Wednesday, July 28, 1999 1:29 AM } Subject: RE: LM- Richardson Real Time Microscope } } } } } } At 02:31 PM 7/27/99 , you wrote: } } } } } } } } } After rummaging a bit on the internet I found the following URL: } } } } } } http://www.bio-microtech.com/products/rtm/ } } } } } } You can check out the PDF file there for more info. } } } } } } We have NO connection to this company or the microscope, so this time I } } } will not include my disclaimer. I just remembered that I had seen it on } } } the web. (Ask me for image acquisition from this scope, though) ;-) } } } } } } Somehow, I think that something is wrong with this picture. 100nM resolution } } and imaging of viruses? With a LM? I am not convinced. OK Microtech, } } convince me. Maybe they have found out how to make Meiji do wonders. } } } }
I just want to thank Bob S, Ron, Joel and Bob P for their help with my "sticky" airlock situation. I tried the simplest solution first and the problem is solved. The more involved replies will be archived and saved for future use. Regards,
Andrew Ochalski, Microscopy Technician, Dept. of Biology, University of Ottawa Room 108, Gendron Bldg. 130 Louis Pasteur, Ottawa, Ontario CANADA K1N 6N5 613-562-5800 x6343 FAX: 613-562 5486
I want to thank all of those who replied to my query, both publicly and = privately. Even Tim Richardson himself telephoned me and explained the = capabilities of this microscope.
I certainly learned a lot about the microscope, and heard from many people = who were very impressed with demonstrations. All of this makes me want to = test it out for myself in the near future!
Thanks again.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
First off, thanks for the list. I'm learning a lot and (I hope) have even been able to contribute a little.
A note to let you know my level of ability. I was trained as a chemist. I've morphed into a metallographer having received training in house at the hands of my self-educated boss. I'm now getting more into the use of SEM and have used LM mainly to evaluate microstructures of metals.
Now to my question. I am doing some consulting work with a pharmaceutical company that is developing a 'new and improved' method to deliver asthma drugs. One of the tasks I've been given is imaging the size of ergotomine tartrate particles which are clogging some of the orifices (orifii?). The VP is hesitant to try using SEM due to the danger of sublimation of the particles. So far light microscopy (low power stereoscopic) has not proven very informative.
I suggested the possibility of an E-SEM but wanted to see what the list would have to say.
Thanks in advance for any help/direction you can supply!
Steve Cavender Metallographer AMPS (Advanced Modular Power Systems, Inc.) 4370 Varsity Drive Ann Arbor, Michigan 48108-2241 734-677-4260 x 209 voice 734-677-0704 fax scavender-at-ampsys.com
To assist faculty and graduate students with technical research tasks involving tissue preparation and imaging of biological specimens using immunocytochemistry, in-situ hybridization, fluorescence microscopy, confocal microscopy and electron microscopy; Conduct short term research pilot-projects to establish feasibility, refine or develop new protocols; Provide ongoing advice and assistance with longer-term projects primarily conducted by members of an individual lab. Help train graduate students in these (or new) techniques; Maintain up to date knowledge and skills by review of literature, consultation, courses, etc. Opportunities to develop and publish new procedures. Contact Dr. John Elam, Neuroscience Program, Department of Biological Science, Florida State University, Tallahassee, FL 32306. {elam-at-neuro.fsu.edu} Neuroscience web page: {http://www.neuro.fsu.edu} , Florida State University is an EEO/AA Employer.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Kimberly A. Riddle Florida State University tel: 850.644.6519 Biological Science Imaging Resource 119 Bio Unit I, 4370 fax: 850.644.0481 Tallahassee, FL 32306 riddle-at-bio.fsu.edu ~~~~~~~~~~~~~~~~~~~~~~~~~~~~
In response to the current discussion on the list server regarding the Richardson Real Time Microscope:
Unfortunately there seems to be a lot of confusion, the RTM microscopes are not related to the low cost RFM microscopes at all (except they were designed by the same design group). The RTMs are expensive extremely high performance imaging systems designed to see inside live cells in real time. They do have final screen magnification of 10,000 to 15,000X with resolution of 180 nanometre and detection limit currently of 50 nanometre. So for instance you can detect, image and track, but not fully resolve, viruses in real time inside cells. The microscopes work as a result of a collection of modifications and improvements to almost every aspect of the microscope. Every part is designed, produced and hand fitted to offer imaging at the limits of the laws of optical physics.
These microscopes have been tested and calibrated using ultrahigh resolution microscope calibration slides with a full range of features from several microns to less than 100 nanometres.
The images from the RTM have been described as looking like super DIC or real time SEM of live samples. We can look at the surface of samples like a sample (like an SEM) or provide optical sections like a (TEM) or a confocal. The system is designed for real time imaging, 60 fields per second, of living cells, bacteria, fungi, or other biological samples in colour without stains or fixation. Sample prep is very fast, typically under a minute. Since no stains or dyes are used to accomplish the high contrast images questions about artifacts due to staining, fixation or other prep methods are virtually eliminated.
The images on the website have been highly compressed to give a reasonable download time so they are not at full resolution. For full resolution images in digital or print form please contact the company. A full resolution image file is very large (over 900K). For reference, a compressed image of unstained live chlamydia in tissue culture is located at "http://www.bio-microtech.com/Gallery/chlamydia.htm".
Bio-Microtech, the company that manufactures the RTM, offers three versions of the RTM technology, the RTM-2: a basic colour brightfield system. The RTM-3: a combined RTM and fluorescent system which allows both images to be superimposed so fluorescent markers can be seen in full context with other cellular components, and the RTM-4: an inverted microscope with similar features to the RTM-2. Each system is supplied as a "turnkey package" including the microscope, S-VHS video system, S-VHS recorder, high resolution monitor and all accessories needed to start immediate work. Each system includes the cost of an intensive three day course.
Now for the RFM.
The RFM is a field microscope designed for superior reliable imaging in rugged use at remote locations. The concept behind the RFM design was to provide a microscope that can be equally useful to a young enthusiastic child of less than 10 or to a seasoned professional field microscopist. The instrument had to be virtually indestructible and yet provide images with the quality and reliability of a good clinical or bench microscope.
The most notable feature of the RFM is the image quality. The RFM uses standard camera tripods as the mount so it is equally at home on the bench or on the side of a rugged river valley. The RFM has a built in illuminator and battery that provide extremely flat illumination. The battery is a standard 9 volt and provides days of continuous use. It can be used at magnifications ranging from 20X to 1000X. It is inexpensive as Caroline Schooley suggests. It is built to last with all parts fully replaceable.
Both the RTM and the RFM where designed by researchers at Bio-Microtech for their own internal use. The RTM was designed to look inside cancer cells and to image bacterial/cellular and viral/cellular interactions. The RFM was designed for family field use. Only later were these systems offered as commercial products.
Hope this clears things up a bit.
Disclaimer:
Tim Richardson directs research at Bio-Microtech Inc, which is the supplier of the RTM and RFM technologies.
Tim Richardson, Director, R&D, Bio-Microtech Inc., Phone: 905-951-7058 Fax: 905-951-7052
There is an immediate opening for an experienced Scanning Electron Microscopy (SEM) Research Technician at Exxon Research and Engineering Company's Corporate Research Laboratory in Clinton, New Jersey. This is a permanent Research Technician position.
As a member of the Advanced Characterization Group at Corporate Research, this position will involve the operation of the state-of-the-art JEOL FESEM, and JEOL Analytical SEM instruments with associated Energy Dispersive and Wavelength Dispersive Spectrometers. The position will involve the creative application of high resolution SEM imaging and elemental characterization of samples related to a wide range of projects at Exxon's Corporate Research Laboratory. The position will also involve general laboratory operations, sample preparation, SEM maintenance and computer analysis of the SEM data (both image analysis and spectral analysis).
Previous SEM experience and knowledge of high vacuum systems and computers (Unix, DOS, and Windows) is required. Successful candidates should have experience in chemistry, physics or material science with a Baccalaureate degree or equivalent experience. Since a number of projects are simultaneously in progress, it is essential for the researcher to be very organized and to pay attention to detail and accuracy in reporting results.
All qualified candidates please send resume to:
Human Resources Recruiting - RTSEM Exxon Research and Engineering Company P.O. Box 998 Annandale, New Jersey 08801-3344
FAX (908)730-3081
All resumes must be received by August 13, 1999.
Exxon is an Equal Opportunity Employer M/F/D/V
(For qualified candidates attending the MSA/MAS conference held August 2-5 in Portland, OR, look for this notice and the Exxon Research & Engineering Co. representative.)
William A. Lamberti Exxon Research & Engineering Company Route 22 East Annandale, NJ 08801 (908)730-2144 (fax = 3314) email "walambe-at-erenj.com"
To the subscribers: I am doing a school project and I need some very general statistics on microscope use--I was referred to this List Server. Any help would be greatly appreciated as soon as possible. I can be contacted at SDHynd-at-aol.com or at 610-745-6170. Thank you.
Joe, Although I have no personal experience with UV microscopes, they would have an advantage in resolution due to the shorter wavelength. This is not without problems as you have mentioned, quartz optics and of course the danger of working with UV and it's potential detrimental affect on samples. For these reasons, cost, and other microscopes (SEM, Confocal) and the small gain in resolution are factors for the lack of success of these scopes. On the other hand IR is going in the wrong direction for any resolution increase. Russ Xerox , -----Original Message----- } From: Joseph Passero [mailto:jp-at-spacelab.net] Sent: Wednesday, July 28, 1999 9:50 AM To: Microscopy-at-sparc5.microscopy.com
In Needham book, The Practical Use of the Microscope, Second Edition, Chapter IX, pages 123 to 136. It talks about Near Ultraviolet and Ultraviolet Microscope used for photo's with separation of structure lines per inch as high as 250,000 (0.10 Micron) using quartz objective 1.25 NA with a wavelength of 253mu this was back in the 1930 or 40.
Food for thought;
Would it not be possible to use the IR spectrum, quartz objectives and a IR camera to get a high resolution?
Best Regards Joseph Passero jp-at-spacelab.net
Gordon Couger wrote Wed, 28 Jul 1999 02:14:51 -0600 } } Looking at their images the resolution doesn't look any better than } what I get with an old Leitz. The contrast is a little better but I bet a } good servicing of the old Leitz would help the contrast too. } } I am reminded of Rife's microscope. } Gordon } } Gordon Couger gcouger-at-couger.com } 624 Cheyenne } Stillwater, OK 74075-1411 } 405 624-2855 GMT -6:00 www.couger.com/gcouger } } -----Original Message----- } } From: Dr. Gary Gaugler {gary-at-gaugler.com} } To: MSA listserver {Microscopy-at-Sparc5.Microscopy.Com} } Date: Wednesday, July 28, 1999 1:29 AM } Subject: RE: LM- Richardson Real Time Microscope } } } } } } At 02:31 PM 7/27/99 , you wrote: } } } } } } } } } After rummaging a bit on the internet I found the following URL: } } } } } } http://www.bio-microtech.com/products/rtm/ } } } } } } You can check out the PDF file there for more info. } } } } } } We have NO connection to this company or the microscope, so this time I } } } will not include my disclaimer. I just remembered that I had seen it on } } } the web. (Ask me for image acquisition from this scope, though) ;-) } } } } } } Somehow, I think that something is wrong with this picture. 100nM resolution } } and imaging of viruses? With a LM? I am not convinced. OK Microtech, } } convince me. Maybe they have found out how to make Meiji do wonders. } } } }
} so what was the simplest solution? } - just for the rest of us with Philips scopes, in case we ever come on } similar troubles... } } Thanks, } } Brendan Foran } SEMATECH } Austin, TX
Greetings,
The problem with the specimen holder that became very difficult to turn in the Philips 201 TEM airlock was solved by a chloroform cleaning of the injector rod (avoiding the rubber o-ring), followed by a light coating of the o-ring with Apiezon M vacuum grease. Simple, but it's nice to have the information from the other postings if this doesn't turn out to be enough. Regards,
Andrew Ochalski, Microscopy Technician, Dept. of Biology, University of Ottawa Room 108, Gendron Bldg. 130 Louis Pasteur, Ottawa, Ontario CANADA K1N 6N5 613-562-5800 x6343 FAX: 613-562 5486
Resolution depends on the wavelength of illumination and the N.A. of the optics.
R = (.61 x wavelength in nm) / N.A.
Thus at short wavelengths (blue and UV) one would have better resolution than with Infrared given the same N.A.
I agree the web site photos don't seem any better than regular DIC.
Date sent: Wed, 28 Jul 1999 09:50:27 -0400 } From: Joseph Passero {jp-at-spacelab.net} Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
In Needham book, The Practical Use of the Microscope, Second Edition, Chapter IX, pages 123 to 136. It talks about Near Ultraviolet and Ultraviolet Microscope used for photo's with separation of structure lines per inch as high as 250,000 (0.10 Micron) using quartz objective 1.25 NA with a wavelength of 253mu this was back in the 1930 or 40.
Food for thought;
Would it not be possible to use the IR spectrum, quartz objectives and a IR camera to get a high resolution?
Best Regards Joseph Passero jp-at-spacelab.net
Gordon Couger wrote Wed, 28 Jul 1999 02:14:51 -0600 } } Looking at their images the resolution doesn't look any better than what } I get with an old Leitz. The contrast is a little better but I bet a } good servicing of the old Leitz would help the contrast too. } } I am reminded of Rife's microscope. } Gordon } } Gordon Couger gcouger-at-couger.com } 624 Cheyenne } Stillwater, OK 74075-1411 } 405 624-2855 GMT -6:00 www.couger.com/gcouger } } -----Original Message----- } } From: Dr. Gary Gaugler {gary-at-gaugler.com} } To: MSA listserver {Microscopy-at-Sparc5.Microscopy.Com} } Date: Wednesday, July 28, 1999 1:29 AM } Subject: RE: LM- Richardson Real Time Microscope } } } } } } At 02:31 PM 7/27/99 , you wrote: } } } } } } } } } After rummaging a bit on the internet I found the following URL: } } } } } } http://www.bio-microtech.com/products/rtm/ } } } } } } You can check out the PDF file there for more info. } } } } } } We have NO connection to this company or the microscope, so this time } } } I will not include my disclaimer. I just remembered that I had seen it } } } on the web. (Ask me for image acquisition from this scope, though) ;-) } } } } } } Somehow, I think that something is wrong with this picture. 100nM } } resolution and imaging of viruses? With a LM? I am not convinced. OK } } Microtech, convince me. Maybe they have found out how to make Meiji do } } wonders. } } } }
Russell N. Spear Sr. Research Specialist Dept. of Plant Pathology Univ. of Wisconsin-Madison
This is a very relevant paper that you might want to look at:
"Transmission Electron Microscopy Characterization of Hard Coatings and Films: Sample Preparation Aspects and Results" G. Radnoczi, A. Barna; Surface Coatings & Technology 80 (1996) 89-95.
This paper does deal with the IV3 Ion Milling System that we sell so I should make my disclaimer here. However, the paper does deal specificall= y with the preparation of TiN on Si so it should be of interest. If you don't have access to the paper, let me know and I'll send one to you.
I also have a bibliography of over 200 papers that deal with sample preparation - I would be happy to e-mail you a copy of the bibliography s= o you could look for any other papers of interest. I would be pleased to send any of them to you (at no charge, of course).
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "George Theodossiou" } --------------------------------------------------------------------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
G'day All,
First of all thanks for all the replies, its been helpful. Second, sorry=
about the lack of info here's a bit more of the background. =
Recently we acquired a Gatan Model 600 Dual Ion Mill, Ultrasonic Disc Cutter and a Dimple Grinder. I've spent the last couple of months cleani= ng the Ion Mill and rebuilding the whisperlocks and guns. I finally got it working a couple of weeks ago. Although one of the guns on the room temperature stage is a bit unstable, it appears to sputter in bursts. I think its probably the Ar supply, either the solenoid valve or the needle=
valve. =
Now what we want to do is start preparing cross sectional TEM samples of TiN films on Si. Thus we need the wafering saw, to prepare the samples. =
Although I don't know if what we need is a precision saw or not. =
I haven't done anything like this before, so I'm learning on the fly. =
So any info I can get on the equipment and/or the techniques, ie. books, papers, etc is a bonus. =
Thanks again folks
George
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 1793 +61 3 9925 2205 Fax: +61 3 9925 5290 Email: george.theodossiou-at-rmit.edu.au {
I found it quite informative to just look at the resolution of a normal microscope. I found the following as a "definition" of resolution (somewhere on the internet, I can probably locate the URL if that is needed):
The first step in this process is to determine the resolving power of the microscope. The ultimate limit on the spatial resolution of any optical system is set by light diffraction; an optical system which performs to this level is termed "diffraction limited." In this case, the spatial resolution is given by: d = 0.61 x lambda / N.A. where d is the smallest resolvable distance, lambda is the wavelength of light being imaged, and N.A. is the numerical aperture of the microscope objective. This is derived by assuming that two point sources can be resolved as being separate when the center of the airy disc from one overlaps the first dark ring in the diffraction pattern of the second (the Rayleigh criterion). It should further be noted that, for microscope systems, the numerical aperture to be used in this formula is the average of the objective's numerical aperture and the condenser's numerical aperture. Thus, if the condenser is significantly underfilling the objective with light, as is sometimes done to improve image contrast, then spatial resolution is sacrificed. Any aberrations in the optical system, or other factors which adversely affect performance, can only degrade the spatial resolution past this point. However, most microscope systems do perform at, or very near, the diffraction limit.
Now, let's plug in the numbers that are given for the Richardson scope on the net:
visible light: about 0.5 microns wavelength N.A. (100x): 1.4
That results in a theoretical resolution of about 210 nm, which is not too far from what they claim as the resolution. However, this resolution should be attainable with any good microscope with a lens N.A. of 1.4.
They then claim a detection limit of { 100 nm. What their definition of "detection limit" is, I don't know. Perhaps some variation of the Rayleigh criterion (see above)?
Regarding wavelength: Going to IR will decrease your resolution (lambda in the formula goes up). Using UV can get you better resolution.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Joseph Passero[SMTP:JP-at-SPACELAB.NET] } Sent: Wednesday, July 28, 1999 7:50:27 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: LM- Richardson Real Time Microscope } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In Needham book, The Practical Use of the Microscope, Second Edition, Chapter IX, pages 123 to 136. It talks about Near Ultraviolet and Ultraviolet Microscope used for photo's with separation of structure lines per inch as high as 250,000 (0.10 Micron) using quartz objective 1.25 NA with a wavelength of 253mu this was back in the 1930 or 40.
Food for thought;
Would it not be possible to use the IR spectrum, quartz objectives and a IR camera to get a high resolution?
Best Regards Joseph Passero jp-at-spacelab.net
Gordon Couger wrote Wed, 28 Jul 1999 02:14:51 -0600 } } Looking at their images the resolution doesn't look any better than } what I get with an old Leitz. The contrast is a little better but I bet a } good servicing of the old Leitz would help the contrast too. } } I am reminded of Rife's microscope. } Gordon } } Gordon Couger gcouger-at-couger.com } 624 Cheyenne } Stillwater, OK 74075-1411 } 405 624-2855 GMT -6:00 www.couger.com/gcouger } } -----Original Message----- } } From: Dr. Gary Gaugler {gary-at-gaugler.com} } To: MSA listserver {Microscopy-at-Sparc5.Microscopy.Com} } Date: Wednesday, July 28, 1999 1:29 AM } Subject: RE: LM- Richardson Real Time Microscope } } } } } } At 02:31 PM 7/27/99 , you wrote: } } } } } } } } } After rummaging a bit on the internet I found the following URL: } } } } } } http://www.bio-microtech.com/products/rtm/ } } } } } } You can check out the PDF file there for more info. } } } } } } We have NO connection to this company or the microscope, so this time I } } } will not include my disclaimer. I just remembered that I had seen it on } } } the web. (Ask me for image acquisition from this scope, though) ;-) } } } } } } Somehow, I think that something is wrong with this picture. 100nM resolution } } and imaging of viruses? With a LM? I am not convinced. OK Microtech, } } convince me. Maybe they have found out how to make Meiji do wonders. } } } }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 any commercial labs that do XRF analysis as a service? } Are there any such labs located in New England? } } N. Carl Miller
When needed, I use Evans East in E. Windsor, NJ for detailed surface analysis of materials (i.e., ESCA, ESCA, TOF - SIMS, Auger, etc.). Call for a complete list of their techniques & pricing (609) 371-4800. Their web site is www.evanseast.com
J. Roy Nelson Material Testing Laboratory Pennington, NJ
Michael, You just about answered your own question.
Resolution is discussed in terms of the minimum detectable separation between two _point_ sources. The sources need have no size (but do in reality). The issue is whether they can be resolved from each other as distinct objects. Much smaller objects may be detected in the microscope - it would just be impossible to tell what their actual size is. I suppose the 50 nm figure comes from knowing the size of a virus particle. An even smaller particle with sufficient contrast with its background should be detectable; however, we would have no idea of its true size from that microscope. We would depend on some other microscopic technique to pull out a size number.
Therefore, I suggest that the 50 nm figure is meaningless and only clouds the issue. Better to simply quote the specification in terms of a resolution figure. Then I can compare the quoted 180 nm for the RTM with the 4 nm guaranteed for our old JEOL 840A and realize that even though the RTM appears to be a good scope, it is far from an SEM in performance.
Warren S.
At 12:31 PM 7/28/1999 -0600, Michael Bode wrote: } This is derived by assuming that two point sources can be } resolved as being separate when the center of the airy disc from one } overlaps the first dark ring in the diffraction pattern of the second } (the Rayleigh criterion).
{snip}
} They then claim a detection limit of { 100 nm. What their definition of } "detection limit" is, I don't know. Perhaps some variation of the } Rayleigh criterion (see above)?
Microscopy/Marketing & Education conducts a considerable amount of market research each year. Please contact me off-line with more specifics.
Thanks,
Barbara Foster Consortium President Microscopy/Marketing & Education - THE primary source for action-catalyzing information in microscopy and related imaging.
125 Paridon Street, Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 Email: mme-at-map.com www.MME-Microscopy.com/marketing ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ MME is America's first national consortium offering full service technical marketing support to the microscopy and imaging industries.
At 01:34 PM 7/28/99 EDT, ThirtyOneT-at-aol.com"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'd appreciate any specific information anyone has about preparing yeast cells for TEM. I have a protocol which removes the cell wall using Glusulase after initial fixation. Unfortunately, I can not find specific concentration information. If you have any experience with this enzyme (or lyticase or Zymolyase, for that matter), I'd appreciate hearing from you.
I'd also appreciate any general advice about thin sectioning yeast cells. We are looking for virus like particles (VLP's) in the cytoplasm.
Thanks,
Chris -- Chris Best, best-at-juniata.edu Mol Biol Juniata College Huntingdon, PA 16652
Indeed, detection and resolution are different. I am inserting a portion of an email I wrote last time this question came up.
{start of insert}
Here is a simple test: go outside at night and look at the stars. Now, individual stars are below the resolution limit of the unaided eye (ex. you won't be able to resolve a binary star system). However, obviously you can see (detect!) them. This is because resolution limits and detection limits are two different things. Think of a single small bright object on a dark background. As the object approaches a true point, the image approaches the point spread function (by definition). There is no reason to expect that as the object drops below some (resolution) limit, the light coming from the object will stop propagating to the detector. As long as it is there is enough light reaching the detector, you can still detect it. The image doesn't get smaller (since the PSF is the limit, and it will get dimmer, but it still can be detected). Resolution simply involves seeing how close _another_ object can get to the first one without their images overlapping by some amount (that amount depending on whether you use the Rayleigh or Sparrow criterea). The perception or detection of a bright object on a dark background is limited by the "brightness" of the object. Now, the perception of a dark object on a bright background is different situation. Consider a dark line on a bright background: what happens when the line narrows and the two line spread functions get close to start to overlap? The minimum will become shallower and shallower. Based on the eyes' ability to discern intensity variations, Pluta (reference below) gave formulas for the detection of these objects: Using typical parameters (488nm, 0.9NA) the smallest dark objects on a bright background that can be observed are 41nm for a disk, and 2.6nm for a line.
Resolution, perception/detection and location are different, but unfortunately all three tend to get lumped together as "resolution". I know you didn't ask, but since it's related, you can also locate an isolated small object to better than the resolution limit (if you know that your system has at worst only symmetric aberrations). Try this: draw a circle and then try to find the center. Remember, this is not a random processes, but a deterministic one. This also extends to edge location. One study (can't find the reference right now) back in 1986 showed that confocal microscopes (of that era) had a 20nm uncertainty in locating edges (the same value as for the SEMs of the day). This means that as long as a the two sides of a line object are resolved, then you can measure the width of the object to much better than the resolution limit. Obviously SEMs can perform the measurements on narrower objects than LM, but I did included the qualification concerning the object being wide enough for the edges to be resolved in the previous sentence. (No flames for saying that LM will replace EM: I stated the limitations twice! (and pointed it out again)) Again, resolution, perception/detection and location are different (related to the imaging system, but still different).
Regards, Matt Atkinson 3M Corporate Research Labs
(Pluta's book Advanced Light Microscopy, vol 1, pg337-348 gives a very good explanation.)
{end of insert}
Michael Bode wrote:
} {- discussion of resolution snipped-} } } They then claim a detection limit of { 100 nm. What their definition of } "detection limit" is, I don't know. Perhaps some variation of the } Rayleigh criterion (see above)? } } Regarding wavelength: Going to IR will decrease your resolution (lambda } in the formula goes up). Using UV can get you better resolution. } } Michael } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215
There is an XRF listserver where you would probably find this information. I think you can subscribe by sending a "SUBSCRIBE" command to LISTSERV-at-LISTSERV.SYR.EDU. If that doesn't work, contact the list administrator (Michael Cheatham {mmcheath-at-MAILBOX.SYR.EDU} ).
Jim McGee
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 any commercial labs that do XRF analysis as a service? } Are there any such labs located in New England? } } N. Carl Miller
Does anybody, possibly vendors, know approximately how many electron microscopes (I am only interested in TEM, STEM and SEM instruments) have been sold in the world (to date) since the first commercial microscope was developed?
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
You can give us a call. While we are not in New England, we do service clients from around the country. Evans East, Charles Evans & Associates and Surface Science Labs are different branches of the same company. Surface Science Labs is located in Mountain View, CA and is the location with the XRF equipment.
Robert J. Plano Staff Analyst, SPM Services Charles Evans & Associates/Surface Science Labs (650)962-8767, ext. 742 http://www.cea.com http://www.surface-science.com
-----Original Message----- } From: "Norman_C_Miller-at-res.raytheon.com"-at-Sparc5.Microscopy.Com [mailto:"Norman_C_Miller-at-res.raytheon.com"-at-Sparc5.Microscopy.Com] Sent: Tuesday, July 27, 1999 3:12 PM To: Microscopy-at-Sparc5.Microscopy.Com
Does anyone know of any commercial labs that do XRF analysis as a service? Are there any such labs located in New England?
My first posting was rather vague--I apologize. Thank you to those who helpd guide me in the right direction. I have received some great help already, but certainly would appreciate any additional info that someone could provide. What I am looking for ( and I now know that this is no small task) is the following (if possible):
Market Size, expenditures, revenues, etc. Market Segments, end users Market growth
I only need overall numbers, I do not wish to obtain any information that is proprietary.
I can be contacted at SDHynd-at-aol.com or at 610-292-9282. Thanks again for your help.
I was reminded of another technique that may of interest which was developed by John McCaffrey and Scott Walck. This is a MicroCleaving technique which has been very effective (and quick) when working with coatings. We do manufacture the MicroCleave Kit so I do have a vested interest in promoting its use. I will send you a data sheet separately. =
For detailed technical discussions on how the technique might be applied = to your TiN/Si samples, I would suggest speaking with Dr. Scott Walck (walck-at-ppg.com). He works magic with this technique and has has been successful with a wide variety of sample sets. I think he also offers workshops on the technique when time permits.
I also have some technical notes on the technique as well as a video whic= h was put together following an MSA tutorial we did a few years ago. Any o= f these are available at no charge if you have an interest.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "George Theodossiou" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
G'day All,
First of all thanks for all the replies, its been helpful. Second, sorry=
about the lack of info here's a bit more of the background. =
Recently we acquired a Gatan Model 600 Dual Ion Mill, Ultrasonic Disc Cutter and a Dimple Grinder. I've spent the last couple of months cleani= ng the Ion Mill and rebuilding the whisperlocks and guns. I finally got it working a couple of weeks ago. Although one of the guns on the room temperature stage is a bit unstable, it appears to sputter in bursts. I think its probably the Ar supply, either the solenoid valve or the needle=
valve. =
Now what we want to do is start preparing cross sectional TEM samples of TiN films on Si. Thus we need the wafering saw, to prepare the samples. =
Although I don't know if what we need is a precision saw or not. =
I haven't done anything like this before, so I'm learning on the fly. =
So any info I can get on the equipment and/or the techniques, ie. books, papers, etc is a bonus. =
Thanks again folks
George
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 1793 +61 3 9925 2205 Fax: +61 3 9925 5290 Email: george.theodossiou-at-rmit.edu.au
Home Ph: +61 3 9808 9085
Impossible I Can Do Today, Miracles, Require 24 Hours Notice {
To etch copper from stainless there is no requirement to expose the sample to the ferric chlorde etch at such elevated temperatures. 20 oC will do the job.
Chris
} Hello Microscopists ! } } Santosh Kurinec asked how to remove copper from stainless steel } without damaging the stainless. Then Chris Jeffree suggested } ferric chloride ... } } Chlorides in general are death to stainless steel - pitting or } stress corrosion cracking can occur, especially if the exposure } occurs at temperatures above about 150 F or if the stainless } steel is being plastically deformed at the same time. } } I would use concentrated nitric acid, which will remove the } copper in an instant. However, some stainless steels are } sensitive to nitric acid (that's the reason there's a Huey } test in ASTM A262) so the exposure should not be for long. } The attack takes place along inclusions on exposed sections } that were transverse to the direction of prior mechanical } working. It's also called, "end grain attack." Stainless } steels with welds in them should also not be exposed to the } concentrated nitric acid for long times. } } This should be done under a hood, of course. } } Nonsusceptible stainless steels are quickly passivated by } the nitric acid, so they shouldn't even etch. } } The same technique works on carbon steel items coated with } copper or with copper parts embedded in them. } } Best regards, } George Langford, Sc.D. (Metallurgy) } amenex-at-amenex.com } http://www.amenex.com/
===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
Does anyboby out there have experience of immunogold labelling of heat shock proteins (HSP) in marine macroalgae? I have found very few references to contemplate and my work to date has been somewhat disappointing.
So far I have produced pretty good TEM images using appropriate immuno preparation protocols, but cannot identify any HSP on my sections. The suppliers of my primary antibodies are confident that they will stick to any HSP present in my tissue. I am studying Enteromorpha, Palmaria and Fucus, the heat shock response has been demonstrated in Enteromorpha by workers here using western blotting methods, and I am following their culture conditions.
Any comments, however basic would be greatly appreciated, as would names of potential contacts in this area of study
Many thanks in advance.
Pete
Peter Bond Plymouth Electron Microscopy Unit University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel/Fax: 01752 233092 email: pbond-at-plymouth.ac.uk
} Date: Thu, 29 Jul 1999 11:48:42 +0200 } From: Fernandes Elisabeth {fernandes-at-univ-paris12.fr} } X-Mailer: Mozilla 4.06 [en] (WinNT; I) } To: sdw-at-biotech.ufl.edu } Subject: Hi } } I am a french student and I would like to know if it already exists some } images and articles about observations unisng TEM of collagen gel which } contains fibroblasts. } I would like to know if it has already been realized some measurement of } porosity of a material using photos realized by TEM. } I hope someone would answer my questions } Thanks... } } } }
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I have done similar SEM work on drug particles accumulating on various plastic & rubber parts of inhalers. I did not have any special problem with sublimation of the drug at the magnifications you will need to see the presences of particles. Standard SEM procedures should work, otherwise give me a buzz off-line.
J. Roy Nelson Material Testing Lab. (609) 730-0575 jrnelson-at-nj1.aae.com
Dear listers,
First off, thanks for the list. I'm learning a lot and (I hope) have even been able to contribute a little.
A note to let you know my level of ability. I was trained as a chemist. I've morphed into a metallographer having received training in house at the hands of my self-educated boss. I'm now getting more into the use of SEM and have used LM mainly to evaluate microstructures of metals.
Now to my question. I am doing some consulting work with a pharmaceutical company that is developing a 'new and improved' method to deliver asthma drugs. One of the tasks I've been given is imaging the size of ergotomine tartrate particles which are clogging some of the orifices (orifii?). The VP is hesitant to try using SEM due to the danger of sublimation of the particles. So far light microscopy (low power stereoscopic) has not proven very informative.
I suggested the possibility of an E-SEM but wanted to see what the list would have to say.
Thanks in advance for any help/direction you can supply!
Steve Cavender Metallographer AMPS (Advanced Modular Power Systems, Inc.) 4370 Varsity Drive Ann Arbor, Michigan 48108-2241 734-677-4260 x 209 voice 734-677-0704 fax scavender-at-ampsys.com
An EM Lab it the Twin Cities (MN) is shutting down and would like to get rid of their equipment....
TEM Microscope: JEOL model JEM-100B. Needs repair. "Vacuum Failure". Possible contaminated column. TEM includes: Edwards Penning 505 Vacuum Monitor, NesLab HX50 Water Chiller, JEE-4B Vacuum Evaporator, and OMAR Critical Point Dryer. Unit is approximately 30 yrs. old (installed in 1971). Buyer would have to de-install.
ULTRA MICROTOMES: 1. Reichert Ultracut E 2. Reichert Supernova 3. LKB Ultratome III 4. LKB 2088 Ultratome V 5. Sorvall JB-4 6. Sorvall and LKB Knife Makers
DIAMOND KNIFES:
There are 3 each DDK Diamond Knifes. Each are 2.5 - 2.9 mm. Two have been used. One has not - box has never been opened.
The MSA web site still lists "tentative speakers" for the '99 Pre Meeting Congress on "Optical Microscopy in the Next Millenium." I would greatly appreciate it if someone could post the final list of speakers, schedule of presentations, and meeting place. Thank you.
Has anyone translated the usual fortran code for Kramers-Kronig dielectric function determination into a Mathematica Notebook or an executable file not requiring a compiler? If so, would they be willing to share? I have found KRAKRO.FOR in Egerton's excellent book and ELSKKT.FOR on the Microscopy FTP site but do not have a compiler, at present. Maybe I am being lazy, but if you don't ask...
Thanks in advance,
Rob Dickerson ********************************************************* Robert M. Dickerson Mailto:dickerson-at-lanl.gov MST-CMS Mailstop K765 Tel: ph:505-667-6337 Los Alamos National Laboratory Fax: 505-665-2992 Los Alamos, NM 87545 TA-03 Bldg.1698 Rm.C-136 *********************************************************
Can anyone help out this person? Please reply directly to
Robert Byrnes {rbyrnes-at-abac.com}
} The CPD that I am trying to bring back up to usuage is a BOMAR } SPC-900/EX. The last listed address for the company is The Bomar Co., } P.O. Box 225, Tacoma, Wa 98401. So far I have not been able to find } out if this company still exists. Any information would be a help. } Thanks.
To remove Cu from stainless steel without the use of corrosive acids, make up a mixture of 1 part conc. ammonia, 1 part water, and 1part hydrogen peroxide (30%). Under a fume hood, wash the part with this solution. It will foam up and turn blue. Rinse and repeat until the Cu Is removed. Do not re-use the swab or cleaning cloth. The presenceof the blue compound will cause the wash solution to decompose immediately.
Sam Purdy
} ---------- } From: Chris Jeffree } Sent: Thursday, July 29, 1999 6:05 AM } To: amenex-at-amenex.com; microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Copper etch (removal of copper from stainless steel) } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have been tasked with investigating feasibility of calcium ratioing for some of our researchers. Not knowing much about it, I wanted to see if anyone has any experience with systems doing this.
What manufacturers systems are recommended/not recommended? (Please reply privately if you feel you will offend manufacturers publicly).
Specifically, why do you recommend/not recommend that configuration?
What should I look for/look out for when asessing calcium ratioing systems? (Features, functionality, capabilities, etc.)
Food Structure and Functionality -=20 An international symposium taking food structure studies into the 21st = century held in conjunction with the=20 SCANNING 2000 Meeting, May 9-12, 2000 at the Sheraton 4 Points Motel, San Antonio, Texas, USA
Symposium Activities: -Short course - Food Microscopy - Identifying Food Ingredients and = Contaminants given by James Charbonneau (National Food Processors = Association, USA) and Mark Auty (TEAGASC, Moorepark, Ireland) , assisted = by John Shane (McCrone Institute, USA) Carol Kennedy (Feed Microscopist, = Agro Pacific Industries, Canada), and Kathy Groves (Leatherhead Research = Association, UK). is scheduled for Tuesday 9 May 2000, 8:30 am - 5:00 = pm=20
- 3 full exciting days of oral and poster scientific sessions including = commodities, techniques, instrumentation, applications and Agriculture are = planned, covering the full spectrum of food structure from production to = processing.=20 The preliminary program for the 3 days of sessions (Wednesday, May 10th to = Friday, May 12th inclusive) is in the planning stages.=20 Session topics include, but are not limited to:=20 Water structuring - covering gelling and thickening Foams and emulsions - covering topics in interface science Agricultural Applications - covering food production and quality Dairy applications Meat and Fish Applications Cereals applications New methods and instrumentation for food structure/texture measurement
- Mixers and other social functions=20
-A meeting of our interest group, the Food Structure and Functionality = Forum =20 The Forum's mandate is:=20 To promote global collaboration between Food and Agriculture professionals = in Structure and Functionality disciplines by facilitating interaction and = providing a forum for exchange of knowledge, expertise and research = findings.
--Short courses offered during the SCANNING meeting on topics including = microscopy, image analysis, forensic microscopy =20 -Manufacturer's exhibition
More details will follow soon and will be continually updated- keep = watching the Foods under the Microscope website: http://www.cyberus.ca/~scimat/fsf.htm =20
SCANNING website:
http://www.scanning-fams.org=20
Or, you can contact one of the Forum Core members: Paula Allan-Wojtas Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5 Tel: (902) 679-5566 FAX: (902) 679-2311 email: allanwojtasp-at-em.agr.ca =20
Mark Auty Dairy Products Research Centre TEAGASC Moorepark, Fermoy Co. Cork Ireland Tel: 011 - 353 - 25-42447 FAX: 011 - 353 - 25 - 42340 email: mauty-at-moorepark.teagasc.ie
James E. Charbonneau National Food Processors Association Food Chemistry and Packaging Department 1401 New York Ave., NW Washington, D.C. 20005, U.S.A. Tel: (202) 639-5972 FAX: (202) 639-5991 email: jcharbonneau-at-nfpa-food.org
N.C. Ganguli National Academy of Agricultural Sciences Indian Dairy Association M-76B, Malviya Nagar New Delhi 110017 India Tel: 011 - 877 - 0650 or 011 - 618 - 5355 FAX: 011 - 332 - 0613 or 011 - 696 - 8433
A while back I posted a message regarding an apparent enhancement of CL from quartz due to electron beam bombardment. That is, if the quartz emitted CL, it could be enhanced by allowing the beam to raster the area for some period of time .. and not all luminescing qtz grains will exhibit this phenomenon. Somewhat interesting, but more a pain-in-the-A**, because the mere act of zooming in for a focus would put a bright rectangle in the middle of an interesting luminescent quartz grain. We have since made additional observations and can pass them on ...
First, many of you suggested the enhancement could be the result of contamination. This is not the case. The fact that all quartz will not behave this way should have been an initial clue, but we have since subjected these quartz grains to repolishing and recoating, and they will still show us the original enhanced rectangles. We don't know to what depth the quartz has been apparently "bothered", but we definitely removed some material by apply the final 0.25u diamond phase of normal polishing.
We have since added a capability for filtering the CL thru red, green and blue filters ... and we have also added a PMT which extends out wavelength sensitivity into the UV. We have a couple of quartz projects in the works which I can't discuss generally, but with regards to the modest varieties of quartz we have looked at this year we have noticed a preponderance of quartz which luminesces red and those which luminesce in the UV. We have noticed the above mentioned induced enhancement occurs only with those grains which emit red, and if the grain emits a component of UV/blue and red, then the induced enhancement shows only in the red part of the spectrum.
For those who are also studying CL in quartz and who might also be aware of this phenomenon, we invite your comments in hopes of realizing an explanation.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I just receive with my correspondance one ad about Fujifilm FDL 5000 system. Just for curiosity, there is someone that has experimented this system and the phosphor plates? How does it compares with the standard films and a negative scanner? I am pretty sure that a FDL 5000 is quite expensive.
Regards,
Kazuo
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
I recollect from a year or two ago comments on the quality and durability of different coloured dyes on recordable CD's for archival storage of images. Has anyone any up to date information on whether one colour is worse than the others.
Thanks in advance
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hort.cri.nz
Dear Hildy Crowley, Im referring to your posting from the 9th of July. During the last time I was not only bringing up the collegues against me with my statements ;-(, I also was doing some quite good immunos on thin sections. I had the luck to made an extraordinary good LR-White embedding with high-resulution fine structure and good contrast on whole heart tissue. First, I made an immunolabeling with an membrane associated antigen. It worked well, the signal was weak but significant. I was quite happy. Then I tried to visualize SNAP-25, which is thought to play an important role in membrane fusion, even in endothelium...but this approach failed.
Conventional LM immunos on parafine and vibratome sections, developed with HRP/DAB failed, too. I used Antibodies from Tranduction Labs. Up until now, I never had a real signal to SNAP-25. So my question to you is, if you had positive light-microscopic control results with SNAP-25 and what kind of antibody you are using. I think our work is very similar in this special case.
Good luck and the best wishes Yours, Michael "the Junior" Reiner
I am also interested in the final list and schedule.
-----Original Message----- } From: Dan Luchtel [mailto:dluchtel-at-u.washington.edu] Sent: Thursday, July 29, 1999 11:28 AM To: microscopy-at-Sparc5.Microscopy.Com
The MSA web site still lists "tentative speakers" for the '99 Pre Meeting Congress on "Optical Microscopy in the Next Millenium." I would greatly appreciate it if someone could post the final list of speakers, schedule of presentations, and meeting place. Thank you.
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Many of us have accepted high contrast negatives from metal or ceramic specimens as normal and unavoidable. Alas, there may be a better way! I will begin with a disclaimer for the products mentioned below. I have no vested interest in them-but they have made my work easier. A widely used developer for Kodak SO-163 film is Kodak's D-19 diluted 1:1 with water. It is occassionally used full strength for "push" processing. When exposures are made of complex structures in metal foil specimens, its high contrast tends to hide some fine detail, especially in the darker areas of the negative. Use of low contrast print paper, manual dodging, or computer scanning are used to "salvage" as much info as possible. To alleviate the problem, automatically exposed, identical test negatives were made. When developed in Kodak D-76 diluted 1:1 with water, negatives had a noticably lower contrast and could be printed on F-2 grade paper with minimal dodging for good results. The cost of D-76 is similar to D-19, but the "tank life" as diluted is only a day or two. More tests were done from the stockpile of identically exposed negatives with a two bath "split" developer named Diafine. It's made by Acufine, Inc., 5441 North Kedzie Avenue, Chicago, Il.,60625. In effect, this developer "chemically dodges" the contrast of the negative. It requires placing the film in tank A for 3 minutes, then in tank B for three minutes. Times and temperatures are not critical. The resulting negatives printed very well on F-2 paper with no dodging. This developer has a tank life at least as long as D-19, costs about two times as much, and increases the film speed about 0.5 F/stop. The darkroom time and print paper saved should make it a bargain in the long run while producing good results. When printed with a Log E computerized enlarger these negatives make excellent prints. For a dramatic test, a TEM negative exposed normally, was cut in half. One portion was developed in conventional D-19, the remainderin Diafine. The Diafine processed film was far easier to print properly and retained more detail of the specimen's structure. This approach may result in too little contrast for biological specimens-but a couple of test exposures will reveal that. Sometimes two exposures of an area of particular interest may be taken. One is developed in D-19, the other in Diafine. The negative showing the desired characteristic is then used. This technique may help those who don't have computer scanners. (I learned of Diafine from Nestor Zaluzec). Bernard Kestel, Materials Science Div., Argonne Nat'l. Lab, 9700 South Cass Ave., Argonne, Il.,60439. E-mail {kestel-at-anl.gov} Phone: (630) 252-4945
by cats.ucsc.edu (8.8.6/8.8.4.cats-athena) with SMTP id PAA04865 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 29 Jul 1999 15:09:48 -0700 (PDT) X-Sender: jmkrupp-at-cats-po-1.ucsc.edu Message-Id: {v01550102b3c68084b326-at-[128.114.25.148]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Hi:
A colleague has requested an SEM picture of a CD ala the discussion a few weeks ago here. If you have a picture you would be willing to share, you may contact him to work out the details. If you prefer, I will be at MSA and you could track me down there and I will take it to him.
Here is the contact info.:
Kenneth Coale, Acting Director Moss Landing Marine Laboratories P O Box 450 Moss Landing, California 95039 USA voice (831) 755-8655 fax (831) 753-2826 Shipping address: 895 Blanco Circle Salinas, California 93901 USA home page http://color.mlml.calstate.edu/www/
Thanks.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have a colleague who is having problems getting some tissue to stain. The material was fixed in 1% GLA/PB 7.4, washed in H2O, dehydrated through ethanols and embedded in Leica Historesin (GMA). The sections are 5u thick and are of CNS of a teleost. They are proving to be very resistant to staining. He's tried the usual thick section stains (methylene blue, toluidine blue O) and cresyl violet, with and without heat. Nothing seems to be working well at all--any suggestions? Grace
Has any-one used p-phenylenediamine dihydrochloride with pyrocatechol (PPD-PC) on LR White sections at the LM level, and them taken them to the EM level? Any tips, hints or ideas on this would be greatly appreciated. Susie Nilsson
Balazs Labs in the SF Bay area is hiring an SEM/EDX operator. Please contact Jeff Sanders at 408 745 0600 ext 19, or jsanders-at-balazs.com (I have no connection with Balazs, I'm just posting the want ad for them!) Bill Neill
I have sold a camera from the company "PIXERA". They have a model 120es which give you very descent pictures in fluorescence.
This camera has a resolution of 1260 x 960 lines, and is less expensive than the Spot2. Try to visit their homepage: http://www.pixera.com If you are interested, let me know. I sell and adapt this camera to maby different microscopes. With small CCD, the optical adaptation to a microscope, is sometimes as important than the camera itself.
Best regards,
Emile Meylan
emeylan-at-csi.com
----- Original Message ----- } From: Blancaflor, Elison {eblancaflor-at-noble.org} To: 'Microscopy-at-MSA.Microscopy.com' {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, July 27, 1999 7:28 AM
G'day all,=20
Thanks for all the information, the response has been great. =20 I'm sure that I'll have futher questions, but at this point I'm going to = take a couple of days to digest the information and talk to the boss about = it. =20
You'll probably hear from me again sometime in the future. Again thank = you. =20
George
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 1793 +61 3 9925 2205 Fax: +61 3 9925 5290 Email: george.theodossiou-at-rmit.edu.au
Home Ph: +61 3 9808 9085
Impossible I Can Do Today, Miracles, Require 24 Hours Notice
"Nanostructured Materials" session at Materials 2000 Royal Agricultural College, Cirencester, Gloucs, UK 12-14 April 2000
Just a message to let you know about a session on "Nanostructured Materials" being organised by Alfred Cerezo (Oxford), Wendy Vine (DERA) and Rik Brydson (Leeds) at the Institute of Materials Congress "Materials 2000".
Current topics include: Keynote Lecture (Prof Dick Siegal) Structural Nanomaterials Functional Nanomaterials Electronic Nanomaterials Biomimetic Self-assembled Nanostructures Modelling of Nanomaterials Nanomechanics in Composites and Laminates Production Technologies Microscopy/ spectroscopy of Nanomaterials
This will include both invited speakers and contributed oral and poster presentations.
Brochures for the congress are available from Melanie Boyce at Institute of Materials (Email: Melanie_Boyce-at-materials.org.uk) meeting. Provisionally the Nanostructured Materials one day session is arranged for the Friday 14 April and day registration is available.
We are actively seeking contributions (both ORAL and POSTER) !!!
The brochure for the meeting includes a registration form and instructions for submitting abstracts. Please note that the abstract deadline is September 30th 1999. We are encouraging all contributors to the session to submit a written version of their paper for publication in Materials Science and Technology. All the papers will be reviewed in the normal way and will appear in a special double issue devoted to the sessions within the "Structure of Materials" theme.
As happened at the last IOM Congress, the EPSRC have agreed generous funding to all students funded under the Materials Programme to cover their attendance at the meeting. This funding is already agreed, and is separate from the usual funding of one conference per studentship which is normally allowed under EPSRC rules. I therefore hope you will encourage as many of your students as possible to attend the Congress, and also to submit poster papers to our session. _____________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
To all who have responded to the list of equipment that I posted yesterday, please bear with me as I am in the process of collecting the requested specifications. I should have more detailed information soon and will respond ASAP.
Thanks for your patience,
-- Ford M. Royer, MT(ASCP) Analytical Instruments, Ltd (Refurbished Histology, Cytology, & General Lab Equipment) 9921 13th Ave. N. Minneapolis, MN 55441-5004 800-565-1895 phone 612-929-1895 fax web site: http://www.aibltd.com
We are looking to purchase a used TEM in fully operating condition. Microdiffraction capabilities and a double tilt holder are required. EDS capabilities would be helpful. Please respond directly to:
I would like to gather information on "cold light", for use with a light microscope. I am intending to work with frozen samples and a source of heat like the lamp or the light beam itself can make it more complicated. What is generally understood as cold light? Just IR filters or a separated lampcase with optical fibber connections to light the microscope? And, is this kind of illumination correct for all kinds of microscopes/optical systems? At the moment, I am only considering bright field and simple polarisation, as a contrast-enhancement for ice crystals. } I would like to hear the opinion of somebody working with it, because vendors are discouraging me from trying to adapt this cold illumination, and I don't know if the cause is that it is not a good idea or that they don't have it readily available for sale.
Frank Scheltens and I experimented with the imaging plates when I was at Wright Patterson Air Force Base. We ran an exposure test over the range of exposures quoted in their literature and two sheets of film by multiple exposures by moving the SAD aperture around. We used the current meter on the viewing screen to determine the electron exposures. We also did the same thing with regular film for a comparison. The net result was that this stuff is great. The literature says that the density is linear over 4 to 5 decades of exposure and we confirmed this. We also shot some CBED patterns and an image of a sample that had high Z and low Z material in the sample. I could get all the information that was in the sample that I needed. If I remember correctly, the ultimate resolution is about 25 um which is getting pretty close to film.
The interesting thing was that it took us some time to figure out which of the two files corresponded to the higher dose plate and which was the lower dose plate because of the way the files were displayed. In other words, the two plates looked the same even though the exposures were shifted by about 2 orders of magnitudes. We had no problems with the film, the highest exposure on the low dosage film plate (SO-163) just barely registered any density. This also agreed with there published literature which shows the range for SO-163.
I believe that we set up the microscope by adjusting the intensity so that the meter just registered the minimum current. Our minimum exposure corresponded to the minimum exposure time of the microscope (.12sec). Our maximum time was 5 min. Somewhere in there we had to increase the intensity of the beam so that we wouldn't have to wait forever. Our exposures were calculated to stay in the range that is quoted in the literature but at the same time span the whole range. We had information on all the "squares" of exposure (six on each plate) and the maximum did not saturate the plate.
You do have to retrain your "metered eye" especially for diffraction patterns. You have to go to much smaller times than you are normally accustomed to. One problem with the plates though is that you do not get the information written by the microscope onto film because light erases the plates. Bumper. The files are also huge and are 16 bit files (I think they are 14bit deep).
You used to be able to try a few plates out and have them developed by sending them packed in dry ice. I don't know if that is available anymore.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Carlos Kazuo Inoki To: MSA Listserve -----------------------------------------------------------------------.
Hi,
I just receive with my correspondance one ad about Fujifilm FDL 5000 system. Just for curiosity, there is someone that has experimented this system and the phosphor plates? How does it compares with the standard films and a negative scanner? I am pretty sure that a FDL 5000 is quite expensive.
Regards,
Kazuo
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
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It seems to me that this option is currently the best solution for TEM digital imaging. Of course, you don't get your image immediately as with a camera but from what I can see the resolution, sensitivity, linearity and field of view are all excellent and better than any digital camera system. An added advantage for those with more than one TEM, for a relatively small additional cost for extra plates, you can convert as many TEMs as you have all in one hit.
Regards,
-- Larry Stoter JEOL (UK) Ltd Silver Court, Watchmead, Welwyn Garden City, AL7 1LT, United Kingdom tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
Dear Antonio, if you illuminate anything with light and it has a finite absorption coefficient it is bound to warm up, it's just the laws of physics! I would suggest that you use a light filter that allows you to pass frequencies that the ice does not absorb very well. Firstly you need an absorption spectrum of ice. Secondly look for frequency bands where transmission is best and try to find filters that fit within these bands, use overlapping filters if necessary. Getting rid of Infra Red will be a big help in the first place. I would not believe any salesman that gives you a complicated (=confusing) picture of how their product seemingly works. Even if you maintained a vacuum shield or any other fancy trick, the light has got to get through. It would be wiser if you concentrated on keeping the specimen stage cold, and maybe using cold dry nitrogen/dry air to prevent water condensing onto your specimen.
Regards, Jonathan ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
Please contact evex analytical 609-252-9192 jeol and hitachi TEM available
On Fri, 30 Jul 1999, Valdemar Furdanowicz wrote:
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