I know this has been asked before but I cannot easily lay my hand on the information.
What is the current knowledge about the longevity of ink-jet prints (black and white and colour) from medium to low end ink-jets (eg Epson 640 or Phote EX, HP 710). How much difference does the paper make to the expected life of the print.
Thanks
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
Ian Since these products are recent, their permanence in storage has not yet been tested. My Epson 800/1520 prints stored in the dark seem to be OK *so far*. However, I have noticed fading of displayed Epson 800 and 1520 prints on Epson Glossy Photo paper over a period of about 15 months. These prints are mounted on a boared in an internal corridor, exposed to fluorescent lighting only, and daylight exposure would no doubt further accelerate fading. The assumption should be that these madia are not of archival quality, and it would probably be unwise to sell the prints as artworks, for example.
You might like to know that Marrutt (of darkroom rotary door fame) market Lyson long-life inks and media for Epson ink-jet printers Information about this can be found on the following url
http://www.marrutt.com/
Photo buffs may relish the fact that black inks can now be purchased in selenium and sepia tones, in addition to neutral black. Media Available include 120 and 170 gsm Matt Paper, 180 gsm Gloss Paper Backlight film and fine art papers in different surfaces and grades.
I have no experience of these products, and would be interested to hear any users' observations on colour and print quality, etc. when using these products.
Yours sincerely Chris Jeffree
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 know this has been asked before but I cannot easily lay my hand } on the information. } } What is the current knowledge about the longevity of ink-jet prints } (black and white and colour) from medium to low end ink-jets (eg } Epson 640 or Phote EX, HP 710). How much difference does the } paper make to the expected life of the print. } } Thanks } } 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 }
===================================================================== 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 =====================================================================
} Date: Fri, 27 Aug 1999 08:39:03 -0400 } From: Larry Allard {l2a-at-ornl.gov} } Subject: Re: Plasma cleaner } To: "Erasmus, Willem (WJ)" {willem.erasmus-at-sasol.com} } Cc: microscopy-at-Sparc5.Microscopy.Com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } WJ: } } How are your catalyst sample supported? If you disperse the powder over } "clean" holey carbon grids (which contain no residual plastic film from the } production process), by simply dipping the grid into the catalyst powder } and shaking off the excess, you should find that contamination is not a } problem. Typically, powder specimens dangling over holes in carbon films } do not offer enough surface over which to diffuse molecules that cause } contamination buildup under the beam. Of course, the other source of } contamination could be the microscope itself, but that's another story... } } We have found that a plasma cleaner for the TEM specimen/specimen holder } works very well in the instances that we have contamination problems with } fine probe analysis using our Hitachi HF-2000. Our cleaner happens to be a } Fischione, and a couple of minutes treatment generally suffices to cure the } problem. If you suspect that the grids are the source of the } contamination, you might want to clean a grid in the plasma cleaner prior } to deposition of the catalyst specimen. I would be surprised if these } efforts did not produce satisfactory results for you. } } Good luck... } } Larry
Please, be careful not to use the plasma cleaner on holey carbon grids as, to my experience, often the carbon film itself is removed.
Albert Dr. Albert Romano-Rodriguez Professor Titular Dept. of Electronics Faculty of Physics University of Barcelona c/ Marti i Franques, 1 E-08028 BARCELONA Spain tel: +34-93-402 90 69 (+34-93-402 11 47) FAX: +34-93-402 11 48 e-mail: romano-at-el.ub.es
You are quite right. I forgot to mention this...but it turns out that with our Fischione we can plasma-clean holey carbon films for 1-2 minutes and still preserve the film. I think it is gone after about 5 minutes under standard cleaning conditions however. It may be the case that with just an inert gas, say nitrogen, there would be some cleaning benefit also, with perhaps not as much effect on the carbon film, since oxygen is eliminated as the reactive component in the plasma.
Larry
} } Please, be careful not to use the plasma cleaner on holey carbon } grids as, to my experience, often the carbon film itself is removed. } } Albert } Dr. Albert Romano-Rodriguez } Professor Titular } Dept. of Electronics } Faculty of Physics } University of Barcelona } c/ Marti i Franques, 1 } E-08028 BARCELONA } Spain } tel: +34-93-402 90 69 (+34-93-402 11 47) } FAX: +34-93-402 11 48 } e-mail: romano-at-el.ub.es
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
} } What is the current knowledge about the longevity of ink-jet prints } (black and white and colour) from medium to low end ink-jets (eg } Epson 640 or Phote EX, HP 710). How much difference does the } paper make to the expected life of the print.
I can't speak for these specific printers, but archival inks and papers are available for the Epson printers ... e.g., see:
www.dygraphics.com ... or search the wwweb for "lysonic inks"
I have Jim Mancuso's (AMT) older system that used a Kodak 1K 8 bit camera. It has worked fine but is developing hick-ups due to age (purcased 5 years ago) and incompatability with new computer's hardware/software. I reviewed his new system at the MSA 99 meetings and was impressed with it's design (and adaptability to some of my current hardware) and ease of use. His responce to problems has usually been good and he now has additional people working for him so I imagin he won't have to respond to all the calls himself now. Jim has always been good at "working with you " to solve hardware/software, merging your equipment with his, and to come up with a system you can afford. Your lucky, I just wish some one would come up with some money to help me get a 2K system... heck, even one of the new 1K systems!
In the past I too have had problems with getting info from Gatan sales people. I did get a good responce from this last meeting on a specimen holder though.
To be fair, I also looked at Soft Imaging System's and it's design was also good and the poster showing a comparison of their cameras showed very detailed images. I don't know how their price compares with the others though.
As I mention with AMT's system, ask about how well anyones system now will upgrade their older systems so you don/t have to buy a whole system every couple of years. Good Luck.
Roberts, Walck, Grant and Zaluzec conducted some experiments on plasma cleaning crushed zirconolate on holey carbon grids (MRS Spring Meeting 1997) and found the technique to be quite useful. The sample was processe= d in pure argon for a short time which removed most of the contamination. = As the plasma cleaner had 2 independent gas inlets, they were able to perfor= m a final short cleaning with pure oxygen which improved the sample without=
damaging the film. They were also able to reduce the power on the system=
to better control the cleaning process. I suspect that you were using a fixed power machine with a single gas inlet and an argon/oxygen mixture. =
Using that configuration it would be easy to "over process" the sample an= d remove the holey carbon grid. If you still have a need to clean samples = on a holey carbon grid, I would suggest running with the argon and then switching the gas line to run with pure oxygen for a short time. If you cannot reduce the power on your unit, you will still want to be very careful not to over process the sample.
You can find more details on the cleaning of holey carbon grids on our we= b site at www.southbaytech.com/pc150.html. On that page you will find a lin= k to the relevant paper under "Materials Research Society Spring-97 Meeting= ". I also have several other papers on plasma cleaning, plasma trimming and=
plasma etching available (some of those are on the web site). I would be=
happy to send you copies of any or all of them if you have an interest.
DISCLAIMER: The above mentioned work was done with a South Bay Technology= , Inc. Plasma Cleaner. Consequently I have a vested interest in promoting=
Ann No. Nile red is a lipid stain, therefore will stain the wax (though it may have trouble penetrating it to any depth). Wax-embedded tissues will in any case have non-polymerized lipids extracted during processing, so the answer is that Nile red is really applicable only to fresh tissues, deembedded tissues where e.g the cuticle remains after processing - it stains cuticle v. well - or tissues embedded in very polar media (PEG?). It is great for confocal microscopy on live cells and tissues
Chris
} I assume that you are talking about wax embedded tissue. Would Nile red work on resin embedded tissue? } } Ann Fook } } Ann Fook Yang } EM Unit, } Eastern Cereal and Oilseed Research Centre, } Rm 2091, K.W. Neatby Bldg., } Central Farm, } Ottawa, Ontario, Canada K1A 0C6 } } Phone: 613-759-1638 } Fax; 613-759-1701 } } } } } "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} 08/29 2:17 PM } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } I strongly recommend that you use the fluorescent lipid dye Nile } red instead of Sudan Black. Nile red partitions into lipids rendering } them intensely fluorescent. You may use either FITC or RITC filter } sets, as you prefer. What makes Nile red preferable to the Sudan } dyes is that there is virtually no fluorescence from the dye unless it } is in a lipid environment. Therefore the specimen may be mounted } in an aqueous solution containing a low concentration of the dye, } and de-staining the non-lipid parts of the specimen is completely } unnecessary. Furthermore, because the specimen is immersed in } a stock of fluorochrome, there is minimal photobleaching. } } Chris Jeffree } } Date sent: Fri, 27 Aug 1999 09:44:47 -1000 } To: Microscopy-at-sparc5.microscopy.com } } From: "Melany H. Chapin" {mchapin-at-ntbg.org} } Subject: Sudan Black B } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I am looking for a formula and procedure to check plant material for the } } presence of lipids using Sudan Black B. The formula I am using now stains } } everything black. I am specifically checking palm flowers and fruits. Thank } } you in advance. Please reply to my email: } } } } mchapin-at-ntbg.org } } } } ________________________________________________________________________ } } } } Melany H. Chapin Herbarium (PTBG) } } Curator & Plant Records Manager ph: 808-332-7324 ext. 133 } } National Tropical Botanical Garden (NTBG) fax: 808-332-9765 } } P.O. Box 340 www.ntbg.org } } End of Papalina Road email: mchapin-at-ntbg.org } } Lawai, Kauai, Hawaii 96765 } } USA } } ___________________________________________________________________________ } } } } } } } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Dr Chris Jeffree } University of Edinburgh } Biological Sciences EM Facility } Daniel Rutherford Building } King's Buildings EDINBURGH EH9 3JH } Tel: +44 (0) 131 650 5345 } FAX: +44 (0) 131 650 6563 } } Inveresk Cottage, 26 Carberry Road, } Inveresk, Musselburgh, Midlothian EH21 8PR, UK } Tel. +44 (0) 131 665 6062 / Mobile 0410 585 401 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } }
===================================================================== 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 =====================================================================
This is an informative web site I stumbled across last year with comments about ink jet printers, inks and links to digital imaging info. I just checked and it still exists. Updated March 1999.
} } } I know this has been asked before but I cannot easily lay my hand } on the information. } } What is the current knowledge about the longevity of ink-jet prints } (black and white and colour) from medium to low end ink-jets (eg } Epson 640 or Phote EX, HP 710). How much difference does the } paper make to the expected life of the print. } } Thanks } } 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 }
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
If your interested in cleaning Carbon support films, my experiences have shown that a pure Argon plasmsa works very well.
I have used this on holey carbon and lacey carbon films with a variety of materials (crushed powders, catalysts, etc...)
Processing time varies with the degree of contamination and power/geometry of the unit. I prefer to use low power (~ 10 W) and longer processing times (5-10 minutes) this way you don't run the risk of destroying the support film on a processing time which is too fast. Remember you can clean a sample many times in small increments. The experimental data indicates that successive cleaning in small increments is effective.
If a film is particuliarly dirty I might use a few minutes of pure Oxygen, but as mentioned by the previous postings, the Oxygen is very aggressive to the carbon support.
Having a unit that allows multiple gas sources is advantageous in this respect.
Nestor Your Friendly Neighborhood SysOp
Disclaimer: ANL holds the patent on Plasma cleaning for TEM/AEM/SEM specimens and receives royalities from commerical sales of this technology.
I have a client who wants to look at biofilms existing on middle ear mucosa biopsies. The principle constituents of the biofilm are Pseudomonas, Strep. pneumon., and Haem. influ. I have referred him to the U Fl. Tips and Tricks page, but I am sure I have seen a thread on biofilms; I just didn't save them. Does anybody remember when this thread appeared so I can go check the archives or do you have a proven protocol that works? I'm not sure if he wants them with or without capsules. There is a group at Montana St. who do this using a cryo stage apparently because they so many bacteria. TIA for your help. -- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
Hi everyone, Anyone (else) out there have a cryo-stage on the SEM that does service for "outsiders"? I have a project I may not be able to handle due to internal problems with our cryo system {EMSCOPE SP2000A} ie: vacuum leak somewhere in LN2 lines to the stage as well as sputter head lack of coating even with plasma evident in the prep. system. We modified our JEOL 5800LV SEM to fit the system and had it operational ... years ago ( {gulp} ). I am also swamped...ah love the university beginning-of-semester-you-must-do-all-my-microscopy-in-one-month time. =) Thanks!!!
Tracey
Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
I work with a research and development group that prototypes semiconductor processes. Presently, there is a need in my company for a critical point dryer that can accomodate 8" wafers, an exceptionally large chamber size from what I have seen so far. If anyone has any knowledge as to the expandability of critical point dryers, or a source from which such a dryer could be purchased, please let me know.
Thermal microscopy is getting to be big business. You can now do thermal studies with Environmental SEM (ESEM, from Phillips), AFM (ex: Digital Instruments, ThermoMicroscopes, and newcomer JEOL), and light microscopy (see Mettler for a variety of stages and inquire of Leica to see if they still make the elegant VacuTherm, a chamber which sits on an inverted metallograph and permits all sorts of thermal and atmospheric controls).
See also: American Lab, July, 1999 for my review of this sort of instrumentation from the recent PITTCON meeting. If you can't find acopy in the UK, just drop me an email and we'll send you a Xerox.
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 01:13 PM 9/1/99 -0500, Chengge JIAO wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I received many helpful recipes for preparing bacteria for SEM analysis. These are all very informative and useful. Some can be performed by me using what I have at hand.
However, I had subsequently asked for sources/suppliers of prepared specimen stubs for my SEM and wound up with only one potential source.
Am I correct in concluding that no one will prepare SEM specimens of bacteria on a fee basis? Thus, I have to do it myself? What about non-bacterial specimens? Mites, lice, etc.? I have to do these as well?
If no one will provide this service, this helps me justify the investment in the necessary coating, CPD and other equipment necessary to make these specimen preparations.
Seems rather odd that no one does this. I guess that this is reality in this regard.
University of Queensland Postdoctoral Research Fellow Department of Mining, Minerals and Materials Engineering and the Centre for Microscopy and Microanalysis
Applications are invited for the above position. The Department of Mining, Minerals and Materials Engineering (MMM) and the Centre for Microscopy and Microanalysis (CMM) are looking to expand our efforts in the examination of phase transformations in ceramic materials. In this regard, we are specifically interested in the olivine to spinel transformation that is thought to be responsible for the production of deep earthquakes.
To aid us in this project, we are seeking an experienced Electron Microscopist (transmission electron microscopy) with a postgraduate research degree (preferably a PhD) in either materials science, physics, chemistry or earth sciences. The candidate should also have some knowledge of basic crystallography.
This is a full-time position for up to two years and is externally funded via an ARC grant. The successful candidate will be jointly attached to the MMM and the CMM. Salary will be in the range of $41,199.15 - $45,910.62 per annum. We are looking to fill this position as soon as possible. For further information, including position description, selection criteria please contact Dr. John Drennan, Centre for Microscopy and Microanalysis, Telephone: 61-7-3365 6353, Fax 61-7-3365 4422. E-mail: j.drennan-at-mailbox.uq.edu.au.
Closing date for applications 24th September, 1999.
-- ************************************************************ Duncan Waddell (BSc) Senior Scientific Officer Centre for Microscopy and Microanalysis The University of Queensland. St. Lucia. Qld. 4072 Telephone: +61-7-3365-4216 Facsimile: +61-7-3365-4422 WWW: http://www.uq.edu.au/nanoworld/nanohome.html ************************************************************ Any opinion expressed is that of the writer, and not necessarily that of CMM or of the University. ************************************************************
I would like to thank everyone for the helpful, competent and sometimes friendly replies to my posting. I now have a few different options to start working on the preparation of my samples, and some new techniques to exploit too.
Thanks a lot
Massimo
Dr. Massimo Catalano member of the Boards of Directors of Italian Society for Electron Microscopy CNR-IME Campus Universitario Via Arnesano 73100 Lecce - ITALY tel: + 39 0832 322362 * fax: + 39 0832 325299 * email: massimo.catalano-at-ime.le.cnr.it http://www.ime.le.cnr.it http://www.ime.le.cnr.it/sime/sime.htm
* Please note that, effective June 1998, a zero has to be dialed right after the country code (39) before the city code (832).
Tracey Pepper wrote: ======================================================= Hi everyone, Anyone (else) out there have a cryo-stage on the SEM that does service for "outsiders"? I have a project I may not be able to handle due to internal problems with our cryo system {EMSCOPE SP2000A} ie: vacuum leak somewhere in LN2 lines to the stage as well as sputter head lack of coating even with plasma evident in the prep. system. We modified our JEOL 5800LV SEM to fit the system and had it operational ... years ago ( {gulp} ). I am also swamped...ah love the university beginning-of-semester-you-must-do-all-my- microscopy-in-one-month time. =) Thanks!!! ======================================================= Our firm was either the first or among the first independent laboratories, at least in the USA to offer a cryo-SEM service some very many years ago. 100% of our laboratory services business is for "outsiders". We now operate with an Oxford system interfaced to a JEOL Model 840 SEM. We have had experience over the years with just about all the various kinds of samples one might want to look at this way. We would be happy to quote you on your requirements. More information about our services can be found on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI Structure Probe, Inc. FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Prices of SEMs can vary from free (for an old "fixer-up") to over a half million -for top of the line equipment with all the accessory goodies/analytical attachments available.
I'd like to add Nanonics Imaging, Ltd. to your list of providers of Scanning Thermal Microscopy (AFM). While Nanonics is primarily known for NSOM applications, it provides capabilities in Thermal, ElectroChemical, and Chemical Scanning Probe Microscopy.
Sincerely, Dave Eglinton Eglinton Instruments Specializing in Advanced Technologies representing . . . Nanonics Imaging, Ltd
PO Box 2686 Kennebunkport, ME 04046 T: 800-673-2430; F: 207-967-8741; E: eglinton-at-int-usa.net
Barbara Foster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Chennge, } } Thermal microscopy is getting to be big business. You can now do thermal } studies with Environmental SEM (ESEM, from Phillips), AFM (ex: Digital } Instruments, ThermoMicroscopes, and newcomer JEOL), and light microscopy } (see Mettler for a variety of stages and inquire of Leica to see if they } still make the elegant VacuTherm, a chamber which sits on an inverted } metallograph and permits all sorts of thermal and atmospheric controls). } } See also: American Lab, July, 1999 for my review of this sort of } instrumentation from the recent PITTCON meeting. If you can't find acopy } in the UK, just drop me an email and we'll send you a Xerox. } } 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 01:13 PM 9/1/99 -0500, Chengge JIAO wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } Hi, } } } } I am looking for the name and email address of companies who can make or } } supply the High Temp. Micro-scope and Liquid Metal Density Tester. } } } } Does anybody know above items, please send email to: } } } } c.jiao-at-bham.ac.uk } } } } Thanks. } } } } } } } } } }
We need to get information about CL Detectors for a grant proposal. The application has to be submitted within two days, and has just arrived to us.
Does anyone have any information about CL detector types and cost ? Any help would be much appreciated as it will help us to meet the deadline. Information directly from vendors would be very welcome.
Please respond directly to me off list.
Thanks,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
Hello, Has anyone seen circles ~1.4 um diameter , after immunofluroescence in lipofectamine transfection of HeLa cells with the corressponding DNA? What are these structures and what might it colocalize with?
We need to clean our tantalum final (fixed) aperture in a Hitachi 2460 variable pressure SEM. Since they are quite expensive ($600-800) does anyone know of a safe and effective procedure to clean such disc apertures? We are considering heating in a vac evaporator using a tungsten boat. But this procedure is tricky.
Thanks,
John B.
#################################################################### 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 ####################################################################
A colleague is getting ready to service two Porter Bloom MT2-B ultramicrotomes, and I would appreciate if someone would provide the name, phone or whatever contact information you may have of independent service engineers for these microtomes.
Thank you.
John B.
#################################################################### 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 ####################################################################
Are these in the cytoplasm? Or all over the place (in/on cells and the coverslip)? I haven't used any of the "new and improved" lipid-based transfection systems, but we used to get lipid droplets in/around HeLa...the cytoplasm sometimes was packed with them!
Have you tried electroporating, FU-Gene, adenovirus, etc. instead of lipid? All are cleaner (in terms of background) in our hands and the efficiency is higher. BUT, again, I haven't tried lipofection lately.
No financial interest in any of the above methods (I should be so lucky!).
Tamara Howard CSHL
On Thu, 2 Sep 1999, MICHAEL DELANNOY wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hello, } Has anyone seen circles ~1.4 um diameter , after immunofluroescence in } lipofectamine transfection of HeLa cells with the corressponding DNA? What } are these structures and what might it colocalize with? } } Thanks, } } Mike D } } }
Hi. I am running a freeze fracture experiment. I have been tried to get a double replica device for a while. We have a Cressington freeze fracture CFE-50 but we don't have a Cressington double replica kit. I was told that Cressington doesn't supply double replica kit anymore. I talked to Balzers distributor in NH this morning. It seemed they had no idea what I was talking about. Does anybody there know where I can get a double replica device which fits Cressington freeze fracture CFE-50? Any ideas and suggestions would be appreciated. Thanks.
Shanling Shi Unilever Research US 45 River Road Edgewater, NJ 07020 Email: Shanling.Shi-at-unilever.com
I also am looking for a someone with cryo-optical and possibly cryo-SEM capabilities, preferably in the Chicago area. This will be for a potential project looking at several polymer and wax-like materials. thanks, Brad Huggins, EM Lab BPAmoco Naperville, IL
} ---------- } From: Tracey M. Pepper[SMTP:tpepper-at-iastate.edu] } Sent: Wednesday, September 01, 1999 3:54 PM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: cryo-sem } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi everyone, } Anyone (else) out there have a cryo-stage on the SEM that does } service for } "outsiders"? I have a project I may not be able to handle due to internal } problems with our cryo system {EMSCOPE SP2000A} ie: vacuum leak somewhere } in LN2 lines to the stage as well as sputter head lack of coating even } with } plasma evident in the prep. system. We modified our JEOL 5800LV SEM to } fit } the system and had it operational ... years ago ( {gulp} ). I am also } swamped...ah love the university } beginning-of-semester-you-must-do-all-my-microscopy-in-one-month time. =) } Thanks!!! } } Tracey } } } } Tracey Pepper } Supervisor } Bessey Microscopy Facility } Iowa State University } ph: 515-294-3872 } fax: 515.294.1337 } }
} We need to clean our tantalum final (fixed) aperture in a Hitachi 2460 } variable pressure SEM. Since they are quite expensive ($600-800) does } anyone know of a safe and effective procedure to clean such disc apertures? } We are considering heating in a vac evaporator using a tungsten boat. But } this procedure is tricky.
Dear John, The Handbook of Chemistry and Physics says, "Tantalum is almost immune to chemical attack at temperatures below 150 [deg] C ... At high temperatures, tantalum becomes much more reactive." Heating in a vac- uum, where there are no chemicals to react, should be OK, but I'd be sure to let the aperture cool completely before airing the chamber, and I'd make sure all the parts in the evaporator were clean to begin with. I'd plasma- clean to get rid of organics, and use NH4OH for W-residue without heating the aperture. Yours, Bill
Hello everyone - Another lurker pokes his head up out of the foxhole. We are beginning or Fall Semester Biological TEM course and one of the women signed up for the course just discovered that she will be a mother in the fullness of time. It is my opinion - and only that, really- that a biological EM labe with four or five fixation/dehydration/infiltration/embedment cycles proceeding at all times, some in cacodylate buffer, some in s-collidine, or whatever, plus exposure to resin components, etc., is no place for a small human being that is developing very rapidly, even if the packaging is sturdy and intelligent. We have had two other cases in the remembered past where this was a question and both women made an informed decision to stay in the class, against the advice of both the lab director and me. Both delivered healthy, full term babies with no problems (one of the "babies" is at least 13 years old now).
What do the rest of you think is the best course of action? Have there been decent studies of EM lab associated teratogens? Since I have never been a parent myself, this is one of those health hazard questions that I have simply decided to be very conservative about - but I have no hard and fast data that back me up.
Thanks very much for any help - data OR informed opinion that the group comes up with.
William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (602)-965-3210 Fax - (602)-965-6899
} Fall Semester Biological TEM course and one of the women signed up for the } course just discovered that she will be a mother in the fullness of time. } It is my opinion - and only that, really- that a biological EM labe with } four or five fixation/dehydration/infiltration/embedment cycles proceeding } at all times, some in cacodylate buffer, some in s-collidine, or whatever, } plus exposure to resin components, etc., is no place for a small human } being that is developing very rapidly, even if the packaging is sturdy and } intelligent.
If you assembled the MSDS's on all the chemicals used for bio- logical specimen preparation, I'm sure you would be horrified at their potential for harm. Given that ethanol, perhaps the least harmful of the organic solvents, can cause problems--fetal alcohol syndrome if there is enough exposure--I'd be extremely cautious with the more harmful chemicals. Also, many of the chemicals have the specific pur- pose of denaturing (fixing) biological material. The only saving grace--if one can call it that--is that some of these chemicals are so reactive that they will never get as far as the fetus. In this case, they would just damage the mother directly; damage to the fetus would be indirect. Have the woman in question read all the MSDS's before deciding. Remember also that the radiation exposure limits for preg- nant women are at least an order of magnitude lower than for the rest of the population, so the same might be expected for chemical exposure.
} We have had two other cases in the remembered past where this } was a question and both women made an informed decision to stay in the } class, against the advice of both the lab director and me. Both delivered } healthy, full term babies with no problems (one of the "babies" is at least } 13 years old now). } A very small N, and the control experiment was not done. We do not know how much subtle damage was inadvertantly done. Since Pb in very small amounts will lower the IQ a small, but measurable amount, it can be assumed that other environmental insults can also have small effects. Would the expectant mother want to take a year off her child's life expectancy? Increase the chances of cancer by 1%? Such potential problems are not possible to assess from the fact that one child has survived to adolescence.
} What do the rest of you think is the best course of action? Have there been } decent studies of EM lab associated teratogens? Since I have never been a } parent myself, this is one of those health hazard questions that I have } simply decided to be very conservative about - but I have no hard and fast } data that back me up.
I'd (again) get the MSDS's, look up the referrences listed on them, show all this to the woman, and let her decide. But, make sure she makes a fully informed decision. Yours, Bill Tivol
we are exploring the use of our new on-axis CCD camera on our Philips CM200 200 kV FEGTEM.
High resolution images are excellent but we have difficulty with low magnification modes and with capturing diffraction patterns due to the small field of view and the dynamic range in the pattern.
We are interested in conventional and convergent beam patterns.
If anyone out there has solved these problems please respond.
Thanks,
***************************************************** Mel Dickson, Deputy Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
A colleague of mine is exploring the purchase of a used SEM with (or without) EDS. They would prefer a more compact design but would consider others (such as a JEOL 840, for instance). If anyone out there has a system that they've been thinking of getting rid of, and it is in working condition, please contact me directly at the email address shown below. Think of what a good excuse this would be to upgrade your current system!
Cheers, JSV *************************** John Vetrano Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
I have problems with sintering B4C and stabilized zirconia together to obtain dense ceramic materials. A lower processing temperature (e.g., 1500=9AC) is favorable and the reaction between B4C and zirconia should be avoided. I sintered the compact pellet of B4C/ZrO2 mixed powders either in air at 1450=9AC (oxidation was insignificant) or in a hot isostatic press a= t 1250=9AC. But the products were very porous. Any suggestions are welcome an= d appreciated.
Weiliang Gong
Center for Radioactive Waste Management Advanced Materials Laboratory University of New Mexico 1001 University Blvd. SE Albuquerque, NM 87106 (505) 272-7142 (O) (505) 2727304 (FAX) } wgong-at-unm.edu (email address)
William, This issue has been addressed in our laboratories, where there is risk of infection as well as hazardous chemical exposure. As long as all standard precautions are observed and PPE is available for each task, there should be no exposure. All hazards must be addressed in your written procedures and safety criteria must be enforced. Make sure that your fume hood is certified and used correctly, and that gloves are chemical resistant and fit properly. Your institution should have a dept. of environmental and occupational safety and health. They can be a good source for information and policies. Bob Santoianni Emory University Hospital Atlanta, GA
John: For many years we have had Bill McGee of MTS service our Porter Bloom MT2-B ultramicrotomes and can recommend him for the prompt and always satisfactory service. He can be reached by phone at (315) 451-1404. His address is Microtome Service Co. 7568 Florian Way Liverpool, NY 13088 ######################################################## Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
Gloves, and a fume hood and some common sense are all that is needed
At 02:29 PM 9/2/1999 -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 "
Bill: I would like to add to your comments. A not uncommon mistake when cleaning moly or tantalum apertures is overheating, because we all know that these metals have very high melting points and so white-heat seem alright. Unfortunately high heat leads to large crystal formation and the aperture becomes useless after a couple of heating cycles. I used to clean moly and tantalum apertures on a moly (or tungsten) boat. This too should be previously heated for cleaning. Apertures may be stored in a strong solvent and rinsed in ethanol and than water prior to heat cleaning. I suppose the plasma ashing may do that better. Heat cleaning should be for a couple of minutes, but only at a cherry red heat. Leave a couple of minutes to cool before venting the system.
I don't know that aperture, but the appear rather pricey - has John tried to get it trough another supplier? Most suppliers can get just about any aperture made to order, at usually lower prices than EM manufacturer's apertures. Disclaimer: ProSciTech supplies apertures. 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 Friday, September 03, 1999 6:29 AM, William Tivol [SMTP:tivol-at-wadsworth.org] wrote: } } John J. Bozzola wrote: } } } We need to clean our tantalum final (fixed) aperture in a Hitachi 2460 } } variable pressure SEM. Since they are quite expensive ($600-800) does } } anyone know of a safe and effective procedure to clean such disc apertures? } } We are considering heating in a vac evaporator using a tungsten boat. But } } this procedure is tricky. } } Dear John, } The Handbook of Chemistry and Physics says, "Tantalum is almost } immune to chemical attack at temperatures below 150 [deg] C ... At high } temperatures, tantalum becomes much more reactive." Heating in a vac- } uum, where there are no chemicals to react, should be OK, but I'd be sure } to let the aperture cool completely before airing the chamber, and I'd make } sure all the parts in the evaporator were clean to begin with. I'd plasma- } clean to get rid of organics, and use NH4OH for W-residue without } heating the aperture. } Yours, } Bill }
Recording diffraction patterns using 12 bit dynamic range or more should preserve sufficient information in the pattern, though it may be invisible to the human eye. Hence, you'll need to perform some image analysis in order to filter out the bell-shaped background. See e.g.
S. Zaefferer and R. A. Schwarzer Z. Metallk. 85 (1994), p585.
A customized scintillator (or fluorescent screen) for the camera may be used in order to reduce the strong transmitted beam intensity. Alternatively, a customized beam-stop may be used.
The magnification problem may be solved by recording overlapping images from different parts of the diffraction pattern and then stitch them together using image correlation matching. This procedure can be fully automated, though it will require some efforts in software development (CM remote control and image processing). I believe the CM200 is capable of imaging diffraction patterns at very low camera lengths. It may therefore be a better option to reduce the distortion in these patterns by applying a suitable deconvolution procedure.
Cheers, Paul
=================== Paul Baggethun Alcoa Technical Center Alcoa Center, PA 15069 USA =================== + 724 - 337-1760 (tel) + 724 - 337-2044 (fax) ===================
NOTE: The opinions expressed here represent the opinions of the author and do not necessarily represent the opinions of those who hold other opinions.
} ---------- } From: Melvyn Dickson[SMTP:M.Dickson-at-unsw.edu.au] } Sent: Thursday, September 02, 1999 6:47 PM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Diffraction patterns with on-axis CCD camera } } Dear All, } } we are exploring the use of our new on-axis CCD camera on our Philips } CM200 } 200 kV FEGTEM. } } High resolution images are excellent but we have difficulty with low } magnification modes and with capturing diffraction patterns due to the } small field of view and the dynamic range in the pattern. } } We are interested in conventional and convergent beam patterns. } } If anyone out there has solved these problems please respond. } } Thanks, } } } ***************************************************** } Mel Dickson, } Deputy Director. } Electron Microscope Unit, } University of New South Wales. } Sydney NSW 2052 Australia } } Phone (+612) 9385-6383 } Fax (+612) 9385-6400 } Website {http://srv.emunit.unsw.edu.au} } ***************************************************** }
Hi there, does anyone know of a supplier of Teledyne Hastings=20 thermocouple gauges in the MidWest? I ma looking for a replacement=20 for a DV-23 Vacuum Gauge Tube from a Gatan Ion Miller. I can buy=20 this part from Gatan, but I was wondering if I could avoid paying any=20 markup by going a more direct route. Any info would be appreciated
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 been trying in vain for the past two weeks to download the most recent version of UTHSCSA's ImageTool program. The two links I have been trying from a variety of approaches are: http://ddsdx.uthscsa.edu and ftp://maxrad6.uthscsa.edu. Are these still valid, are there more current links or is the software no longer available ? Thanks for any information on this matter. 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 am interested in etching thin sputtered palladium films approx. 500-1000 angstroms thick. would you please assist me in finding the appropriate chemical to etch with. i would like to have an etch time less than one minute.
As you know Ladd manufactures and cleans thousands of moly and tantalum apertures for numerous applications besides electron microscopes. Some of those applications, such as for satllites, x-ray or medical equipment, require ultra-clean apertures where even the appearance of transient debris is a problem.
In the hope that some of our experience might be of benefit let me tell you what we do:
1. We have a proprietary chemical pre-cleaning process after the aperture is manufactured.
2. We then heat the aperture in a tungsten boat in our evaporator. We feel it's critical in this process to: a) Heat only till cherry red for approximately 5 minutes. b) Turn the current up slowly and back down slowly. c) Be sure the aperture returns to its normal color before venting to air.
3. We will also store and ship in acid cleaned containers.
Hope this helps in some way.
Disclaimer: Ladd does manufacture and sell evaporators and apertures in a variety of materials for electron microscopes.
John Arnott -- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
I am looking for an instruction manual for an Reichert-Jung model ''Histoslide 2000'' slinding Microtome. Any suggestions? Thanks. I look forward to your responses.
JOSE ULLAN SERRANO --------------------------- Departamento de ANATOMIA Universidad Panamericana c/ Donatello, 59 Col. Insurgentes-Mixcoac 03920 MEXICO, D.F.
Hello, I have a couple echants you might want to try. 1) Aqua Regia in Glycerol, used cold. I am not sure of the ratios. 2) Another fast etch is hot Nitric Acid. I am sure there are other etches but many employ some fairly noxious chemicals. Here is an example of a slow etch which can be speeded up with the addition of a 3% solution of Sodium Iodide. Potassium Cyanide 20% in water and Amonium Persulphate 20% in water.
Good luck and be careful. This information was gleaned from some dusty old notes.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone: 512/282-5507 FAX 512/280-0702
QUALITY ELECTRON MICROSCOPE SERVICE -----Original Message----- } From: Corcelius, Brian {BCorcelius-at-krautkramer.com} To: 'microscopy-at-msa.microscopy.com' {microscopy-at-Sparc5.Microscopy.Com}
The Imaging Technology Group at the Beckman Institute, University of Illinois at Urbana-Champaign is seeking a research specialist in light microscopy who will be responsible for operation, maintenance and training for the light microscopes in the ITG microscopy suite.
Responsibilities will include:
=B7 Operating/maintaining light, fluorescence, confocal and stereology microscopes. =B7 Application development: Work in conjunction with users to appl= y new LM techniques to their research. =B7 Supervise and train others in the use of the light microscopy instruments. =B7 Develop novel applications to take advantage of the unique capabilities of the instruments.
More information on facilities and instrumentation can be found at http://www.itg.uiuc.edu/
This position requires a Bachelor's degree with experience in all aspects of light microscopy (specimen preparation, imaging, data analysis). Desirable skills would include experience with data analysis packages for light microscopy.
This is a 12-month, 100% time regular appointment with standard university benefits. Salary will be commensurate with education and experience. At this time, the position is available for a period of three years. The position is available immediately. In order to ensure full consideration, applications must be received by September 21, 1998. Please send letter of application and resume to:
Tricia Ware Imaging Technology Group Beckman Institute for Advanced Science and Technology 405 N. Mathews Urbana, IL 61801 (217)244-0170 e-mail: pware-at-uiuc.edu
The University of Illinois is an Affirmative Action/Equal Opportunity Employer. Women and minorities are encouraged to apply.
Stephen Rogers University of California at Davis Section of Molecular and Cellular Biology Briggs Hall, Room 149 Davis, CA 95616 tel: 530.752.2273 email: steverog-at-life.uiuc.edu
Dear John, According to my sources in Hitachi Service Canada, this should be a Molybdenum aperture and should be cleaned by the standard high-vacuum, cherry-red heat in a Mo boat. The Mo boat should be heated empty first to clean it. This method should also work on a tantalum aperture. At 02:22 PM 9/2/99 -0500, you wrote:
} } We need to clean our tantalum final (fixed) aperture in a Hitachi 2460 } variable pressure SEM. Since they are quite expensive ($600-800) does } anyone know of a safe and effective procedure to clean such disc apertures? } We are considering heating in a vac evaporator using a tungsten boat. But } this procedure is tricky. } } Thanks, } } John B. } 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
Vetrano, John S wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi All; } } A colleague of mine is exploring the purchase of a used SEM with (or without) } EDS. They would prefer a more compact design but would consider others (such as } a JEOL 840, for instance). If anyone out there has a system that they've been } thinking of getting rid of, and it is in working condition, please contact me } directly at the email address shown below. Think of what a good excuse this } would be to upgrade your current system! } } Cheers, JSV } *************************** } John Vetrano } Pacific Northwest National Laboratory } MSIN P8-16 } P.O. Box 999 } Richland, WA 99352 } Phone: (509)372-0724 Fax: (509)376-6308 } Email: mailto:john.vetrano-at-pnl.gov
We sell SEM's, TEM's and related Instruments. Available: AMRAY 1200 $4,900.00 1200B 9,000.00 1700, LaB6/W, turbo pumped $19,000.00 Vacuum Evaporators, Sputter coaters, Crit. Point Dryers and other equipment. Regards, Markus F. Meyenhofer Microscopy Labs
John F. Mansfield wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi there, does anyone know of a supplier of Teledyne Hastings } thermocouple gauges in the MidWest? I ma looking for a replacement } for a DV-23 Vacuum Gauge Tube from a Gatan Ion Miller. I can buy } this part from Gatan, but I was wondering if I could avoid paying any } markup by going a more direct route. Any info would be appreciated } } 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° 16' 48" Long. 83° 43' 48"
John,
You can buy DV-23 vac. gauge tube from Kurt J. Lesker Company (vacuum products): (800)245-1656 - lesker.com
Disclaimer: I have no business affiliation with the KJL Company other than being satisfied customer.
I think UTHSCA ImageTool was taken over by Scion Image(???) At any rate, Scion has an easy to use image analysis program that can be downloaded for free from their site:
http://scioncorp.com
Sometimes I have LUT problems, but not when I'm doing image analysis. (Before I got Photoshop, I used to use it to draw pictures ;-) )
-------------------------------------
On Fri, 3 Sep 1999, Andrew Ochalski wrote:
} } Dear Microscopists, } } I have been trying in vain for the past two } weeks to download the most recent version of } UTHSCSA's ImageTool program. The two links } I have been trying from a variety of approaches } are: } http://ddsdx.uthscsa.edu } and } ftp://maxrad6.uthscsa.edu. } Are these still valid, are there more current links } or is the software no longer available ? Thanks ..
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Shanling Shi: I know that double replica devices were available years ago. I still don't know why, so if you require one for your project, let me know. This is how I understand the FE technique and the replica device.
During the FE process a very cold blade removes a series of thin shavings from the sample frozen specimen, to expose a surface of an internal cross section. This results not in a smooth cut (on a microscopic scale) but follows lines of natural weakness, often over and under organelles. "Etching" in Freeze Etching, refers to a short time period, when the much colder knife (minus 170, versus the specimen at minus 100 degrees C) is "parked" above the specimen and some ice sublimes onto the knife, thus increasing relief. Subsequently the samples is coated with Pt/C at an angle and C to make the replica. After cleaning the replica is viewed. These replicas give a surprisingly detailed and somewhat three-dimensional impression of the specimen, which is not prepared using conventional fixation. Now - Shadows from the Pt coating are not electron dense and appear to us negative, meaning "hills and valleys" are reversed. For this reason FE images were often printed as negatives. If you were to prepare one negative and one positive print you would have the impression of the two matching sides, like from a double replica device.
The only reason for a double replica device I can think off, is that the fracture would be much coarser than the one exposed by conventional FE "shaving" method. With the event of high resolution SEM. which has partially replaced the need for the elegant, but difficult FE (TEM) method, there is basically no need for the double replica device. I saw no other posted reply on this topic, but would love to learn from users of the double replica device if there are any special merits today.
Disclaimer: ProSciTech handles some FE technique supplies, but not the apparatus. 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 Friday, September 03, 1999 3:54 AM, Shanling Shi [SMTP:Shanling.Shi-at-unilever.com] wrote: } } } Hi. I am running a freeze fracture experiment. I have been tried to get a } double replica device for a while. We have a Cressington freeze fracture CFE- } 50 } but we don't have a Cressington double replica kit. I was told that } Cressington } doesn't supply double replica kit anymore. I talked to Balzers distributor in } NH this morning. It seemed they had no idea what I was talking about. Does } anybody there know where I can get a double replica device which fits } Cressington freeze fracture CFE-50? Any ideas and suggestions would be } appreciated. Thanks. } } Shanling Shi } Unilever Research US } 45 River Road } Edgewater, NJ 07020 } Email: Shanling.Shi-at-unilever.com }
For what it's worth: I heard Gladys Fiedler speak at a meeting of the AAAS in 1991 about the topic of teratogens and motherhood, and interestingly, she said that the only known case at that time of working with teratogenic materials actually affecting a fetus was that of a man who had worked with heavy metals and his child was born with some kind of defect. At the time Dr. Fiedler was speaking, the government was making some rule that women of child-bearing age couldn't even work in certain jobs for fear that they might affect their offspring, and the point she was making - probably because, as I understand it, this was (is?) her area of study - was that teratogenic affects are perhaps even more likely to affect the sperm which is constantly being made than to affect the child in the uterus, unless of course the woman ingests the teratogen. Just a thought. Gayle Brosnan-Watters
Gayle Brosnan-Watters, Ph.D. Assistant Professor Psychology Department Vanguard University of Southern California 55 Fair Drive Costa Mesa, CA 92626 Phone 714-556-3610 Ext. 454 Fax 714-966-6316 GBrosnanWatters-at-vanguard.edu
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Dear Microscopists, } } I have been trying in vain for the past two } weeks to download the most recent version of } UTHSCSA's ImageTool program. The two links } I have been trying from a variety of approaches } are: } http://ddsdx.uthscsa.edu } and } ftp://maxrad6.uthscsa.edu. I had the same difficulty. I finally got it at http://macorb.uthscsa.edu/dig/itdesc.html.
I did not "trace" the freeze-fracture discussion. In my point of view, people used "double replicas" technique when for some reason they want to see complementary surfaces of the sample. Double replicas device was available many years ago from BALZERS. Simplest construction is two copper/brass small plates with joint. Plates may rotate by hard spring 180 degree in the manner of the book's cover when you open the book. In the start position, the plates are closed (like closed book) with sample between the plates (say pages in the book) and fixed in such position by some kind of "stopper". The "book" than quickly frozen and mounted in the vacuum evaporator. Vacuum evaporator equipped with some simple device to release "stopper" in time. After all necessary manipulation, "stopper" is released then plates are "opened" by force of the hard spring and breaks the frozen sample. With good luck, the sample broke in the middle than both plates exposure the part of sample with complementary surfaces. The surfaces are shadowed and so as usual. I think, such device is so easy to make in machine shop. Described "book"-like device was developed many years ago in the Institute of Biophysics, Pushchino, Russia. I don't know who was author of this device, but it was used in the Lab. Prof. Borovyagin. I newer used such device in my experiments but was frequent "witness" how people used that. If my description of the "device" is not clear (it is better to draw it), please, E.mail me and I will share with you all I know in this matter. If people from ListServer interested, I may prepare some drawing to illustrate the principle and sent it privately upon request.
Best regards,
} Date: Sun, 05 Sep 1999 22:42:05 +1000 } From: jim {jim-at-proscitech.com.au} } Subject: RE: Freeze fracture Need Help } To: 'Shanling Shi' {Shanling.Shi-at-unilever.com} , } "Microscopy-at-MSA.Microscopy.Com" {Microscopy-at-sparc5.microscopy.com} } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } Organization: ProSciTech } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Sergey Ryazantsev Department of Biological Chemistry UCLA School of Medicine Box 951737 Los Angeles, CA 90095-1737 Phone: (310)825-1144 (Lab) FAX (departmental): (310) 206-5272 mailto: sryazant-at-ucla.edu mailto:sryazant-at-ucla.edu http://www.bol.ucla.edu/~sryazant E. mail: sryazant-at-ucla.edu http://www.ben2.ucla.edu/~sryazant
I have a problem obtaining diffraction patterns on our CCD camera. We have a Philips CM200 TEM fitted with a Gatan Image Filter. Due to an extra magnification factor introduced by the GIF the image on the TEM viewing screen is much smaller than the image seen on the computer screen. Although bothersome, this does not present too much of a problem in ordinary imaging.
However, when obtaining a diffraction pattern I find that one cannot shield the CCD camera from the centre spot (transmitted beam) with the beam-stop because the image of the beam-stop covers the diffraction pattern. The only thing visible on the screen is the beam-stop.
At the moment I am recording diffraction patterns on film, but is there a way to use the CCD camera for this purpose ?
Dear members of the list, I think I remember that some food researchers are on the list. They could help me or anyone of you with a better biochemical background or somebody who knows other sources which could help me on this question:
Who can give me some general information on caramels and caramelization? What influences the fluidity/viscosity of caramel? Are there ways to induce caramelization at lower temperatures than 65=B0C?
Thanks in advance and best wishes for you all, Michael Reiner
I am looking for the recipe for Dalton's Osmium Solution. I have a couple of references, but the details of the recipe are different. Does anyone have the citation of the original reference? Thank you very much!
Hello, This may seem a strange solution but why not use a smaller beam stop? All the beam stop is is a wire of Aluminum. You could make another beam stop of a lesser diameter. Try pulling some Aluminum wire. You can get a very small diameter, provided the alloy has enough ductility. I would suggest that it is not good idea to blast your CCD camera with the strong diffracted beam.
Good luck.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone: 512/282-5507 FAX 512/280-0702
QUALITY ELECTRON MICROSCOPE REPAIR
-----Original Message----- } From: Erasmus, Willem (WJ) {willem.erasmus-at-sasol.com} To: 'microscopy-at-msa.microscopy.com' {microscopy-at-Sparc5.Microscopy.Com}
Could someone please point me to an on-line resource (or previous letter to this group) that provides a protocol for embedding early post-implantation (E8-9.5) mouse embryos in paraffin?
We have tried embedding in gelatin as for cryostating, but the wax does not infiltrate properly.
Many thanks,
-Ron Goldstein -- ----------------------------------------------------------------------------- Dr. Ron Goldstein email goldst-at-mail.biu.ac.il Dept. Life Sciences Tel. 972-3-531-8216 Bar-Ilan Univ FAX 972-3-535-1824 52900 Ramat-Gan, ISRAEL http://www.biu.ac.il/NAT/ls/goldstein.html -----------------------------------------------------------------------------
} Hi- } } I am looking for the recipe for Dalton's Osmium Solution. I have a couple } of references, but the details of the recipe are different. Does anyone } have the citation of the original reference? Thank you very much! }
Dalton, A.J. Anatomical Record 121:281, 1955
-- ********************************************** 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 **********************************************
} } Subject: Re: Freeze fracture Need Help } } } } Dear freeze fracturers, } } } } We (BAL-TEC / Balzers) still manufacture double replica devices. Please } check our } } home page at bal-tec.com . For any questions please contact me off line. } } Best regards } } Alex Vogt } } BAL-TEC is a manufacturer of preparation equipment. } }
We would like to acquire STEM, SE, and BSE attachments for JEOL's JEM1200EX. Dealers welcome.
Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Electron Microscopy Facilities and Department of Biology University of California at San Diego
address: 1500 Bonner Hall, University of California at San Diego 9500 Gilman Drive, La Jolla, CA 92093-0368 telephone - lab: 8585342484 telephone - office: 8588223373 pager: 8586161420 email: mmm-at-biomail.ucsd.edu
Dear list members! I am servicing the SEM Hitachi S-800 at the Moscow State University. It is time now to replace the field emission cathode in this microscope. But unlike the substitution of heating cathodes the substitution of field emission ones is rather composite procedure. Could anybody prompt anything about any FEGSEM in this occasion? I shall take your advises with gratitude. Please you could reply by E-mail: antron-at-space.ru, or by fax (in USA): 603-761-3208.
Victor Sidorenko, ANTRON Co. Ltd., scientific service, Moscow, Russia.
Well, I think you do not have to shield the camera from the direct beam. In fact, I recorded a lot of EDs on our CCD (a 679 1k x 1k from Gatan) of biological 2D crystals (Brink & Chiu, J. Struct. Biol. (1994) 113:23). About the only thing ocurring when recording ED like this is overflow of the charge from the pixels with incident irradiation to neighbouring pixels (spike formation). At really high dose per ED (more than 1 e/angstrom^2 on the specimen, equivalent to about 10^7 e in the central beam) you may get some remnant of this spike in subsequent EDs. This imprint result from charge that gets deposited into the bulk of the silicon. However, if you are recording ED from inorganic materials, more than likely you should not have these problems. Besides, the imprint can easily be removed by raising the CCD temperature from -30C to RT. Alternatively, if you are afraid you may damage the CCD without using a beam stop (although we recorded well over 10,000 on our SSC without any signs of damage) you could inquire with your EM manufacturer about a custom built beam stop. I'm afraid though you may find the cost of such a beam stop prohibitively high.
Good luck with your efforts.
Jaap
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Rm. N420 Alkek Building, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Mon, 6 Sep 1999, Erasmus, Willem (WJ) wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Microscopists } } I have a problem obtaining diffraction patterns on our CCD camera. We have a } Philips CM200 TEM fitted with a Gatan Image Filter. Due to an extra } magnification factor introduced by the GIF the image on the TEM viewing } screen is much smaller than the image seen on the computer screen. Although } bothersome, this does not present too much of a problem in ordinary imaging. } } } However, when obtaining a diffraction pattern I find that one cannot shield } the CCD camera from the centre spot (transmitted beam) with the beam-stop } because the image of the beam-stop covers the diffraction pattern. The only } thing visible on the screen is the beam-stop. } } At the moment I am recording diffraction patterns on film, but is there a } way to use the CCD camera for this purpose ? } } }
The program is set for the meeting to be held at San Francisco State University on October 2nd. The program and abstracts are available at the meeting web page (http://online.sfsu.edu/~camicro/). Registration is also via this web page. Please join us.
greg antipa
Program Highlights below:
For program, flyers, and registration go to: http://online.sfsu.edu/~camicro/ or search camicro either at www.sfsu.edu or your favorite search engine
5th California Microscopy Colloquium The California State University & Northern California Society for Microscopy Saturday, October 2, 1999
Seven Hills Conference Center San Francisco State University
All fields of light and electron microscopy. Participants include scientists and students from academia and industry.
to include presentations by:
David Blake Life on Mars and life in the Mars Meteorite. Has the latter taken on a life of its own?
Jacek Gaertig Molecular and microscopic analysis of organelle assembly in Tetrahymena.
David Scharf Scanning Electron Microscopy in the 21st Century.
and 40 others
Register Online (http://online.sfsu.edu/~camicro/) Registration (From August 3rd to September 24th): Regular - $35, Student - $15 Registration includes both lunch and breaks Late Registration after September 24th & On Site: Regular - $35, Student - $15 For Late Registration, lunch may not be included dep. on the date of your registration.
or, in a pinch Greg Antipa Phone: (415) 338-2951 or EMail: antipa-at-sfsu.edu or FAX (415) 338-2295
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
The replacement of a Hitachi S-800 FE tip is fairly straightforward. A detailed procedure is in the service manual which you may or may not have.
This is not a trivial procedure but it is also not an overwhelming procedure: just tedious and not for someone who doe not have a "feel" for equipment. What I mean by a ""feel" for equipment' is someone who can work well with tools and instrumentation. If you have not don't try this at the lab.
Conversely, I would not be afraid of this either. A man put this together, a man can take it apart and repair it.
You will be working with an ultra-high vacuum system and consequently all the common sense procedures regarding cleanliness and attention to detail apply. Do not leave fingerprints, use several sets of lint-free gloves, rinse contaminated areas with reagent grade methanol, etc. I pre-clean the tools I use with acetone then methanol.
Make sure you have all the replacement parts including 6-inch conflat, new FE tip and springs.
As I can recall the procedure is as follows:
1. I usually ensure that I have all the replacement parts including a new FE tip, new springs, conflat gasket, new lint free gloves, aluminum foil, tweezers, 13mm socket with torque wrench. 2. I turn off all three the ion pump power supplies. 3. Remove the four piece shroud covering the ion pumps. Be careful not to short anything or turn the SEM power off then turn power back on to continue. 4. Turn off the " Display Power". 5. Remove the high voltage cable at the FE gun end. 6. Remove the three mu-metal shield from the column. 7. Clean and dust the FE surfaces. I use a dry paintbrush then follow the area using a lint-free cloth soaked in methanol. 8. Prepare a tabletop area to work on the gun. I dust the surface then follow with a lint-free cloth soaked in methanol. I then cover the area with aluminum foil. 9. Open the Ion-pump bypass valves (4) located to the right side of each of the ion pumps. CCW opens them. 10. Vent the specimen chamber. This will also vent the gun as the bypass valves are open. Venting with N2 helps. 11. Unbolt all 16 conflat bolts located at the top of the gun. Mark the gun chamber and gun with felt pen as a reference. 12. Refer to the operation manual: there are three silver sleeves and corresponding bolts in the toolkit. Looking at the gun studs from the top locate three studs that have the threads for the sleeve installation. remove the three studs and install the three sleeves and bolts. See manual for description as the manual has pictures that are difficult to descibe here. 13. CAREFULLY lift out the gun assembly. Lifting upward only. 14. Flip the gun assembly over and place on prepared table. 15. Use gloves at this point. Remove and discard old conflat. Replace with new, clean conflat opened from package. Cover open gun chamber with aluminum foil. 16. We are now working on the gun assembly. The FE tip has two pins:one insulated the other grounded to the assembly. Note the pin position. Loosen the two allen screw holding the pins. Remove the old FE tip. 17. The springs located on either side of the FE tip are now distorted and damaged. Remove the springs. Remove the pins located at the end of the springs. make sure the spring pins are not distorted. reshape them if necessary. Replace springs with the new ones ordered. When replacing the springs only screw on the springs with one turn at each end. In other words, do NOT screw the springs on as tight as they can. make the spring as straigjht as you can. 18. Re-install the FE tip observing the "polarity" - one side is insulated from the rest of the housing. Tighten the allen head screws after placing the FE tip in place. 19. Remove the aluminum foil from the gun chamber. 20. CAREFULLY re-install the gun ensuring that the pins align to the corresponding posts located down in the gun chamber. Not a easy task. 21. Using an ohm-meter, measure the resistance down in the gun assembly from the two side pins. These are the same two pins that you connect the "inner bake" cable to when baking the system.The resistance should be around 28 ohms. 22. If you have measured the proper resistance of 28 to 30 ohms, CONGRATULATIONS the worst is over. If not remove the gun and find out why. You may need to replace the pins again. 23. Remove the silver alignment bolt & sleeves. Tighten the gun bolts using a cross pattern and the proper torque listed in the instruction manual. Check resistance again. 24. Pump column down with bypass valves open. When pressure is better than 10-6, I bake the system (inner and outer) for at least 6 hours. 25. Close the three bypass valves along side of the ion pumps. The fourth bypass valve (bottom)is recommeded to be left open for some reason. 26. Follow the bake cycle procedure described in the instruction manual for a inner and out bake. 27. I usually bake the system three times before conditioning the gun for use. Gun conditioing is described in the instruction manual. 28. After the gun conditioning, the tip needs to be "blunted". Blunting is done by heavy flashing the tip three times in sucession. 29. The mu metal shields and the shrouds can be re-installed and the gun & column can be aligned. Gun & column alignment is also described in the instruction manual.
Easy wasn't it.
The gun replacement procedure should take 4 hours. Baking should take another three days for a total of about four days at best. This may seem like an overwhelming task but Hitachi has one of the best and easiest gun replacement procedures. Other designs require factory replacement and are the tip replacement cannot be done in the field.
Regards,
Earl Weltmer
Victor Sidorenko wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear list members! } I am servicing the SEM Hitachi S-800 at the Moscow State University. } It is time now to replace the field emission cathode in this } microscope. But unlike the substitution of heating cathodes the } substitution of field emission ones is rather composite procedure. } Could anybody prompt anything about any FEGSEM in this occasion? I } shall take your advises with gratitude. } Please you could reply by E-mail: antron-at-space.ru, } or by fax (in USA): 603-761-3208. } } Victor Sidorenko, ANTRON Co. Ltd., scientific service, Moscow, Russia.
I forgot to add the following step after step 24: activate the ion pumps.
Earl Weltmer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } The replacement of a Hitachi S-800 FE tip is fairly straightforward. A } detailed procedure is } in the service manual which you may or may not have. } } This is not a trivial procedure but it is also not an overwhelming } procedure: just tedious and not for } someone who doe not have a "feel" for equipment. What I mean by a ""feel" } for equipment' is someone who } can work well with tools and instrumentation. If you have not don't try this } at the lab. } } Conversely, I would not be afraid of this either. A man put this together, a } man can take it apart and repair it. } } You will be working with an ultra-high vacuum system and consequently all } the common sense procedures } regarding cleanliness and attention to detail apply. Do not leave } fingerprints, use several sets of lint-free gloves, } rinse contaminated areas with reagent grade methanol, etc. I pre-clean the } tools I use with acetone then methanol. } } Make sure you have all the replacement parts including 6-inch conflat, new } FE tip and springs. } } As I can recall the procedure is as follows: } } 1. I usually ensure that I have all the replacement parts including a new FE } tip, new springs, conflat gasket, } new lint free gloves, aluminum foil, tweezers, 13mm socket with torque } wrench. } 2. I turn off all three the ion pump power supplies. } 3. Remove the four piece shroud covering the ion pumps. Be careful not to } short anything or turn the SEM power off then turn power back on to } continue. } 4. Turn off the " Display Power". } 5. Remove the high voltage cable at the FE gun end. } 6. Remove the three mu-metal shield from the column. } 7. Clean and dust the FE surfaces. I use a dry paintbrush then follow the } area using a lint-free cloth soaked in methanol. } 8. Prepare a tabletop area to work on the gun. I dust the surface then } follow with a lint-free cloth soaked in methanol. } I then cover the area with aluminum foil. } 9. Open the Ion-pump bypass valves (4) located to the right side of each of } the ion pumps. CCW opens them. } 10. Vent the specimen chamber. This will also vent the gun as the bypass } valves are open. Venting with N2 helps. } 11. Unbolt all 16 conflat bolts located at the top of the gun. Mark the gun } chamber and gun with felt pen as a reference. } 12. Refer to the operation manual: there are three silver sleeves and } corresponding bolts in the toolkit. } Looking at the gun studs from the top locate three studs that have the } threads for the sleeve installation. } remove the three studs and install the three sleeves and bolts. See } manual for description as the manual } has pictures that are difficult to descibe here. } 13. CAREFULLY lift out the gun assembly. Lifting upward only. } 14. Flip the gun assembly over and place on prepared table. } 15. Use gloves at this point. Remove and discard old conflat. Replace with } new, clean conflat opened from package. } Cover open gun chamber with aluminum foil. } 16. We are now working on the gun assembly. The FE tip has two pins:one } insulated the other grounded to the assembly. } Note the pin position. Loosen the two allen screw holding the pins. } Remove the old FE tip. } 17. The springs located on either side of the FE tip are now distorted and } damaged. Remove the springs. Remove the pins } located at the end of the springs. make sure the spring pins are not } distorted. reshape them if necessary. Replace } springs with the new ones ordered. When replacing the springs only } screw on the springs with one turn at each end. } In other words, do NOT screw the springs on as tight as they can. make } the spring as straigjht as you can. } 18. Re-install the FE tip observing the "polarity" - one side is insulated } from the rest of the housing. Tighten the allen head screws } after placing the FE tip in place. } 19. Remove the aluminum foil from the gun chamber. } 20. CAREFULLY re-install the gun ensuring that the pins align to the } corresponding posts located down in the gun chamber. } Not a easy task. } 21. Using an ohm-meter, measure the resistance down in the gun assembly from } the two side pins. These are the same two pins } that you connect the "inner bake" cable to when baking the system.The } resistance should be around 28 ohms. } 22. If you have measured the proper resistance of 28 to 30 ohms, } CONGRATULATIONS the worst is over. } If not remove the gun and find out why. You may need to replace the } pins again. } 23. Remove the silver alignment bolt & sleeves. Tighten the gun bolts using } a cross pattern and the proper torque listed in the } instruction manual. Check resistance again. } 24. Pump column down with bypass valves open. When pressure is better than } 10-6, I bake the system (inner and outer) for at least 6 hours. } 25. Close the three bypass valves along side of the ion pumps. The fourth } bypass valve (bottom)is recommeded to be left open for some reason. } 26. Follow the bake cycle procedure described in the instruction manual for } a inner and out bake. } 27. I usually bake the system three times before conditioning the gun for } use. Gun conditioing is described in the instruction manual. } 28. After the gun conditioning, the tip needs to be "blunted". Blunting is } done by heavy flashing the tip three times in sucession. } 29. The mu metal shields and the shrouds can be re-installed and the gun & } column can be aligned. Gun & column alignment } is also described in the instruction manual. } } Easy wasn't it. } } The gun replacement procedure should take 4 hours. Baking should take } another three days for a total of about four days at best. } This may seem like an overwhelming task but Hitachi has one of the best and } easiest gun replacement procedures. } Other designs require factory replacement and are the tip replacement cannot } be done in the field. } } Regards, } } Earl Weltmer } } Victor Sidorenko wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear list members! } } I am servicing the SEM Hitachi S-800 at the Moscow State University. } } It is time now to replace the field emission cathode in this } } microscope. But unlike the substitution of heating cathodes the } } substitution of field emission ones is rather composite procedure. } } Could anybody prompt anything about any FEGSEM in this occasion? I } } shall take your advises with gratitude. } } Please you could reply by E-mail: antron-at-space.ru, } } or by fax (in USA): 603-761-3208. } } } } Victor Sidorenko, ANTRON Co. Ltd., scientific service, Moscow, Russia.
Yes, the danger of damaging the CAMERA itself is very low. I am not aware of any camera that is exposed to the beam directly. This would pose problems with sensitivity, damage to the CCD chip and many X-rays being recorded as well.
Cameras usually use some form of electron-light converter (Phosphor, YAG), then funnel the light to the CCD chip by use of a lens system or fiber-optic channel plate. Most CCD chips react to overexposure with light simply by "spilling" charge into the next pixels, which leads to "blooming". There are chips available that have anti-blooming guard rings around each pixel, but they are very expensive and they reduce sensitivity, as the guard ring area is not photo sensitive.
The part that suffers the most from exposure to high beam currents (as in the central diffraction spot) is probably the phosphor or YAG, which can be burned. Unlike during regular TEM, where the viewing screen is only used for viewing, a camera uses the same phosphor for viewing and recording images. You should try to avoid burning the phosphor to maintain good image quality later on.
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 ===================================
-----Original Message----- } From: Exchange Administrator Sent: Tuesday, September 07, 1999 12:08 AM To: Michael Bode
Well, I think you do not have to shield the camera from the direct beam. In fact, I recorded a lot of EDs on our CCD (a 679 1k x 1k from Gatan) of biological 2D crystals (Brink & Chiu, J. Struct. Biol. (1994) 113:23). About the only thing ocurring when recording ED like this is overflow of the charge from the pixels with incident irradiation to neighbouring pixels (spike formation). At really high dose per ED (more than 1 e/angstrom^2 on the specimen, equivalent to about 10^7 e in the central beam) you may get some remnant of this spike in subsequent EDs. This imprint result from charge that gets deposited into the bulk of the silicon. However, if you are recording ED from inorganic materials, more than likely you should not have these problems. Besides, the imprint can easily be removed by raising the CCD temperature from -30C to RT. Alternatively, if you are afraid you may damage the CCD without using a beam stop (although we recorded well over 10,000 on our SSC without any signs of damage) you could inquire with your EM manufacturer about a custom built beam stop. I'm afraid though you may find the cost of such a beam stop prohibitively high.
Good luck with your efforts.
Jaap
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Rm. N420 Alkek Building, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Mon, 6 Sep 1999, Erasmus, Willem (WJ) wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Microscopists } } I have a problem obtaining diffraction patterns on our CCD camera. We have a } Philips CM200 TEM fitted with a Gatan Image Filter. Due to an extra } magnification factor introduced by the GIF the image on the TEM viewing } screen is much smaller than the image seen on the computer screen. Although } bothersome, this does not present too much of a problem in ordinary imaging. } } } However, when obtaining a diffraction pattern I find that one cannot shield } the CCD camera from the centre spot (transmitted beam) with the beam-stop } because the image of the beam-stop covers the diffraction pattern. The only } thing visible on the screen is the beam-stop. } } At the moment I am recording diffraction patterns on film, but is there a } way to use the CCD camera for this purpose ? } } }
re: DV-23 Vacuum Gauge Tube for a Gatan Ion Miller
Hi all, Obviously I was not thinking very clearly last week, several people=20 pointed out that Duniway Stockroom was the perfect place to go for=20 this part! Many thanks for the help.
Here is a summary of the response I got (not attributions)
1. Kurt Lesker has the DV-23 for $58. Duniway Stockroom has an equivalent for $56.
2. Have you tried Lesker (www.lesker.com)? They are in western PA but usua= lly ship stuff out quickly.
3. You should give Duniway a call. They had a booth at M&M
4. Teledyne Hastings Chicago office: (804) 723-3925 As of 1994!
5. Yeah--call Teledyne Hastings-Raydist and ask for the name of your local dealer. When I did this, they had a $75 minimum order (2 tubes), so I had a spare....
6. Try Duniway Stock Room www.duniway.com 800-446-8811. Their=20 replacement for the DV-23 is part DST-023
7. Teledyne Hastings - Tel:757-723-6531, Fax:757-723-3925, E-mail :=20 Charles fulmer-at-teledyne.com
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"
We routinely record selected-area and parallel-nanopobe diffraction patterns with a bottom-mounted Gatan 1K x 1K CCD camera on our JEOL 2010F FEGTEM. Typical indicated camera lengths in the microscope are 15-20 cm to record spot or ring patterns for phase identification, and a bit less to get wide-angle patterns for orientation analysis from Kikuchi lines. The (12-bit) dynamic range of this camera is just about sufficient to capture the entire pattern intensity range up to the intense central beam with no beam stop. This usually takes a few adjustments of the exposure time so that the pattern center is just at or a little above saturation (at 14K to 16 K counts, depending whether you use dark current subtraction). To get the pattern intensity in range for recording times up to a few seconds, and of course to avoid damaging the YAG scintillator with the direct beam, you can reduce the intensity by choosing a small condenser aperture.
If you use dark current subtraction, be sure the beam deflection system of the microscope is set up to completely blank diffraction patterns. Otherwise, the automatically recorded dark current pattern will actually be a displaced diffraction pattern and the subtraction result will not be what you want. I usually use a continuous recording mode, and start recording with the camera blanked by the microscope viewing screen. In this mode, you can dynamically adjust the exposure time, set the contrast, and make other corrections before recording the final pattern.
Finally, lens hysteresis in the microscope can have a large effect on pattern (camera constant) calibration at these small camera lengths.
Larry
Larry Thomas Battelle, Pacific Northwest Laboratories Richland, WA
Larry.Thomas-at-pnl.gov (509) 372-0793
---------- From: Michael Bode Sent: Tuesday, September 7, 1999 6:50 AM To: 'Microscopy-at-MSA.Microscopy.Com' Subject: RE: Diff patterns
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Yes, the danger of damaging the CAMERA itself is very low. I am not aware of any camera that is exposed to the beam directly. This would pose problems with sensitivity, damage to the CCD chip and many X-rays being recorded as well.
Cameras usually use some form of electron-light converter (Phosphor, YAG), then funnel the light to the CCD chip by use of a lens system or fiber-optic channel plate. Most CCD chips react to overexposure with light simply by "spilling" charge into the next pixels, which leads to "blooming". There are chips available that have anti-blooming guard rings around each pixel, but they are very expensive and they reduce sensitivity, as the guard ring area is not photo sensitive.
The part that suffers the most from exposure to high beam currents (as in the central diffraction spot) is probably the phosphor or YAG, which can be burned. Unlike during regular TEM, where the viewing screen is only used for viewing, a camera uses the same phosphor for viewing and recording images. You should try to avoid burning the phosphor to maintain good image quality later on.
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 ===================================
-----Original Message----- } From: Exchange Administrator Sent: Tuesday, September 07, 1999 12:08 AM To: Michael Bode Subject: FW: Diff patterns
} ---------- } From: "brink-at-tiger.3dem.bioch.bcm.tmc.edu"-at-sparc5.microscopy.com[SMTP:"BRINK-at-T IGER.3DEM.BIOCH.BCM.TMC.EDU"-at-SPARC5.MICROSCOPY.COM] } Sent: Monday, September 06, 1999 8:24:38 PM } To: Erasmus, Willem (WJ) } Cc: 'microscopy-at-msa.microscopy.com' } Subject: Re: Diff patterns } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Well, I think you do not have to shield the camera from the direct beam. In fact, I recorded a lot of EDs on our CCD (a 679 1k x 1k from Gatan) of biological 2D crystals (Brink & Chiu, J. Struct. Biol. (1994) 113:23). About the only thing ocurring when recording ED like this is overflow of the charge from the pixels with incident irradiation to neighbouring pixels (spike formation). At really high dose per ED (more than 1 e/angstrom^2 on the specimen, equivalent to about 10^7 e in the central beam) you may get some remnant of this spike in subsequent EDs. This imprint result from charge that gets deposited into the bulk of the silicon. However, if you are recording ED from inorganic materials, more than likely you should not have these problems. Besides, the imprint can easily be removed by raising the CCD temperature from -30C to RT. Alternatively, if you are afraid you may damage the CCD without using a beam stop (although we recorded well over 10,000 on our SSC without any signs of damage) you could inquire with your EM manufacturer about a custom built beam stop. I'm afraid though you may find the cost of such a beam stop prohibitively high.
Good luck with your efforts.
Jaap
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Rm. N420 Alkek Building, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Mon, 6 Sep 1999, Erasmus, Willem (WJ) wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Microscopists } } I have a problem obtaining diffraction patterns on our CCD camera. We have a } Philips CM200 TEM fitted with a Gatan Image Filter. Due to an extra } magnification factor introduced by the GIF the image on the TEM viewing } screen is much smaller than the image seen on the computer screen. Although } bothersome, this does not present too much of a problem in ordinary imaging. } } } However, when obtaining a diffraction pattern I find that one cannot shield } the CCD camera from the centre spot (transmitted beam) with the beam-stop } because the image of the beam-stop covers the diffraction pattern. The only } thing visible on the screen is the beam-stop. } } At the moment I am recording diffraction patterns on film, but is there a } way to use the CCD camera for this purpose ? } } }
A few years ago I saw at an MSA meeting a poster detailing how a low vibration room for TEM's had been designed and constructed as part of a new building. It involved the pouring of a concrete slab uniquely for a TEM and separate from the walls to minimize vibration. I thought I had a copy of the information provided by the poster but cannot seem to find it. Does anyone know:
1) Who authored the poster and a contact number? 2) Where I might find MSA poster information archived? 3) Any other information related to building a low vibration room as part of a new building?
Thanks in advance for your time and help.
Sincerely,
Mick Thomas ------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
At 04:20 PM 9/7/99 -0700, Mick Thomas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
A paper covering the installation concerns, including vibration, for a FEG-TEM can be found in the proceedings from the MRS Spring 1999 symposium "Electron Microscopy of Semiconducting Materials and ULSI Devices."
C.J.D. Hetherington, et.al., "Installing and Operating FEGTEM's," Mater. Res. Soc. Proc. Vol. 523, ed. by C. Hayzelden, C. Hetherington, and F. Ross, pp. 171-6.
Philip L. Flaitz IBM Analytical Services, Hopewell Junction, NY http://www.chips.ibm.com/services/asg Ph.......(914) 892-3094, FAX -2003 flaitz-at-us.ibm.com
Mick Thomas {mgt3-at-msc.cornell.edu} on 09/07/99 07:20:44 PM
To: microscopy-at-sparc5.microscopy.com cc:
Fellow microscopists,
A few years ago I saw at an MSA meeting a poster detailing how a low vibration room for TEM's had been designed and constructed as part of a new building. It involved the pouring of a concrete slab uniquely for a TEM and separate from the walls to minimize vibration. I thought I had a copy of the information provided by the poster but cannot seem to find it. Does anyone know:
1) Who authored the poster and a contact number? 2) Where I might find MSA poster information archived? 3) Any other information related to building a low vibration room as part of a new building?
Thanks in advance for your time and help.
Sincerely,
Mick Thomas ------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
Mick, The poster you saw was probably: "Design and implementation of a site for a one-Ångstrom TEM", John H. Turner, Michael A. O'Keefe and Robert Mueller, in 55th Ann. Proc. MSA, Cleveland, Ohio (1997) 1177-1178. Contact me or John turner if you want more details. -Mike O'Keefe p.s. see http://ncem.lbl.gov/frames/oam.htm for images obtained using the microscope in that room.
Mick Thomas {mgt3-at-msc.cornell.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} of the information provided by the poster but cannot seem to find it. Does } anyone know: } } 1) Who authored the poster and a contact number? } 2) Where I might find MSA poster information archived? } 3) Any other information related to building a low vibration room as part } of a new building? } } Thanks in advance for your time and help. } } Sincerely, } } Mick Thomas } ------------------------- } Mick Thomas } UHV-STEM Laboratory } E-1 Clark Hall } Cornell University } Ithaca, NY 14853 } } Phone: 607-255-0650 } Fax: 607-255-7658 } e-mail: mgt3-at-msc.cornell.edu } }
Mick Thomas wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Fellow microscopists, } } A few years ago I saw at an MSA meeting a poster detailing how a low } vibration room for TEM's had been designed and constructed as part of a new } building. It involved the pouring of a concrete slab uniquely for a TEM } and separate from the walls to minimize vibration. I thought I had a copy } of the information provided by the poster but cannot seem to find it. Does } anyone know: } } 1) Who authored the poster and a contact number? } 2) Where I might find MSA poster information archived? } 3) Any other information related to building a low vibration room as part } of a new building? } } Thanks in advance for your time and help. } } Sincerely, } } Mick Thomas } ------------------------- } Mick Thomas } UHV-STEM Laboratory } E-1 Clark Hall } Cornell University } Ithaca, NY 14853 } } Phone: 607-255-0650 } Fax: 607-255-7658 } e-mail: mgt3-at-msc.cornell.edu
Mick,
Contact Black Engineering Company (shock, vibration and noise isolation): (800)747-2523 or (404)578-0999. It may or may not be the one on the poster, yet they do exactly what you are looking for. Cheers.
Does anyone on the list have any idea or experience in measuring the volume of fingerprints? Many Thanks in advance Dr. G.Sanders -------------------------------------------------------------------------------- -----------------------Dr. Giles SandersZeneca / SmithKline Beecham Centre for Analytical SciencesChemistry DepartmentImperial College of Science, Technology and MedicineLondonSW7 2AY
Dear William: It is so nice and comforting to see a gentleman who expresses concerns over such important matters. I wish I had you as an instructor. Thank you. Now to respond...I was pregnant with my first child while working as an EMst and left work in my 11th month. This was also the time when everyone was hopping on the bandwagon in AIDS research and I found myself working with blood serum etc. trying to image this virus. As a result, I gave birth to a healthy baby girl and experienced no problems. She is now a beautiful 15-year old. If all the precautions are taken: nitrile gloves, using an exhaust hood pulling at least 100 feet per sec., disposable lab coats, goggles, etc., there should be no problem. A whole unit ought to be devoted to these issues for all students when learning EM methods. Also if the scopes are monitored for X ray leakage and the meter shows only background readings, then don't worry. My only concern is when changing the filaments...maybe some residual x ray exposure. But I don't see any big problem. Just monitor this woman to make sure if she is using all the precautionary steps and she should be fine....and have fun!!!!
Regards, 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
What exactly do you mean by "volume of fingerprints"?
Is that the volume of the residue left by the finger? Is it the volume of the finger that left the imprint? Do you mean the area of the fingerprint?
I am by no means an expert on fingerprints, but unless you have some additional information, I would guess that the extraction of a volume from 2-dimensional data (such as a print) is very difficult. On the other hand, if you do have additional information that applies to all fingers of all people (perhaps something as an "internal finger pressure"), and you know the force that was used by the finger that left the imprint, you might be able to calculate the volume of the finger from the area of the fingerprint.
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: Giles Sanders[SMTP:G.SANDERS-at-IC.AC.UK] } Sent: Wednesday, September 08, 1999 1:06:46 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Fingerprints } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone on the list have any idea or experience in measuring the volume of fingerprints? Many Thanks in advance Dr. G.Sanders ------------------------------------------------------------------------ -------- -----------------------Dr. Giles SandersZeneca / SmithKline Beecham Centre for Analytical SciencesChemistry DepartmentImperial College of Science, Technology and MedicineLondonSW7 2AY
We are looking for a conductive(minimize charging) epoxy to backfill cracks and voids in metals to minimize water, solvent and etchant leakage after polishing.
It is most distressing to attempt to do probe analysis on a leaking sample.
Michael: the solution is often simple depending on the level of accuracy = required. If one knows the density of fingerprint oil then one can = "weigh" several fingerprints and average out the volume. Also one could = acetone clean the finger free of natural oil and ink up the finger with = fingerprinting ink (which you could more easily determine the density of) = transfer to previously weighed gelatin paper and weigh a number of prints = (by subtraction) on a high quality analytical balance and determine the = volume since volume equals weight (actually mass) divided by density. = thanks
I am in need of updated information or websites on the transmission electron microscope and the scanning-tunneling microscope. Any help would be appreciated. Thank you.
Has anyone had success in performing LM autoradiography on GMA (JB4 gly= col methacrylate) sections? I'm having trouble keeping the emulsion on the=
sections during the wash step, after the slides have been fixed in photographic fixer. The area of emulsion overlying the sections only p= uckers and peels off within 3 minutes into the wash step. So far I've tried c= asting Kodak NTB2 undiluted or Ilford K.5 (dil=3D1:1) emulsions on clean slid= es with 4 um GMA sections, on gelatin subbed and unsubbed slides.
To develop autoradiograms I'm using:
D-19 developer (dil=3D1:1) ------4 minutes Stop bath (0.5% acetic acid in dist water)-----30 sec 30% sodium thiosulfate fixer (no hardener) -----5 minutes distilled water wash (slides vertical in vessel)----15 minutes
Next, I will be trying diluted Kodak NTB2 emulsion (dil=3D 1:1), elimin= ating the acid in the stop bath, and switching to a fixer with hardener added= (eg. Kodak fixer).. maybe a few other things, but I'm running short on time.=
Any suggestions or references would be greatly appreciated.
Thanks in advance :)
Figen Seiler Microscopist Abbott Laboratories Department of Microscopy & Microanalysis E-mail: figen.a.seiler-at-abbott.com
A soulution that works as well is to cut a hole in the floor and drive wooden piles in the hole. It only work on lower floors. but it is better a killing vibrations than concrete.
Gordon Couger 624 Cheyenne Stillwater, OK 74075 405 624 2855 GMT - 6:00 -----Original Message----- } From: Vitaly Feingold {vitalylazar-at-worldnet.att.net} To: Mick Thomas {mgt3-at-msc.cornell.edu} Cc: microscopy-at-Sparc5.Microscopy.Com {microscopy-at-Sparc5.Microscopy.Com}
A soulution that works as well is to cut a hole in the floor and drive wooden piles in the hole. It only work on lower floors. but it is better a killing vibrations than concrete.
Gordon Couger 624 Cheyenne Stillwater, OK 74075 405 624 2855 GMT - 6:00 -----Original Message----- } From: Vitaly Feingold {vitalylazar-at-worldnet.att.net} To: Mick Thomas {mgt3-at-msc.cornell.edu} Cc: microscopy-at-Sparc5.Microscopy.Com {microscopy-at-Sparc5.Microscopy.Com}
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This is a common problem when examining polished sections in the SEM. Due to shrinkage, there is often a gap between the edge of the sample and the plastic. This depends on the type of resin used and the speed of cure. Slower is better. The next important step is adequate ultrasonic cleaning in iso-propanol to remove all traces of the polishing compound. If the gap is very large, we infill with some more of the same epoxy and repeat the process, finally drying the solvent out with a hot air gun. Of course, we give a flash carbon coat and use carbon dag to give a good conducting path.
This seems to work for us, good luck!
Barry
} -----Original Message----- } From: Giles, Bill [SMTP:William.Giles-at-TIMET.com] } Sent: Wednesday, September 08, 1999 5:51 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Conductive epoxy for SEM/microanalysis use? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings, } } We are looking for a conductive(minimize charging) epoxy to backfill } cracks } and voids in metals to minimize water, solvent and etchant leakage after } polishing. } } It is most distressing to attempt to do probe analysis on a leaking } sample. } } Someone out there must have a solution!! } } Any suggestions? } } Bill Giles } TIMET }
Kelvin Conference Centre=20 University of Glasgow Wednesday 10 November 1999=20
The format for this successful series of one day meetings remains largely u= nchanged, with a programme of talks=20 interspersed with time for posters and the Trade Exhibition.
The following topics will be presented by key-note speakers:
(a) Microscopy of the Underworld; Looking at the adhesive surface of cells by techniques including Interferen= ce reflection, TIRFM, Forster Energy transfer and Surface Plasmon Resonance Microscopy.=20 Prof. Adam Curtis, Institute of Biomedical and Life Sciences, University of= Glasgow=20
(b) Using electrons to explore materials. Prof. Alan Craven, Department of = Physics and Astronomy, University of Glasgow
(c) Egg Tapping; Biomineralisation studied by SEM, Acoustic Resonance and L= aser Scanning Microscopy. Prof. Sally Solomon, Veterinary Anatomy, University of Glasgow
(d) Digital Information Acquisition in the TEM. Dr Timon Fliervoet, Phillips Electron Optics Applications Laboratory, Eindh= oven =20 A special feature this year will be short, contributed, prize talks by youn= g "first time" speakers.
There will be sponsored Prizes and these enjoyable, interesting and useful = meetings provide an opportunity to meet and discuss ideas and tips with microscopists in all fields.
The Conference Centre provides interspersed Meeting, Exhibition, Poster, Bu= ffet and Bar facilities and there is ample car parking. The cost per head is =A320, including the buffet lunch, coffee and tea. Subsidised buses= will run from Aberdeen, Edinburgh and Glasgow (contact the local User Group). This one day meeting is CPD accredited.
Web site: http://www.abdn.ac.uk/~nhi691/smg99.htm has abstracts form=20 the following meetings: 1998 Dundee Meeting=20 1997 Dunblane Meeting=20 1996 Aberdeen Meeting=20 =20
---------------------- Kevin Mackenzie Electron Microscope unit Dept Zoology University of Aberdeen Tillydrone avenue Aberdeen AB24 2TZ ----------------- Tel 01224 272847 Fax 01224 272396 email k.s.mackenzie-at-abdn.ac.uk
I need to collect the general and special ideas and tips on how to prepare the multilayered thin film TEM specimens (Both plan view and cross sections), especially for the magnetic films.
Thank you very much for your help !
You can reply the message to: c.jiao-at-bham.ac.uk
Many EM immuno protocols using colloidal gold on grid mounted thin sections employ a 1% glutaraldehyde step after the grid is incubated in the secondary gold-conjugated antibody and washed. Does anybody know whether this step is really necessary (i.e., is there a paper that compares fix vs no fix after labeling). I would have thought the affinity of the antibody would have precluded the need for this step. Comments welcome.
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 Bill, The last time I looked at buying conductive mounting epoxy, most suppliers listed a hot-press conductive resin, but not cold curing. I found a cold-curing, copper filled resin called Technovit 5000, from Germany. Kulzer: is the name of the company that makes it, but I ordered it from Energy Beam Sciences. It is a bit viscous to fill cracks, though. At 10:51 AM 9/8/99 -0600, you wrote: } } Greetings, } } We are looking for a conductive(minimize charging) epoxy to backfill cracks } and voids in metals to minimize water, solvent and etchant leakage after } polishing. } } It is most distressing to attempt to do probe analysis on a leaking sample. } } Someone out there must have a solution!! } } Any suggestions? } } Bill Giles } TIMET } 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
I have a student in my introductory electron microscopy course who has an implanted microprocessor controlled drug delivery pump. I am at the point where the "field trips" to the lab will start soon, and he asked me whether being close to the working microscopes (TEM at 75 kV, SEM at 5 kV) could interfere with his device. Although my initial thought was "I don't think so", I would like to make sure. One of the main things I've learned from reading the discussions that take place on this listserve is that there's a whole lot of physics out there that I don't know enough about.
I've posed this question to our environmental health and safety department (no answer yet), looked through some EM lab safety chapters in books, and checked the most excellent "Tips and Tricks of Microscopy" website to see if this has been discussed before. So far I haven't found any information.
I'd appreciate hearing from anyone who has insight into this kind of situation.
Thanks in advance,
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
If we can answer any questions, give us a call at 800-440-0311.
Hope this is helpful.
Elinor Solit The Microscope Book
On Wed, 8 Sep 1999 Joelis99-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 } -----------------------------------------------------------------------. } } } I am in need of updated information or websites on the transmission electron } microscope and the scanning-tunneling microscope. Any help would be } appreciated. } Thank you. } } Joel Simons } joelis99-at-aol.com }
looks like you want to measure the volume of the actual fingerprint.
I could think of 4 ways to do that:
Use an AFM type of instrument. This may or may not work, depending on the characteristics of the fingerprint oil. It also may have a field of view that is too restricted. But if you can use such an instrument, it could give you the volume immediately.
Use a confocal microscope to get at the height data. If you can do that, it should be fairly easy to extract the volume. You may have problems if the fingerprint oil is transparent to the frequency used, or if the "height" of the fingerprint is below the resolution limit of the instrument.
Use a standard light microscope and take pictures at different focus settings. If the fingerprints have enough depth, you can reconstruct the surface from these images and get the volume that way.
Use a Stereo pair and calculate the 3rd dimension. This requires that the depth of the fingerprint is considerable. I would guess, that this does not work, but may be worth a try.
We have software to analyze all these measurements. Please contact me off-line if you want more info.
Thanks.
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: Robert Mixon[SMTP:MIXONR-at-OHSU.EDU] } Sent: Wednesday, September 08, 1999 3:35:35 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: RE: fingerprints } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Michael: the solution is often simple depending on the level of accuracy required. If one knows the density of fingerprint oil then one can "weigh" several fingerprints and average out the volume. Also one could acetone clean the finger free of natural oil and ink up the finger with fingerprinting ink (which you could more easily determine the density of) transfer to previously weighed gelatin paper and weigh a number of prints (by subtraction) on a high quality analytical balance and determine the volume since volume equals weight (actually mass) divided by density. thanks
No great theory here but, by way of anecdotal input, this. In the past I've "borrowed" a TA for my TEM course from our MSM department. He has an insulin pump that, I believe, has a microprocessor control. He hangs around all sorts of TEM and SEM equipment for extend periods of time with no apparent (at least to my untrained eye) ill effects. He may even be lurking out there now and reply. cheers, John
John Heckman TEM Supervisor MSU Center for Electron Optics
Has anyone ever done in situ hybridization in cultured cells using 96-well culture plates? If so, do you have a favorite procedure that is followed? Any help would be greatly appreciated. Thanks in advance. Linda Chicoine Clinton, CT lchicoine-at-snet.net
I am soliciting contributors (or names of potential contributors) for a symposium for the Microscopy & Microanalysis meeting to be held on August 13-17, 2000 in Philadelphia, Pa.
Talks may range in length from 15 to 45 minutes.
A description of the symposium follows.
SYMPOSIUM: EXTREME ORGANISMS
This symposium will deal with organisms that represent extremes, as for example: the ability to grow in extreme environments, having extreme virulence or invasiveness, or being extremely difficult to visualize using conventional preparatory procedures. Hopefully, the participants shall describe some of the unique features of extreme organisms that give rise to these capabilities. Of particular interest shall be talks dealing with organisms able to grow in challenging environments such as high salt, high or low temperatures, high or low pressure and highly toxic conditions. In addition, speakers are invited to discuss organisms that are extremely virulent or invasive, as would be the case with certain pathogens. In this instance, whenever possible, speakers may identify the features of the organisms giving rise to this capability. Finally, extreme organisms are often difficult to visualize using standard preparatory procedures. Papers describing procedures to prepare the specimens for visualization would be germane to this symposium.
Please respond directly to me with the name of the individual and possible topic.
John Bozzola
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
Just to clarify my origninal question, by volume of the fingerprint I meant the volume of material left by the fingerprint, not a volume of imprint. Thanks in adavnce and for those who have already replied. -------------------------------------------------------------------------------- -----------------------Dr. Giles SandersZeneca / SmithKline Beecham Centre for Analytical SciencesChemistry DepartmentImperial College of Science, Technology and MedicineLondonSW7 2AY
Recently I got very interesting results on PET/PMMA extrudate morphology(10 ~ 1 wt% of PMMA and PET was crystalline). Instrument: Hitachi S800 SEM Sampling method: hand fracturing or knife cutting under liquid nitrogen temperature. Results: Unexpectedly, hand fracturing sample has little matrix burrs while knife cutting sample has many. This is strange because it is usually think cutting is better way than fracture. Any suggestions? Thank inadvance for your help. Dr. Zeng JijunVisiting Scholar of Toray Home Address: Nakatogari 734-2-5 Nagaizumi-cho, Sunto-gun Shizuoka, 411-0942 Japan Tel: +81-559-884601(H)E-mail: sentoray-at-izu.co.jpPersonal Homepage:http://www.geocities.com/ResearchTriangle/Forum/1786
There are only five openings left for the "Cryoultramicrotomy and Immunolabelling Workshop" being held at Harvard Medical School, October 5-7, 1999. The deadline for registration has been extended until 9/13/99 for those needing hotel accommodations and until 9/28/99 for those who do not. For further information, please contact Sonja White (swhite-at-ebsciences.com) {mailto:swhite-at-ebsciences.com)} .
Best regards, Steven Slap .******************************************* Energy Beam Sciences, Inc. The Laboratory Microwave Company Adding Brilliance to Your Vision http://www.ebsciences.com {http://www.ebsciences.com} *******************************************
Hello everyone I have just started a new season of "Small Wonders" on Discovery Channel. This is a picture puzzle, usually taken on an SEM, with clues. You have to guess the answer and win a prize!
It is only shown on TV in Canada as it is on -at-Discovery Canada, every Monday evening 8-9pm (Pacific time). It is usually the last item on the show. If you are not in Canada, you can still participate in the competition from the website. If you want to see the show, you can download "RealPlayer" onto your computer.
-at-Discovery website: www.exn.ca/-at-discovery.ca Click on the Small Wonders icon down the right hand side
If you go to the Archives at the bottom of the Small Wonders page, you can see some of the images from last year. Challenge: see if you can tell what the picture is before you see the answer!
Last year was fun and I hope to have more fun this year. If you have any ideas, please let me know off line - we wouldn't want to give all the answers away would we?! 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
Mary & Bill: Conductive silver epoxy in small twin tubes is available from ProSciTech and I expect some other "EM Suppliers". Its cold curing too. 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 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, September 10, 1999 2:36 AM, Mary Mager [SMTP:mager-at-interchange.ubc.ca] wrote: } } Dear Bill, } The last time I looked at buying conductive mounting epoxy, most suppliers } listed a hot-press conductive resin, but not cold curing. I found a } cold-curing, copper filled resin called Technovit 5000, from Germany. } Kulzer: is the name of the company that makes it, but I ordered it from } Energy Beam Sciences. It is a bit viscous to fill cracks, though. } At 10:51 AM 9/8/99 -0600, you wrote: } } } } Greetings, } } } } We are looking for a conductive(minimize charging) epoxy to backfill cracks } } and voids in metals to minimize water, solvent and etchant leakage after } } polishing. } } } } It is most distressing to attempt to do probe analysis on a leaking sample. } } } } Someone out there must have a solution!! } } } } Any suggestions? } } } } Bill Giles } } TIMET } } } 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 }
I have an Eico sputter coater with a gold target. I used to coat my ceramic samples with the coater for several years, and did not have any problem. A couple of weeks ago, I tried to coat biological sample which was a part of an animal tongue The sample was dried thoroughly, accroding to the person who prepared the sample, but the other sample prep process he used was unknown, unfortunately. After the sputter coating, something went wrong, and I could not coat my ceramic samples well any more. I checked the gold target with an optical microscope, and I found that green particles or at least particle-like stuffs were inside the pits or grooves on the gold target. It seems that my ceramic samples were not coated with gold but with the green stuffs. And also, I found the surface of the gold target was very rough and the color was a little reddish, comparing with the other side of the target, but I was not sure if this resulted from the coating procedure of biological sample or from the long time wear. My question is how I can clean the gold target. Have any of you exprienced this kind of problem before? Any comment is appreciated.
Jondo Yun Electron Microscopy Laboratory Center for the Instrumental Analysis Kyungnam University Masan, Korea
I would dis-assemble, clean and rinse with acetone, for the parts that ca= n take acetone (non-plastics), then follow with an alcohol rinse. Changing the r= otary pump oil would not hurt either.
Earl Weltmer
=C0=B1=C1=B8=B5=B5 wrote:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } } Dear all } } I have an Eico sputter coater with a gold target. I used to coat my } ceramic samples with the coater for several years, and did not have any } problem. } A couple of weeks ago, I tried to coat biological sample which was a pa= rt of } an animal tongue The sample was dried thoroughly, accroding to the pers= on } who prepared the sample, but the other sample prep process he used was } unknown, unfortunately. After the sputter coating, something went wrong= , and } I could not coat my ceramic samples well any more. } I checked the gold target with an optical microscope, and I found that = green } particles or at least particle-like stuffs were inside the pits or groo= ves } on the gold target. It seems that my ceramic samples were not coated wi= th } gold but with the green stuffs. } And also, I found the surface of the gold target was very rough and the } color was a little reddish, comparing with the other side of the target= , but } I was not sure if this resulted from the coating procedure of biologica= l } sample or from the long time wear. } My question is how I can clean the gold target. Have any of you exprien= ced } this kind of problem before? } Any comment is appreciated. } } Jondo Yun } Electron Microscopy Laboratory } Center for the Instrumental Analysis } Kyungnam University } Masan, Korea } } jdyun-at-hanma.kyungnam.ac.kr
Dear Listers, I'm inquiring about the quality of the carbon coating one can obtain using the table top coaters equipped with a mechanical pump. Also, has anyone has upgraded such a system with a turbomolecular pump? Rosemary
My rough pumped (only) carbon yarn evaporator works quite well for SEM specimens. I have no experience using it for TEM specimens, but believe the turbo type is better suited for that application.
Woody White
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Dear Listers, I'm inquiring about the quality of the carbon coating one can obtain using the table top coaters equipped with a mechanical pump. Also, has anyone has upgraded such a system with a turbomolecular pump? Rosemary
We have tried both ways many times due to trouble shooting when the labeling doesn't work. For us it doesn't make any difference. If the primary stuck, the secondary seemed to stick and it didn't make any change in label frequency or efficiency whether it is fixed after the secondary or not.
Bob Morphology Core Seattle
On Thu, 9 Sep 1999, Tom Phillips wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Many EM immuno protocols using colloidal gold on grid mounted thin } sections employ a 1% glutaraldehyde step after the grid is incubated } in the secondary gold-conjugated antibody and washed. Does anybody } know whether this step is really necessary (i.e., is there a paper } that compares fix vs no fix after labeling). I would have thought } the affinity of the antibody would have precluded the need for this } step. Comments welcome. } } 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 Gaetan, The S-570 cahmber is circular, with a flat front and is about 11.5 inches O.D. and 10 inches I.D. At 09:01 PM 9/9/99 -0500, you wrote:
} } Hi everybody, } Is there anybody who could give me the standard size of the chamber of the } Hitachi S-570? } } Thanks, } } Ga=EBtan } 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
Dear Jondo, I have wiped my gold target with acetone on a tissue to clean it. Sounds like something outgassed from the sample. Slimy biologists.;-) At 04:04 PM 9/10/99 +0900, you wrote:
} } Dear all } } I have an Eico sputter coater with a gold target. I used to coat my } ceramic samples with the coater for several years, and did not have any } problem. } A couple of weeks ago, I tried to coat biological sample which was a part of } an animal tongue The sample was dried thoroughly, accroding to the person } who prepared the sample, but the other sample prep process he used was } unknown, unfortunately. After the sputter coating, something went wrong, and } I could not coat my ceramic samples well any more. } I checked the gold target with an optical microscope, and I found that green } particles or at least particle-like stuffs were inside the pits or grooves } on the gold target. It seems that my ceramic samples were not coated with } gold but with the green stuffs. } And also, I found the surface of the gold target was very rough and the } color was a little reddish, comparing with the other side of the target, but } I was not sure if this resulted from the coating procedure of biological } sample or from the long time wear. } My question is how I can clean the gold target. Have any of you exprienced } this kind of problem before? } Any comment is appreciated. } } Jondo Yun } Electron Microscopy Laboratory } Center for the Instrumental Analysis } Kyungnam University } Masan, Korea } } jdyun-at-hanma.kyungnam.ac.kr } 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
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Reply to: RE: colloidal gold staining Dear Tom,
The glutaraldehyde step has been used for two primary purposes. The first = (and original) was to cross-link the gold probe to the primary antibody = and thus keep the signal in its correct location. However, binding of = antibodies or protein A to other antibodies is reasonably stable so many = protocols often omit this crosslinking step. The sections are often dried = soon after fixation which may also keep the gold in place. Other than the = original papers where the use of glutaraldehyde is described I don't know = of any papers comparing signal with and without this step.
The second use of 1% glutaraldehdye is when sections are being labeled with= two antibodies and two sizes of protein A gold. The usual protocol for = this in sequential labeling is: 1st antibody First gold probe (usually the smaller size) Blocking 2nd antibody Second gold probe.
To prevent the second gold probe from binding to the first antibody = earlier papers suggested the use of free protein A between the first gold = probe and the second antibody (the "Blocking" step). This effectively = covered any free protein A-binding sites that could confuse the double = labeling results. More recently, a 1% glutaraldehyde step has been used = to replace the free protein A-step. It seems to work better than the = protein A and is generally accepted to be a usable protocol for multiple = labeling. = = The reference for this is: van Genderen, I.L. van Meer G. Slot J.W. Geuze H.J. & Voorhout W.F. 1991. = Subcellular localization of Forssman glycolipid in epithelial MDCK cells = by immuno-electronmicroscopy after freeze-substitution. J. Cell Biol. 115:= 1009-1019. =
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 213 273 8026 213 413 6739 (fax) pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Tom Phillips wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
by attila.stevens-tech.edu (8.9.3/8.9.3/7) with ESMTP id NAA02448 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 10 Sep 1999 13:53:50 -0400 (EDT) Message-ID: {37D9468D.B9519533-at-stevens-tech.edu}
Greetings,
Recently I have embedded biological polymer fibers in epoxy to section for TEM samples. Unfortunately, the fibers with very small diameters dissolved in the epoxy. The fibers are used for applications requiring the polymer to be absorbed in the human body. Therefore, they also dissolve in water. Does anyone have any suggestions for either alternate methods to obtain a TEM sample or ways to alleviate this problem?
I don't know where to start, so any advice would be very useful.
Thanks,
Jennifer Taylor Ph.D. candidate Stevens Institute of Technology Hoboken, New Jersey 07030
I have a former student who is working with hyaluronic acid microspheres. These microspheres are clearly visible under the stereomicroscope, however, the desire is to view them using SEM. The problem is that when they are applied to a stub and allowed to air dry they disrupt in some way leaving a film layer behind. They wish to visualize the intact microspheres. In addition, they might also want to try observing them using a TEM (having worked with embedding and sectioning spheres/bubbles of surfactant from the lungs I realize how difficult this might be). If they could be embedded and sectioned what stain(s) might be employed?
The following is a description of how these microspheres are prepared. Any assistance with even just a SEM protocol would be greatly appreciated!
HA Preparation: " A hyaluronic acid (HA)solution and an organic oil (with a small amount of emulsifying agent) are mixed thoroughly until an emulsion forms. ONce the emulsion forms, polymerizing and cross-linking chemicals are added to cross-link the HA. The mixture is then centrifuged and the oil layer is discarded. The HA microsphere pellet is then washed several times with isopropanol and resuspended in a minimum amount of distilled water and then lyophilized."
Regards,
Steve
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
I have a question I have not been able to locate the answer to anywhere. I run a clinical EM lab here at the University of Nebraska and we have recently become associated with the College of Anatomic Pathologists (CAP) and I am having trouble locating any set regulations or even guidelines as to the duration we should retain records, as well as specimen blocks, thick section slides and EM photos. Does anyone have any info to get me headed in the right direction?
Thank you,
Doug Rennie Coodinator-EM Facility University of Nebraska Medical Center Omaha, Nebraska.
We are familiar with vacuum impregnation and the usual thermosetting products.
One of the problem areas that we have with using "normal" epoxy and carbon coating is that we primarily are trying to quantify nitrogen in titanium(Mary's ears perk up!) and our detection ability is so poor that we dont care to add the carbon layer.
Most EM grade silver epoxies seem to have a particle size of 10um, we are trying for something smaller. Master Bond sells a standard grade at 7um and a special grade at 3.5um also the epoxy is a NASA grade for low outgassing. I'm going to try some of the standard grade first and see how it performs.
Ill keep the list updated, if anyone else has other suggestions I'm all ears(well, not ALL).
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.)
Suggest you talk to Ted Dixon, now with BPI in Waterloo, Ontario: 519-886-9013 aedixon-at-confocal.com Ted has worked for years on a confocal macro/microscope which images fingerprints on incredible surfaces... even the black plastic bags which, as he puts it, "seems to be the favorite wrap for murderers and criminals". He also does great physics and should be able to take you through the math to convert from the image to the measurement.
Let me know how it goes.
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 02:06 AM 9/8/99 -0500, Giles Sanders wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You should keep records, blocks, etc. on clinical material at least 10 years. So far, we have never thrown any out, but some is stored off campus. If you want help with CAP go to:
www.cap.org
On Fri, 10 Sep 1999 drennie-at-UNMC.EDU-at-sparc5.microscopy.com wrote:
} Date: Fri, 10 Sep 1999 13:34:37 -0500 } From: drennie-at-UNMC.EDU-at-sparc5.microscopy.com } To: microscopy-at-sparc5.microscopy.com } Subject: Regulations for records keeping in clinical work } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good afternoon, } } I have a question I have not been able to locate the answer to anywhere. I run a clinical } EM lab here at the University of Nebraska and we have recently become associated with the } College of Anatomic Pathologists (CAP) and I am having trouble locating any set } regulations or even guidelines as to the duration we should retain records, as well as } specimen blocks, thick section slides and EM photos. Does anyone have any info to get me } headed in the right direction? } } Thank you, } } Doug Rennie } Coodinator-EM Facility } University of Nebraska Medical Center } Omaha, Nebraska. } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
The phenomenon won't be that strange at all if you take into consideration the following factors: 1. Fracture may include crack nucleation and expansion; 2. In LN, most polymers are brittle macroscopically, but may be ductile microscopically; 3. Loading effects on fracture mode (speed of loading, magnitude and distribution of stress). In your case, when you bend your sample, I would suspect that some pre-existed defects/cracks work as fracture nuclei and because of stress concentration in those locations (specifically at the sharp tips of the microcracks according to fracture mechanics), cracks expand much more easily and rapidly than the cutting case where the tip of the cut may actually be blunt microscopically depending on how good your Japanese knife is.
cy, PhD Rodel, The King of Polishing 451 Bellevue Rd Newark, DE 19713
On Thu, 9 Sep 1999, Zeng Jijun wrote:
} Date: Thu, 9 Sep 1999 21:02:22 -0500 } From: Zeng Jijun {sentoray-at-izu.co.jp} } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM - sampling method of PET } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Recently I got very interesting results on PET/PMMA extrudate } morphology(10 ~ 1 wt% of PMMA and PET was crystalline). Instrument: } Hitachi S800 SEM Sampling method: hand fracturing or knife cutting under } liquid nitrogen temperature. Results: Unexpectedly, hand fracturing sample } has little matrix burrs while knife cutting sample has many. This is } strange because it is usually think cutting is better way than fracture. } Any suggestions? Thank inadvance for your help. Dr. Zeng JijunVisiting } Scholar of Toray Home Address: Nakatogari 734-2-5 Nagaizumi-cho, } Sunto-gun Shizuoka, 411-0942 Japan } Tel: } +81-559-884601(H)E-mail: sentoray-at-izu.co.jpPersonal } Homepage:http://www.geocities.com/ResearchTriangle/Forum/1786 } } } }
I archieved good labelling after stabilisizing my sections with 2% aqueous Glut. Without Glut, my signal was very weak. But collegues in my lab avoid this step, they say it caughts a lot of dirt or they see no diffence. So I would propose you should try it with Glut if your Immuno shows weak staining (with low background!) and you would like to enhance it.
Best wishes and good luck, Michael
Tom Phillips schrieb: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Many EM immuno protocols using colloidal gold on grid mounted thin } sections employ a 1% glutaraldehyde step after the grid is incubated } in the secondary gold-conjugated antibody and washed. Does anybody } know whether this step is really necessary (i.e., is there a paper } that compares fix vs no fix after labeling). I would have thought } the affinity of the antibody would have precluded the need for this } step. Comments welcome. } } 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)
Hi Folks, Does anyone have a feeling for the electric fields that a heated filiment sees as the "extraction field" in a typical TEM or SEM electron gun? Does this change for the Hexaboride filiments?
What is the effective emitting area of the cathode?
best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
I have two rooms which I plan to convert into a confocal microscope suite during the makeover of our building. Our present experience is that the microscopes are sensitive to low frequency vibration caused by foot traffic in corridors and nearby stairs, closing of doors and movement of lifts. I want to minimize the effects of seismic and acoustic disturbance in the new room as near absolutely as is practicable, and proposed to the architects that we construct a vibration-isolated floor. This sounded like a brilliant idea until I was asked how exactly I would recommend them to do this. I have heard anecdotal accounts of a wide variety of solutions, many of which were outrageously expensive or totally impractical. Can anyone recommend the method that combines the *least* bangs per buck with proven efficacy?
Yours sincerely
Chris Jeffree ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
The emitting area of a thermionic filament is typically significantly smaller for LaB6 than for Tungsten (I believe approximate numbers will be 5-15 microns for LaB6 and around 30 for tungsten hairpin). The area depends sensitively on Wehnelt bias.
Several textbooks have good sections treating this, for example the section In L. Reimer's textbook (Transmission Electron Microscopy, Springer-Verlag). In that text, there is a plot of calculated equipotential lines around a filament, with a reference to an article by Haine and Einstein (not Albert), J. Appl. Phys. vol. 3, (1952) p. 40
Regards, Wharton
------- } From: "Dr. Mark W. Lund" {lundm-at-physc3.byu.edu} } To: microscopy-at-sparc5.microscopy.com } Cc: lundm-at-physc3.byu.edu } Subject: Filiment fields } Date: Sat, Sep 11, 1999, 2:53 AM }
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} Does anyone have a feeling for the electric fields } that a heated filiment sees as the "extraction field" } in a typical TEM or SEM electron gun? Does this change } for the Hexaboride filiments?
Judging from the two assemblies for my JEOL SEM, i.e., a W assy and a LaB6 assy, the answer to your question is that the electrostatic fields are essentially the same, but that there are slight differences. That is, both my wehnelt assemblies are identical unless you get out the calipers and actually measure.
I made a program for you. It's a 32 bit Windows program. You can select as many LINK spectrum files (in their original, not converted format) as you need. All the selected files will be converted in one run. I have = tried it with more than 1700 spectra. The result of the conversion is a text = file with the same name but with different file extension. An example from a converted spectrum file (from a series): -------------------------------LAK11S103.SP------------------------------= ------ lak11s** 3
preset live time: 40 live time: 40 real time: 49 20.000000 eV/channel
Energy[keV], counts -0.200000, 0 -0.180000, 0 -0.160000, 0 -0.140000, 0 -0.120000, 3 -0.100000, 16 -0.080000, 42 -0.060000, 98 .. -------------------------------------------------------------------------= ------------ If you are still interested in a program like this, drop me a line and = I'll send it to you.
With the best regards: Laszlo Varga -----Eredeti =FCzenet----- Felad=F3: Paul Rennie (KIDDE) [SMTP:Paul.Rennie-at-kidde-hq.com] K=FCldve: 1999. augusztus 26. 17:29 C=EDmzett: Microscopy-at-Sparc5.Microscopy.Com T=E1rgy: EDX - Link AN10000 file conversion
------------------------------------------------------------------------ Dear list members,
We currently run a Link (now Oxford Instruments) AN10000 EDX system and = would like to convert all of our archived spectra into a format which can be read by a PC. [...] =20 Has anyone come across, or written a routine to perform this as a batch function?
Does anyone have a clever way of dealing with cryosections that stick to the knife? I can't dislodge them with a paintbrush or probe. The sections are stuck only along the cutting edge. Pulling them off with tweezers sometimes works, but more often than not they tear. The sections won't ribbon (which is okay, I prefer to get them one at a time) they just pile up. I tried touching the roll guard so that the section would stick to it, but that doesn't work. The sections are very nice otherwise.
Stephen J. Beck wrote: ========================================================= I have a former student who is working with hyaluronic acid microspheres. These microspheres are clearly visible under the stereomicroscope, however, the desire is to view them using SEM. The problem is that when they are applied to a stub and allowed to air dry they disrupt in some way leaving a film layer behind. They wish to visualize the intact microspheres. In addition, they might also want to try observing them using a TEM (having worked with embedding and sectioning spheres/bubbles of surfactant from the lungs I realize how difficult this might be). If they could be embedded and sectioned what stain(s) might be employed?
The following is a description of how these microspheres are prepared. Any assistance with even just a SEM protocol would be greatly appreciated!
HA Preparation: " A hyaluronic acid (HA)solution and an organic oil (with a small amount of emulsifying agent) are mixed thoroughly until an emulsion forms. ONce the emulsion forms, polymerizing and cross-linking chemicals are added to cross- link the HA. The mixture is then centrifuged and the oil layer is discarded. The HA microsphere pellet is then washed several times with isopropanol and resuspended in a minimum amount of distilled water and then lyophilized." ================================================ When a system of microspheres is vacuum sensitive, one approach we have taken over the years is to use our own SPI Supplies "Wet Replica" Kit (see our website below for details). It is a silicone based system that cures quickly, creating a "negative" replica of the vacuum sensitive microspheres. When cured, the silicone is then thoroughly rinsed with an appropriate solvent to remove all remains of the microspheres of interest. The microspheres are now represented as hollow spheres in the "negative".
Using another component from the kit, we then replicate the replica generating a "positive" replica from the negative. The positive is very stable, can be gold coated conventionally and examined by SEM.
There is one potential drawback of this method, and that is that above 700X, one starts to see structure from the replicating materials. However, for particle size measurements and structural observations below about 700X, this system should work just fine. We have not ourselves used it on HA, but on similar organic vacuum sensitive materials, including vacuum sensitive catalyst particles, it has worked just fine.
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 ######################## ============================================
} Date: Thu, 09 Sep 1999 09:09:03 +1200 } To: Microscopy-at-MSA.Com } From: Alfred Harris {a.harris-at-waikato.ac.nz} } Subject: SEM of Bacteriophage } } Hi everyone } I have had a request for electron microscope images of bacteriophage. I am familiar with standard TEM negative staining methods. Is it possible to obtain SEM images? Are they as good? What are best methods? } } Alfred Harris
"Fazio-Zanakis, Maria, HMR/US" wrote: } My only concern is when changing the } filaments...maybe some residual x ray exposure. But I don't see any big } problem.
There can't be any x-ray exposure as there is no x-ray output as there is no electron beam hitting anything to cause x-ray emission. This is self-evident (no e-beam) as you are changing the filament..
"Residual radiation" from x-ray tubes etc. with no HV and no filament heating is just an urban myth. It's impossible.
So, don't worry about changing filaments to your EM.
I have been trying to get decent plastic sections (3-5 um) of a succulent halophyte species, Salicornia virginica (perennial pickleweed). Preliminary work was done with paraffin embedding and sectioning. Results were good enough to get basic embryological sequence but really need better and thinner sections, especially for the very early prepollination stages. I have been using NaPO4 buffer pH 7.6, adjusted to match osmolality of tissue ( up to 1000 + msosm). Did inital fixation in 2.5 % glut + 2% paraformaldehyde + drop DMSO in .05 M buffer. Postfixed in 2% osmium ( also in buffer of same osmolality), then rinsed and dehydrated in graded EtOH. Finally embedded in LR White resin. All done on ice. Results are very poor. tissue appears badly shrunken and distorted, much of the cellular contents are gone. I have done the fixation without the DMSO and have also tried just fixing directly in osmium. It seems to me that the dehydration is what is causing the damage. I would appreciate any suggestions from the group. thanks, Mary Pfauth, Portland State University.
We have our own Ca-ratio imaging system with Fura-2. We developed the system in SCIL Image 1.4, a multipurpose image analysis platform. You can buy turnkey systems, but make sure they do it right:
The speed of the excitation filter canger is limiting for the speed of the application (besides equilibration of your Calcium-probe), if you use Fura-2. There are severla types of filterchangers possible.
If you also want morphological information and you want to use a camera, the videorate of a classical PAL camera is limited to 40ms time resolution (a bit faster for NTSC). A frame splitter gives you about 10 ms time resolution if it splits the frame in four. Alternatives (digital camera systems) can give you a better time resolution. Systems which only detect the luminance or faster camera-systems give you a better time resoloution.
You have to be careful to make the calculations right, as there are two pitfalls in the procedure:
1) Correct for the darkcurrent of your camera 2) Correct for the background noise, coming from the preparation.
Make sure that the detection system matches the dynamic range of the probe.
Sincerely yours,
Peter Van Osta, MD
Senior Scientist Medical Image Analysis Biological Imaging Laboratory Life Sciences Department I - 6065 Janssen Research Foundation Turnhoutseweg 30 B-2340 Beerse Belgium Europe
-----Original Message----- } From: "Mortro-at-aol.com"-at-sparc5.microscopy.com [mailto:"Mortro-at-aol.com"-at-sparc5.microscopy.com] Sent: Friday, September 10, 1999 10:56 PM To: microscopy-at-sparc5.microscopy.com
Hello all!
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?
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has anybody positive or negative experience with the digital camera DC200 or DC100 for light microscopy sold by Leica? Before we bought DC200 we tested DC100 with lower resolution and found it useful. With DC200 we bought also a lot of problems. The main problems were that the programm under Win NT was not running properly and the white balance does not work. We always get a brown background. As we are investigating fungal infected plant tissues you can imagin that we cannot discriminate between health and necrotic tissue. .Now, we are waiting for month that Leica will solve the software problems with the white balance. It would be interesting for me, if anybody has got the same experiences with this system
Dr. Anne Heller Arbeitsgruppe Elektronenmikroskopie Institut fuer Botanik (210) Universitaet Hohenheim Garbenstr.30 D-70593 Stuttgart Tel.0049-711-459-2180 Fax 0049-711-459-3355
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Cryo-SEM should work fine for this sample. The freezing will stabilize the structure and careful sublimation should effectively remove enough ice to image the microspheres. You did not indicate the diameter of the microspheres but, providing they are not too small (probably not since they are readily seen with the stereomicroscope), they should be readily visible using cryo-SEM. We have done this with microspheres before with good success. For TEM, you may have the best luck with negative staining...again depending partially on size of the microspheres. The prep may take some experimentation....whether fixing will help stabilize the structure (as with using osmium to fix lipid microscpheres) and which negative stain works best. You may want to look at your sample prior to lyophilizing as this step could affect the size and shape of the microspheres.
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 --------------------------------------
Dear Colleagues,
I have a former student who is working with hyaluronic acid microspheres. These microspheres are clearly visible under the stereomicroscope, however, the desire is to view them using SEM. The problem is that when they are applied to a stub and allowed to air dry they disrupt in some way leaving a film layer behind. They wish to visualize the intact microspheres. In addition, they might also want to try observing them using a TEM (having worked with embedding and sectioning spheres/bubbles of surfactant from the lungs I realize how difficult this might be). If they could be embedded and sectioned what stain(s) might be employed?
The following is a description of how these microspheres are prepared. Any assistance with even just a SEM protocol would be greatly appreciated!
HA Preparation: " A hyaluronic acid (HA)solution and an organic oil (with a small amount of emulsifying agent) are mixed thoroughly until an emulsion forms. ONce the emulsion forms, polymerizing and cross-linking chemicals are added to cross-link the HA. The mixture is then centrifuged and the oil layer is discarded. The HA microsphere pellet is then washed several times with isopropanol and resuspended in a minimum amount of distilled water and then lyophilized."
Regards,
Steve
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
RFC822 header -----------------------------------
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Alfred: I expect that FESEM could produce good images showing phages attached to bacteria. Conventional SEM does not have sufficient resolution. If, however, you need to show some structures within the phages or the shape of their heads, FESEM too is insufficient and negative staining/TEM with a final magnification in excess of 400k is required. Negative staining, to show bacteria and phages together at medium powers are rather difficult. Bacteria are too large for good negatively stained images, but you can be lucky. If no FESEM is available you could try your hand doing some metal shadow casting. 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 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Monday, September 13, 1999 6:40 AM, Alfred Harris [SMTP:a.harris-at-waikato.ac.nz] wrote: } } } Date: Thu, 09 Sep 1999 09:09:03 +1200 } } To: Microscopy-at-MSA.Com } } From: Alfred Harris {a.harris-at-waikato.ac.nz} } } Subject: SEM of Bacteriophage } } } } Hi everyone } } I have had a request for electron microscope images of bacteriophage. I am } familiar with standard TEM negative staining methods. Is it possible to } obtain SEM images? Are they as good? What are best methods? } } } } Alfred Harris }
I am interested in receiving comments on the Soft Imaging System's AnalySIS Software including colorview 12 (LM) and Megaview II (TEM) cameras, specifically for biological applications. Please respond to me directly or to the list for comments of general interest. Commercial responses are welcome. Thanks.
Hello All, The LKB Ultrastainer 2168 in my lab is not functioning properly. The stain chamber is not filling completely and there is not enough stain flow through the tubing as well as water flowing through the rinse and wash cycles. The excellent service engineer for Leica is now working at a new company.
I would appreciate any help from anybody as to how to troubleshoot and fix this problem.
In a clinical EM setting, there is not much time allowed for malfunctioning instruments, especially if you work alone. Thanks, Winnie
At several colloidal gold workshops that I have attended, I was informed that the glut. fixation would also prevent the loss of signal during staining. Apparently, in some cases the rapid change in pH that occurs when moving a grid from a buffer or water rinse, into a uranyl acetate or lead citrate stain can cause the gold conjugates to break away from the antibodies. I must admit that I have never tried to stain without the glut. fixation so I don't know if there is any signal loss or not.
Tim Wakefield ----- / 101 Cary Hall / | \ / Auburn University, AL / --|-- \/ 36849 \ | /\ 334-844-3908 \ | / \ ----- \
I am in dire need of a camera for my Philips PSEM 500, but am starting to realize that this microscope is not exceptionally common. I understand that the same photomonitor module was used on the Philips STEM 400.
Does anyone out there have such a camera for sale?
Please? Pretty Please?
Paul Grover Microvista Laboratory Lafayette, Indiana USA
Additional comment on imaging microspheres which have been prepared by low temperature methods. Make sure you use the very lowest kV 2? if you must 3 and really low beam currents in the 10-20 pA range. If you have any ice in or around the sample, they are very beam sensitive.
Good luck
Patrick Echlin Cambridge UK On 13 Sep 1999, Debby Sherman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Cryo-SEM should work fine for this sample. The freezing will stabilize } the structure and careful sublimation should effectively remove enough ice } to image the microspheres. You did not indicate the diameter of the } microspheres but, providing they are not too small (probably not since they are } readily seen with the stereomicroscope), they should be readily visible } using cryo-SEM. We have done this with microspheres before with good } success. } For TEM, you may have the best luck with negative staining...again } depending partially on size of the microspheres. The prep may take some } experimentation....whether fixing will help stabilize the structure (as with } using osmium to fix lipid microscpheres) and which negative stain works } best. } You may want to look at your sample prior to lyophilizing as this step } could affect the size and shape of the microspheres. } } 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 } -------------------------------------- } Date: 9/10/1999 1:28 PM } } From: Steve Beck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues, } } I have a former student who is working with hyaluronic acid microspheres. } These microspheres are clearly visible under the stereomicroscope, } however, } the desire is to view them using SEM. The problem is that when they are } applied to a stub and allowed to air dry they disrupt in some way leaving } a } film layer behind. They wish to visualize the intact microspheres. In } addition, they might also want to try observing them using a TEM (having } worked with embedding and sectioning spheres/bubbles of surfactant from } the } lungs I realize how difficult this might be). If they could be embedded } and } sectioned what stain(s) might be employed? } } The following is a description of how these microspheres are prepared. } Any } assistance with even just a SEM protocol would be greatly appreciated! } } HA Preparation: } " A hyaluronic acid (HA)solution and an organic oil (with a small amount } of } emulsifying agent) are mixed thoroughly until an emulsion forms. ONce the } emulsion forms, polymerizing and cross-linking chemicals are added to } cross-link the HA. The mixture is then centrifuged and the oil layer is } discarded. The HA microsphere pellet is then washed several times with } isopropanol and resuspended in a minimum amount of distilled water and } then } lyophilized." } } Regards, } } Steve } } } } } Stephen J. Beck } Associate Professor } Bio-Imaging Center/Electron Microscopy } Department of Biology } Nassau Community College } Garden City, NY 11530 } Voice Mail: (516) 572-7829 } Email: {becks-at-sunynassau.edu} } URL: {http://www.sunynassau.edu/webpages/biology/becks.htm} } } } } } } RFC822 header } ----------------------------------- } } Received: from SPARC5.MICROSCOPY.COM by mailcenter.btny.purdue.edu } with SMTP (QuickMail Pro Server for Mac 2.0r1b2); 10 SEP 99 } 18:47:57 UT } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com } (8.6.11/8.6.11) id NAA15527 for dist-Microscopy; Fri, 10 Sep 1999 13:14:57 -0500 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id NAA15517 for } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 10 Sep 1999 13:14:26 -0500 } Received: from lib.acs.sunynassau.edu (LIB.ACS.SUNYNASSAU.EDU } [198.38.8.2]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id NAA15510 for } {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Sep 1999 13:14:10 -0500 } Received: from nov1.acs.sunynassau.edu ([198.38.9.253]) } by lib.acs.sunynassau.edu with ESMTP } for Microscopy-at-MSA.Microscopy.Com; Fri, 10 Sep 1999 14:22:14 } -0400 } Received: from NCC_VOL2/SpoolDir by nov1.acs.sunynassau.edu (Mercury } 1.40); } 10 Sep 99 14:28:31 -500 } Received: from SpoolDir by NCC_VOL2 (Mercury 1.31); 10 Sep 99 14:28:29 } -500 } Received: from [198.38.8.22] by nov1.acs.sunynassau.edu (Mercury 1.31) } with ESMTP; } 10 Sep 99 14:28:24 -500 } X-Sender: becks-at-nov1.acs.sunynassau.edu } Message-Id: {l03010d03b3fefb765ba6-at-[198.38.8.22]} } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } Date: Fri, 10 Sep 1999 14:28:42 -0400 } To: Microscopy-at-Sparc5.Microscopy.Com } From: Steve Beck {becks-at-sunynassau.edu} } Subject: Microsphere Preparation } Errors-to: Microscopy-request-at-sparc5.microscopy.com } } } } }
Patrick Echlin Cambridge UKOn Mon, 13 Sep 1999, jean michel Wulveryck wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear microscopists, } I'm looking for the thermal conductivity of kraft. could someone give me } this feature. } Thanking You in advance, } Jean-Michel } } }
I am looking for a 3rd party to provide service and repairs to a used sem we will be purchasing in Nov. The prime SEM candidates are JEOL 6400FE or Hitachi 4000FE.
Bill Delaney Senior Fab Engineer Digital Optics Corp. Charlotte, NC bill-at-doc.com {mailto:bill-at-doc.com}
Jennifer Taylor wrote: ====================================== Recently I have embedded biological polymer fibers in epoxy to section for TEM samples. Unfortunately, the fibers with very small diameters dissolved in the epoxy. The fibers are used for applications requiring the polymer to be absorbed in the human body. Therefore, they also dissolve in water. Does anyone have any suggestions for either alternate methods to obtain a TEM sample or ways to alleviate this problem? ======================================= We have had this kind of problem with small polymer particles which tend to dissolve in any of the standard embedding systems. We solved this problem by taking the following approach:
• Take a flat clear embedding mold, for example, like our SPI 2442C-AB (see website address given below) and fill it half way with either SPI Pon™ 812 or one of the other popular "Epon® substitute" resins.
• After polymerizing into a block, apply sparingly some of the fibers, preferably in a dry form. It deposited from a liquid, surface tension forces interfere with being able to coat the underside of the fibers.
• Then metallize with Pt in a sputter coater. Au is OK if you don't have Pt, but Pt tends to be less likely to smear when thin sectioning.
• Then "disturb" (by shaking, or a slight blast with a duster) the fibers to the extent that at least some have been moved with the unmetallized side up and metallize again. We do this three times. The idea is to encapsulate the fibers in a passivation layer to provide protection from contact with the embedding resin.
• The cavity can then be filled up with resin and when cured, the block sectioned, resulting in undisturbed and umodified (by the resin) cross- sections for TEM. The resin should be put in in two steps, the first one being a very thin additional layer with the second layer deep enough to fill the cavity.
We would be very interested in doing a demo run for you using osmium instead of platinum since in theory at least, an osmium coating should be better than platinum (because of its amorphous nature and zero grain size). Let us know if you would be interested in our doing this.
Disclaimer: SPI Supplies manufactures and distributes some of the products mentioned so we would have a vested interest in their greater use. We have also been performing these kinds of laboratory analytical services since 1970.
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 ######################## ============================================
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all,
I notice lots of calls for recommendations of software, CCD cameras, = diamond knives and other microscopy-related equipment appearing on this = list.
As I too am interested in comments about these products, and as I know = most replies occur off-line where I don't get to see them, wouldn't it be = great if there was a central site where we could submit our own reviews of = the equipment we use, and read other users comments?
At one of the on-line book sellers, the site allows for submission of book = reviews. When added together, the sum of these reviews gives a new reader = a good idea of what to expect. =
Couldn't we do this for our stuff too? In addition to giving the buyer an = informed view of expensive equipment, it might help suppliers identify = potential problems early enough to correct them.
Any comments, volunteers etc.?
Best regards,
Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 213 273 8026 http://www.hei.org/htm/aemi.htm
by williams.edu (PMDF V5.2-32 #39697) with SMTP id {0FI000GJZY90CZ-at-williams.edu} for microscopy-at-sparc5.microscopy.com; Mon, 13 Sep 1999 20:48:36 -0400 (EDT) Received: from localhost by colrain.williams.edu (5.65v4.0/1.1.8.2/16Jul96-0543PM) id AA13780; Mon, 13 Sep 1999 20:48:35 -0400
I've followed previous threads on printers, read published reviews, and spoken with the sales rep. Now I am seeking the opinions from users on the output, reliability, and service of four full-page, color, 300 dpi (or better) printers:
- Sony UP-D70A - Kodak 8660 or 8670PS - Fuji Pictography 3000
- also Codonics printers (sorry, no specific model)
We've been very pleased with the Epson 740 and 750 inkjet printers and glossy film, and now want to improve image quality and printing speed. We're running PCs on a LAN and can connect via parallel port, USB, Adaptec SCSI interface, or Ethernet. Our principal output will be high-resolution images embedded in desktop publishing software such as Word (and some Postscript programs too).
I am seeking opinions on a microspectrophotometer from SensorPhysics and Ocean Optics, called the Lambdascope. If you've used this system and are willing to share your opinions, please contact me off-list.
Hi all, A colleague has asked me to find out if anyone knows of any papers published on the use of environmental SEM to look at slurry. Any references or other information would be appreciated. Thanks.
Lyn Waterhouse CEMMSA Centre for Electron Microscopy and Microstructure Analysis University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356
Thanks to all the people who sent in suggestions; once again, this list has saved the day (or more accurately, the week).
Linda Barthel's ( http://www-personal.umich.edu/~praymond/protocol.html ) suggestion of cutting the block face into a diamond shape did the trick. And no compression at all !!! And you can forgo the roll guard if you're careful (though it worked for me with the roll guard as well).
In accumulating what I want I have come up with some extra Nikon, Zeiss, AO and Swift stuff that I would like to trade for things more useful to me. Everything is subject to return if is not what it is represented to be. I ask the same for what I trade for. I am looking for functional thing and cosmetics are not a sticking point. Optical quality is.
What I have:
A Zeiss Epi attachment for material work with an epiplan 40/.85 objective. It has bright field and dark field. It has a couple of filter slots and a diaphragm. It looks to be in great shape. This is the light tube that attaches to a stand and includes the turret. There is also an Attachment that allows it to attach directly to a binocular head.
A Zeiss condenser that probably went with the epi stuff it has a Epiplan .63 and 0.32 lens with two swing out filter trays that snap into place. The condenser has centering screws.
A focusing gear for the cream colored Zeiss. I wish I had got to it before some scraped it for metal:{
A black Nikon triocular head with a 1.2 relay lens and lighted pointer. The camera port is odd in that it comes off horizontally instead of vertically. It is for a TV camera and is not of standard eyepiece size. It has some dings in the paint but nothing that can't be detailed out. It says CirCon MV9585 micro optical system.
I have the Nikon binocular eyepiece head for it. It is a 95108 and has some chipping on the paint and needs cleaning. It has been forced from the retting ring and has a couple of ding on the curricular dove tail.These are not bad and can easily be filed out with no loss of functionality.
A Zeiss binocular eyepiece head is cream and folds in the center. It needs cleaning and the paint needs retouching here and there. There no dings or dents. Looking in the top at a good angle I can see some dirt or discoloration on the edges of one of the prisms where it is attached to the scope. This cannot be seen looking directly into the eyepieces and with eyepieces in the head. It appears to be normal to the mounting process.
A AO cycloptic that has a loose prism. It shows use. No stand or eyepieces. I would like to get the prism repaired and find a stand for this.
A 170 mm Leitz missing eyepieces and condenser but having a triouclar head. This is the 50 or 60 model that the course and fine focus is on the same knob. I really intend to keep this scope and most of what I am looking for is for it.
A Binkmann Medical scope that I believe was made by Zeiss in east Germany. Binkmann swears it is Zeiss and Zeiss says They never made it. It is a nice scope to use and is very nice condition. I am very happy with the scope. The only reason I mention it is that it might be of interest to a Zeiss collector if it is in fact made by Zeiss.
A 1 and 2 x head for a Swift binocular. It uses large Eyepieces and is missing eye pieces and stand. I would either like to find eyepieces and a stand or trade this. If I can find a stand I will machine adapters for standard eye pieces.
I have a Zeiss IKon 35mm w/focal plane shutter and front reflex housing and an attachment to eyepiece tube with photo eyepiece. I also have a single eyepiece tube. I know where there is a Second I can lay hands on if the deal is right. I are not very interested in trading these but would do it for the right stuff. Is in excellent shape
Now my want list. I am willing to trade most anything to end up with what I want.
I need some eyepieces; a pair of 8x, 15 x and a couple pairs of 10 x.
A good condenser for the Leitz. It is the one that rides on a Dovetail.
I would like a good phase setup for the Leitz and darkfeild would Be nice.
I need a stage micrometer. And I would be open to trade for some books on invertebrates and protozoa.
I need a stand for the Cycoplotic if I can get it fixed or I need a Stand for the Swift.
I would like to have a triocular head for the Binkman. It has A larger flange than the Zeiss. I can turn an adapter ring if necessary.
I am interested in adding functionality to the 170mm Leitz
A couple of mirrors.
A good light source.
I am currently in the San Francisco area for the next couple of weeks. I have some of the stuff with me. wold prefer to do business in person but I have done a great deal of internet business and have no problems with that.
My goal is to end up with two working binoculars and two working compounds with TV capability. Eyepiece cameras will be satisfactory For the binoculars. There are two of us working on this and most of My work under 100x will be done with a macro camera not a binocular Microscope. We have the video stuff already.
I am a ham radio operator and I have a good collection of stuff and access To a lot more. So if you want something in the electronics line I might have it.
Gordon Couger
408 249 7483 until the 27 of September. Call afternoon or evenings. Gordon Couger 624 Cheyenne Stillwater, OK 74075 405 624 2855 GMT - 6:00
I will setup Q & A forum about equipment at my Microscopy Vendors Database (http://www.kaker.com/mvd/vendors.html).
Henrik
Paul Webster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear all, } } I notice lots of calls for recommendations of software, CCD cameras, diamond knives and other microscopy-related equipment appearing on this list. } } As I too am interested in comments about these products, and as I know most replies occur off-line where I don't get to see them, wouldn't it be great if there was a central site where we could submit our own reviews of the equipment we use, and read other users comments? } } At one of the on-line book sellers, the site allows for submission of book reviews. When added together, the sum of these reviews gives a new reader a good idea of what to expect. } Couldn't we do this for our stuff too? In addition to giving the buyer an informed view of expensive equipment, it might help suppliers identify potential problems early enough to correct them. } } Any comments, volunteers etc.? } } Best regards, } } Paul Webster, Ph.D. } House Ear Institute } 2100 West Third Street } Los Angeles CA 90057 } 213 273 8026 } http://www.hei.org/htm/aemi.htm
} Chesapeake Society for Microscopy } presents } The Fall Dinner Meeting of 1999 } } Date: October 6th, 1999 } } Time: Social Hour at 6:00 PM } Graciously Sponsored by Ray Gundersdorff, JEOL } } Dinner: 7:00 PM--$20.00 (Students $10.00) } Menu includes several Chinese dishes } } Location: Far East Restaurant } } Speaker: Dennis Ward from the FBI } "Applications of Scanning Electron Microscopy at the FBI" } } } } Far East Restaurant } 5055 Nicholson Lane } Rockville, MD 20852 } (301) 881-5552 } From Beltway, North on 355 (Rockville Pike) } Make Right onto Nicholson Lane (Street right after Whiteflint Mall) } At 3rd light make left into parking lot } } Please make reservations by Oct. 5th } Contact Andrea Weisberg at (301) 435-1977 } Andrea S. Weisberg } NIH/NIAID/LVD } Bldg.4/Rm.210 } 4 Center Dr. } Bethesda,MD 20892-0445 } office (301) 435-1977 } Fax (301) 480-1147 } e-mail: aweisberg-at-nih.gov } }
ATHENE DONALD (Cambridge University) is one of our UK experts on ESEM - perhaps these three articles might be helpful:
(2) TI: The study of water in heterogeneous media using environmental scanning electron microscopy AU: Thiel_BL, Donald_AM JN: JOURNAL OF MOLECULAR LIQUIDS, 1999, Vol.80, No.2-3, pp.207-230
(14) TI: Direct observation of water-oil emulsion systems in the liquid state by environmental scanning electron microscopy AU: Stokes_DJ, Thiel_BL, Donald_AM JN: LANGMUIR, 1998, Vol.14, No.16, pp.4402-4408
(16) TI: Environmental scanning electron microscopy for the study of 'wet' systems AU: Donald_AM JN: CURRENT OPINION IN COLLOID & INTERFACE SCIENCE, 1998, Vol.3, No.2, pp.143-147
On Tue, 14 Sep 1999, Lyn Waterhouse wrote:
} A colleague has asked me to find out if anyone knows of any papers } published on the use of environmental SEM to look at slurry. Any } references or other information would be appreciated. Thanks.
} Lyn Waterhouse
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
First of all the application framework I will talk about is NOT a commercial application, it is our own in-house developped digital image analysis framework for analysing microscopical images. I want to know if there are any commercial equivalents on the market to what we have here ?
We have developed (already a copule of years ago) a multi-mode multi-position automated image analysis system for micrscopy. The system is based on SCIL Image 1.x (http://carol.wins.uva.nl/~koelma/isis/projects/scilimage.html), an image analysis framwork from the University of Amsterdam in the Netherlands.
In one setup for fluorescence micrscopy, we are capable of acquiring 28380 grey-scale image in less than 4 hours from the central 60 wells of a 90 well plate (473 images per well), including focussing every 4 positions in the well. We use a ZEISS Axiovert 135, with a 40x objective, a motorised stage and an intensified camera to acquire the images. Our (patented) focusing algorithm is capable to focus even if the S/N ratio is 5 dB, ie. low quality fluorescence images.
The position recording system allows us to scan the plates in several modes, ie. different fluorochromes or different imaging modes (brightfield, phase contrast, DIC,...).
Analysis of the 28380 images takes about 9 hours, depending a bit on the application. The acquisition (SGI O2) and the analysis (SGI Origin200) is done on Silicon Graphics workstations.
Are there any commercial systems that are capable of doing this ? Our application is NOT for sale, so this is not a commercail spam.
Sincerely yours,
Peter Van Osta, MD
Senior Scientist Medical Image Analysis Biological Imaging Laboratory Life Sciences Department I - 6065 Janssen Research Foundation Turnhoutseweg 30 B-2340 Beerse Belgium Europe
Hey, a "Comsumer Reports" of microscopy equipment. Great idea. Are you volunteering to catalog them???? Maybe it's a project MSA would support with a small grant to keep the data base????
On 13 Sep 1999, Paul Webster wrote:
} Date: 13 Sep 99 17:44:04 -0700 } From: Paul Webster {pwebster-at-mailhouse.hei.org} } To: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } Subject: Recommnedations } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } } I notice lots of calls for recommendations of software, CCD cameras, diamond knives and other microscopy-related equipment appearing on this list. } } As I too am interested in comments about these products, and as I know most replies occur off-line where I don't get to see them, wouldn't it be great if there was a central site where we could submit our own reviews of the equipment we use, and read other users comments? } } At one of the on-line book sellers, the site allows for submission of book reviews. When added together, the sum of these reviews gives a new reader a good idea of what to expect. } Couldn't we do this for our stuff too? In addition to giving the buyer an informed view of expensive equipment, it might help suppliers identify potential problems early enough to correct them. } } Any comments, volunteers etc.? } } Best regards, } } Paul Webster, Ph.D. } House Ear Institute } 2100 West Third Street } Los Angeles CA 90057 } 213 273 8026 } http://www.hei.org/htm/aemi.htm } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
What you have provided is the URL for Scion Image which was developed from NIH image. That is good to know, too.
However, Image Tool is a different product from the University of Texas Health Sciences Center. It is true that the links below do not work. Unfortunately, they are the ones listed officially on all of the UTHSCA pages that I found. I finally called Dr. Dove, who was involved in developing the program, and he directed me to the correct site. It is available through the main UTHSCA page following a "Resources" link, and finding the "Image Tool" listing. The end URL is http://macorb.uthscsa.edu/dig/itdesc.html
The demand was outstripping their old server and they had to move the site. Also, there is apparently more demand for improving and updating the program itself than there is for updating the web pages.
At 06:50 PM 9/3/1999 -0700, you wrote: } } I think UTHSCA ImageTool was taken over by Scion Image(???) At any rate, } Scion has an easy to use image analysis program that can be downloaded for } free from their site: } } http://scioncorp.com } } Sometimes I have LUT problems, but not when I'm doing image analysis. } (Before I got Photoshop, I used to use it to draw pictures ;-) ) } } ------------------------------------- } } On Fri, 3 Sep 1999, Andrew Ochalski wrote: } } } } } Dear Microscopists, } } } } I have been trying in vain for the past two } } weeks to download the most recent version of } } UTHSCSA's ImageTool program. The two links } } I have been trying from a variety of approaches } } are: } } http://ddsdx.uthscsa.edu } } and } } ftp://maxrad6.uthscsa.edu. } } Are these still valid, are there more current links } } or is the software no longer available ? Thanks } .. }
Hi all - taking a slurry as any mixture of solid particles in liquid? we have looked at a few fibre/oil mixes, and found the influence of kV to be crucial. 25kV imaged particles through the surface of the liquid without giving any detail of the liquid, at 10kV the liquid/gas boundary was more apparent, but at 5kV there was quite a dramatic shift, as detail on the surface of the liquid phase became visible (mixed SE/BSE signal) - what looked like a perfectly smooth surface even at 10-15kV was actually covered in gunk or had its own microstructure. You need (IMHO) helium (around 1 torr. give or take a bit) in the chamber to work comfortably at 5kV. Lower kV than 5 is probably practical on the newer model VP or ESEMS.
But if its a pure water slurry you are looking at, maybe it would be possible to drop the temperature as much as you can, and to mix in something else, to cut the evaporation rate to let you work at lower pressure to let you work at lower kV....?
cheers
Sally
Dr Sally Stowe Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475, ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525 http://www.anu.edu.au/EMU/home.htm
} } } Lyn Waterhouse wrote:
Hi all, A colleague has asked me to find out if anyone knows of any papers published on the use of environmental SEM to look at slurry. Any references or other information would be appreciated. Thanks.
Lyn Waterhouse CEMMSA Centre for Electron Microscopy and Microstructure Analysis University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356
I use a Kodak DS8650PS dye sublimation printer with Kodak ExtraLife XL paper and ribbon, 8-1/2"x12" paper. It is connected to a 10BaseT LAN with Macs and PCs (Win95) and HP Laserjets.
The quality of the Kodak is very good. However, each page costs about $2, between the cost of the paper and the ribbon. Printing speed is rather slow, even with 64MB of RAM. Sending anything higher than 300dpi is a waste. 220 dpi works well and does speed things up. The printer is a CMYK + coating so it makes 5 passes. Each print takes about 3-4 minutes to print. But it can take up to 5 minutes to just get the data file into the printer. So from hitting the Print command in a program, you can expect to wait at least 10 minutes for a printed page to emerge.
The ExtraLife is very good stuff since it is for all practical purposes, indestructible and does not fade with age. You can soak it in water and just wipe off and there is no damage to the print.
I have had sporadic luck doing direct printer port outputs to the 8650PS. But the Ethernet link never fails from the PC or Mac.
These printers can be found used for between $2,000 and $4,000. The newer models are supposedly faster but I'd suspect the difference is not all that much. But I have not seen one in action to verify that.
gary g.
At 05:48 PM 9/13/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have the space to keep the data base. I don't have the expertise to edit it. It is a pretty good link. It is not as fast as the best but the congestion is low enough it makes up for it most of the time.
I will consider any form of internet publication. I am looking for a small circulation juried journal. I am primarily interested in increasing the speed of information distribution and reducing the cost.
My experience with some of my own publications where it took a year to get it published after acceptance and $700 bucks for reprints and they kept the copyright left a bad taste in my mouth.
If we are going to increase the rate of progress in technology we need to get the time to publication down as low as possible. I think it should be possible to get a non controversial jurried article out in two months and letters out in hours.
I am interested in publishing only. I am no editor. All copyrights stay with the author and no page cost at this point. The cost of publication on the internet is very low. My cost for keeping a box on the net are 50 a month and hardware replacement. $1,000 a year covers the cost including administration after the first time set up. So very modest page charges would be very attractive to and ISP. My interest is getting it started.
My cost will go up with a lot of net acess. But not much.
Gordon Gordon Couger 624 Cheyenne Stillwater, OK 74075 405 624 2855 GMT - 6:00 -----Original Message----- } From: Sara Miller {saram-at-duke.edu} To: Paul Webster {pwebster-at-mailhouse.hei.org} Cc: MSA listserver submission {Microscopy-at-Sparc5.Microscopy.Com}
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We are considering revamping our billing software in a multi-user, multi-instrument imaging facility. Can anyone suggest a good software package that they currently use for billing?
Thanks in advance for any suggestions.
Doug Taatjes
Dr. Douglas J. Taatjes Department of Pathology Director, Cell Imaging Facility University of Vermont Burlington, VT 05405 USA 802-656-0373 (voice) 802-656-8892 (FAX)
Web Page: http://pathology.uvm.edu/cifweb/cif_home/cif_index.html
Glutaraldehyde fixation after immunogold staining prevents the loss of signal that may be induced by low pH contrasting, such as in Uranyl salt solutions. Next to this Uranyl salts may have chaotropic effects. Both chemical characteristics are exploited in eluting antibodies form antigens in affinity chromatography purification techniques, as well as in eluting immunoglobulins from Protein A or Protein G affinity columns. In our experience the effect of the fixation step is more obvious with protein A or G reagents than with secondary antibody gold conjugates, where it will depend on Kd-values for the individual reactions beteen primary and secondary antibody. In fact Protein A gold may even become uncoupled from the primary antibody simply by washing in water. Of course it is necessary to rinse well after the gold incubation step before the on-set of fixation, since all adhering gold conjugates may become fixed to the specimen, both the ones bound to the primary as the unbound ones which should first be removed by washing. The fixation time can be relatively short, in the order of minutes, for on-section labeling and may have to be applied for a longer period of time for pre-embedding applications.
Hope this helps to clarify this =========================== Jan Leunissen AURION http://www.aurion.nl Costerweg 5 6702 AA Wageningen The Netherlands phone: 31-317-497676 fax: 31-317-415955 You will find more technical info on our web site
I notice lots of calls for recommendations of software, CCD cameras, diamond knives and other microscopy-related equipment appearing on this list.
As I too am interested in comments about these products, and as I know most replies occur off-line where I don't get to see them, wouldn't it be great if there was a central site where we could submit our own reviews of the equipment we use, and read other users comments?
At one of the on-line book sellers, the site allows for submission of book reviews. When added together, the sum of these reviews gives a new reader a good idea of what to expect. Couldn't we do this for our stuff too? In addition to giving the buyer an informed view of expensive equipment, it might help suppliers identify potential problems early enough to correct them.
A super idea! The problem for me as for anyone else who volunteers will be finding the time but it will be well spent. Count me as a volunteer! Rosemary
Our Fuji Pictrography has developed a problem where the donor paper gets jammed almost every cycle (often at 1/2 but also at further along the pathway). Have any Fuji owners had this problem and solved it? Any help would be gratefully appreciated.
Someone asked recently about high end printers including the Fuji. We love ours but the St. Louis Service rep that is responsible for our territory is terrible and is hard to even get to them to return a call. Fuji doesn't have a help line, they refer us to our rep. So despite having a great printer, their technical help suffers.
Thanks, Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I totally agree with this, Journal review is taking far too long now. Scientists should really put more effort in reviewing their co-workers paper submissions. Papers sitting on someone's desk for six months without review is unacceptable. I personally try to finish a review once I receive it within two weeks. Many people asked to review papers often never reply to the publishers, which I think is a serious offence in the scientific field.
I know the NIH is considering doing rapid article publication on their website. After screening of the articles, they are placed onto the web for anyone to download or read. I think there are other journals going this route as well. It is the communication method of the future. I think that publications will go this way. More respected online journals will have extensive peer review and screening. Other rapid publication journals will have less, and still be immensely useful in being able to do online searching for specific information.
I have a lot of experience in programming, web design, and system administration. I am interested in forming a rapid publication web site for the scientific community and will readily offer my skills and time. Gordon Vrdoljak.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak 1 Cyclotron Road ICQ 23243541 http://nature.berkeley.edu/~gvrdolja MS90-1116 GAVrdoljak-at-lbl.gov Ernest Orlando phone (510) 495-2829 Lawrence Berkeley fax (510) 486-7797 National Laboratory cell (510) 290-6793 Berkeley CA 94720
On Tue, 14 Sep 1999, Gordon Couger wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have the space to keep the data base. I don't have the expertise } to edit it. It is a pretty good link. It is not as fast as the best but } the congestion is low enough it makes up for it most of the time. } } I will consider any form of internet publication. I am looking for } a small circulation juried journal. I am primarily interested in } increasing the speed of information distribution and reducing } the cost. } } My experience with some of my own publications where it took } a year to get it published after acceptance and $700 bucks for } reprints and they kept the copyright left a bad taste in my mouth. } } If we are going to increase the rate of progress in technology we } need to get the time to publication down as low as possible. I think } it should be possible to get a non controversial jurried article out } in two months and letters out in hours. } } I am interested in publishing only. I am no editor. All copyrights } stay with the author and no page cost at this point. The cost of } publication on the internet is very low. My cost } for keeping a box on the net are 50 a month and hardware } replacement. $1,000 a year covers the cost including administration } after the first time set up. So very modest page charges would } be very attractive to and ISP. My interest is getting it started. } } My cost will go up with a lot of net acess. But not much. } } Gordon } Gordon Couger } 624 Cheyenne } Stillwater, OK 74075 } 405 624 2855 GMT - 6:00 } -----Original Message----- } } From: Sara Miller {saram-at-duke.edu} } To: Paul Webster {pwebster-at-mailhouse.hei.org} } Cc: MSA listserver submission {Microscopy-at-Sparc5.Microscopy.Com} } Date: Tuesday, September 14, 1999 4:22 PM } Subject: Re: Recommnedations } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hey, a "Comsumer Reports" of microscopy equipment. Great idea. Are you } } volunteering to catalog them???? Maybe it's a project MSA would support } } with a small grant to keep the data base???? } } } } } } On 13 Sep 1999, Paul Webster wrote: } } } } } Date: 13 Sep 99 17:44:04 -0700 } } } From: Paul Webster {pwebster-at-mailhouse.hei.org} } } } To: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } } } Subject: Recommnedations } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Dear all, } } } } } } I notice lots of calls for recommendations of software, CCD cameras, } diamond knives and other microscopy-related equipment appearing on this } list. } } } } } } As I too am interested in comments about these products, and as I know } most replies occur off-line where I don't get to see them, wouldn't it be } great if there was a central site where we could submit our own reviews of } the equipment we use, and read other users comments? } } } } } } At one of the on-line book sellers, the site allows for submission of } book reviews. When added together, the sum of these reviews gives a new } reader a good idea of what to expect. } } } Couldn't we do this for our stuff too? In addition to giving the buyer } an informed view of expensive equipment, it might help suppliers identify } potential problems early enough to correct them. } } } } } } Any comments, volunteers etc.? } } } } } } Best regards, } } } } } } Paul Webster, Ph.D. } } } House Ear Institute } } } 2100 West Third Street } } } Los Angeles CA 90057 } } } 213 273 8026 } } } http://www.hei.org/htm/aemi.htm } } } } } } } } } } } } } } } } Sara E. Miller, Ph. D. } } P. O. Box 3020 } } Duke University Medical Center } } Durham, NC 27710 } } Ph: 919 684-3452 } } FAX: 919 684-8735 } } } } } }
There is an immediate opening for a Manager of a Materials Characterization Facility at the University of Pennsylvania, Philadelphia, PA. The facility is housed within the buildings of the Laboratory for Research on the Structure of Matter (LRSM) and is closely affiliated with the Department of Materials Science. The facility houses primary research equipment for electron microscopy and spectroscopy, ion scattering and scanned probe microscopy. Included in the equipment are a loaded FEG-TEM (JEOL 2010F), HREM (JEOL 4000), FEG-SEM (JEOL 6300F), thermionic SEM (JEOL 6400), STEM-TEM (Philips 400, likely upgraded soon), Scanning Auger (Phi) w/ SIMS, ion accelerator (NEC) w/ three beam lines and two AFMs (Digital).
The University of Pennsylvania is located in Philadelphia, one of the nation's most vibrant cities. The university, a member of the Ivy League, was founded by Ben Franklin and is the fourth oldest, and first secular, university in the US. The LRSM was constructed to house one of the original three Materials Research Laboratories (MRLs) in the US. The university has a continuing tradition of leading materials research and has housed the MRL (now MRSEC) continuously for 40 years.
The text of the official job listing from the University of Pennsylvania website follows. Please refer to the Penn Human Resources website for the official hiring policy of the university at www.hr.upenn.edu. For information about the specific position, please contact Professor David E. Luzzi at luzzi-at-lrsm.upenn.edu.
Text of website listing
Reference Number: 99083647DL Job Title: MANAGER D School/Center: ENGINEERING & APPLIED SCIENCE Department: MATERIALS CHARACTERIZATION Date Posted: 8/30/99 Salary Grade: 028 Minimum: $41,500.00 Top of First Third: $52,533.00 Top of Second Third: $63,566.00 Maximum: $74,600.00 Position Length: Ongoing
Duties: Manage Materials Characterization Facility/Service Center; develop short & long term use agreements with industrial & academic users; assist in experiments by users; compose & present reports bi-weekly to faculty oversight committee; coordinate work assignments of technical staff; assist or coordinate assistance of faculty in teaching & equipment acquisition; train or coordinate training of users; maintain or coordinate maintenance of equipment.
Qualifications: BA/BS in Physical Science or Engineering is required, advanced degree preferred; extensive experience in operation & use of transmission & scanning electron microscopes for imaging, diffraction & spectroscopy; knowledge of vacuum systems, ion scattering techniques, spectroscopic techniques & surface force techniques required.
My thanks to all who replied to my post. Thanks for all the good } suggestions, some of which were suggested by more than one person. My } question for Geoff Williams is this: how does one measure the osmolarity } of cytoplasm only or vacuolar contents only? I have just been grinding tissue, spinning it, and measuring osmolarity of the supernatent. Also, } these plants sequester NaCl in their vacuoles as part of their salt } tolerance mechanism. Don^Ot they require a balancing osmoticum in the } cytoplasm? I thought that was what prolines and betaines did. Do you have any references on this point? Thanks again. Mary
Thanks to all who offered suggestions. Chip Montrose came up with a number for a Fuji Tech line. We have called in the past and they have simply referred us to our worthless dealer. This time we explained our dealer wasn't helping so they connected us with a tech rep. The rep said we could open an account (free for telephone help) and be able to call directly in the future.
The rep then instantly diagnosed the problem as a sticky trip switch adjacent to the cutter for the donor paper. We confirmed this switch was sticking and have a new one on order. Thanks again (esp to Chip!). 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)
At 05:20 PM 9/14/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
[snip]
Another option is to host material and links to materials. Rather than taking on the work of absorbing and managing all material, one would be greatly relieved by only having to manage links. Most folks have web sites available to them. They can post their own materials at those sites and submit those specific URLs. Periodically, a snake can validate each URL for currency. For those folks who do not have web sites available, then of course they would submit the material.
I suspect that hosting only a mass of material could chew up a good sized chunk of disk space. Furthermore, unless the disk is regularly backed up, a crash means that all material will have to either be re-submitted or reconstructed from the last backup.
At 01:39 PM 9/12/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I asked this same question about a month ago. The resounding answer is yes, a SEM will image bacteria. But it is not easy. I'm working on some samples right now and can get good images at about 2,000X but above about 5,000X, the resolution falls off. I'm using a LaB6 instrument and it should have good resolution up to perhaps 70,000X. I think and hope that operator error is the main culprit right now. I'd like to fire the SEM operator but I'm the operator. So, the struggle goes on. But I think that I will succeed.
I would prefer links but I can store the data and the system is regulary backed up. It host some small international web pages and mailer. It is backed up weekly to a second on board hard drive downloaded to CDROM. The last disaster recovery took 45 minutes. A rebuild from CDROM would take a couple of hours right now. This will increase with more data.
With 10 gig drives at $300 and falling the cost of storage does not worry me much.
Gordon Gordon Couger 624 Cheyenne Stillwater, OK 74075 405 624 2855 GMT - 6:00 } } Another option is to host material and links to materials. Rather than taking } on the work of absorbing and managing all material, one would be greatly } relieved by only having to manage links. Most folks have web sites } available to them. They can post their own materials at those sites and } submit those specific URLs. Periodically, a snake can validate each } URL for currency. For those folks who do not have web sites available, } then of course they would submit the material. } } I suspect that hosting only a mass of material could chew up a good sized } chunk of disk space. Furthermore, unless the disk is regularly backed up, } a crash means that all material will have to either be re-submitted or } reconstructed from the last backup. } } gary g. }
I have got six or seven replies for my question on the sputter coater target. Most of them suggested to use a solvent or detergent type liquid like an alcohol, or an acetone. One of them suggested polishing, and another of them suggested to use an acid cleaning. I tried ultrasonic cleaning with acetone and got a reasonably good, even not the best, result. Still some red stuff was left, but the coater works OK.
To the people who responded to my question about using ESEM to look at slurry, thank you for your suggestions - I've taken note of your ideas and appreciate the help.
Lyn Waterhouse CEMMSA Centre for Electron Microscopy and Microstructure Analysis University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356
Chaps, Some time ago there was commercially produced a wall chart (Germany??) that displayed the different types of fracture surfaces (+ a few errors) as observed using an SEM and which was used as an aid / display by metallurgists. Does anyone out there have a similar wall chart in their SEM / Metallurgy lab or perhaps know any of the details of similar types of wall charts. Possibly, it is no longer available?? Thankyou for your help Barry M UNIT UNSW
Barry: I have the answer to your query. One of these charts is hanging in my lab. Here are the detail: Title: FRACTOGRAPHY IN MATERIALS SCIENCE AND ENGINEERING published by ASM International, The Materials Information Society, Materials Park, OH 44073-0002 USA Tel: 216-338-5151 Fax: 216-338-4634
The chart has been photographed and designed by Mohan Chaudhari, Ph.D.,P.E Columbus Metallurgical Services,Inc. 4348 Reynolds Drive Hillard, OH 43026-1260 USA Tel: 614-529-1311 Fax: 614-529-1818 If you have any difficulty please let me know off line. I know Mohan well.
Vin Berry Analytical Technology, GE Plastics, Washington, WV 26181 Tel: 304-863-7528, fax -7108 GE Dial: 8*572-7528 e.mail: vinod.berry-at-gep.ge.com
-----Original Message----- } From: Barry Searle [mailto:B.Searle-at-unsw.edu.au] Sent: Thursday, September 16, 1999 4:17 AM To: microscopy-at-sparc5.microscopy.com
Chaps, Some time ago there was commercially produced a wall chart (Germany??) that displayed the different types of fracture surfaces (+ a few errors) as observed using an SEM and which was used as an aid / display by metallurgists. Does anyone out there have a similar wall chart in their SEM / Metallurgy lab or perhaps know any of the details of similar types of wall charts. Possibly, it is no longer available?? Thankyou for your help Barry M UNIT UNSW
I am in need of the specimen cassette holder (part no 0303) for an A/S 325 microtome. I have tried contacting the company direct but get no response, so i am asking you all whether any of you have one of these microtomes sitting in a corner somewhere gathering dust and would be prepared to part with the holder. If not does any one know where i may be able to obtain one?
Cheers Phil Mutch
Mr Philip Mutch, School of Biomedical Science, E Floor Medical School, University of Nottingham, Nottingham, NG7 2UH. UK. E-mail Philip.Mutch-at-nottingham.ac.uk
While I agree that traditional publication procedures are slow and cumbersome, they do provide an important service: reviewing. Without review, the quality of the publications will quickly degrade to a point that makes publishing useless. Just check out the physics newsgroups on the net. When I last looked there, there were a number of whackos who were arguing for the weirdest ideas (Earth core is made of strawberry jelly kind of thing). I am afraid, that these people would quickly take over any unreviewed publishing site, and make it completely useless for scientific publications.
Also look at the scientific journals: the ones with the strictest screening are in general respected the most.
I would think, that in order to provide a value to the science community, it is a good idea to go digital on the publications WITHOUT sacrificing the quality of the publications, i.e., without giving up reviewing.
I remember a few years back there was a big uproar in the physics community about some research that was published in the New York Times before it made it into a scientific journal (I forgot, what exactly it was. Cold Fusion??).
Just my thoughts...
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: Gordon Vrololjak[SMTP:GVRDOLJA-at-NATURE.BERKELEY.EDU] } Sent: Wednesday, September 15, 1999 11:50:57 AM } To: Gordon Couger } Cc: Sara Miller; Paul Webster; MSA listserver submission } Subject: Re: Recommnedations } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I totally agree with this, Journal review is taking far too long now. Scientists should really put more effort in reviewing their co-workers paper submissions. Papers sitting on someone's desk for six months without review is unacceptable. I personally try to finish a review once I receive it within two weeks. Many people asked to review papers often never reply to the publishers, which I think is a serious offence in the scientific field.
I know the NIH is considering doing rapid article publication on their website. After screening of the articles, they are placed onto the web for anyone to download or read. I think there are other journals going this route as well. It is the communication method of the future. I think that publications will go this way. More respected online journals will have extensive peer review and screening. Other rapid publication journals will have less, and still be immensely useful in being able to do online searching for specific information.
I have a lot of experience in programming, web design, and system administration. I am interested in forming a rapid publication web site for the scientific community and will readily offer my skills and time. Gordon Vrdoljak.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \/\/\ Gordon Ante Vrdoljak 1 Cyclotron Road ICQ 23243541 http://nature.berkeley.edu/~gvrdolja MS90-1116 GAVrdoljak-at-lbl.gov Ernest Orlando phone (510) 495-2829 Lawrence Berkeley fax (510) 486-7797 National Laboratory cell (510) 290-6793 Berkeley CA 94720
On Tue, 14 Sep 1999, Gordon Couger wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have the space to keep the data base. I don't have the expertise } to edit it. It is a pretty good link. It is not as fast as the best but } the congestion is low enough it makes up for it most of the time. } } I will consider any form of internet publication. I am looking for } a small circulation juried journal. I am primarily interested in } increasing the speed of information distribution and reducing } the cost. } } My experience with some of my own publications where it took } a year to get it published after acceptance and $700 bucks for } reprints and they kept the copyright left a bad taste in my mouth. } } If we are going to increase the rate of progress in technology we } need to get the time to publication down as low as possible. I think } it should be possible to get a non controversial jurried article out } in two months and letters out in hours. } } I am interested in publishing only. I am no editor. All copyrights } stay with the author and no page cost at this point. The cost of } publication on the internet is very low. My cost } for keeping a box on the net are 50 a month and hardware } replacement. $1,000 a year covers the cost including administration } after the first time set up. So very modest page charges would } be very attractive to and ISP. My interest is getting it started. } } My cost will go up with a lot of net acess. But not much. } } Gordon } Gordon Couger } 624 Cheyenne } Stillwater, OK 74075 } 405 624 2855 GMT - 6:00 } -----Original Message----- } } From: Sara Miller {saram-at-duke.edu} } To: Paul Webster {pwebster-at-mailhouse.hei.org} } Cc: MSA listserver submission {Microscopy-at-Sparc5.Microscopy.Com} } Date: Tuesday, September 14, 1999 4:22 PM } Subject: Re: Recommnedations } } } } ----------------------------------------------------------------------- - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------
Can anyone help us with fixing our LKB Utrotome IIIs by faxing circuit diagram? Much appreciated.
Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories (MIL) and Department of Biology University of California at San Diego
address: 1500 Bonner Hall, University of California at San Diego 9500 Gilman Drive, La Jolla, CA 92093-0368 telephone - lab: 8585342484 telephone - office: 8588223373 pager: 8586161420 fax: 8588223715 email: mmm-at-biomail.ucsd.edu www site: http://mil.ucsd.edu
I am seeking your help in preparing a multimeric protein (800kDa) -oligomeric form. I used a single C layer film + negative stain (UA) and would like to know if any one has used a double C layer technique. Rosemary
We will replace our primary SEM with a new ESEM in a few months. Consequently, some of our older instrumentation will become available for excess, surplus, or some other sort of disposition (spare parts?). Please e-mail if there is there any interest for the following items? Note: the items will be disposed by the usual official government protocols.
1. Cambridge S-250 SEM, was working well until CRT problem this past May (horizontal line showing on screen). May be corrected for $5000-$10000 by Leo, Inc. although some parts are not available any longer.
2. Tracor-Northern 2010 EDS, still functional as of May, 1999. Replaced CPU board this past January.
3. Microspec WDX-2A with detector, probably functional. Since Oxford can upgrade these WDS units, we may keep it. Not my decision.
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124-4305
Dear Listers, I'm lookling for a commercial source in the US for a low viscosity vinyl silicone (dental impression material). I found a commercial product in a paper -GC EXAFLEX. It was obtained from a GC Dental Industries Corp in Japan. Rosemary
Our Ohio company is seeking engineering / scientist support for analysis of chemistry and microstructure of solid lubricant and hard coatings used in lubrication of aerospace systems. Work will involve preparing SEM & TEM specimens of thin films and wear scars on steel and ceramic substrates; correlating thin film properties with deposition plasma characteristics; and making recommendations for improving lifetime and performance of such materials in different environments: e.g., vacuum, moist air, high temperature, etc.
It is important that candidates have capabilities in cross-section TEM, analytical TEM, analysis of unique microstructures; and understand TEM of thin films on a fundamental level. It is desirable that the candidate have knowledge of tribological materials and experience with TEM/XTEM of wear tracks.
Contact Ronald Decker - mailto:decker-at-utcdayton.com
{} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} Ronald C. Decker Program Manager Universal Technology Corporation 1270 N FAIRFIELD RD DAYTON OH 45432-2600
Voice (937) 426-8530, Fax (937) 426-7753 (Voice mail is available at my extension, 270)
SEM Fractographs by Svend Engell-Nielson The Technological Institute DK - 2630 Tastrup, Denmark =AE1975
J. Roy Nelson Material Testing Laboratory Pennington, NJ 08534
"Barry Searle (by way of Nestor J. Zaluzec)" wrote:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } } Chaps, Some time ago there was commercially produced a wall cha= rt } (Germany??) that displayed the different types of fracture surfaces (+= a } few errors) as observed using an SEM and which was used as an aid / dis= play } by metallurgists. Does anyone out there have a similar wall chart in } their SEM / Metallurgy lab or perhaps know any of the details of simila= r } types of wall charts. Possibly, it is no longer available?? Thanky= ou } for your help Barry M UNIT UNSW
These materials should be available from any good dental supply agent we have used various polyvinylsiloxane materials: Extrude (polyvinylsiloxane) (Kerr Manufacturing Co, Romulusm MI 48174) and Express (3M Dental Products Division, St Paul, MN 55144- 1000) this latter had a note on the box - U.S. Federal Law restricts this device to sale by or on the order of a dental professional.
Hope this helps
Ian
} Dear Listers, } I'm lookling for a commercial source in the US } for a low viscosity vinyl silicone (dental impression material). } I found a commercial product in a paper -GC EXAFLEX. It } was obtained from a GC Dental Industries Corp in Japan. } Rosemary } } }
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
Hi all, We have just a biopsy arrive in our lab, which is a platelet sample in paraffin wax. Now we were wondering, do we just de-paraffinize as usual in xylene, but spinning it down between each change? Then spinning it down at each re-hydration step til into buffer where when we can embed it into agarose or something like that?
We havent had one of these before, and as usual, would hate to loose what precious little we have been given!
Would appreciate hearing from people who have done this before,
Ta!!
Rich.
----------------------------------------------------------------------- Richard Lander Electron Microscopist South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254 mailto:richard.lander-at-stonebow.otago.ac.nz http://www.otago.ac.nz/anatomy/emunit/ ------------------------------------------------------------------------
I agree with this and would like to add another concern. In library school, my rare books professor would always say, "Bibliographic description forms the basis of textual criticism", meaning that you have to have a way of identifying the item that you are commenting on so that all scholarly discussion of the publication is 'on the same wavelength'. So if you have links to publications on people's own servers and they decide to make a few updates here and there, you have to have a way of identifying the different versions of the documents and this method should be standardized. Having the publications go through a centralized review process and having static versions available on a server or group of mirror servers is preferable, I think.
I don't know what the NIH project is going to be like, but I'm looking forward to it, considering what a success Medline is. Another project to watch for is SPARC http://www.arl.org/sparc
Electronic publishing might provide the opportunity to come up with solutions to the problems of lag time and undue cost. However, electronic publishing in and of itself is not the answer (some ejournals are very expensive and e-publications can suffer massive delays just like print).
Back to the original subject of the post: an online site for product reviews and information sounds like a very good idea.
PS - An entertaining account of the "cold fusion" debacle (as well as "Biosphere II" and others) can be found in the book _Yes, We Have No Neutrons..._
On Thu, 16 Sep 1999, Michael Bode wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Let me throw in a few thoughts... } } While I agree that traditional publication procedures are slow and } cumbersome, they do provide an important service: reviewing. Without } review, the quality of the publications will quickly degrade to a point } that makes publishing useless. Just check out the physics newsgroups on } the net. When I last looked there, there were a number of whackos who } were arguing for the weirdest ideas (Earth core is made of strawberry } jelly kind of thing). I am afraid, that these people would quickly take } over any unreviewed publishing site, and make it completely useless for } scientific publications. } } Also look at the scientific journals: the ones with the strictest } screening are in general respected the most. } } I would think, that in order to provide a value to the science } community, it is a good idea to go digital on the publications WITHOUT } sacrificing the quality of the publications, i.e., without giving up } reviewing. } } I remember a few years back there was a big uproar in the physics } community about some research that was published in the New York Times } before it made it into a scientific journal (I forgot, what exactly it } was. Cold Fusion??). } } Just my thoughts... } } 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: Gordon Vrololjak[SMTP:GVRDOLJA-at-NATURE.BERKELEY.EDU] } } Sent: Wednesday, September 15, 1999 11:50:57 AM } } To: Gordon Couger } } Cc: Sara Miller; Paul Webster; MSA listserver submission } } Subject: Re: Recommnedations } } Auto forwarded by a Rule } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 totally agree with this, } Journal review is taking far too long now. Scientists should really put } more effort in reviewing their co-workers paper submissions. Papers } sitting on someone's desk for six months without review is unacceptable. } I personally try to finish a review once I receive it within two weeks. } Many people asked to review papers often never reply to the publishers, } which I think is a serious offence in the scientific field. } } I know the NIH is considering doing rapid article publication on their } website. After screening of the articles, they are placed onto the web } for anyone to download or read. I think there are other journals going } this route as well. It is the communication method of the future. I } think that publications will go this way. More respected online } journals } will have extensive peer review and screening. Other rapid publication } journals will have less, and still be immensely useful in being able to } do } online searching for specific information. } } I have a lot of experience in programming, web design, and system } administration. I am interested in forming a rapid publication web site } for the scientific community and will readily offer my skills and time. } Gordon Vrdoljak. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ } \/\/\ } Gordon Ante Vrdoljak 1 Cyclotron Road } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja MS90-1116 } GAVrdoljak-at-lbl.gov Ernest Orlando } phone (510) 495-2829 Lawrence } Berkeley } fax (510) 486-7797 National } Laboratory } cell (510) 290-6793 Berkeley CA } 94720 } } On Tue, 14 Sep 1999, Gordon Couger wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } I have the space to keep the data base. I don't have the expertise } } to edit it. It is a pretty good link. It is not as fast as the best } but } } the congestion is low enough it makes up for it most of the time. } } } } I will consider any form of internet publication. I am looking for } } a small circulation juried journal. I am primarily interested in } } increasing the speed of information distribution and reducing } } the cost. } } } } My experience with some of my own publications where it took } } a year to get it published after acceptance and $700 bucks for } } reprints and they kept the copyright left a bad taste in my mouth. } } } } If we are going to increase the rate of progress in technology we } } need to get the time to publication down as low as possible. I think } } it should be possible to get a non controversial jurried article out } } in two months and letters out in hours. } } } } I am interested in publishing only. I am no editor. All copyrights } } stay with the author and no page cost at this point. The cost of } } publication on the internet is very low. My cost } } for keeping a box on the net are 50 a month and hardware } } replacement. $1,000 a year covers the cost including administration } } after the first time set up. So very modest page charges would } } be very attractive to and ISP. My interest is getting it started. } } } } My cost will go up with a lot of net acess. But not much. } } } } Gordon } } Gordon Couger } } 624 Cheyenne } } Stillwater, OK 74075 } } 405 624 2855 GMT - 6:00 } } -----Original Message----- } } } From: Sara Miller {saram-at-duke.edu} } } To: Paul Webster {pwebster-at-mailhouse.hei.org} } } Cc: MSA listserver submission {Microscopy-at-Sparc5.Microscopy.Com} } } Date: Tuesday, September 14, 1999 4:22 PM } } Subject: Re: Recommnedations } } } } } } } } ----------------------------------------------------------------------- } - } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } ----------------------------------------------------------------------- }
Hi All I have recently had my EDS detector repaired and upgraded from a Be window to an ATW window. I am in the process of recalibrating standards for use in ZAF/PB on an AN 10000 system. I have successfully calibrated elements from Na upwards. I have succesfully created profiles for C , N and O but when trying to calibrate the standard to derive FST and RST the system crashes with errors. Is anyone sucessfully using ZAF/PB with a light element detector? Or has anyone any idea why the system might object to light element quant? Many thanks
Chris
Chris Gilpin Experimental Officer Biological Sciences EM Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 0161 275 5170 Fax +44 0161 275 5171 http://www.empgu.man.ac.uk
Dear All, I enclose details of a post-doc vacancy at the University of Barcelona, Spain. The starting date would be about April-May 2000, so we have extended the deadline for applications. Any one interested please reply directly to paqui-at-el.ub.es and/or send applications and a CV by mail before 30 November 1999.
Kind regards
F. Peir=F3
**************************************************************************= ** Laboratory: Electronic Materials and Engineering, Department of Electronics, University of Barcelona.
Duration: 12-18 months, starting April-May 1999.
Raising the subject yet again:
We are seriously considering purchasing a scanner for negatives ranging from 35mm (from optical mic., SEM, Philips 310) and plates about 3x4" (from Philips CM20).
I've looked through the archive, and come up with recommendations for:
Agfa Duoscan (web page kindly provided) Polaroid Sprintscan 45 Nikon LS-4500 for $6500 ** Minolta Dimage Scan Multi for $2500 ** UMAX (generally)
** (from Publishing Perfection)
People over this side of the pond have generally recommended Heidelberg "Saphir" or similar.
Could anybody give me up-to-date recommendations, in particluar addressing the question as to whether it makes sense to have (1) a dedicated scanner for negatives and one for A4 (2) one to do everything.
Solution 1 recommends itself, as the prints, etc, to be scanned might be sometimes be a little grubby from storage.
Pointers to WEB SITES would be much appreciated.
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 | +------------------------------------------------------------------------+
Robin Cross emailed the diagram. We have the unit fixed. Cordial thanks to all who replied.
Amazing place. Amazing people.
Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories (MIL) and Department of Biology University of California at San Diego
address: 1500 Bonner Hall, University of California at San Diego 9500 Gilman Drive, La Jolla, CA 92093-0368 telephone - lab: 8585342484 telephone - office: 8588223373 pager: 8586161420 fax: 8588223715 email: mmm-at-biomail.ucsd.edu www site: http://mil.ucsd.edu
Your AS 325 Microtome was produced by Anglia Scientific who were taken over by Shandon who in turn became part of Life Sciences International. The last address I have for them is as follows:
Life Sciences International (Europe) Ltd 93-96 Chadwick Road Astmoor Runcorn Cheshire WA7 1PR Tel 01928 566611 Fax 01928 565845
Many microtomes have drifted under the bridge since these changes but if you have any problems please contact me and I may be able to trace something,
Regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, UK
Postdoctoral Position to Develop Advanced Microscopy Capabilities in a Microscopy User Center
The Materials Analysis User Center (MAUC) at the Oak Ridge National Laboratory in Oak Ridge, Tennessee, is seeking a postdoctral candidate to perform research to facilitate the use of electron microscopes remotely, and to improve TEM resolution using electron beam monochromators and hardware aberration correctors.
There are two primary areas that the person selected for this position will be expected to contribute. First, this center and the Oak Ridge National Laboratory have strongly supported the concept of a virtual electron microscopy laboratory over the last years and will continue to move in that direction. Second, in support Department of Energy research programs aimed at reducing both gaseous and particulate emissions from gasoline and diesel engines, the ultimate in TEM atomic resolution (at intermediate voltages) for nano-particles and other materials is a critical need for this center.
The successful candidate should have a PhD in physics (although other disciplines will be considered, depending upon overall qualifications). The candidate must have a strong background in all of the following: (1) electron optics (preferably in the design and use of electron monochromators and aberration correctors), (2) programming skills including C and C++ (Java experience is desired), and (3) hands-on experience with TEM and STEM instrumentation. The researcher will work with many DOE contractors, industrial and university researchers, including some as a part of DOE-ORNL user programs; thus he or she should enjoy collaborating with others. This position is for one year, extendable to two.
ORNL, a multipurpose research laboratory managed by Lockheed Martin Energy Research Corporation for the U.S. Department of Energy, is an equal opportunity employer committed to building and maintaining a diverse work force.
Please send curriculum vitae and bibliography to:
Ted Nolan Manager, Materials Analysis User Center High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 Oak Ridge, TN 37831-6064
Also very narrow groups could get wider representation. Cross feild porject have been the best project I have worked on.
Gordon
} } I agree with this and would like to add another concern. } In library school, my rare books professor would always say, } "Bibliographic description forms the basis of textual criticism", } meaning that you have to have a way of identifying the item that you are } commenting on so that all scholarly discussion of the publication is } 'on the same wavelength'. So if you have links to publications on people's } own } servers and they decide to make a few updates here and there, you have to } have a way of identifying the different versions of the documents and this } method should be standardized. Having the publications go through a } centralized review process and having static versions available on a } server or group of mirror servers is preferable, I think. } } I don't know what the NIH project is going to be like, but I'm looking } forward to it, considering what a success Medline is. Another project to } watch for is SPARC } http://www.arl.org/sparc } } Electronic publishing might provide the opportunity to come up } with solutions to the problems of lag time and undue cost. However, } electronic publishing in and of itself is not the answer (some ejournals } are very expensive and e-publications can suffer massive delays just like } print). } } Back to the original subject of the post: } an online site for product reviews and information sounds like a very good } idea. } } PS - An entertaining account of the "cold fusion" debacle } (as well as "Biosphere II" and others) can be found in the book } _Yes, We Have No Neutrons..._ } } On Thu, 16 Sep 1999, Michael Bode wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Let me throw in a few thoughts... } } } } While I agree that traditional publication procedures are slow and } } cumbersome, they do provide an important service: reviewing. Without } } review, the quality of the publications will quickly degrade to a point } } that makes publishing useless. Just check out the physics newsgroups on } } the net. When I last looked there, there were a number of whackos who } } were arguing for the weirdest ideas (Earth core is made of strawberry } } jelly kind of thing). I am afraid, that these people would quickly take } } over any unreviewed publishing site, and make it completely useless for } } scientific publications. } } } } Also look at the scientific journals: the ones with the strictest } } screening are in general respected the most. } } } } I would think, that in order to provide a value to the science } } community, it is a good idea to go digital on the publications WITHOUT } } sacrificing the quality of the publications, i.e., without giving up } } reviewing. } } } } I remember a few years back there was a big uproar in the physics } } community about some research that was published in the New York Times } } before it made it into a scientific journal (I forgot, what exactly it } } was. Cold Fusion??). } } } } Just my thoughts... } } } } 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: Gordon Vrololjak[SMTP:GVRDOLJA-at-NATURE.BERKELEY.EDU] } } } Sent: Wednesday, September 15, 1999 11:50:57 AM } } } To: Gordon Couger } } } Cc: Sara Miller; Paul Webster; MSA listserver submission } } } Subject: Re: Recommnedations } } } Auto forwarded by a Rule } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 totally agree with this, } } Journal review is taking far too long now. Scientists should really put } } more effort in reviewing their co-workers paper submissions. Papers } } sitting on someone's desk for six months without review is unacceptable. } } I personally try to finish a review once I receive it within two weeks. } } Many people asked to review papers often never reply to the publishers, } } which I think is a serious offence in the scientific field. } } } } I know the NIH is considering doing rapid article publication on their } } website. After screening of the articles, they are placed onto the web } } for anyone to download or read. I think there are other journals going } } this route as well. It is the communication method of the future. I } } think that publications will go this way. More respected online } } journals } } will have extensive peer review and screening. Other rapid publication } } journals will have less, and still be immensely useful in being able to } } do } } online searching for specific information. } } } } I have a lot of experience in programming, web design, and system } } administration. I am interested in forming a rapid publication web site } } for the scientific community and will readily offer my skills and time. } } Gordon Vrdoljak. } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ } } \/\/\ } } Gordon Ante Vrdoljak 1 Cyclotron Road } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja MS90-1116 } } GAVrdoljak-at-lbl.gov Ernest Orlando } } phone (510) 495-2829 Lawrence } } Berkeley } } fax (510) 486-7797 National } } Laboratory } } cell (510) 290-6793 Berkeley CA } } 94720 } } } } On Tue, 14 Sep 1999, Gordon Couger wrote: } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -----------------------------------------------------------------------. } } } } } } } } } I have the space to keep the data base. I don't have the expertise } } } to edit it. It is a pretty good link. It is not as fast as the best } } but } } } the congestion is low enough it makes up for it most of the time. } } } } } } I will consider any form of internet publication. I am looking for } } } a small circulation juried journal. I am primarily interested in } } } increasing the speed of information distribution and reducing } } } the cost. } } } } } } My experience with some of my own publications where it took } } } a year to get it published after acceptance and $700 bucks for } } } reprints and they kept the copyright left a bad taste in my mouth. } } } } } } If we are going to increase the rate of progress in technology we } } } need to get the time to publication down as low as possible. I think } } } it should be possible to get a non controversial jurried article out } } } in two months and letters out in hours. } } } } } } I am interested in publishing only. I am no editor. All copyrights } } } stay with the author and no page cost at this point. The cost of } } } publication on the internet is very low. My cost } } } for keeping a box on the net are 50 a month and hardware } } } replacement. $1,000 a year covers the cost including administration } } } after the first time set up. So very modest page charges would } } } be very attractive to and ISP. My interest is getting it started. } } } } } } My cost will go up with a lot of net acess. But not much. } } } } } } Gordon } } } Gordon Couger } } } 624 Cheyenne } } } Stillwater, OK 74075 } } } 405 624 2855 GMT - 6:00 } } } -----Original Message----- } } } } From: Sara Miller {saram-at-duke.edu} } } } To: Paul Webster {pwebster-at-mailhouse.hei.org} } } } Cc: MSA listserver submission {Microscopy-at-Sparc5.Microscopy.Com} } } } Date: Tuesday, September 14, 1999 4:22 PM } } } Subject: Re: Recommnedations } } } } } } } } } } } } ----------------------------------------------------------------------- } } - } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } ----------------------------------------------------------------------- } } } } } }
Robert, I use a Linotype-Hell Saphir Ultra (purchased before Linotype-Hell was bout out by Heidelberg Press). Very nice machine, but: 1) There is no provision (at least in the version of software that I have) for setting the darkest and brightest parts of the image in preview. You just have to go with whatever automatic correction the software uses. 2) I have had some reliability problems, the power supply died about three months after I bought it and it still has an intermittent fault in transparency mode where the light source above and CCD array below do not align correctly - giving poor quality images.
I used an Agfa some years ago which had (1) and it was much easier to use for scanning TEM negatives. I don't know whether my problems (2) are just for my machine or a reflection on the machines generally. I believe that the CCD and mechanical parts are from UMAX (also the case for Agfa)? I also believe that the maximum OD and optical (not interpolated) resolution has increased somewhat since I got my scanner about 18 months ago.
Despite the niggles, it is a very good and generally reliable machine which must have scanned about 2000 pictures in the last 18 months. It is used for everything from TEM negatives and X-ray radiographs to taking pictures from old reports (and of course holiday/wedding/old family photos out of hours!)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Raising the subject yet again: } } We are seriously considering purchasing a scanner for negatives ranging } from 35 mm (from optical mic., SEM, Philips 310) and plates about 3x4" } (from Philips CM20). } } I've looked through the archive, and come up with recommendations for: } } Agfa Duoscan (web page kindly provided) } Polaroid Sprintscan 45 } Nikon LS-4500 for $6500 ** } Minolta Dimage Scan Multi for $2500 ** } UMAX (generally) } } ** (from Publishing Perfection) } } People over this side of the pond have generally recommended Heidelberg } "Saphir" or similar. } } Could anybody give me up-to-date recommendations, in particular addressing } the question as to whether it makes sense to have } (1) a dedicated scanner for negatives and one for A4 } (2) one to do everything. } } Solution 1 recommends itself, as the prints, etc., to be scanned might be } sometimes be a little grubby from storage. } } Pointers to WEB SITES would be much appreciated. } } 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 | } +------------------------------------------------------------------------+ } }
I have an EDS system with a 30 mm premium detector which had original spec's of 136 eV. The crystal pack needed repair and was replaced. The resolution now is 142 eV. I need to decide a course of action. Here are my options.
1. Keep the repaired 30 mm detector at 142 eV and be back up and running. 2. Here the vendor replace the crystal pack until it meets the 136 eV spec, which will take time and I might be down for a while. 3. Go to a 10 mm detector that I am told they can insure a resolution of 136 or better.
Question for the group, if I can live with the down time should I hold out for the 30 mm detector at 136eV?
Michael Ingram Rodel, Inc. 451 Bellevue RD Newark, DE 19713 Phone: 302.366.0500, ext. 2545 Fax: 302.455.1124
Does anyone in the Rochester, NY area have a JEOL 5800 SEM (or similar conventional SEM)? I will be in that region and I must test some equipment, so I need to gain access to a SEM similar to mine for a short period.
Please contact me off-line if this is possible, and for further info. Marisa Ahmad
I am rather surprised this a.m. to arrive at the lab to find that the = blocks I've embedded in LR White are not polymerized. This is not a = technique we routinely use. I followed the schedule for EM ICC, tissues = are .25-.5mm, no OSO4, ETOH dehydration to 95%, several infiltration = changes over hours and overnight, oven calibrated to 50 degrees, gelatin = capsules totally filled and capped, in the oven since Friday. Help -- What's going wrong?? What to do now ?? Linda Fox lfox1-at-wpo.it.luc.edu
I have a Gatan 600 Duo Mill Ion Miller of the 1985 vintage.
The Thermocouple gauge controller is malfunctioning and will not allow the difusion pump to turn on. It is the gauge on the right side of the miller. The gauge model number is CVH-3.
If anyone has the circiuit diagram of the interior of the controller I would appreciate a copy.
It can be faxed to (905) 521-2773. When faxing please use fine resolution since detail on the diagram is rather small.
Thanks in advance
Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
Wow, 136 eV with a 30 mm detector is exceptional, and I can see why you are a little bummed out that the repair came back 6 eV worse. But is that 6 eV important to you? I would guess that in practice you won't see much difference. Personally I would rather have the big detector.
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
Mike Ingram wrote:
:I need opinions. : :I have an EDS system with a 30 mm premium detector which had original spec's :of 136 eV. The crystal pack needed repair and was replaced. The resolution :now is 142 eV. I need to decide a course of action. Here are my options. : :1. Keep the repaired 30 mm detector at 142 eV and be back up and :running. :2. Here the vendor replace the crystal pack until it meets the 136 eV :spec, which will take time and I might be down for a while. :3. Go to a 10 mm detector that I am told they can insure a resolution :of 136 or better. : :Question for the group, if I can live with the down time should I hold out :for the 30 mm detector at 136eV? : :Michael Ingram :Rodel, Inc. :451 Bellevue RD :Newark, DE 19713 :Phone: 302.366.0500, ext. 2545 :Fax: 302.455.1124
Hello every one Here is a quandary for you If you HAD to keep tissue (in this case 50 micron vibratome sections) at either the primary fixation step (2% PF and 2% Glut), or in a buffer at 4 degrees C after the primary fix. Which would be the most stable over several weeks time? Lets assume they will be looking for general ultrastructure. I know there are lots of variables but I'm looking for an "in general" answer.
I get request from people to do EM but I can not get their sample processed into resin at the time, so here I have a sample to store or in other cases they want to store it until they know if they will need the EM.
In the above experiment, another "EM" investigator told them to store the sample in a phosphate buffer at 4 degree. Due to the number of samples I won't be able to process them all at once so I'm not sure whether to leave them the way they are or transfer them back into fix. I personally think it's best to leave the sample in the primary fix at 4 degree. In the Path EM lab we used to keep samples this way, at room temp, and the tissue showed no discernible loss of ultrastructure even when stored a month or two. Thanks for your advice.
At 09:51 AM 9/20/99 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
****************************** If you post fix in osmium, you might carry it to the buffer rinse after the osmium step and store it in buffer at 4C. The tissue isn't fixed completely until it has been osmicated. This way both proteins and lipids will be well preserved.
Joiner Cartwright, Jr., Ph.D. Assistant Professor of Pathology Baylor College of Medicine Houston, Texas U.S.A.
I had the similar problem once before. I checked the LR white resin (which is newly purchased) withOUT sample and polymerized overnight at 50 or 70 C. It did NOT polymerize either. I called the supplier (EMS) and they sent me a new one. It worked fine?!
Linda Fox wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am rather surprised this a.m. to arrive at the lab to find that the blocks I've embedded in LR White are not polymerized. This is not a technique we routinely use. I followed the schedule for EM ICC, tissues are .25-.5mm, no OSO4, ETOH dehydration to 95%, several infiltration changes over hours and overnight, oven calibrated to 50 degrees, gelatin capsules totally filled and capped, in the oven since Friday. } Help -- What's going wrong?? What to do now ?? } Linda Fox } lfox1-at-wpo.it.luc.edu
Zhaojie Zhang Program in Molecular and Cell Biology Oklahoma Medical Research Foundation 825 NE 13th Street Oklahoma City, OK 73104
Dear Michael and Mark, 136 eV is not so exceptional if you are willing to put up with the long process times and high dead time %. If you are getting 142 eV with the longest process time, it might be worth it to wait for a better detector. I don't think the resolution is as important as the loss in counts and time you will incurr. We don't set our detector at the highest resolution so that we can get more counts per real time. It makes a difference for X-ray mapping where every nanosecond counts. You may also twist their arm into giving you a loaner. Ciao for now, Ken
} Dear Michael, } } Wow, 136 eV with a 30 mm detector is exceptional, and I can see } why you are a little bummed out that the repair came back 6 eV } worse. But is that 6 eV important to you? I would guess that } in practice you won't see much difference. Personally I would } rather have the big detector. } } } 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 } } Mike Ingram wrote: } } :I need opinions. } : } :I have an EDS system with a 30 mm premium detector which had original spec's } :of 136 eV. The crystal pack needed repair and was replaced. The resolution } :now is 142 eV. I need to decide a course of action. Here are my options. } : } :1. Keep the repaired 30 mm detector at 142 eV and be back up and } :running. } :2. Here the vendor replace the crystal pack until it meets the 136 eV } :spec, which will take time and I might be down for a while. } :3. Go to a 10 mm detector that I am told they can insure a resolution } :of 136 or better. } : } :Question for the group, if I can live with the down time should I hold out } :for the 30 mm detector at 136eV? } : } :Michael Ingram } :Rodel, Inc. } :451 Bellevue RD } :Newark, DE 19713 } :Phone: 302.366.0500, ext. 2545 } :Fax: 302.455.1124
I have an old JEOL JAMP-30 Auger Microprobe, and the monitor for the PDP 11/70 microcomputer (the computer that runs the auger software) has just died. The monitor is a Nissho Electronics Pictus 80I. Does anyone have a replacement for sale (or donation) or any idea where I might find one? Any assistance is greatly appreciated.
Brian M. Robin Research Assistant Analytical Instrumentation Facility North Carolina State University
The Alberta Research Council -Vegreville site has a Hitachi H600 TEM (with STEM attachment and Kevex 700 microanalysis system) available for the cost of moving it from our site to yours. The TEM requires a new high tension cable to be functional.
If anyone is interested in the entire unit (or maybe just parts of it) please contact either Rosemary Harris (rosemary-at-arc.ab.ca or telephone 780-632-8451) or Arlene Oatway (arlene-at-arc.ab.ca or telephone 780-632-8454).
Vegreville is located about 60 miles east of Edmonton in Alberta, Canada.
How you store it depends on your ultimate goal. If you're not going to do immunolabeling, store it in glut/paraform. These don't fix some structures well and are somewhat reversible; thus, storing in buffer for long times afterward could leach out contents. It would be even better if you could process tissue through osmium and then store in buffer.
If you're going to do IEM, don't store tissue long periods in fix, unless you know that your antigen survives this treatment. Some do; some don't.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We have a Kevex 8000 EDS spectrometer we do not need. The detector is a three year old thin window Quantum detector. The analyzer is probably ten years old. The Kevex detector has been horizontally mounted on an Amray 1000A scanning electron microscope, with which we are parting.
In addition, we have an Edax 9100 EDS system we do not need.
We are open to offers on both the Kevex 8000 system and the Edax 9100 system. If someone wanted both the Kevex 8000 system and the Amray 1000A SEM, that could be arranged.
I have nine un-opened 1.5 GB rewritable optical disks (double-sided) that I will never use:
Panasonic # LM-R1500A
Originally purchased for use in an Optical Access International optical disk drive.
If anyone is certain they can use these disks, please contact me.
Peter Guthrie Department of Neurobiology & Anatomy University of Utah School of Medicine 50 N Medical Drive Salt Lake City, UT 84132 (801) 581-8336 (801) 581-4233 fax Peter.Guthrie-at-hsc.utah.edu
Hello, My company is interested in obtaining an EDAX system for our JEOL T220A SEM. As this system is used on a limited basis, purchasing an entirely new system is not cost effective. Can anyone suggest resources for finding used equipment, or know of any directly? Thank you in advance.
Dysktra (spelling? this is off the top of my head) in his EM text showed micrographs of monkey kidney that had been in Trump's fix for 4 years and didn't look all that bad. (Trump's is basically the same as what you're using, slightly different %s of glut & formaldehyde.) I'd keep the specimens in fix in the frig.
Phil
} Hello every one } Here is a quandary for you } If you HAD to keep tissue (in this case 50 micron vibratome sections) at } either the primary fixation step (2% PF and 2% Glut), or in a buffer at 4 } degrees C after the primary fix. Which would be the most stable over } several weeks time? Lets assume they will be looking for general } ultrastructure. } I know there are lots of variables but I'm looking for an "in general" } answer. } } I get request from people to do EM but I can not get their sample processed } into resin at the time, so here I have a sample to store or in other cases } they want to store it until they know if they will need the EM. } } In the above experiment, another "EM" investigator told them to store the } sample in a phosphate buffer at 4 degree. Due to the number of samples I } won't be able to process them all at once so I'm not sure whether to leave } them the way they are or transfer them back into fix. I personally think } it's best to leave the sample in the primary fix at 4 degree. In the Path } EM lab we used to keep samples this way, at room temp, and the tissue } showed no discernible loss of ultrastructure even when stored a month or } two. Thanks for your advice. } } Rick Vaughn } RLVAUGHN-at-UNMC.EDU
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Dear All, Greetings of the day. Last year I had registered to attempt the MSA certification exam. Logistical problem of taking the exam from outside the US prevented me, hence, I could not take it in the previous cycles. Could any of you provide me with the new dates for the upcomming two cycles of this exam. Then again, I shall follow it up with the chair of the exam and the businessoffice of the MSA. Thanks
Mohammed Yousuf Dept. of Zoology King Saud University POB 2455 Riyadh 11451 Saudi Arabia
Hi everyone, If you know anybody that could be interested by the following job offer, please forward this email. Thanks Cecile
ps : don't sent any enquiries to me , send them to Professor J.S. Abell or Dr. T.W. Button School of Metallurgy and Materials University of Birmingham Birmingham B15 2TT
School of Metallurgy and Materials in Collaboration with Oxford Instruments Plc Research Fellowship in the Development of Superconducting Coated Tapes Applications are invited for a post-doctoral research position to work on the fabrication and characterisation of bi-axially textured YBCO films on metallic substrates. The post is supported by funding from one of the leading international industrial companies involved in the development of practical superconducting wires and tapes. The successful candidate will join an established interdisciplinary research group at the University, but will benefit from close interaction with the company both in the UK and US. Some experience of thick or thin film technology of HTC materials would be an advantage, with expertise in pulsed laser deposition being particularly attractive. The appointment will be for two years initially. Interested candidates should have a PhD in materials science or related subject.
The vacancy will be advertised formally in a short time, but informal enquiries should be sent to
Professor J.S. Abell or Dr. T.W. Button School of Metallurgy and Materials University of Birmingham Birmingham B15 2TT
J.S. Abell-at-bham.ac.uk ( 0121 414 5168 ) T.W. Button-at-bham.ac.uk ( 0121 414 5237 ) ****************************** Dr. Cecile Prouteau School of Metallurgy & Materials The University of Birmingham Edgbaston, Birmingham B15 2TT UK E-mail : c.prouteau-at-bham.ac.uk tel : (UK=44)- (0)121 414 5170 fax : (UK=44)- (0)121 414 5232
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I saw the post on LR white and noticed the no OsO4 part of the protocol. I'm having problems with some LR white just now and I'm wondering if its because the tissue (brown fat in this case) was Osmicated. The block seems hard enough except where the tissue is and around there its like rubbery butter. Interestingly enough the non osmicated blocks seem fine if maybe a little too hard. Any ideas out there?
Rod Nicholls Head Technician EM Unit University of Ottawa
Rick, its not a quandary for me. Microscopists unwilling to put in any additional effort to assure good EM results - if that course was to be subsequently chosen, pay later for their omissions. The rules are simple: For best results, tissues should be post-fixed in osmium as soon as possible. Fully fixed tissues may be stored in the cold in either buffer or a low, say 50% alcohol.
Sabatini advocated that GA fixed tissue could be held for six months in buffer and then postfixed, but only some tissues will give good results after that delayed fixation. Similarly, tissues stored in GA (forget storing in osmium!), will continue crosslinking and at high powers show a more granular texture. Sjostrand thought that he could avoid this false granularity completely, by fixing single cells for under a minute in GA. He had hoped to retain cellular details to a resolution well below 1nm. This is ancient stuff now and you may not obtain true details at such resolution from tissue sections, but storing tissues in fixative is not a good idea. 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 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, September 21, 1999 12:52 AM, "rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com [SMTP:"rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com] wrote: } } Hello every one } Here is a quandary for you } If you HAD to keep tissue (in this case 50 micron vibratome sections) at } either the primary fixation step (2% PF and 2% Glut), or in a buffer at 4 } degrees C after the primary fix. Which would be the most stable over } several weeks time? Lets assume they will be looking for general } ultrastructure. } I know there are lots of variables but I'm looking for an "in general" } answer. } } I get request from people to do EM but I can not get their sample processed } into resin at the time, so here I have a sample to store or in other cases } they want to store it until they know if they will need the EM. } } In the above experiment, another "EM" investigator told them to store the } sample in a phosphate buffer at 4 degree. Due to the number of samples I } won't be able to process them all at once so I'm not sure whether to leave } them the way they are or transfer them back into fix. I personally think } it's best to leave the sample in the primary fix at 4 degree. In the Path } EM lab we used to keep samples this way, at room temp, and the tissue } showed no discernible loss of ultrastructure even when stored a month or } two. Thanks for your advice. } } Rick Vaughn } RLVAUGHN-at-UNMC.EDU }
I am not sure if this is the right place, but maybe somebody can help.... A colleague of mine send this to me since I operate our EM's, but before I run a serious search:
"We do laser marking of anodised aluminum, and have a problem. Some mark extremely well and other samples mark extremely poorly. I need to find out the reason for this, and have a number of things I would like to test :
1) the difference between different aluminum alloys 2) the difference between anodised thicknesses 3) the difference between anodised colors 4) the effect of surface finish.
Can you help with any of these ...or suggest some technique do to measure the anodised thickness etc etc. "
Any help would be much appreciated.
Many thanks Sara
Sara Prins CSIR-National Metrology Laboratory PO Box 395 Pretoria South Africa Tel +27 12 841 3974 Cell +27 83 2972643 Fax +27 12 841 2131 e-mail sprins-at-csir.co.za http://nml.csir.co.za
:I need opinions. : :I have an EDS system with a 30 mm premium detector which had original spec's :of 136 eV. The crystal pack needed repair and was replaced. The resolution :now is 142 eV. I need to decide a course of action.} }
You probably paid more to have a 136 detector, 142's are pretty much off the reject pile IMHO.
We have also been having trouble with LR White, specifically with embedding plant cells and tissue. We have tried dehydrating though 70% ethanol, then through 100%. We have increased infiltration times, played with embedding times and temperatures, and tried different bottles of resin. We get sections that fall apart in the boats and resembled tattered napkins on the grids. The tissues and cells have been processed for immunolabeling and the labeling is working perfectly on the surviving portions of the sections, but the sections themselves look awful.
Our other resins are all working fine. So we join the ranks of the puzzled ones on this question. Any help would be appreciated.
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine Bldg. University of Missouri Columbia, MO 65211 (573)882-8304 http://www.biotech.missouri.edu/emc/
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I am rather surprised this a.m. to arrive at the lab to find that the } } blocks I've embedded in LR White are not polymerized. This is not a } } technique we routinely use. I followed the schedule for EM ICC, tissues } } are .25-.5mm, no OSO4, ETOH dehydration to 95%, several infiltration } } changes over hours and overnight, oven calibrated to 50 degrees, gelatin } } capsules totally filled and capped, in the oven since Friday. } } Help -- What's going wrong?? What to do now ?? } } Linda Fox } } lfox1-at-wpo.it.luc.edu } } I saw the post on LR white and noticed the no OsO4 part of the protocol. } I'm having problems with some LR white just now and I'm wondering if its } because the tissue (brown fat in this case) was Osmicated. The block seems } hard enough except where the tissue is and around there its like rubbery } butter. Interestingly enough the non osmicated blocks seem fine if maybe a } little too hard. Any ideas out there? } } Rod Nicholls } Head Technician } EM Unit } University of Ottawa } } } } Hi,
Osmicated fat or even poorly osmicated fat interferes with polymerization of epoxies. This became absolutely clear to me when I tried to embed some M and Ms for paperweights. The shell cracked, the cocoa butter ran out and the area around it stayed gooey. (I had to design a special formulation which would embed M and Ms) It is possible that the same applies to the acrylics. I have done a lot of studying of Acrylics and have never seen anywhere that fat does interfere. Try dehydration to 100% and see if there is any improvement. Are you infiltrating with vigor? A number of changes, etc. Another possibility is, of course, that the fat takes up large quantities of osmium which forms a barrier to infiltration. This we see regularly in nerve bundles both in epoxies and acrylics.
If the problem does not solve itself, let me know, and I can give you some places where you might get help.
Hildy Crowley Sr. Electron Microscopist University of Denver {hcrowley-at-du.edu}
What kind of elements are typically present in your samples? If you don't routinely analyze for Cd, In, Sn and Sb, or for Pb and Bi, I think you could get along quite well with your resolution at 142 eV. The primary need for the improved resolution would be for resolving peak overlaps, although it may also provide a slight improvement in detectable limit.
Hopefully you can get some adjustment to the repair price for not restoring original performance.
Warren
At 09:29 AM 9/20/1999 -0400, "Ingram, Mike" wrote: } } I need opinions. } } I have an EDS system with a 30 mm premium detector which had original spec's } of 136 eV. The crystal pack needed repair and was replaced. The resolution } now is 142 eV. I need to decide a course of action. Here are my options. } } 1. Keep the repaired 30 mm detector at 142 eV and be back up and } running. } 2. Here the vendor replace the crystal pack until it meets the 136 eV } spec, which will take time and I might be down for a while. } 3. Go to a 10 mm detector that I am told they can insure a resolution } of 136 or better. } } Question for the group, if I can live with the down time should I hold out } for the 30 mm detector at 136eV? } } Michael Ingram } Rodel, Inc. } 451 Bellevue RD } Newark, DE 19713 } Phone: 302.366.0500, ext. 2545 } Fax: 302.455.1124
Thanks again!! The problem may be that there was no catalyst added to the resin...(one should always ask the obvious questions when novices are about ... Thank you) I re-read all the proceedures and nowhere does it state to add the benz. perox. to the resin. I assumed that the b.p. was needed for the cold cure method, where excess heat might be generated, so I did not add any. I reinfiltrated the tissues with another bolttle of resin, from EMS, catalyst added by manufacturer, and returned them to the oven. We will see!! Thanks for all the tips. Many called for higher temps. for polymerization. I am wondering if this will hinder immuno work? I will be persuing this technique for one scientist over the next few months and would appreciate all the tips, tricks and protocols you can spare. Thanks, Linda Fox lfox1-at-wpo.it.luc.edu Fax: 1-708-216-3676
I'm wondering if vapor osmication will work on dry sections. I have sections stained for immuno gold and the protocol calls for grids to be floating on pbs during the vapor fix. I'm wondering if I need to have them wet and then have to wash them or can they be done dry and thus save the sections from the rigors of washing and pbs floating. Thanks in advance.
Rod Nicholls Head Technician EM Unit University of Ottawa
Does any information or publication exist that describes or defines the "typical" variability or range that may be observed between analysts for particle counting techniques using LM? Thanks.
Robb Calabro Pharmaceutical Scientist AstraZeneca Pharmaceutical Westborough, MA USA
Did you mix in the benzoyl peroxide catalyst? really thoroughly? it takes a while to mix in properly. I needed LR White in a hurry once and ordered it from Sigma and found out to my dismay that Sigma supplies LR White without the b.p.
To Rod Nicholls: The literature I have says osmium is not compatible (causes focal points of heat).
Rod Nicholls wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I am rather surprised this a.m. to arrive at the lab to find that the } } blocks I've embedded in LR White are not polymerized. This is not a } } technique we routinely use. I followed the schedule for EM ICC, tissues } } are .25-.5mm, no OSO4, ETOH dehydration to 95%, several infiltration } } changes over hours and overnight, oven calibrated to 50 degrees, gelatin } } capsules totally filled and capped, in the oven since Friday. } } Help -- What's going wrong?? What to do now ?? } } Linda Fox } } lfox1-at-wpo.it.luc.edu } } I saw the post on LR white and noticed the no OsO4 part of the protocol. } I'm having problems with some LR white just now and I'm wondering if its } because the tissue (brown fat in this case) was Osmicated. The block seems } hard enough except where the tissue is and around there its like rubbery } butter. Interestingly enough the non osmicated blocks seem fine if maybe a } little too hard. Any ideas out there? } } Rod Nicholls } Head Technician } EM Unit } University of Ottawa
-- Sonia Cawsey McGowan Copley Library, University of San Diego email: scawsey-at-acusd.edu home page: http://www.acusd.edu/~scawsey
I have noticed similar problems with various biolog. tissues and especially with many carbon materials like charcoal, coke, activated carbons, and others. I have always attributed this problem to the oxygen interference issue in the polymerisation of many of the acrylics. It has been my thought that the "carbon" materials and some tissues have enough absorbed O2, or similar chemistry is going on, such that it inhibits LR White polymerisation at the localized interface of the tissue. This is when I give up and use epoxy based embedding (usually Spurrs low viscosity or similar). Brad Huggins BPAmoco EM Lab Naperville, IL
} ---------- } From: Rod Nicholls[SMTP:nicholls-at-uottawa.ca] } Sent: Tuesday, September 21, 1999 10:02 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: LR White polymerization } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I am rather surprised this a.m. to arrive at the lab to find that the } } blocks I've embedded in LR White are not polymerized. This is not a } } technique we routinely use. I followed the schedule for EM ICC, tissues } } are .25-.5mm, no OSO4, ETOH dehydration to 95%, several infiltration } } changes over hours and overnight, oven calibrated to 50 degrees, gelatin } } capsules totally filled and capped, in the oven since Friday. } } Help -- What's going wrong?? What to do now ?? } } Linda Fox } } lfox1-at-wpo.it.luc.edu } } I saw the post on LR white and noticed the no OsO4 part of the protocol. } I'm having problems with some LR white just now and I'm wondering if its } because the tissue (brown fat in this case) was Osmicated. The block seems } hard enough except where the tissue is and around there its like rubbery } butter. Interestingly enough the non osmicated blocks seem fine if maybe a } little too hard. Any ideas out there? } } Rod Nicholls } Head Technician } EM Unit } University of Ottawa } } }
I am learning to using DTSA to analyse EDX spectra collected with a Link Premium 136eV Si(Li) detector with a super atmospheric thin window. The detector is attached to a JEOL 2010F TEM and has a 30mm2 active area.
I would like to generate simulated spectra within DTSA and compare them with actual spectra. The idea is to determine values for the detector's physical parameters that allow's reasonably good agreement between generated and real spectra.
The problem is I have no idea of the approximate layer thicknesses and compositions appropriate for my detector. I would appreciate learning from DTSA users with similar detectors what parameters they are using. I could then treat these values as a starting point for fine tuning my own values.
I look forward to your replies. 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.
Good question. We used to use the 30 mm wafer detector in the SEM because we thought we needed it for extra surface area in x-ray mapping. We now use 10 mm in one SEM and it works fine for our applications. We still have a 30 mm in the system on our 420T (TEM), to maximize counts. If it is TEM or FESEM, there might be some significant benefit to the larger surface. I also will be interested in the opinion of the experts on this one. Brad Huggins BPAmoco
} ---------- } From: Giles, Bill[SMTP:William.Giles-at-TIMET.com] } Sent: Tuesday, September 21, 1999 10:24 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: RE: EDS Question } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } { {Mike Ingram wrote: } } :I need opinions. } : } :I have an EDS system with a 30 mm premium detector which had original } spec's } :of 136 eV. The crystal pack needed repair and was replaced. The } resolution } :now is 142 eV. I need to decide a course of action.} } } } You probably paid more to have a 136 detector, 142's are pretty much off } the } reject pile IMHO. } } } Bill } TIMET }
Only thing I can tell here is that oxigen is toxic to the resin. If you leave it open to air, it will not polymerize. I always overfill gelatin capsule to exclude as much air as possible. It is messy, but I have not found a clean way yet.
Increase infiltration time would not help much as the resin penetrate rather quicker into tissue. Even though the data sheet claims that you can stop at 70% alcohol step. I found you need always dehydrate more than that. Here is what I do after osmication with nervous tissue of 1-3mm size:
dehydrate 50, 70, 95, 100X2, 10 minutes each step
1:1 LR white with 100% Alcohol for 30 minutes on a rotator
Leave in pure LS white for 3hrs to overnight (for convenience only)
Put tissue in fresh LR white in a gelatin capsule. Fill as much as you can and cover it up. Make sure you close it tightly. It is always messy step.
Polymerize at 50oC over for at least 24 hrs. I have not done it for a while now. But I believe if you stop it by taking it out of oven. Then it will not polymerize any more. Put it back to oven will not help.
If no osmication is required, try UV polymerization.
LR white is alway britle and difficult to deal with. You need section a bit thicker than normally do with other resin (e.g. Epon ). I never go below 75nm. Add a bit of grease (YES!) to the boat will help to keep the section together. Use tiny wood picker, touch your "oily face". Dip it into boat. This is enough to keep the sections together.
I am not sure if this helps. But that is how I did it.
Xuejun
On Tue, 21 Sep 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, } } We have also been having trouble with LR White, specifically with embedding } plant cells and tissue. We have tried dehydrating though 70% ethanol, then } through 100%. We have increased infiltration times, played with embedding } times and temperatures, and tried different bottles of resin. We get } sections that fall apart in the boats and resembled tattered napkins on the } grids. The tissues and cells have been processed for immunolabeling and the } labeling is working perfectly on the surviving portions of the sections, but } the sections themselves look awful. } } Our other resins are all working fine. So we join the ranks of the puzzled } ones on this question. Any help would be appreciated. } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine Bldg. } University of Missouri } Columbia, MO 65211 } (573)882-8304 } http://www.biotech.missouri.edu/emc/ } } }
*********************************************************************** Xuejun Sun, Ph.D. Dept. of Experimental Oncology Cross Cancer Institute 11560 University Ave. Edmonton Alberta T6G 1Z2, Canada
Yes Rod, The instructions advise not to use osmium and our extensive online application notes also make that point. Anyway, what is the point of using osmium with LR White. That resin is well suited to immuno and cytochem processing, but these reactions are "killed" by osmium. 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 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
} } } } I am rather surprised this a.m. to arrive at the lab to find that the } } blocks I've embedded in LR White are not polymerized. This is not a } } technique we routinely use. I followed the schedule for EM ICC, tissues } } are .25-.5mm, no OSO4, ETOH dehydration to 95%, several infiltration } } changes over hours and overnight, oven calibrated to 50 degrees, gelatin } } capsules totally filled and capped, in the oven since Friday. } } Help -- What's going wrong?? What to do now ?? } } Linda Fox } } lfox1-at-wpo.it.luc.edu } } I saw the post on LR white and noticed the no OsO4 part of the protocol. } I'm having problems with some LR white just now and I'm wondering if its } because the tissue (brown fat in this case) was Osmicated. The block seems } hard enough except where the tissue is and around there its like rubbery } butter. Interestingly enough the non osmicated blocks seem fine if maybe a } little too hard. Any ideas out there? } } Rod Nicholls } Head Technician } EM Unit } University of Ottawa } }
Although established as a tool in materials science and physics, scanning probe microscopy (SPM) is at the beginning of its application in biomaterials science. On 2 April 1998 the first workshop entitled "Scanning Probe Microscopy in Biomaterials Science, Dentistry and Medicine" was held in Bristol, UK. What was planned to be a small workshop evolved to be an international conference with high calibre delegates and speakers from all over the world. Encouraged by the success of the meeting and supported by international academics and industrial researchers we are organising a 2nd conference.
Since this first conference in 1998 more researchers have applied atomic force microscopy and related SPM methods in biomaterials science. Therefore a definitive need for a broad scientific exchange between researchers involved in these studies exists. This is the purpose of The 2nd International Conference on Scanning Probe Microscopy in Biomaterials Science, which will be hosted by the University of Bristol and Veeco Instruments Limited.
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Deadlines and dates
1 September 1999: early registration starts 1 January 2000: registration starts 1 April 2000 Deadline for abstract submission 1 June 2000 registration closes =96 late registration (at an increased fee rate) possible until the date of the conference.
---------------------- Manuela Finke University of Bristol Department of Oral and Dental Science Dental Materials and Biomaterials Group Lower Maudlin Street Bristol BS1 2LY tel:0117/9284537 fax:0117/9284780 e-mail:M.Finke-at-bris.ac.uk
} I'm wondering if vapor osmication will work on dry sections. I have } sections stained for immuno gold and the protocol calls for grids to be } floating on pbs during the vapor fix. I'm wondering if I need to have them } wet and then have to wash them or can they be done dry and thus save the } sections from the rigors of washing and pbs floating. Thanks in advance. } } Rod Nicholls } Head Technician } EM Unit } University of Ottawa
I have successfully both reosmicated sections that had the Os chemically removed and osmicated sections of various polymer blends using the vapor phase. I mount the dry grids on a glass microscope slide place it in a glass Petri dish and add a ml or so of 2% OsO4/H2O to the dish bottom (not touching the grids) put the top on and let it sit for some period of time (I usually figure an hour or so, but have never tested the rate). Do this under an approved fume hood.
Good luck
John Heckman TEM Specialist MSU Center for Electron Optics
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I have used OsO4 vapour fixation, both on floating and dry grids and have found that adequate results are achieved either way. If your resin is slightly hydrophilic (e.g. LR White), then, floating the grids on aqueous OsO4 will produce better results without causing widespread deposition, of Os around the gold particles.
} } } I'm wondering if vapor osmication will work on dry sections. I have } sections stained for immuno gold and the protocol calls for grids to be } floating on pbs during the vapor fix. I'm wondering if I need to have them } wet and then have to wash them or can they be done dry and thus save the } sections from the rigors of washing and pbs floating. Thanks in advance. } } Rod Nicholls } Head Technician } EM Unit } University of Ottawa } } }
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'd be willing to bet that the omission of accelerator is the cause of your trouble. Usually, I used temperatures of about 60C for 48-72 hr ,in the absence of oxygen (important), to achieve polymerisation.. Best results for immunohistochemistry were achieved using UV polymerisation at 0-4C for 24-48 hrs, though others have attributed loss of antigenicity to the UV. I have been able to achieve polymerisation of the resin using osmicated tissue, and my understanding is that heat generated locally around the Os deposits interferes with antigenicity rather than with resin polymerisation. For IHC work, aqueous uranyl acetate produces some increase in contrast, particularly of cell membranes, which are tough to see in the absence of osmium. Some respondents have mentioned sectioning problems. LR White is tough to section! I concur with others who advised REALLY infiltrating the tissue. Dehydration to 100% ethanol seemed to help, followed by LR White:EtOH (1:1) 1-3 hrs (depending on block size) LRW:EtOH (3:1) 1-3hrs Then, three changes of 100% LRW over 24 hrs at 4C before embedding. Having said all this, the rather poor appearance of non-osmicated , LR White-embedded sections led us to move away from IHC on embedded sections, to using pre- embeddibg methods whenever possible (more often than the literature might lead one to believe). } Thanks again!! The problem may be that there was no catalyst added } to the resin...(one should always ask the obvious questions when } novices are about ... Thank you) I re-read all the proceedures and } nowhere does it state to add the benz. perox. to the resin. I assumed } that the b.p. was needed for the cold cure method, where excess heat might } be generated, so I did not add any. I reinfiltrated the tissues with } another bolttle of resin, from EMS, catalyst added by manufacturer, and } returned them to the oven. We will see!! Thanks for all the tips. Many } called for higher temps. for polymerization. I am wondering if this will } hinder immuno work? I will be persuing this technique for one scientist } over the next few months and would appreciate all the tips, tricks and } protocols you can spare. Thanks, Linda Fox lfox1-at-wpo.it.luc.edu Fax: } 1-708-216-3676 }
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
For many years we have taught the practical aspects of SEM & TEM to post-graduate students to enable them to undertake their EM projects. We are now considering the possibility of instituting a 12 week module (1 lecture + 1 prac per week) on electron microscopy (history, development, optics, vacuum, sample prep etc) We'll probably only just touch on EDX in this particular module. The students are likely to be 3rd year Science (including Geology)/Health Science undergrads most of whom have come from a technically disadvantaged background. I'm sure there are many out there who have/are successfully teaching such a module. I would like to make contact with you to get information on how you structured your course.
Thanks in advance
Mike Gregory Professor Mike Gregory Electron Microscope Unit, University of Durban-Westville Private Bag X54001 Durban, Natal, South Africa 4000
rod, i have used osmium vapour on dry lrwhite immuno-gold labbeled sections of plant tissue prior to uranyl and lead citrate staining. 30 mins vapour exposeure gave me good staining of the cell wall. vapour exposeure was carried out in chamber designed by owen& vesely vol.20/6 proceedings rms november 1985.
for preparing support films on grids we use Pioloform instead of Formvar because it is said that Pioloform support films are little more stable. When we use Grids mesh size 200 we have no problems. Now, for grids mesh size 100 our procedure does not work. The film always breaks. Does anybody know any tricks for preparing support films for grids with large mesh size?
-- Dr. Anne Heller, AG Elektronenmikroskopie, Institut f=FCr Botanik (210), Universit=E4t Hohenheim, Garbenstra=DFe 30 D-70593 Stuttgart
I thought I was alone in the tattered sections on grids department.
My algal tissue is generally pretty holey too, even after extended infilration times (up to 2 weeks). I put it down to the material being difficult, but maybe it is the resin. Ultrastructure is excellent in the bits that remain intact, and the blocks section pretty well. They were dehydrated to 100% alcohol, polymerised at 50C, and an immuno label is attatching to tissue components. I have a feeling the problem is tissue related, but how long infiltration in resin is required I cannot guess.
So no help really, but the more information available the more chance we have of somebody coming up with an idea.
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
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"Can't leave biological tissues in glutaraldehyde for long periods; Immunological and cytochemical reactions are killed by osmium; Must use glutaradehyde after labeling reactions on sections."
You are all under arrest for spreading widespread panic without producing = supporting documentation or direct personal experiences.
Paul Webster, Ph.D. House Ear Insititute 2100 West Third Street Los Angeles, CA 90057 213 273 8026 pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
by alumni.Princeton.EDU (8.9.1/8.9.1) with ESMTP id NAA28743 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 22 Sep 1999 13:10:26 -0400 (EDT) Sender: rick-at-alumni.princeton.edu Message-ID: {37E90F94.45D82069-at-alumni.princeton.edu}
Mike Ingram wrote:
} 1. Keep the repaired 30 mm detector at 142 eV... } 2. ... replace the crystal pack until it meets the 136 eV... } 3. Go to a 10 mm detector...
It depends on how you most often use the detector. For most applications, count rate (solid angle) is more valuable than resolution because you get more counts for less beam current, avoiding all the bad effects of high current (specimen damage, nuisance of changing apertures, worse contamination, and so on). So I would not recommend dropping back to a 10-sq-mm detector unless resolution is your primary criterion.
It also depends on how much light element work you do. Because the noise and energy-dependent terms for resolution add in quadrature, a 6 eV loss at 5.9 keV turns into a 10 or 11 eV loss at O Ka, which is also a more significant fraction of the peak width at O. If you do a lot of oxygen directly rather than by stoichiometry, for example, then maybe a 10 sq-mm detector is a better choice. But if you need resolution and paid for a premium detector, don't settle for a 136 eV 10-square. It should get down to 133 or 134 anyway.
Although this has always been a "magic number" for selling EDS, unless you do a lot of light-element work or trace analyses, a few eV one way or another isn't all that critical. How you feel about getting what you paid for is a different question.
After moaning about the poor ultrastructure for immuno achieved with LR White (a known downside of all acrylics) we switched to LR Gold (cold-UV). The ultrastructure was immensely better than LR White, although it did not approach that of epon embedments. When "White" or "Gold" capsules come out of the oven or out of the freeze box, keep the capsules closed and immediately place in a small vaccuum jar. Oxygen does seep through the gelatin when the capsules are left on the bench and thus will make it impossible to polymerize the blocks further a week later when one decides that the blocks are too soft. We put LR Gold capsules from the freezer box directly into a -20 freezer. This way there seems to be no adverse activity between the resin and the air, and we can at a later date decide what we want to do with the blocks before sectioning. I have used LR White extensively with osmium (oven cured). Obviously this was not an immuno project. At any rate, if you have difficulty with adequate ultrastructure (no osmication) for immuno, try the LR Gold. Have fun! Bye, Hildy
P.S. Do not try to embed chocolate in an acrylic. It is an ultra-mess!
Responding to the message of {37E8E69C.26A51222-at-uni-hohenheim.de} from Anne Heller {heller-at-Uni-Hohenheim.DE} : } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues, } } for preparing support films on grids we use Pioloform instead of Formvar } because it is said that Pioloform support films are little more stable. } When we use Grids mesh size 200 we have no problems. Now, for grids mesh } size 100 our procedure does not work. The film always breaks. Does } anybody know any tricks for preparing support films for grids with large } mesh size? } } } -- } Dr. Anne Heller, AG Elektronenmikroskopie, } Institut f=FCr Botanik (210), Universit=E4t Hohenheim, } Garbenstra=DFe 30 } D-70593 Stuttgart }
Dr. Heller,
We use 0.5% formvar in ethylene dichloride to coat 1 x 2 mm slot grids. We do not carbon coat the formvar. These are the only grids we use in our laboratory. For 15 years we have used a JEOL 100CX microscope to look at these grids. The formvar is very stable and never breaks in the beam. I think older microscopes have "hotter" beams that may cause problems with the formvar.
John Basgen
John M. Basgen Department of Pediatrics University of Minnesota Box 491 420 Delaware Street SE Minneapolis, MN 55455 Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-tc.umn.edu
Greetings, Paul Webster called our attention to certain fixed ideas that showed up recently: } } "Can't leave biological tissues in glutaraldehyde for long periods; } Immunological and cytochemical reactions are killed by osmium; } Must use glutaradehyde after labeling reactions on sections."
I just wanted to add that LR white is not the only resin for doing immunocytochemistry. If LR white is giving you fits, try something different. Someone working with me just got nice immuno-gold results with material embedded in butyl-methyl-methacrylate. OK, this resin is not as stable in the TEM beam as LR white, but it is (usually) child's play to work with. A reasonable trade off? Your call.
These are all current Olympus metallurgical objectives, if you are interested in any or all the objectives please send us a reasonable offer, these are like new (they where purchased new for a project and never used).
The objectives are;
MSPLAN 2.5x / 0.07 Olympus Part Number 1-LM 510
ULWD MSPLAN 20x / 0.40 Olympus Part Number 1-LM 546
ULWD MSPLAN 50x / 0.55 Olympus Part Number 1-LM 556
ULWD MSPLAN 80x / 0.75 Olympus Part Number 1-LM 594
Hmm... what's the jurisdiction for this infraction? Will they tried in the MSA tribunal high court? If found guilty, will they be sentenced to a career of working with only RCA model scopes?
We need to talk to Nestor about how to file an injunction and restraining order to protect the rights of hapless biologicals floating in osmium...
Respectfully submitted by the court clerk,
Laura :)
} "Can't leave biological tissues in glutaraldehyde for long periods; } Immunological and cytochemical reactions are killed by osmium; } Must use glutaradehyde after labeling reactions on sections." } } You are all under arrest for spreading widespread panic without producing } supporting documentation or direct personal experiences. } } Paul Webster, Ph.D. } House Ear Insititute } 2100 West Third Street } Los Angeles, CA 90057 } 213 273 8026 } pwebster-at-hei.org } http://www.hei.org/htm/aemi.htm } ********************************************** Laura S. Rhoads US Distributor- AM-Toffeln
L. S. Rhoads P. O. Box 554 Johnson City, NY 13790-0554
I have never had any problems getting the resin to polymerize in gel caps even at 50 degree, but I have found that I have less sticky tops where the top of the capsule still traps air by going to just 52 to 55 degree and the labeling will still work.
I also get samples that cut difficult, seeming to disintegrate in the boat or look broken up in the scope. I have looked back at the protocol and suspect poor fixation. The higher I can get that glut the better cutting I get.
How many people have tried that Unicryl? I had questionable results, and heard the same from some one else.
Thanks for all the advice on tissue storage. There seemed to be a split on storing in fix vs bufer, but most buffer people stored after osmication.
My next problem is how to keep the vibatome sections flat while processing. These are 50 microns. previous work has been 100 to 150 microns and did not have the problem. These thinner ones tend to become cup shaped and or wavy. Some one here suggested sandwich them between glass slides or plastic sheets. 1) how to keep plastic from floating 2) will enough fluids get undernieth (sect. are 3 to 5 mm square) Any other ideas out there? Hopefully simple ones...... I have 39 more of the darn things. Thanks again
If you would like to stay with Pioloform, the only solution is to increase the plastic concentration in your solution. Sometimes film becomes unstable because of water traces in the plastic's solution. Therefore, I prefer to store the plastic itself in the vacuum dessicator permanently and I am using HPLC-grade freshened from time to time organic solvents. I am surprised to know that Pioloform is more stable under the beam than Formvar. Personally, I prefer Formvar on 150 mesh grids. The disadvantage of using Formvar film that it is less transparent to the e-beam. If this is a problem, I am using "old friend" - Parlodion stabilized by carbon. Generally speaking it is a very good idea to stabilize plastic film by carbon. It takes a bit more time to prepare but the quality will completely compensate time you spent for it. Using hex-grids instead square-hole grids may help to stabilize film too.
Good luck to you.
} Date: Wed, 22 Sep 1999 13:05:05 -0500 } From: John Basgen {basgen-at-maroon.tc.umn.edu} } Subject: Re: support film } To: Microscopy-at-sparc5.microscopy.com } X-Mailer: POPmail 2.3b8 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Sergey Ryazantsev UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
You might try a 0.5% solution of Formvar in (very dry) ethylene dichloride to coat your 200 mesh grids. Floating on water, the film should be silver to barely silver/gold. We use this color film for coating 1 x 2 mm slot grids and rarely have a problem with broken films. Sometimes we use a slightly gray/silver film with some breakage. We do not coat with carbon. For slot grids we need to pick up the film w/grids using a support made from aluminum (or other firm support...not parafilm) or the film sags, contacts the parafilm and sticks to it. We coat glass slides with the formvar solution and also dry them in a dry nitrogen gas atmosphere (using a "glove bag") to avoid problems with atmospheric humidity causing holes in the films. When dry they are removed from the bag in the fume cabinet (noxious ethylene dichloride fumes) and floated at ambient room humidity.
Hope this helps,
Doug ---------------------- Douglas R. Keene Shriners Hospital Microscopy Facility 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
I asked a question about expectant mothers and the teratogenic hazards in the typical EM facility a couple of weeks ago and it is now time to summarize. There were about 20 answers on and off list. They were all reasoned, informed opinions that cited literature, personal experience, the law, safety organizations, and common sense. They broke out like this (if you don't mind my offbeat characterizations):
A) Check the MSDS, and gloves, a fume hood, maybe a radiation monitor and common sense are all that are needed. Go for it. 5 males and 1 female.
B) I've done it, all turned out fine - just be careful 3 females
C) I've done it, but stayed clear of the chemicals - all was OK, be careful 3 females
D) I've done it, had problems, who knows if the lab was the culprit? 2 females
E) Just don't do it! 1 male
F) Watch for law suits - discrimination if you try to exclude her, liability if you don't. 2 females 1 male
These answers probably don't add up properly, because some people had more than one answer. I have purposely not quoted any answer because some were off list and were very personal and revealing. The general idea is quite sufficient, I think, and I thank all who took the time and emotional courage to write to help another person make a big decision.
That decision (made by the student with your help and that of our on campus safety/hazards people) was not to take the class this term. Our student is an undergrad who was/is working as a tech in another lab and who would greatly extend her usefulness if she were able to do the EM for her Prof., but whose job is not on the line and whose grad work is not yet a concern, so perhaps her decision was a little easier. If so, thankfully.
Again, thanks to Nestor for all the trench work and grief, and all the List Citizens for taking time and sharing your cumulative years of experience and judgement. I am in awe of this group!! Regards, Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (602)-965-3210 Fax - (602)-965-6899
Linda, The problems we've encountered when using LR White without the catalyst involved using: a) LRW - 6 months past date of purchase b) 70% ETOH made from an opened bottle of supposedly 100% (Remedy: pretest your 70-85% ETOH with LRW--if it turns cloudy---there's too much water in the alcohol. Also, we embedd in gelatin capsules topped with several drops of mineral oil to seal the acrylic during polymerization. I have not been happy with any embedding we've done using the catalyst. Rosemary
} } } } Robb, } } Per SAE Aerospace Recommended Practice ARP 598B(12/86), "13.2 } Competence - } Individual laboratories, with experienced counters, should be able to count with } a 10% mean deviation thus proving operator and optical accuracies." } } Bob Holthausen } Principal Microscopist } Pall Corporation } } } } } } "Rob.Calabro/-at-astrapharmaceuticals.com" on 09/21/99 05:50:26 PM } } To: Microscopy-at-Sparc5.Microscopy.Com } cc: (bcc: Bob Holthausen/SLSNY/Pall/US) } Subject: Counting Repeatability using LM Techniques } } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all, } } Does any information or publication exist that describes or defines the } "typical" variability or range that may be observed between analysts for } particle counting techniques using LM? Thanks. } } Robb Calabro } Pharmaceutical Scientist } AstraZeneca Pharmaceutical } Westborough, MA USA } } } } }
Tobias got a point. There are other embedding media that the LR's. I am surprised that the hydrophilic Lowicryls has not been mentioned yet. They are excellent for immunowork in the EM, and if you are worried about using fixatives, try a brief fixation in freshly made paraformaldehyde, fast dehydration in DMP, and then drop the temperature to -50 C and embed and cure at this temperature in Lowicryl. It works for some types of plant material. However, some people are allergic to Lowicryls so take care.
Bo
Tobias Baskin wrote:
} Greetings, } Paul Webster called our attention to certain fixed ideas that } showed up recently: } } } } "Can't leave biological tissues in glutaraldehyde for long periods; } } Immunological and cytochemical reactions are killed by osmium; } } Must use glutaradehyde after labeling reactions on sections." } } I just wanted to add that LR white is not the only resin for } doing immunocytochemistry. If LR white is giving you fits, try } something different. Someone working with me just got nice } immuno-gold results with material embedded in } butyl-methyl-methacrylate. OK, this resin is not as stable in the TEM } beam as LR white, but it is (usually) child's play to work with. A } reasonable trade off? Your call.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi all, } } Does any information or publication exist that describes or defines the } "typical" variability or range that may be observed between analysts for } particle counting techniques using LM? Thanks. } } Robb Calabro } Pharmaceutical Scientist } AstraZeneca Pharmaceutical } Westborough, MA USA
Robb,
I don't think there is "typical" variability. Each parameter measured or counted will have its own variability depending on inherent biological variation among animals, variability due to sampling, and measuring errors.
There are methods for partitioning these different levels of variability so that you can determine how much precision is necessary at the different levels of sampling and measuring error so you can detect a defined amount of difference between experimental groups. The book "Unbiased Stereology" by C.V. Howard and M.G. Reed has a good summary of these methods. At $32 this book may be the best buy in the scientific literature.
Remember, when counting particles, they are 3-dimensional. If you are making sections (2-D samples) through the 3-D particles and making counts on the sections you are not counting the particles but are counting 2-D profiles of the 3-D particles. The number of 2-D profiles is not directly related to the number of particles in the 3-D tissue. This book will also describe the method for counting particles in 3-D tissue from 2-D samples.
I assume you are talking about preparing slices for TEM. For brain slices, I rarely have problem of curling slices until dehydration. I found it helps by starting with a lower alcohol concentration (e.g. from 30%,).
Before the final polymerization step, I keep them flat by sandwitching the slices between two Aclar sheets. Then the sandwitch is further put between two glass slides with a paper clip to keep them flat. You may have few crackes for those curled slices. but there are always more slices for EM sectioning anyway.
No support document but some personal experiences ;-).
Cheers,
Xuejun
On Wed, 22 Sep 1999 rlvaughn-at-unmc.edu-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Thanks for all the advice on tissue storage. There seemed to be a split on } storing in fix vs bufer, but most buffer people stored after osmication. } } My next problem is how to keep the vibatome sections flat while processing. } These are 50 microns. previous work has been 100 to 150 microns and did not } have the problem. These thinner ones tend to become cup shaped and or } wavy. Some one here suggested sandwich them between glass slides or } plastic sheets. } 1) how to keep plastic from floating } 2) will enough fluids get undernieth (sect. are 3 to 5 mm square) } Any other ideas out there? Hopefully simple ones...... I have 39 more of } the darn things. } Thanks again } } Rick Vaughn } } }
*********************************************************************** Xuejun Sun, Ph.D. Dept. of Experimental Oncology Cross Cancer Institute 11560 University Ave. Edmonton Alberta T6G 1Z2, Canada
Exclusion of Oxygen is critical to the polymerisation of LR White. Most people appear to be using Gelatin capsules for this, but no-one has mentioned flat embedding. We have used a flat embedding technique for years and have got very reliable results from LR white. The samples, after infiltration, are placed in small plastic molds and overfilled with LR White. A sheet of glass/metal is then placed carefully on top of the molds and held in place by a heavy weight. The resin is then polymerised under vacuum. The polymerised samples can be cut out and mounted on perspex rods for sectioning. The only problem is finding the unosmicated samples after embedding, but the polymerisation was reliable every time. The resin itself is quite brittle to section, but a certain amount of the section damage can be caused before thin sectioning. I found that trimming the block face with a glass knife using water in the boat minimised the early damage and left the sample in good condition for sectioning with a diamond knife. Early fracture damage can penetrate into the block face and fall out during thin sectioning. The catalyst being used by a number of people is a cold setting catalyst. These catalysts tend to generate heat themselves and if used with a 50 degree oven will probably raise the sample temperature above 50 degrees.
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: Rosemary Walsh [SMTP:rw9-at-psu.edu] Sent: Thursday, September 23, 1999 1:16 AM To: microscopy-at-sparc5.microscopy.com
Linda, The problems we've encountered when using LR White without the catalyst involved using: a) LRW - 6 months past date of purchase b) 70% ETOH made from an opened bottle of supposedly 100% (Remedy: pretest your 70-85% ETOH with LRW--if it turns cloudy---there's too much water in the alcohol. Also, we embedd in gelatin capsules topped with several drops of mineral oil to seal the acrylic during polymerization. I have not been happy with any embedding we've done using the catalyst. Rosemary
Does anyone have a good electropolish solution and procedure for Ti-6-4. Any help would be greatly appreciated. Thanks. ______________________________________________ Roberto Garcia Senior Analyst, Metallography NC State University / Analytical Instrumentation Facility Campus Box 7531 Room 318 EGRC 1010 Main Campus Dr. Raleigh, NC 27695-7531 (919) 515-8628 (919) 515-6965 Fax rgarcia-at-unity.ncsu.edu http://spm.aif.ncsu.edu/aif ____________________________________________
A colleague asks: If anyone has got the answer to this problem, please help! Particularly with Heteroderidea subjects, putting them onto the stub is not a problem until slushing in liquid nitrogen. I usually find only three or four nematodes out of say 20 - 50 remaining, which is a big loss and means a lot more work. The endoparasitic females are OK but J2 and males are a very large problem. J2 stage 500um long. Chris Smith, Plant Path EM Unit, IACR-Rothamsted, UK.
This may be off of the normal topics for this group, but I thought I would ask anyway.
I am looking for Laboratory Information Management software for our applications lab at Rodel. Our applications lab has a variety of process and metrology equipment used in its day to day operation. Our products are polishing materials for the semiconductor industry. The LIMS should be able to:
1. Schedule and track experiments 2. Provide status of work in progress 3. Provide summary of completed work 4. Provide access to raw data 5. Collect data from process and metrology tools for analysis including raw data, images and enable data analysis. 6. Provide look-up and summaries from past work via keyword search.
Are any of you working with such a LIMS system and if so is it working for you? Any recommendations? Michael Ingram Rodel, Inc. 451 Bellevue RD Newark, DE 19713 Phone: 302.366.0500, ext. 2545 Fax: 302.455.1124
I have just been asked to come up with a system for automatically replenishing the liquid nitrogen in the anticontamination device for our TEM (it's a Philips CM200). We often want to run the 'scope overnight (in cryo mode, in an automated search for viruses), and we have already lengthened the time between replenishments by fitting a larger nitrogen flask onto the instrument. Does an automatic liq. nitrogen filler already exist? Please let me know ...
Thanks
Scott Robinson Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 N Mathews Ave., Urbana IL 61801
The worms are probably being ripped off the stub by the turbulent boiling of the liquid nitrogen. We have experienced the same problem with e.g. fungal conidial chains, the sporangiophores emerging nude from the liquid nitrogen. A fix for this is to cryofix by placing the stub on a metal (copper, aluminium) block just covered by a pool of liquid nitrogen in a polystyrene box. Adjust the liquid level so that the surface of the stub is not submerged. Cryofixation will be slower than by immersion in nitrogen slush, but this is of little importance if you want surface information only. If you regard fast cryofixation as essential, then the same principle can be applied to worms mounted on small pieces of thin metal sheet or foil of much lower mass, em grids even. After cryofixation, these may be mounted on the stub using a clamping screw or spring clip etc.
Hope this helps Chris Jeffree } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A colleague asks: If anyone has got the answer to this problem, please } help! } Particularly with Heteroderidea subjects, putting them onto the stub } is not a } problem until slushing in liquid nitrogen. I usually find only three } or four } nematodes out of say 20 - 50 remaining, which is a big loss and means } a lot } more work. The endoparasitic females are OK but J2 and males are a } very large } problem. J2 stage 500um long. } Chris Smith, Plant Path EM Unit, IACR-Rothamsted, UK. } } }
===================================================================== 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 =====================================================================
Fair go Paul. That "Dogma Police" header is a trifle emotive and not justified? The first two examples of "Dogma" given may be mine, but they are out of context. I had written that storing tissues in GA affects high resolution imaging. I gave reasons. Are you asserting that excessive fixation in GA does not affect high resolution imaging???
I had written {Immunological and cytochemical reactions are killed by osmium} , but the "killed" was in quotation marks, because I had learned about 30 years ago, that most such cellular activities cease . I do not believe that cells or osmium have changed in this regard. I have no reason to change my mind - or do you have any evidence to the contrary. I guess about half of all subscribers would like to know about that. To use your words "were is your supporting documentation" Please Paul, cite some publications were full osmium fixation was used prior to immunolabeling. 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 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, September 23, 1999 2:17 AM, Paul Webster [SMTP:pwebster-at-mailhouse.hei.org] wrote:
} "Can't leave biological tissues in glutaraldehyde for long periods; } Immunological and cytochemical reactions are killed by osmium; } Must use glutaradehyde after labeling reactions on sections." } } You are all under arrest for spreading widespread panic without producing } supporting documentation or direct personal experiences. } } Paul Webster, Ph.D. } House Ear Insititute } 2100 West Third Street } Los Angeles, CA 90057 } 213 273 8026 } pwebster-at-hei.org } http://www.hei.org/htm/aemi.htm } }
On Thursday, September 23, 1999 7:17 AM, Laura Rhoads [SMTP:laura-at-lsrhoads.com] wrote: } Hmm... what's the jurisdiction for this infraction? Will they tried in the } MSA tribunal high court? If found guilty, will they be sentenced to a } career of working with only RCA model scopes? } Laura, this is funnier than you thought. I did a two year stint on the RCA TEM that had graced the World Expo in Montreol (about 1969?), that scope later went to Prince Edward Island. A little while ago that RCA scope was mentioned on this listserver in "hushed tones"; I guess it has historical value. I would not want to offend anybody, but I've been in purgatory for two years already. Circa 1970 that instrument was the worst money could buy in the Western World. 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 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
I have a recipe for jet "sectioning" for Ti-6A1-4V which is as follows:
13% HCl 87% methanol
Temperature -60 degrees C Jet Height: 3.17mm Flow Rate: Medium Volts: 60 Current: 60mA
This was done with a South Bay Technology Single Vertical Jet Electropolisher. If you are using an instrument from another manufacture= r (twin jet system) then the reference to "jet height" would not be relevan= t. Other parameters may need to be adjusted, but the basic recipe should work.
As for the term jet "sectioning", it refers to the removal of a uniformly=
thick layer of metal by electropolishing for a given amount of time in an=
appropriate electrolyte using a specified set of parameters. The amount = of metal removed is determined by the length of time that current is allowed=
to flow through the cell while other parameters are held constant. =
Sectioning is used in a variety of applications where information is soug= ht on the structure of a material at some distance below the surface. These=
include studies of radiation damage, oxidation and ion implantation of doping elements. This differs from jet "thinning" in that with jet "thinning" you are removing material in such a way as to produce a concavity or dimple - the apex of which eventually perforates the specime= n. With that in mind, the flow rate and jet height may need to be adjusted even on our single jet system if you are trying to dimple the sample as opposed to section it.
More detailed discussion on the jet sectioning technique, as well as jet polishing in general, can be found in "Polishing Methods for Metallic and=
Ceramic Transmission Electron Microscope Specimens" by B.J. Kestel. (most=
of what I just said came out of that report) This report is available fro= m South Bay Technology at no charge upon request. I know this is a more detailed answer than you probably wanted, but I felt the additional information would be useful. I hope it helps.
} } } } } 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 "Roberto Garcia" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Hello Everyone,
Does anyone have a good electropolish solution and procedure for Ti-6-4. Any help would be greatly appreciated. Thanks. ______________________________________________ Roberto Garcia Senior Analyst, Metallography NC State University / Analytical Instrumentation Facility Campus Box 7531 Room 318 EGRC 1010 Main Campus Dr. Raleigh, NC 27695-7531 (919) 515-8628 (919) 515-6965 Fax rgarcia-at-unity.ncsu.edu http://spm.aif.ncsu.edu/aif ____________________________________________ {
} I have just been asked to come up with a system for automatically } replenishing the liquid nitrogen in the anticontamination device for our } TEM (it's a Philips CM200). We often want to run the 'scope overnight (in } cryo mode, in an automated search for viruses), and we have already } lengthened the time between replenishments by fitting a larger nitrogen } flask onto the instrument. Does an automatic liq. nitrogen filler already } exist? Please let me know ...
Dear Scott, We have a set of CryoMiser automatic LN fillers for our HVEM. On the plus side, they keep the LN levels full for traps which are located in high-radiation areas; however, on the minus side, the solenoid valves which they actuate fail now and again. Usually they fail so that the coil opens, thus the valve is closed, but occasionally they fail so that the coil
is fused in the energized state, whereupon the LN continues to pour into whatever device it's attached to. Once we had a coil fail that way, heat up, and ignite the foam insulation on the LN line, starting a small fire in the scope room. As Murphy would have it, this occurred on a Friday evening of a long weekend. We now only use these units during the day when someone is here to rectify any disasters. Yours, Bill Tivol
I have a couple of questions regarding the determination of glass fiber length distribution in a polymer matrix.
We normally determine the glass fiber length using a number of steps that involve:
1) ashing of the polymer matrix 2) mounting of the fibers on a glass slide with a suitable liquid medium to help disperse the fibers 3) OM and image analysis to determine fiber length distribution
Our concern is that the actual fiber length might change during the sample preparation.
So, does anyone know of an alternate method to determine the glass fiber length distribution in the polymer matrix and does anyone have information on the errors involved in this kind of procedure ?
Also, does any one know of [glass fiber] standards that one can use to assess the accuracy of the whole procedure.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE: Dogma Police Dear Jim, Thanks for your reply. I had hoped for more hostile and numerous = responses but my mailbox is only full of support messages. I didn't mean = the message to be a personal attack, rather a way of provoking an open = discussion, something that is often lacking on this forum. =
My message was prompted mostly by the thread concerning publishing = information on-line, where the main objection was that there was = insufficient review process to make it credible. These comments followed = by the solid statments I quoted (admittedly out of context) clearly = illustrated the point of what poor refereeing can result in.
I agree that for teaching purposes, a little dogma is not a bad thing and = have been guilty of it myself. However, to see you quote text in your web = site as a support for your claims made me think we should start discussing = the finer details of what we are doing here.
Firstly, some references where osmium fixation has been used for = immunolabeling. Ottersen's group in Olso (1984 J. Comp Neurol 229:374-= 392; 1986 Med. Biol. 64:147-158; 1990 Prog Brain Res 83:99-114) applied = antibodies to brain sections that had been fixed in 2.5% glutaraldehyde, 1%= formaldehyde, treated with 1% osmium tetroxide, dehydrated in ethanol and = propylene oxide and embedded in epoxy resin! I am sure I could find more = without too much effort if you want them (I remember what I read, not = where I read it).
I see Tamara Howard has provided additional additional references and the = comments that there are few very hard and fast rules in EM specimen = preparation. I agree totally with this. It is important that we keep = attempting to try out new things so that we can push the bounderies. = Setting limits during the early stages of instruction stops growth into = areas we can never imagine.
Jim, your other comment, that long fixation affects resolution is also not = documented fact. I do remember a paper, which I will look for, where = extraction in aldehyde fixative was dependant upon the buffer being used, = not the aldehyde. What is clearly documented is that the subsequent = processing steps (post-fixation, dehydration and infiltration) affect the = amount of extraction and thus reduce resolution. =
For most morphology applications, contrast is preferred over resolution = anyway. Witness the problem of getting immunocytochemical results = published over the last 10 years. Even here on the listserver, there is = always an ongoing discussion of how to get better contrast in the LR or = Lowicryl resins. When we all finally accept that contrast equals loss of = antigen, we may eventually enjoy the esthetics of low contrast, high = information images, and learn to leave the high contrast pictures in the = morphology books.
If high resolution is required then there are better methods than those = involving aldehyde fixation and epoxy resin embedding. For the poster of = the original question I would say, leave the tissues in glutaraldehyde for = as long as is convenient, any loss of structure will go unnoticed. = However, as with all advice, I reccommend you take it with a pinch of salt = and try it out for yourself.
Finally, the issue of fixing immunolabeled sections with 1% glutaraldehyde = came up and there were a variety of answers. The one I took mild = exception to was from Jan Leunissen. He is a great teacher and I respect = him and his work at many levels. In his message, he correctly pointed out = that the conditions of high salt and low pH are used to elute antibodies = from antigens. Therefore 1% glutaraldehyde must be used to crosslink = antibodies to sections and protein A-gold to antibodies. It all sounds = good in theory but where is the proof? I and my colleagues have been = applying antibodies and colloidal gold to sections for years without that = final crosslinking step. Why do we still get specific label? Who knows. = What we do know is that we have compared the effect of adding this = fixation step and decided that it didn't improve labeling efficiency. We = therefore made an informed decision to leave it out of the protocol. = Putting unneccessary steps into a protocol only makes life more difficult = for people working in busy labs. =
My advice for anyone just starting in EM, is to take what you are told = with healthy skepticism and question anything you feel may be unreasonable.= If there is a reason for doing something your teacher should know that = reason. If they don't know why something should be done, try leaving it = out to see if it makes a difference. To apply methods on the faith that = they work is not sufficient anymore. We need informed reasons for = applying methods. We are in the good times for EM and we have to shed our = artist/magician image. The closer we get to being scientists, the more = seriously we will be accepted by the science community who are at present = looking for our skills.
For the record, I also take exception to the methods that involve the use = of bodily fluids. If we can't work out how to do something without nose = or face grease we really deserve to be left behind.
Thanks Jim, for taking the bait and allowing the start of what I hope will = be an informative discussion on credibility.
Regards,
Paul Webster.
=
jim wrote: } Fair go Paul. That "Dogma Police" header is a trifle emotive and not = justified? = } The first two examples of "Dogma" given may be mine, but they are out of = } context. } I had written that storing tissues in GA affects high resolution imaging. = I = } gave reasons. } Are you asserting that excessive fixation in GA does not affect high = resolution = } imaging??? } } I had written {Immunological and cytochemical reactions are killed by = osmium} , = } but the "killed" was in quotation marks, because I had learned about 30 = years = } ago, that most such cellular activities cease . I do not believe that = cells or = } osmium have changed in this regard. I have no reason to change my mind - = or do = } you have any evidence to the contrary. I guess about half of all = subscribers = } would like to know about that. To use your words "were is your supporting = } documentation" Please Paul, cite some publications were full osmium = fixation = } was used prior to immunolabeling. } 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 } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com } } On Thursday, September 23, 1999 2:17 AM, Paul Webster = } [SMTP:pwebster-at-mailhouse.hei.org] wrote: } } } "Can't leave biological tissues in glutaraldehyde for long periods; } } Immunological and cytochemical reactions are killed by osmium; } } Must use glutaradehyde after labeling reactions on sections." } } } } You are all under arrest for spreading widespread panic without = producing } } supporting documentation or direct personal experiences. } } } } Paul Webster, Ph.D. } } House Ear Insititute } } 2100 West Third Street } } Los Angeles, CA 90057 } } 213 273 8026 } } pwebster-at-hei.org } } http://www.hei.org/htm/aemi.htm } } } } } = } } } RFC822 header } ----------------------------------- } } Received: from ultra.ultra.net.au [203.20.237.5] by mailhouse.hei.org = with = } ESMTP } (SMTPD32-4.07) id A00314BB021E; Thu, 23 Sep 1999 07:58:11 PST } Received: from 150 (p098.supa2-tsv.ultra.net.au [202.80.71.98]) } by ultra.ultra.net.au (8.9.3/8.9.3) with SMTP id BAA13065; } Fri, 24 Sep 1999 01:02:04 +1000 (EST) } Received: by localhost with Microsoft MAPI; Fri, 24 Sep 1999 01:02:01 += 1000 } Message-ID: {01BF0628.68BC69A0.jim-at-proscitech.com.au} } From: jim {jim-at-proscitech.com.au} } Reply-To: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } To: "'Paul Webster'" {pwebster-at-mailhouse.hei.org} , } "'MSA listserver submission'" } {Microscopy-at-sparc5.microscopy.com} } Subject: RE: Dogma Police } Date: Fri, 24 Sep 1999 00:50:03 +1000 } Return-Receipt-To: jim {jim-at-proscitech.com.au} } Organization: ProSciTech } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } MIME-Version: 1.0 } Content-Type: text/plain; charset=3D"us-ascii" } Content-Transfer-Encoding: 7bit } X-UIDL: 234843928 } Status: U }
We are interested in possibly purchasing a used Reichert Ultracut E with FC4E cryo unit. If you have an Ultracut E and/or FC4E to sell (in good working order) please contact me directly.
Fred A. Hayes The DOW Chemical Co Analytical Sciences, Microscopy 1897 bldg., E78 Midland, Michigan 48667 517-638-2203 517-638-6443 fax
Craig, S. and Goodchild, D.J. (1984). Periodate-acid treatment of sections permits on-grid immunogold localization of pea seed vicilin in ER and Golgi. Protoplasma 122, 35-44.
Chris
} I had written {Immunological and cytochemical reactions are killed by osmium} , } but the "killed" was in quotation marks, because I had learned about 30 years } ago, that most such cellular activities cease . I do not believe that cells or } osmium have changed in this regard. I have no reason to change my mind - or do } you have any evidence to the contrary. I guess about half of all subscribers } would like to know about that. To use your words "were is your supporting } documentation" Please Paul, cite some publications were full osmium fixation } was used prior to immunolabeling. } 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 } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com } } On Thursday, September 23, 1999 2:17 AM, Paul Webster } [SMTP:pwebster-at-mailhouse.hei.org] wrote: } } } "Can't leave biological tissues in glutaraldehyde for long periods; } } Immunological and cytochemical reactions are killed by osmium; } } Must use glutaradehyde after labeling reactions on sections." } } } } You are all under arrest for spreading widespread panic without producing } } supporting documentation or direct personal experiences. } } } } Paul Webster, Ph.D. } } House Ear Insititute } } 2100 West Third Street } } Los Angeles, CA 90057 } } 213 273 8026 } } pwebster-at-hei.org } } http://www.hei.org/htm/aemi.htm } } } } } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
{ { { { Does anyone have a good electropolish solution and procedure for Ti-6-4. Any help would be greatly appreciated. Thanks. ______________________________________________ Roberto Garcia} } } }
Roberto,
Try the following:
660ml Metahanol 400 ml 2-Butoxy-ethanol 66ml Perchloric Acid
} Thanks for all the advice on tissue storage. There seemed to be a split } on storing in fix vs bufer, but most buffer people stored after } osmication. } } My next problem is how to keep the vibatome sections flat while } processing. These are 50 microns. previous work has been 100 to 150 } microns and did not have the problem. These thinner ones tend to become } cup shaped and or wavy. Some one here suggested sandwich them between } glass slides or plastic sheets. 1) how to keep plastic from floating 2) } will enough fluids get undernieth (sect. are 3 to 5 mm square) Any other } ideas out there? Hopefully simple ones...... I have 39 more of the darn } things. Thanks again } } Rick Vaughn } Dear Rick,
I worked almost exclusively with Vibratome sections (rat CNS, immunoEM, 15-30 um) for ~10 yrs. We tried many things to preserve shape prior to embedding. The best system consisted of CARE and a few tricks (which follow). These assume that the sections will be osmicated before embedding. If not, things are a little easier. Once the sections are ready to dehydrate, I'd make a tool out of a 9" Pasteur pipet heated over a Bunsen burner. The tip is pulled thin and at an angle slightly greater than 90 degrees (~100-120 degrees) to the pipet. It is then curved around to form a circle (polygon) of a size adequate to support the sections without allowing them to fold. It takes a few tries to get a good one. This is used to transfer the sections into OsO4, which is the most critical step. You need to take great care to insure that the sections are flattened within seconds after they hit the osmium (perferably before), because they will rapidly be "frozen" into shape by this fixative. Since they rapidly become brittle, folded sections have to be broken to unfold them and wrinkles are made permanent, which leads to uneveness or breakage during embedding.I did the osmication in wide mouthed scintillation vials that allowed complete flattening of each coronal rat brain section. Because time is spent adjusting the sections one at a time in an open container of 1-2% OsO4, this MUST be done in a fume hood with the sash at the height that insures no efflux of the vapour. Use gloves, too. Rinsing was done in the same vial. After the fixation period (1-3 hrs for us), the sections will be very brittle. Sections thicker than 25um are fairly tough, but some care should be taken in their handling. Thinner sections are tricky to handle and may still fold. Some breakage is hard to avoid, but with care they won't look like jigsaw puzzles after embedding. Washes in ascending concentrations of dehydrant were done in separate containers wide enough to ensure that the sections were deposited flat. I found this to work better than doing all the changes in the same container with the } 25um sections. The thinner sections could not be handled so much. I found that I could maintain better control of the final shape of the thicker sections as I transferred them, but there was still a risk of breaking them. As dehydration progresses, they become even more brittle, so one has to be very gentle near the end. All of the same issues apply to non- osmicated sections, but the risk of folding increases, while the risk of breakage decreases. Infiltration was done in epoxy-acetone rather than in one of the more traditional transitional solvents. This permitted infiltration to take place in plastic weighing boats, where the sections could be gently teased as flat as possible against the bottom of the boat with the Pasteur pipet tool. These sections were transferred to weigh boats containing pure warmed resin. which was changed once. The resin was warmed after 1 hr. infiltration and sections were then GENTLY (they break very easily at this point due to resin surface tension- which is why warmed (40-60C) resin is used- and the weight of the resin on the tissue) transferred on to a pre-treated glass slide (1% dichlorodimethyl silane in benzene, then dried for at least 4 hr). I included accelerator (DMP-30) in the warmed resin, so infiltration times had to be short ( {90 min) or polymerization would start. The sections were oriented, excess resin was removed and another similarly-treated slide placed on top and gently pressed down. The slide sandwiches were then placed in the embedding oven. This basic method worked for LR White emdedding protocols, which is easier to work with, due to its lower viscosity. If your sections are about 50 um thick, things should be quite simple. Here's my $ 0.02 about section storage: After 4% paraformaldehyde fix, sections were still fine for immunoEM after several months in PBS with 0.02% Na azide. Up to 2 months in PBS+4% para with Na azide, immunoreactivity and ultrastructure were OK, but sections had to be VERY thoroughly rinsed. After osmication, sections were OK in buffer for a few weeks. After months, the membranes and other osmophilic structures had a granular appearance. If you store in osmium for more than a day or two, you can heavily stain golgi apparatus and membranes will be dark, but granular. Proteins are extracted too, I have read. Haven't tried long- term storage in glutaraldehyde. I hope some of the above will be helpful. 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
Chris, Although this is not a fast freeze, we've been happy with nematodes on thermanox coverslips (cut in half) to fit the slide holder for the Oxford cryotrans system. I directly place it on the cold stage in the specimen prep chamber and freeze it there as opposed to plunging into the liquid nitrogen slush. To minimize excess moisture I transfer with a fine brush. Other options: coat coverslips with a thin layer of carbon/OCT or poly-lysine. Rosemary
I was wondering if anyone has any advice for working with GACH (glutaraldehyde/carbohydrazide) resin. I've been looking at water-miscible embedding media and it came up as a possibility, but the carbohydrazide refuses to completely dissolve (at 150mg carbohydrazide per mL of 50% glut), even when added in separate stages accompanied by rapid stirring. I spun down the tube anyway and just used the supernatant but it spontaneously polymerized at room temperature (in some cases before the infiltration was complete) into opaque light tan blocks. Is that normal? I would also be interested to hear what experience anyone has in working with melamine resin (not the commercial Nanoplast preparation, but actually made from formaldehyde and melamine). Thanks in advance for any suggestions anyone can offer.
Jason Davis M.Sc. student Department of Medical Biophysics University of Toronto
For studying polymer blends by using SEM, only information on surface of a sample can be obtained. Because the surface can not always be the plane of the center of each dispersed phase particle, the statistic result of dispersed phase particle size attained from SEM micrograph may be less than the true value. This error can be corrected by assuming the particles are spheres with equal diameter d and randomly distributed: d=8r/pi where r is the observed average radius. However, I can not find a equation for the ellipse particles. Does anyone has knowledge on this problem?
Thank you in advance for your help.
Zeng Jijun Visiting Scholar of Toray
Home Address: Nakatogari 734-2-5 Nagaizumi-cho, Sunto-gun Shizuoka, 411-0942 Japan
For studying polymer blends by using SEM, only information on surface of a sample can be obtained. Because the surface can not always be the plane of the center of each dispersed phase particle, the statistic result of dispersed phase particle size attained from SEM micrograph may be less than the true value. This error can be corrected by assuming the particles are spheres with equal diameter d and randomly distributed: d=8r/pi where r is the observed average radius. However, I can not find a equation for the ellipse particles. Does anyone has knowledge on this problem?
Thank you in advance for your help.
Dr. Zeng Jijun Visiting Scholar of Toray
Home Address: Nakatogari 734-2-5 Nagaizumi-cho, Sunto-gun Shizuoka, 411-0942 Japan
With regard to upholding the "honor" of RCA, the first firm to commercialize a TEM in the world (so far as I know): ========================================== If found guilty, will they be sentenced to a } career of working with only RCA model scopes? =========================================== Of the several column instruments (SEMs and TEMs) under our roof, one of them is an RCA EMU4-B, it was manufactured in 1969 and delivered to us, after serving as a "demo" instrument at a university for a year, in 1972. That TEM has been in faithful operation ever since. Over the years, it has been used every day by a variety of different users in a busy analytical services and quality inspection laboratory. It is the "back up" when our more modern TEM goes down....yes, it does require service, but not any more so than a newer TEM and apparently a lot less than some newer TEMs. The total costs for service on the RCA in most years have been less than the service costs associated with our more modern systems.
While it might not have been for everyone, at the time of its manufacture it was essentially all solid state and it took roughly another ten years for other manufacturers to achieve that level of solid state design (and reliability). Technicians in those days who worked by day using one of the imported TEMs and by night, for SPI, on our RCA always marveled at how it just kept going, and going and going.....
For us, it has been a real "workhorse" for inspecting filmed grids, high volume running of stained rubber modified polymer samples, polymer coatings for defects, and latex particles for size distribution, and yes, even a lot of life science tissue type work. There are obviously many things it can not do that a more modern TEM could do, but for those who are doing the "routine" kinds of TEM work as just indicated, and at not especially high resolution requirements, contrary to what seemed to have been getting suggested, at least we are quite proud to be the owner of this RCA TEM. Admittedly, this "senior citizen" of a TEM will be retired before too long, however, we are very pleased with the level of performance it has given us over the years.
To this day, at least some of us believe it was a real loss to the EM community when RCA decided to drop out of this business. Some day, perhaps , some graduate student, for a thesis project, will research the reasons why RCA suffered its untimely demise, possibly leading to lessons for future generations.
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 ######################## ============================================
} From: "chris.smith" {chris.smith-at-bbsrc.ac.uk} Priority: normal
In regard to Chuck Garber's comments on the RCA EMU-4, I have one important addition: CONTRAST! In my 20+ years of TEM use, I've never seen a scope match the contrast of the EMU-4 for biological specimens. I remember the rhythm of the clatter and bang of the film carriers and the skill necessary just to get a grid into the column. I guess the only bad thing I remember about that scope was the amount of heat generated by the freestanding electronic components. Years ago, lack of space forced us to decide to keep only one of our two scopes. It broke my heart when I found out they hauled it (and the identical unit at the VA Hospital) off to the landfill. I was able to preserve a few mementos, a gun cap, the filament centering jig and some filaments. A sad story.
Bob Santoianni Emory University Hospital Atlanta, GA
I have a polymer membrane which contains an ester R group. The surface of the membrane has been acid hydrolyzed to convert the ester to an alcohol. I need to measure the depth to which the hydrolysis reaction has occurred and get some qualitative sense for the concentration gradient of OH groups within the modified layer. ATR-FTIR of the surface indicates that the hydrolysis reaction worked, however, the SE image of the cross section shows no change in morphology.
Can someone think of a way to selectively label the OH groups to provide some contrast in the SEM? How about RuO4 (Macromolecules 1983, 16, 589-598)? I expect that the modified layer could be as thin as a few hundred nanometers. Would this preclude the use of EDS or BSE on the labeled cross section? If so, would labeling the OH groups effect the contrast in the SE signal?
Thanks in advance,
Brian Reid UTD Chemistry breid-at-utdallas.edu 972-883-2709
Yes, they still exist. I have come across two and they are up for grabs (you pay shipping!). So before they hit the dumpster, give me a call. Lynne Garone Polaroid Corp. 1265 Main St. W4-1D Waltham, Ma. 02451 GaroneL-at-Polaroid.com 781 386-1446
Chuck, I think that a Reference work of the history of RCA in this endeavor would be a welcome addition to all of our bookshelves. I wonder how we all might go about getting something moving on this? Any other input from the rest of the list folk?
As a resource, we have most of the back issues of the various electronics magazines, which would provide some of the engineering background, and if I could find what box they are in RCA also had an engineering publication as well.
Ed Sharpe archivist for SMECC
} Subj: RCA TEMs (good old days) } Date: 9/24/99 1:52:24 AM US Mountain Standard Time } From: cgarber-at-2spi.com (Garber, Charles A.) } To: Microscopy-at-Sparc5.Microscopy.Com (MICROSCOPY BB) } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } With regard to upholding the "honor" of RCA, the first firm to commercialize } a TEM in the world (so far as I know): } ========================================== } If found guilty, will they be sentenced to a } } career of working with only RCA model scopes? } =========================================== } Of the several column instruments (SEMs and TEMs) under our roof, one of } them is an RCA EMU4-B, it was manufactured in 1969 and delivered to us, } after serving as a "demo" instrument at a university for a year, in 1972. } That TEM has been in faithful operation ever since. Over the years, it has } been used every day by a variety of different users in a busy analytical } services and quality inspection laboratory. It is the "back up" when our } more modern TEM goes down....yes, it does require service, but not any more } so than a newer TEM and apparently a lot less than some newer TEMs. The } total costs for service on the RCA in most years have been less than the } service costs associated with our more modern systems. } } While it might not have been for everyone, at the time of its manufacture it } was essentially all solid state and it took roughly another ten years for } other manufacturers to achieve that level of solid state design (and } reliability). Technicians in those days who worked by day using one of the } imported TEMs and by night, for SPI, on our RCA always marveled at how it } just kept going, and going and going..... } } For us, it has been a real "workhorse" for inspecting filmed grids, high } volume running of stained rubber modified polymer samples, polymer coatings } for defects, and latex particles for size distribution, and yes, even a lot } of life science tissue type work. There are obviously many things it can } not do that a more modern TEM could do, but for those who are doing the } "routine" kinds of TEM work as just indicated, and at not especially high } resolution requirements, contrary to what seemed to have been getting } suggested, at least we are quite proud to be the owner of this RCA TEM. } Admittedly, this "senior citizen" of a TEM will be retired before too long, } however, we are very pleased with the level of performance it has given us } over the years. } } To this day, at least some of us believe it was a real loss to the EM } community when RCA decided to drop out of this business. Some day, perhaps } , some graduate student, for a thesis project, will research the reasons why } RCA suffered its untimely demise, possibly leading to lessons for future } generations. } } C
I can't take credit for the following technique for getting flat, osmicated sections, so I'll share it with you, asking the indulgence of whoever was the inventor. The sections come out flat every time.
Working in a fume hood, half-fill a small Petri dish with buffer and wet several millipore filters in it. Choose a filter size that accomodates your sections. Transfer one unosmicated section to the dish and manuever it over a filter. Carefully lift the filter out of the buffer so the section lies flat. Transfer to another, larger empty Petri dish. Apply osmium (I use 1% in PB or PBS) dropwise, 3 or 4 drops will usually do, and allow section to sit in osmium a few minutes. Replenish the drops to keep the section moist, if needed. While one section is fixing, I use the time to line up the next section. After fixing on the filter, transfer filter/section to another dish of buffer. Gently brush or wiggle the section off the filter into the buffer. The sections are stiffer than unosmicated sections but not yet brittle. I transfer them to a small vial of fresh osmium with a paintbrush, but you could use a spatula of some sort if you're worried they might crack. The sections flatten back out in the osmium and I leave them to fix further for another hour. After fixing and rinsing, the sections go on to a typical dehydration protocol. They mostly retain their flatness, with occasionally a little ruffling along the edges, throughout the dehydration and infiltration steps.
Good luck. Hope this helps.
Missy
Eleanor Josephson, DVM, PhD University of Connecticut Health Center Department of Anatomy MC-3405 263 Farmington Ave. Farmington, CT 06030-3405 Ph.(860)679-2463 Fax (860)679-1274 ejosephs-at-neuron.uchc.edu
I agree with you. I don't think that building dogmas about this or that is a good scientific way of thinking. There are so many variables involved in the different technical procedures that predictions may fail. Of course, we have to begin from some point, but sometimes I've found myself doing things that people had said me that will never work and - surprise (even to me) - it worked!
Coming back to glutaraldehyde, it is obvious that the results may vary with the tissue, the buffer or the species that we are studying. In my experience the "dogmas" that work for mammals don't work for reptiles or fishes. Try to fix caiman digestive system with less than 5% glutaraldehyde and you will see what I' m trying to explain. However I had excellent results in fishes with 1% glutaraldehyde. I also embedded materials that were kept for 6 months in 2.5% glutaraldehyde in phosphate or cacodilate buffer with excellent results even at high magnification.
The good procedure to your material is the one that makes your work easier and still gives good results.
Sincerely
Francisco Javier Hernandez-Blazquez IARC-WHO Unit of Multistage Carcinogenesis 150 Cours Albert Thomas 69372 - Lyon France Telephone: (33) 472738536 Fax: (33)472738442
fhernandez-at-iarc.fr
Paul Webster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Reply to: RE: Dogma Police } Dear Jim, } Thanks for your reply. I had hoped for more hostile and numerous responses but my mailbox is only full of support messages. I didn't mean the message to be a personal attack, rather a way of provoking an open discussion, something that is often lacking on this forum. } My message was prompted mostly by the thread concerning publishing information on-line, where the main objection was that there was insufficient review process to make it credible. These comments followed by the solid statments I quoted (admittedly out of context) clearly illustrated the point of what poor refereeing can result in. } } I agree that for teaching purposes, a little dogma is not a bad thing and have been guilty of it myself. However, to see you quote text in your web site as a support for your claims made me think we should start discussing the finer details of what we are doing here. } } Firstly, some references where osmium fixation has been used for immunolabeling. Ottersen's group in Olso (1984 J. Comp Neurol 229:374-392; 1986 Med. Biol. 64:147-158; 1990 Prog Brain Res 83:99-114) applied antibodies to brain sections that had been fixed in 2.5% glutaraldehyde, 1% formaldehyde, treated with 1% osmium tetroxide, dehydrated in ethanol and propylene oxide and embedded in epoxy resin! I am sure I could find more without too much effort if you want them (I remember what I read, not where I read it). } } I see Tamara Howard has provided additional additional references and the comments that there are few very hard and fast rules in EM specimen preparation. I agree totally with this. It is important that we keep attempting to try out new things so that we can push the bounderies. Setting limits during the early stages of instruction stops growth into areas we can never imagine. } } Jim, your other comment, that long fixation affects resolution is also not documented fact. I do remember a paper, which I will look for, where extraction in aldehyde fixative was dependant upon the buffer being used, not the aldehyde. What is clearly documented is that the subsequent processing steps (post-fixation, dehydration and infiltration) affect the amount of extraction and thus reduce resolution. } For most morphology applications, contrast is preferred over resolution anyway. Witness the problem of getting immunocytochemical results published over the last 10 years. Even here on the listserver, there is always an ongoing discussion of how to get better contrast in the LR or Lowicryl resins. When we all finally accept that contrast equals loss of antigen, we may eventually enjoy the esthetics of low contrast, high information images, and learn to leave the high contrast pictures in the morphology books. } } If high resolution is required then there are better methods than those involving aldehyde fixation and epoxy resin embedding. For the poster of the original question I would say, leave the tissues in glutaraldehyde for as long as is convenient, any loss of structure will go unnoticed. However, as with all advice, I reccommend you take it with a pinch of salt and try it out for yourself. } } Finally, the issue of fixing immunolabeled sections with 1% glutaraldehyde came up and there were a variety of answers. The one I took mild exception to was from Jan Leunissen. He is a great teacher and I respect him and his work at many levels. In his message, he correctly pointed out that the conditions of high salt and low pH are used to elute antibodies from antigens. Therefore 1% glutaraldehyde must be used to crosslink antibodies to sections and protein A-gold to antibodies. It all sounds good in theory but where is the proof? I and my colleagues have been applying antibodies and colloidal gold to sections for years without that final crosslinking step. Why do we still get specific label? Who knows. What we do know is that we have compared the effect of adding this fixation step and decided that it didn't improve labeling efficiency. We therefore made an informed decision to leave it out of the protocol. Putting unneccessary steps into a protocol only makes life more difficult for people working in busy labs. } My advice for anyone just starting in EM, is to take what you are told with healthy skepticism and question anything you feel may be unreasonable. If there is a reason for doing something your teacher should know that reason. If they don't know why something should be done, try leaving it out to see if it makes a difference. To apply methods on the faith that they work is not sufficient anymore. We need informed reasons for applying methods. We are in the good times for EM and we have to shed our artist/magician image. The closer we get to being scientists, the more seriously we will be accepted by the science community who are at present looking for our skills. } } For the record, I also take exception to the methods that involve the use of bodily fluids. If we can't work out how to do something without nose or face grease we really deserve to be left behind. } } Thanks Jim, for taking the bait and allowing the start of what I hope will be an informative discussion on credibility. } } Regards, } } Paul Webster. } } } } jim wrote: } } Fair go Paul. That "Dogma Police" header is a trifle emotive and not justified? } The first two examples of "Dogma" given may be mine, but they are out of } context. } } I had written that storing tissues in GA affects high resolution imaging. I } gave reasons. } } Are you asserting that excessive fixation in GA does not affect high resolution } imaging??? } } } } I had written {Immunological and cytochemical reactions are killed by osmium} , } but the "killed" was in quotation marks, because I had learned about 30 years } ago, that most such cellular activities cease . I do not believe that cells or } osmium have changed in this regard. I have no reason to change my mind - or do } you have any evidence to the contrary. I guess about half of all subscribers } would like to know about that. To use your words "were is your supporting } documentation" Please Paul, cite some publications were full osmium fixation } was used prior to immunolabeling. } } 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 } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } } } } On Thursday, September 23, 1999 2:17 AM, Paul Webster } [SMTP:pwebster-at-mailhouse.hei.org] wrote: } } } } } "Can't leave biological tissues in glutaraldehyde for long periods; } } } Immunological and cytochemical reactions are killed by osmium; } } } Must use glutaradehyde after labeling reactions on sections." } } } } } } You are all under arrest for spreading widespread panic without producing } } } supporting documentation or direct personal experiences. } } } } } } Paul Webster, Ph.D. } } } House Ear Insititute } } } 2100 West Third Street } } } Los Angeles, CA 90057 } } } 213 273 8026 } } } pwebster-at-hei.org } } } http://www.hei.org/htm/aemi.htm } } } } } } } } } } } } } RFC822 header } } ----------------------------------- } } } } Received: from ultra.ultra.net.au [203.20.237.5] by mailhouse.hei.org with } ESMTP } } (SMTPD32-4.07) id A00314BB021E; Thu, 23 Sep 1999 07:58:11 PST } } Received: from 150 (p098.supa2-tsv.ultra.net.au [202.80.71.98]) } } by ultra.ultra.net.au (8.9.3/8.9.3) with SMTP id BAA13065; } } Fri, 24 Sep 1999 01:02:04 +1000 (EST) } } Received: by localhost with Microsoft MAPI; Fri, 24 Sep 1999 01:02:01 +1000 } } Message-ID: {01BF0628.68BC69A0.jim-at-proscitech.com.au} } } From: jim {jim-at-proscitech.com.au} } } Reply-To: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } } To: "'Paul Webster'" {pwebster-at-mailhouse.hei.org} , } } "'MSA listserver submission'" } } {Microscopy-at-sparc5.microscopy.com} } } Subject: RE: Dogma Police } } Date: Fri, 24 Sep 1999 00:50:03 +1000 } } Return-Receipt-To: jim {jim-at-proscitech.com.au} } } Organization: ProSciTech } } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } MIME-Version: 1.0 } } Content-Type: text/plain; charset="us-ascii" } } Content-Transfer-Encoding: 7bit } } X-UIDL: 234843928 } } Status: U } }
I would like to capture on video the analysis that I perform on the SEM. There is a video out from the SEM. What equipment is required? Can I take the signal directly from the CRT? I do not have any experience with this. Any help is much appreciated.
TIA
David Rose W.L. Gore & Associates 297 Blue Ball Road Elkton, MD 21921 410-506-2958
We have an LKB 7801A glass knifebreaker that may need servicing and or minor parts. Does anyone service this knifebreaker and does anyone know where I can buy minor replacement parts resulting from wear?
Fred A. Hayes The DOW Chemical Co Analytical Sciences, Microscopy 1897 bldg., E78 Midland, Michigan 48667 517-638-2203 517-638-6443 fax
If I take your assumptions (all spheres of identical diameter), you should find particles of all diameters up to a maximum diameter. That would be the diameter of the particles (there is no way of getting a cut that produces a 2D diameter larger than the 3D diameter for a sphere). Likewise you could look at the shape factor of elliptical particles. If you make the same assumptions (all particles identical), the largest (or smallest, depending on definition) shape factor should be the one you are looking for.
This is of course only true if:
a) you have enough particles to get a representative distribution, and b) your elliptical particles are rotationally symmetric around one of the axes.
If you are looking at the histogram of diameters (for the spherical particles), you might also be able to extract the diameter distribution. You would have to calculate what the measured histogram for a given diameter would look like (looks like you have done that), fold that with a distribution (for example a gaussian distribution) and then fit this function to the observed histogram. You might be able to extract the mean diameter and the width of the distribution that way.
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 ===================================
-----Original Message----- } From: Exchange Administrator Sent: Thursday, September 23, 1999 11:44 PM To: Michael Bode
Dear list members:
For studying polymer blends by using SEM, only information on surface of a sample can be obtained. Because the surface can not always be the plane of the center of each dispersed phase particle, the statistic result of dispersed phase particle size attained from SEM micrograph may be less than the true value. This error can be corrected by assuming the particles are spheres with equal diameter d and randomly distributed: d=8r/pi where r is the observed average radius. However, I can not find a equation for the ellipse particles. Does anyone has knowledge on this problem?
Thank you in advance for your help.
Dr. Zeng Jijun Visiting Scholar of Toray
Home Address: Nakatogari 734-2-5 Nagaizumi-cho, Sunto-gun Shizuoka, 411-0942 Japan
Lehigh University seeks an Electron Microscope Technician to perform technical duties in support of the Electron Microscopy Laboratory of the Materials Science and Engineering Department. Technician will be a member of a team responsible for the following: instruct students in the operation of microscopes and other equipment; maintain and repair instruments; oversee upkeep of the lab; support research professors and students; analyze samples; give tours and demonstrations; maintain a safe environment; and other assigned duties. Bachelors degree in physical science and/or 4+ years related work experience required. Candidates should be familiar with electron microscopes, mechanical and electronic equipment, vacuum systems, PC and/or MAC and EDS/WDS systems. Good communication and interpersonal skills are essential.
Lehigh University offers excellent benefits and vacation package. Interested candidates should forward resume to: Deanne Hoenscheid, Materials Science and Engineering, Lehigh University, 5 E. Packer Ave., Bethlehem, PA 18015. EOE. M/F/D/V.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } "Can't leave biological tissues in glutaraldehyde for long periods; } Immunological and cytochemical reactions are killed by osmium; } Must use glutaradehyde after labeling reactions on sections." } } You are all under arrest for spreading widespread panic without producing supporting documentation or direct personal experiences. } } Paul Webster, Ph.D. } House Ear Insititute } 2100 West Third Street } Los Angeles, CA 90057 } 213 273 8026 } pwebster-at-hei.org } http://www.hei.org/htm/aemi.htm } } } } Hi, Wonderful Reply! There are so many variations of these dogmas depending on the circumstances! Would take me a whole day and more to write all the exceptions to these absolutes (?).
On Wed, 22 Sep 1999 rlvaughn-at-unmc.edu-at-Sparc5.Microscopy.Com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have never had any problems getting the resin to polymerize in gel caps } even at 50 degree, but I have found that I have less sticky tops where the } top of the capsule still traps air by going to just 52 to 55 degree and } the labeling will still work. } } I also get samples that cut difficult, seeming to disintegrate in the boat } or look broken up in the scope. I have looked back at the protocol and } suspect poor fixation. The higher I can get that glut the better cutting I } get. } } How many people have tried that Unicryl? I had questionable results, and } heard the same from some one else. } } Rick Vaughn } } } Hi,
We have been using LR White and LR Gold. I investigated carefully the differences between these and Unicryl (numerous reports and papers), and I believe Unicryl to be advantageous. We will try it (exactly as it ought to be used by comparing directions from the company with reports in papers) in a "side by side" with LR Gold, which we favor for improved ultrastructure when compared to LR White - note - our protocol only. This last statement may not be viable for all procedures. Please pull all the papers you can find. Unicryl is likely to be superior to others.
Hildegard H. Crowley University of Denver {hcrowley-at-du.edu}
RCA must have had a long history of making EMs. I remember my high school days. My HS was across the street from Stanford Research Institute in Menlo Park, CA. SRI donated an RCA EMC-1 to the school. It did not function and no one knew what to do with it.
I thought it was the neatest thing I'd ever seen! It landed in our science club and I took about the task of making the SEM work. I did not know what a SEM really was and all the nuances associated with it. But I did know electronics. The SRI guys were experts at high vacuum and I learned a lot about the vacuum area from them.
The SEM used an oil diffusion pump of course. What would drive me crazy was that after a short period of time, the phosphor glass would get soaked in oil. I'd have to take the glass out, mix another slurry of phosphor and make a new screen. After about 2 months of this hassle, I asked the SRI guys about it. They said that I needed a cold trap. Oh. Were do I get one of those? "It already has it. Add LN2." Ahhh....what's LN2? Where do I get that? We did not have all that much adult supervision at the time and no one was going to let us have LN2.
Once I got the idea of what the cold trap was all about, I made a chiller by connecting the labs' refrigerator compressor cooling side around the diffusion pump where the LN2 would have gone. That worked pretty good. Not as good as LN2 but it lengthened the time of screen changes and made a huge difference in vacuum level.
when I got my first image on the screen it was really awesome. Well, not the image, just the fact that I got something to show up. Bugs and chromosomes (smashed of course) from Drosophila gonads looked cool.
HV was always a problem with the system. Lots of arcing. Well, in retrospect, anode-gun spacing and vacuum did not connect at the time.
After leaving the RCA SEM some 30 years ago, now I have my own SEM. It is quite different than the RCA of course--thankfully. And I know more now--and the expertise on this listserver is also indispensable. In the vein of Chuck's suggestion of why RCA left the EM market, I understand that Amray has also left the lab/research EM market after their buy-out by KLA-Tencor. From what I have heard, the market is too small to justify the cost of R&D. Amray made a bunch of 1000-series scopes and a ton of the 1800-series. The newest 3600 FESEM is really a great instrument. As far as I know, ISI/Topcon is not a big player in the SEM market any longer. Or maybe not a player at all. At least I can still get factory service for my Amray 1830T6. There are many independent service folks out there for both Amray and ISI systems. So that is good.
As these earlier makers depart the market, the options for replacement sources dwindles. But looking at the competition today, Hitachi, Jeol, Cambridge, Leo, etc. are all making very capable products. It seems to me that the rationale for being in or out of EM is the same as for any other field--money. Either it is profitable or not. Same for microelectronics, calculators (remember that highly competitive battle?), watches, VCRs, etc........ As the number of suppliers dwindles, hopefully the remaining few will survive since they will get new business and customers as a result of some other manufacturer exiting the marketplace. The danger of this is that without sufficient competition, innovation wanes. If no one is chasing you, why not just walk rather than run?
LKB sold their EM/Histology line to the company now known as Leica. They may still have some spare parts there.
800-248-0123 is the number.
-- Begin original message -- } } We have an LKB 7801A glass knifebreaker that may need servicing and or minor } parts. Does anyone service this knifebreaker and does anyone know where I } can buy minor replacement parts resulting from wear? } } Fred A. Hayes } The DOW Chemical Co } Analytical Sciences, Microscopy } 1897 bldg., E78 } Midland, Michigan 48667 } 517-638-2203 } 517-638-6443 fax } -- End original message -- 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)
If the signal that you get out of the SEM is a TV signal, you should be able to feed it directly into a VCR or a computer equipped with a frame grabber. There are several international standards for video (RS-170, NTSC, PAL, etc.). You need to make sure, whatever equipment you want to use is compatible with the standard used by your SEM. Also, call the manufacturer of the SEM to find out if the signal indeed follows those standards.
Once you get the signal into the computer, you can take easily take pictures and work with them (enhance them, annotate them, etc.)
One drawback of this technique is, that you are limited to the resolution provided by the TV standards (640x480 for US standards). And unless you have an internal frame store on the SEM, the images tend to be very noisy, as the beam must be scanned very rapidly to get the 25 or 30 frames per second required by the TV standards. For higher resolutions, you need either a passive or active SEM interface. Call me or send email if you need more information about those.
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: "drose-at-wlgore.com"-at-sparc5.microscopy.com[SMTP:"DROSE-at-WLGORE.COM"-at-SPARC5. MICROSCOPY.COM] } Sent: Friday, September 24, 1999 8:43:48 AM } To: microscopy-at-sparc5.microscopy.com } Subject: SEM - video recording } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear List,
I would like to capture on video the analysis that I perform on the SEM. There is a video out from the SEM. What equipment is required? Can I take the signal directly from the CRT? I do not have any experience with this. Any help is much appreciated.
TIA
David Rose W.L. Gore & Associates 297 Blue Ball Road Elkton, MD 21921 410-506-2958
Does anyone know where I could obtain a standard for TEM EDS analysis of the mineral sodalite (Na4Al3Si3O12Cl)? It would be preferable to have a bulk as opposed to powder sample.
I have tried Alpha Aesar, which doesn't carry it.
Many Thanks, Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
************************************************ Robert J. Derby New Mexico Institute of Technology Socorro, N.M. Phone - 505-835-5866 E-mail - rjderby-at-excite.com ************************************************ Greetings all, Dose anyone know where to get "Lippencott's Directory" (spelling might be off on Lippencott). I use to have a copy and now I can't find it and no luck on the net, nor interlibrary loan. It was a soft cover, out of Calf.,it listed all available antibodies and to all things, even "anti-gold" antibody, which was human if I remember. Any help would be a help.
Thanks
Robert Derby
________________________________________________________________ Get FREE voicemail, fax and email at http://voicemail.excite.com Talk online at http://voicechat.excite.com
Hayes, Fred (FA) wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We have an LKB 7801A glass knifebreaker that may need servicing and or minor } parts. Does anyone service this knifebreaker and does anyone know where I } can buy minor replacement parts resulting from wear? } } Fred A. Hayes } The DOW Chemical Co } Analytical Sciences, Microscopy } 1897 bldg., E78 } Midland, Michigan 48667 } 517-638-2203 } 517-638-6443 fax We recondition LKB Knife Makers, $175.00 plus parts and shipping. We sell pre-owned, reconditioned EM-, Histology-, Darkroom/Photo- and Lab Equipment.Please email your fax number for a list of available equipment. Services in EM and Histology since 1977. Microscopy Labs Box 338 61 West Street Red Bank, NJ 07701 732 747 6228 fax 732 758 9142
I am attempting to conduct dielectric function (constant) evaluations from Plasmon Loss EELS spectra acquired with our TEM using GIF. After doing a fast literature search I came up with two possible sources of information:
1) book by Egerton (1996 2nd edition) is supposed to contain recipe for calculations and a Fortran code used to transform EELS spectra into usable information (according to D.B. Williams and C.B. Carter). - I do not have a copy of the book, but could read it in a local University library or purchase it.
2) Program for a MAC described in 94 MSA meeting abstracts called Fantome which took EELS spectra in Gatan's format and gave back the dielectric functions. - I cannot find any information about it except for an old photocopy of the MSA abstract page.
Therefore, I am seeking an advice from the list.
Did anyone used the Fantome program and how could I obtain a copy of it, perhaps it evolved into something commercially available? Did anyone wrote your own code to do these analysis and would be willing to shear your experiences? Is there a Script/Plug-In for Gatan's software which would do just what I am seeking? How would one a) extract ASCII data from Gatan EL/P files, b) write code to evaluate them? Perhaps Mathematica could cajoled to use the Fortran code of Egerton as the core for calculations?
All comments regarding the above questions and the general calculations of dielectric functions would be greatly appreciated.
Respectfully yours.
Jerzy
***************************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Senior Materials Scientist 5204 E. Ben White Blvd. - MS 613 PCAL - Analytical TEM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 FAX: (512) 602-7470 jerzy,gazda-at-amd.com *****************************************************************
Please forgive the repeated mailing but my e-mail address had a typo in the first message. The correct e-mail address is: jerzy.gazda-at-amd.com . Original message is repeated below.
Dear Colleagues,
I am attempting to conduct dielectric function (constant) evaluations from Plasmon Loss EELS spectra acquired with our TEM using GIF. After doing a fast literature search I came up with two possible sources of information:
1) book by Egerton (1996 2nd edition) is supposed to contain recipe for calculations and a Fortran code used to transform EELS spectra into usable information (according to D.B. Williams and C.B. Carter). - I do not have a copy of the book, but could read it in a local University library or purchase it.
2) Program for a MAC described in 94 MSA meeting abstracts called Fantome which took EELS spectra in Gatan's format and gave back the dielectric functions. - I cannot find any information about it except for an old photocopy of the MSA abstract page.
Therefore, I am seeking an advice from the list.
Did anyone used the Fantome program and how could I obtain a copy of it, perhaps it evolved into something commercially available? Did anyone wrote your own code to do these analysis and would be willing to shear your experiences? Is there a Script/Plug-In for Gatan's software which would do just what I am seeking? How would one a) extract ASCII data from Gatan EL/P files, b) write code to evaluate them? Perhaps Mathematica could cajoled to use the Fortran code of Egerton as the core for calculations?
All comments regarding the above questions and the general calculations of dielectric functions would be greatly appreciated.
Respectfully yours.
Jerzy
***************************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Senior Materials Scientist 5204 E. Ben White Blvd. - MS 613 PCAL - Analytical TEM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 FAX: (512) 602-7470 jerzy.gazda-at-amd.com *****************************************************************
"Must use glutaraldehyde after labeling reactions on sections"
"You are all under arrest for spreading widespread panic without producing supporting documentation or direct personal experiences"
Although the first quote is not quite the way I put it, I assume the 'Must use...' statement refers to my posting on the listserver. The 'mild exception' Paul Webster takes with regard to my posting at least suggests this. I believe if you're arrested in the US you're entitled to make one phone call. Isn't that right? So I am using this privilege by using my dial-up account to transmit my comments to the Supreme Dogma Court. I hope my plea will get me out of jail again.
First, let me try to loosen up the jury a bit. I have always perceived the Listserver as a place for exchanging experiences in such a way that contributions are not necessarily declined or considered to be less valuable when not fully or explicitly documented, different from what at least is considered to be good practice in any scientific paper. I fully agree, that if such experiences are documented with experimental data, it gives the 'Listener' the possibility to evaluate the conclusions, at least to some extent. However, even with documented experiences, it is a matter of 'trust' whether the data supplied are perceived as being reliable. Years ago I was requested by referees to send in over 15 micrographs of control experiments for the evaluation of immunolabeling data in E.coli. The controls ideally should have no label. Easy to document, but reliable? Nevertheless, experiences may vary from group to group or from person to person: sometimes we may not know why, which certainly doesn't mean that these experiences are not real or valuable. The conclusions based on such experiences may in all sincerity be based on the wrong assumptions. Ladies and gentlemen, I don't think this is a problem at all, in comparing experiences finally the most reliable, or 'truthful' if you like,will emerge or survive. Documented by Darwin in a different context? The fun in sharing experiences seems to be in the end in preventing having to invent the wheel in each and every research facility over and over again and wasting preciou$ time (in more than one way). Exchanging experiences will at least help in thinking about experimental set-ups and sometimes prevent using too many unfruitful approaches. And after all, we still have our own responsibility to judge if we want to follow a certain idea or not.
And now for the hard part: Paul, Your Honour, my comments on the use of fixation after immunolabeling were not presented as a dogma, I wanted to give a reasonable explanation for its application. My message didn't state that fixation is a must. Documentation: direct personal experience as stated in the original posting. As you stated yourself this should be good enough, right? At least you use it for pleading your case. Our personal experiences often showed less signal when fixation was left out (but not with all immunoreagents) and this was also found by many of our workshop participants. Is this proof? I don't think it is, although I find it hard to find a different explanation. Anyway, I assume that many of the messages or comments on the List are based on personal experiences, which is just not mentioned explicitly all the time. I agree that taking up procedural steps which may complicate things should be avoided. But I don't agree that taking up a 5-10 minutes extra step in a procedure that may take from anywhere between an hour to several hours should make a difference to 'busy lab life', at least not when there is a good or likely reason to take it up.
So, Paul, either you set me free or you come in and join me in my prison cell. A nice weekend to all of you.
Cheers, Jan =========================== Jan Leunissen AURION http://www.aurion.nl Costerweg 5 6702 AA Wageningen phone: (31)-317-497676 fax: (31)-317-415955 You will find more tech info on our website.
I think you are looking for Linscott's Directory of Immunological and Biological Reagents. You can buy it either in hard copy or as a database for Windows or Mac; ordering information and contact addresses can be found at
A basic course on light microscopy which will cover the following topics:
Theory of microscopy Kohler Illumination Diffraction Theory Contrast Methods Polarized light Phase Contrast Interference Hoffman contrast Rheinberg Dark-field & oblique Illumination, etc.
The workshop will consist of four consecutive Saturdays of lectures and hands-on labs to cover the theoretical and practical aspects of microscopy.
The course instructors include:
Jan Hinsch of Leica, Inc.
Dennis O'Leary of Micro-Optical Methods
Mary McCann of McCann Imaging
John Reffner of Sensir Inc. and Current President of NYMS
Donald O'Leary, Curator and Awards Chairperson of NYMS
WHEN: October 2, 9,16, 23, 1999, from 10 A.M. to 4 P.M.
WHERE: 1244 McBride Avenue, West Paterson, NJ. Phone (973) 812-8377 (Free parking, accessible by public transportation, Information on car pools and transportation will be provided.)
COST: $225 for N.Y.M.S. members, $245 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: Beginners and experienced users who wish to learn more about the proper use of a microscope.
For Further Information and To Register contact Donald O'Leary.
I'm working on a project where we are trying to stretch a human hair under an electron microscope and create an animation of the process. We're planning on using some type of stepper motor driven linear actuator to do the work. I thought I'd check here to see if anyone could recommend a vendor for such motors/actuators. As we're new to this (SEM microscopy), any advice would also be much appreciated.
Most of the commercial phosphor plates I've seen for electron microscopy use P43 (blue light, fast decay). Can anyone suggest a source for Willemite, or P47, on similar glass plates or in powder?
I was wondering what computer programs people are using to drive microscopes and accessories (digital cameras, filter wheels, shutters) in the Macintosh world. I am familiar with NIH Image, but would like to hear about a few other programs. I should mention that I regularily use NIH Image, as well as some programs on the PC side. Are there any ones I should avoid, or are there any exceptionally good ones? Please feel free to reply to the list, or me directly.
cheers...
TIA
Eugene Krueger Research Technologist GI Research Mayo Clinic, Rochester (507) 284-0580 krueger.eugene-at-mayo.edu
Eugene, we use DigitalMicrograph to drive our electron microscope. We also use ti drive a separate stage control on this scope.
-- Jaap Brink, Ph.D., Biochemistry, One Baylor Plaza, Baylor College of Medicine, Rm. N420 Alkek Building, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Sun, 26 Sep 1999, Krueger, Eugene W. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all...... } } I was wondering what computer programs people are using to drive microscopes } and accessories (digital cameras, filter wheels, shutters) in the Macintosh } world. I am familiar with NIH Image, but would like to hear about a few } other programs. I should mention that I regularily use NIH Image, as well } as some programs on the PC side. Are there any ones I should avoid, or are } there any exceptionally good ones? Please feel free to reply to the list, } or me directly. } } cheers... } } TIA } } Eugene Krueger } Research Technologist } GI Research } Mayo Clinic, Rochester } (507) 284-0580 } krueger.eugene-at-mayo.edu } } }
Hi Paul & Listreaders: I accept that you cited the three examples of dogma taunting to stir some debate. At the very least you picked rather poor examples of "dogma". The third of your "dogmas" was submitted by Jan Leunissen and he presented an excellent plea showing his innocence and also that he was not properly quoted. Since Jan works for Aurion I assume that he knows a few things about immuno gold labeling, yet Paul had stated } without producing supporting documentation or direct personal experiences. {
Paul's quotation accusing me of dogma were incorrect too. } "Can't leave biological tissues in glutaraldehyde for long periods; { I actually had clearly qualified this as referring to HIGH RESOLUTION work AND I had given reason AND I have experience in that regard. Paul, have you looked at GA overfixed tissue at several hundred thousand times and compared that with properly fixed tissues? I stand by my original statement.
Paul's second misquote was: } Immunological and cytochemical reactions are killed by osmium; { A little, but vital omission were the quotation marks around the word killed. Afterall, we are talking about Immunological and cystochemical activities and these are not on-or-off, but diminish with various treatments. My original advise was like saying: "70% ethanol kills microorganisms". It does and most would be killed very rapidly, but there may be some, for instance yeast spores that could survive for some time in that high concentration of yeast "excreta". The statement "70% ethanol 'kills' . . . remains true. I did not say or imply that osmium "kills" all immune activities instantly and completely. I am not guilty of "dogma", which is a forcible statement or opinion stating as if unchallengable or authoritative assumptions rather than empirical observations.
It should be noted that a wrongly based forcible statement like Paul's "Dogma Police" actually suits that very definitions much better. I also see a couple of other problems with this kind of argument on the listserver. 1 It could deter some people from submitting freely their opinions. 2 It could lead to the frequent addition and confusing qualifiers and this would change the nature of this discussion forum.
Hands up, which of the other contributors to the topic did go back to my original message and read this and Paul's assertions critically? Anybody??? The replies come up with a few publications showing that at least some partially osmicated tissues, retained some antigens in some cases. Not many such publications though and we could safely assume that less than one in a hundred of such research projects used any osmium prior to labeling. This supports my case: Why would so many researchers publish such low contrast images IF normal EM preparation showed equally good activities? Paul agrees } When we all finally accept that contrast equals loss of antigen, we may eventually enjoy the esthetics of low contrast, high information images, and learn to leave the high contrast pictures in the morphology books. { } From that I infer that at least some of those "osmium before labeling" publications may have been "better" without osmium first. Furthermore, Paul here clearly states that higher contrast (e.g. osmication) equals lower antigenicity; in fact he is in agreement with my original advise. What as your point?
I admit that I too played a game. Paul used an emotive term to get people onsite. It worked, because we all detest "dogma". It would have been nice if critical thinking had prevailed. When I asked to provide prove that osmication does not "kill" immune activity {cite some publications were full osmium fixation was used prior to immunolabeling {, they came in like the tide.
You cannot prove that by showing that some activity is preserved in some case. What is required is a full comparison of remaining activity after different treatments in different tissues and different reactions. Safe your efforts, Paul and most of as know that: osmium "kills" immune activities.
Now, had somebody in reply to my original submission written in "your right Jim, but sometimes, after some osmium fixation there is enough remaining activity for successful labeling." I would have replied "Amen". 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 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, September 24, 1999 4:42 AM, Paul Webster [SMTP:pwebster-at-mailhouse.hei.org] 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 } -----------------------------------------------------------------------. } } } Reply to: RE: Dogma Police } Dear Jim, } Thanks for your reply. I had hoped for more hostile and numerous responses } but my mailbox is only full of support messages. I didn't mean the message } to be a personal attack, rather a way of provoking an open discussion, } something that is often lacking on this forum. } My message was prompted mostly by the thread concerning publishing information } on-line, where the main objection was that there was insufficient review } process to make it credible. These comments followed by the solid statments } I quoted (admittedly out of context) clearly illustrated the point of what } poor refereeing can result in. } } I agree that for teaching purposes, a little dogma is not a bad thing and have } been guilty of it myself. However, to see you quote text in your web site as } a support for your claims made me think we should start discussing the finer } details of what we are doing here. } } Firstly, some references where osmium fixation has been used for } immunolabeling. Ottersen's group in Olso (1984 J. Comp Neurol 229:374-392; } 1986 Med. Biol. 64:147-158; 1990 Prog Brain Res 83:99-114) applied antibodies } to brain sections that had been fixed in 2.5% glutaraldehyde, 1% } formaldehyde, treated with 1% osmium tetroxide, dehydrated in ethanol and } propylene oxide and embedded in epoxy resin! I am sure I could find more } without too much effort if you want them (I remember what I read, not where I } read it). } } I see Tamara Howard has provided additional additional references and the } comments that there are few very hard and fast rules in EM specimen } preparation. I agree totally with this. It is important that we keep } attempting to try out new things so that we can push the bounderies. Setting } limits during the early stages of instruction stops growth into areas we can } never imagine. } } Jim, your other comment, that long fixation affects resolution is also not } documented fact. I do remember a paper, which I will look for, where } extraction in aldehyde fixative was dependant upon the buffer being used, not } the aldehyde. What is clearly documented is that the subsequent processing } steps (post-fixation, dehydration and infiltration) affect the amount of } extraction and thus reduce resolution. } For most morphology applications, contrast is preferred over resolution } anyway. Witness the problem of getting immunocytochemical results published } over the last 10 years. Even here on the listserver, there is always an } ongoing discussion of how to get better contrast in the LR or Lowicryl } resins. When we all finally accept that contrast equals loss of antigen, we } may eventually enjoy the esthetics of low contrast, high information images, } and learn to leave the high contrast pictures in the morphology books. } } If high resolution is required then there are better methods than those } involving aldehyde fixation and epoxy resin embedding. For the poster of the } original question I would say, leave the tissues in glutaraldehyde for as } long as is convenient, any loss of structure will go unnoticed. However, as } with all advice, I reccommend you take it with a pinch of salt and try it out } for yourself. } } Finally, the issue of fixing immunolabeled sections with 1% glutaraldehyde } came up and there were a variety of answers. The one I took mild exception } to was from Jan Leunissen. He is a great teacher and I respect him and his } work at many levels. In his message, he correctly pointed out that the } conditions of high salt and low pH are used to elute antibodies from } antigens. Therefore 1% glutaraldehyde must be used to crosslink antibodies } to sections and protein A-gold to antibodies. It all sounds good in theory } but where is the proof? I and my colleagues have been applying antibodies } and colloidal gold to sections for years without that final crosslinking } step. Why do we still get specific label? Who knows. What we do know is } that we have compared the effect of adding this fixation step and decided } that it didn't improve labeling efficiency. We therefore made an informed } decision to leave it out of the protocol. Putting unneccessary steps into a } protocol only makes life more difficult for people wo } rking in busy labs. } My advice for anyone just starting in EM, is to take what you are told with } healthy skepticism and question anything you feel may be unreasonable. If } there is a reason for doing something your teacher should know that reason. } If they don't know why something should be done, try leaving it out to see } if it makes a difference. To apply methods on the faith that they work is } not sufficient anymore. We need informed reasons for applying methods. We } are in the good times for EM and we have to shed our artist/magician image. } The closer we get to being scientists, the more seriously we will be } accepted by the science community who are at present looking for our skills. } } For the record, I also take exception to the methods that involve the use of } bodily fluids. If we can't work out how to do something without nose or face } grease we really deserve to be left behind. } } Thanks Jim, for taking the bait and allowing the start of what I hope will be } an informative discussion on credibility. } } Regards, } } Paul Webster. } } } } } jim wrote: } } Fair go Paul. That "Dogma Police" header is a trifle emotive and not } } justified? } The first two examples of "Dogma" given may be mine, but they are } } out of } context. } } I had written that storing tissues in GA affects high resolution imaging. I } } } gave reasons. } } Are you asserting that excessive fixation in GA does not affect high } } resolution } imaging??? } } } } I had written {Immunological and cytochemical reactions are killed by } } osmium} , } but the "killed" was in quotation marks, because I had learned } } about 30 years } ago, that most such cellular activities cease . I do not } } believe that cells or } osmium have changed in this regard. I have no reason } } to change my mind - or do } you have any evidence to the contrary. I guess } } about half of all subscribers } would like to know about that. To use your } } words "were is your supporting } documentation" Please Paul, cite some } } publications were full osmium fixation } was used prior to immunolabeling. } } 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 } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } } } } On Thursday, September 23, 1999 2:17 AM, Paul Webster } } } [SMTP:pwebster-at-mailhouse.hei.org] wrote: } } } } } "Can't leave biological tissues in glutaraldehyde for long periods; } } } Immunological and cytochemical reactions are killed by osmium; } } } Must use glutaradehyde after labeling reactions on sections." } } } } } } You are all under arrest for spreading widespread panic without producing } } } supporting documentation or direct personal experiences. } } } } } } Paul Webster, Ph.D. } } } House Ear Insititute } } } 2100 West Third Street } } } Los Angeles, CA 90057 } } } 213 273 8026 } } } pwebster-at-hei.org } } } http://www.hei.org/htm/aemi.htm } } } } } } } } } } } } } RFC822 header } } ----------------------------------- } } } } Received: from ultra.ultra.net.au [203.20.237.5] by mailhouse.hei.org with } } } ESMTP } } (SMTPD32-4.07) id A00314BB021E; Thu, 23 Sep 1999 07:58:11 PST } } Received: from 150 (p098.supa2-tsv.ultra.net.au [202.80.71.98]) } } by ultra.ultra.net.au (8.9.3/8.9.3) with SMTP id BAA13065; } } Fri, 24 Sep 1999 01:02:04 +1000 (EST) } } Received: by localhost with Microsoft MAPI; Fri, 24 Sep 1999 01:02:01 +1000 } } Message-ID: {01BF0628.68BC69A0.jim-at-proscitech.com.au} } } From: jim {jim-at-proscitech.com.au} } } Reply-To: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } } To: "'Paul Webster'" {pwebster-at-mailhouse.hei.org} , } } "'MSA listserver submission'" } } {Microscopy-at-sparc5.microscopy.com} } } Subject: RE: Dogma Police } } Date: Fri, 24 Sep 1999 00:50:03 +1000 } } Return-Receipt-To: jim {jim-at-proscitech.com.au} } } Organization: ProSciTech } } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } MIME-Version: 1.0 } } Content-Type: text/plain; charset="us-ascii" } } Content-Transfer-Encoding: 7bit } } X-UIDL: 234843928 } } Status: U } } }
I'm a bit late in this discussion, have been trying to get on the list for a while -
I've been using Lowicryl K4M for nearly 10 years now, and only a few times have I had problems with sectioning. I can't recall ever having problems with the curing either (which have been the case with both Agar 100 and Araldite). I tried Unicryl for a while as well (both heat- and UV-polymerized), and my results were quite comparable with the Lowicryl ones. Has anyone compared Low/Uni with LR White or Gold?
Regards Margareta
Margareta Halin Dept. of Internal Medicine University Hospital Uppsala Sweden
We are having problems with the developing of SO163 films. We get streaks on the negatives and the results are highly inconsistent in terms of exposure/optical density. We don't have these problems when we use 4489, but we need the higher sensitivity of the SO163 for cryo work. Has anybody else experienced such problems?
I think at least some of these problems are related to the lack of temperature control of our developing solutions. Now, we may be able to keep the developer at the proper temperature by further cooling of the room with air-con. However, is the temperature of the tap water used for rinsing also very important? This is in Singapore, and our tap water is usually very warm, sometimes close to 30C.
If this is important, does anybody have experience with water cooling systems for this purpose? A recirculating cooling water bath could be used for cooling the developer, but wouldn't work for the rinse, for which we would probably need some kind of in-line cooler. (Maybe similar to that used in water drinking fountains?) I am concerned about the potentially very high expense of such systems.
Any advice would be greatly appreciated. Thanks,
Terje
------------------------------------ Dr. Terje Dokland Senior Scientist Institute of Molecular Agrobiology 1 Research Link The National University of Singapore Singapore 117604 Phone: 65-872 7405 Fax: 65-872 7007 E-mail: dokland-at-ima.org.sg http://lab.ima.org.sg/lab3052/terje ------------------------------------
Mr. Egerton placed all the source codes that are discussed in his book and some other helpful files and data on the following ftp-site
ftp://ftp.phys.ualberta.ca/public_html/eels/
Cheers,
Petra
At 19:54 24.09.99 -0500, you wrote:
*SNIP*
} I am attempting to conduct dielectric function (constant) evaluations from } Plasmon Loss EELS spectra acquired with our TEM using GIF. After doing a } fast literature search I came up with two possible sources of information: } } 1) book by Egerton (1996 2nd edition) is supposed to contain recipe for } calculations and a Fortran code used to transform EELS spectra into usable } information (according to D.B. Williams and C.B. Carter). - I do not have a } copy of the book, but could read it in a local University library or } purchase it.
*SNIP*
} Jerzy } } ***************************************************************** } Jerzy Gazda, Ph.D. } Advanced Micro Devices } Senior Materials Scientist } 5204 E. Ben White Blvd. - MS 613 } PCAL - Analytical TEM Section } Austin, TX 78741 } TEL: 1-800-538-8450, Ext. 51453 } FAX: (512) 602-7470 } jerzy,gazda-at-amd.com } *****************************************************************
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann 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-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
SCOPE The symposium is intended to provide an overview of present achievements and expected future trends in microscopy including the instrumentation, imaging science, theories, new techniques, new methods and applications in chemistry, physics, mineralogy, material sciences, life science, environmental science and other basic science areas. It will also cover scanning probe microscopy and near-field optical microscopy. Leading scientists will be invited to present introductory overview lectures to symposia. English will be the official language.
VENUE The symposium will take place in Kunming, the capital city of Yunnan Province and one of the most famous tourist destinations in China. Being blessed with the agreeable climate because of its subtropical location and altitude of 1894 meters, Kunming enjoys the fame of Spring City --- a city full of the beauty of spring all the year round. It is also fascinating due to the unique natural features such as the Stone Forest, Western Hill, Golden Temple and Ethnic Minorities Village. Therefore, Kunming has the great honor to be selected the host city of the 1999 International Horticultural Exposition (EXPO'99). The EXPO halls and gardens displaying flowers and plant life from all over the world certainly much highlight the city. They will last continuously after the EXPO'99.
Yunnan Province is located in South China bordering on Vietnam, Laos and Burma. The citizens are from 26 minority nationalities, each of which has her own language and folklore. Here people show a rich-color ethnic album by their unique history and culture, local customs and traditional festivals. The great variety of nationalities in Yunnan can be seen on the streets as well as in the EXPO Garden in Kunming. The southern atmosphere and the variety of cultures and customs can be enjoyed while strolling through the market streets away from the big avenues of the city.
INVITATION The Chinese Electron Microscopy Society (CEMS) would like to extend a warm invitation to all friends and colleagues in microscopy to join us at the '2000 International Kunming Symposium on Microscopy.
I would like to pick up on the expression `streaking'. Does this appear to be vertically from exposed areas? If there is insufficient agitation during developing this can occur quite easily. I assume that you develop the SO163 in concentrated developer and the 4489 in normal strength, maybe there is different agitation (N2 burst?) for the two films.
Other problems tend to give a blotchy or mottled effect.
} We are having problems with the developing of SO163 films. We get streaks } on the negatives and the results are highly inconsistent in terms of } exposure/optical density. We don't have these problems when we use 4489, } but we need the higher sensitivity of the SO163 for cryo work. Has anybody } else experienced such problems? } } I think at least some of these problems are related to the lack of temperature } control of our developing solutions. Now, we may be able to keep the developer } at the proper temperature by further cooling of the room with air-con. } However, is the temperature of the tap water used for rinsing also very } important? This is in Singapore, and our tap water is usually very warm, } sometimes close to 30C. } } If this is important, does anybody have experience with water cooling } systems for this purpose? A recirculating cooling water bath could be } used for cooling the developer, but wouldn't work for the rinse, for which } we would probably need some kind of in-line cooler. (Maybe similar to that } used in water drinking fountains?) I am concerned about the potentially } very high expense of such systems. } } Any advice would be greatly appreciated. Thanks, } } Terje } } } } ------------------------------------ } Dr. Terje Dokland } Senior Scientist } Institute of Molecular Agrobiology } 1 Research Link } The National University of Singapore } Singapore 117604 } Phone: 65-872 7405 Fax: 65-872 7007 } E-mail: dokland-at-ima.org.sg } http://lab.ima.org.sg/lab3052/terje } ------------------------------------ } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Ran into the same problems for a bit here too. Found it was warm tap water. The darkroom is maintained at 20C so we just stuck a bottle of water on the bench and problem solved. It was the rinse between development and fix that was the problem. We use 4489 film. Good luck
At 02:33 PM 9/27/1999 +0800, 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 "
Hello List Readers: Laura Rhoads suggested that forced use of THAT instrument would be a suitable penalty for those committing "crimes" such as broadcasting "dogma". I concured, adding "I would not want to offend anybody, but I've been in purgatory for two years already. Circa 1970 that instrument was the worst money could buy in the Western World."
Several readers have responded making positive points about that instruments. I agree with some of these because nothing in this world is entirely bad - even a broken clock shows the right time, twice a day.
Yes, that instrument looked good, it had a big column and would make a fine statue in someone's garden (so Laura suggested, personal communications). I would love one, it would be the crowing glory to the banks of the Ross River.
Yes, the image had good contrast because it was a medium resolution instrument (I guess about 0.8nm). A longer working distance objective results in higher contrast but lower resolution. Other manufacturers offer high contrast objectives, but most instrument purchasers ask for higher resolution.
Yes, RCA's are historical instrument and the first commercially produced American TEM. Though I believe that Siemens produced the first commercial TEM in the mid 30th.
Yes, for some people this was their first electron microscope and it had to be impressive to us who grew up before computer whizz-bangery. Unfortunately, it was my fourth TEM. My previous loves had been a mid-60th Siemens Elmiskop, a Philips 100C (the one with a near horizontal column and transmission viewing screen) and a Zeiss 9C. All of these were rather better suited to productive work.
Yes, the electronics were reliable, but the vacuum gauging was insufficient. Since it also had no vacuum locks and blanking provisions were poor, vacuum trouble-shooting was very difficult.
Yes, it was reasonably easy to operate and was great for forced coffee breaks. No specimen, no gun, nor camera vacuum locks made for lots and lots of pumping times.
Yes, the camera had two options, either three single plates could be inserted or one long plate taking five images on one plate. Bit of a pain to share negatives with another operator. Absolutely awful for taking lots of images and changing specimens. The pumping cycle was slow.
The complete lack of vacuum locks and the poor film options made the EMU-4 a "hysterical instrument". Such features were not rocket science even in those days. More features . . .
Changing the filament was a painful operation. I think the basic alignment after filament change required three complete column pumping cycles, lots of time for getting other jobs done during those operations. I could not align that scope in under two hours. I had inherited a service contract and the service men could do no better.
Alignment screws were difficult to adjust accurately in those days on most TEMs. I learned from the service man that the final touch was best achieved by tapping the column with a rubber mallet - which was not part of the service kit.
Filament life averaged about 12 hours, with the alignment procedure eating up a good part thereof. Short filament life may not have been an inherent problem in the design. This may have been due to poor vacuum, but there was no good way of isolating vacuum parts, getting a reliable vacuum reading and finding a possible leak.
Why did RCA sell the EMU-4 to Forgflow, who then quickly discontinued that instrument? Because the column design was 10 years too late. By 1972 I had a Philips EM300 - now that was progress. Philips at last had eclipsed the Elmiskops, which had been the top brand throughout the 50th and 60th.
Incidentally, Siemens closed their EM division about '78. In Karlsruhe in October '77 I had admired Siemens' latest "Wunderkind", it turned out be the Divisions death knell. It was a FESEM based somewhat on the research by Crewe (Chicago ?) Nice instrument, beautiful column, but Vacuum Generator of the UK had build a better performer at almost half the price. . . . the rest is history.
Disclaimer: Opinions expressed are mine they are based on fragile memories. No dogma is implied or intended. Feel free to purchase an EMU-4, if you can find one. Even Phantom comics of that period fetch increasing prices and modern instruments depreciate rapidly. Don't condemn me for another two years on THAT instrument please. PST does not supply the EMU-4 or similar. 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 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
I'd like to hear from anyone with experience with embedding and sectioning of cartilage for TEM. I've checked the "tips and tricks" page but don't see anything there specific to cartilage . Can anyone tell me if there are any particular pitfalls involved in this, especially in regards to maintaining good morphology the cells themselves? Fixation? Longer embedding times? Favorites resins? Sectioning? Thank you. Grace
Regarding AsCII from EL/P, go to the file menu and select "save as". A dialog box offers at least three types of ASCII.
I looked at the 1994 extended abstract on FANTOME. I noticed that the work was suported by DOE. That means that the program should be in the public domain. Since the primary author was at LBL and Roar Kilaas was mentioned in reference 5 (he is at LBL), try contacting LBL to see if they have the program.
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798 cook-at-aaem.amc.anl.gov
} Terje, We had a similar problem with SO163 that we solved by controlling agitation in the developer tank. We actually had streaks in our negatives because of excessive agitation. The nitrogen burst bubbler was not working and we used manual lifting of the rack to create the mixing that is required for uniform developing. Excessive lifting/agitation/mixing was causing "hot spots" due to the increased flow of the developer across the film in some regions, resulting in streaked images. We use a 4 minute developing time in diluted D-19. We now mix it manually, but slowly 3 or 4 times during the development, by smoothly lifting the rack of film out of the solution and tipping it in one direction and holding for a couple of seconds to briefly drain the developer back to the tank. The rack is then lowered smoothly back to the tank, and in 40-50 seconds the agitation is repeated but tipping the other way, holding briefly, and returning to the tank. This controlled agitation solved our problem. By the way, we also never had this problem when we used 4489, but opt for the more sensitive and finicky SO163. Brad Huggins BPAmoco Naperville, IL
} ---------- } From: Terje Dokland[SMTP:dokland-at-ima.org.sg] } Sent: Monday, September 27, 1999 1:33 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Developing of SO163 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } We are having problems with the developing of SO163 films. We get streaks } on the negatives and the results are highly inconsistent in terms of } exposure/optical density. We don't have these problems when we use 4489, } but we need the higher sensitivity of the SO163 for cryo work. Has anybody } else experienced such problems? } } I think at least some of these problems are related to the lack of } temperature } control of our developing solutions. Now, we may be able to keep the } developer } at the proper temperature by further cooling of the room with air-con. } However, is the temperature of the tap water used for rinsing also very } important? This is in Singapore, and our tap water is usually very warm, } sometimes close to 30C. } } If this is important, does anybody have experience with water cooling } systems for this purpose? A recirculating cooling water bath could be } used for cooling the developer, but wouldn't work for the rinse, for which } we would probably need some kind of in-line cooler. (Maybe similar to that } used in water drinking fountains?) I am concerned about the potentially } very high expense of such systems. } } Any advice would be greatly appreciated. Thanks, } } Terje } } } } ------------------------------------ } Dr. Terje Dokland } Senior Scientist } Institute of Molecular Agrobiology } 1 Research Link } The National University of Singapore } Singapore 117604 } Phone: 65-872 7405 Fax: 65-872 7007 } E-mail: dokland-at-ima.org.sg } http://lab.ima.org.sg/lab3052/terje } ------------------------------------ } } }
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Thank you, it's been a very interesting group. Marlene
To paraphrase an article from the Daily News (Burbank, Calif.) April 27, 1986. "They built the first commercial electron microscope to be produced in the United States." Richard Baker worked on the project from 1941 to 1947 with V.K. Zworykin and 2 other un-named scientists, at RCA, then located in Princeton, N.J. A benefactor of the Univ. of So. California purchased one of the instruments in 1946 and donated it to the university School of Medicine. A year later, Baker left RCA for USC and started doing research on the instrument he helped develop. This microscope, which was donated to the Smithsonian Institution, was replaced by the EMU-3? I had the honor of being trained in E.M. by Dr. Baker, and, yes, learning on the EMU. For more information, or a copy of the article, I can be reached at:
John Hardy City of Hope Medical Center E. M. Facility Duarte, CA. 91010 jhardy-at-coh.org (626) 301-8265
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
As Jim and Jan correctly pointed out, my original posting was meant to = stimulate a discussion about some of the issues facing biological electron = microscopists. =
Before I start, let me assure everyone that I had no intention of scaring = people from posting on this listserver. It has been, and still is, a = wonderful medium for everyone with similar interests. It is also an = important place to "publish" the information with no other place to go. I = have learned much from watching the public discussions and I regret that = much information exchange occurs off-line. My congratulations to all = readers, contributors and organizers for such a success. Please do not = let me put anyone off. If I have, then you must let me know so that I can = apologize and encourage you back.
Now on to the boring stuff:
When I decided to have some fun with what I consider to be a serious issue,= I chose my targets well. Not the innocent bystander, but experienced and = knowledgable experts in the field who are well qualified to put me down if = I stepped over the mark. A punishment I was willing to take if it was = proved I deserved it.
Molecular biologists "live and die by the protocol", which means that = their methods are so well defined that buying the book of recipes = techniques basically guarantees technical success.
For biological EM, we are less fortunate, and although I would enjoy the = simplicity of having set protocols, it is not yet possible. We are still = attempting to understand the many complex steps it takes to get "good" = micrographs. Understanding requires us to evaluate all the information we = have on each subject and, when needed, either design experiments to give = us more information or take what little we do understand and apply that = with little more than faith. =
As someone who is still learning, I crave for facts. When these are not = available, I can also live with statements that are qualified opinions and = work from these. However, seeing opinions being presented as fact worries = me. Not because I take them as truth, but because others do. =
I read the postings I picked on very carefully when they first came up. I = read them again recently and, although I paraphrased the content to make = my point, I still think they both qualified as statment of fact if read by = someone just starting in the field. If the postings had been softened = with "in my experience" or something similar to take the edge off, then I = think I would have let them go as is.
The off-line replies, supporting replies, and even the replies from the = original contributors, indicate a general support of my position. I = respect both Jim Darley and Jan Leunissen and have enjoyed their expertise = in contributions they have made to this list. I did not mean to make = hostile enemies of either one of them and I don't even disagree with the = facts of their psoting. My objection was in the presentation of the facts,= and their most recent messages sugegst that they also agree with this. = Here are my responses to the comments of Jan and Jim:
Jim, for high resolution work, there are better methods for immobilization = than aldehyde fixation. I wouldn't dream of examining any sample at high = magnification to look for resolution that just couldn't be there in resin-= embedded samples. Any comparison of fixation times is therefore not = important. As I said before, most of the extraction is a result of the = buffer being used and not the aldehyde. In fact, the best buffers for = preventing extraction (the "Good buffers": HEPES, PIPES etc) can retain = material so well that the tissue looks poorly fixed. The contrast people = aim at is more a result of extraction than addition of heavy metals. =
I must admit that I haven't come across your use of quotation marks to = mean a word isn't true. However, statments of generalization should be = qualified as such. =
For osmium fixation, the papers I quoted were chosen because, from what I = understand, there was no other way the amino acids under study could have = been localized. This is one example where a substantial exposure to = osmium tetroxide was required for localization to work. Other people sent = me more examples of this too. I routinely include osmium tetroxide in my = freeze substitution media on material that is destined for immunolabeling. = Under these low temperature conditions, the osmium does not appear to = affect either the labeling or the polymerization reaction.
This does not mean I would preferentially use osmium fixation and epoxy = resin embedding as a routine method for immunolabeling. However, to know = that for certain special instances, this method has worked better than = others, is another tool in my bag. As we all know, each new antibody we = work with and each new tissue or cell system requires us to think = carefully about our protocols. All are affected by what we know and the = more we know the better are our design parameters. I wouldn't keep new = microscopists from this information just because it is confusing. It is = because there is so much information available that we subscribe to this = listserver. =
All techniques have their uses and non should be excluded because of = generalities. In general, antibody labeling efficiency is better on = cryosections than on resin sections. However, for me to say that = cryosections should be used for immunolabeling experiments immediately = discounts the many advantages of resins and omits the important exceptions = where resins have been the only way to successfully localize antigens. We = have to know all our options so that we can make good decisions in our = experimental design. =
Jim, I almost did reply off-line to your posting in the way you suggested, = but I thought we could have a little more fun doing this way. Thinking = about these messages, and writing my replies has taken some time. I guess = you have had the same constraints of time so I apologize for dragging you = into it without warning.
Jan, it seems that you are also supporting my positions on dogma as well = as on the use of the listserver as an open forum, and on our right to make = informed decisions on which methods to use. =
As someone who is performing immunolabeling experiments on a daily basis, = I was fascinated by your comments on the glutaraldehyde fixation. The = lack of additional information made it difficult for me to properly = evaluate your statements and possible improve my protocols. I suppose I = could have contacted you directly to get this information but then it = would have been a little selfish and would have deprived us all of your = expertise.
Reading your humorous reply made me think again about my immunolabeling = protocols and convinced me to give the fixation step another try. However,= it seems we are both guilty of a similar crime in that our de=05cision to = either include or remove this fixation step in our labeling protocol was = based on subjective results and not on documented quantitative data. We = both saw what we wanted to see. So, until someone finds the time to = quantitatively compare results from both protocols, we sit with our = respective dogma's in our shared cell. =
What should we aim for? Perhaps protocols that are specific enough for = all systems. Or maybe protocols that explain a general example but which = also give the exceptions with explanations of why they worked against the = rule.
In the US, it is true that the accused remain innocent until proven guilty.= Without the time or resources to petition further, and therefore at risk = of adding boredom to the list by presenting incomplete research, I rest my = case with the accused retaining their innocence and liberty. =
I encourage suits for wrongful arrest from anyone interested in joining in = (I still think I escaped from this issue too easily). I also encourage = others to submit their comments on the general status of protocols for = biological EM. Do they sufficiently explain why we do them (e.g. how does = osmium tetroxide fix cells and why should this treatment modify antigens = enough to stop antibodies from binding to them?) or is there still some = black magic to these techniques? =
by srvr20.engin.umich.edu (8.9.1a/8.9.1) with ESMTP id PAA21653 for {microscopy-at-msa.microscopy.com} ; Mon, 27 Sep 1999 15:59:16 -0400 (EDT) X-Sender: bigelow-at-srvr5.engin.umich.edu Message-Id: {v03007802b4157bf9e6c3-at-[141.212.131.74]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I was working on a project a while ago that required the use of computer controlled stepping motors, and discovered a company that makes a complete package consisting of 2 stepping motors, power supply, and controller that is ready to connect to a PC for computer driven operation, and is VERY reasonably priced. Check with Circuit Specialists Inc. 800-811-5208, and ask about their MD2 system.
If you need a neat little minature stepping motor I found the Model AM1524 made by Donovan Micro-Tek, 805-522-4330 is only 15 mm in diameter and 16.5 mm long, and suitable for use inside a UHV system - maybe you could get Circuit Specialists to incorporate this motor into their system.
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
Very interesting history, which I hadn't known before. The RCA model at USC was probably replaced by the RCA EMU-2 series. I did EM for my PhD thesis in 1956-58 on an RCA EMU-2D, which was the current model then. I was working in Biology at Harvard with George B. Chapman, who had just arrived from Princeton, where he had worked with Jim Hillier (another member of that early RCA lab group).
I would appreciate a copy of the article. Thanks.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Anatomy and Cell Biology University of Michigan Medical School Ann Arbor, MI 48109-0616 Tel (work) 763-1287, Fax (work) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
--On Mon, Sep 27, 1999 11:40 AM -0500 "jhardy-at-smtplink.Coh.ORG"-at-sparc5.microscopy.com wrote:
} } To paraphrase an article from the Daily News (Burbank, Calif.) April } 27, 1986. "They built the first commercial electron microscope to } be produced in the United States." Richard Baker worked on the } project from 1941 to 1947 with V.K. Zworykin and 2 other un-named } scientists, at RCA, then located in Princeton, N.J. A benefactor } of the Univ. of So. California purchased one of the instruments in } 1946 and donated it to the university School of Medicine. A year } later, Baker left RCA for USC and started doing research on the } instrument he helped develop. This microscope, which was donated } to the } Smithsonian Institution, was replaced by the EMU-3? I had the honor } of being trained in E.M. by Dr. Baker, and, yes, learning on the EMU. } For more information, or a copy of the article, I can be reached at: } } John Hardy } City of Hope Medical Center } E. M. Facility } Duarte, CA. 91010 } jhardy-at-coh.org } (626) 301-8265
We have a Plasma Sciences LVC-76 Sputter coater with Carbon attachment that is having a mid-life crisis. It seems like some type of electrical or wiring problem, or perhaps a faulty switch?? but it works sporadically. (I am passing this on for a colleague, so am a bit fuzzy on the details). At least once the coating discharged towards the grounding wire inside the unit instead of evenly. Our internal electronics service personnel have exhausted their expertise towards repair.
Any suggestions on where to turn to have this unit repaired? I understand Plasma Sciences is no longer in existence. Alternatively, any suggestions on a good model to purchase these days? Preferably a company that will be around for a while (we've been through three different sputter coaters in the last 8 years, all requiring service, at least two of them from companies no longer in business.)
Thanks for your help! -- Karen S. Zaruba kszaruba-at-mmm.com 3M Center St. Paul, MN 55144
Hi Paul & Listreaders: I accept that you cited the three examples of dogma taunting to stir some debate. At the very least you picked rather poor examples of "dogma". The third of your "dogmas" was submitted by Jan Leunissen and he presented an excellent plea showing his innocence and also that he was not properly quoted. Since Jan works for Aurion I assume that he knows a few things about immuno gold labeling, yet Paul had stated } without producing supporting documentation or direct personal experiences. {
Paul's quotation accusing me of dogma were incorrect too. } "Can't leave biological tissues in glutaraldehyde for long periods; { I actually had clearly qualified this as referring to HIGH RESOLUTION work AND I had given reason AND I have experience in that regard. Paul, have you looked at GA overfixed tissue at several hundred thousand times and compared that with properly fixed tissues? I stand by my original statement.
Paul's second misquote was: } Immunological and cytochemical reactions are killed by osmium; { A little, but vital omission were the quotation marks around the word killed. Afterall, we are talking about Immunological and cystochemical activities and these are not on-or-off, but diminish with various treatments. My original advise was like saying: "70% ethanol kills microorganisms". It does and most would be killed very rapidly, but there may be some, for instance yeast spores that could survive for some time in that high concentration of yeast "excreta". The statement "70% ethanol 'kills' . . . remains true. I did not say or imply that osmium "kills" all immune activities instantly and completely. I am not guilty of "dogma", which is a forcible statement or opinion stating as if unchallengable or authoritative assumptions rather than empirical observations.
It should be noted that a wrongly based forcible statement like Paul's "Dogma Police" actually suits that very definitions much better. I also see a couple of other problems with this kind of argument on the listserver. 1 It could deter some people from submitting freely their opinions. 2 It could lead to the frequent addition and confusing qualifiers and this would change the nature of this discussion forum.
Hands up, which of the other contributors to the topic did go back to my original message and read this and Paul's assertions critically? Anybody??? The replies come up with a few publications showing that at least some partially osmicated tissues, retained some antigens in some cases. Not many such publications though and we could safely assume that less than one in a hundred of such research projects used any osmium prior to labeling. This supports my case: Why would so many researchers publish such low contrast images IF normal EM preparation showed equally good activities? Paul agrees } When we all finally accept that contrast equals loss of antigen, we may eventually enjoy the esthetics of low contrast, high information images, and learn to leave the high contrast pictures in the morphology books. { } From that I infer that at least some of those "osmium before labeling" publications may have been "better" without osmium first. Furthermore, Paul here clearly states that higher contrast (e.g. osmication) equals lower antigenicity; in fact he is in agreement with my original advise. What as your point?
I admit that I too played a game. Paul used an emotive term to get people onsite. It worked, because we all detest "dogma". It would have been nice if critical thinking had prevailed. When I asked to provide prove that osmication does not "kill" immune activity {cite some publications were full osmium fixation was used prior to immunolabeling {, they came in like the tide.
You cannot prove that by showing that some activity is preserved in some case. What is required is a full comparison of remaining activity after different treatments in different tissues and different reactions. Safe your efforts, Paul and most of as know that: osmium "kills" immune activities.
Now, had somebody in reply to my original submission written in "your right Jim, but sometimes, after some osmium fixation there is enough remaining activity for successful labeling." I would have replied "Amen". 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 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, September 24, 1999 4:42 AM, Paul Webster [SMTP:pwebster-at-mailhouse.hei.org] 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 } -----------------------------------------------------------------------. } } } Reply to: RE: Dogma Police } Dear Jim, } Thanks for your reply. I had hoped for more hostile and numerous responses } but my mailbox is only full of support messages. I didn't mean the message } to be a personal attack, rather a way of provoking an open discussion, } something that is often lacking on this forum. } My message was prompted mostly by the thread concerning publishing information } on-line, where the main objection was that there was insufficient review } process to make it credible. These comments followed by the solid statments } I quoted (admittedly out of context) clearly illustrated the point of what } poor refereeing can result in. } } I agree that for teaching purposes, a little dogma is not a bad thing and have } been guilty of it myself. However, to see you quote text in your web site as } a support for your claims made me think we should start discussing the finer } details of what we are doing here. } } Firstly, some references where osmium fixation has been used for } immunolabeling. Ottersen's group in Olso (1984 J. Comp Neurol 229:374-392; } 1986 Med. Biol. 64:147-158; 1990 Prog Brain Res 83:99-114) applied antibodies } to brain sections that had been fixed in 2.5% glutaraldehyde, 1% } formaldehyde, treated with 1% osmium tetroxide, dehydrated in ethanol and } propylene oxide and embedded in epoxy resin! I am sure I could find more } without too much effort if you want them (I remember what I read, not where I } read it). } } I see Tamara Howard has provided additional additional references and the } comments that there are few very hard and fast rules in EM specimen } preparation. I agree totally with this. It is important that we keep } attempting to try out new things so that we can push the bounderies. Setting } limits during the early stages of instruction stops growth into areas we can } never imagine. } } Jim, your other comment, that long fixation affects resolution is also not } documented fact. I do remember a paper, which I will look for, where } extraction in aldehyde fixative was dependant upon the buffer being used, not } the aldehyde. What is clearly documented is that the subsequent processing } steps (post-fixation, dehydration and infiltration) affect the amount of } extraction and thus reduce resolution. } For most morphology applications, contrast is preferred over resolution } anyway. Witness the problem of getting immunocytochemical results published } over the last 10 years. Even here on the listserver, there is always an } ongoing discussion of how to get better contrast in the LR or Lowicryl } resins. When we all finally accept that contrast equals loss of antigen, we } may eventually enjoy the esthetics of low contrast, high information images, } and learn to leave the high contrast pictures in the morphology books. } } If high resolution is required then there are better methods than those } involving aldehyde fixation and epoxy resin embedding. For the poster of the } original question I would say, leave the tissues in glutaraldehyde for as } long as is convenient, any loss of structure will go unnoticed. However, as } with all advice, I reccommend you take it with a pinch of salt and try it out } for yourself. } } Finally, the issue of fixing immunolabeled sections with 1% glutaraldehyde } came up and there were a variety of answers. The one I took mild exception } to was from Jan Leunissen. He is a great teacher and I respect him and his } work at many levels. In his message, he correctly pointed out that the } conditions of high salt and low pH are used to elute antibodies from } antigens. Therefore 1% glutaraldehyde must be used to crosslink antibodies } to sections and protein A-gold to antibodies. It all sounds good in theory } but where is the proof? I and my colleagues have been applying antibodies } and colloidal gold to sections for years without that final crosslinking } step. Why do we still get specific label? Who knows. What we do know is } that we have compared the effect of adding this fixation step and decided } that it didn't improve labeling efficiency. We therefore made an informed } decision to leave it out of the protocol. Putting unneccessary steps into a } protocol only makes life more difficult for people wo } rking in busy labs. } My advice for anyone just starting in EM, is to take what you are told with } healthy skepticism and question anything you feel may be unreasonable. If } there is a reason for doing something your teacher should know that reason. } If they don't know why something should be done, try leaving it out to see } if it makes a difference. To apply methods on the faith that they work is } not sufficient anymore. We need informed reasons for applying methods. We } are in the good times for EM and we have to shed our artist/magician image. } The closer we get to being scientists, the more seriously we will be } accepted by the science community who are at present looking for our skills. } } For the record, I also take exception to the methods that involve the use of } bodily fluids. If we can't work out how to do something without nose or face } grease we really deserve to be left behind. } } Thanks Jim, for taking the bait and allowing the start of what I hope will be } an informative discussion on credibility. } } Regards, } } Paul Webster. } } } } } jim wrote: } } Fair go Paul. That "Dogma Police" header is a trifle emotive and not } } justified? } The first two examples of "Dogma" given may be mine, but they are } } out of } context. } } I had written that storing tissues in GA affects high resolution imaging. I } } } gave reasons. } } Are you asserting that excessive fixation in GA does not affect high } } resolution } imaging??? } } } } I had written {Immunological and cytochemical reactions are killed by } } osmium} , } but the "killed" was in quotation marks, because I had learned } } about 30 years } ago, that most such cellular activities cease . I do not } } believe that cells or } osmium have changed in this regard. I have no reason } } to change my mind - or do } you have any evidence to the contrary. I guess } } about half of all subscribers } would like to know about that. To use your } } words "were is your supporting } documentation" Please Paul, cite some } } publications were full osmium fixation } was used prior to immunolabeling. } } 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 } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } } } } On Thursday, September 23, 1999 2:17 AM, Paul Webster } } } [SMTP:pwebster-at-mailhouse.hei.org] wrote: } } } } } "Can't leave biological tissues in glutaraldehyde for long periods; } } } Immunological and cytochemical reactions are killed by osmium; } } } Must use glutaradehyde after labeling reactions on sections." } } } } } } You are all under arrest for spreading widespread panic without producing } } } supporting documentation or direct personal experiences. } } } } } } Paul Webster, Ph.D. } } } House Ear Insititute } } } 2100 West Third Street } } } Los Angeles, CA 90057 } } } 213 273 8026 } } } pwebster-at-hei.org } } } http://www.hei.org/htm/aemi.htm } } } } } } } } } } } } } RFC822 header } } ----------------------------------- } } } } Received: from ultra.ultra.net.au [203.20.237.5] by mailhouse.hei.org with } } } ESMTP } } (SMTPD32-4.07) id A00314BB021E; Thu, 23 Sep 1999 07:58:11 PST } } Received: from 150 (p098.supa2-tsv.ultra.net.au [202.80.71.98]) } } by ultra.ultra.net.au (8.9.3/8.9.3) with SMTP id BAA13065; } } Fri, 24 Sep 1999 01:02:04 +1000 (EST) } } Received: by localhost with Microsoft MAPI; Fri, 24 Sep 1999 01:02:01 +1000 } } Message-ID: {01BF0628.68BC69A0.jim-at-proscitech.com.au} } } From: jim {jim-at-proscitech.com.au} } } Reply-To: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } } To: "'Paul Webster'" {pwebster-at-mailhouse.hei.org} , } } "'MSA listserver submission'" } } {Microscopy-at-sparc5.microscopy.com} } } Subject: RE: Dogma Police } } Date: Fri, 24 Sep 1999 00:50:03 +1000 } } Return-Receipt-To: jim {jim-at-proscitech.com.au} } } Organization: ProSciTech } } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } MIME-Version: 1.0 } } Content-Type: text/plain; charset="us-ascii" } } Content-Transfer-Encoding: 7bit } } X-UIDL: 234843928 } } Status: U } } }
We are having problems with the developing of SO163 films. We get streaks on the negatives and the results are highly inconsistent in terms of exposure/optical density. We don't have these problems when we use 4489, but we need the higher sensitivity of the SO163 for cryo work. Has anybody else experienced such problems?
I think at least some of these problems are related to the lack of temperature control of our developing solutions. Now, we may be able to keep the developer at the proper temperature by further cooling of the room with air-con. However, is the temperature of the tap water used for rinsing also very important? This is in Singapore, and our tap water is usually very warm, sometimes close to 30C.
If this is important, does anybody have experience with water cooling systems for this purpose? A recirculating cooling water bath could be used for cooling the developer, but wouldn't work for the rinse, for which we would probably need some kind of in-line cooler. (Maybe similar to that used in water drinking fountains?) I am concerned about the potentially very high expense of such systems.
Any advice would be greatly appreciated. Thanks,
Terje
------------------------------------ Dr. Terje Dokland Senior Scientist Institute of Molecular Agrobiology 1 Research Link The National University of Singapore Singapore 117604 Phone: 65-872 7405 Fax: 65-872 7007 E-mail: dokland-at-ima.org.sg http://lab.ima.org.sg/lab3052/terje ------------------------------------
Hi everybody,
I'm a bit late in this discussion, have been trying to get on the list for a while -
I've been using Lowicryl K4M for nearly 10 years now, and only a few times have I had problems with sectioning. I can't recall ever having problems with the curing either (which have been the case with both Agar 100 and Araldite). I tried Unicryl for a while as well (both heat- and UV-polymerized), and my results were quite comparable with the Lowicryl ones. Has anyone compared Low/Uni with LR White or Gold?
Regards Margareta
Margareta Halin Dept. of Internal Medicine University Hospital Uppsala Sweden
Ran into the same problems for a bit here too. Found it was warm tap water. The darkroom is maintained at 20C so we just stuck a bottle of water on the bench and problem solved. It was the rinse between development and fix that was the problem. We use 4489 film. Good luck
At 02:33 PM 9/27/1999 +0800, 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 "
Dear Jerzy,
Mr. Egerton placed all the source codes that are discussed in his book and some other helpful files and data on the following ftp-site
ftp://ftp.phys.ualberta.ca/public_html/eels/
Cheers,
Petra
At 19:54 24.09.99 -0500, you wrote:
*SNIP*
} I am attempting to conduct dielectric function (constant) evaluations from } Plasmon Loss EELS spectra acquired with our TEM using GIF. After doing a } fast literature search I came up with two possible sources of information: } } 1) book by Egerton (1996 2nd edition) is supposed to contain recipe for } calculations and a Fortran code used to transform EELS spectra into usable } information (according to D.B. Williams and C.B. Carter). - I do not have a } copy of the book, but could read it in a local University library or } purchase it.
*SNIP*
} Jerzy } } ***************************************************************** } Jerzy Gazda, Ph.D. } Advanced Micro Devices } Senior Materials Scientist } 5204 E. Ben White Blvd. - MS 613 } PCAL - Analytical TEM Section } Austin, TX 78741 } TEL: 1-800-538-8450, Ext. 51453 } FAX: (512) 602-7470 } jerzy,gazda-at-amd.com } *****************************************************************
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann 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-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
SCOPE The symposium is intended to provide an overview of present achievements and expected future trends in microscopy including the instrumentation, imaging science, theories, new techniques, new methods and applications in chemistry, physics, mineralogy, material sciences, life science, environmental science and other basic science areas. It will also cover scanning probe microscopy and near-field optical microscopy. Leading scientists will be invited to present introductory overview lectures to symposia. English will be the official language.
VENUE The symposium will take place in Kunming, the capital city of Yunnan Province and one of the most famous tourist destinations in China. Being blessed with the agreeable climate because of its subtropical location and altitude of 1894 meters, Kunming enjoys the fame of Spring City --- a city full of the beauty of spring all the year round. It is also fascinating due to the unique natural features such as the Stone Forest, Western Hill, Golden Temple and Ethnic Minorities Village. Therefore, Kunming has the great honor to be selected the host city of the 1999 International Horticultural Exposition (EXPO'99). The EXPO halls and gardens displaying flowers and plant life from all over the world certainly much highlight the city. They will last continuously after the EXPO'99.
Yunnan Province is located in South China bordering on Vietnam, Laos and Burma. The citizens are from 26 minority nationalities, each of which has her own language and folklore. Here people show a rich-color ethnic album by their unique history and culture, local customs and traditional festivals. The great variety of nationalities in Yunnan can be seen on the streets as well as in the EXPO Garden in Kunming. The southern atmosphere and the variety of cultures and customs can be enjoyed while strolling through the market streets away from the big avenues of the city.
INVITATION The Chinese Electron Microscopy Society (CEMS) would like to extend a warm invitation to all friends and colleagues in microscopy to join us at the '2000 International Kunming Symposium on Microscopy.
NOTICE: We shall keep you apprised of the further information when you name be listed.
(This is the END of the announcement.)
Dear Terge,
I would like to pick up on the expression `streaking'. Does this appear to be vertically from exposed areas? If there is insufficient agitation during developing this can occur quite easily. I assume that you develop the SO163 in concentrated developer and the 4489 in normal strength, maybe there is different agitation (N2 burst?) for the two films.
Other problems tend to give a blotchy or mottled effect.
} We are having problems with the developing of SO163 films. We get streaks } on the negatives and the results are highly inconsistent in terms of } exposure/optical density. We don't have these problems when we use 4489, } but we need the higher sensitivity of the SO163 for cryo work. Has anybody } else experienced such problems? } } I think at least some of these problems are related to the lack of temperature } control of our developing solutions. Now, we may be able to keep the developer } at the proper temperature by further cooling of the room with air-con. } However, is the temperature of the tap water used for rinsing also very } important? This is in Singapore, and our tap water is usually very warm, } sometimes close to 30C. } } If this is important, does anybody have experience with water cooling } systems for this purpose? A recirculating cooling water bath could be } used for cooling the developer, but wouldn't work for the rinse, for which } we would probably need some kind of in-line cooler. (Maybe similar to that } used in water drinking fountains?) I am concerned about the potentially } very high expense of such systems. } } Any advice would be greatly appreciated. Thanks, } } Terje } } } } ------------------------------------ } Dr. Terje Dokland } Senior Scientist } Institute of Molecular Agrobiology } 1 Research Link } The National University of Singapore } Singapore 117604 } Phone: 65-872 7405 Fax: 65-872 7007 } E-mail: dokland-at-ima.org.sg } http://lab.ima.org.sg/lab3052/terje } ------------------------------------ } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Hello List Readers: Laura Rhoads suggested that forced use of THAT instrument would be a suitable penalty for those committing "crimes" such as broadcasting "dogma". I concured, adding "I would not want to offend anybody, but I've been in purgatory for two years already. Circa 1970 that instrument was the worst money could buy in the Western World."
Several readers have responded making positive points about that instruments. I agree with some of these because nothing in this world is entirely bad - even a broken clock shows the right time, twice a day.
Yes, that instrument looked good, it had a big column and would make a fine statue in someone's garden (so Laura suggested, personal communications). I would love one, it would be the crowing glory to the banks of the Ross River.
Yes, the image had good contrast because it was a medium resolution instrument (I guess about 0.8nm). A longer working distance objective results in higher contrast but lower resolution. Other manufacturers offer high contrast objectives, but most instrument purchasers ask for higher resolution.
Yes, RCA's are historical instrument and the first commercially produced American TEM. Though I believe that Siemens produced the first commercial TEM in the mid 30th.
Yes, for some people this was their first electron microscope and it had to be impressive to us who grew up before computer whizz-bangery. Unfortunately, it was my fourth TEM. My previous loves had been a mid-60th Siemens Elmiskop, a Philips 100C (the one with a near horizontal column and transmission viewing screen) and a Zeiss 9C. All of these were rather better suited to productive work.
Yes, the electronics were reliable, but the vacuum gauging was insufficient. Since it also had no vacuum locks and blanking provisions were poor, vacuum trouble-shooting was very difficult.
Yes, it was reasonably easy to operate and was great for forced coffee breaks. No specimen, no gun, nor camera vacuum locks made for lots and lots of pumping times.
Yes, the camera had two options, either three single plates could be inserted or one long plate taking five images on one plate. Bit of a pain to share negatives with another operator. Absolutely awful for taking lots of images and changing specimens. The pumping cycle was slow.
The complete lack of vacuum locks and the poor film options made the EMU-4 a "hysterical instrument". Such features were not rocket science even in those days. More features . . .
Changing the filament was a painful operation. I think the basic alignment after filament change required three complete column pumping cycles, lots of time for getting other jobs done during those operations. I could not align that scope in under two hours. I had inherited a service contract and the service men could do no better.
Alignment screws were difficult to adjust accurately in those days on most TEMs. I learned from the service man that the final touch was best achieved by tapping the column with a rubber mallet - which was not part of the service kit.
Filament life averaged about 12 hours, with the alignment procedure eating up a good part thereof. Short filament life may not have been an inherent problem in the design. This may have been due to poor vacuum, but there was no good way of isolating vacuum parts, getting a reliable vacuum reading and finding a possible leak.
Why did RCA sell the EMU-4 to Forgflow, who then quickly discontinued that instrument? Because the column design was 10 years too late. By 1972 I had a Philips EM300 - now that was progress. Philips at last had eclipsed the Elmiskops, which had been the top brand throughout the 50th and 60th.
Incidentally, Siemens closed their EM division about '78. In Karlsruhe in October '77 I had admired Siemens' latest "Wunderkind", it turned out be the Divisions death knell. It was a FESEM based somewhat on the research by Crewe (Chicago ?) Nice instrument, beautiful column, but Vacuum Generator of the UK had build a better performer at almost half the price. . . . the rest is history.
Disclaimer: Opinions expressed are mine they are based on fragile memories. No dogma is implied or intended. Feel free to purchase an EMU-4, if you can find one. Even Phantom comics of that period fetch increasing prices and modern instruments depreciate rapidly. Don't condemn me for another two years on THAT instrument please. PST does not supply the EMU-4 or similar. 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 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
} I have just been asked to come up with a system for automatically } replenishing the liquid nitrogen in the anticontamination device
Hi Scott,
One of our customers recommended Vacuum Barrier Systems (VBS). Website is {http://www.vbsflex.com} .
Jeff Gschwend EDAX, Inc.
Phone: 847-816-6098
I'd like to hear from anyone with experience with embedding and sectioning of cartilage for TEM. I've checked the "tips and tricks" page but don't see anything there specific to cartilage . Can anyone tell me if there are any particular pitfalls involved in this, especially in regards to maintaining good morphology the cells themselves? Fixation? Longer embedding times? Favorites resins? Sectioning? Thank you. Grace
To paraphrase an article from the Daily News (Burbank, Calif.) April 27, 1986. "They built the first commercial electron microscope to be produced in the United States." Richard Baker worked on the project from 1941 to 1947 with V.K. Zworykin and 2 other un-named scientists, at RCA, then located in Princeton, N.J. A benefactor of the Univ. of So. California purchased one of the instruments in 1946 and donated it to the university School of Medicine. A year later, Baker left RCA for USC and started doing research on the instrument he helped develop. This microscope, which was donated to the Smithsonian Institution, was replaced by the EMU-3? I had the honor of being trained in E.M. by Dr. Baker, and, yes, learning on the EMU. For more information, or a copy of the article, I can be reached at:
John Hardy City of Hope Medical Center E. M. Facility Duarte, CA. 91010 jhardy-at-coh.org (626) 301-8265
} Terje, We had a similar problem with SO163 that we solved by controlling agitation in the developer tank. We actually had streaks in our negatives because of excessive agitation. The nitrogen burst bubbler was not working and we used manual lifting of the rack to create the mixing that is required for uniform developing. Excessive lifting/agitation/mixing was causing "hot spots" due to the increased flow of the developer across the film in some regions, resulting in streaked images. We use a 4 minute developing time in diluted D-19. We now mix it manually, but slowly 3 or 4 times during the development, by smoothly lifting the rack of film out of the solution and tipping it in one direction and holding for a couple of seconds to briefly drain the developer back to the tank. The rack is then lowered smoothly back to the tank, and in 40-50 seconds the agitation is repeated but tipping the other way, holding briefly, and returning to the tank. This controlled agitation solved our problem. By the way, we also never had this problem when we used 4489, but opt for the more sensitive and finicky SO163. Brad Huggins BPAmoco Naperville, IL
} ---------- } From: Terje Dokland[SMTP:dokland-at-ima.org.sg] } Sent: Monday, September 27, 1999 1:33 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Developing of SO163 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } We are having problems with the developing of SO163 films. We get streaks } on the negatives and the results are highly inconsistent in terms of } exposure/optical density. We don't have these problems when we use 4489, } but we need the higher sensitivity of the SO163 for cryo work. Has anybody } else experienced such problems? } } I think at least some of these problems are related to the lack of } temperature } control of our developing solutions. Now, we may be able to keep the } developer } at the proper temperature by further cooling of the room with air-con. } However, is the temperature of the tap water used for rinsing also very } important? This is in Singapore, and our tap water is usually very warm, } sometimes close to 30C. } } If this is important, does anybody have experience with water cooling } systems for this purpose? A recirculating cooling water bath could be } used for cooling the developer, but wouldn't work for the rinse, for which } we would probably need some kind of in-line cooler. (Maybe similar to that } used in water drinking fountains?) I am concerned about the potentially } very high expense of such systems. } } Any advice would be greatly appreciated. Thanks, } } Terje } } } } ------------------------------------ } Dr. Terje Dokland } Senior Scientist } Institute of Molecular Agrobiology } 1 Research Link } The National University of Singapore } Singapore 117604 } Phone: 65-872 7405 Fax: 65-872 7007 } E-mail: dokland-at-ima.org.sg } http://lab.ima.org.sg/lab3052/terje } ------------------------------------ } } }
Regarding AsCII from EL/P, go to the file menu and select "save as". A dialog box offers at least three types of ASCII.
I looked at the 1994 extended abstract on FANTOME. I noticed that the work was suported by DOE. That means that the program should be in the public domain. Since the primary author was at LBL and Roar Kilaas was mentioned in reference 5 (he is at LBL), try contacting LBL to see if they have the program.
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798 cook-at-aaem.amc.anl.gov
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When using manual agitation every minute or so with Diafine two-bath developer, I've never see any streaking on the dried negatives.
Bernie Kestel Materials Science Division Argonne National Laboratory 9700 South Cass Avenue Argonne,Il., 60439
unsubcribe mheller-at-u.washington.edu
Thank you, it's been a very interesting group. Marlene
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As Jim and Jan correctly pointed out, my original posting was meant to stimulate a discussion about some of the issues facing biological electron microscopists. Before I start, let me assure everyone that I had no intention of scaring people from posting on this listserver. It has been, and still is, a wonderful medium for everyone with similar interests. It is also an important place to "publish" the information with no other place to go. I have learned much from watching the public discussions and I regret that much information exchange occurs off-line. My congratulations to all readers, contributors and organizers for such a success. Please do not let me put anyone off. If I have, then you must let me know so that I can apologize and encourage you back.
Now on to the boring stuff:
When I decided to have some fun with what I consider to be a serious issue, I chose my targets well. Not the innocent bystander, but experienced and knowledgable experts in the field who are well qualified to put me down if I stepped over the mark. A punishment I was willing to take if it was proved I deserved it.
Molecular biologists "live and die by the protocol", which means that their methods are so well defined that buying the book of recipes techniques basically guarantees technical success.
For biological EM, we are less fortunate, and although I would enjoy the simplicity of having set protocols, it is not yet possible. We are still attempting to understand the many complex steps it takes to get "good" micrographs. Understanding requires us to evaluate all the information we have on each subject and, when needed, either design experiments to give us more information or take what little we do understand and apply that with little more than faith. As someone who is still learning, I crave for facts. When these are not available, I can also live with statements that are qualified opinions and work from these. However, seeing opinions being presented as fact worries me. Not because I take them as truth, but because others do. I read the postings I picked on very carefully when they first came up. I read them again recently and, although I paraphrased the content to make my point, I still think they both qualified as statment of fact if read by someone just starting in the field. If the postings had been softened with "in my experience" or something similar to take the edge off, then I think I would have let them go as is.
The off-line replies, supporting replies, and even the replies from the original contributors, indicate a general support of my position. I respect both Jim Darley and Jan Leunissen and have enjoyed their expertise in contributions they have made to this list. I did not mean to make hostile enemies of either one of them and I don't even disagree with the facts of their psoting. My objection was in the presentation of the facts, and their most recent messages sugegst that they also agree with this. Here are my responses to the comments of Jan and Jim:
Jim, for high resolution work, there are better methods for immobilization than aldehyde fixation. I wouldn't dream of examining any sample at high magnification to look for resolution that just couldn't be there in resin-embedded samples. Any comparison of fixation times is therefore not important. As I said before, most of the extraction is a result of the buffer being used and not the aldehyde. In fact, the best buffers for preventing extraction (the "Good buffers": HEPES, PIPES etc) can retain material so well that the tissue looks poorly fixed. The contrast people aim at is more a result of extraction than addition of heavy metals. I must admit that I haven't come across your use of quotation marks to mean a word isn't true. However, statments of generalization should be qualified as such. For osmium fixation, the papers I quoted were chosen because, from what I understand, there was no other way the amino acids under study could have been localized. This is one example where a substantial exposure to osmium tetroxide was required for localization to work. Other people sent me more examples of this too. I routinely include osmium tetroxide in my freeze substitution media on material that is destined for immunolabeling. Under these low temperature conditions, the osmium does not appear to affect either the labeling or the polymerization reaction.
This does not mean I would preferentially use osmium fixation and epoxy resin embedding as a routine method for immunolabeling. However, to know that for certain special instances, this method has worked better than others, is another tool in my bag. As we all know, each new antibody we work with and each new tissue or cell system requires us to think carefully about our protocols. All are affected by what we know and the more we know the better are our design parameters. I wouldn't keep new microscopists from this information just because it is confusing. It is because there is so much information available that we subscribe to this listserver. All techniques have their uses and non should be excluded because of generalities. In general, antibody labeling efficiency is better on cryosections than on resin sections. However, for me to say that cryosections should be used for immunolabeling experiments immediately discounts the many advantages of resins and omits the important exceptions where resins have been the only way to successfully localize antigens. We have to know all our options so that we can make good decisions in our experimental design. Jim, I almost did reply off-line to your posting in the way you suggested, but I thought we could have a little more fun doing this way. Thinking about these messages, and writing my replies has taken some time. I guess you have had the same constraints of time so I apologize for dragging you into it without warning.
Jan, it seems that you are also supporting my positions on dogma as well as on the use of the listserver as an open forum, and on our right to make informed decisions on which methods to use. As someone who is performing immunolabeling experiments on a daily basis, I was fascinated by your comments on the glutaraldehyde fixation. The lack of additional information made it difficult for me to properly evaluate your statements and possible improve my protocols. I suppose I could have contacted you directly to get this information but then it would have been a little selfish and would have deprived us all of your expertise.
Reading your humorous reply made me think again about my immunolabeling protocols and convinced me to give the fixation step another try. However, it seems we are both guilty of a similar crime in that our decision to either include or remove this fixation step in our labeling protocol was based on subjective results and not on documented quantitative data. We both saw what we wanted to see. So, until someone finds the time to quantitatively compare results from both protocols, we sit with our respective dogma's in our shared cell. What should we aim for? Perhaps protocols that are specific enough for all systems. Or maybe protocols that explain a general example but which also give the exceptions with explanations of why they worked against the rule.
In the US, it is true that the accused remain innocent until proven guilty. Without the time or resources to petition further, and therefore at risk of adding boredom to the list by presenting incomplete research, I rest my case with the accused retaining their innocence and liberty. I encourage suits for wrongful arrest from anyone interested in joining in (I still think I escaped from this issue too easily). I also encourage others to submit their comments on the general status of protocols for biological EM. Do they sufficiently explain why we do them (e.g. how does osmium tetroxide fix cells and why should this treatment modify antigens enough to stop antibodies from binding to them?) or is there still some black magic to these techniques? Regards,
Paul Webster.
I was working on a project a while ago that required the use of computer controlled stepping motors, and discovered a company that makes a complete package consisting of 2 stepping motors, power supply, and controller that is ready to connect to a PC for computer driven operation, and is VERY reasonably priced. Check with Circuit Specialists Inc. 800-811-5208, and ask about their MD2 system.
If you need a neat little minature stepping motor I found the Model AM1524 made by Donovan Micro-Tek, 805-522-4330 is only 15 mm in diameter and 16.5 mm long, and suitable for use inside a UHV system - maybe you could get Circuit Specialists to incorporate this motor into their system.
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
Very interesting history, which I hadn't known before. The RCA model at USC was probably replaced by the RCA EMU-2 series. I did EM for my PhD thesis in 1956-58 on an RCA EMU-2D, which was the current model then. I was working in Biology at Harvard with George B. Chapman, who had just arrived from Princeton, where he had worked with Jim Hillier (another member of that early RCA lab group).
I would appreciate a copy of the article. Thanks.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Anatomy and Cell Biology University of Michigan Medical School Ann Arbor, MI 48109-0616 Tel (work) 763-1287, Fax (work) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
--On Mon, Sep 27, 1999 11:40 AM -0500 "jhardy-at-smtplink.Coh.ORG"-at-sparc5.microscopy.com wrote:
} } To paraphrase an article from the Daily News (Burbank, Calif.) April } 27, 1986. "They built the first commercial electron microscope to } be produced in the United States." Richard Baker worked on the } project from 1941 to 1947 with V.K. Zworykin and 2 other un-named } scientists, at RCA, then located in Princeton, N.J. A benefactor } of the Univ. of So. California purchased one of the instruments in } 1946 and donated it to the university School of Medicine. A year } later, Baker left RCA for USC and started doing research on the } instrument he helped develop. This microscope, which was donated } to the } Smithsonian Institution, was replaced by the EMU-3? I had the honor } of being trained in E.M. by Dr. Baker, and, yes, learning on the EMU. } For more information, or a copy of the article, I can be reached at: } } John Hardy } City of Hope Medical Center } E. M. Facility } Duarte, CA. 91010 } jhardy-at-coh.org } (626) 301-8265
We have a Plasma Sciences LVC-76 Sputter coater with Carbon attachment that is having a mid-life crisis. It seems like some type of electrical or wiring problem, or perhaps a faulty switch?? but it works sporadically. (I am passing this on for a colleague, so am a bit fuzzy on the details). At least once the coating discharged towards the grounding wire inside the unit instead of evenly. Our internal electronics service personnel have exhausted their expertise towards repair.
Any suggestions on where to turn to have this unit repaired? I understand Plasma Sciences is no longer in existence. Alternatively, any suggestions on a good model to purchase these days? Preferably a company that will be around for a while (we've been through three different sputter coaters in the last 8 years, all requiring service, at least two of them from companies no longer in business.)
Thanks for your help! -- Karen S. Zaruba kszaruba-at-mmm.com 3M Center St. Paul, MN 55144
} Dear List, } } I am wondering if any of you in a university setting could offer some } prices ranges that you charge for the use of your electron microscopes, } and other services. We have outside clientele (non university) as well } as faculty/student usage that, either, have training and require no help } from the lab staff, or they have no experience and require staff to go
} through the entire process, including helping view. Any input, } including resources that you may have used to determine pricing, would
} be appreciated. } Thanks, } } Melissa Carter } Microscope and Graphic Imaging Center } California State Hayward University } Hayward, CA 94546
Attn all RCA-o-philes,
With regard to Jim Darley's citing of the High Crimes in the Diet of Dogma Court Record... } } Hello List Readers: } Laura Rhoads suggested that forced use of THAT instrument would be a suitable } penalty for those committing "crimes" such as broadcasting "dogma". I } concured, } adding "I would not want to offend anybody, but I've been in purgatory for } two } years already. Circa 1970 that instrument was the worst money could buy in } the } Western World." } } Yes, that instrument looked good, it had a big column and would make a fine } statue in someone's garden (so Laura suggested, personal communications). I } would love one, it would be the crowing glory to the banks of the Ross River.
Jim- You've got it all wrong: I said lawn ornament, not garden. If I put an EMU-4 column and requisite support pedestal in my garden there would be no room for my radishes... Actually, someone has already offered me an RCA EMT for this purpose, but that model isn't nearly massive enough for what I intend (I think it's complete so if anyone wants this artifact say, for a home workshop restoration project, contact me off-list).
Actually, the subject of RCA-for-pillory has brought up some rather interesting interpretations regarding the application of RCA EO equipment. Since I had yet to enter kindergarten by the time the last EMU-4 rolled off the assembly line in 1969 I never had the opportunity many others seem to have had cutting their teeth (I was cutting my own) on this (at the time) cutting edge technology. Missing this equipment in its prime, I feel somehow cheated, and as a result will have to rely on the experiences of those who did. What I find really interesting is the disparate variety of opinions as to service and operation. Could the EMU-4 Jim used at the World Expo have been a made on a Monday and Chuck Garber's, for example, a Wednesday? Or, did the RCA engineers install some extra parts they found in Hangar 18 from the Roswell, NM Crash Site that Jim's didn't get? Since the SPI rig is still running perhaps the operators can comment on their filament life and if it exceeds 12 hours? In order to solve the poor vacuum pumping problem (and to avoid forced coffee breaks) has this unit's vacuum system been retrofitted with a Balzers turbo pump perhaps? Are the alignment problems vividly recalled simply the result of using the wrong size mallet maybe?
All these issues are quite intriguing. If anyone does look into the history of RCA then maybe other manufacturers of EO equipment, such as Siemens, would be in order as well? And who was Vacuum Generator of the UK? Those who forget the past are doomed to repeat its failures. It seems that the technical information is available, and MSA has a tremendous body of institutional memory. It would be a shame to have this collective wisdom disappear.
For that matter, when I arrived at my latest job I discovered a complete Philips 75C hidden in the corner of the basement, with original warranty card (never filled out) which I am told still operates...Where will I ever get parts?...
************************************************ If the iron dice must roll, may God help us all...
Theobald von Bethmann-Hollweg German chancellor, August 1, 1914
************************************************ Laura Rhoads Biology Department SUNY Potsdam Potsdam, NY 13676 315-267-2260 315-267-3170 fax
I have been asked to prepare TS's of membrane filters for SEM. The filters are polysulphonate based with a polyethylene coating. They look like long tubes of paper Is there a simple way of cutting them to give a good clean surface? Any ideas appreciated.
Alfred Harris
Dear colleagues, I would like to bring this meeting to your attention:
THE 7TH ASIA-PACIFIC ELECTRON MICROSCOPY CONFERENCE "Perspective Imaging" Singapore International Convention and Exhibition Centre Singapore, 26-30 June 2000
The meeting covers all aspects of electron microscopy, including biological microscopy, material sciences, spectroscopic techniques, novel microscopies and image analysis, and there is a special symposium dedicated to 3D imaging of macromolecules, featuring a number of distinguished speakers like Wah Chiu, Ken Downing, Wolfgang Baumeister, Yoshi Fujiyoshi and many others.
As a venue, Singapore provides an exciting international atmosphere combined with a strong local flavour, including delicious cuisine, tropical climate and ample opportunities to explore the surrounding region, such as Thailand, Bali and Malaysia.
Further information about the meeting and about Singapore can be found at the APEM2000 web site at http://www.med.nus.edu.sg/micsoc/7apem/, or contact the conference secretariat at antbaybh-at-nus.edu.sg or micngml-at-nus.edu.sg.
Please note that deadline for abstracts is 31 December 1999; deadline for advance registration is 31 January 2000, and final date for securing accommodation is 12 June 2000.
See you in Singapore in 2000!
------------------------------------ Dr. Terje Dokland Senior Scientist Institute of Molecular Agrobiology 1 Research Link The National University of Singapore Singapore 117604 Phone: 65-872 7405 Fax: 65-872 7007 E-mail: dokland-at-ima.org.sg http://lab.ima.org.sg/lab3052/terje ------------------------------------
Alfred,
In answer to your question:
} I have been asked to prepare TS's of membrane filters for SEM. The filters } are polysulphonate based with a polyethylene coating. They look like long } tubes of paper Is there a simple way of cutting them to give a good clean } surface? Any ideas appreciated.
If you were to cut them under liquid nitrogen, you would get a much cleaner fracture surface. Another thing might be to infiltrate the filter with a low viscosity curing resin, outgas in a vacuum oven, and cure. (Someone else on the listserver could perhaps recommend a hydrophilic resin that would not swell the polysulphone). Then you should be able to section the membrane nicely without collapse of the structure.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Dear all, does anybody knows a protocol for the enzymatic localization of proteases at the ultrastructural level (dark precipitation products etc.)? Best regards, B.Laube Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit„tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
Dear Mr. Scharfeld,
we can send you a video animation by email. On this you can see one of our stepping motors working in vacuum. You can see how it works while a scratch test of a ceramic and metal sample is going on. If you think this stepper motor is appropriate for your stretch test, please give me a note.
Are you interested in this?
Best regards
Dr. Peter Marienhoff
___________________________ Dr. Peter Marienhoff VisiTec Microtechnik GmbH Karl-Marx-Str. 14 D-23936 Grevesmuehlen Germany
We are presently trying to measure the relative surface areas of small particles using SEM. Does anyone know of a technique that will enable us to do this? We are aware of the technique using stereo pairs and calculations that can be used to determine the relative height of a surface feature above or beneath the middle plane, but we are looking for a more straightforward way of estimating relative surface areas without having to calculate the height of each point in the micrograph. Any assistance would be appreciated, please respond directly to: farmer-at-cb.uga.edu
Thanks,
Mark A. Farmer Director, Ctr. Ultrastructural Research 151 Barrow Hall University of Georgia, Athens, GA 30602 (706)542-4080 Voice (706)542-4271 FAX farmer-at-cb.uga.edu http://www.uga.edu/caur
Hi Karen, We too have a Plasma Sciences coater, the CrC-100. The assets of Plasma Sciences were purchased by Torr International in early 1998. I received a letter from Torr Int'l in April 1998 stating that they could supply parts and service. Try contacting them at; 12 Columbus Street, New Windsor, NY, 12553. (914) 565-4027 or FAX (914) 561-7731. The president of the company is Masud Naraghi with email address; torr.intl-at-juno.com. The web-site is www.torr.com.
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
} -----Original Message----- } From: us312481 [SMTP:kszaruba-at-MMM.COM] } Sent: Monday, September 27, 1999 6:20 PM } To: MSA Listserve } Subject: Plasma Sciences Coater } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have a Plasma Sciences LVC-76 Sputter coater with Carbon attachment } that is having a mid-life crisis. It seems like some type of electrical } or wiring problem, or perhaps a faulty switch?? but it works } sporadically. (I am passing this on for a colleague, so am a bit fuzzy } on the details). At least once the coating discharged towards the } grounding wire inside the unit instead of evenly. Our internal } electronics service personnel have exhausted their expertise towards } repair. } } Any suggestions on where to turn to have this unit repaired? I } understand Plasma Sciences is no longer in existence. Alternatively, } any suggestions on a good model to purchase these days? Preferably a } company that will be around for a while (we've been through three } different sputter coaters in the last 8 years, all requiring service, at } least two of them from companies no longer in business.) } } Thanks for your help! } -- } Karen S. Zaruba kszaruba-at-mmm.com } 3M Center } St. Paul, MN 55144 } }
Hi,
Have you thought of confocal FT-IR? Renishaw and Instruments SA both have interesting instruments for this type of application (see "Focus on Microscopy", American Lab, July 1999 for a review of instrumentation at this year's PITTCON meeting). This combined technology is really great for just the purpose you have outlined.
Let me know how things go.
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 08:16 AM 9/24/99 -0500, Brian Reid wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
There are a number of specialized protocols used in the preparation of cartilage dependent on which matrix components one wishes to specifically visualize. Cells are best preserved using a quantity of Ruthenium Hexammine Trichloride (usually about 0.3% in glutaraldehyde, buffer and OsO4). This also tends to preserve cells somewhat better than fixation with no Ruthnium. However, RHT precipitates proteoglycans (which are otherwise leached out of the matrix) and this precipitate makes it difficult to see the morphology of collagen fibrils. If you are interested in observing collagen fibrils specifically, you may want to leave out the ruthenium and include 0.01% tannic acid in the primary fixative, which will stain the collagen fibrils.
We extend our infiltration time in Spurrs by 30 minutes each step for this dense tissue and try to keep our tissue blocks no larger than 0.5 mm on the narrowest edge.
Chondrocyte morphology by any chemical fixation will not be ideal. Cell shrinkage is a particular problem, and the cell borders will be seen to be very ruffled and retracted from the surrounding matrix. The best way to successfully preserve chondrocytes is by cryofixation (high pressure freezing) and freeze substitution.
I hope this helps!
Doug ---------------------- Douglas R. Keene Shriners Hospital Microscopy Facility 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
As of today 9-28-99 there are only 5 spaces left.
This is a great course for very little money.
The New York Microscopical Society
Bernard Friedman Memorial Workshop
Use of the Microscope
October 2, 9, 16, 23, 1999
A Basic Course on Light Microscopy which will cover the following topics:
Theory of microscopy Kohler Illumination Diffraction Theory Contrast Methods Polarized light Phase Contrast Interference Hoffman contrast Rheinberg Dark-field & oblique Illumination, etc.
The workshop will consist of Four Consecutive Saturdays of lectures and hands-on labs to cover the theoretical and practical aspects of microscopy.
The course instructors include:
Jan Hinsch of Leica, Inc.
Dennis O'Leary of Micro-Optical Methods
Mary McCann of McCann Imaging
John Reffner of Sensir Inc. and Current President of NYMS
Donald O'Leary, Curator and Awards Chairperson of NYMS
WHEN: October 2, 9,16, 23, 1999, from 10 A.M. to 4 P.M.
WHERE: New York Microscopical Society Facility 1244 McBride Avenue West Paterson, NJ. Phone (973)-812-8377
(Free parking, accessible by public transportation, Information on car pools and transportation will be provided.)
COST: $225 for N.Y.M.S. members, $245 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: Beginners and experienced users who wish to learn more about the proper use of a microscope.
For Further Information and To Register contact Donald O'Leary.
eMail: mailto:donoleary-at-worldnet.att.net
(201) 797-8849 Voice Phone Number
Limited to the First 12 Registrants.
Low Vacuum and Environmental Scanning Electron Microscopy, LV-ESEM '99
October 19-21, 1999, Chalmers University of Technology, Gothenburg, Sweden
This course will be given in close collaboration with four microscope manufacturers (Hitachi, Jeol, Leo, Philips/Fei) and suppliers of equipment for EDX, Cryo SEM and EBSP.
The aim of this 3-day intensive course is to give a theoretical background in the morning sessions and experimental insights in the afternoons. The lectures and the demonstrations will be given by application specialists from the different companies representad at the course and also by academic people working with LV- and ESEM. The demonstrations and lab classes will be carried out on equipment brought to Chalmers specifically for this course.
Detailed information about LV-ESEM '99, including course programme and registration form, is posted on: http://fy.chalmers.se/microscopy
Course organisers: Dr Lena Falk (lklfalk-at-fy.chalmers.se) and Dr Mats Halvarsson (mats.halvarsson-at-fy.chalmers.se)
Tom,
I am not sure how this would work, but there is a little tensile strength tester called the "Minimat" made by Rheometrics Scientific in Piscataway, NJ. Larry Green used to be productg manager, but I have not been in contact with them for a while. Suggest that you call the main number: 908-560-8550 and inquire further. This device fits on a light microscope stage and has a great control/measurement module which permits all sorts of stress-strain analysis.
Let me know how you make out.
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 12:06 PM 9/26/99 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
Some years ago a graduate student from another lab in the department came to me half hysterical, because she simply was not able to get label. I talked to her a long time, and then gave her a step by step protocol for GA and Osmium fixation (and etching, etc). I then helped her to make sure to minimize repeating the experiment. She came to me with the most remarkable micrographs. Beautiful label. Little or no background. Poster ready to do. After she left, I spent the day dumbfounded, literally stumbling over my own shoes and getting my labcoat caught in the door, because I could NOT do with the same protocol what she had done. And, I was the one that wrote the protocol. What was the difference? Only one. I had another antigen to locate than hers. What a lesson that was! If we can't stand the failures and variations that is due us when we do immuno, we better go to work in a bakery. There things are predictable. And, if we make a mistake, we can enjoy eating it!
Bye, Hildy Crowley
P.S. These "discussions" that let loose barrages of interest are wonderful. We learn so much!
THE NORTH CAROLINA SOCIETY FOR MICROSCOPY AND MICROBEAM ANALYSIS
present the
EIGHTEENTH ANNUAL SYMPOSIUM ON ADVANCES IN MICROSCOPY
"Nanoscopy: 'Microscopy' for the New Century?"
Coastline Convention Center, Wilmington, North Carolina
October 29 - 31, 1999
FOLLOW-UP ANNOUNCEMENT - WILMINGTON NOT FLOODED BY HURRICANE FLOYD! SYMPOSIUM IS ON SCHEDULE!
The Eighteenth Annual Symposium, sponsored by the North Carolina Society for Microscopy and Microbeam Analysis (NCSMMA), has been planned with a theme of "Nanoscopy: 'Microscopy' for the New Century?" Continuing with the tradition of the symposium, the guest lecturers are composed of both nationally and internationally distinguished scientists.
Speakers who have agreed to participate (so far) this year include:
* Stuart Lindsay (Arizona State Univ., Tempe) * David Wollman (NIST, Boulder) * Mike Isaacson (Cornell) * Richard Leapman (NIH) * Mike Kersker (JEOL) * Ken Taylor (Florida State) * Pat Calarco (UCSF) * Mike and Mary Reedy (Duke)
***** A FULL PROGRAM LISTING CAN BE FOUND ON OUR WEBSITE AT: http://152.3.167.174/NCA nnSymp99.html
* The meeting has several purposes, not the least of which is to draw attention of the scientific community to emerging developments in the practical and basic research aspects of exciting new fields, and to bring people together from diverse disciplines to discuss how innovative techniques will be relevant to the future direction of microscopy and microprobe analysis. In particular, this year, special emphasis will be placed on how recent advances in nanoscience and nanoengineering have resulted in new knowledge that has benefited microscopy in general and are having a significant impact in the biological and physical sciences. The symposium also offers an opportunity for interested participants including students to submit abstracts of related studies for poster display.
* 3 special workshops/tutorials will be offered at no additional charge to participants in the Symposium: (a) Cryo-preparation Techniques, (b) Atomic Force Microscopy (c) Digital Imaging Methods. These are practical, introductory sessions and no previous experience or knowledge is necessary.
Registration Fees, Hotel rates The $90 ($100 on site) per person and $50 for students ($60 on site). Registration fee includes: symposium attendance and materials, Saturday lunch, breaks, and Friday and Saturday evening meals. Additional Friday evening tickets are available for Adults - $20; Children 10 years of age and under - $10. Additional Saturday evening tickets are available for Adults - $20; Children 10 years of age and under - $10. There is a $15 fee for all cancellations. Coast Line Inn special rates $75/room/night (single or double). (Group No. 1226) Tel: 1 800 617-7732 or 1 910 763-2800
For questions or further information on Registration, please telephone Betty Gooch, Duke University Medical Center: (919) 286-0411 x 7266 or email: b.gooch-at-cellbio.duke edu
or ingram-at-rti.org
919 541-6598
We have a new SEM and confocal scope and we need to be able to track the usage of each of them. I was told that there is software that will do this. Can anyone offer help as to the name of the software and were I can get it? Thanks for your help.
Donna Wagahoff SIU School of Medicine PO Box 19627 Springfield, IL 62794-9627 217-782-0898 fax217-524-3227
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Karen S. Zaruba wrote: ============================================ We have a Plasma Sciences LVC-76 Sputter coater with Carbon attachment that is having a mid-life crisis. It seems like some type of electrical or wiring problem, or perhaps a faulty switch?? but it works sporadically. (I am passing this on for a colleague, so am a bit fuzzy on the details). At least once the coating discharged towards the grounding wire inside the unit instead of evenly. Our internal electronics service personnel have exhausted their expertise towards repair.
Any suggestions on where to turn to have this unit repaired? I understand Plasma Sciences is no longer in existence. Alternatively, any suggestions on a good model to purchase these days? Preferably a company that will be around for a while (we've been through three different sputter coaters in the last 8 years, all requiring service, at least two of them from companies no longer in business.)
Thanks for your help! -- Karen S. Zaruba kszaruba-at-mmm.com 3M Center St. Paul, MN 55144 ================================================ The "rights" to the manufacturing and servicing of the installed base of Plasma Sciences designed and produced LVC-76 units was purchased at the time of the bankruptcy several years ago of Plasma Sciences, Inc. by the following firm:
Torr International, Inc. www.torr.com
They are located in upstate New York.
The president of Torr (Dr. Masud Naraghi) is a real expert in plasma physics , and I have a lot of confidence in his design abilities and overall understanding of the physics of operation of these and other sputter coaters
Well, I just can't keep quiet any longer. Yes indeed, Laura, when you were cutting your teeth, I was cutting mine on an old RCA EMU-2D in 1968 at Georgetown University as the research assistant for Dr. George B. Chapman. And yes, to you too Kent. The same George B. Chapman AND the same RCA EMU-2D!! He is still there and so is the scope. As a matter of fact, the last time I spoke with him, he said that the was still managing to "squeeze a couple of respectable micrographs out of the ol' scope on occasion." And I don't doubt that he is. Even though we had a service contract for the scope, he did an awful lot of servicing it himself. When I left in 1976 he was still making his own apertures. They were quite a pair...that man and his scope!
After coming to UNC-Charlotte in '76 to be the caretaker of a Philips 301C there wasn't much left to learn. (That's an exaggeration, of course.) But WOW! External alignment. No more rubber hammers needed. Whoever was talking about contrast was quite right, too. I have not seen the contast that we got from that 2D since.
So today, I wrote Dr. Chapman a letter and enclosed copies of some of your comments. I think he will be pleased.
And John, if possible, I would love to have a copy of the article in the Daily News, April 27, 1986.
Sandra F. Zane, Electron Microscope Technician sfzane-at-email.uncc.edu Dept. of Biology, UNC-Charlotte Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
TEM JOB OPPORTUNITY
Assume responsibility for managing TEM services at LifeCell Corporation. LifeCell is a rapidly growing biotechnology company engaged in development and commercialization of tissue-derived products for tissue repair and transplantation. These products are based on LifeCell's unique tissue processing and preservation technology.
TEM facilities will be located in an entirely new building in Branchburg, an attractive north central New Jersey location.
In addition to overall management of TEM services, responsibilities include supervision of support personnel; preparation, evaluation and interpretation of complex tissue samples; preparation and delivery of written and oral technical reports of investigations; detailed record keeping and generation of creative solutions to solve technical problems. Required qualifications include minimum of BS degree, at least 3 years independent TEM experience with biological tissues, high level of organization, excellent oral/written communication skills and demonstrated ability for solid performance on several projects simultaneously in a team environment.
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Our EM specimens and records are considered as surgical specimens and are kept as per the following CAP guidelines: Blocks: 5 years Glass slides: 10 years Surgical reports: 10 years Wet tissue: 2 weeks after sign out
True CAP doesn't specify EM storage, but you can assume that if slides and reports (our reports include the prints) are held for 10 years that grids and negatives should be held as long.
We actually store all material forever, except for wet tissue which is discarded. We have warehouse space for older surgical, autopsy and cytology materials. The EM material is stored within the department. It is interesting that CAP only requires 5 year storage for paraffin blocks- probably because of the space needed to file and store them, not a problem with EM blocks.
Contact me if you need additional info about CAP inspections,etc. Becky Garrison Pathology Shands-Jackspnville 904-549-6072 becky.garrison-at-jax.ufl.edu -----Original Message----- } From: "drennie-at-UNMC.EDU"-at-Sparc5.Microscopy.Com [mailto:"drennie-at-UNMC.EDU"-at-Sparc5.Microscopy.Com] Sent: Friday, September 10, 1999 2:35 PM To: microscopy-at-Sparc5.Microscopy.Com
Good afternoon,
I have a question I have not been able to locate the answer to anywhere. I run a clinical EM lab here at the University of Nebraska and we have recently become associated with the College of Anatomic Pathologists (CAP) and I am having trouble locating any set regulations or even guidelines as to the duration we should retain records, as well as specimen blocks, thick section slides and EM photos. Does anyone have any info to get me headed in the right direction?
Thank you,
Doug Rennie Coodinator-EM Facility University of Nebraska Medical Center Omaha, Nebraska.
Greetings:
I am helping someone who wishes to compare the staining intensity of a tissue area across several slides and treatments. The intensity goes from nothing (control) to something pretty dark. He would like to rank the staining intensity using photographs or some other appropriate technique. Some of the differences appear to be subtle.
I don't know the best way to do this, though it seems like a reasonable request. I mostly thought of problems, ie section thickness, non-linearity in the film response, variation in exposure, compensation by his eye, etc. etc.
Am I overly cautious? Is there a good way to do this kind of thing? Any advice?
If you have some ideas or want more details about his work and objectives, please contact me.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
----- Original Message ----- } From: Karl E. Garsha {keg-at-csd.uwm.edu} To: Jon Krupp {jmkrupp-at-cats.ucsc.edu} Sent: Tuesday, September 28, 1999 7:04 PM
Hi, Everyone
I'm shortly going to buy a new EDS detector. I'd quite like the performance advantages of a 1-atmosphere UTW, but I'm concerned about the durability and longevity compared with standard Be windows.
Anyone out there got any relevant experience/views?
Either to the list, or directly to me, please.
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Thanks to everybody for their many helpful comments on the problem of streaking of the film during developing of SO163. (so many, in fact, that I cannot reply to everyone individually) The streaks look like 0.5-1 cm bands of alternating low/high density, vertically with respect to the developing tank.
I did not mention that indeed we do not yet have nitrogen burst installed, so it is quite possible, as many of you suggested, that lack of agitation is the culprit. (I will find out soon, hopefully.) As one pointed out, manual agitation may even be too "good" in the sense of being too frequent and too regular (in the same direction). Indeed, the instructions from Kodak do recommend the use of "random movements" for manual agitation.
Most seem to agree that the temperature, especially of the final wash is not that important, although some mention the temperature of the rinse between developer and fix as a potential problem. Of course, this doesn't have to be running water, so it's easier to control.
Another possibility that was mentioned is that the batch of film itself may be faulty. Others have also reported problem with specific batches of SO163. And who knows what it went through during shipping to Singapore.
In the previous labs that I worked there was always an expert technician who must have solved these problems long ago! Lacking this, the advice from this bulletin board has been extremely useful. Thanks!
terje
------------------------------------ Dr. Terje Dokland Senior Scientist Institute of Molecular Agrobiology 1 Research Link The National University of Singapore Singapore 117604 Phone: 65-872 7405 Fax: 65-872 7007 E-mail: dokland-at-ima.org.sg http://lab.ima.org.sg/lab3052/terje ------------------------------------
Hi,
Although there are some people who try to do this with scanners, etc., the proper way to do it is to use a laser densitometer. A scanner (except the high-end devices) do not have a sufficient linear response and whide enough dynamic range to do this kind of densitiometry measurements.
Sincerely yours,
Peter Van Osta, MD
Senior Scientist Medical Image Analysis Biological Imaging Laboratory Life Sciences Department I - 6065 Janssen Research Foundation Turnhoutseweg 30 B-2340 Beerse Belgium Europe
-----Original Message----- } From: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Tuesday, September 28, 1999 11:10 PM To: Microscopy-at-sparc5.microscopy.com
Greetings:
I am helping someone who wishes to compare the staining intensity of a tissue area across several slides and treatments. The intensity goes from nothing (control) to something pretty dark. He would like to rank the staining intensity using photographs or some other appropriate technique. Some of the differences appear to be subtle.
I don't know the best way to do this, though it seems like a reasonable request. I mostly thought of problems, ie section thickness, non-linearity in the film response, variation in exposure, compensation by his eye, etc. etc.
Am I overly cautious? Is there a good way to do this kind of thing? Any advice?
If you have some ideas or want more details about his work and objectives, please contact me.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Hi Ritchie,
We have had (Oxford Instruments) atmospheric thin window (ATW) and Super ATW detectors on SEM and TEMs for several years now. We are very happy with their performance and durability.
The differences that I have noted are that I am very careful to let the column up to atmospheric pressure or 0.1bar above atmospheric at the most. I always ensure that the column is fully up to pressure before breaking the column and I remove an aperture mechanism before removing the detector to ensure that I don't get a sudden change in pressure around the detector. Apart from that no extra precautions and no worries.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, Everyone } } I'm shortly going to buy a new EDS detector. } I'd quite like the performance advantages of a 1-atmosphere UTW, but } I'm concerned about the durability and longevity compared with } standard Be windows. } } Anyone out there got any relevant experience/views? } } Either to the list, or directly to me, please. } } thanks } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Hi all! The electronic addresses are necessary to me, or it is simple the names of the manufacturers electronic microscopes.
# Barnaul, Russia # Alexey V. Kalinin # Altay State University # avk-at-ic.dcn-asu.ru # Internet Center # http://www.dcn-asu.ru/~avk
Ritchie, Good to see your post. Hope all is well in Auckland. We have three systems with an UTW. 2 of them are NORAN Pioneer detectors, and the other is Oxford (Link Pentafet). They are all three great detectors, and it is the wisest move we ever made when we opened that door and went to the thin window detector. We have no regrets at all! One of the detectors (now, our oldest Pioneer) is on an 840A, as I assume you will be doing. We absolutely love that system. It has great durability/reliability (we've had it ~8 years), has never been back to the shop, and gives us performance above our expectations. Durability and longevity were a problem with the very first UTW that we had on a TEM back in the late '80s, but this is certainly not a concern of ours with these newer versions of the UTW. Go for it! Brad Huggins BPAmoco, Naperville, IL
} ---------- } From: Ritchie Sims[SMTP:r.sims-at-auckland.ac.nz] } Sent: Wednesday, September 29, 1999 9:48 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ultrathin Window vs Be } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, Everyone } } I'm shortly going to buy a new EDS detector. } I'd quite like the performance advantages of a 1-atmosphere UTW, but } I'm concerned about the durability and longevity compared with } standard Be windows. } } Anyone out there got any relevant experience/views? } } Either to the list, or directly to me, please. } } thanks } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
We have three ATW detectors with an aggregate life of about 15 years. Two of the windows have never failed (knock on wood!), the third has twice been damaged by clumsy stage manipulation in the SEM. Obviously this would have happened even if the window was Be, but in that case we would have had tiny fragments of highly toxic beryllium floating around the microscope chamber, instead of relatively benign polymer.
True, a week or so after one of the repairs, the new window did fail spontaneously, but almost certainly because of a nascent flaw - the manufacturer must have thought so, because they fixed it again free of charge.
Hope this helps,
Tony Gaarratt-Reed.
At 02:48 PM 09/29/1999 GMT+1200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am hoping that someone has a better method for this procedure. I am preparing small pieces of mouse intestine for SEM. We are looking at the villi and small Peyer's patches between villi. Perfusion is not an option so we excise the pieces and drop them into 2% paraformaldehye and 2% glutaraldehyde in cacodylate buffer as quickly as possible. I fix for several hours, wash with buffer and postfix with osmium. I have tried both CPD and airdrying after using hexamethyldisilizane. But when I look at the villi, it sometimes appears that there are "cracks" on the surface that separate bundles of villi, giving the intestine surface a crackled appearance. Is there any way to prevent this all the time? It's not consistent but it is frequent. Any help would be appreciated!
Thanks!
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
Jim,
I agree with most of what you have said about the early mechanical advantages of Siemens and Philips over RCA, but I don't (and didn't) arrive at the same conclusion that using RCA was a big mistake. I did electron microscopy for my biological PhD thesis at Harvard about 1956-58 on an RCA EMU-2D. During my postdoc with Don Fawcett (at Cornell Med and then Harvard Med) in 1958-61 we used mostly RCA EMU-3. A Siemens IA was available to us during part of that time, but wasn't used much. As you pointed out, the long focal length of the RCA instruments was great for contrast, but less optimal for resolution. But why worry about only getting 7A resolution (instead of 4-5A) when the smallest structure of appreciable biological interest visible in EM sections was about 25A (sublayers of biological membranes). Was there any advantage in seeing the stain particles more sharply? On the other hand, contrast was extremely important, and contributed to some of the high quality EM produced by Fawcett and his associates.
When I left for my first faculty position (Anatomy Dept at Stanford Med, 1961-71), I had grant money for an EM, and picked an RCA EMU-3F. I never regretted that choice. The high contrast of that instrument was invaluable, especially when (beginning in 1966) I was developing a method and device for cutting ultrathin frozen sections (which were inherently low in contrast).
Although my later instruments were a Philips 300, JEOL 100B, and then Philips 201, Philips 400 and Philips CM10, I still look back on the RCA as a great instrument, despite its various inconveniences.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Anatomy and Cell Biology University of Michigan Medical School Ann Arbor, MI 48109-0616 Tel (work) 763-1287, Fax (work) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} Hello List Readers: } Laura Rhoads suggested that forced use of THAT instrument would be a } suitable penalty for those committing "crimes" such as broadcasting } "dogma". I concured, adding "I would not want to offend anybody, but } I've been in purgatory for two years already. Circa 1970 that instrument } was the worst money could buy in the Western World." } } Several readers have responded making positive points about that } instruments. I agree with some of these because nothing in this world is } entirely bad - even a broken clock shows the right time, twice a day. } } Yes, that instrument looked good, it had a big column and would make a } fine statue in someone's garden (so Laura suggested, personal } communications). I would love one, it would be the crowing glory to the } banks of the Ross River. } } Yes, the image had good contrast because it was a medium resolution } instrument (I guess about 0.8nm). A longer working distance objective } results in higher contrast but lower resolution. Other manufacturers } offer high contrast objectives, but most instrument purchasers ask for } higher resolution. } } Yes, RCA's are historical instrument and the first commercially produced } American TEM. Though I believe that Siemens produced the first commercial } TEM in the mid 30th. } } Yes, for some people this was their first electron microscope and it had } to be impressive to us who grew up before computer whizz-bangery. } Unfortunately, it was my fourth TEM. My previous loves had been a } mid-60th Siemens Elmiskop, a Philips 100C (the one with a near } horizontal column and transmission viewing screen) and a Zeiss 9C. All } of these were rather better suited to productive work. } } Yes, the electronics were reliable, but the vacuum gauging was } insufficient. Since it also had no vacuum locks and blanking provisions } were poor, vacuum trouble-shooting was very difficult. } } Yes, it was reasonably easy to operate and was great for forced coffee } breaks. No specimen, no gun, nor camera vacuum locks made for lots and } lots of pumping times. } } Yes, the camera had two options, either three single plates could be } inserted or one long plate taking five images on one plate. Bit of a } pain to share negatives with another operator. Absolutely awful for } taking lots of images and changing specimens. The pumping cycle was slow. } } The complete lack of vacuum locks and the poor film options made the } EMU-4 a "hysterical instrument". Such features were not rocket science } even in those days. More features . . . } } Changing the filament was a painful operation. I think the basic } alignment after filament change required three complete column pumping } cycles, lots of time for getting other jobs done during those } operations. I could not align that scope in under two hours. I had } inherited a service contract and the service men could do no better. } } Alignment screws were difficult to adjust accurately in those days on } most TEMs. I learned from the service man that the final touch was best } achieved by tapping the column with a rubber mallet - which was not part } of the service kit. } } Filament life averaged about 12 hours, with the alignment procedure } eating up a good part thereof. Short filament life may not have been an } inherent problem in the design. This may have been due to poor vacuum, } but there was no good way of isolating vacuum parts, getting a reliable } vacuum reading and finding a possible leak. } } } Why did RCA sell the EMU-4 to Forgflow, who then quickly discontinued } that instrument? } Because the column design was 10 years too late. By 1972 I had a Philips } EM300 - now that was progress. Philips at last had eclipsed the } Elmiskops, which had been the top brand throughout the 50th and 60th. } } Incidentally, Siemens closed their EM division about '78. In Karlsruhe in } October '77 I had admired Siemens' latest "Wunderkind", it turned out be } the Divisions death knell. It was a FESEM based somewhat on the research } by Crewe (Chicago ?) Nice instrument, beautiful column, but Vacuum } Generator of the UK had build a better performer at almost half the } price. . . . the rest is history. } } Disclaimer: Opinions expressed are mine they are based on fragile } memories. No dogma is implied or intended. Feel free to purchase an } EMU-4, if you can find one. Even Phantom comics of that period fetch } increasing prices and modern instruments depreciate rapidly. Don't } condemn me for another two years on THAT instrument please. PST does not } supply the EMU-4 or similar. } 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 } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com
} X-Lotus-FromDomain: ECCI } Date: Wed, 29 Sep 1999 10:41:17 -0400 } Subject: Re: more fond RCA memories } Content-Disposition: inline } To: "Sandra F. Zane" {sfzane-at-email.uncc.edu} } From: dgibbon-at-ecc.com } Indirectly from Don Gibbon via S. Zane. Here are his own fond RCA memories. }
} } Well, I've got a few myself! } } } } When I got back from military service to Rice University (nee "Institute") in } 1961, someone had given the Geology Department a Shimadzu TEM, the only one ever } to come into this country, as far as I know. No one else was interested, so for } three years I taught myself how to run it, using its quasi-English primitive } manual and Hall's Intro to EM text (1953) for guidance. As an example of its } crudeness, I made my own filaments from a coil of tungsten wire. First I bent } } the wire into a V-shape by pressing it down over a single-edged razor blade. } Holding the crimped wire in one hand, I rubbed a flat spot onto the peak of the } "V" on the unglazed area on the bottom of a china cup! This was the level of } spohistication I was at, when one day I was ushered into the dark and quiet } chambers of the TEM lab in the nearby biology building, where sat an EMU3A. I } couldn't believe it! But I was only allowed a brief glimpse of what life might } be like. Ultimately I was sent up to Drexel for a two-week short course in 1964, } where people like Dave Ballard of NBS were also learning the arts on a EMU3B, I } believe. I've never forgotten the feeling of luxury as I sat and spun the silky } stage controls and watched the sample whizzing by on the screen! We visited the } RCA plant nearby in Camden, where my major memory is of the smell of soup from } the Campbell's cannery down the street! } } Eventually I took the TEM job at the Mineral Constitution Lab at Penn State, } where in 1964 they still had an EMU 2D in working condititon (as well as a } Hitachi 7A). The EMU 2D was a lot like an old 1930 Citroen I used to have... } with a gravity-feed fuel system: the gas flowed down of its own accord from a } tank behind the dash board! The EMU 2D was so simple that you actually had to be } careful not to pull parts of the column out by hand while it was under vacuum! } One thing about it: you really understood the physics of the microscopy process } when you used those instruments. There were no black boxes! } } } In those days, though, with the exception of the rare Siemens or Phillips, the } RCA was the top of the line, a really classy instrument. } } } } Donald L. Gibbon } } Calgon Corporation } } Materials Microcharacterization } } } } } }
Are there any particular pitfalls involved with the fixation of mouse embyos (ME9-17) for immunoEM-problems with osmolarity, for instance? I realize fixation is dependent on factors such as preservation of the desired antigen but I'm wondering about the osmolarity in particular. My limited previous experience with embryonic material (freshwater fish) was that it is, but I've never dealt with mammalian tissue of this age. Thanks Grace
I like to know if anyone has vibratome in Boston area and be able to give a demo if it's possoble. Thanks.
One remark regarding MPEG (see message below):
MPEG (the compression used by the card mentioned) is a lossy compression just like JPEG. This is usually not an issue if all you want to do is record a sequence of images for later viewing. However, if you want to do any kind of image processing, you might run into trouble as the compression leads to loss of information.
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 ===================================
-----Original Message----- } From: Shane Collins [mailto:kshanec-at-gte.net] Sent: Sunday, September 26, 1999 5:59 PM To: Michael Bode
Nice discussion and helpful hints about LR White/LR Gold.
The terms catalyst and accelerator have been used often in the reply. I am still unclear if the benzoyl peroxide is something that needs to be added to each batch of resin... or only if a cold cure method is used...or if there is another compound that is added as an accelerator for the cold cure. If so what is it? Are these terms being used interchangably?
FYI ....the blocks did polymerize at 50 C. after they were reinfiltrated with resin containing a complete formulation from the manufacturer. I will be trimming them carefully with glass knife and boat as suggested and will view un- ICC-stained grids this week. The investigator wants to do the ICC in his lab, so I will just be cutting grids and we will see what happens. Thanks for all your help !! Linda Fox lfox1-at-wpo.it.luc.edu
Is anyone out there using the DSG1 Plus digital scan generator for a JEOL SEM?? We just had a new unit installed on out JSM 840A running Windows98. The documentation is sparse to say the least, one laminated sheet, and we are muddling through all of the menu items.
Some of the options are not intuitive, such as setting micron bar scale factors. Options such as, I think pixel resolution, i.e.320x240, 2560x1920,1280-} 640 coupled with times i.e. 10,20,40,80...320 seconds. It is hard to determine which combination of settings to use, and what are the advantages/draw backs of each.
I gather that this is a new system to JEOL and they are learning it along with us...but right now time is tight and great instructions would go a LONG way. Can anyone help with this particular system?? Thanks, Linda Fox lfox1-at-wpo.it.luc.edu
Dear list members:
For studying polymer blends by using SEM, only information on surface of a sample can be obtained. Because the surface can not always be the plane of the center of each dispersed phase particle, the statistic result of dispersed phase particle size attained from SEM micrograph may be less than the true value.
By using image analysis we can easily get the spherical particle diameter distribution and usually it shows a log-normal distribution. However, it is not the "true" distribution. Does anyone has some advice on how to convert this measued distribution to the "true" one?
Another question is when the particle is ellipsoid, how can I find its proper size from image analysis of SEM photo?
Thank you in advance for your help.
Dr. Zeng Jijun Visiting Scholar of Toray
Home Address: Nakatogari 734-2-5 Nagaizumi-cho, Sunto-gun Shizuoka, 411-0942 Japan
Personal Homepage: http://www.geocities.com/ResearchTriangle/Forum/1786
I have read with interest the various responses to this question. I have one related question in this regard to which I would appreciate expert opinion.
Choice of UTW is usually made with respect to the need for light element analysis. One disadvantage having an UTW detector is that it precludes the use of a heating holder for in-situ studies. Is there a way to overcome this limitation i.e. be able to do in-situ phase transformation studies in a TEM with an UTW detector fitted? If chemical analysis is not required is it safe to retract the detector from the column (I believe it is possible to change the position of the detector by a few centimetres) and use the heating holder? If yes, what distance is considered safe?
Thanks. ---- Divakar R PMS, MCG, IGCAR ----
-----Original Message----- } From: Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] Sent: Wednesday, September 29, 1999 9:47 AM To: Microscopy-at-sparc5.microscopy.com
Hi, Everyone
I'm shortly going to buy a new EDS detector. I'd quite like the performance advantages of a 1-atmosphere UTW, but I'm concerned about the durability and longevity compared with standard Be windows.
Anyone out there got any relevant experience/views?
Either to the list, or directly to me, please.
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Hi again
So far two replies to my earlier posting regarding durability of UTWs, both positive about them, anybody got any experience to the contrary?
Further query:-
Like many, I haven't yet gotten around to putting in a foreline trap (next week, for sure), so my EDS detector window (Be) needs to have the oil cleaned off it every six months or so, otherwise Na takes a slow dive. I do this by gently dribbling Freon over the window.
Can this be done with impunity with an UTW?
Perhaps a manufacturer might like to reply.
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Hi Richie,
I would like to follow up your comment about the foreline trap and the constant need to clean the Be detector window. Why do you want to get an ATW detector? - presumably for the ligher element analysis. If this is so then I would strongly advise you to get your vacuum system sorted before you install the detector. It is my experience when using windowless, UTW and ATW detectors that vacuum cleanliness is the just about the most important factor in light element analysis. The buildup of hydrocarbon contamination ruins any light element analysis. Detectors also need conditioning more frequently (every 3 to 6 months depending on your needs) to remove the ice buildup.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 again } } So far two replies to my earlier posting regarding durability of } UTWs, both positive about them, anybody got any experience to the } contrary? } } Further query:- } } Like many, I haven't yet gotten around to putting in a foreline trap } (next week, for sure), so my EDS detector window (Be) needs to have } the oil cleaned off it every six months or so, otherwise Na takes a } slow dive. } I do this by gently dribbling Freon over the window. } } Can this be done with impunity with an UTW? } } Perhaps a manufacturer might like to reply. } } thanks } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Linda - } } Nice discussion and helpful hints about LR White/LR Gold. } } The terms catalyst and accelerator have been used often in the reply.
These terms are sometimes used in a less-than-rigorous manner; I'll use the terminology from Glauert & Lewis, "Biological specimen preparation for TEM", 1998. Dibenzoyl peroxide is an "initiator", required for the polymerization of ALL LR White. Since its addtion makes it possible to polymerise the resin with heat, some suppliers ship the bottle and the DBP separately, to avoid premature hardening during shipment. Check the label on receipt, and add a dated comment if you mix in the DBP yourself.
} I am still unclear if the benzoyl peroxide is something that needs to } be added to each batch of resin... or only if a cold cure method is } used...or if there is another compound that is added as an accelerator } for the cold cure. If so what is it?
There is a separate liquid "accelerator" for "cold" cure; its composition is proprietary. Please realise that this polymerization isn't cold; it's quite exothermic.
CCCCCaroline
Caroline Schooley Project MICRO 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 work in a hospital based diagnostic EM service. We keep all autopsy and surgical materials forever except wet tissue which is disposed of on a periodic basis.
True, there aren't CAP guidelines for EM specifically but EM is considered a surgical procedure and falls under the following surgical guidelines (as per our 1997 CAP survey): Blocks: 5 years Glass slides: 10 years Reports: 10 years Wet tissue: Surgical: 2 weeks after sign out Autopsy: 3 months
So CAP doesn't address grids, prints and negatives. We consider the EM blocks and negatives material which will be stored forever because the work can be reproduced from them.
I do discard wet tissue which is not processed. If you have other questions, contact me.
Hi evrybody In order to plan a budget for a future FEG I would like to know the following point concerning the commercial thermal FEG.
-type of instrument -How long is the life time? -What is necessary to exchange, tip only or gun set? -How much it cost? -How long it takes to replace -which garantee on lifetime? -comments?
Please answer directly to author. Thank you very much for your help. Serge NITSCHE.
Hi, Linda,
I had osmium fixed specimens polymerized in LR White resin during infiltration in the past. I came to conclusion that the resin was old.
My supplier would sell me uncatalysed LRW. I mixed the supplied catalyst, benzoyl peroxide, with LRW just before using it. I divided it into several small bottles and store in the frig to improve shelf-life. Each small bottle is consumed before another is opened and I do not have premature polymerization any more.
Accelerator is required for cold curing the LRW mixed with bp.
} } } "Linda Fox" {LFOX1-at-wpo.it.luc.edu} 09/29 3:26 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Nice discussion and helpful hints about LR White/LR Gold.
The terms catalyst and accelerator have been used often in the reply. I am still unclear if the benzoyl peroxide is something that needs to be added to each batch of resin... or only if a cold cure method is used...or if there is another compound that is added as an accelerator for the cold cure. If so what is it? Are these terms being used interchangably?
FYI ....the blocks did polymerize at 50 C. after they were reinfiltrated with resin containing a complete formulation from the manufacturer. I will be trimming them carefully with glass knife and boat as suggested and will view un- ICC-stained grids this week. The investigator wants to do the ICC in his lab, so I will just be cutting grids and we will see what happens. Thanks for all your help !! Linda Fox lfox1-at-wpo.it.luc.edu
Attn all RCA-o-philes,
With regard to Jim Darley's citing of the High Crimes in the Diet of Dogma Court Record... } } Hello List Readers: } Laura Rhoads suggested that forced use of THAT instrument would be a suitable } penalty for those committing "crimes" such as broadcasting "dogma". I } concured, } adding "I would not want to offend anybody, but I've been in purgatory for } two } years already. Circa 1970 that instrument was the worst money could buy in } the } Western World." } } Yes, that instrument looked good, it had a big column and would make a fine } statue in someone's garden (so Laura suggested, personal communications). I } would love one, it would be the crowing glory to the banks of the Ross River.
Jim- You've got it all wrong: I said lawn ornament, not garden. If I put an EMU-4 column and requisite support pedestal in my garden there would be no room for my radishes... Actually, someone has already offered me an RCA EMT for this purpose, but that model isn't nearly massive enough for what I intend (I think it's complete so if anyone wants this artifact say, for a home workshop restoration project, contact me off-list).
Actually, the subject of RCA-for-pillory has brought up some rather interesting interpretations regarding the application of RCA EO equipment. Since I had yet to enter kindergarten by the time the last EMU-4 rolled off the assembly line in 1969 I never had the opportunity many others seem to have had cutting their teeth (I was cutting my own) on this (at the time) cutting edge technology. Missing this equipment in its prime, I feel somehow cheated, and as a result will have to rely on the experiences of those who did. What I find really interesting is the disparate variety of opinions as to service and operation. Could the EMU-4 Jim used at the World Expo have been a made on a Monday and Chuck Garber's, for example, a Wednesday? Or, did the RCA engineers install some extra parts they found in Hangar 18 from the Roswell, NM Crash Site that Jim's didn't get? Since the SPI rig is still running perhaps the operators can comment on their filament life and if it exceeds 12 hours? In order to solve the poor vacuum pumping problem (and to avoid forced coffee breaks) has this unit's vacuum system been retrofitted with a Balzers turbo pump perhaps? Are the alignment problems vividly recalled simply the result of using the wrong size mallet maybe?
All these issues are quite intriguing. If anyone does look into the history of RCA then maybe other manufacturers of EO equipment, such as Siemens, would be in order as well? And who was Vacuum Generator of the UK? Those who forget the past are doomed to repeat its failures. It seems that the technical information is available, and MSA has a tremendous body of institutional memory. It would be a shame to have this collective wisdom disappear.
For that matter, when I arrived at my latest job I discovered a complete Philips 75C hidden in the corner of the basement, with original warranty card (never filled out) which I am told still operates...Where will I ever get parts?...
************************************************ If the iron dice must roll, may God help us all...
Theobald von Bethmann-Hollweg German chancellor, August 1, 1914
************************************************ Laura Rhoads Biology Department SUNY Potsdam Potsdam, NY 13676 315-267-2260 315-267-3170 fax
I have a Philips 201 TEM with some major minor problems. I need to replace some small parts to fix the specimen stage.
Most essential: a stereoblock (a cube about 2cm on side, tube hollowed out inside, small projecting knobby thing from one side). It is part # 5322-466-80154 in the Philips 1974 manual. I also need a leaf spring and pin that fits into this - # 5322-492-6061.
I would also like to buy (some) replacement clamping cones for the specimen holder and an new objective diaphram holder tip, but these are less critical at the moment.
Does anyone have or know where I can acquire such parts? If anyone is trying to get rid of a defunct Philips 201 TEM...? According to the manual parts for the Philips 300 are the same.
Thanks! g
Many thanks indeed to all who responded to the above distress call. My colleague Janet says that she will try all the suggestions and report results. She finds Mike Wombwell's suggestion particularly promising. Chris Smith, Plant Path EM Unit, IACR-Rothamsted, UK.
Colleagues
I have discovered that the main database file became corrupted a few days ago. As a result many of you (about 2000+) have not received any Listserver mail for the last few days.
Unfortunately I'm out of the country at the moment and will not be able to do a full restore until Monday when I return home.
I have been able to restore an older database which I have here on my laptop, which will get most people back on-line (as well as a few that have UNSUBSCRIBED ... sorry). Please bear with this for a bit until I can get things back to normal.
Just for the record the problem, (which I probably caused) happened on Mon, 27 Sep 1999 17:20 -0500 CST. A large portion of the database file was deleted. oops... Nestor screwed up!
This means that most of you will have had a quiet few days. Even the archive records were lost for that 3 day period. I 've been out of the country for 2 weeks and have not been monitoring the Listserver mail closely so I did not catch the problem until a host of people started sending me Email asking why they haven't seen anything for the last 2-3 days . My thanks to everyone who sent me an Email that something was amiss.
If any of you that were still receiving Listserver mail during 9/27-9/30 period (I obviously did not) and have happen to have saved all the messages you received during that drop me a line. I'd like to try to organize a restoration of as much to the archive as possible.....
Also my apologies to any of you that have unsubscribed and are now receiving this mail. Please send a new Unsubscribe message to
Listserver-at-msa.microscopy.com
I'll be able to do a more complete restore of the database in a few days when I return home.
I have used PIPS ion beam thinner for a month or so. Vacuum level became bad and I could not turn on the gun. When I opened up the chamber, I found that metal parts were contaminated severely. It seemed that it was time to clean up the parts. I tried to clean them with tissue paper and alcohol. They were hardly removed. Especially when the metal surface had a machining marks such as concentric circular grooves, it was more difficult. Some parts I could take out of chamber and clean. The other parts, I could not take out because they were stationary. I have to clean them in a state of being equipped. I am afraid to use organic solvent which would remain in the chamber and make the vacuum worse. My question is how I can clean the contamination easily. Could any of you give me help out of your good experiences? Help from Gatan expert would also be appreciated.
Jondo Yun Center for Instrumental Analysis Kyungnam University 449 Weolyeong-dong, Masan, 631-701, Korea 82-551-249-2697 (tel) 82-551-248-5033 (fax) jdyun-at-hanma.kyungnam.ac.kr
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