The software for a conversion of AN10000 to Windows has already been written by a Toronto based company called Mektech. They also make a hardware to interface the AN10000 pulse processor to a PC. You can find them at mektech-at-visionol.net The Microscopy Imaging Laboratory at the University of Toronto Dept. of Medicine. Has purchased the interface for the AN10000 to PC and the software from Mektech. The software is easy to operate and we have found it to be very reliable. Mektech has allowed us to upgrade our system at a fraction of the cost of that of a new system.
Battista Calvieri Manager of Microscopy Imaging Labarotory University of Toronto
Not even the suggested sophistication of a "halon" fire protection system will render your EMs immune from the dreaded "water shower" experience. Overnight flooding of the lab floor above us caused water to find its way to (and then through) the smoke/ thermal detectors that we had smugly positioned directly over the instrument for our halon gas quench system, cascading water onto a powered TEM/STEM/EDS facility. By the time I was called in, the TEM was lit up like a Xmas tree, accompanied by the delicate aroma only smoldering electronics can generate.
Unfortunately, similar water showers in the adjacent SEM room had missed the console, and the instrument was too heavy to push under the water stream (we were trying to get a new SEM at the time!).
Note: Our fire detectors have since been relocated, and the "halon" has been replaced (by Govt decree) with environmentally more friendly (less unfriendly?) FM200.
Eric Bradley BHP Steel Port Kembla NSW Australia bradley.eric.eg-at-bhp.com.au
---------------------------------------
Some months ago a gate valve went in the cooling pipes for our building, which created a spectacular waterfall in the corridor. Since an insurance claim is the easiest way of getting new equipment here, we were quite hopeful, but like Eric Bradley, we found that three strong people were unable to push either our 17 year old TEM, 25 year old Auger kit, or even the little 18 year old SEM under the water.
I have a problem about cell number count on agrregated neurons with an enzyme treatment. I want to check a viability after the enzyme treatment. It is usually available to count a cell number under phase contrast microscopy, but light was so reflected that each neuron could not be identified. I would, therfore, try to apply a fluorescence dye,for example, Hoechst33342, that bind to a minor grove of DNA and is heared to be applicable to a native cell. But I wonder it will work well. I would like to get more detailed information about image analyze of cell number under a microscopy. Thanks a lot.
Masahito Morita Laboratory of Compararive physiology Faculty of Science Osaka University
} I am trying to compile a list of what type of disposable gloves should be } used with what chemicals found in our EM unit. [snip] } } Organic solvents: } } acetone: butyl? or latex } ethanol: latex } methanol: latex } chloroform: (PVA if avail) or nitrile (double glove) } [snip]
Dear Richard, I'd be very surprized if latex gloves would hold up to acetone; when I've exposed them, they get tacky. Polyethylene gloves should hold up to the organic solvents you listed (and many others). Besides, they smell better than latex gloves. Yours, Bill Tivol
Richard, and others interested;
We use 100% Nitrile gloves as they stand up to all the chemicals we use in the EM lab, We use the product available from Best Manufacturing Company, Menlo, GA 800-241-0323. I'm sure there are others just as suitable.
Disclaimer: We have no financial or other interest in the company.
But I've been on the road for awhile and hence did not have time to catch up on things. Here's the info that a few people have asked for as I did not see anyone else post an answer.
The MSA/MAS Spectrum File Format Information & Examples can be downloaded from the following FTP Site.
Host: ftp.amc.anl.gov UserID: anonymous Password: Your Email Address
Everything is in the EMMFF ( "E"MSA MAS File Format) directory, including examples and documentation.
Version 1.0 is the current revision level. We (The MSA/MAS Standards Committee) have not had a need to update it beyond that point.
I have not gotten around to creating a WWW page for this. If there is enough interest I will do it. However if you have access to the WWW you can get it via FTP. It is also routinely available in the Computer Workshop at the annual Microscopy & Microanalysis Meeting . ( see http://www.msa.microscopy.com for this years meeting information )
Cheers... Nestor Your Friendly Neighborhood SysOp
Here is the Abstract File for that directory.
-------- Title :EMMFF V. 1.0 Keywords :XEDS,EELS,AES,WDS,CLS,GAM,XRF,PES Computer :IBM, MAC, DEC Operating System :ALL Programming Language :Fortran 77 Hardware Requirements :None Author(s) :EMSA/MAS TASK FORCE Ray Egerton ,Charles E. Fiori ,John A. Hunt, Mike S. Isaacson,Earl J. Kirkland ,Nestor J. Zaluzec Correspondence Address :R.F. EGERTON-CHAIRMAN University of Alberta Dept. of Physics Edmonton, Alberta, Canada, T6G2J1 Abstract:
A simple format for the exchange of digital spectral data is presented, and proposed as an EMSA/MAS standard. This format is readable by both humans and computers and is suitable for transmission through various electronic networks (BITNET, ARPANET), the phone system (with modems) or on physical computer storage devices (such as floppy disks). The format is not tied to any one computer, programming language or computer operating system. The adoption of a standard format would enable different laboratories to freely exchange spectral data, and would help to standarize data analysis software. If equipment manufacturers were to support a common format, the microscopy and microanalysis community would avoid duplicated effort in writing data-analysis software. This version of EMSAMASFF contains two subroutines which read and write spectral data files Version 1.0 data format. The data are stored as simple ASCII characters at a user defined number of columns per line for the length of the data file. The spectral data is preceeded by a series of header lines, which tell the user about the parameters of the spectrum. The header lines are identified by the first character in the line being the symbol (#) followed by a descriptor and if appropriate its units. An example of a data file format can be found in the EMSAMASFF.DOC file. ------------------------------------------------------------------------------
You never know what you will find in your e-mail "in" basket. If any of you know the derivation of the word "Wehnelt", please respond to this gentleman directly at:
} The subscriber is obviously a relative of this "wehnelt" you mention. I } belong to the Brazilian side of the family, hence the different graphy of } the name. Can you be kind enough to give more info on this man? I knoe there } was an Arthur Waehneldt who dedicated himself to physics. Are we } talking about the same person? Pls. send all the details you may have. I`m } trying to trace member oof the family ald your cooperation will be very } wellcome. Thanking you in advance, I remain. } Gustavo } gustavo waehneldt } guswae-at-tricom.net ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Good morning, I would like to thank everyone who so helpfully provided information on servicing the stereo pod on my Ultracut:
Lou Ann Miller, Julian Smith III, Normand Laurier, Allan Mitchell, Kevin Hlacrow, Sara Miller, Dan Focht, Geoff McAuliffe, and Alexander Greene.
I appreciate, as always, having access to such a generous group of people, who find the time to provide useful information. Thanks again!
Dwight Beebe E-mail: beebed-at-ere.umontreal.ca Institut de recherche en biologie vegetale Voice: 514-872-4563 Universite de Montreal FAX: 514-872-9406 4101, rue Sherbrooke est Montreal, Quebec H1X 2B2 Canada
So it takes more than "three strong people" to push them under? I'll bear that in mind!
We are still running an Edwards 12E6 coating unit coming up to 32 years old (can anybody beat that?), it goes like a Trojan (great for ploughing, farming joke!).
Regards - Keith Ryan Plymouth Marine Laboratory, UK
Perhaps I state the obvious, but beware of (powder) lubricated gloves. When I started SEM work (long, long ago), only powdered gloves were used for specimen prep. - what a mess! It seemed to always be "snowing" on samples sent to me from our failure analysis lab! Although somewhat difficult to locate at the time, I managed to convince all to go with powder-free. No more problem. Powder free are now easy to find.....
I know you have received alot of replies about this subject, but I can't resist adding my experiences. I compare having a water sprinkler system in an EM lab to gambling in Vegas; sometimes you win, sometimes you lose. We have a sprinkler system directly over our Cambridge 200 SEM, and fortunately we have not had to use it in 10 years - doesn't mean it won't happen tomorrow, though. We DID, however, have our ceiling cave in on our scope during a heavy rain. The soggy composite tiles were like chewed up spitwads all over the unit, and a steady river of water was running from the roof directly into the keyboard and console. Figuring that it was probably dead, I called the manufacturer for advise. We fixed the leak, then opened the SEM up and dried everything the best we could with blasts from a nitrogen tank. We allowed it to dry for 3 more days and fired it up. No problems other than some sticky keys in the keyboard - pretty amazing.
(BTW, this is not an advertisement for Cambridge/LEO. It's a real experience)
Regards,
-Bob ******************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com ******************************** ______________________________ Reply Separator _________________________________
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We recently moved our TEM and SEM from a building built in 1930 to one built in 1995. In other words, to a building with sprinklers. Now each microscope has a sprinkler directly overhead and I've been wondering if I should cover the scopes at night -- just in case. Do other people have this concern/problem? I haven't read anything about sprinklers malfunctioning and going off but it is a bit unnerving seeing a shower head directly over the scopes! I haven't seen covers in any of the catalogs so assume a tarp is the way to go....Or is it?
Thanks for any feedback....
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
This world wide E-mail provides important information for everyone involved in biomedical research. We hope no one feels bothered by this mail. Spamming is not indented.
Dear researcher,
Carl Zeiss is very pleased to announce it's new high performance confocal
Laser Scanning Microscope LSM 510,
designed for your best results in biomedical research.
(L)et's (S)ee (M)ore for applications, technical data and world wide expert contacts
at http://www.zeiss.de/mi/limi_e/p4/lsm510_entry.htm
This world wide E-mail provides important information for everyone involved in biomedical research. We hope no one feels bothered by this mail. Spamming is not indented.
Dear researcher,
Carl Zeiss is very pleased to announce it's new high performance confocal
Laser Scanning Microscope LSM 510,
designed for your best results in biomedical research.
(L)et's (S)ee (M)ore for applications, technical data and world wide expert contacts
at http://www.zeiss.de/mi/limi_e/p4/lsm510_entry.htm
As an anthropological electron probe microanalytical (EPMA) project we are analyzing ancient and diagenetic bone for strontium, barium and other trace elements. We are seeing an unexpected (so what else is new) correlation of Sr with phosphate and the i nverse with apatite replaced by carbonate. We are trying to eliminate analytical errors, and would like to throw this problem at the EPMA community for feedback ... my standard apatite with no Sr measures at detection, and we have double-checked for backg round interferences, but only for elements suspected. We are suspicious of not analyzing for a element which might be causing this problem ... TIA
cheerios, shaf
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
Does anyone know of a good preferential etch for P+ silicon? I've been using a 20:1 HNO3:HF stain. This stain seems to delineate the N+ areas very well, but the P+ areas are not easily seen.
I would like to decorate the P+ with as little attack to the N+ areas as possible.
Any help is greatly appreciated.
Regards, Bob Heiman ------------------------------------- E-mail: heiman-at-gmtme.com Voice: 610-666-7950 x2855, FA Lab x2533
Message-Id: {199703031915.OAA14276-at-post-ofc04.srv.cis.pitt.edu} To: microscopy {microscopy-at-Sparc5.Microscopy.Com}
-- [ From: Simon Watkins * EMC.Ver #2.5.02 ] --
As another tale of woe to follow up on Bob Citrons comment on soggy ceiling tiles forming spit wads all over his SEM.
A few years back while I was working in Boston we tried to drown a 100CX by backing up several gallons of water down an air handling chute directly on top of the column (don't ask why or how, but it was one of the side effects of being in the basement). Being on the ball, the operator recognised the problem due to the high moisture content in the air and the splishy splashy sound of water falling from the ceiling. He hit the magic red button to shut the instrument down, and ran for help shouting as he ran "the microscopes drowning, the micrscopes drowning".
As was the case with Bob we called the manufacturer, in this case the ever helpful folks at JEOL, who agreed this was something of a problem but had no immediate solution beyond "errrr, huh, interesting", so we dried off what we could and left the thing off for a week. We turned it on and away it went, as if nothing had happened. The only long term effect was some rusting of the Allen bolts used to hold the column together, you would think that they would be painted or stainless with all these "environmental" problems of late!
my 2 cents
Simon
-- ------------------------------------------------- Simon C. Watkins Ph.D Associate Professor Director Center for Biologic Imaging University of Pittsburgh Pittsburgh PA 15261 tel 412-648-3051 fax 412-648-2004 -----------------------------------------------
The book, Handbook of Metal Etchants, has about a hundred pages of etchant recipes for Si wafers, crystals. Good discussions on practices/ applications as well. Includes references.
Handbook of Metal Etchants Walker, Perrin & Tarn, William H. 1991 CRC Press, Boca Raton, FL 33531 ISBN 0-8493-3623-6
Happy mixing! ------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
Thanks to all who answered my recent question about covering EM's against sprinkler damage! A lot of good points were made, including:
1. The threat is real (some real horror stories out there!) 2. It's not just sprinklers that threaten, it's floods from burst plumbing, storms etc... 3. Tarps are inconvenient and possibly hazardous, 4. Curved plexiglass or fiberglass structures attached to the ceilings over the microscopes were widely recommended (like salad bars) 5. Many suggested dismantling or plugging the sprinkler heads while 6. others were concerned about fire safety 7. A surprising number of drenched scopes fully recovered 8 which you should keep in mind if you actually push your scope under the downpour in hopes of obtaining a new one. 9. but if you try anyway, it takes more than three people.
Thanks again!
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
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Subject: Time:1:27 PM OFFICE MEMO Why am I not surprised? Date:3/3/97
Last week I received a message from Microscopy about "Little Jessica Mydeck". A check at http://www.cancer.org/acs.html revealed:
Fraudulent Chain Letter (This statement may be copied or reprinted by online users)
The American Cancer Society is greatly disturbed by reports of a fraudulent chain letter circulating on the internet which lists the American Cancer Society as a "corporate sponsor" but which has in no way been endorsed by the American Cancer Society.
This letter appears to have started on America Online but has now spread well beyond the online service. There are several variations of this letter in circulation. The text of the original message reads as follows:
LITTLE JESSICA MYDEK IS SEVEN YEARS OLD AND IS SUFFERING FROM AN ACUTE AND VERY RARE CASE OF CEREBRAL CARCINOMA. THIS CONDITION CAUSES SEVERE MALIGNANT BRAIN TUMORS AND IS A TERMINAL ILLNESS. THE DOCTORS HAVE GIVEN HER SIX MONTHS TO LIVE.
AS PART OF HER DYING WISH, SHE WANTED TO START A CHAIN LETTER TO INFORM PEOPLE OF THIS CONDITION AND TO SEND PEOPLE THE MESSAGE TO LIVE LIFE TO THE FULLEST AND ENJOY EVERY MOMENT, A CHANCE THAT SHE WILL NEVER HAVE. FURTHERMORE, THE AMERICAN CANCER SOCIETY AND SEVERAL CORPORATE SPONSORS HAVE AGREED TO DONATE THREE CENTS TOWARD CONTINUING CANCER RESEARCH FOR EVERY NEW PERSON THAT GETS FORWARDED THIS MESSAGE. PLEASE GIVE JESSICA AND ALL CANCER VICTIMS A CHANCE.
IF THERE ARE ANY QUESTIONS, SEND THEM TO THE AMERICAN CANCER SOCIETY AT ACS-at-AOL.COM
As far as the American Cancer Society can determine, the story of Jessica Mydek is completely unsubstantiated. No fundraising efforts are being made by the American Cancer Society in her name or by the use of chain letters. Furthermore, the email address ACS-at-AOL.COM is inactive. Any messages to the American Cancer Society should be instead sent through the American Cancer Society website at http://www.cancer.org.
I would like to get an explanation for the following observation of quantitative analysis by EDS.
I am looking at a Si wafer with 100nm of SiO2 layer on top. from a SEM. Electron beam (~8 kev) hit the surface along the surface normal (0 deg tilt). As I increase the sample tilt (upto 45 deg or so) EDS quantitative analysis shows a decrease of O2%. How this is possible? I expected to see O2% goes up with the tilt as electron will see more of the surface than bulk.
Could someone explain why?
Thanks
Fran.
--------------------------------------------------------- Get Your *Web-Based* Free Email at http://www.hotmail.com ---------------------------------------------------------
Many thanks for the many recent replies to my earlier queries on Disposable gloves and multi user security. We now have a few more ideas on where to go regarding security in our Unit. The gloves issue is very complex too, mainly due the wide variety of chemicals and conditions they are used for in the lab!
Thanks again,
Rich.
----------------------------------------------------------------------- Richard Lander, NZCS South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
fransisco black wrote: } Dear Microscopists: } I would like to get an explanation for the following observation } of quantitative analysis by EDS. } } I am looking at a Si wafer with 100nm of SiO2 layer on top. } from a SEM. Electron beam (~8 kev) hit the surface along the } surface normal (0 deg tilt). As I increase the sample tilt } (upto 45 deg or so) EDS quantitative analysis shows a decrease } of O2%. How this is possible? I expected to see O2% goes up } with the tilt as electron will see more of the surface than } bulk. } Could someone explain why? } Thanks } Fran.
Fran, My initial guess would be the change in the sample-detector geometry. Although one would expect the O2 counts to go up, the detector may no longer be in as good a position (relative to the sample) to pick them up. You might simply adjust the insertion of the EDS detector to compensate.
One could use an interpolative process: adjust the tilt a bit, then adjust the detector insertion to maximize counts. Then adjust the tilt a little more, and readjust the detector insertion.
You might get to an optimum geometry faster if you do a rough calculation of the angle of incidence, reflection and best detector insertion to intersect.
Regards, John Best -- ELMDAS Co. http://www.vicon.net/~jbest
} I am looking at a Si wafer with 100nm of SiO2 layer on top. } from a SEM. Electron beam (~8 kev) hit the surface along the } surface normal (0 deg tilt). As I increase the sample tilt } (upto 45 deg or so) EDS quantitative analysis shows a decrease } of O2%. How this is possible? I expected to see O2% goes up } with the tilt as electron will see more of the surface than } bulk. } Dear Fran, What is the geometry (take-off angle, direction of tilt, etc.)? It may be that some O x-rays are taking a longer path and are absorbed, or the Si x-rays can take a shorter path and be absorbed less. Have you used David Joy's Monte Carlo program to calculate the volume in which the x-rays are generated? Yours, Bill Tivol
All else being the same, I (as it sounds you would) would expect an increase in the relative intensity of the oxygen (raw counts). Do you see this on the graphical result? If so, it is most likely that the ZAF/PhiRhoZ/etc. quant.
algorithm is the source of the problem. Woody ______________________________ Reply Separator _________________________________
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Dear Microscopists:
I would like to get an explanation for the following observation of quantitative analysis by EDS.
I am looking at a Si wafer with 100nm of SiO2 layer on top. from a SEM. Electron beam (~8 kev) hit the surface along the surface normal (0 deg tilt). As I increase the sample tilt (upto 45 deg or so) EDS quantitative analysis shows a decrease of O2%. How this is possible? I expected to see O2% goes up with the tilt as electron will see more of the surface than bulk.
Could someone explain why?
Thanks
Fran.
--------------------------------------------------------- Get Your *Web-Based* Free Email at http://www.hotmail.com ---------------------------------------------------------
I teach electron microscopy (both TEM and SEM) as a one semester course. The first couple of times I had 3 to 8 students but last fall I had 14. Students used photographic film and paper out of a common supply. It seemed that every time I turned around, we were out of paper and film or some unidentified person had exposed a portion of our common supply to light. In order to better monitor usage and I am considering giving each student his/her own allotment of paper and film at the beginning of the semester. I require them to turn in 10 TEMs and 10 SEMs at the end of the semester. What do those of you who teach EM (and old fashioned darkroom techniques) require for your students to turn in and how do you budget and/or distribute supplies?
Bob
Robert R. Wise Plant Physiologist and Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
} Dear Microscopists: } } I would like to get an explanation for the following observation } of quantitative analysis by EDS. } } I am looking at a Si wafer with 100nm of SiO2 layer on top. } from a SEM. Electron beam (~8 kev) hit the surface along the } surface normal (0 deg tilt). As I increase the sample tilt } (upto 45 deg or so) EDS quantitative analysis shows a decrease } of O2%. How this is possible? I expected to see O2% goes up } with the tilt as electron will see more of the surface than } bulk. } } Could someone explain why? } } Thanks } } Fran. }
The problem observed here is most likely related to simple rules of geometry. If the EDS detector is not perpendicular to the tilt axis you will find that the path through the sample that the xrays must use to get to the EDS detector becomes longer as the tilt axis increases. (Lower observed take off angle). This results in more absorption. The lower energy peaks will be abosrbed at a greater rate than the higher energy lines. If you are not taking the azimuth angle into your take off angle correction you should.
If you want to limit your xray excitation volume to give you a better idea of the film try also using a lower accelerating voltage. Normally an excitation energy of 2.5 times your xray energy should work.
BTW: I have a DTSA binary -} EMMFF ascii file converter written in Python if anyone needs one. It actually writes a somewhat loose version -- the spec was ambiguous in several places and I loosened some restrictions intentionally for my own purposes. It's written in Python, and it's easily modifiable to your own customizations. ( Specifically, I use longer user defined keyword fields to save the original DTSA header information using the names in the DTSA file interface definition. The DTSA EMMFF plugin seems to read these files back in properly -- the standard tags that is. It ignores the original saved DTSA information. )
I wrote this code before there was a plug-in for DTSA, however I still require it to do batch conversions of folders full of files.
The current version is specifically for the Mac. The initial version was originally run on unix, and most of it should be portable. The main mac specific feature is that it supports drop launching of a whole folder of DTSA files. When I have the final, portable version ( hopefully with a stand-alone Mac version that doesn't require Python installed. ) it will be posted on the Web. If anyone needs it right away, I can email you a copy of the sources.
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |--- By doing just a little every day, you can gradually let the task completely overwhelm you.
I am a student of EM at Madison Area Tech. We had to buy our own paper. I am now a lot better at making prints !
Leah
---------- } From: wise-at-vaxa.cis.uwosh.edu } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Supplies and teaching } Date: Tuesday, March 04, 1997 9:13 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } To all, } } I teach electron microscopy (both TEM and SEM) as a one semester } course. The first couple of times I had 3 to 8 students but last fall I } had 14. Students used photographic film and paper out of a common supply. } It seemed that every time I turned around, we were out of paper and film or } some unidentified person had exposed a portion of our common supply to } light. In order to better monitor usage and I am considering giving each } student his/her own allotment of paper and film at the beginning of the } semester. I require them to turn in 10 TEMs and 10 SEMs at the end of the } semester. } What do those of you who teach EM (and old fashioned darkroom } techniques) require for your students to turn in and how do you budget } and/or distribute supplies? } } Bob } } } Robert R. Wise } Plant Physiologist and Director, UWO Electron Microscope Facility } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-uwosh.edu } }
Greetings, I have a user of our EM Unit who wishes to use Fluoronanogold (Nanoprobes) with Silver enhancement for pre-embedding immuno EM. We will be using frozen 8 micron sections which will then be PLP fixed (parform + Lysine + periodate) prior to labelling. After labelling, we want to post-fix them in OsO4 (as membranes are important), dehydrate and embed in normal epoxy resin. I have heard that the Flouronanogold is not stable at 60oC, the normal curing temperature of the resin.
Has anybody had experience with pre-embedding with Fluoronanogold? Will Fluoronanogold survive the epoxy resin curing process at 60oC? If it wont survive, which resin and curing procedure would be the choice to use in this case? (Quetol 651, Lowicryl?)
I am interested to replace our SEM by ESEM or low vacuum SEM, but I would like to have more information about EDX microanalysis with these SEM (what are the quantitative and qualitative differences when I change the pressure? what's happen with cryo system .....?). Thanks in advance for your help (comments, references ...)
Best regards to all Didier
-------------------------------------------- Dr. Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
First, my apologies to the Microscopy listserver subscribers if this posting isn't appropriate. We need a first rate science teacher with experience, and my colleagues here at the microscopy listserver sprung to mind immediately.
Secondly, If your not interested in accepting a position as a high school science teacher, and know of a listserver that may produce some interested candidates, please forward this request. Thank You.
And finally: The Juniata Valley School District is seeking candidates for the position of Science Teacher beginning in the fall of 1997. Responsibilities are: fundamentals of science and physical sciences for students in grades 7-12.
We're looking for candidates who bring a number of years of experience from industry or a university research environment to the classroom. Pennsylvania teaching certification is required, or must be readily obtainable. The ability to motivate and educate "typical" high school students is a must.
With a BS the starting salarary is $28,041, an MS starts at $28,712. Starting salaries are dependant on teaching experience within the Pennsylvania public school system.
The Juniata Valley School District has approximately 500 students in grades 7-12 and is situated in a beautiful rural area approximately 25 miles southwest of the main campus of The Pennsylvania State University. JVSD is an equal opportunity employer.
Please forward your resume to our superintendant, Dr. David Leckvarcik at The Juniata Valley School District, P.O. Box 318, Alexandria, PA 16611. If you would like me to follow up on your resume, please forward it to me at: jbest-at-vicon.net.
I am wondering what solutions people have found to the stage temperature/gas/humidity control problem for tissue culture microscopy of live cells. I have tried an open chamber approach (20/20 Technologies) and evaporation onto the condenser or lid was a major problem as was even heating. Are there any commercial stage temperature controllers that rely on hooding the microscope or stage that work well? Has anyone constructed something simple that can be reproduced easily? What are the down sides of this approach? Has anyone used the new Bioptechs system and does it actually do the job and solve any or all of these problems? Thanks- Dave
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu
I am looking for a PC based software (preferably on Windows) that can perform profilometry type measurements from an SEM pair of images (grabbed into 'TIF' files). Measurements should include: - Line profiles - Height measurements - Contour maps.
Any help is appreciated.
Regards,
Dr. Rafik Allem Pulp and Paper Research Institute of Canada. 570, St. John's Boulevard, Pointe-Claire, Qc, H9R 3J9, Canada. tel: (514)630-4101 ext. 2661, fax:(514)630-4134 e-mail: allem-at-paprican.ca
Department of Mining and Metallurgical Engineering Professional Development Seminars
"MATERIALS CHARACTERIZATION FOR THE METAL, MINERAL AND MICROELECTRONICS INDUSTRY" May 12-15, 1997
Seminar Leader: Neil Rowlands
This course is intended to benefit users who are concerned with the analysis of fine structures, fine particles and may also have the need to image surfaces down to the atomic level. These techniques will be of special interest to those involved in the microelectronics, polymer science, and ceramics industries, as well as those involved in metallurgical analysis.
The course will be of a practical nature and will be of interest to operators and managers of analytical laboratories R&D engineers and applied scientists. Examples of the applications of each technique will be presented and on the last day of the course there will be an optional visit to the Materials Technology Laboratory, CANMET in Ottawa to see many of these techniques in operation. Additional facilities (e.g. TEM, AFM) are available on the McGill campus.
I already replied privately before I got down our in-box to the MSA posting. For anyone interested in this, here are our ruminations...
Generally, we recommend low-temperature polymerization resins such as Lowicryl, LR White or any similar medium for embedding Nanogold=81 or =46luoroNanogold-labeled specimens. This is because some early experiments with Nanogold=81-labeled specimens suggested that the higher-temperature embedding (60=B0C) might cause the gold particles to be displaced from their binding sites. However, subsequent experiments with Nanogold=81 in solution showed that it could be heated even up to 100=B0C for 1 h with minimal decrease in optical density; generally, avoiding low pH values (below 7) or high ionic strengths (0.2 M NaCl or higher) helps ensure its stability. Because it is smaller than most colloidal gold and has no tendency to stick electrostatically to proteins or cell components, it may be more free to move: fixing with glutaraldehyde helps counteract this, and with Nanogold=81 we also recommend silver enhancement before embedding if it is practical.
There is a section on this in Hainfeld and Furuya's chapter on the silver enhancement of Nanogold=81 and undecagold in M. A. Hayat's recent book, "Immunogold-Silver Staining: Principles, Methods and Applications" (CRC Press, Boca Raton, 1995; pp. 71-96. Check pages 92-92 for the effects of heating Nanogold=81. From this section, heating at 60=B0C for 250 minutes resulted in a reduction of the optical density to 80% of its initial value - suggesting that Nanogold=81 can survive most 60=B0C embedding procedures.
The gold particle in FluoroNanogold is Nanogold, and exactly the same applies to this probe. However, silver enhancement quickly removes the fluorescence, so only do the pre-embedding silver enhancement if you have completed the fluorescence microscopy.
We keep a list of answers and suggestions to frequently-asked questions in the Technical Help section of our web site:
http://www.nanoprobes.com/Tech.html
All the suggestions made there about Nanogold=81 use and stability also appl= y to Fluoronanogold, since these probes use the same gold particle.
Hope this is helpful,
Rick Powell
****************************************************************** * NANOPROBES, Incorporated | Tel: (516) 444-8815 * * 25 East Loop Road, Suite 124 | Fax: (516) 444-8816 * * Stony Brook, NY 11790-3350, USA | nano-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
In response to Allan Mitchell's inquiry. If you are speaking of whether the Fluorescent signal of Fluoro-Nanogoold survives the answer is no, but not because of the 60 degree heat. The fluorescence is quenched or covered up by the silver enhancement process. If you want to see the label prior to silver enhancement that is no problem. I often do this prior to running the silver enhancement to see that the reagent has gotten in to the sample and gives the expected signal. Your other choice of resins are lowicryl which you can polymerize at -35 degrees under UV. The FLUOR tag survives this just fine. In regard to osmication, this has to be done with 0.1% osmium at 4 degrees or on ice for about 30 - 45 minutes. Standard osmication procedures will reduce the silver shell that forms around the nanogold particle. This appears as a much less electron dense cloud or ghost. I have used these reagents with spurr, epox 812, lowicryl and LR White without any problem.
Joe Goodhouse Confocal / E.M. Lab Molecular Biology Princeton University
} From: wise-at-vaxa.cis.uwosh.edu } Date: Tue, 04 Mar 1997 09:13:27 +0000 } Subject: Supplies and teaching } To: Microscopy-at-Sparc5.Microscopy.Com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } To all, } } I teach electron microscopy (both TEM and SEM) as a one semester } course. The first couple of times I had 3 to 8 students but last fall I } had 14. Students used photographic film and paper out of a common supply. } It seemed that every time I turned around, we were out of paper and film or } some unidentified person had exposed a portion of our common supply to } light. In order to better monitor usage and I am considering giving each } student his/her own allotment of paper and film at the beginning of the } semester. I require them to turn in 10 TEMs and 10 SEMs at the end of the } semester. } What do those of you who teach EM (and old fashioned darkroom } techniques) require for your students to turn in and how do you budget } and/or distribute supplies? } } Bob } } } Robert R. Wise } Plant Physiologist and Director, UWO Electron Microscope Facility } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-uwosh.edu } } } Bob,
We here at BYU teach a course in SEM and in TEM. We require our students to submit two types of assignments to complete the course. First is a portfolio which may or may not use traditional darkroom techniques (digitized, laser printed images are an option from our SEMs). We also have the students turn in a poster representing a project they have worked on all semester. The poster must have at least 5 high quality photographs which the student has produced, as well as any text and graphics the student wants to display. The student purchases all of the photographic materials they use. We do keep a stock supply in the lab for the students to purchase from. Students are responsible for all of their own photographic supplies so if thier paper becomes light struck, that is their problem. This has worked quite successfully for the past several years.
I hope this helps
Michael D. Standing e-mail: MDStandi-at-bioag.byu.edu Phone: (801)378-4011
I have noted several users of this listserver refer to the Ilford RC2150 print processor. We have used one for several years without trouble but Ilford has apparently switched to a new "environment friendly" cleaning solution that is unable to remove the deposits from the rollers. All of our prints have terrible streaks of silver residue. Has anybody else had this problem - more importantly, has any been else solved it? TIA
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Way back when, the TEM class I took, it was required for me to purschase my own film, paper, tweezers, embedding mold and a text book. All this was setup by the proffessor and sold thru the bookstore on campus. Yes it was/is expensive but it taught me to be conservative with these items. Most of the prints I made for the class were 4x5. The class was ten weeks long and I did not run out of supplies.
Hopes this helps,
Ed Calomeni Dept. Pathology Medical College of Ohio Toledo, Ohio emlab-at-opus.mco.edu
I am trying to use stains discribed in Histochemistry: Theoretical and Applied Volume 2 by AGE Pearse to locate metallic ions with in tissue. Any suggestions for other methods will be appreciated. The following are the stains, the metal I'm trying to detect, and the problem I am having. I trying to stain cells not sections.
Napthochrome Green B Beryllium (1)unable to find acridine red Aldridge says is was discontinued 12 years ago
Rubeanic Acid Nickel (1) the stain is very light and is not staining cells
Chrome Azurol S Chromium (1)library is unable to get referenced paper and therefore I am unable to find protocol
If anyone has any information that might help please responed.
Regina Messer Graduate Student Biomedical Engineering UAB
For staining metals look in "Histopathologic Technique and Practical Histochemistry" by Ralph Lillie and Harold Fullmer; McGraw Hill, 1976. Out of print for years but probable available in your library or via interlibrary loan. Also "Selected Histopathological Methods" or something like that by Thompson is a massive tome with just about everything in it. Call or e-mail if necessary.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
We have observed that spatial x-ray resolution suffers, even at 40 Pa with a 25 mm working distance. We can get decent x-ray maps, but quantitative analyses can easily pick up tenths of a percent of an element from a nearby phase. We have yet to quantify the effects for ourselves. I recall there were a number of papers at the Minneapolis meeting on this effect. One has to be careful.
At 10:30 AM 3/5/97 -0500, you wrote:
} Dear all, } } I am interested to replace our SEM by ESEM or low vacuum SEM, but I } would like to have more information about EDX microanalysis with these SEM } (what are the quantitative and qualitative differences when I change the } pressure? what's happen with cryo system .....?). } Thanks in advance for your help (comments, references ...) } } Best regards to all } Didier } } -------------------------------------------- } Dr. Didier Le Thiec ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
Hello: I am looking for a used SEM with low Voltage capability, about 1 KV. Let me know if you know of anybody who would like to sell. Please E-mail or call me at 909 694-1839. Thanx, Peter Jordan
Didier Le Thiec wrote: } I am interested to replace our SEM by ESEM or low vacuum SEM, but I } would like to have more information about EDX microanalysis with } these SEM (what are the quantitative and qualitative differences when } I change the pressure? what's happen with cryo system.....?)
Dear Didier,
The presence of gas in the specimen chamber influences the spatial resolution of EDX microanalysis in ESEM and LVSEM. The primary electrons will scatter on the gas and give rise to a skirt of scattered electrons that will excite X-rays at places pretty far away from the point you want to analyze. The scattered intensity is approximately given by Is/Io = 1 - exp(-psL/kT) where p is the pressure, s the total scattering cross section for electron scattering on the gas used, L the distance between the last pressure limiting aperture and the sample, k the Boltzmann constant and T the absolute temperature. Examples of skirt shapes are given in:
D. A. Moncrieff et al., J. Phys. D: Appl. Phys. vol. 12 (1979) 481-88. D. C. Joy, Microscopy and Microanalysis ' 96, Proc. Annual Meeting MSA, Minneapolis, Minnesota, 11 - 15 August 1996
It is to a large degree possible to correct for the beam skirt effects:
1) You can extrapolate from spectral measurements made at several different pressures to the result that would have been found without scattering provided that the measurements are made in the single scattering regime (e.g. pL { approx. 1.6 Pa.m for measurements in water vapour). 2) If there is plural scattering, you can take two spectra, one with a fine needle (of the kind used for field ion microscopy or scanning tunneling microscopy) inserted over the point of interest, and the other with the needle slightly retracted. Subtraction of the first from the second spectrum will approximately give the spectrum from the point of interest.
Neither method will give you as exact an analysis as you will get under high vacuum, but you can get rid of most of the skirt effects. The pressure variation method in particular yields pretty good results if carefully performed. The methods are described in:
J. B. Bilde-Soerensen and C. C. Appel, Proc. 48th Annual Meeting of the Scandinavian Society for Electron Microscopy, Aarhus, 2 - 5 June 1996, pp. 4 - 5. J. B. Bilde-Soerensen and C. C. Appel, Proc. 11th European Congress on Microscopy EUREM' 96, Dublin, 26-30 August 1996. Session T6.
ESEMs and LVSEMs can also be operated as high vacuum SEMs - so you still have the possibility of going to high vacuum for microanalysis.
Best wishes, Jorgen. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- J. B. Bilde-Soerensen Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
We have published a couple of papers on ESEM EDX. Might be worth a look. :) I would like to add that if you wish to look at hydrated samples then the chamber pressures required are such that scattering of the primary beam results in a probe at the sample of at least 1mm diameter!! Also correction methods based on taking spectra at different pressures are fraught with difficulty if you need to avoid drying out your sample. Also be aware that with low Z detectors a contribution from the chamber gas will be encountered.
Chris
} I am interested to replace our SEM by ESEM or low vacuum SEM, but I } would like to have more information about EDX microanalysis with these SEM } (what are the quantitative and qualitative differences when I change the } pressure? what's happen with cryo system .....?). } Thanks in advance for your help (comments, references ...) } } Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171
At 02:43 PM 3/5/97, Hildagonda van der Merwe wrote: } Can anyone please tell me all the uses of the Immuno-Bed Kit?
Immunobed is an embedding medium for immunohistochemistry which allows the rapid penetration of large immunoglobulins through the section for demonstration of antigenic sites.
For more detailed information, please request our A2050 data sheet.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
I wish to thank those people (Scott Whittaker, Gary Chinga, Christian Mellen, Melvyn Dickson, John Mardinly, Hans Tietz) who replied me, through there were a few. (Sprinkling water seems to be much more interesting to talk about.:) Short summary is: The main software for image analysis is NIH (FTP from zippy.nimh.nih.gov), through analySIS also exists and Photoshop, Paintshop, Coreldraw and Wincatpro can be used to manipulate images.
CD-Rs mainly used to store, transport and distribute image files.
1200dpi LP (mainly Lexmark) used for draft and dye-sub printers for quality prints. So one will need both...
Useful references obtained: http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks " site supported by Scott Whittaker - many discussions from this Listserver and Confocal Listserver archived there. Extremely useful!
http://www.soft-imaging-web.de home of Soft-Imaging Software information about analiSIS software package, etc.
Stewart, M.G. and Davies, H.A. (1995) Digital image processing (DIP) in the TEM: is it viable in biological morphometry?. Eur Microsc Anal (1995) 35, pp 21-23.
Special opinion: "Andrej! Do not change!..."
Full text of replies can be obtained on request. A. Chuvilin
I saw your concern about the EDX at higher pressures. As is suggested by others there are inaccuracies in the quantitative estimation. This is because the incident beam coming through the gaseous region spreads before it hits the specimen surface. The electrons outside the original gaussian profile of the beam also create x-rays. You are probably aware of David Joy's work which was presented at MSA meeting in the US. We are enormously interested in this problem and one of my students is doing Monte carlo calculations on this very problem. You also inquired about cryoSEM. Normally, you should not have any problem about the beam scattering as the equilibrium vapour pressure is below the chamber pressure. So you can assume that all of your x-rays are coming from the original focussed beam spot. This of course assumes that you are not using a very intense beam which evaporates water or other constituents in your specimen and upsetting the inherent composition of the specimen. Incidentally,at Bristol we convert conventional SEMs in to High pressure SEMs exceeding the performance of the commercially avilable instruments. If you need any help on this please do not hesitate to get in touch with me
Yours sincerely
Jitu Shah
Dr.Jitu Shah H.H. Wills Physics Laboratory, University of Bristol, Royal Fort, Tyndall Avenue, Bristol BS8 1TL. UK email: jss-at-siva.bristol.ac.uk Tel: 44 117 9288719 Fax: 44 117 9255624
When I perform an EDX analysis on uncoated fused silica, 10 20 or 30 KeV, and the sample begins to charge, often I get a small aluminum peak, and it doesn't go away as I move around the sample, it comes back. The amount of aluminum in the quartz is typically less than 20 ppm, so I don't think that the Al peak is coming from my sample. Could it be that the Al peak is coming from my sample stage or walls of the sample chamber, due to any effect caused by the charge that has developed on the uncoated quartz? Also, for EDX work, what are the negative effects that sample charging has on an analysis? My work is done on an Amray 1600 with a Noran detector, one that has two windows, Be and the ultra thin window.
Recently I got a problem in LVSEM-EDS system. The SEM is JSM-5400LV with Kevex EDS. We use a thin B-window with expect to determine elements down to Z=5. The SEM does not have pre-vacuum chamber which means whenever we change sample, the chamber and whole column are exposured to atmosphere. The window has been used for 10 months with good performance. Therefore, recently once we change sample as usual, the window got brocken. We have no idea what is the reason.
My question is: How thick window should I use to prevent the window from brocken, and what is the common technical hints to prevent the window from brocken? Does the frequently presure change on window surface (when change sample) affect the life time of window?
Thank you very much
Zhiyu Wang Electron Microscope Facility Western Kentucky University Bowling Green KY 42103 USA wangz-at-pulsar.cs.wku.edu
I am trying to do trace Carbon analysis in Nb material. The Be window of my EDX can be opened in order to detect C, N, O, but the quantification seems impossible. Anybody has similar experience?
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Hello Everyone,
I have been tasked with bringing another technician into our SEM lab and would like to get info on the courses which are available for a person who is familiar with light microscopy but knows almost nothing about a SEM.
The desired course would be 5 days or less and be applicable to the level of knowledge described above. We deal with strictly materials analysis, so biological based courses are not applicable. The desired outcome would be someone who has a rudimentary knowledge of SEM theory and capable of taking basic SEM pictures and acquiring EDS spectra on non-challenging samples. We will be doing some training of the new tech, but due to heavy workloads, we can't devote the time to the training that we would normally do.
Geographically, I would like courses in the Southeastern USA, but will take info on courses anywhere in the US.
John Giles Senior Materials Engineer Honeywell Space Systems Clearwater, FL
The EM facility at EPA-Cincinnati is hopefully going to replace a 20 year old critical point dryer in the near future. If anyone has any recommendations, or horror stories, let us know before we spend your tax dollars. You can reply to us directly at clark.patrick-at-epamail.epa.gov Thanks
This happened to me once, and it turned out to be stray x-rays from the EDX detector collimator sleeve. If your detector has one of these, slide it off and paint the inside surfaces with carbon dag and try it again; it may solve your problem.
Regards,
-Bob ********************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com **********************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When I perform an EDX analysis on uncoated fused silica, 10 20 or 30 KeV, and the sample begins to charge, often I get a small aluminum peak, and it doesn't go away as I move around the sample, it comes back. The amount of aluminum in the quartz is typically less than 20 ppm, so I don't think that the Al peak is coming from my sample. Could it be that the Al peak is coming from my sample stage or walls of the sample chamber, due to any effect caused by the charge that has developed on the uncoated quartz? Also, for EDX work, what are the negative effects that sample charging has on an analysis? My work is done on an Amray 1600 with a Noran detector, one that has two windows, Be and the ultra thin window.
Would you be so kind as to cite your literature references, so we can read them. They sound useful for a number of us doing EDX in a variable pressure SEM. Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Looking for image analysis software/solutions for the following:
1} sperm analysis,cell motility etc. 2} textile analysis,rayon fibre analysis,broken fibre etc.
Could anyone point out some good manufacturers/sources with address/email/fax wwith name of source.?////// I would be grateful if someone could help me in this context.
Thanks,
Best regards,
Anish
************************************************************************* For further details please contact: Soneja A.K. Director METZER BIOMEDICAL & ELECTRONICS PVT.LTD. 327 Wadala Udyog Bhavan,Wadala,MUMBAI(BOMBAY )400 031.INDIA Tel:91 22 4145057/4165650 Fax 91 22 4168757
There are a couple of reasons why windows can have a limited lifetime when used in LV-SEM & ESEM.
Firstly, when the chamber is pumped to and from atmosphere any loose particles in the chamber are disturbed, and can damage the window by hitting it. This can be reduced by fitting some kind of loose cover (we use a plastic bag!) over the x-ray probe whenever using the microscope for non x-ray work.
Secondly, certain windows when used in lo-vac are susceptible to ice formation on the surface and actually in the window.
Regarding probe-beam spreading, we, like Dr. Shah, are carrying out monte-carlo simulations, in conjunction with x-ray measurements and will hopefully be presenting some results at this years MSA meeting
Ian Bache Cavendish Laboratory Cambridge University Madingley Road Cambridge ENGLAND (+44)-1223-337229
} Would you be so kind as to cite your literature references, so we can read } them. } They sound useful for a number of us doing EDX in a variable pressure SEM. } Thanks. } Sorry I should have included them on the original post!
Sigee, D.C. and Gilpin, C.J. X-ray microanalysis with the environmental scanning electron micrasocpe: Interpretation of data obtained under different atmospheric conditions. Scanning microscopy supplemnet 8 219-229 1994
X-ray microanalysis of wet biological specimens in the environmental scanning electron microscope: 1 Reduction of specimen distance under different atmospheric conditions. Journal of Microscopy 179 (1) 22-28 1995
More work in progress as they say!
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171
} Would you be so kind as to cite your literature references, so we can read } them. } They sound useful for a number of us doing EDX in a variable pressure SEM. } Thanks. } Sorry I should have included them on the original post!
Sigee, D.C. and Gilpin, C.J. X-ray microanalysis with the environmental scanning electron micrasocpe: Interpretation of data obtained under different atmospheric conditions. Scanning microscopy supplemnet 8 219-229 1994
X-ray microanalysis of wet biological specimens in the environmental scanning electron microscope: 1 Reduction of specimen distance under different atmospheric conditions. Journal of Microscopy 179 (1) 22-28 1995
More work in progress as they say!
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171
John Giles wrote: ===============================================
I have been tasked with bringing another technician into our SEM lab and would like to get info on the courses which are available for a person who is familiar with light microscopy but knows almost nothing about a SEM.
The desired course would be 5 days or less and be applicable to the level of knowledge described above. We deal with strictly materials analysis, so biological based courses are not applicable. The desired outcome would be someone who has a rudimentary knowledge of SEM theory and capable of taking basic SEM pictures and acquiring EDS spectra on non-challenging samples. We will be doing some training of the new tech, but due to heavy workloads, we can't devote the time to the training that we would normally do. ================================================= We would give extremely high marks to the "Lehigh" courses which are given at Lehigh University, Bethlehem, PA every year. You can get information about their program from their website at the following:
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We anticipated window problems from a detector left in the wet environment 24/7 and cycled for every sample change. We initially had a hard time finding an EDS vendor to build us a detector that was fully retractable behind a gate valve outside the 2020 chamber so that the detector was protected not only from projectiles and cycles but from harsh environments sometimes used in environmental scopes. We did eventually get such a detector and had window problems initially but that seems to have been solved with a slightly thicker window. My advice is that everyone should let their EDS reps know that this type of detector is desirable and maybe one day they will be widely available.
I also wonder if the commercially available window materials are water tight. As we all know one of the EDS vendors uses a heater to remove ice from its crystal. Helium leak tests are often quoted when asked about water permeability but I would like to see water leak test data. If water is geting into and through windows it could be responsible for lost windows in wet systems.
I am looking forward to these monte carlo papers and lively discussions at the M&M meeting in Clevland.
Scott Wight
} There are a couple of reasons why windows can have a limited lifetime when } used in LV-SEM & ESEM. } } Firstly, when the chamber is pumped to and from atmosphere any loose } particles in the chamber are disturbed, and can damage the window by } hitting it. This can be reduced by fitting some kind of loose cover (we } use a plastic bag!) over the x-ray probe whenever using the microscope for } non x-ray work. } } Secondly, certain windows when used in lo-vac are susceptible to ice } formation on the surface and actually in the window. } } Regarding probe-beam spreading, we, like Dr. Shah, are carrying out } monte-carlo simulations, in conjunction with x-ray measurements and will } hopefully be presenting some results at this years MSA meeting } } } } Ian Bache } Cavendish Laboratory } Cambridge University } Madingley Road } Cambridge } ENGLAND } (+44)-1223-337229
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
Try Visilog from Noesis Vision Inc, we have a turn key system available for sperm analysis and or fibre analysis. You can reach us at http://www.noesisvision.com
At 12:51 PM 3/7/97 +0530, SONEJA A K wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------------------------------------------------------------- --------------------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada
Visit our new web site at http://www.noesisvision.com ---------------------------------------------------------------------------- ---------------------
The Royal Microscopical Society Computers in Microscopy ,September 1997, Cambridge CALL FOR PAPERS
A Colloquium on various aspects of the application of computers to microscopes and microscopical techniques is being planned by the Royal Microscopical Society, to be held in the University Engineering Department, Trumpington Street, Cambridge, on Thursday 25th September 1997.
The colloquium is intended to cover recent progress made in the use of computers for the assessment, interpretation, restoration and enhancement of microscopical images, including, but not restricted to applications in optical and all kinds of electron microscopy. Of particular interest will be contributions concerning new approaches and techniques including 3D measurements and profiling, wavelets and fractals. It is hoped that papers will cover some of the comparatively new fields of application, including multi-channel imaging, instrumental control and remote microscopy. It is proposed also to organise a small exhibition of products and materials by local organisations involved in this area, as well as a visit to laboratories in the University engaged in this kind of work.
The colloquium takes place immediately after a three-day course, also entitled 'Computers in Microscopy', and it is intended that the two events be complementary. ( Please note that it is not necessary to attend the course in order to register for the colloquium.)
If you would like further information about the Colloquium and/or the Course please contact Rebecca Morden either by email: info-at-rms.org.uk, by telephone: +44 (0)1865 248768 or by fax: +44 (0)1865 791237.
Prospective contributors are invited to submit a synopsis of approximately 150-200 words, before 30 April 1997, to Dr D M Holburn, University Engineering Department, Trumpington Street, Cambridge CB2 1PZ, U.K. Enquiries may be made by electronic mail, to dmh-at-eng.cam.ac.uk, by facsimile to +44 1223 332662, or by telephone to +44 1223 332775.
******************************************************************************* Rebecca Morden Course Organiser Royal Microscopical Society, 37/38 St Clements, Oxford, OX4 1AJ, UK. Tel (0)1865 248768, Fax (0)1865 791237, email info-at-rms.org.uk *******************************************************************************
Thank you very much for providing information and experience in window breackage problem. I got windows replace service with the price of $6500 (does not including freight charges). We do not know how to handle this problem if the window does not stand in a reasonable lifetime or unlucky, got several window breackages. It must be a big meney gone! Is it possible to find a cheaper service? Any suggestion?
I have been tasked with bringing another technician into our SEM lab and would like to get info on the courses which are available for a person who is familiar with light microscopy but knows almost nothing about a SEM.
The desired course would be 5 days or less and be applicable to the level of knowledge described above. We deal with strictly materials analysis, so biological based courses are not applicable. The desired outcome would be someone who has a rudimentary knowledge of SEM theory and capable of taking basic SEM pictures and acquiring EDS spectra on non-challenging samples. We will be doing some training of the new tech, but due to heavy workloads, we can't devote the time to the training that we would normally do.
Geographically, I would like courses in the Southeastern USA, but will take info on courses anywhere in the US.
John Giles Senior Materials Engineer Honeywell Space Systems Clearwater, FL
} John Giles wrote:-----------------------------------------------. } } Hello Everyone, } } I have been tasked with bringing another technician into our SEM lab and would } like to get info on the courses which are available for a person who is } familiar with light microscopy but knows almost nothing about a SEM. } } The desired course would be 5 days or less and be applicable to the level of } knowledge described above. We deal with strictly materials analysis, so } biological based courses are not applicable. The desired outcome would be } someone who has a rudimentary knowledge of SEM theory and capable of taking } basic SEM pictures and acquiring EDS spectra on non-challenging samples. We } will be doing some training of the new tech, but due to heavy workloads, we } can't devote the time to the training that we would normally do. } } Geographically, I would like courses in the Southeastern USA, but will take } info on courses anywhere in the US. } } John Giles } Senior Materials Engineer } Honeywell Space Systems } Clearwater, FL
John:
A number of us run an SEM course at the University of Maryland called 'Practicle Aspects of SEM. This course is a 4 1/2 day course covering the basic theory and practice of SEM (including basic x-ray microanalysis) with direct hands-on experience for all students. In addition, students are encouraged to bring samples of their own. This year the courses are set for May 19-23 and May 26-30. If you would like more information please feel free to contact either me ( 315-859-4715) or Tim Maugel (301-405-6898)
Ken Bart
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323 USA kbart-at-hamilton.edu (315) 859-4715
We are in need of a vibration isolation pad for an Auger spectrometer, which has the dimensions of a standard SEM. Can anybody recommend a manufacturer or can recommend homemade solutions?
We have a WDS system for sale. The system consists of the computer, software, spectrometer controls and stage controls, NO DETECTORS or SPECTROMETERS are included.
It is a Kevex Delta 4 / Sesame system (type 8006) with a 5724-004 console, containing 4 spectrometers controls, 3 stage motors, and 4 spectrometer pre-amps and ratemeters, beam meter. A Tectronix plotter and a matrix printer a part of the package.
The system was configured to run on a JEOL JXA 8600 superprobe.
Price: $ 5000, you pay freight.
Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
I hope this doesn't seem out of palce on this list, but it is equipment I need to print spectra and images from ny TEM work. Has anyone had to replace the printhead on an Apple Color StyleWriter Pro and gotten a better price than the going Apple rate of $250! (new printer time?) Alternately, has anyone got a unit that was junked for other reasons that might have a usable printhead that they would be willing to sell/donate? The one that I use in my electron microscope lab had a malfunction which forced ink on top of the printhead and corroded out the contacts. Thanks in advance for any help/advice you can offer.
Sincerely, Andy Andy Buechele The Catholic University of America 409 Hannan Hall Washington, D.C. 20064 (202) 319-4995 FAX: (202) 319-4469
Y'all are an encyclopedia of knowledge, and I want to check out a volume... I have just recently been learning SEM and all the associated procedures that precede going on the scope. I have had to learn this all on my own, which is why I'm having this problem. When I do a cpd on a sample they always come out looking wrinkled. Am I going wrong in the cpd or is it some other step. I've heard that some people do a few purges of the system and some do a lot. I've just purged the ETOH until I see CO2 snowballs & then let it sit for 15 min. & then repeat the cycle 2 more times. Is this wrong? Can someone out there give me some pointers? I'm getting pretty tired of my stuff looking like raisins all the time. = (
Thanks in advance!
Paula = )
Paula Ssicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
This seems to be a question of general interest. As the manufacturer of the majority of ultrathin windows, MOXTEK can offer the following suggestions to improve window lifetimes.
1) Slow down the chamber venting. The most common field failure is what we call "bullet holes." These occur when particulate in the chamber is "swirled up" during venting and impacts the surface of the window.
2) Retract the detector before you vent, if you have this capability. (See #1 for the reason.) I saw an earlier suggestion to cover the detector with a plastic bag when it is not in use to minimize the exposure to this flying particulate. Good suggestion!
3) Never pump down your specimen chamber with a warm x-ray detector attached. Make sure you cool down the detector before pumpdown. Most of the detector systems have a molecular sieve or other getter material in the dewar vacuum to trap any residual gases. When you warm up the detector this material can outgas the trapped gases. If you then pump down the specimen chamber you reverse pressure the window. These windows are not designed to tolerate any back pressure.
4) Never overheat the window by exposure to a hot stage. Your instrument manufacturer should be able to recommend how to operate the system to minimize heat transfer to the detector window.
5) Consult with your EDX manufacturer for specific instructions for your system. Every detector/microscope combination is unique, and they should have some experience that will help you.
There are various thicknesses of windows available, but the above suggestions apply regardless of the thickness. If you continue to have failures with the thinnest windows, a thicker option is probably available through your EDX manufacturer.
(Please note that the Kevex B windows are unique to Kevex, so you should probably talk to them about the problem you had. With a silicon support grid like the MOXTEK ultrathin windows there is no problem in frequent pressure changes breaking the window.)
Hope this helps.
D. Clark Turner Director MOXTEK, Inc. 452 West 1260 North Orem, Utah 84057 phone (801) 225-0930 fax (801) 221-1121 email moxtek-at-moxtek.win.net
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REGARDING Job Posting
POSITION OPEN: Postdoctoral Scholar or Research Associate
Area: Materials Science/Analytical Electron Microscopy/Lorentz Imaging
Qualification: A PhD in Materials Science/Physics or related area. Very strong hands-on experience in various TEM techniques and their application to microstructural characterization at high resolution is required. Experience in Lorentz Imaging and TEM specimen preparation is desirable. The suitable candidate will be comfortable using the wide range of TEM instrumentation available at the NCEM. This post-doctoral fellow will spend a major fraction of the time working on the microstructural characterization of plastically deformed samples. The goal is to explore the correlation between the local physical/chemical microstructure in plastically deformed regions with local changes in magnetic structure(measured independently by SQUID imaging techniques). The remaining time will be devoted to the development of Lorentz Imaging techniques on our new Philips 200KV/FEG instrument using a variety of thin film samples.
The position is open immediately for at least one year, renewable upon mutual agreement for longer period, provided funds are available. Salary and benefits will commensurate with experience and skills.
For more information on the scientific/technical aspects of the position please contact :
Kannan Krishnan 72-209, NCEM Lawrence Berkeley National Laboratory Berkeley, CA 94720
Lehigh University, Bethlehem, PA has beginner as well as advanced SEM/EDS (+other) courses. These run for a week, and are well done. Sorry, I tossed my mailing about the course so I am missing the details. Believe it is scheduled for June. I am sure someone else will jump in with the details.
There are other (similar) courses available, but I have not experienced those....
Quantitative (even standardized) EDS analysis for carbon in Nb is difficult to impossible, no matter what the EDS manufacurers may imply. The same applies for carbon in Zr. Several things..... 1) Very light element (soft x-rays) in a high Z matrix. 2) Nb is a strong absorber of carbon x-rays. 3) Since the matrix carbon signal is weak, any surface carbon (contamination) can be a major contributor to the carbon peak. Together, these make for very large absorption correction coefficients and magnify surface contamination contributions. Translate that: error.
To maximize your chances....
Don't rely on standardless (semi) quant.
A clean, high vacuum is required to minimize surface (carbon) during analysis.
Specimen surface preparation is crucial. A flat, well polished, and very clean specimen surface must be achieved.
Do not coat the specimen (NbC is sufficiently conductive anyway).
Minimize analysis volume and depth. - (A) Use as low a beam voltage as practical. (B) High tilt angles will skew the volume to near surface. Caveats: (A) Nb, L-line (family) response less predictable than Ka line. I have run across some quant software which won't use L-lines- at least for stdless quant. (B) Less than perfect quant compensation for high tilt may add some error...(especially for stdless).
For stdless quant, I have seen demos produce errors } 300 percent.
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I am trying to do trace Carbon analysis in Nb material. The Be window of my EDX can be opened in order to detect C, N, O, but the quantification seems impossible. Anybody has similar experience?
Message-Id: {2.2.32.19970308041320.006ac864-at-pop.unixg.ubc.ca} X-Sender: mager-at-pop.unixg.ubc.ca X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Dear Hasso, I have had some success reducing the vibration evident on my SEM by making four pads from Sorbathane. I used a fairly stiff number of Sorbathane about 3/8 inch thick and cut four pads about four inches square. The shop made me eight steel plates to go on either side of the Sorbathane and I put these under the corners of the column section. The vibration on my SEM (from being on the fourth floor) went down about half. It may not be enough for Auger but it is inexpensive. TMC, who advertises in most JMSA journals, has a full set of platforms. Phone:508-532-6330, fax:508-531-8682. I have no experience with their service. You wrote:
} We are in need of a vibration isolation pad for an Auger spectrometer, } which has the dimensions of a standard SEM. Can anybody recommend a } manufacturer or can recommend homemade solutions? } Best of luck, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
} Y'all are an encyclopedia of knowledge, and I want to check out a } volume... I have just recently been learning SEM and all the associated } procedures that precede going on the scope. I have had to learn this all } on my own, which is why I'm having this problem. } When I do a cpd on a sample they always come out looking wrinkled. } Am I going wrong in the cpd or is it some other step. I've heard that some } people do a few purges of the system and some do a lot. I've just purged } the ETOH until I see CO2 snowballs & then let it sit for 15 min. & then } repeat the cycle 2 more times. Is this wrong? } Can someone out there give me some pointers? I'm getting pretty } tired of my stuff looking like raisins all the time. = ( } } } Thanks in advance! } } Paula = ) } } Paula Ssicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu
The main problem particularly newcomers to CPD have is trying to do everything too quickly. I'm no expert at CPD having only done it a few times, but I have been told:
a) when flushing, it need to be done SLOWLY - if the CO2 boils (or even simmers gently), then the specimen will be wrecked.
b) It is important to leave your specimen in the liquid CO2 for long enough for proper infiltration, and that this needs doing at least two or three times to make sure all the acetone/alcohol is displaced - even for small specimens (1mm), this can take 3-4 hours. Larger specimens might require 24 hrs.
c) At the end, when releasing the CO2 gas, again it must be done slowly - 15-20 mins. Sudden pressure releases will blow the specimen apart.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
Thanks to all who responded to my question about teaching supplies. I received about 20 responses. They fell into three basic types 1) give each student an allotment at the beginning of the semester and require them to buy more if they use it up; 2) charge each student a flat fee and let them use at will; 3) require them to but their own supplies at a local camera store. Some labs separated EM film (which can be hard to purchase locally) from paper (which is stocked locally).
I'm leaning towards using options #1 or #3.
Thanks again.
Bob
Robert R. Wise Plant Physiologist and Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
You should look into Neurolucida by MicroBrightField. It is a 3D morphometric analysis system. MBF also is also active in stereology analysis and was an exhibitor at the Soc. Neurosci meeting. You can get more info from their web site at www.microbrightfield.com/microb/ or via email at info-at-microbrightfield.com.
On Thu, 27 Feb 1997, Cheri Owen wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } A friend of a friend is looking for a morphometric system with Scope, } stylus, PC etc. They saw some at the recent Neurosci meetings, but didn't } keep the brochures. Any info would be greatly appreciated. } } Thanks } } Cheri Owen } Detroit Neurotrauma Institute } Wayne State University } Detroit, MI } } }
Edmund Glaser, D. Eng. Dept. Physiol. Univ. Md. School. Med. Baltimore, MD 21201 USA Ph: (410) 706-5041 Fax: (410) 706-8341
We are trying to photograph bacteria using phase contrast optics. We are having trouble getting a thin enough layer to obtain 'fully focused" micrographs at 100x under oil. We are using a fixed suspension mixed with "aquatex" mounting medium. Any suggestions? Thanks David Wild
I am sorry that this is a little late. I will be trying to get this message out to the individual authors who will be presenting who do not receive it through the Listserver. If you are the principal author or a coauthor on a paper being presented at Symposium Z at the Spring '97 MRS meeting, please let me know that you received this anouncement ASAP.
The papers that you are submitting to the "Workshop on TEM Specimen Preparation-IV", Symposium Z should be tutorial in nature. They should be written in a "how I did it and here's how you can do it" mode with all the tidbits and tricks-of-the-trade that make your technique work. Follow the authors guidline for the paper for the MRS proceedings except for length. Use as many pages as you need to tell your story. Use as many pictures and diagrams that you need to get the information across. If you have any questions, contact me or Ron Anderson.
- -Scott Walck
Ron Anderson, Chair IBM, ZIP E-70 East Fishkill Facility Hopewell Jct., NY 12533 *(914)892-2225 (-2003 FAX) ron-anderson-at-vnet.ibm.com
Scott D. Walck, Co-Chair Materials Directorate, Wright Laboratory Wright Patterson AFB, OH 45433-7750 *(513)255-5791 (-2176 FAX) walcksd-at-ml.wpafb.af.mil
Can anyone please tell me the name of staining books in the field of Immuno-bed tissues as well as the companies from which I can order it. We are interested in - Histology and Pathology - Immunofluorescence - Immunohistochemistry - Histofluorescence - Enzymehistochemistry
Thanks Hildagonda van der Merwe
University of Pretoria Faculty of Veterinary Science Dept. of Pathology Onderstepoort 0110
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello All, } } } Y'all are an encyclopedia of knowledge, and I want to check out a } volume... I have just recently been learning SEM and all the associated } procedures that precede going on the scope. I have had to learn this all } on my own, which is why I'm having this problem. } When I do a cpd on a sample they always come out looking wrinkled. } Am I going wrong in the cpd or is it some other step. I've heard that some } people do a few purges of the system and some do a lot. I've just purged } the ETOH until I see CO2 snowballs & then let it sit for 15 min. & then } repeat the cycle 2 more times. Is this wrong? } Can someone out there give me some pointers? I'm getting pretty } tired of my stuff looking like raisins all the time. = ( } } } Thanks in advance! } } Paula = ) } } Paula Ssicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu
Paula, It sounds like your samples are still "wet". Make sure that your are dehydrating your samples long enough. The ETOH or acetone must be 100% pure. The time you are alloting in the CPD is too short. This will also leave your sample wet with acetone or ETOH. When you drain the ETOH do it slowly and place a dry Kimwipe close and infront of the vent. When it no longer gets wet after a few purges the ETOH is gone. Then Let it sit for 30 min. more. Bring the temp. of the chamber up to about 37C slowly. Vent the CO2 over 10 min until the pressure is zero. The above is all sample dependent and must be worked out by trial and error. Plant material will take longer than a cell cultuer on a cover slip.
Greg Rudomen University Microscopy Imaging Center S.U.N.Y. Stony Brook Greg-at-umic.sunysb.edu
This is to remind everyone that extended abstracts for Microscopy & Microanalysis '97 need to be subimtted by MARCH 15! That is this Saturday! Papers received after that date will not be accepted.
If you have misplaced or have not received a bulletin or need more information, you can contact
Microscopy & Microanalysis '97 4 Barlows Landing Rd., Suite 8 Pocasset, MA 02559 phone: 508-563-1155 Toll-free: 800-538-3672 FAX: 508-563-1211
Email: BusinessOffice-at-MSA.Microscopy.com or WWW: http://www.MSA.Microscopy.com
Meeting information is also available at http://www.bright.net/~strecker/msno/mm97.html
Poster instructions can be found at the MRS web site, www.mrs.org
You should have recieved author instructions. Please contact MRS if you have not. The Materials Research Society, 9800 McKnight Road, Pittsburgh, PA 15237-6006 Phone: 412/367-3004, Fax: 412/367-4373, Email: info-at-mrs.org
If you haven't, this should get you started.
For camera-ready text, on an 8-1/2 x 11 inch paper, the margins are left: 1 inches right 1 inches top: 1/2 inch bottom: 1 inch Page number at 1/2 inch from bottom of page.
That gives 6.5 inch wide x 10 inch box to put your text, single spaced, 12 point font Times Roman. Use the AIP reference style or the MRS refernce style. The manuscripts will be reduced by 26%. Scale markers on all micrographs. Check out other proceedings for style if you haven't received your instructions.
Bring the original manuscript and (3) photocopies to the meeting. You will need to have the MRS copyright form filled out. Check the bulletin boards on where to take your paper. Ron or myself will be collecting them from you and will post an announcement.
The Royal Microscopical Society Computers in Microscopy ,September 1997, Cambridge CALL FOR PAPERS
A Colloquium on various aspects of the application of computers to microscopes and microscopical techniques is being planned by the Royal Microscopical Society, to be held in the University Engineering Department, Trumpington Street, Cambridge, on Thursday 25th September 1997.
The colloquium is intended to cover recent progress made in the use of computers for the assessment, interpretation, restoration and enhancement of microscopical images, including, but not restricted to applications in optical and all kinds of electron microscopy. Of particular interest will be contributions concerning new approaches and techniques including 3D measurements and profiling, wavelets and fractals. It is hoped that papers will cover some of the comparatively new fields of application, including multi-channel imaging, instrumental control and remote microscopy. It is proposed also to organise a small exhibition of products and materials by local organisations involved in this area, as well as a visit to laboratories in the University engaged in this kind of work.
The colloquium takes place immediately after a three-day course, also entitled 'Computers in Microscopy', and it is intended that the two events be complementary. ( Please note that it is not necessary to attend the course in order to register for the colloquium.)
If you would like further information about the Colloquium and/or the Course please contact Rebecca Morden either by email: info-at-rms.org.uk, by telephone: +44 (0)1865 248768 or by fax: +44 (0)1865 791237.
Prospective contributors are invited to submit a synopsis of approximately 150-200 words, before 30 April 1997, to Dr D M Holburn, University Engineering Department, Trumpington Street, Cambridge CB2 1PZ, U.K. Enquiries may be made by electronic mail, to dmh-at-eng.cam.ac.uk, by facsimile to +44 1223 332662, or by telephone to +44 1223 332775.
******************************************************************************* Rebecca Morden Course Organiser Royal Microscopical Society, 37/38 St Clements, Oxford, OX4 1AJ, UK. Tel (0)1865 248768, Fax (0)1865 791237, email info-at-rms.org.uk *******************************************************************************
Location : Carolinas HealthCare System Charlotte, North Carolina, USA
Shift: Full-time, M-F, 8a-4:30p.
Essentials: Fix, dehydrate, and embed biological tissue specimen for transmission e.m. Thick and thin sectioning, Basic operation of Philips CM10 electron microscope. Develop and print photomicrographs. Maintain logs and records of work,
Requirements: BA/BS degree in related area. Minimum 1.5 year relatively recent experience in electron microscopy, preferably in a work setting. Good "people skills."
} We are in need of a vibration isolation pad for an Auger spectrometer, } which has the dimensions of a standard SEM. Can anybody recommend a } manufacturer or can recommend homemade solutions?
There are some low cost pneumatic mounts available from Barry Controls in a wide range of load handling capacities. The trade name is Stabl-Levl. I used these to successfully isolate a heavy FTIR unit that was sensitive to vibration. Unless you can attach them directly to your instrument, however, you will still need a stiff plate to set it on with the pneumatic mounts between the plate and the floor. The address and phone # I have for Barry Controls is: 700 Pleasant Street Watertown, Mass. 02172 617-923-1150 Haven't ordered from them in a few years, so you might need to check it if you can't connect. Disclaimer: I have no interest in Barry Controls other than being a satisfied customer.
Andy Buechele The Catholic University of America 409 Hannan Hall Washington, D.C. 20064 (202) 319-4995 FAX: (202) 319-4469
Our Auger with scanning tunnel microscope (with console)hangs from ceiling beams on three springs if you need details you may contact Prof. Achete (achete-at-metalmat.ufrj.br) in this department Best regards Prof.Walter A.Mannheimer Metallurgy and Materials Engineering Federal University of Rio de Janeiro POBox 68505 21945 Rio de Janeiro, Brazil Voice +5521 280-7443 (Dept.office) +5521 590-0579 (direct) Fax +5521 290-6626 Email: wamann-at-metalmat.ufrj.br
Can anyone tell me the best way to mount a sample for hot stage work. We have a cross section of a silicon nitride sample that we want to examine at elevated temperatures (up to about 1000 deg C, if our hot stage will get us there!) The sample is a cross section made by tripod polishing and it must be mounted on a Mo or Ta washer for insertion into the scope, how are we going to "glue" it to the ring so that the "glue" wont decay and the sample fall off or move about at high temp? What we need is a very low vapor pressure adhesive that can withstand high temperatures!
Any help appreciated. Thanks.
Jfm.
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313) 936-3352 FAX (313) 936-3352 Cellular Phone: (313) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Due to a projected streamlining of the review and production process, abstracts for the June meeting of the Microscopy Society of Canada can be accepted up to 2 weeks later than previously advertised, i.e. they must be received in Edmonton by 1 April.
The deadline for conference pre-registration is 1 May 1997, as generally advertised (but NOT 1 March as stated on the registration form!)
Ray Egerton, Physics Dept, University of Alberta, Edmonton, Canada T6G 2J1 Phone: 403-492-5095, FAX: 403-492-0714, e-mail: egerton-at-phys.ualberta.ca ------------------------------------------------------------------------
Afew weeks ago, there was someone who was doing a survey of the best types of gloves to use in an EM lab. Of course I didn't save the information passed on, and of course now we're interested in those results here in our lab. If anyone has those results, please let me know. Also, we work with a lot of the low-temperature resins (Lowicryls, LR Gold), if anyone knows of the best gloves to work with that stuff let me know that too.
Thanks oodles!
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I would like tofind someone in the S.F. Bay area that has a freeze-fracture machine that would be available for doing samples. I have a colleague in the area who is looking for a facility where she might be able to do some samples. We would be very grateful for any leads. Thanks in advance. ML
Mei Lie Wong Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 email wong-at-msg.ucsf.edu
I am about to replace the sputter ion pump elements in our two JEOL TEM's. JEOL used to be able to recondition the elements used in their 2000FX microscopes but no longer provide this service. Can anyone suggest another company that does this sort of work? How long can I expect a reconditioned unit to last compared with a new one and what sort of cost would be reasonable given that JEOL are charging $2.5k and $4.5k (Australian dollars) for new units? Thanks in advance for any advice you can give.
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.
The Oklahoma Microscopy Society (OMS) will be hosting its 19th Annual Spring Workshop on Friday, April 4, 1997.
Topic: FORENSIC MICROSCOPY
MORNING SESSION: 129 George Cross Hall, University of Oklahoma in Norman, OK.
8 - 9 am: Registration
9 - 9:05: Greetings
9:05 - 9:50: Guest speaker: Mark Betts (from Oxford Instruments) will speak on "Microanalysis in the Forensic Laboratory"
9:50 - 10: Break
10 - 12: Guest speaker: Keith Ferrell (from the Oklahoma State Bureau of Investigation (OSBI)) will speak on "Overview of Forensic Microscopy"
12 - 1:30pm: Lunch/Executive Meeting/Open Meeting
AFTERNOON SESSION: OSBI Central Laboratory, 2132 N.E. 36th St. in Oklahoma City
1:30 - 2: Travel to the OSBI
2 - on: Tours of the OSBI Facility
REGISTRATION (includes lunch):
You can download a copy of the registration form by visiting the OMS website at http://www.ou.edu/research/electron/oms/ maintained by Dr. Scott Russell, Nobel Electron Microscopy Lab at the University of Oklahoma.
OMS members: the Newsletter was sent out Monday, March 10, 1997 so you should be receiving it this week. A registration form/envelope has been included in the newsletter. Please return it as soon as possible so as to estimate numbers for lunch and parking permits.
Student and Professional Members: $5 Student Non-Members: $10 Professional Non-Members: $15
Pre-register until April 2, 1997 with Phoebe Doss OMS President Elect Dept. of Plant Pathology 110 NRC Oklahoma State University Stillwater, OK 74078 (405) 744-7995 Email: pjdoss-at-okway.okstate.edu
Your question, "how to stain enzymes for EM" is so broad as to be unanswerable, I think. Which enzymes?, Plant tissue or animal? Cells grown in culture? I am sure someone on the list can help if your question is more specific.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
On 03/10/97 17:21:17 you wrote: } I would like tofind someone in the S.F. Bay area that has a freeze-fracture } machine that would be available for doing samples. I have a colleague in } the area who is looking for a facility where she might be able to do some } samples. } We would be very grateful for any leads. Thanks in advance. ML } } Mei Lie Wong } Department of Biochemistry } HHMI-UCSF } Ph. 415-476-4441 Fax 415-476-1902 } email wong-at-msg.ucsf.edu
Dear Mei, Please call Nancy Smith at Cal State Hayward (main number 510-885-3000). I am sure she cal help you. Regards
Igor C. Ivanov, SIMS,Auger,TEM,SEM Senior Scientist Analytical Services RJ LeeGroup, Inc. Contract Research 530 McCormick Str. (510)-567-0480 phone San Leandro, CA 94577 (510)-567-0488 FAX
} Return-Path: {das3-at-Lehigh.EDU} } X-Sender: das3-at-mail.Lehigh.EDU } Date: Tue, 11 Mar 1997 16:26:22 +0100 } To: Donna Jacobs {jacobs-at-uimrl7.mrl.uiuc.edu} } From: das3-at-Lehigh.EDU (David A. Smith) } Subject: Re: Open Position - Research Electron Microscopist } } DR. DAVID SMITH PASSED AWAY SEPTEMBER 1996. PLEASE REMOVE FROM ALL E-MAIL } AND MAIL LISTINGS. THANK YOU. } } MAXINE MATTIE } } David A. Smith, Department of Materials Science and Engineering, Lehigh } University, 5. E. Packer Ave., Bethelehem, PA 18015, Ph 610 758 4231, Fax } 610 758 4244 } } } Donna Jacobs MRL Administration University of Illinois 104 South Goodwin Avenue Urbana, Illinois 61801 Phone (217) 244-2944 Fax (217) 244-2946 email - jacobs-at-uimrl7.mrl.uiuc.edu
Good Afternoon (Central Std Time USA) all Microscopists:
I have just had a request to determine the refractive index of zinc pyrithione, a crystalline, colorless, transparent powder. Does anyone know what the RI is? It's not in McCrone Atlas. I will be getting a sample Monday and with a set of Cargille RI liquids, I will determine the RI.
Alternatively, I have a B & L Abbe type refractometer used to determine refractive index of liquids. However, I thought I read somewhere that it could be used to determine the RI of solids. If this is in fact possible, does anyone out there know how?
Thanks to all who responded to my cpd inquiry. The consensus seems to be that fresh, dry 100% ETOH is very important and that I need to keep my samples longer in the liquid CO2 to get a better exchange with the ETOH.
I'll try out all the suggestions. Hopefully, the only raisins I'll get from now on will be in my breakfast cereal. = )
Thanks again.
Paula = )
p.s. I'm going to phone some vendors of the resins & such to see if they've done studies as to which type of gloves work best. I'll keep you posted.
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I am acquiring an ISI SX-30E and need a copy of the manual for this instrument without paying the hundreds of dollars Topcon wants for such a manual. Any suggestions where I might be able to get a xerox of the manual or borrow a copy to xerox. Any help would be appreciated.
Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) advertise a service for rebuilding sputter ion pumps (p. 25 of their catalog), and state that a properly rebuilt pump should "have the same performance as a new pump". The cost of rebuilding varies from about $400 to about $3000, depending on the size and type of pump No commercial interest - just trying to be helpful.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Does anyone who performs vascular casting know where to obtain a product called Mercox CL2? A fellow electron microscopist has heard that it is the latest thing in vascular casting when mixed with methyl methacrylate but hasn't been able to get any further info. If anyone knows who makes it and where it can be bought we would like to hear.
Thanks in advance
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
For the theory you need to look up Snell's Law and Molecular refractivity. The RI relates to the velocity of light through various substances. The difference between air and water RI results in a stick appearing angled at the interface. Transparent specimen become invisible if immersed in a liquid of identical RI. This is extensively used in police scientific work were glass fragments can be identified to come from a crime scene. Very simple, put a sample onto a microscope slide. Add a drop of RI solution. If the submersed specimen becomes invisible under the microscope, than the refractive index of the specimen is that of the RI test solutions. Some experimenting to find the right solution is required. Good luck. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au ---------------------------------------------------------------------------- ----- } From: Damian_Neuberger_at_RLT013-at-ccmailgw.mcgawpark.baxter.com } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Refractive Index } Date: Wednesday, 12 March 1997 6:49 } } I have just had a request to determine the refractive index of zinc } pyrithione, a crystalline, colorless, transparent powder. Does anyone } know what the RI is? It's not in McCrone Atlas. I will be getting a } sample Monday and with a set of Cargille RI liquids, I will determine } the RI.
} Transparent specimen become invisible if immersed in a liquid of identical } RI. This is extensively used in police scientific work were glass fragments } can be identified to come from a crime scene. Very simple, put a sample } onto a microscope slide. Add a drop of RI solution. If the submersed } specimen becomes invisible under the microscope, than the refractive index } of the specimen is that of the RI test solutions.
Unfortunately it is not quite that simple. If your material is a glass and/or has one refractive index this procedure will work fine. Don't forget to correct for temperature and wavelength!
But-- If your sample has two or more refractive indexes the procedure becomes a lot more complicated. You need to separate one refractive index from another and this can be done with the polarized light microscope with a lot of practice. I suggest "Handbook of Chemical Microscopy, Vol One" by Chamot and Mason, if you can find a copy, as a reference. The procedures and techniques are too detailed for a short note.
Best wishes.... Frank
PS i was unable to find any optical data in Winchell's "The Optical Properties of Organic Compounds"
It is possible to determine the RI of SOME solids with the B&L Abbe refractometer. But I don't know if it is possible to do this on a powder; I have never done it. The more critical requirements for doing a solid (if you are still interested) are:
1) You must have very intimate contact between the solid and the lower prism. This is normally accomplished by polishing the sample such that it is extremely flat, and then a contact fluid (1-bromonapthalene) is used to form the intimate contact. Use as little contact fluid as necessary (usually a very small drop).
2) It is important to have a 90 degree angle between the bottom prism and one edge of your sample. This edge also needs to be highly polished, like the contact surface.
Good luck with the powder; if anyone suggests a way to do this with the refractometer, I would also be interested.
Regards,
-Bob ************************************ Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909) 399-1311 email: Bob_Citron-at-cc.chiron.com ************************************
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Good Afternoon (Central Std Time USA) all Microscopists:
I have just had a request to determine the refractive index of zinc pyrithione, a crystalline, colorless, transparent powder. Does anyone know what the RI is? It's not in McCrone Atlas. I will be getting a sample Monday and with a set of Cargille RI liquids, I will determine the RI.
Alternatively, I have a B & L Abbe type refractometer used to determine refractive index of liquids. However, I thought I read somewhere that it could be used to determine the RI of solids. If this is in fact possible, does anyone out there know how?
Osmolarity is an easily measurable characteristic, by e.g. freezing point depression. In simple terms it is the total concentration of solutes (including ions) in a solution, and this does not depend on whether the solutes can cross cell membranes or not. Tonicity relates to the osmotic GRADIENT due to solutes that affects a semi-permeable membrane ( i.e. a membrane that is permeable to water); ONLY solutes that do not cross the membrane contribute to this effect. The two characteristics are different and obviously have vastly different consequences on membranes. Weak bases, even though ionized, have some measurable permeability and so their contribution to tonicity must be much less than their contribution to osmolarity. Osmium is hydrophobic and quite permeable through membranes, so contributes *nothing* to an osmotic gradient that can disrupt the membranes (even assuming it doesn't alter their permeability to tonic agents like sucrose). So osmium contributes nothing to tonicity, but certainly does contribute to osmolarity.
I personally wonder how to express the tonicity of compounds that are somewhat permeable, such as weak bases or short carboxilic acids & aldehydes. I guess the concept of tonicity either doesn't apply or is operational in such cases, depending on the time scale of interest. Can anyone comment or give a reference to a good detailed textbook? Something about reflection coefficients?
Richard
} } } Gary Dietrich Chinga {garyc-at-stud.ntnu.no} 03/12/97 09:31am } } } Hi!
Can someone explain the relation between the tonicity and osmolarity of a fixation solution?
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Gary,
Tonicity is a relative, unitless comparison of one solution to another in which the first solution (presumably the water in your tissue sample) is hypertonic, hypotonic or isotonic to the second solution (presumably your fixative). Osmolarity is an absolute scale (usually in some type of pressure unit) which describes the concentration of osmolytes in a single solution. So you need to know the osmolarity of your tissue so you can set the osmolarity of your fixative to be isotonic with respect to the tissue.
Cheers
Bob
Robert R. Wise Plant Physiologist and Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
I have a user trying to follow a new protocol which uses "an antistatic solution of ... Denkil-SEM (Hodogaya Chemical)" but I have no idea what "Denkil-SEM" is can anyone out there give us a hint? (Before we start trying other antistatic solutions, i.e. Static Guard, or Cling Free!).
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
I have a question which has undoubtedly been answered before...! We are going to buy a flat bed scanner, probably a Microtek with a transparency adapter. We would primarily use it for scanning in TEM and SEM (polaroid) negatives. Should we go for the 24-bit color, 300 x 600 dpi version, or the more expensive (ouch) 30 bit color, 600 x 1200 dpi version? Our resident computer guru says that we should go with the cheaper version because we are unlikely to have an output source with better than 600 dpi for most stuff. It's true; the printers around here are mostly 300 - 600 dpi. Is there a compelling reason for the better model? Will I be really sorry in a year if I don't?
Thanks you all for all the expertise and advice on so many different topics!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I've been checking around on the glove issue. It seems that there is no one glove that works for everything (darn!). Usually if it's good for one thing, it's lousy for another. We work with Lowicryl resins a lot and the best glove to use is........
Nitrile, those blue smelly things that don't stretch worth a darn.
I also received some info. from a paper (thanks, Bill = ) ), that told of an actual test done with various resins & such on different makes of gloves. The paper is "Glove Material For Handling Epoxy Resins" by David L. Ringo, Douglas R. Read and Eugene H. Cote-Robles in the Journal of Electron Microscopy Techniques 1:417-418 (1984). Pretty interesting stuff!
Safety in the lab is an important thing. So each lab must make it's own mind as to which type of glove they want to use. The important thing is that you get your students to glove up and work in the fume hoods and be aware that almost all of the chemicals in an EM lab are hazardous.
Happy Scoping!
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
********************************************* * Metropolitan Microscopy Society Meeting * *********************************************
Date: Wednesday, March 26, 1997
Time: 10:00 AM
Place: Howard Johnson Lodge, 393 Route 17, Paramus, NJ
Directions: The Howard Johnson Lodge is on the southbound lane of Route 17. From the Garden State Parkway, use Exit 163 if you are northbound, and Exit 165 if you are southbound.
10:00 - 10:30 am Registration ($5.00), Coffee and Danish.
10:30 - 10:45 am Introductory remarks and society announcements -- Philip Flaitz.
10:45 - 11:30 am HIGH RESOLUTION LOW VOLTAGE SCANNING ELECTRON MICROSCOPY, Dr. Frederic Cosandey, Dept. of Ceramic and Materials Science, Rutgers, The State University of New Jersey, Piscataway, NJ.
Scanning Electron Microscopy at low voltages (0.2 - 5 keV) has many advantages over more conventional electron energies such as reduced electron range, increased secondary electron yield, reduced radiation damage and charging artifacts. However, in order to maintain high resolution at the lowest voltages, special lens designs are required to correct for the chromatic aberration of the probe forming objective lens. In addition, improved secondary electron detection systems must be implemented. In this talk, the relative merits of various objective lens designs will be discussed with special emphasis on the electrostatic lens with retarding field concept. Also, the unique advantages of low voltage SEM for secondary and backscattered electron imaging modes will be discussed. A few specific examples will be presented including domain size measurements in block co-polymers, oxide surface structure determination and phase identification in nanocomposites.
11:30 - 12:15 pm MICROBEAM ANALYSIS OF POLYMERS, DRUGS AND INOR- GANIC MOLECULES BY INFRARED MICROSPECTROSCOPY, Dr. John A. Reffner, Spectra-Tech Inc., Shelton, CT 06484
Infrared microspectroscopy (IMS) is a growing technology for microbeam analysis of organic and molecular materials. As the fusion of scanning electron microscopy with x-ray emission spectroscopy created an exciting way for microscopists to investigate elemental composition, so has IMS impacted the way we now investigate the molecular chemistry of materials. IMS is essentially a photon probe --- producing spectral data by the absorption or reflection of infrared radiation. Applications to polymers, pharmaceuticals, electronics and forensics illustrate the wide range of uses of IMS. In reviewing IMS technology, both the strengths and limitations of current instruments are presented with a look at directions for future developments.
12:30 - 1:30 pm Lunch.
For additional information, please contact:
Phil Flaitz, IBM Analytical Services pflaitz-at-vnet.ibm.com, 914-892-3094
I know this question has been asked here before, so bear with me. Where can I get one of those red antistatic guns, now that they are no longer handled by Fullam? Their representative told me that there is supposedly a hefty supply of these units still in the U.K., and that Fullam doesn't handle them anymore because they'd have to buy more than they could sell. I know Discwasher used to distribute them. Does anyone know of the English company that has this huge supply? Are there any distributors? Is there any similar product? Appreciate any leads.
} Can someone explain the relation between the tonicity and osmolarity } of a fixation solution?
= = = = = = = = = = = = = = = =
Osmolarity is a measure of one of the colligative properties (osmotic pressure) of a solution. In a rough sense it is the measure of the amount of solute in a solution, but the degree of dissociation of the solute affects its osmolarity, so osmolarity is not necessarily a direct measure of molar concentration. (For example, a 1 molar solution of NaCl will have *approximately* twice the osmotic pressure [osmolarity] of a 1 molar solution of sucrose, since the NaCl dissociates into 1 molar Na+ and 1 molar Cl-.)
Two solutions (not necessarily of the same solute or of a single solute) are isosmotic if they have the same osmolarity (osmotic pressure). If two isosmotic solutions are placed on opposite sides of a semipermeable membrane, the osmotic pressure on each side is the same and there is no *net* movement of water across the membrane. However, if the osmolarity of the two solutions is not the same, then water will move across the membrane from the hypo-osmotic solution (more dilute) to the hyperosmotic solution (less dilute) until the osmotic pressure on each side is equal (a number of other factors affect this as well, but let's ignore them here).
Tonicity refers to the response of *cells or tissues* to the solutions in which they are immersed. If cells are placed in a hypertonic solution, net movement of water will be out of the cell, causing the cell to shrivel. If cells are placed in a hypotonic solution, net movement of water will be into the cell, causing the cell to swell or burst. Tonicity is useful only in reference to a particular cell or tissue.
Thus, the microscopist wishes to add sufficient solute(s) to the fixation solution so that the solution has the correct osmolarity (measured in milliosmols) so that it will have the desired tonicity with respect to the cells that are being exposed to the fixative. [There is debate whether the solution should be slightly hypertonic or hypotonic, but then that is another subject about which we may choose to debate.]
To summarize, osmolarity is a measurement of solute concentration (measurement can be made in a beaker). Tonicity is a comparison of osmolarities between a cell and the solution to which it is exposed.
I hope that this does not leave readers more confused than they were before. }
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
The 30 bits will give you additional shadow and highlight detail that 24 bits will not provide. Now, you may ask 'my software can only handle 24 bits so what good does 30 bits do when I will have to convert it down?'. When you convert the file to 24 bits, the system will choose from the best 24 bits to produce the converted file. I would equate it to a survey or poll - the larger your sample, the more accurate your results.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation
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I have a question which has undoubtedly been answered before...! We are going to buy a flat bed scanner, probably a Microtek with a transparency adapter. We would primarily use it for scanning in TEM and SEM (polaroid) negatives. Should we go for the 24-bit color, 300 x 600 dpi version, or the more expensive (ouch) 30 bit color, 600 x 1200 dpi version? Our resident computer guru says that we should go with the cheaper version because we are unlikely to have an output source with better than 600 dpi for most stuff. It's true; the printers around here are mostly 300 - 600 dpi. Is there a compelling reason for the better model? Will I be really sorry in a year if I don't?
Thanks you all for all the expertise and advice on so many different topics!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I would suggest to check out the speed as well as the resolution. We have an old HP-IIcx here that easily beats a newer cheaper scanner hands down. The software is also not as convenient as HP's deskscan package.
Regarding resolution, remember it will take a few of those printer pixels to dither up the gray scale for each of your digitized pixels. Thus, a 600 dpi printer may only be able to handle about a 200 dpi image. Of course if you are scanning 35 mm slides and enlarging them, you will need the high res scanning.
At 11:26 AM 3/12/97 -1000, Tina wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Donald Lovett's explanation quoted below is very good except that it perpetuates the myth that tonicity is related to the difference between two OSMOLARITIES on opposite sides of the membrane. That is ONLY true if the membrane involved is impermeable to ALL of the solutes contributing to the osmolarity. Since formaldehyde is soluble in both benzene and chloroform I expect it to be quite permeable across cell membranes. Thus it will contribute to osmolarity, but should contribute little to tonicity. The tonicity of your fixative should be adjusted with other components, and the final osmolarity of an isotonic fixative will be approx 300 mOsm PLUS the value of osmolarity contributed by the formaldehyde present.
To reiterate, IF a compound is permeable across the membrane it contributes to osmolarity but does not contribute to the osmotic GRADIENT. Thus it does NOT contribute to tonicity.
Caveats: Practically speaking, the rate at which a permeable solute crosses the membrane will affect the transient forces on the membrane. I assume tonicity is ill-defined unless the system is at equilibrium. The situation becomes even more complicated if the compounds added (such as aldehydes or alcohols) alter the permeability of the membrane for other buffer components present. And if differences in rates of diffusion through a chunk of tissue are involved, are we really still talking about tonicity?? That calls for empirical, not theoretical, optimization of the recipe used !!
On that note, how many of you microscopists have actually compared e.g. confocal images of cell cultures fixed with buffers of different tonicity with equivalent images of live cultured cells? The time to diffuse through a monolayer of cells should be negligible. What do you find is optimum?
Richard
} } } Donald Lovett {lovett-at-tcnj.edu} 03/12/97 05:03pm } } } Osmolarity is a measure of one of the colligative properties (osmotic pressure) of a solution. In a rough sense it is the measure of the amount of solute in a solution, but the degree of dissociation of the solute affects its osmolarity, so osmolarity is not necessarily a direct measure of molar concentration. (For example, a 1 molar solution of NaCl will have *approximately* twice the osmotic pressure [osmolarity] of a 1 molar solution of sucrose, since the NaCl dissociates into 1 molar Na+ and 1 molar Cl-.)
Two solutions (not necessarily of the same solute or of a single solute) are isosmotic if they have the same osmolarity (osmotic pressure). If two isosmotic solutions are placed on opposite sides of a semipermeable membrane, the osmotic pressure on each side is the same and there is no *net* movement of water across the membrane. However, if the osmolarity of the two solutions is not the same, then water will move across the membrane from the hypo-osmotic solution (more dilute) to the hyperosmotic solution (less dilute) until the osmotic pressure on each side is equal (a number of other factors affect this as well, but let's ignore them here).
Tonicity refers to the response of *cells or tissues* to the solutions in which they are immersed. If cells are placed in a hypertonic solution, net movement of water will be out of the cell, causing the cell to shrivel. If cells are placed in a hypotonic solution, net movement of water will be into the cell, causing the cell to swell or burst. Tonicity is useful only in reference to a particular cell or tissue.
Thus, the microscopist wishes to add sufficient solute(s) to the fixation solution so that the solution has the correct osmolarity (measured in milliosmols) so that it will have the desired tonicity with respect to the cells that are being exposed to the fixative. [There is debate whether the solution should be slightly hypertonic or hypotonic, but then that is another subject about which we may choose to debate.]
To summarize, osmolarity is a measurement of solute concentration (measurement can be made in a beaker). Tonicity is a comparison of osmolarities between a cell and the solution to which it is exposed.
I hope that this does not leave readers more confused than they were before. }
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
I'd like to forward this announcment of an open post-doc position to the list:
} The Laboratoire d'Analyse des Materiaux (LAM), of the "Centre de Recherche } Public-Centre Universitaire" in Luxembourg, has an immediate opening for a } two years post-doc position. } The core of the subject to be covered deals with analytical TEM (EELS and } EDX) : sensitivity, quantification, artifacts,... on complex samples. } As a side subject, this person will have to help in optimizing the } preparation of cross sectional samples: ion dimpling/microtome sectioning. } } If you have the necessary qualifications, please send your resume to : } Dr. H.N. Migeon } Director of LAM } 162a, Avenue de la Faiencerie } L-1511 Luxembourg } After a first selection, interviews will take place in Luxembourg. No } funding will be provided for overseas trips. } } Henri-Noel Migeon
Best regards,
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com
On Wed, 12 Mar 1997 WARRENJ1-at-cliffy.polaroid.com wrote:
} The 30 bits will give you additional shadow and highlight detail that } 24 bits will not provide. Now, you may ask 'my software can only } handle 24 bits so what good does 30 bits do when I will have to } convert it down?'. When you convert the file to 24 bits, the system } will choose from the best 24 bits to produce the converted file. I } would equate it to a survey or poll - the larger your sample, the more } accurate your results.
This statement seems a little bit tricky. 24 color bits gives one 256 grey shadows. If one want only to store and reproduce images - this is more than enough (128 is O'K for viewing). If one want to quantify grey levels for processing, 256 levels is also usually enough because of narrow linear range of film, grain, etc. Interpolation will give better results. If one want to waste money - I have no reasons not to this :)
Need to find a floor model lab vacuum coating systemwith : Defussion pump, HV(2000 Volt) 200 ma. feed thru, low voltage high current feed thru, cryotrap,etc. Also looking for an older Balzers 360 Freeze Fracture system(any condition) and an old hummer or a sputtering head for Au-Pd. By the way does anyone keep up with the "Particle-Protein" Freeze Fracture theories and can you send me some references? Been a while.
Jeff Day 3208 Statler Mesquite, Texas 75150 Day Phone:972-975-4338
Hello all, I have been looking at very thin (~10-30 A) multilayers of silicon oxide and nitride. I have very simplistically assumed that the position of the interface lies half-way between the first bright and dark fringe on each side of the interface. Does anyone know whether this is a valid assumption - and if there's any software that I can use to simulate the image? (Or, failing this, where I can get hold of the theory to let me do this myself).
Many thanks in advance,
Richard Beanland, GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK
Tina Carvalho wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have a question which has undoubtedly been answered before...! We are } going to buy a flat bed scanner, probably a Microtek with a transparency } adapter. We would primarily use it for scanning in TEM and SEM (polaroid) } negatives. Should we go for the 24-bit color, 300 x 600 dpi version, or } the more expensive (ouch) 30 bit color, 600 x 1200 dpi version? Our } resident computer guru says that we should go with the cheaper version } because we are unlikely to have an output source with better than 600 dpi } for most stuff. It's true; the printers around here are mostly 300 - 600 } dpi. Is there a compelling reason for the better model? Will I be really } sorry in a year if I don't? } } Thanks you all for all the expertise and advice on so many different } topics! } } Aloha, } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** Tina - buy the most true optical resolution and pixel depth you can afford - not so much for the SEM stuff but for the TEM negatives. The scanner should have enough optical (not interpolated) resolution to over sample the film resolution by at least a factor of 2.5 to 3 (the Nyquist sampling limit you know) otherwise you will not be able to treat the digitized images like the original negative - i.e. information will be lost! The company that makes tha scanner probably also makes a 1k x 2k version which would be even better.
} } Donald Lovett's explanation quoted below is very good except that it } perpetuates the myth that tonicity is related to the difference between } two OSMOLARITIES on opposite sides of the membrane. That is ONLY } true if the membrane involved is impermeable to ALL of the solutes } contributing to the osmolarity. ......
} Richard:
Thanks for your extended clarification of the subject. I agree entirely with your comments. I had tried to focus on the difference between the two terms and simplify my response with the caveat: } } "(a number of other factors affect this as well, but let's ignore them here)." } I also agree that the response of the cell to the solution (as evaluated by trial and error) is the most important aspect to microscopists, irrespective of how one names or measures the composition of the solution.
Don } } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } The College of New Jersey fax: (609) 771-2674 } Trenton, NJ 08650-4700 } } } } } }
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
I have found that what is 'adequate' for one opportunity may not be adequate for another. In any event, for those who are unsure of a decision between digital products based on varying specifications, I suggest you look at the end result. Take an image and scan it in on a 24 bit scanner and a 30 bit scanner, print them out on the printer(s) you would typically use and compare the results. If the 24 bit image is sufficient for your application, then buy the thing. The street price should be about $220. If you like the 30 bit converted to 24 better, then buy it. Its street price should be around $525.
As far as the comment regarding scanning 35 mm slides, I would not recommend using a flatbed scanner to scan 35mm images that will be enlarged more than 1x the original size, otherwise the image gets soft even with a 600 dpi scanner. If you are scanning 35mm on a regular basis, you should use a film scanner.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation
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On Wed, 12 Mar 1997 WARRENJ1-at-cliffy.polaroid.com wrote:
} The 30 bits will give you additional shadow and highlight detail that } 24 bits will not provide. Now, you may ask 'my software can only } handle 24 bits so what good does 30 bits do when I will have to } convert it down?'. When you convert the file to 24 bits, the system } will choose from the best 24 bits to produce the converted file. I } would equate it to a survey or poll - the larger your sample, the more } accurate your results.
This statement seems a little bit tricky. 24 color bits gives one 256 grey shadows. If one want only to store and reproduce images - this is more than enough (128 is O'K for viewing). If one want to quantify grey levels for processing, 256 levels is also usually enough because of narrow linear range of film, grain, etc. Interpolation will give better results. If one want to waste money - I have no reasons not to this :)
Hey, y'all Just wanted to thank everyone who replied to my request for recent job postings. Lots of people went digging through their trash and came up with the information that I needed. Thanks very much, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
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RE: David R. Stadden's inquiry (Where can I get one of those red antistatic guns, now that they are no longer handled by Fullam? Does anyone know of the English company that has this huge supply? Are there any distributors? Is there any similar product?)
I hope this is useful: Sigma Chemical Company (St. Louis, Missouri) carries the Zerostat 3 (approx $55; US catalog #Z10,881-2). Their UK contact info (from the list in the US catalog) is as follows:
In reguards to the question of tonicity, does anyone know the best percentage of sucrose to have in a primary Zamboni fixative(2% para and 15% picric acid in sorensons) to fix cell cultures in order to minimize any cell distortion.
Thanks in advance
Bob Underwood Morphology core University of Washington
In reguards to the question of tonicity, does anyone know the best percentage of sucrose to have in a primary Zamboni fixative(2% para and 15% picric acid in sorensons) to fix cell cultures in order to minimize any cell distortion.
Thanks in advance
Bob Underwood Morphology core University of Washington
} Our resident computer guru says that we should go with the cheaper } version because we are unlikely to have an output source with better } than 600 dpi for most stuff. It's true; the printers around here are } mostly 300 - 600 dpi. Is there a compelling reason for the better } model? Will I be really sorry in a year if I don't?
One reason to go with the more expensive one is that the 30bit depth will get you more contrast discrimination. Another reason is that, even with 300-600dpi printers, you might want to select a subregion of the scan and expand it to full page. In that case, the extra resolution of the better scanner will show.
At 05:04 PM 3/12/97 +1200, Richard Easingwood wrote: } Does anyone who performs vascular casting know where to obtain a product } called Mercox CL2? A fellow electron microscopist has heard that it is the } latest thing in vascular casting when mixed with methyl methacrylate but } hasn't been able to get any further info. } If anyone knows who makes it and where it can be bought we would like to hear.
There is a "Mercox Resin" in the Ladd catalog, but it doesn't say whether it's CL2.
Best regards, Steven Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Can formvar powder absorb water and cause holes in grid coatings, or is it just the solvent in which it is made that causes the problems? We have some rather old powder here and I wonder if it is still useable.
We recently purchased a film scanner to digitize images and diffraction patterns from 3.25"x4" negatives. Prior to this we had an old Microtek flatbed with a transparency adapter (300 dpi). It was ok as long as we didn't try to enlarge the image much.
I don't know how you will be using the digitized images. If all you want to do is digitize the negatives and then print the images without enlarging them, you don't need a high-resolution scanner. But I would recommend that you get a scanner with a larger bit-depth.
We often need to do some significant image analysis, so we needed a scanner which would give us very high optical resolutions and more than the usual 256 gray scale (24-bit color). We wanted at least 12 bits per color and 2400 dpi optical resolution. Our requirements may be more than you need.
I think John Warren's recommendation is good: try before you buy.
Russell E. Cook Scientific Associate 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
thanks to all who answered (either directly or via the listserver) my request for info on SIP refurbishment. I'll summarise these responses and make it available for anyone who contacts me directly. If there is enough interest I'll post the summary on the listserver. 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.
Always get the best you can afford. And, yes, you will be sorry if you don't! One of the uses I have found for the high resolution end of the scanner is to check small details (eg is that cell junction really tight?) in an EM negative without the hassle of a large print - it's amazing just how much it is possible to see on the computer in a fraction of the time. I have the ScanMaker 3 and am very happy with it. BUT, it is no good for 35mm, only prints and large format negatives.
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
} In reguards to the question of tonicity, does anyone know the best } percentage of sucrose to have in a primary Zamboni fixative(2% para and } 15% picric acid in sorensons) to fix cell cultures in order to minimize } any cell distortion.
I can't give a specific answer, but check out Maser MD et al: Relationships among pH, osmolality and concentration of fixative solutions, Stain Technology 42:175-182 (1967). You need to know what osmolality you want in the fixative solution. Around 300 milliosmols (range 200-400) seemed to be most common when I had to work it out too many years ago. In general the contribution of the fixative can be ignored. So, work out the osmolality of the buffer and add sucrose to bring the osmolality up to the total. For instance, Sorensen's is approx 105 at 0.05M, approx 210 at 0.1M. Sucrose is obvious (eg 0.2M = 200). It may be an idea to add some CaCl2 or MgCl2.
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
Best practice is to dry the powder at 60 deg.C for half an hour just before mixing it into the solvent. As for the solvent you ought to fractionate it first. Redistil it and monitor the temperature of the vapour at the stillhead and discard the (watery) fraction that comes off before the correct boiling point of the solvent is reached. Stop the still when there is around an (oily) tenth remaining of the original solvent. }
We supply the Zerostat, but in the US you could get one from Sigma. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au }
I know this question has been asked here before, so bear with me. Where } can I get one of those red antistatic guns, now that they are no longer } handled by Fullam? Their representative told me that there is supposedly } a hefty supply of these units still in the U.K., and that Fullam doesn't } handle them anymore because they'd have to buy more than they could sell. } I know Discwasher used to distribute them. Does anyone know of the } English company that has this huge supply? Are there any distributors? } Is there any similar product? Appreciate any leads. } } Dave Stadden } DRStad-at-Juno.com
Peggy Bisher wrote: ================================================= I am looking for 3.05mm rings that I can use as a TEM grid made of ceramic material. Does anybody know where I can find such thing? ================================================== About the closest thing about which I have any awareness would be our diamond grids and rings, 3 mm diameter. They are sized to fit into any TEM that takes a 3 mm grid. More information can be found about them on our website.
Disclaimer: These are products offered by SPI Supplies and we would stand to benefit if more people were using these diamond products.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
As several people have expressed an interest in a summary of responses to my SIP query I decided to post it to the listserver. My original posting was:
} I am about to replace the sputter ion pump elements in our two JEOL TEM's. } JEOL used to be able to recondition the elements used in their 2000FX } microscopes but no longer provide this service. Can anyone suggest another } company that does this sort of work? How long can I expect a reconditioned } unit to last compared with a new one and what sort of cost would be } reasonable given that JEOL are charging $2.5k and $4.5k (Australian } dollars) for new units? Thanks in advance for any advice you can give.
Several people recommended contacting the following company:
DUNIWAY STOCKROOM Corp. 800-446-8811 also 415-969-8811 fax 415-965-0764 1600 N. Shoreline Blvd. Mountain View CA 94043 also 1305 Space Park Way,Mountain View,CA 94043 info-at-duniway.com
Vacuum Scientific Services 44 Ellesmere St, manchester M15 4JY UK Tel. 44 161 833 9108 fax. 44 161 835 1443
Some of the comments received were:
A rebuilt pump is as good as a new one and should last as long. They open up the pump, clean it out, replace all the elements, and re-weld the case together. Prices are significantly less than buying new.
Duniway has a rebuilding service and also sell their own brand of pumps. They state that rebuilt pumps have the same performance as a new pump. If the pump has to be cut open to rebuild it, it can normally can be rebuilt 2 or 3 times. They also say that their pumps can normally be rebuilt 5-10 times. Being cut open is normally the case for small pumps. On large pumps the titanium elements can often be removed through the inlet port, allowing them to be rebuilt an unlimited number of times.
I emailed Duniway Stockroom Corporation and the following is an extract from the reply by Eroc Inman:
We do rebuild not only the elements for JEOL pumps, but JEOL actually sends all of their ion pumps to us for a complete overhaul/rebuild.
It your elements are just a standard size and configuration, like that of Varian ion pumps, then the pricing is quite easy to determine. The 150 l/s pump elements are generally US$390.00/each for rebuilding. This is the price for diode or triode configuration. I would be able to give you an absolute price once we had the opportunity to take a look at them. For now, I am quite certain that your elements are not any different than what we generally see.
I have not yet contacted the other companies. If anyone can supply an email address for them I would greatly appreciate it. I don't pretend that this is a comprehensive list of companies providing SIP refurbishment, nor do I recommend or have any affiliations with any one of them. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
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For the theory you need to look up Snell's Law and Molecular refractivity. The RI relates to the velocity of light through various substances. The difference between air and water RI results in a stick appearing angled at the interface. Transparent specimen become invisible if immersed in a liquid of identical RI. This is extensively used in police scientific work were glass fragments can be identified to come from a crime scene. Very simple, put a sample onto a microscope slide. Add a drop of RI solution. If the submersed specimen becomes invisible under the microscope, than the refractive index of the specimen is that of the RI test solutions. Some experimenting to find the right solution is required. Good luck. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au ---------------------------------------------------------------------------- ----- } From: Damian_Neuberger_at_RLT013-at-ccmailgw.mcgawpark.baxter.com } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Refractive Index } Date: Wednesday, 12 March 1997 6:49 } } I have just had a request to determine the refractive index of zinc } pyrithione, a crystalline, colorless, transparent powder. Does anyone } know what the RI is? It's not in McCrone Atlas. I will be getting a } sample Monday and with a set of Cargille RI liquids, I will determine } the RI.
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On 03/10/97 17:21:17 you wrote: } I would like tofind someone in the S.F. Bay area that has a freeze-fracture } machine that would be available for doing samples. I have a colleague in } the area who is looking for a facility where she might be able to do some } samples. } We would be very grateful for any leads. Thanks in advance. ML } } Mei Lie Wong } Department of Biochemistry } HHMI-UCSF } Ph. 415-476-4441 Fax 415-476-1902 } email wong-at-msg.ucsf.edu
Dear Mei, Please call Nancy Smith at Cal State Hayward (main number 510-885-3000). I am sure she cal help you. Regards
Igor C. Ivanov, SIMS,Auger,TEM,SEM Senior Scientist Analytical Services RJ LeeGroup, Inc. Contract Research 530 McCormick Str. (510)-567-0480 phone San Leandro, CA 94577 (510)-567-0488 FAX
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Thanks to all who responded to my cpd inquiry. The consensus seems to be that fresh, dry 100% ETOH is very important and that I need to keep my samples longer in the liquid CO2 to get a better exchange with the ETOH.
I'll try out all the suggestions. Hopefully, the only raisins I'll get from now on will be in my breakfast cereal. = )
Thanks again.
Paula = )
p.s. I'm going to phone some vendors of the resins & such to see if they've done studies as to which type of gloves work best. I'll keep you posted.
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
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Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) advertise a service for rebuilding sputter ion pumps (p. 25 of their catalog), and state that a properly rebuilt pump should "have the same performance as a new pump". The cost of rebuilding varies from about $400 to about $3000, depending on the size and type of pump No commercial interest - just trying to be helpful.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
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Dear Dr. Nunes:
Your question, "how to stain enzymes for EM" is so broad as to be unanswerable, I think. Which enzymes?, Plant tissue or animal? Cells grown in culture? I am sure someone on the list can help if your question is more specific.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu *************************************************************** ############################# Notice:
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Does anyone who performs vascular casting know where to obtain a product called Mercox CL2? A fellow electron microscopist has heard that it is the latest thing in vascular casting when mixed with methyl methacrylate but hasn't been able to get any further info. If anyone knows who makes it and where it can be bought we would like to hear.
Thanks in advance
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
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Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) advertise a service for rebuilding sputter ion pumps (p. 25 of their catalog), and state that a properly rebuilt pump should "have the same performance as a new pump". The cost of rebuilding varies from about $400 to about $3000, depending on the size and type of pump No commercial interest - just trying to be helpful.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
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In the theme of Diana van Driel's post check out Arborgh et al. "The osmotic effect of glutaraldehyde during fixation". J. Ultrastr. Res. 56:339-350, 1976. A very interesting and through study.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Thanks to all for your replies concerning measuring the refractive index of a crystalline powder using a refractometer. The replies indicated that it is not easy and has very specific requirement that I can not obtain. So I will use a set of standard refractive index liquids and a polarized light microscope which is the way I had originally planned to go.
Thanks again for the comments, you folks are a veritable deep well of knowledge.
Some time back, the Snappy framegrabber board, from Play, Inc. was recommended as an economical way to obtain digitized EM images. Well, acting on this recommendation, I purchased a Snappy board. However, I have been unable to obtain an acceptable stored image file, using the Snappy board (and it's associated software) in conjunction with a CCD camera and macro lens mounted above a light box on which the EM negative is mounted. Previously, this technique has worked well, when using another grabber board (costing around five times as much!) and NIH Image software. Has anyone obtained reasonable captured images of this type using the Snappy board? I fear that this application is beyond the resolution capabilities of the Snappy board, but there is always the possibility that I'm not setting something properly in the Snappy capture software.
Over the years I've found that uranyl acetate (dry powder/crystals) seem to degrade with time and find the staining greatly attenuated. It becomes less soluble and keeps for a shorter time in solution. Does anyone know the mechanism(s) of this change and how to perhaps prevent it? I do store dark, and have had it happen in both plastic and glass containers. I make my staining solutions up in small volumes. I've ended up having more picked up by EH&S than I've ever really used. If it's inevitable, maybe we could ask our vendors to offer smaller quantities as an alternative......Grace Kennedy
Please do not reply via eMail but direct your resume to: M R Jobe Salaried Personnel THE GOODYEAR TIRE & RUBBER COMPANY 1144 E. Market Street Akron, Ohio 44316
The Goodyear Tire & Rubber Company, a world leader in tire development and manufacturing, has an excellent opportunity for a professional skilled in Microscopy in its Analytical Sciences Department, Corporate Research Division, located in Akron, Ohio. This position involves the analysis and characterization of polymers and materials used in the tire and rubber industry.
Qualified candidates must have a PhD or MS with 3 years experience in light, electron and atomic force microscopy with a strong background in image analysis. Current experience should include techniques employed in polarized light, brightfield, phase contrast, fluorescence, interference microscopy and photomicrography. Knowledge of transmission and scanning electron microscopy and EDS systems is also required. Familiarity with failure analysis and mixing studies of rubber, plastics and composites is desirable.
This position with a Fortune 100 industry leader offers excellent benefits, relocation assistance and competitive salary commensurate with education and experience. If you have the qualifications and desire to meet the rewarding challenges presented by Goodyear, direct your resume to:
M R Jobe Salaried Personnel THE GOODYEAR TIRE & RUBBER COMPANY 1144 E. Market Street Akron, Ohio 44316
An Equal Opportunity Employer, M/F/D/V Applicants must be lawfully authorized to work in the U.S.
L E Porter Phone (330) 796-1620 Head of Microscopy Fax (330) 796-3304 The Goodyear Tire & Rubber Company EMail LPORTER-at-GOODYEAR.COM Dept 415A 142 Goodyear Blvd Akron, OH 44305 USA
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Good Afternoon (Central Std Time USA) all Microscopists:
I have just had a request to determine the refractive index of zinc pyrithione, a crystalline, colorless, transparent powder. Does anyone know what the RI is? It's not in McCrone Atlas. I will be getting a sample Monday and with a set of Cargille RI liquids, I will determine the RI.
Alternatively, I have a B & L Abbe type refractometer used to determine refractive index of liquids. However, I thought I read somewhere that it could be used to determine the RI of solids. If this is in fact possible, does anyone out there know how?
My wife is having to do a poll of scientist for one of her certification classes. She has taught science in middle and high school. For the philosophers in the crowd I hope that you can have fun with this one, if you could help out with her survey it would be much appreciated. For all others, I ask your indulgence. Please forward you replys to me and I will deliver them. The survey follows.
Thank you, Chuck
SURVEY QUESTIONS
Name: Degree/Profession:
1. How would you describe the Nature of Science?
2. Describe the ideal classroom and curriculum for scientific learning.
3. What role do you believe the History of Science should play in teaching science in the classroom?
END SURVEY ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-7996 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
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I am acquiring an ISI SX-30E and need a copy of the manual for this instrument without paying the hundreds of dollars Topcon wants for such a manual. Any suggestions where I might be able to get a xerox of the manual or borrow a copy to xerox. Any help would be appreciated.
Jim Ekstrom Phillips Exeter Academy
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Hello Fellow Listserv Members, For the past 15 or so years I have been using a wonderful film by Kodak which makes very nice black and white slides of continous tone and high contrast copy. This is a positive film which requires no reversal processing. It is called Kodak Direct Duplicating Microfilm 2468. I have been buying it from a supplier in Florida who has apparently gone out of business. Now here is the catch. Kodak, according to my long time photography needs supplier, is not willing to sell this film in anything less than a case. There are 50 100ft rolls in a case at about $40.00 per 100ft roll. My question is....are there any of you out there who might be familiar with this film and know of suppliers other than Brandon's in Jacksonville, FLA.? Brandon's is the supplier from whom I have ordered this film in the past. Or another question which has just come to mind...are there any EM Suppliers out there who might be interested in this film as one of their catalogue items? I would appreciate hearing from any of you who might have information on where to find this film or someone who might be interested in purchasing a case for distribution. I trust this is an appropriate post. Since we all make slides from time to time, I felt that it might be. Perhaps it would be better to reply to me directly and if there are others interested, let me know and I will pass on any information I receive. Thank you very much, Sandra Zane Sandra F. Zane, EM Tech. sfzane-at-email.uncc.edu Dept. of Biology, UNCC Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
We just received a Polaroid Sprintscan 45 for digitizing negatives from our microscopes. Preliminary trials look good. Unfortunately, no negative holder is available for TEM negatives (3 1/4 x 4 inch). This is really unfortunate since we have hundreds of such negs to scan and using the 4x5 holder isn't very user friendly - although it can be done. Has anyone had experience with this system? Any workarounds?
I suggested to Polaroid that if they would provide me with a couple of their standard 2 1/4 x 2 3/4 inch adapters, that I could have our shop mill out the proper size frame. I was shocked to find that not only were no plans in place to make the TEM holders but that Polaroid would not provide even the stock holders ( or even the metal) for modification. Caveat emptor.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
About 6 months ago I asked the group a similar question about the Snappy and got a number of good responses concerning quality of cable, sources of noise, etc.
I have sent a rather long summary of all these replies to the originator of this post, and just wanted to make everyone else aware it is available. Just E-mail me and I'll forward it on to you individually (didn't want to bother the whole listserve with such a long message).
-Karen
On March 14, 1997, demczyk-at-erxindy.rl.plh.af.mil wrote:
} Some time back, the Snappy framegrabber board, from Play, Inc. } was recommended as an economical way to obtain digitized EM images. } Well, acting on this recommendation, I purchased a Snappy board. However, } I have been unable to obtain an acceptable stored image file, using } the Snappy board (and it's associated software) in conjunction } with a CCD camera and macro lens mounted above a light box on which the } EM negative is mounted. Previously, this technique has worked well, } when using another grabber board (costing around five times as much!) } and NIH Image software. Has anyone obtained reasonable captured images of } this type using the Snappy board? I fear that this application is beyond } the resolution capabilities of the Snappy board, but there is always the } possibility that I'm not setting something properly in the Snappy capture } software. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Life Sciences Sector Lab Reply: kszaruba-at-mmm.com 3M Company 3M Center 270-1S-01 Phone: 612-737-2971 St. Paul, MN 55144-1000 Fax: 736-1519
These opinions are my own and may not represent those of 3M.
I am sorry that you were given the information that you were given. I recommended earlier this year that we offer a carrier for TEM negatives due to the lack of other currently available options. The last word I heard was that we are working on a carrier that will be adjustable for various film sizes. They also are working on creating a carrier dedicated to the TEM format you mentioned. As a temporary solution, cutting a mask that would fit in the 4x5 holder with the proper opening should work.
I am going to forward your message to the Worldwide Product & Technical Managers for Polaroid Scanners.
I'll keep you posted.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation
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We just received a Polaroid Sprintscan 45 for digitizing negatives from our microscopes. Preliminary trials look good. Unfortunately, no negative holder is available for TEM negatives (3 1/4 x 4 inch). This is really unfortunate since we have hundreds of such negs to scan and using the 4x5 holder isn't very user friendly - although it can be done. Has anyone had experience with this system? Any workarounds?
I suggested to Polaroid that if they would provide me with a couple of their standard 2 1/4 x 2 3/4 inch adapters, that I could have our shop mill out the proper size frame. I was shocked to find that not only were no plans in place to make the TEM holders but that Polaroid would not provide even the stock holders ( or even the metal) for modification. Caveat emptor.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Have you tried Polaroid's 35mm Polagraph Instant Slide Film? It is a high contrast b&w film that you can process & mount at your desk. The processing does require our processor - either power or manual. The film comes with the development chemistry. You just have to buy the reusable slide mounts separately. It is a little denser than wet processed film, but it may not be as apparent with a high contrast image.
John Warren Area Sales Manager Digital Products Polaroid Corporation
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Hello Fellow Listserv Members, For the past 15 or so years I have been using a wonderful film by Kodak which makes very nice black and white slides of continous tone and high contrast copy. This is a positive film which requires no reversal processing. It is called Kodak Direct Duplicating Microfilm 2468. I have been buying it from a supplier in Florida who has apparently gone out of business. Now here is the catch. Kodak, according to my long time photography needs supplier, is not willing to sell this film in anything less than a case. There are 50 100ft rolls in a case at about $40.00 per 100ft roll. My question is....are there any of you out there who might be familiar with this film and know of suppliers other than Brandon's in Jacksonville, FLA.? Brandon's is the supplier from whom I have ordered this film in the past. Or another question which has just come to mind...are there any EM Suppliers out there who might be interested in this film as one of their catalogue items? I would appreciate hearing from any of you who might have information on where to find this film or someone who might be interested in purchasing a case for distribution. I trust this is an appropriate post. Since we all make slides from time to time, I felt that it might be. Perhaps it would be better to reply to me directly and if there are others interested, let me know and I will pass on any information I receive. Thank you very much, Sandra Zane Sandra F. Zane, EM Tech. sfzane-at-email.uncc.edu Dept. of Biology, UNCC Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
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Does anyone who performs vascular casting know where to obtain a product called Mercox CL2? A fellow electron microscopist has heard that it is the latest thing in vascular casting when mixed with methyl methacrylate but hasn't been able to get any further info. If anyone knows who makes it and where it can be bought we would like to hear.
Thanks in advance
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
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Thanks to all who responded to my cpd inquiry. The consensus seems to be that fresh, dry 100% ETOH is very important and that I need to keep my samples longer in the liquid CO2 to get a better exchange with the ETOH.
I'll try out all the suggestions. Hopefully, the only raisins I'll get from now on will be in my breakfast cereal. = )
Thanks again.
Paula = )
p.s. I'm going to phone some vendors of the resins & such to see if they've done studies as to which type of gloves work best. I'll keep you posted.
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
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This message was found in a dead-letter box and appears to be for you. If you have already gotten a copy of this message, we beg your tolerance.
Does anyone have any experience with this microscope. Is this considered a fine microscope for the microscopy hobby? Is this scope lacking any of the essentials or necessities for the amateur microscopist? I was surfing the net under microscopes and this one really caught my attention. Any comments would be greatly appreciated.
A few days ago someone asked about having sputter ion pumps rebuilt. It occurs to me to comment that sometimes it is possible to squeeze a few months of extra use out of a pump before rebuilding becomes absolutely necessary. Frequently, pumps become unstable because a low impedence path develops between the anode and cathode structures, usually because a flake of Ti breaks off the anode and lodges between it and the cathode, or because of the growth of a Ti whisker between the two electrodes. Sometimes such a condition can be relieved temporairly by turning the power to the pump on for a few seconds (ONLY a FEW) when the pressure in the pump is just above 1 Pa (0.01 Torr), repeating the process three or four times, if necessary. (See Vac. Methods in EM, p 295) If this treatment is successful, the pump's performance will stabalize, and the pump will perform satisfactorily again, but only for a limited time - ultimately, of course, it will need to be reconditioned.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Hi, Help! I just started to use the NIH-image to analyze the distribution of gold particles on the EM negatives. The software was downloaded from the web site and it is NIH 1.61. My problem at this point is that I can not open my scanned image to the proper size I wanted. I tried to acquire, open or import the image, the result is the same -- a very small picture show up on the screen. When this image was magnified, the pixels came out which make particle counting impossible. Apparently, it is not the problem with the scanner since the image could be processed nicely with Adobe. But if I open the Adobe processed image through NIH, the same thing happens as described above. I also tried to scan the negatives using higher ppi, approximately 1000 ppi to incease the resolution. That did not help. Any suggestions and help will be greatly appreciated.
When I label LR White thin sections of decalcified bone with immunogold I get a fine contamination - dirt - that is not present on my conventional EM/epon sections. The contaminant is a fine floccular stuff made up of filament- like strands, each less than 10 nm in diameter, that clump togethr into irregular shapes of widely ranging size. The contaminant is not seen on the phosphorescent viewing screen but definitely shows up in the negative and print. All diluents and wash solutions are filtered with 0.2 micron Gelman Acrodisc syringe filters. Antibodies are diluted either in Tris- saline or BioMeda Primary Antibody Diluent; blockers are serum, fish gelatin, BSA. Wash solutons ar tris-saline. All solutions have Triton X. Final staining is with methanolic or aqueous uranyl acetate. Deleting uranyl acetate staining still leaves contamination. Deleting primary antibody still leaves contamination. LR White alone without processing for immuno and stained with uranyl acetate is clean. So, something in the gold, or in the secondary antibody may be doing it, but I haven't checked this out yet. Any ideas on what this dirt could be and how to get rid of it would be appreciated. Sorry for the long-windedness...and Thanks.
Pat Masarachia Bone Biology Merck Research Labs West Point, PA tel:215-652-7999 e-mail: pat_masarachia-at-merck.com
The Central Microscopy Research Facility at the University of Iowa will host an immunocytochemistry workshop given by Jan Leunissen and Peter Van der Blass. It is to be held May 9-10, 1997 in Iowa City. For more information, visit our website at: http://www.uiowa.edu/~cemrf/cemrf/jan_workshop.html or contact Kenneth Moore (319)-335-8143, kenneth-moore-at-uiowa.edu
Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
NIH Image is extensively used in the Integrated Microscopy Resource (IMR). We never have this kind of problem. If you want, I can FTP you some of my images. In this way you can easily find where the problem is. Hope that will help you.
Ya Chen
Ya Chen
========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
Does anyone have available (new or used) a double tilt holder, a double tilt heating holder, a double tilt cooling holder and a selective aperture set for the LEO/Zeiss 910 TEM? If so, please let me know (incl. price).
Sandra Zane was wondering where she could get Kodak Direct Duplicating Microfilm 2468. We use Eastman 5360 which sounds like the same thing; 35 mm, gives a positive without a complicated kit, developes in Dektol. We order it locally (I think) or you can get it from Freestyle Sales Co. 5124 Sunset Blvd. Los Angeles, CA 90027, www.freestylesalesco.com. In their catalogue it is called "Kodak B&W duplicating film 5360" and is available in 50 or 100 foot rolls or 4x5 sheets.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
POSTDOCTORAL POSITION is available immediately to develop three dimensional image analysis algorithms for quantitative analysis of cells in tumor specimens. Experience in image analysis and UNIX workstations, and a strong math background are essential. Candidate should have a recent Ph.D. in the physical sciences, or Computer Science and should be an expert in developing code in C/C++. Send curriculum vitae and the names of three referees to:
Dr. Ravi Malladi, MS 50A-2152 Lawrence Berkeley National Laboratory University of California, Berkeley, CA 94720.
Forgive my inexperiece and knowledge on this new hobby of mine but can someone briefly explain the differece between semi-plan objectives and plan objectives. Under what applications would one type be more advantageous over the other type. Thanks for any information you may have on this subject.
Hope I'm not duplicating an earlier topic but couldn't find anything in postings kept over the last few months so, here goes...
What factor (s) in an immungold labeling protocol has (have) the greatest impact on background, e.g.. antibody dilution, buffer, etc.? I'm looking at the immunogold protocol itself so I can use valuable tissue already embedded.
(I'm labeling virus-infected plant leaf tissue with monoclonal antibodies to viral proteins - samples are embedded in LR White and fixed in 3% paraformaldehyde/0.2% glutaraldehyde. We block before primary antibody incubation with a TRIS-HCl buffer containing bovine serum albumin and goat normal serum. Our secondary antibody is a commercially prepared goat anti-mouse gold conjugate. Usually we have excessive background on chloroplasts, mitochondria and cell walls; if we get rid of background by increasing dilutions of antibodies, we lose specific reactions as well).
One interesting point....we don't have this problem with polyclonal antibodies, only monoclonals.
Thanks for any tips -- this is really frustrating!
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
} Date: Mon, 17 Mar 1997 16:33:06 -0800 } From: Geoff McAuliffe {mcauliff-at-UMDNJ.EDU} } To: MSA Listserver {Microscopy-at-Sparc5.Microscopy.Com} } Subject: re: positive film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear List: } } Sandra Zane was wondering where she could get Kodak Direct } Duplicating Microfilm 2468. We use Eastman 5360 which sounds like the 0} same thing; 35 mm, gives a positive without a complicated kit, developes } in Dektol. We order it locally (I think) or you can get it from Freestyle } Sales Co. 5124 Sunset Blvd. Los Angeles, CA 90027, } www.freestylesalesco.com. In their catalogue it is called "Kodak B&W } duplicating film 5360" and is available in 50 or 100 foot rolls or 4x5 } sheets. } } Geoff } -- } *************************************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane Piscataway, NJ 08854 } voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu } *************************************************************** } We have good success with 5360 also, and it's easy to get. Here is a copy of information I sent directly to Sandra:
The Direct MP Film 5360 is cat # 24611 from Ted Pella, $22.90/100 ft roll. 800 237-3526 or tedpel-at-aql.com or Fax 916 243-3761 I put the camera on B and manually work the shutter for 2, 2.5, 3 sec, depending on the size (height of camera). It's so cheap, I usually bracket and shoot 3 shots for each to make sure I don't have to repeat shooting. Most of the time, any of the shots could be used, but I pick the ones that are most closely matched for use in a single presentation. I develop in D-19 4 min at 20 oC. There is another film, Kodak Contrast Copy Film, that works about the same, but gives a bluer tint. I only keep the 5360 now.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I have done a lot of LR White immunostaining and have had many kinds of dirt problems. It sounds like you are doing nearly everything right but you didn't mention doing a final rinse in filtered distilled water after your gold secondary. I rinse 2-3x in my buffer after secondary then do a fairly vigorous rinse in the beaker of distilled filtered water, then dry the grids, then go to staining.
You may also check your water and photoflo source and see if it is actually on your negatives.
Bob Morphology Core U.of Washington Seattle
On Mon, 17 Mar 1997, Pat Masarachia wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } When I label LR White thin sections of decalcified bone with } immunogold I get a fine contamination - dirt - that is not present on } my conventional EM/epon sections. } The contaminant is a fine floccular stuff made up of filament- } like strands, each less than 10 nm in diameter, that clump togethr } into irregular shapes of widely ranging size. The contaminant is not } seen on the phosphorescent viewing screen but definitely shows up } in the negative and print. } All diluents and wash solutions are filtered with 0.2 micron } Gelman Acrodisc syringe filters. Antibodies are diluted either in Tris- } saline or BioMeda Primary Antibody Diluent; blockers are serum, fish } gelatin, BSA. Wash solutons ar tris-saline. All solutions have } Triton X. Final staining is with methanolic or aqueous uranyl acetate. } Deleting uranyl acetate staining still leaves contamination. } Deleting primary antibody still leaves contamination. LR White alone } without processing for immuno and stained with uranyl acetate is clean. } So, something in the gold, or in the secondary antibody may be } doing it, but I haven't checked this out yet. Any ideas on what this } dirt could be and how to get rid of it would be appreciated. } Sorry for the long-windedness...and Thanks. } } Pat Masarachia } Bone Biology } Merck Research Labs } West Point, PA } tel:215-652-7999 } e-mail: pat_masarachia-at-merck.com } } }
I would think if your minus primary control are clean then your primary antibody is non-specificly binding to something in the plant tissue. Usually the primary has the most effect on backround. Is your antisera purified?
Bob Morphology Core U of Washington Seattle
On Tue, 18 Mar 1997, Peggy Brannigan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello immunogold experts! } } Hope I'm not duplicating an earlier topic but couldn't find anything in } postings kept over the last few months so, here goes... } } What factor (s) in an immungold labeling protocol has (have) the greatest } impact on background, e.g.. antibody dilution, buffer, etc.? I'm } looking at the immunogold protocol itself so I can use valuable tissue } already embedded. } } (I'm labeling virus-infected plant leaf tissue with monoclonal antibodies } to viral proteins - samples are embedded in LR White and fixed in 3% } paraformaldehyde/0.2% glutaraldehyde. We block before primary antibody } incubation with a TRIS-HCl buffer containing bovine serum albumin and goat } normal serum. Our secondary antibody is a commercially prepared goat } anti-mouse gold conjugate. Usually we have excessive background on } chloroplasts, mitochondria and cell walls; if we get rid of background by } increasing dilutions of antibodies, we lose specific reactions as well). } } } One interesting point....we don't have this problem with polyclonal } antibodies, only monoclonals. } } Thanks for any tips -- this is really frustrating! } } } Peggy } } Peggy Brannigan } Electron Microscopy } Floral and Nursery Plants Research Unit } National Arboretum } } Bldg. 010A R.238 } 10300 Baltimore Avenue } Beltsville, MD USA20705 } } Phone: (301) 504-6097 } Fax : (301) 504-5096 } Email: brannign-at-asrr.arsusda. gov } } } }
We also find that immuno-EM is problematic with MAbs. Are you using ascites or an IgG prep? I would use IgG if possible (the EZ Sep products from Pharmacia Biotech are handy for isolating IgG from ascites, MHO). You may want to try adding 0.05% Tween-20 to your blocking and antibody soln's. This often helps bring down the background. The downside (there's always a downside) is that the tween also tends to wash a lot of the contrast out of the sections -- at least it does for our LRGold embedded material. You may need to adjust the length of time you let the grids float on the antibody sol'n to give good label without too much loss of contrast. 1-4 hours on the primary is a good range for starters.
Hope this helps,
Greg Martin Dept. of Cell Biology and Anatomy Johns Hopkins School of Medicine
On Tue, 18 Mar 1997, Peggy Brannigan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello immunogold experts! } } Hope I'm not duplicating an earlier topic but couldn't find anything in } postings kept over the last few months so, here goes... } } What factor (s) in an immungold labeling protocol has (have) the greatest } impact on background, e.g.. antibody dilution, buffer, etc.? I'm } looking at the immunogold protocol itself so I can use valuable tissue } already embedded. } } (I'm labeling virus-infected plant leaf tissue with monoclonal antibodies } to viral proteins - samples are embedded in LR White and fixed in 3% } paraformaldehyde/0.2% glutaraldehyde. We block before primary antibody } incubation with a TRIS-HCl buffer containing bovine serum albumin and goat } normal serum. Our secondary antibody is a commercially prepared goat } anti-mouse gold conjugate. Usually we have excessive background on } chloroplasts, mitochondria and cell walls; if we get rid of background by } increasing dilutions of antibodies, we lose specific reactions as well). } } } One interesting point....we don't have this problem with polyclonal } antibodies, only monoclonals. } } Thanks for any tips -- this is really frustrating! } } } Peggy } } Peggy Brannigan } Electron Microscopy } Floral and Nursery Plants Research Unit } National Arboretum } } Bldg. 010A R.238 } 10300 Baltimore Avenue } Beltsville, MD USA20705 } } Phone: (301) 504-6097 } Fax : (301) 504-5096 } Email: brannign-at-asrr.arsusda. gov } } } }
I am looking for a person to work on an exciting new project.
The appointment could be at the post-doctoral level or at other levels according to the background of the person appointed. In any case the post will be for three years.
The job is at the Materials Research Laboratory of the University of Illinois at Urbana.
The project is a joint enterprise involving Argonne National Lab, Oak Ridge National Lab, the Lawrence Berkeley Lab and NIST as well as the University of Illinois. The Project has the aim of developing a new kind of environment for electron microscopy and related techniques, in which the instruments can be operated remotely with the same effectiveness as they can be operated in the instrument room. More details of the project can be found at http://tpm.amc.anl.gov/MMC MMC is the abbreviation of the project name.
I am looking for someone who has familiarity with electron microscopy (preferably TEM) or a closely related technique - and who has well developed interests and experience in computing, particularly the interfacing of instruments for computer control and/or the networking of images.
Will any one interested please contact me right away. We would like the job to be started as soon as possible. ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones) **
You are invited to attend the 1997 Annual Spring meeting of the Arizona Imaging and Microanalysis Society. The meeting will be Thursday March 27th and Friday March 28, 1997. Meetings will be held in the Senior Ballroom of the University of Arizona's Student Union Building. Admission is free to interested participants.
Please pass this information on to interested colleagues.
========================================================================= DIGITAL IMAGING =========================================================================
Digital imaging has found its way into many scientific fields (biology, engineering, astronomy, radiology, etc.). As a wider range of researchers begin to use these techniques, they are often unsure of where to begin and are unaware of the pitfalls involved in the field of digital imaging. This workshop will introduce you to some important concepts and issues relevant to digital imaging.
Thursday, March 27, 1997
9:00 Welcome and overview of the meeting Doug Cromey, President, AIMS
9:15 Image acquisition: General approaches, common problems and issues relating to CCD cameras. Scott Sternberg, Photometrics
10:15 Coffee break
10:30 Data output devices John Spenseri, Micrographix West
11:15 Data storage, movement and archiving Doug Cromey, President, AIMS
6:00 Buffet dinner in the Student Union's Sr. Ballroom (Mexican-style entrees, Preregister with Patty Jansma, cost $11.00)
Friday, March 28, 1997
9:00 Ethics and image analysis: Panel discussion. (Questions from the audience are welcome.)
Panel members: John Gilkey, UA, Pharmacology Marvin Landis, UA, CCIT- User Support David Ring, Dir. Photography, Biomedical Communications, UA Mary Rykowski, Assist. Prof., Cell Biology and Anatomy, UA Scott Sternberg, Photometrics
10:30 Coffee Break
10:45 Student platform presentations
11:45 Lunch. AIMS Business Meeting New officers installation.
1:00 Student platform presentations
3:00 Presentation of student awards and closing remarks
A histochemistry query: In the course of our work, we examine many different vertebrate tissues (brain, skin, liver, kidney, etc) embedded in plastic resin (Eponate or LR White). We currently do a lot of LM viewing of 2 micron sections, using toluidine blue as a counterstain. Some tissues have been fixed with aldehydes and osmium, for subsequent EM thin section studies. For immunocytochemistry, other tissues are only lightly fixed with aldehydes.
Our question: Can you suggest other useful counterstains, besides toluidine blue, that are compatible with plastic sections. If you have a favorite recipe, please send to us off-line, so that we don't clog the listserver. We'll compile a list of the answers we receive, for those who are interested.
Thanks in advance
David H. Hall hall-at-aecom.yu.edu Dept of Neuroscience Albert Einstein Col Medicine, Bronx, NY 10461 (718) 430-2195 (718) 430-8821 FAX
A formal proposal to create a new group tentatively called sci.bio.immunocytochem has just been posted to news.announce.newgroups. This is a reposting of that proposal.
REQUEST FOR DISCUSSION (RFD) unmoderated group sci.bio.immunocytochem
This is the 2nd Request For Discussion (RFD) for the creation of a world-wide unmoderated Usenet newsgroup sci.bio.immunocytochem, currently being discussed in news.groups
Suggestions for improvements to this proposal are welcome. Discussion about it should take place in news.groups. A vote is expected to be held in about three to four weeks.
This is not a Call for Votes (CFV); you cannot vote at this time. Procedural details are below.
CHANGES from previous RFD:
2nd RFD posted because more than 60 days have elapsed since 1st RFD. There are (minor changes in Distribution and Newsgroup line.
Newsgroup line: sci.bio.immunocytochem Immuno-labelling of biological material.
RATIONALE: sci.bio.immunocytochem
Immunohistochemists and immunocytochemists already enjoy the benefits of online communication, utilizing e-mail, accessing web sites, and subscribing to specialised mailing lists. Usenet newsgroups are also popular, but this is less obvious because articles with immunocytochemical/immunohistochemical content get posted to many different newsgroups. Most articles are posted to a favourite five or six newsgroups including bionet.cellbiol, sci.med.immunology and sci.techniques.microscopy, but often articles get posted to any one of fourteen or fifteen newsgroups in the sci. and bionet. heirarchies. Some of these are listed in the distribution list at the end of this proposal.
In my view, no existing newsgroup fulfils the criteria necessary to attract all the various immunocytochemistry postings. I do not wish to draw users away from other newsgroups, only to encourage scientists to share their knowledge and expertise on immunocytochemistry in the most effective manner. In response to my proposal to create a newsgroup dedicated to the discussion of immunocytochemistry and immunohistochemistry, I have received e-mail and faxes from researchers all over the world offering their support and encouragment.
Immunocytochemistry and immunohistochemistry are not subdivisions of immunology, molecular biology or chemistry. Microscopy, although essential, is only a small part of the story. Immunocytochemistry and immunohistochemistry are multi- disciplinary, therefore discussions are destined to stay distributed amongst the different newsgroups until they are all brought together under one umbrella. This would then act as a focus point for all the immunocytochemists who are already Internet users, and encourage new subscribers to Usenet.
CHARTER: sci.bio.immunocytochem
This is a newsgroup for the exchange of information relating to immunocytochemistry and immunohistochemistry. This unique research tool is used to locate and identify specific molecules in biological material, at the microscopical level.
Articles posted to this group must be relevant to one or more aspects of the above. The kind of subjects that may be discussed include techniques, theory, presentation of results, requests for collaboration, history, equipment, publication references, notice of events, tips and trouble-shooting, jobs offered andwanted, jokes, stories and new ideas, so long as the posting bears a direct relevance to the central theme. There will be a list of Frequently Asked Questions (FAQs) to help newcomers.
A relevant posting could just be a simple question or answer, for example "Has anyone got any experience with this reagent ?"or "Which course could I attend to learn more about immunogold labelling?". There will be articles reminding people to read the list of FAQs prior to posting their own article. Usenet readers may get involved in complex discussions about, for example, multiple labelling, proper use of control experiments, microwave antigen retrieval or quantitative measurements. Remember that articles posted to a newsgroup are intended for a wide readership, so if you have information which concerns only one or two people then please don't use this newsgroup, use e-mail.
Commercial advertisements for services, equipment or reagents violate the charter unless one or more of the following apply: (a)The advertisement is part of a comprehensive article designed specifically to address issues raised in earlier articles posted to the group (b)A general reference to the type of product does not suffice for technical reasons and it is necessary to specify the exact commercial product (c) The information is offered primarily for the benefit of the readers (d)The advertisement is for second-hand equipment specific to immunocytochemistry (e) Requests or offers for free products are acceptable if they are not part of a sales promotion.
END CHARTER.
PROCEDURE:
This is a request for discussion, not a call for votes. In this phase of the process, any potential problems with the proposed newsgroups should be raised and resolved. The discussion period will continue for a minimum of 21 days (starting from when the 2nd RFD for this proposal is posted to news.announce.newgroups), after which a Call For Votes (CFV) may be posted by a neutral vote taker if the discussion warrants it. Please do not attempt to vote until this happens.
All discussion of this proposal should be posted to news.groups. This RFD attempts to comply fully with the Usenet newsgroup creation guidelines outlined in "How to Create a New Usenet Newsgroup" and "How to Format and Submit a New Group Proposal". Please refer to these documents (available in news.announce.newgroups) if you have any questions about the process.
DISTRIBUTION:
This RFD has been posted to the following newsgroups: news.announce.newgroups,news.groups,bionet.cellbiol, bionet.diagnostics,bionet.immunology,bionet.microbiology, bionet.molbio.methds-reagnts,sci.bio.misc,sci.med.immunology, sci.techniques.microscopy
This RFD will be reposted to the following newsgroups after its posting in news.announce.newgroups: bionet.molbio.proteins,bionet.neuroscience,bionet.plants, sci.bio.microbiology,sci.med,sci.med.laboratory,sci.misc, sci.nanotech
This RFD will also be reposted to the following mailing lists after its posting in news.announce.newgroups:
Histonet mailing list: {histonet-at-pathology.swmed.edu} Information pertaining to the technical aspects of histology and histopathology such as tissue fixatives and processing, routine histology, special stains, immunohistochemistry, in-situ hybridization etc. To subscribe type "subscribe digest" into the subject box and leave the text box empty, or to subscribe to the full service just type "subscribe". For more info access web site http://www.mwrn.com/subject/histonet.htms
Microscopy Society of America listserver: Questions/comments/answers in the various fields of Microscopy Currently over 3000 subscribers. To subscribe send the message "subscribe" to {Listserver-at-MSA.Microscopy.Com} then send messages in plain text to {Microscopy-at-MSA.Microscopy.Com} For more info access web site http://www.amc.anl.gov/ Docs/anl/Nestor/Software/telecommList.html
Stanford University list server To subscribe, send a message to {majordomo-at-pathology.stanford.edu} with "subscribe ipox-l" in the body of your message. This list helps pathologists and other laboratory professionals to exchange information about immunoperoxidase methods.
This RFD will also be reposted to the following web-sites after its posting in news.announce.newgroups:
Royal Microscope Society http://www.rms.org.uk Web Master Dr R. A. D. Mackenzie {r.a.mackenzie-at-open.ac.uk}
Center for Cell Imaging Department of Cell Biology Yale University School of Medicine Introduction to Immunocytochemistry http://info.med.yale.edu/cellimg/CCIimmuno.html Web Master Paul Webster { paul_webster-at-yale.edu}
Proponent: Amanda Wilson {awilson-at-aw.u-net.com} Proponent: Paul Monaghan { monaghan-at-icr.ac.uk} Mentor: Jonathan Grobe {grobe-at-netins.net}
Amanda Wilson Deputy Manager, E.M. Unit St George's Hospital Medical School, S.W.London, UK Tel: 0181 725 5220 (work) e-mail {awilson-at-aw.u-net.com}
I am in need of an X-unit (EHT regulator) for our Philips Model 300. Does anyone happen to know where I might find a working unit? Please respond directly to me. Thanks in advance!
Scott Schwinge University of Washington Friday Harbor Labs 360-378-2165 schwinge-at-fhl.washington.edu
In our experience the following are the most important issues for good antibody staining-
quality of primary ab } fixative } buffer } dilution } blocking agents.
Most of our background problems can be corrected by getting a better antibody. Some out there work great on gels but once in among all the cellular proteins they start binding to everything.
The proper fixation (which of course varies for every different antibody antigen mix we try) is the next most critical.
We have also found differing the buffers can increase the signal to noise ratio. Although we normally use Gey's salts (it seems to works well with a whole range of antigens) sometimes cacodylate is superior.
The bottom line, however, is that with any new antigen or antibody it takes us a lot of trial and error to find the correct match.
Although we have not yet discovered a magic potion, I hope this limited answer is helpful-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Tue, 18 Mar 1997, Peggy Brannigan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello immunogold experts! } } Hope I'm not duplicating an earlier topic but couldn't find anything in } postings kept over the last few months so, here goes... } } What factor (s) in an immungold labeling protocol has (have) the greatest } impact on background, e.g.. antibody dilution, buffer, etc.? I'm } looking at the immunogold protocol itself so I can use valuable tissue } already embedded. } } (I'm labeling virus-infected plant leaf tissue with monoclonal antibodies } to viral proteins - samples are embedded in LR White and fixed in 3% } paraformaldehyde/0.2% glutaraldehyde. We block before primary antibody } incubation with a TRIS-HCl buffer containing bovine serum albumin and goat } normal serum. Our secondary antibody is a commercially prepared goat } anti-mouse gold conjugate. Usually we have excessive background on } chloroplasts, mitochondria and cell walls; if we get rid of background by } increasing dilutions of antibodies, we lose specific reactions as well). } } } One interesting point....we don't have this problem with polyclonal } antibodies, only monoclonals. } } Thanks for any tips -- this is really frustrating! } } } Peggy } } Peggy Brannigan } Electron Microscopy } Floral and Nursery Plants Research Unit } National Arboretum } } Bldg. 010A R.238 } 10300 Baltimore Avenue } Beltsville, MD USA20705 } } Phone: (301) 504-6097 } Fax : (301) 504-5096 } Email: brannign-at-asrr.arsusda. gov } } } }
Im looking for a functioning TN5500 with a Dec PC preferable an 1123 or better without the Detector for a project I'm working on. Has any of you recently upgraded thier probes and is looking to sell or donate it. I will gladly pay all associated expenses. Please contact me directly at the following: Regards,
Joe Barney Micro-Analytical Service Center Inc. Phone: 717-299-0599 Fax: 717-299-2022 E-mail: Jbarjr-at-aol.com
One thing which I've found to be helpful for getting rid of non-specific background labeling is 2-3 washes with a high-salt buffer immediately following the primary Ab. I've done quite a bit of work in the past with biotinylated primaries followed by streptavidin-gold, and have seen cases of high background on occasion; it somehow seems to be primary Ab-dependent...very bizarre.
Anyway, I'm wondering if something similar might be happening in your system. If you want to give it a try, I would suggest the following:
1. Make a buffer (TRIS HCl should be fine) containing 5 normal saline. Regular normal saline is 0.9% by weight, or about 150 mM, so add enough NaCl to make it 4.5% by weight (about 750 mM). I know this sounds like a whopping dose of salt, and I can't give a complete rationale for why it works, but it *does* decrease the background!
2. Do 2-3 washes w/ the high salt following the primary, then a couple of washes w/ the normal TRIS buffer to get the ionic concentration back where it should be for the gold conjugate.
3. One other thing: Do all of your diluents contain normal goat serum (even the diluent for the gold)? If not, you might want to try this as well.
Peggy Brannigan writes: } } (I'm labeling virus-infected plant leaf tissue with monoclonal antibodies } We block before primary antibody } incubation with a TRIS-HCl buffer containing bovine serum albumin and goat } normal serum.
Peggy, we had similar problems using some home grown antisera. Our solution was to block in 2%BSA, 5% Normal X serum and 1% Skim milk powder in TBS for 1 hr. Didnt completely solve the cell wall binding but was a *huge* improvment, didnt affect the specific binding at all.
Worth a try at least --
Daryl Webb (dwebb-at-waite.adelaide.edu.au) Dept. of Plant Science, Waite Institute University of Adelaide, Glen Osmond S.A. 5064 Australia. Voice:61_8 8303 7426 Fax:61_8 8303 7102
Many thanks to everybody who replied to my query about flatbed scanners. You are all a font of all kinds of information! To summarize:
1) (Obviously) get the highest res scanner with the most pixel depth you can afford - it's hard to overdo.
2) Printers that are 300-600 dpi with dither and print grey levels at 100-200 dpi; for continuous tome they need 3x3, 4x4, or 9x9 points (depending on the type of printer) for each pixel. So the higher the resolution of the image scanned in the better the print (also obvious).
3) Opinions vary - some say that 24 bit color depth gives 256 grey levels which is "more than enough", while others say that 30 or 36 bit depth gives more contrast enhancement, even when your software only handles 24 bit color. The best bet is to try out 24 and 30 bit scanners and see which does the job for you.
4) The big problem lies in enlarging images. A 4x5 SEM negative scanned in at 300 dpi can really only become a 4x5 print. A TEM negative scanned in at 600 dpi can be enlarged {3x, whereas a TEM negative can be enlarged 10x photographically. This brings up the discussion at MSA '96 - learn to take your micrographs at the magnification at which you will want to use and reproduce them rather than enlarging and therefore gaining only "empty magnification".
5) Remember to check for the true resolution, not the interpolated res. of a scanner.
6) Be careful of getting ghosts from glossy Polaroids and Newton rings from TEM negatives on some scanners.
7) None of the flatbed scanners seem good enough for 35mm negatives - buy a film scanner instead.
8) Remember that higher res images mean larger files. WIll you be able to store them? You will soon be needing ZIPs, JAZs, and CD-Rs. (Our SEM students are alloted 10 MB storage space on their department accounts. The full-size, full-res, colorized Mexican Ant on my web site is over 8 MB by itself!)
9) See "Tips & Tricks" at http://www.biotech.ufl.edu/~emcl/ for a previous discussion on this topic.
As you can see from my "cheap frame grabber" and "cheap scanner" posts, we are trying to operate within an embarrassingly small budget this year. ALthough I am still fairly happy to prepare everything photographically for myself, when I talk about image presentation and look at the bright shining faces of my current SEM students, I know that I am not going to be teaching them darkroom techniques. They start with Polaroid 55 pos/neg film. One needs to make slides right away; another wants to know how to make camera ready illustrations for a journal by next week. Most of these students ultimately have access to some decent hardware and software, and I need to point them in the right direction. I need about $10K to bring my lab up to speed. Too bad I can't invite you all to a huli-huli chicken sale!
What brought this up? Microtek 300 dpi scanners are down to $195, which I could buy out of my own pocket. It was tempting.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
What we need now is some advice for coating ceramic samples When to use Carbon, Gold, thickness etc.
Is there any FAQ about this available?
Any help highy appreciated.
______________________________________________________________ Andreas Loewe Tel: +49-228-734-180 University of Bonn Fax: +49-228-734-205 Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
your problem might be quite fundamental. It's several years since I've been seriously involved with immunogold labelling but I recall that there is a basic problem with monoclonal antibodies.
If the antigenic site, which the monoclonal antibody is specific to, is particularly sensitive to fixation then it will not label, hence the lack of specific labelling. You mentioned that a polyclonal works and this could simply be because it can 'find' a less sensitive part of the antigen. Have you had a chance to try cryo or less fixation?
I apologise if my comments are out of date or irrelevant.
Malcolm Haswell University of Sunderland UK ----------
Hello immunogold experts!
Hope I'm not duplicating an earlier topic but couldn't find anything in postings kept over the last few months so, here goes...
What factor (s) in an immungold labeling protocol has (have) the greatest impact on background, e.g.. antibody dilution, buffer, etc.? I'm looking at the immunogold protocol itself so I can use valuable tissue already embedded.
(I'm labeling virus-infected plant leaf tissue with monoclonal antibodies to viral proteins - samples are embedded in LR White and fixed in 3% paraformaldehyde/0.2% glutaraldehyde. We block before primary antibody incubation with a TRIS-HCl buffer containing bovine serum albumin and goat normal serum. Our secondary antibody is a commercially prepared goat anti-mouse gold conjugate. Usually we have excessive background on chloroplasts, mitochondria and cell walls; if we get rid of background by increasing dilutions of antibodies, we lose specific reactions as well).
One interesting point....we don't have this problem with polyclonal antibodies, only monoclonals.
Thanks for any tips -- this is really frustrating!
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
For x-ray analysis work use carbon (evaporate). It will (typically) attenuate less than Au and will not generate interfering x-ray lines. Because of the low Z coating and usual low z of ceramics, imaging of carbon coated ceramics should be done with as low a beam voltages as practical.
For the best imaging, use Au (sputter). It has better stopping power (= better resolution) and will provide a higher SE yield than carbon. Lower beam voltages (ie: 5-10 kv) still helpful.
Thickness: Always use the thinest coating which will prevent charging. This can sometimes be a problem on low density, friable ceramic fractures or fiberous material as a continuously conductive films can be difficult to achieve. One way to mimimize this is to paint as much of the specimen as possible (those areas not viewed) with carbon paint. Wicking can be a problem - viscosity and volitility of the paint will make quite a difference.
Alternatives (which I do not have available with my system)....
Very low kV imaging....in the area of approx. 500 to 1500 beam volts, some materials will be at an equilibrium state where energy in = energy out and thus dont't charge.
E-SEM - High chamber pressure discharges specimen and coating is not
needed. It has been my observation that this will work well for some
applications, but is surely no "cure-all". High mag, high resolution, low kV exams usually suffer.
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Dear folks,
we recently got a new SEM.
What we need now is some advice for coating ceramic samples When to use Carbon, Gold, thickness etc.
Is there any FAQ about this available?
Any help highy appreciated.
______________________________________________________________ Andreas Loewe Tel: +49-228-734-180 University of Bonn Fax: +49-228-734-205 Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
What we need now is some advice for coating ceramic samples When to use Carbon, Gold, thickness etc.
Andreas, it will depend ,to some extent, on what you are after. If you intend to do EDS for example, then you might want to deposit C since Au could interfere with your results. Also keep in mind that Au will deposit as relatively large islands and this will limit your resolution. You will be better off using Au-Pd for example as opposed to just Au. How thick a film you need will also depend , among other things, on the voltage that you are using in the SEM and on how flat your sample is (i.e. are you looking at a fracture surface or a polished surface ?). In most cases, at 10 or 20KeV , 10-30 nm of Au-Pd should be enough provided the sample is grounded properly (i.e. use a conductive paint, such as gold or carbon, around your sample). If you operate at lower voltages , a thinner coating might be O.K. You might want to read some standard texts on SEM and experiment a bit with your samples.
Jordi Marti ______________________________________________________________ Andreas Loewe Tel: +49-228-734-180 University of Bonn Fax: +49-228-734-205 Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear folks, } } we recently got a new SEM. } } What we need now is some advice for coating ceramic samples } When to use Carbon, Gold, thickness etc. } } Is there any FAQ about this available? } } Any help highy appreciated. } } ______________________________________________________________ } Andreas Loewe Tel: +49-228-734-180 } University of Bonn Fax: +49-228-734-205 } Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de } Inorganic Material Research } Roemerstr. 164 } 53117 Bonn } Germany http://www.elmi.uni-bonn.de/ } ______________________________________________________________ } } Andreas, First make sure that the sample is well grounded. Then it will be trial and error as to how thick to coat the sample. Gold usually gives the best results.--Good Luck
} Gregory Rudomen Greg-at-UMIC.SUNYSB.EDU University Microscopy Imaging Center S.U.N.Y. Stony Brook
The undesired signals are present only when you include your monclonal antibody, if I understand correctly. I don't think that it is just the fact that you have a monoclonal, after all, antiserum is more or less merely a collection of monoclonals.
If this undesired signal is caused by an unspecific reaction at the primary antibody level, as often found e.g. with IgM type antibodies (the bigger the stickier), you would expect to be able to get rid of it in the firat place by improving the block step. This takes care of background caused by hydrophobic interactions. Secondly, the incubation buffer should have additives (like BSA, cold water fish gelatin and there is a lot more!) which compete with primary antibodies for unspecific binding. In that case the protein additive should prevent or at least reduce background. Sometimes Tween does help, but since it is a detergent I would only use it on embedded material, not on cryosections. If you have to use detergent the best results are obtained at concentrations slightly higher than the critical micelle concentration.
On the other hand, from your description I get the impression that with increasing the antibody dilution both your specific and undesired signal decrease, which may indicate that "both types" of reaction have similar affinities. The question now is: is this undesired signal background or a more or less specific signal? Your specific reaction should at least have a lower Kd than the background reactions. If that is not the case, only a different antibody with higher affinity will help.
BTW: The topic of specificity and background will be extensively dealt with at the immunocytochemistry workshop in Iowa City, announced earlier on this Listserver.
Good luck!
============================= Jan Leunissen, Ph.D. AURION ImmunoGold Reagents & Accessories Costerweg 5, 6702 AA Wageningen The Netherlands
In 1992 Lindley (Microsc. Res. & Techn. 21:355) published a technique for using a primer (Z-6040, a silane) to allow for good adherence of biological materials with outer surfaces that do not normally stick well to embedding plastics. The plastic used was LRWhite. I am interested in using this primer with Epon 812 substitutes. Has anyone used this primer with the epoxies and is there a protocol.
In a similar vein, does anyone have a good protocol for embedding cell cultures on transwell membranes with small pores, such that when sectioning the cells + membrane the membrane does not rip away from the cells tearing off the basal layer of the bottom cells. Different membrane polymer types differ have different effects, as well as different pore sizes. If a very thick collagen layer is laid on top of the membrane this helps, but for experimental work this is often undesirable.
I just had to do a job for an investigator that wanted their prints made into slides. I have usually let them them take care of that and found out that our Biomed's graphic people use color slide film only which heard gives a blue cast to the EM prints. I decided to try Kodak's RPC # 175-3151 film on our copy stand (the people I got it from use it for taking slides of X-rays). It turned out Great! The draw back.... 10 and 15 second exposure brackets, at f4 f-stop, with four 500 watt photofloods. I developed it in our Xomat x-ray processor but was told you can use D-19 also. I'm going to reshoot the prints with Fugichrome 100 and that Direct MP 5360 someone else mentioned to compare. Has anyone else done this comparison? My next trick is to find out how to get the Poloroid Digital Pallete we have to use these long exposure films, if possible.
Thanks to all who responded to my cry for help recently re: what I mistakenly was calling moire patterns on images scanned using my Agfa Arcus II scanners.
Turns out, as was recently mentioned in a posting by Tina Calvaho (sorry if I got that wrong Tina), that the the effect is attributed to "Newton Rings" by those in the biz. The problem shows up when scanning negs which don't have very much detail.
The primary culprit in my case was a misalignment of the transparency module (ie the lid was on crooked). The effect was most prominent on the scanner with the greatest misalignment. This is not something which you can adjust unfortunately (holes in the chasis which accept the mounting brackets were slightly off), but Agfa was quick to send me another unit with it's head on straight.
None of my units are "perfect", but I'm told that something called "Newton Ring Spray" or matte spray solves any remaining problem. I'm hesitant to spray something on the glass, but if it's removable, I'll likely give it a try.
Cheers,
**************************************** Don Steele Steele-at-KRDC.INT.Alcan.Ca Alcan International Kingston Research and Development Centre (613) 541 - 2145 ****************************************
} comparison? My next trick is to find out how to get the Poloroid Digital } Pallete we have to use these long exposure films, if possible.
Rick brings up a point that should be considered when evaluating a scanner. Not only should you consider the pixel depth (8, 10 or 12 bits), but you should also consider the optical density range of the scanner. I don't think the recent thread on scanners touched on this.
The digitization depth basically indicates the *output* range of the scanner. A 10 bit scanner will output ~1000 gray levels. The optical density range, though, indicates the *input* range of your scanner. Optical density (OD) is the log(10) of the fraction of transmitted light. Since most scanners start at roughly an OD of 0, a scanner with an OD range of 2 will be able to "see" down to about 1% light transmission. An OD range of 3 provides you with a range of 1000 in the light transmission. With the same number of bits, each step will be 10 times coarser. You will, however, be able to pull information from those dark regions of the negative. Consider what kind of negatives you will be scanning. If they are relatively flat, an OD range of 2 may be sufficient. If you have high contrast negatives, check whether you need the extra input range.
The Umax scanner I've used has an optical density range of ~2.0. Agfa indicates that their Arcus II (US$1500-2000) has a density range of 3.0 (0.2 - 3.2). Their DuoScan ($4000-4500) has an OD range of 3.3 (0.2 - 3.5?). Polaroid's Sprintscan 45 apparently has an OD range of 3.4 with a $10k list price (ouch!).
So... think about your OD needs as well as pixel density, digitization bits, and price!
Cheers, Henk
Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility ------------------------------------------------------------------- Prosperity is the blessing of the Old Testament; Adversity is the blessing of the New. Francis Bacon, "Of Adversity."
Other than toluidine blue, as you mentioned, we also use a variation on the hematoxylin theme for our epon sections. We use Heidenhain hematoxylin-iron which comes in two solutions (I have the recipes if you want to make your own) - an iron alum solution and a hematoxylin solution. Slides are preheated and stained on a hot plate that is maintained between 80 and 90 degrees C. Coat the sections with the iron alum solution for 2-10 min depending on the tissue type, thickness, etc. Rinse VERY WELL with distilled water. Repeat the process on the hot plate with the hematoxylin staining solution using the same time interval as the iron alum. Because the "staining" only appears at this step, you may need to test a few slides to determine ideal timing. After rinsing well again with distilled water, flood the slides on the hot plate with TAP water to differentiate and let sit for 3 min. Rinse briefly in distilled water and dry. Prestaining like this allows the slides to then go through autoradiography without interfering with the emulsion and has worked for us irregardless of the fixative used.
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
I am looking for some one who as any information on something called antistatic "Denkil SEM" (Hodogaya Chemical Co. Ltd.). It was cited in a short article in J. Electron Microsc, Vol 28, No. 4 P 312-313.
Hoping to spark some ideas I have included the article, sic. (I apologize for any inconvience for members who are charged by file size basis, but its quite short:). Please, excuse any typos as I am not a professional typist, but I haven't altered any wording. Sorry I haven'y included the figures (but they do look impressive enough to bother trying to follow up)
----+----+----+----+----+----+----+----
J. Electron Microscopy, 1979, Vol. 28, No.4, 312-313.
Scanning Electron Microscopy of Lily Pollen Grains treated with Antistatic Solution
Masaru Katoh
(Nissei Sangyo Co. Ltd.)
Pollen grains are generally observed by SEM after drying in air and coating with metal. in some special pollens or in studies with higher resolution and magnification, however, routine procedures for preparation of general biological materials may be necessary.
In this letter, effects of acohol-soluble antistatic agent (1) upon electroconductivity of the specimen of lily pollen grains are introduced. in this method, specimens do not need troublesome metal coating and preserve much more natural shapes.
Figure 1a shows pollen grains of lily simply dispersed on a small piece of adhesive tape without following procedure of metal coating. in this micrograph, a trouble of charging up of the specimen is recognized. figure 1b is a micrograph from the same area as shown in Fig. 1a, but was taken after the treatment that the specimen drawn out from the microscope was immersed in several drops of 3% alcohol solution of the antistatic "Denkil-SEM" (Hodogaya Chemical Co. Ltd., Japan), and throughly dried in air. by this treatment, charging up of the specimen was completely eliminated. Another important effect of this method is that the pollen grains are expanded and the net pattern of the surface structure is restored. This may be a big advantage of this method keeping the pollen grains closer to the natural shape. This effect is probably due to a strong affinity of the antistatic agent with water restoring the orginal shapes of the pollen grains. Effective water may be derived from the dissolving component in alcohol and humid atmosphere. the absorbed water is probably retained firmly in the pollen grains even after exposuring to electron beam in a high vacuum.
Such an electroconductive treatment using water-retainable antistatic agent is considered very useful not only for prevention of charging up of specimens, but also, to some extent, for keeping some specimens moist even in a high vacuum.
Further examples of application of this antistatic agent will be reported in the near future together with many other interesting results.
Reference:
(1) Katoh, M. : J. Electron Microsc., 28, 51 (1979)
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That's it thats the entire paper. Does anyone out there have any ideas to point us in the right direction?
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
I've noticed a great deal of discussion regarding films for slides, and given how important this topic is to microscopists, I thought I'd add my experience to the list. My original film was also the MP 5360. The film works well, is of high resolution, and exposure times of one second or less using a MP-4 copy stand are common. You can process it in your own lab. The drawback, in my opinion, is a yellowish-brown hugh in the background. I am told this can be cured with fresh bulbs in the MP4 copy stand, and that they should be replaced "routinely", but I really do not want to be bothered with that responsibility and expense. I have also used the polaroid instant slide film, but I find the resolution unacceptable, and the slides I have made and archived do not seem to hold up to the test of time. Our current solution? We use Kodak Ektachrome 64T ("T" for tungsten lighting), EPY 135-36 film. The resolution is top shelf, and exposure times are on the order of 1 second or less. Standard E-6 processing at the local film processing stand does a nice job, so you don't even have to get your hands wet. On a 35mm camera set up on the MP4 copy stand, we use a film speed of "50", and f-stop of 2.8, and automatic exposure. I do not know the specifics about the lens which we use, but the distance to the camera is from about 8 inches to 2.5 feet, depending on the enlargement desired. With this film you will also experience a brownish background. Using a SLIGHTLY blue filter over the camera lens (available at the camera shop), a pleasing grey background can be gained. As a darker blue lens is used, the slides will become increasingly blue, but I for one prefer blue over brown.
Hope this helps,
Doug Keene Shriners Hospital for Children Portland, Oregon ---------------------- Doug Keene DRK-at-shcc.org
I recently purchased the Tektronics Phaser 550 colour laserwriter with extended features (1200 dpi) at great cost - ~$12K (OZ$). It arrived last week to great joy but test prints showed a regular fine horizontal line repeated at a spacing of 26.1mm when printing at premium grade. The spacing is 35mm at all other grades. The line is ~0.1-2 mm in width. It occurs in all images, ie colour, B/W and on test colour stripe images, ie it is independant of image type or source.
Complaint led to a rapid visit by the service engineer who confirmed it as a problem and that a second complaint had been lodged from a similar new installation several days earlier.
The Company response is now - that's laserprinters! IE the Tektronics Phaser 550 will always produce this regular and quality destroying line. I agree it is not strong but once noticed it catches the eye.
My query is:
has anyone else observed this regular lining?
Has anyone got a Phaser 550 which does not produce the lines?
I feel that this is totally unacceptable, needless to say these lines were not evident in sample prints, but the overall quality is good and the media print pricing is also good.
Note I am aware of dye-sub etc, I was after a unit with a lower media unit cost.
Thank you
Brendon J. Griffin Centre for Microscopy and Microanalysis The University of Western Australia Nedlands, WA, AUSTRALIA 6907 ph 61-9-380-2739 fax 61-9-380-1087
*Please note change to email address: bjg-at-cyllene.uwa.edu.au
Dear Andreas, The simple answer is: 1.) Use carbon evapoative coating when you want to do an EDX analysis of the ceramic. The thickness of carbon when a polished brass disk beside your sample just turns blue is 25 nm and is adequate for most samples. 2.) Use sputtered gold or sputtered 60%gold - 40%palladium alloy when imaging is your goal. Images are better when the sample is gold sputtered. A 10 nm coat is good. Your sputter coater should have a time vs. coating thickness chart. You wrote: } we recently got a new SEM. } } What we need now is some advice for coating ceramic samples } When to use Carbon, Gold, thickness etc. } } Is there any FAQ about this available? } } Any help highy appreciated. Good luck with your new SEM. Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Job vcancy for PhD student / Research Cooperation within area of biomimetic chemistry
We are looking for a PhD student to work on a project focusing on " Chemical reactions at microstructured surfaces ". The project is in cooperation with an industrial partner and the Central German Research Agency ( Leitz Electron Optics , LEO ) and focuses on the - chemical surface modification by microprinting techniques as published by Whitesites et. al. - study of surface reactions (for example formation of inorganic phases at patterned surfaces) within patterned surfaces in order to create spatially defined product areas. The work follows the ideas of biomimetic reactions at organic (polymeric) surface layers.
The structural part of the work focuses on the structural characterisation of surfaces and the reaction products formed at the surface. This should be done by microscopic methods inlcuding high resolution field-emisson scanning electron microscopy as well as Energy spectroscopic imaging and EELS analysis.
Experience in the surface modification by micropatterning is already available.
The project lasts till 31.12.1999
A cooperation with groups engaged in surface reactions, biomimetical reactions could also be possible. (Exchange of PhD students within the frame of this project).
For further information contact :
Dr. H.-G. Braun Institute of Polymer Research Dresden Microstructure Groups D-01069 Dresden Hohe Str. 6 Germany
We have experienced the same with a Lexmark Optra R at the higher dpi settings. Company says that it will always be a problem because the manufacturing process allows a rather high tolerance in the production of the gearing leading to chatter. They have given us two other machines which show the same problems. Needless to say we are quite dissatisfied with the product.
At 11:37 AM 3/20/97 +0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Hello Everyone, There has been a tremendous response to my plea for information about the DPM 2468. At the request of a couple of you, I am posting my findings to the list. Here is basically what I have learned from you folks, Kodak, and a couple of dealers with whom I have spoken.
1. Kodak is still making the DPM 2468. They won't produce the film, however, until they have an order for it.
2. Kodak will not sell this film in quantities less than 50 100ft. rolls. This makes dealers reluctant to stock it.
3. Neelima Shah informed me that Mid City Camera in Philadelphia carried this film and I was able to order 2 rolls from them. However, they are not sure that they will continue to stock it and are going to call me later this week to let me know what they decide. The 2 rolls they sold to me were the last they had in stock.
Now for the good news: Several of you have informed me of another film which seems to be very similar to this one in ease of use, exposure times and development. It is called Direct MP Film 5360 and is available from Ted Pella. You can find it on page 186 of their catalog. Those of you who have used this film seem to be pleased with the product. I would like to thank all of you who responded to my post on this listserver and for the information you shared with me. I attempted to answer each of you individually; but because the response became so great, I will have to thank you as a group. Once again you were there and willing to help. I am not affiliated with any of the organizations who make or sell any of the products discussed here. Again, thanks to all who shared. Sandra
Sandra F. Zane, EM Tech. sfzane-at-email.uncc.edu Dept. of Biology, UNCC Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
I am currently trying to switch from Epon embedding media to one which polymerizes by UV at lower temperatures. In order to orient my tissue properly I would like to continue using flat embedding molds. My problem is in making these molds "airtight" in order to prevent shrinkage/evaporation while it polymerizes. Has anyone had this problem or found a solution to it?
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
I have a Voyager EDX system. I want to perform a standardless semi-quant analysis on a solid solution. The solid solution is a combination of Barium, Strontium and Calcium carbonates, (BaCO3, SrCO3 & CaCO3). Can I have the Voyager look at the Ba, Sr & Ca and tell it that these exist as carbonates and have it calculate the weight percentage of each from me entering in the stoichiometry of the solution? Are there other suggestions for me to go about measuring the weight percentages of Barium carbonate, strontium carbonate and calcium carbonate in a solid solution?
} Date: Thu, 20 Mar 1997 16:32:07 } To: pat hales {hales-at-medcor.mcgill.ca} } From: Scott Whittaker {sdw-at-biotech.ufl.edu} } Subject: Re: UV Embedding in Flat Molds } } } } A procedure we use around here successfully is a piece of cleared film ( or aclar) punched with a hole puncher and placed on a pre-polymerized resin surface in a centrifuge tube. Just drop in the film, add tissue, add the resin, and polymerize as you would normally. } After curing, remove the tube and break the sample at the disk level. The resin doesn't stick to the film (or aclar) and you are left with the tissue at the surface of the block and ready to section in any orientation. Good luck } } } } } } } At 02:50 PM 3/20/97 -0800, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
The Pacific Northwest Electron Microscopy Society wishes to extend a warm welcome to all who wish to attend the Spring '97 Meeting at the V.A.Medical Center in Portland, Oregon Friday May 2nd, 1997. There are still a few spots for the up and coming young investigators/ graduate students to present papers on their current work to an enthusiastic Northwest audience of colleagues. Please send an e-mail to Charlie Meshul, Program chairman at meshulc-at-ohsu.edu with your topic if you wish to added on. Alternatively send e-mail to Bob Kayton kayton-at-ohsu.edu or Bob Mixon mixonr-at-osu.edu
Has anyone hired a commercial enterprise to conjugate their primary antibody to a gold particulate? We have a couple of mouse monoclonals that we would like to simultaneously localize in tissue, but do not have the time or experience to do the conjugations ourselves.
Many thanks,
Doug Keene Associate Investigator Shriners Hospital for Children
Greetings, For flat embedding under uv light without air, we use flat bottom capsules. These are made by TAAB in UK and distributed by several USA em suppliers. If you have really big samples, I beieve the little plastic widgets that em grids come in will work and these too are available from suppliers (EMS at least, and no doubt others too). Hope this helps,
I have hundreds of 35mm color negs which I wish to make proof sheets of. I do not want to do anything with those images other than print proof sheets. Can anyone suggest a good scanner for this job.
I have a PhD student from Brazil who is studying Catolaccus grandis. He places a female on top of a beeswax cell which contains boll weevil larva. The female deposits an egg on the larva and 2 weeks later, it matures and burrows its way out of the beeswax creating a hole. He would like to study and measure these holes.
However, from past experience I know that beeswax melts when placed in my Au/Pd sputter coater. Is there any way to coat the beeswax and examine them with an SEM? Or should he try another route?
Thank you in advance,
Ginger Baker EM Lab Manager Dept. Anatomy, Pathology, and Pharmacology 250 Veterinary Medicine Oklahoma State University Stillwater, OK 74078 (405) 744-6765 FAX: (405) 744-5275 Email: lizard-at-okway.okstate.edu
Regarding your question about flat embedding samples in media which polymerize under U.V.. We have used both LR White and Lowicryl resins successfully for flat embedding. We use Beem flat embedding molds, which are quite stiff and seem to contain less free oxygen than standard molds. Our trick to exclude atmospheric oxygen is to use the plastic pipette tip box that "Multi-Flex" tips are packaged in. It is a three part box which consists of a UV light penetrable top, which fits over a multi-holed mid section (normally supporting the tips) which fits over a lower compartment. I collected a bunch of these from another lab, but I do not know where to get them. We fill the lower compartment with dry ice (solid CO2), and drill a couple of holes in the top (letting CO2 gas escape as the solid sublimes). The samples in the beem flat embedding molds are placed on the perforated "shelf" above the CO2 chamber and exposed to a constantly replenished environment of gaseous CO2, excluding incoming oxygen. We put the whole thing, including a UV light, in the -20 portion of our freezer. The polymerization proceeds fully enough by the time the dry ice is gone to prevent oxygen inhibition. You might also try using a "heat sink" type of mold. Ted Pella sells one using a teflon mold which slides over aluminum ( I have no commercial interest in Pella).
You didn't mention what kind of UV-polymerizing resin you'll be using, but if it is Lowicryl or a similar methacrylate, here's how to do it:
1. Use polyethylene flat embedding molds. These molds are made from the same type of material as BEEM (R) capsules. The polyethylene molds are transparent to the UV light, making the polymerization much more even. Get back to me if you can't find a vendor for the polyethylene molds, I can't remember at this moment who sells them -- Ted Pella, I think...but I can look it up for you.
2. Place the mold on a piece of cardboard that is wrapped in aluminum foil. The cardboard should be larger than the mold. This makes moving and transferring the mold much easier and less messy (see below).
3. Overfill all of the cavities in the mold to give a "positive meniscus" at each position. If some of the cavities don't have specimens in them, go ahead and fill the cavities anyway.
4. Cut a piece of Parafilm (R) that is slightly larger than the mold. Then, beginning at one end of the mold, gently lay the Parafilm down on top of the mold. As you do this, the excess resin will run out of the mold cavities and get trapped under the Parafilm. Gradually lower the Parafilm down toward the other end of the mold, allowing the resin to fill the space between the mold and the Parafilm. This seals the complete top of the mold. The resin won't dissolve the Parafilm, but it will make it soft.
5. You can use tissues to absorb the excess resin as it runs down the outside of the mold. (**CAUTION:** Be sure to wear gloves at this step, and at all steps when working with methacrylate resins -- or any resin, for that matter!).
6. Transfer the filled molds to your polymerization apparatus. I don't know what you will be using, but I make polymerization chambers out of cardboard boxes. Just make two cutouts: one on the top for the UV lamp, and one on the side for a door. **To make the polymerization even, cover all of the inner surfaces of the box with aluminum foil.**
7. **NOTE**: You can also use Thermanox (plastic) cover slips instead of Parafilm in Step #4. Just lay the coverslips on top of the cavities, letting the excess resin run out along the edges.
8. Let everything equilibrate at low temperature for 15-20 minutes, turn on the UV lamp, and you're on your way to polymerized resin!
9. **NOTE**: If you see "bubbles" or vortex-like "swirls" in the polymerized blocks, this is usually a sign that the polymerization was too rapid. Increase the distance between the lamp and the mold to cut down on the UV intensity. If this isn't possible, place a piece of frosted glass between the lamp and the molds.
I've never asked anyone to conjugate a monoclonal Ab directly to colloidal gold, mainly because of the added cost (a similar concern of yours, I take it!).
Are you up for a slightly different approach, which you can do with commercially available reagents? If so, you could do the following:
1. On one side of the grid (maybe a fairly high hexagonal mesh, uncoated nickel grid?) incubate with one of your mouse monoclonals.
2. Follow with biotinylated goat anti-mouse Ig-whatever (is it an IgG?).
3. Follow with streptavidin-gold (available in many different sizes).
4. Flip the grid over and repeat the labeling with the second monoclonal. Only this time, use a different size of streptavidin-gold.
5. Double sided labeling works quite nicely for co-localizations, and it eliminates a lot of cross-reactivity problems you'd have to fight if you worked on only one surface of the section.
6. One thing to watch out for: If you add normal goat serum and maybe a little TWEEN or other additives to the reagents, the grids will tend to sink if they're incubated on drops of the reagents. Then you're labeling *both* sides with the *same* reagents. This, of course, is to be avoided! You have to keep the sides separated.
The best way to avoid this problem is to anchor the grid by its edge on a piece of double-stick tape on a microscope slide. You can then use a micropipette to deliver *very small* volumes (maybe 2-3 microliters) of each reagent. To change reagents or to wash, simply add the reagent to one edge of the grid and wick it off with filter paper from the opposite edge.
Following the first gold incubation you can remove the grid and wash well in a gentle stream or w/ repeated drops of buffer and finally water. Allow to dry, flip the grid over, stick it down to the tape and do the second run-through.
An added bonus of this procedure is a fairly good signal amplification. If you conjugated your primary directly to the gold, you'd only have a one-step reaction with minimal gold particles (probably 1) binding at each site. Would that be enough to detect above background?
Unless, of course, you *want* the most specific localization possible...In that case, forget everything I've just written!
I've never asked anyone to conjugate a monoclonal Ab directly to colloidal gold, mainly because of the added cost (a similar concern of yours, I take it!).
Are you up for a slightly different approach, which you can do with commercially available reagents? If so, you could do the following:
1. On one side of the grid (maybe a fairly high hexagonal mesh, uncoated nickel grid?) incubate with one of your mouse monoclonals.
2. Follow with biotinylated goat anti-mouse Ig-whatever (is it an IgG?).
3. Follow with streptavidin-gold (available in many different sizes).
4. Flip the grid over and repeat the labeling with the second monoclonal. Only this time, use a different size of streptavidin-gold.
5. Double sided labeling works quite nicely for co-localizations, and it eliminates a lot of cross-reactivity problems you'd have to fight if you worked on only one surface of the section.
6. One thing to watch out for: If you add normal goat serum and maybe a little TWEEN or other additives to the reagents, the grids will tend to sink if they're incubated on drops of the reagents. Then you're labeling *both* sides with the *same* reagents. This, of course, is to be avoided! You have to keep the sides separated.
The best way to avoid this problem is to anchor the grid by its edge on a piece of double-stick tape on a microscope slide. You can then use a micropipette to deliver *very small* volumes (maybe 2-3 microliters) of each reagent. To change reagents or to wash, simply add the reagent to one edge of the grid and wick it off with filter paper from the opposite edge.
Following the first gold incubation you can remove the grid and wash well in a gentle stream or w/ repeated drops of buffer and finally water. Allow to dry, flip the grid over, stick it down to the tape and do the second run-through.
An added bonus of this procedure is a fairly good signal amplification. If you conjugated your primary directly to the gold, you'd only have a one-step reaction with minimal gold particles (probably 1) binding at each site. Would that be enough to detect above background?
Unless, of course, you *want* the most specific localization possible...In that case, forget everything I've just written!
Pat Hale wrote: ================================================== I am currently trying to switch from Epon embedding media to one which polymerizes by UV at lower temperatures. In order to orient my tissue properly I would like to continue using flat embedding molds. My problem is in making these molds "airtight" in order to prevent shrinkage/evaporation while it polymerizes. Has anyone had this problem or found a solution to it? ================================================== There is an alternative to the rigid polyethylene based (e. g. BEEM manufactured) flat embedding molds, and that is the flexible and reusable UV transparent silicone flat embedding molds. These silicone molds work just fine and seem to have a bit higher transparency for the UV. One would follow the method described by Bob Chiovetti, that is, to overfill the cavities but instead of using Parafilm, just place the bottom of another one of the UV transparent silicone molds on top. Capillary action quickly seals out any entrapped air, creating the desired oxygen free environment.
UV transparent embedding molds come in varous sizes and shapes and are shown on the SPI website given below. Similar, but not idential molds are also offered by Ladd and possibly others.
Disclaimer: SPI Supplies manufactures UV transparent silicone embedding molds and has a vested interest in seeing more persons using them!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The Polaroid Sprint Scan 35LE will scan your negatives in at resolutions up to 1950 dpi -at- 10 bits per channel with an optical density of 3.0 in under 60 seconds. It will scan 35 mm negatives or slides(mounted or unmounted)as well as the SuperSize format. It has an option called the PathScan Enabler that allow you to scan a standard glass slide at the referenced resolution and color depth. It has a list price of $995US.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation
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I have hundreds of 35mm color negs which I wish to make proof sheets of. I do not want to do anything with those images other than print proof sheets. Can anyone suggest a good scanner for this job.
POSITION AVAILABLE!!!! At the Institute for Neurobiology, University of Amsterdam, The Netherlands a post-doc position is available, for 3 years. The project concerns plasticity in brain cells, in relation to epilepsy and hormonal influences. It will make use of (fast) calcium imaging techniques with confocal microscopy, in combination with patch clamp recording. We are looking for someone with experience in confocal microscopy, preferably in brain tissue. The position is available as of now. For more information: wadman-at-bio.uva.nl or joels-at-bio.uva.nl or call 31205257641 / 31205257626.
----------------- P.C.Diegenbach dept. Biology, University of Amsterdam Kruislaan 320 1098 SM Amsterdam, Netherlands phone (31) 20 5257631 fax (31) 20 5257709 The following email address is mangled to prevent automated unsolicited junk mail. Replace the '_AT_' with an '-at-': email paulcd_AT_bio.uva.nl -----------------
Pat: I use UV polymerization of LR white or Unicryl at 4C for immunolabelling studies. I use small aluminum weighing dish layered with a sheet of Aclar film at the bottom. The embedding and tissue are placed on top of this film. Cut a sheet of closely fit Aclar film to place over it. All the weighing dishes are placed in a stainless tray which is sealed with Saran Wrap. The aluminum will reflect the Uv and provide good even polymerization. The embedding medium between the sheet will polymerize while some excess media flow over and under the sheet will not. The Aclar sheets are easily peeled off from the polymerized material. Sheets of embedding material can be scanned under LM to chose for ideal orientationed sample. Hope this will help you.
Licy Yin Microscopy Center U.Mass , Amherst, MA 01003t MiclOn Thu, 20 Mar 1997, pat hales wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am currently trying to switch from Epon embedding media to one which } polymerizes by UV at lower temperatures. In order to orient my tissue } properly I would like to continue using flat embedding molds. My problem is } in making these molds "airtight" in order to prevent shrinkage/evaporation } while it polymerizes. Has anyone had this problem or found a solution to it? } } Pat Hales } McGill University } Dept. of Anatomy & Cell Biology } hales-at-hippo.medcor.mcgill.ca }
We have had superb results with Fuji's Velvia. It is a daylight film, so you will need blue filtration when used with tungsten halogen sources (82B)and it is slow (50ASA) but it produces exquisite color rendition and depth. It is also a standard E-6, for easy development. Good luck... and if you take an especially beautiful shot which you would like to share, let us know. We are always looking for interesting images for our short courses (with credit, of course).
Barbara Foster Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 (413)746-6931 fax: (413)746-9311 email:mme-at-map.com
Pat Hales asks about airtight seals for acrylic embedding medium.
Unicryl, an acrylic embedding medium, advertises that excluding oxygen during UV polym is not necessary. Unicryl is available from a number of vendors. (I have NO interest in the commercial uses of Unicryl). Using an acrylic which bypasses the entire problem of oxygen interference might be a good solution. Bye, Hildy
MIDWEST MICROSCOPY AND MICROANALYSIS SOCIETY affiliate of the Microscopy Society of America and the Microbeam Analysis Society
presents
WORKSHOP AND SYMPOSIUM ON IN SITU HYBRIDIZATION
The program has been arranged by and will be held at Madison Area Technical College (MATC), WI., Friday, April 11, 1996 from 8:00 - 5:00 p.m. This meeting will consist of two concurrently held workshops and an afternoon related symposium. The general emphasis will be in situ hybridization. The two morning workshops will be on detection of hybridization with radioactive and non radioactive probes.
The meeting has several purposes:
1. To draw the attention of the scientific community to emerging developments in the practical and basic research aspects of excitingnew fields.
2. To bring people together from diverse disciplines in order to discuss how innovative techniques will be relevant to the future direction of microscopy and microbeam analysis.
WORKSHOPS WILL BE HELD IN THE MORNING FROM 8:30 -12:30.
1. Radioactive in situ hybridization Patricia L. Allen/Dr. Lyn Thet Department of Pulmonary Medicine, School of Medicine UW-Madison
2. Nonradioactive in situ hybridization Boehringer Mannheim, Indianapolis, IN
You may choose to attend one workshop from above. Registration information is enclosed. Register early because we will have to limit the number of participants.
AFTERNOON SYMPOSIUM SCHEDULE 1:30 - 5:00 p.m.
Patricia Thomas, M.D. Department of Pathology, School of Medicine, University of Iowa, Iowa City, IA, "Diagnostic applications of fluorescence in situ hybridization"
Kathrine Staskus, Ph.D. Department of Microbiology, University of Minnesota, Minneapolis, MN, "In situ hybridization detection of Simian Immunodeficiency virus (SIV) proteins and nucleic acids"
Gary Lyons, Ph.D. Department of Anatomy and Cellular and Molecular Biology Program, School of Medicine, University of Wisconsin, Madison, "In situ hybridization studies of murine embryonic skeletal muscle development"
Alan Smith, PhD. Department of Horticulture, College of Agriculture, University of Minnesota, Minneapolis, MN, "Localization of floral specific gene products during pollen development"
Gwen V. Childs, Ph.D. Vice Chair, Department of Anatomy and Neuroscience, Program Director. Cell Biology Graduate Program, University of Texas Medical Branch, Galveston, TX, "Regulation of multipotential cells in the pituitary by neuroendrocrine peptide:an in situ hybridization and immunocytochemical study"
Kevin Roth, M.D., Ph.D Department of Pathology, Washington University, St. Louis, MO. "Application of tyromide-signal amplification to in situ and immunocytochemistry"
WORKSHOP AND SYMPOSIUM ON IN SITU HYBRIDIZATION
GENERAL INFORMATION SHEET
DATE: Friday, April 11, 1997
LOCATION: Madison Area Technical College (Truax Campus) Electron Microscopy Program (Check-in, Rm375A) 3550 Anderson St., Madison, WI. 53704
Workshops and Symposium locations will be provided at the Check-in room (Rm375A)
DIRECTIONS PARKING: A map is provided on request. Please park in the large lot on the side of the campus building.
MEALS: Refreshments will be provided in the morning and for afternoon break. Lunch is a sandwich buffet, and the cost is $4.00/person. Following the afternoon symposium there will be a wine, soft drink, and cheese reception provided at no cost to participants. An Italian dinner buffet is being organized for after the wine and cheese reception. The cost is $7.00/person.
REGISTRATION: If you plan to attend either one of the workshops or the symposium you must register by phone or EMAIL to one of the contacts listed below. Registration is free to all Society members, student, regular, and corporate. If you are not a member of the Society, registration cost will be the price of membership. (Regular = $10, Student = $5). When you register you must provide us with the information listed below. Name tags and Fees for registration and meals will be collected at the Check-in site.
DEADLINE DATE FOR REGISTRATION IS APRIL 8.
1. Name 2. Affiliation 3. Phone # 4. Choice of one Workshop 5. Are you attending the Afternoon Symposium? 6. Do you want to participate in the Lunch or Dinner?
REGISTRATION CONTACTS:
James P. DiOrio (847) 270-4676 Fax: (847) 270-4414 EMAIL: diorio-at-baxter.com
Joanne M. Crudele (847) 734-3712 Fax: (847) 734-3686 EMAIL: Joanne.Crudele-at-Unilever. Com
If you need information on hotels, contact Michael Kostrna (608)246-6762 or Kenneth Muse (608)243-4309.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello all, } } I have a PhD student from Brazil who is studying Catolaccus } grandis. He places a female on top of a beeswax cell which contains } boll weevil larva. The female deposits an egg on the larva and 2 } weeks later, it matures and burrows its way out of the beeswax } creating a hole. He would like to study and measure these holes. } } However, from past experience I know that beeswax melts when placed } in my Au/Pd sputter coater. Is there any way to coat the beeswax and } examine them with an SEM? Or should he try another route? } } Thank you in advance,
} Email: lizard-at-okway.okstate.edu } Do you have cooling system on the sputter coater ? If your answer is yes, let the beeswas sit on the cooling stage for 20 min then coat it for 5-10 sec and pause for about 20 sec. Repeat it for 3-4 cycles. My experience for fibric material the above tricks would work fine. Also you may lower your sample far from the gold target to avoid heat.
Hope this will help.
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
With a Special Lecture and Optional Demonstration Workshop on in situ Hybridization on Sunday, April 27 presented by Dr. L=E1szl=F3 K=F6m=FCves of UC San Francisco
Business meeting for CSU delegates, Friday, April 25, 1997 at 4:00 PM in Duncan Hall Rm. 249
Proceedings will be published in Microscopy Research and Technique
The Colloquium provides a forum for the exchange of research results and experiences in all fields of light and electron microscopy in the biological, geological, and materials sciences. Participants include students and scientists from academia and industry.
Both platform and poster presentations are invited. Student presentations are strongly encouraged.
Registration Fees $35 Regular, $10 Student, $50 Vendor by March 15; $25 Optional Workshop
Late Registration $45 Regular, $10 Student, $60 Vendor, after March 15; $25 Optional Workshop
Make checks payable to San Jose State University Foundation
Send to: Dr. David K. Bruck CSU Microscopy Colloquium Department of Biological Sciences San Jose State University San Jose, CA 95192-0100
=46or additional information, flyers, and abstract forms, contact David Bruck at phone: (408) 924-4837 or fax (408) 924-4840 bruck-at-biomail.sjsu.edu
Program
=46riday, April 25
4:00 PM Business meeting for CSU delegates in Duncan Hall Rm. 249, San Jose State University
Saturday, April 26
8:00 AM Registration, coffee, and breakfast snacks in lobby of Duncan Hall, San Jose State University 9:00 AM Opening remarks in Duncan Hall Rm. 135 9:15 AM Platform presentations 10:30 AM Coffee break 11:00 AM Platform presentations 12:00 PM Buffet lunch 1:30 PM Platform presentations 2:30 PM Coffee break 3:00 PM Platform presentations 4:00 PM Lecture on in situ hybridization by Dr. Laszlo Komuves 5:00 PM Closing Remarks and Poster session in Duncan Hall Rm. 250 6:30 PM Banquet
Sunday, April 27
9:00 AM Demonstration and workshop on in situ hybridization conducted by Dr. Laszlo Komuves in Duncan Hall Rm. 344
Housing
Best Western Inn 455 S. Second St. San Jose, CA 95113 phone (800) 528-1234 or (408) 298-3500 singles $52, doubles $62
Please make your own reservations for housing at one of the phone numbers above. Specify that you will be attending the Microscopy Colloquium to receive the conference discount. A map and other hotels in the vicinity will be included in the packet of information mailed to you following receipt of your registration information.
Transportation
=46ree parking on campus is available in the 7th St. Garage on the corner of 7th St. and San Salvador St. A permit will be mailed to you following receipt of your registration form.
Public transportation from the San Jose International Airport to downtown San Jose is limited to cabs or the Light Rail system. For the latter, a free shuttle bus (labeled Metro) runs every 10-15 minutes from the airport to the Metro Light Rail Station. For $1.10, the train runs downtown to the Paseo de San Antonio Station, about 3 blocks from the university and 2 blocks from the Best Western Inn.
Registration
Please complete the registration form below whether or not you will be presenting a paper or poster.
After March 15 $45 regular, $10 student, $60 vendor
_____________________________ Name
_____________________________ Address
_____________________________ City State Zip
_____________________________ Phone
_____________________________ e-mail
Workshop Registration (for an additional fee of $25) 0 Workshop
Note that the workshop's occurrence will depend on an adequate number of registrants.
Meal Preference 0 Chicken 0 Fish 0 Vegetarian
Presentation 0 Platform 0 Poster 0 None
Information and Mailing
Make registration checks payable to the San Jose State University =46oundation. Mail registration forms, abstract forms, and checks to
Dr. David K. Bruck CSU Microscopy Colloquium Department of Biological Sciences San Jose State University 1 Washington Square San Jose, CA 95192-0100
(408) 924-4837 bruck-at-biomail.sjsu.edu
Platform and Poster Presentations
Papers discussing any aspect of microscopy in cell, developmental, and structural biology or in the material or geological sciences are welcome. Microscopy techniques' papers are particularly encouraged.
Platform presentations (15 min.) will be held Saturday, April 26. Posters will be displayed all day Saturday, with an afternoon session designated for viewing with authors present.
An Abstract Guidelines form is included for those presenting papers or posters. The abstracts will be compiled into the "Proceedings of the Fifth Annual CSU Microscopy Colloquium" and will be published in Microscopy Research and Technique.
Dr. Steven Barlow EM Facility/Biology Department 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/EM_Facility
Carolina Workshop on: LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES June 1-6, 1997 University of North Carolina at Chapel Hill
Instructors: John J. Lemasters Edward D. Salmon Brian Herman ------------------------------------------------------------------- LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES is an introduction to applications of light microscopy. Students will have opportunities for extensive hands-on experience with state-of-the-art equipment for optical imaging, digital imaging processing, fluorescence microscopy and confocal microscopy guided by experienced academic and commercial staff. The course is divided into three major sections with lectures and laboratory exercises on: 1) geometric and wave optics of image formation, microscope alignment, phase contrast and reflection interference contrast microscopy; 2) video imaging, including contrast enhancement by analog and digital image processing, fluorescence microscopy, image detectors, fluorescent probes, ion imaging, and green fluorescent protein; and 3) laser scanning confocal microscopy emphasizing live cell imaging and 3-dimensional image reconstruction. Students are encouraged to bring their own specimens for analysis. A commercial staff representing leading microscopic manufacturers will make available for student use the latest and most advanced instrumentation for light microscopy, image detection and computerized image analysis.
Tuition is $950.
------------------------------------------------------------------- APPLICATION FORM Carolina Workshop on LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES
Position:
Address:
Telephone:
Fax:
Please return this form along with a brief letter describing your research interests and a curriculum vitae. Applicants should contact the program as soon as possible. Full consideration will be given to applications received by April 18, 1997.
Send application to: Dr. Wayne Litaker, Director of Workshops University of North Carolina at Chapel Hill Program in Molecular Biology & Biotechnology CB# 7100, 442 Taylor Hall Chapel Hill, North Carolina 27599-7100 Tel: (919) 966-1730 Fax: (919) 966-6821 e-mail: litaker-at-med.unc.edu ------------------------------------------------------------------- {End of Announcement}
I have one single question for the wealth of knowledge out there in the world of Microscopy....
On a Nikon Diaphot inverted with a universal Con.A Achr-Apl 1.4 condenser using a 10X objective with DIC do you have to oil the condenser????
Also how do you do the alignment properly, since noone in the lab I work knows... I think I have everything correct but when I twist polarizer by the objective lens I do not get the rainbow of colors or even the black background when the polarizers are ligned up....
Any Suggestions \\|// (o o) ~~~~~~~~~~~~oOOo~(_)~oOOo~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
{ { {This message is made of 100% recycled electrons} } }
Cheers ;o) :o) %o) Eric {Mesa Arizona} http://www.goodnet.com/~earosen (Note the tilde before earosen)
An alternative may be replication using Reprosil, a Polyvinyl Siloxane from Caulk. It makes a negative which can be transfered to a positive by using Spurr low viscosity embedding medium. Detail is there albeit a little softened.
I am a student now using Hitachi-8100 TEM to study submicrometer Cu-Zn alloy particals. The particals are mostly six-fold symmetry plates. When the beam focused on a partical at a high magnification, some dark stripes appear in the partical's bright-field image and change their shapes until a snow-flake-like pattern is formed. The pattern comprised of six center-crossed dark stripes. Select-area diffracton shows a single crystal diffracton pattern with six-fold symmetry. But dark-field image shows that one single diff spot is not coming from the whole partical but from ONE corresponding dark stripe.
My questions are: 1)Do anyone out there ever find similar phenomenon? 2)How can I know the temperature of the particals under the brightening of the electron beam? I think temperature is a reason for the formation of the snow-flake-like pattern.
I hope I have described my question clear enough for you to reply. Thanks!
Sun Haiping National Lab of Superhard Materials Jilin University Changchun 130023, P.R.China sunqa-at-mail.jlu.edu.cn
Aclar is a thin plastic sheet material suitable for cell culture and may be sectioned. A full description is in our catalogue on page L7. We distribute Aclar as does Ted Pella (Pelco) in the USA. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au
} Several of you have recommending using Aclar film in your UV polymerization } protocols. What is Aclar film, and where do you purchase it? } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX) } }
Hello, I am having no luck attempting to stain the glycocalyx? of a species of sulfate-reducing bacteria using ConA-conjugated gold (5nm). Using 4%PF+0.1%Glute in Cac buffer fixation followed by LRW embedding I have tried 1:10 to 1:100 in P04 buffer pH 7.2. I have also tried ConA-gold in Tris-saline with and without 1mM CaCl2 and 1mM MgCl2 at pH 6.9,7.2 and 8.0. diluted 1:10-1:100. Next I tried taking a fresh culture and washing several times in Tris-saline followed by 1:10 ConA-gold,then fixing and embedding. My incubations have been around 1 hr at room temp. Nary a gold particle do Isee under any of the conditions mentioned. Apparently, using fluorescently labelled ConA these little buggers lightup like the Hale-Bopp comet, so Iam at a lost. Can anybody out there give me some suggestions. I also have some material that I fixed in 4% PF alone, embedded in LRW that I have not done anything yet with. Thanks in advance
Hank Adams Electron Microscopy Laboratory New Mexico State University Las Cruces, NM 88003
I have an American Optical Micro-Star Illuminator model 1872. It has 10XB&L WF eyepieces and a third place for camera/eyepiece. The primary voltage is 115 and secondary 3 to 6 volts using GE lamp 1974 It has no objectives, but spaces for 4. It has a large mechanical base. It is illuminated thru the lenses.
I know nothing else about the scope except it was used in a lab at Fairchild semiconductor in South.
Iam looking for some good quality/inexpensive objectives for amatuer use and any manuals/info on using this scope.
If you know anything of this scope or the objectives that I need, please e-mail me
Thanks so much for your time
David Chase dchase-at-gwi.net
ps I had researched this all once before but a fatal HD crash destroyed all my data.
Hello, I am having no luck attempting to stain the glycocalyx? of a species of sulfate-reducing bacteria using ConA-conjugated gold (5nm). Using 4%PF+0.1%Glute in Cac buffer fixation followed by LRW embedding I have tried 1:10 to 1:100 in P04 buffer pH 7.2. I have also tried ConA-gold in Tris-saline with and without 1mM CaCl2 and 1mM MgCl2 at pH 6.9,7.2 and 8.0. diluted 1:10-1:100. Next I tried taking a fresh culture and washing several times in Tris-saline followed by 1:10 ConA-gold. My incubations have been around 1 hr at room temp. Nary a gold particle do I see under any of the conditions mentioned. Apparently, using fluorescently labelled ConA these little buggers light up like the Hale-Bopp comet, so I am at a lost. Can anybody out there give me some suggestions. I also have some material fixed in 4% PF alone, embedded in LRW that I have not done anything yet with. Thanks in advance,
Hank Adams Electron Microscopy Laboratory New Mexico State University Las Cruces, NM 88003
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Hank Adams writes: "Hello, I am having no luck attempting to stain the glycocalyx? of a species of sulfate-reducing bacteria using ConA-conjugated gold (5nm)." (trimmed).
Dear Hank, Your problem may lie in the ConA-conjugated gold. Often, these probes very seldom store well. If most of the protein has dissociated from the gold particles, you will have a large amount of very specific ConA binding but no gold to show you it is there. It is easy to test at the LM level by probing sections treated with ConA-god with anti-ConA and fluorescent antibody.
Once again, the best visualization method for lectins (re:ricin) may be to use antibodies instead of the directly conjugated gold probes. If you prefer to use ConA-gold then I recommend that you make your own. A slightly less satisfactory approach would be for you to centrifuge down your ConA-gold (be careful not to spin it so hard that is aggregates into the bottom of the tube) and resuspend it in fresh PBS. This will remove most of the free protein.
As an aside and in reference to a previous posting, streptavidin-gold also suffers from this storage problem. When using the probe at the LM level, with silver enhancement, or with multiple antibody layers the loss of labeling efficiency is not so noticeable. However, attempting to visualize biotinylated antibodies directly with streptavidin-gold will only work well for the first week after the probe has been made. For situations where there are large numbers of bound antibody, this effect may not be noticeable, but for situations where there are only low numbers of antigens, it is very obvious.
Making colloidal gold and coupling it to proteins or antibodies is easy and anyone with even limited experience can do it.
Check out my WWW "gold page" to see how easily it can be done. Be warned, if you want to do this you usually need large supplies of protein (a point anyone coupling antibodies should be aware of).
Paul Webster Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
Message-Id: {199703230345.UAA07299-at-mailque.goodnet.com} Comments: Authenticated sender is {earosen-at-pop.goodnet.com}
You may also want to treat the sections with a very low concentration of sodium periodate which will destroy the section if you are not careful.. This will expose some of the bnding sites fo the Con-A.. or on the other hand the Lctins are too old, and the gold has fallen off of the lectin which occured to me also when I was using gold labeled lectins that were stored in the fridge........ So I got very scattered labeling or no labelling at all but when used in teh LM with Cy3 they light up like a chrsitmas tree....
lAlso you may want to try a lower comcentration of fixatives in Formaldehyde and Glut.
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Hello, I am having no luck attempting to stain the glycocalyx? of a species of sulfate-reducing bacteria using ConA-conjugated gold (5nm). Using 4%PF+0.1%Glute in Cac buffer fixation followed by LRW embedding I have tried 1:10 to 1:100 in P04 buffer pH 7.2. I have also tried ConA-gold in Tris-saline with and without 1mM CaCl2 and 1mM MgCl2 at pH 6.9,7.2 and 8.0. diluted 1:10-1:100. Next I tried taking a fresh culture and washing several times in Tris-saline followed by 1:10 ConA-gold. My incubations have been around 1 hr at room temp. Nary a gold particle do I see under any of the conditions mentioned. Apparently, using fluorescently labelled ConA these little buggers light up like the Hale-Bopp comet, so I am at a lost. Can anybody out there give me some suggestions. I also have some material fixed in 4% PF alone, embedded in LRW that I have not done anything yet with. Thanks in advance,
Hank Adams Electron Microscopy Laboratory New Mexico State University Las Cruces, NM 88003
Can any one tell me of a Microscope news group taylored toward the newbie into this hobby? Also, are there any good magazines on the market specializing in MIcroscopes and the Microscope hobby?
} Date: Sat, 22 Mar 1997 15:20:12 -0500 } From: H. ADAMS {hadams-at-nmsu.edu} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Lectin staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, I am having no luck attempting to stain the glycocalyx? of a } species of sulfate-reducing bacteria using ConA-conjugated gold (5nm). } Using 4%PF+0.1%Glute in Cac buffer fixation followed by LRW embedding I } have tried 1:10 to 1:100 in P04 buffer pH 7.2. I have also tried ConA-gold } in Tris-saline with and without 1mM CaCl2 and 1mM MgCl2 at pH 6.9,7.2 } and 8.0. diluted 1:10-1:100. Next I tried taking a fresh culture and } washing several times in Tris-saline followed by 1:10 ConA-gold. My } incubations have been around 1 hr at room temp. Nary a gold particle do I } see under any of the conditions mentioned. Apparently, using fluorescently } labelled ConA these little buggers light up like the Hale-Bopp comet, so I } am at a lost. Can anybody out there give me some suggestions. I also have } some material fixed in 4% PF alone, embedded in LRW that I have not done } anything yet with. } Thanks in advance, } } Hank Adams } Electron Microscopy Laboratory } New Mexico State University } Las Cruces, NM 88003 } } http://www.nmsu.edu/Research/artsci/public_html/eml/ } } 505-6463600 } Since the glycocalyx is on the outside of the bacteria and you don't have to worry about sectioning to expose the conA to the glycocalyx, why don't you stain with con A-Au, wash thoroughly (centrifugations), and then embed normally?
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} Date: 22 Mar 1997 18:39:10 -0500 } From: Paul Webster {paul.webster-at-Yale.edu} } To: "Microscopy -at-MSA.Microscopy.Com" {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Re: Lectin Staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hank Adams writes: } "Hello, I am having no luck attempting to stain the glycocalyx? of a } species of sulfate-reducing bacteria using ConA-conjugated gold (5nm)." } (trimmed). } } Dear Hank, } Your problem may lie in the ConA-conjugated gold. Often, these probes very } seldom store well. If most of the protein has dissociated from the gold } particles, you will have a large amount of very specific ConA binding but no } gold to show you it is there. It is easy to test at the LM level by probing } sections treated with ConA-god with anti-ConA and fluorescent antibody. } } Once again, the best visualization method for lectins (re:ricin) may be to use } antibodies instead of the directly conjugated gold probes. If you prefer to use } ConA-gold then I recommend that you make your own. A slightly less satisfactory } approach would be for you to centrifuge down your ConA-gold (be careful not to } spin it so hard that is aggregates into the bottom of the tube) and resuspend it } in fresh PBS. This will remove most of the free protein. } } As an aside and in reference to a previous posting, streptavidin-gold also } suffers from this storage problem. When using the probe at the LM level, with } silver enhancement, or with multiple antibody layers the loss of labeling } efficiency is not so noticeable. However, attempting to visualize biotinylated } antibodies directly with streptavidin-gold will only work well for the first } week after the probe has been made. For situations where there are large } numbers of bound antibody, this effect may not be noticeable, but for situations } where there are only low numbers of antigens, it is very obvious. } } Making colloidal gold and coupling it to proteins or antibodies is easy and } anyone with even limited experience can do it. } } Check out my WWW "gold page" to see how easily it can be done. Be warned, if } you want to do this you usually need large supplies of protein (a point anyone } coupling antibodies should be aware of). } } Paul Webster } Center for Cell Imaging } Yale School of Medicine } http://info.med.yale.edu/cellimg } Also, if you're going to spin out your con A-Au from the uncongugated con A, keep the pH high (probably above 8-8.2, or whatever pH the original stuff was shipped in). Otherwise, you might have one big blob at the bottom of your tube. When we suspect unconjugated protein, we dilute the antibody-Au to the concentration we want to use with buffer at a pH above the pI (pH } 8), and then spin in a Beckman Airfuge 5 min at 30 lb (~100,000 g), discard the sup and resuspend the pellet in the same vol(pH 7) as we started with. Haven't tried con A; I assume it might work the same way???
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} Date: Sat, 22 Mar 1997 20:46:17 -0800 } From: Eric {earosen-at-pop.goodnet.com} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Lectin staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } You may also want to treat the sections with a very low concentration } of sodium periodate which will destroy the section if you are not } careful.. This will expose some of the bnding sites fo the Con-A.. or } on the other hand the Lctins are too old, and the gold has fallen off } of the lectin which occured to me also when I was using gold labeled } lectins that were stored in the fridge........ So I got very } scattered labeling or no labelling at all but when used in teh LM } with Cy3 they light up like a chrsitmas tree.... } } lAlso you may want to try a lower comcentration of fixatives in } Formaldehyde and Glut. } } } --------------------------------------------------------------------- } --- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Hello, I am having no luck attempting to stain the glycocalyx? of a } species of sulfate-reducing bacteria using ConA-conjugated gold (5nm). } Using 4%PF+0.1%Glute in Cac buffer fixation followed by LRW embedding } I have tried 1:10 to 1:100 in P04 buffer pH 7.2. I have also tried } ConA-gold in Tris-saline with and without 1mM CaCl2 and 1mM MgCl2 at } pH 6.9,7.2 and 8.0. diluted 1:10-1:100. Next I tried taking a fresh } culture and washing several times in Tris-saline followed by 1:10 } ConA-gold. My incubations have been around 1 hr at room temp. Nary a } gold particle do I see under any of the conditions mentioned. } Apparently, using fluorescently labelled ConA these little buggers } light up like the Hale-Bopp comet, so I am at a lost. Can anybody out } there give me some suggestions. I also have some material fixed in 4% } PF alone, embedded in LRW that I have not done anything yet with. } Thanks in advance, } } Hank Adams } Electron Microscopy Laboratory } New Mexico State University } Las Cruces, NM 88003 } } http://www.nmsu.edu/Research/artsci/public_html/eml/ } } 505-6463600 } } } } \\|// } (o o) } ~~~~~~~~~~~~oOOo~(_)~oOOo~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } { { {This message is made of 100% recycled electrons} } } } } Cheers ;o) :o) %o) } Eric {Mesa Arizona} } http://www.goodnet.com/~earosen (Note the tilde before earosen) }
Etching agents are not usually required for porous acrylics.
If you want to test your fixation, try various ones and then fluorescent labeling--saves a lot of work doing it at the EM level.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
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Reply to: RE} Sun's Question
What you saw probably were bend extinction contour. (See Hersch et al's book "Electron microscopy of thin crystals", page 203) The change of the pattern may be caused by beam heating.
-------------------------------------- You wrote:
I am a student now using Hitachi-8100 TEM to study submicrometer Cu-Zn alloy particals. The particals are mostly six-fold symmetry plates. When the beam focused on a partical at a high magnification, some dark stripes appear in the partical's bright-field image and change their shapes until a snow-flake-like pattern is formed. The pattern comprised of six center-crossed dark stripes. Select-area diffracton shows a single crystal diffracton pattern with six-fold symmetry. But dark-field image shows that one single diff spot is not coming from the whole partical but from ONE corresponding dark stripe.
My questions are: 1)Do anyone out there ever find similar phenomenon? 2)How can I know the temperature of the particals under the brightening of the electron beam? I think temperature is a reason for the formation of the snow-flake-like pattern.
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Subject: Time:4:09 PM OFFICE MEMO Position Available at NCEM Date:3/23/97
Staff Materials Scientist
Materials Sciences Division National Center for Electron Microscopy (NCEM) Job MSD/5001
Essential -- The NCEM is a national user facility with several state-of-the-art electron microscopes and advanced image analysis and specimen preparation facilities, dedicated to the electron-optical microcharacterization of materials. The Center has an opening for a Staff Scientist to lead its research effort in high-resolution imaging of materials. Conduct original research into the development of high-resolution techniques and their application to significant materials problems. Responsible for the operation and development of several high- resolution microscopes, including a new 300kV field emission instrument and the 1MeV ARM. Lead a program to extend instrument resolution toward the 1A information limit.
As an essential member of a national user facility, the candidate will have the opportunity to collaborate with a broad range of investigators from the national and international scientific community. The position also involves close collaboration with other staff members on the development of computer analysis, and control and supervision of postdoctoral and technical staff.
The candidate will be able to contribute substantially to the successful operation and future development of the facility.
QUALIFICATIONS: Essential -- Outstanding track record in development and application of quantitative high-resolution transmission electron microscopy. Extensive practical experience with advanced high-resolution imaging and analysis techniques, such as series reconstruction, holography, and image interpretation. Demonstrated ability to initiate collaborations and carry out high- quality research, using the unique facilities of the NCEM. Marginal -- Ph.D. in physical sciences.
POSTING DATE: March 6, 1997. CLOSING DATE: Open until filled.
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Your labelling problem may simply be that you are using resin sections which are usually not permeable to gold labels (ie you only present a very small edge of the bacteria in e.m. whereas I assume the light microscopy that you mention was done on whole cells which present a much larger surface to the fluorescent label. One test might be to try and use the fluorescent label on LR White sections and examine that under the fluorescent light microscope.
Malcolm Haswell University of Sunderland UK ----------
Dear Sun, As someone else stated, the dark lines you saw in bright field are bend contours. You saw three crossed bend contours, each which corresponds to a particular diffraction condition. Thus where all three bend contours cross, all three diffraction conditions are operating simultaneously and you are looking down the zone axis of the crystal. When you work in dark field you will be imaging with only one of the diffraction spots so you will only see one bend contour (which will now be bright). As for why the pattern changes on observation, this is quite common and you will find that many things deform or tilt slightly under the heat of the beam. I don't know about your system but in some systems, the heating can induce crystallisation which would obviously change the contrast pattern. Lastly, I can't think of a simple way of measuring the temperature of your particles.
Yours sincerely
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
If you have full World Wide Web access you could try using a search engine such as Lycos or Yahoo (eg microscope, microscopy, amateur, hobby). It should turn up some useful web pages which could lead to further links.
One really excellent on-line organization & magazine is: Microscopy-uk http://www.microscopy-uk.org.uk/#top which has the online magazine 'Micscape'. It has had excellent articles on everything from hand lenses to electron microscopes and makes few assumptions that it's readership are pro microscopists. There are also lots of links.
Other places to try would be the home pages of the Microscopy Society of America (US) and Royal Microscopy Society (UK) and again look for useful links.
I hope this helps
Malcolm Haswell University of Sunderland UK
----------
Can any one tell me of a Microscope news group taylored toward the newbie into this hobby? Also, are there any good magazines on the market specializing in MIcroscopes and the Microscope hobby?
I'd like to thank Marks, Dorai, Garber, Yi Huang and Ian MacLaren for their suggestions to my questions.
Bend contours and multiply twins are possible reasons for the formation of the pattern described in my questions. I will find more details on these matters.
A commercial source for Aclar is ProPlastics, Inc, P.O. Box 679, Linden, NJ 07036 201-925-5555. They have a minimum order. Several years ago we bought two pounds of 5mil for $75.00. It will be a lifetime supply.
At 03:55 PM 3/21/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Hi Gary, Aclar is great! Read the paper in Journal of Electron Microscopy Technique 10:77-85 (1988), by Kingsley and Cole. I have used it in epon embedding protocols. There is nothing like it. You can get it from Electron Microscopy Sciences. Call them since it may not be listed in their catalogue. Ask to speak to Stacie Kirsch.
Aclar is a sturdy thin film, transparent as glass, no detectable autofluorescence, chemically inert, sections beautifully, and so on and so on.
Have fun.
Sally
On Fri, 21 Mar 1997, Gary Radice wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Several of you have recommending using Aclar film in your UV polymerization } protocols. What is Aclar film, and where do you purchase it? } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX) } } }
(I am posting this for a colleague in France who is not on this list. Contact him, not me, please.) ==============================
My laboratory has bought recently a SX-100, so I should be happy to find somebody who might be interested in purchasing our Camebax Electron Microprobe, which is in good condition.
Here is a short description : Camebax with 3 WDS and 1 EDS for fully compatible coupled analysis WDS-EDS (Quantitative analysis on fixed points or lines, X-ray mapping) WDS : all usual crystals, plus PC1 and PC2 for light elements EDS : Tracor model, with Be window Equiped with Back-Scattered Electron detector, and anticontamination system. Software : all the software provided by Tracor (Noran) for TN-5500 : - to control spectrometers, sample holder, and faraday cup - to perform quantitative analysis (ZAF and PRZ) - to make standardless analysis (EDS) - Image analysis (by digitalisation)
The column is from 1976, but everything else is from 1988 (including spectrometers)
Price = $30 000 USD. (shipping to be paid by purchaser)
For more information, contact:
Michel LAHAYE ICMCB, Universiti Bordeaux I, Pessac (FRANCE) Tel 33 5 56 84 62 93 Fax 33 5 56 84 27 61 e-mail : lahaye-at-icmcb.u-bordeaux.fr ==================================================================
Jussi: We could only find Swift point counters when we were in the market for a new counter for mineralogical applications. They were still in business 5 years ago when we bought our newer electronic model. There may be other companies selling similar products. We had an ancient one (} 35 years)that with maintenance the stage worked, but the old electric pushbuttons were in bad shape. It is dependable and a very adaptable system. If you get all the gear sets, you can count at any optimum step size typical of optical microscope use. We used it primarily for opague ore minerals in polished briquettes. The U.S. office is in San Jose CA. Fax 408-292-7967. I'm sure there is a closer one to you in Europe, but I don't have any info on it. You might try looking on the WWW for a webpage.
I am presently unemployed and have no financial interest in Swift or anything at present, but would be glad to have some! ================================================ Michael L. Boucher Sr. mboucher-at-isd.net 13345 Foliage Avenue Apple Valley, MN 55124-5603 Ph 612-432-8836 ================================================
On Thu, 20 Mar 1997, Ginger Baker wrote: } } Hello all, } } I have a PhD student from Brazil who is studying Catolaccus } grandis. He places a female on top of a beeswax cell which contains } boll weevil larva. The female deposits an egg on the larva and 2 } weeks later, it matures and burrows its way out of the beeswax } creating a hole. He would like to study and measure these holes. } } However, from past experience I know that beeswax melts when placed } in my Au/Pd sputter coater. Is there any way to coat the beeswax and } examine them with an SEM? Or should he try another route? } } Thank you in advance,
} Email: lizard-at-okway.okstate.edu } Hi Ginger, I guess the coater you used is a diode type of sputter coater. Heat is generated during coating. In order to solve your problem, you should to use a kind of "cooled sputter coater"-- planar magnetron sputter (PMS) coater. A permanent magnet is posstioned at the center of the cathode to deflect the electrons away from the specimen.
On Thu, 20 Mar 1997, Ginger Baker wrote: } } Hello all, } } I have a PhD student from Brazil who is studying Catolaccus } grandis. He places a female on top of a beeswax cell which contains } boll weevil larva. The female deposits an egg on the larva and 2 } weeks later, it matures and burrows its way out of the beeswax } creating a hole. He would like to study and measure these holes. } } However, from past experience I know that beeswax melts when placed } in my Au/Pd sputter coater. Is there any way to coat the beeswax and } examine them with an SEM? Or should he try another route? } } Thank you in advance,
} Email: lizard-at-okway.okstate.edu } Hi Ginger, I guess the coater you used is a diode type of sputter coater. Heat is generated during coating. In order to solve your problem, you should to use a kind of "cooled sputter coater"-- planar magnetron sputter (PMS) coater. A permanent magnet is positioned at the center of the cathode to deflect the electrons away from the specimen.
Ya Chen
Ya Chen
========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
I am looking for a source of adapters to convert the old short barrel Leitz objectives to the newer long barrel type (170mm tube length).
I also need a set of PLAN APO's for a Zeiss Photomicroscope in 160mm tube length. Need 4X, 10X, 20X, 40X and 100X O.I. or anything close in Plan Apo's.
Also need the PHACO 2 and PHACO 3 inserts for a Leitz Laborlux-11 (type 55 condenser).
Any leads would be most appreciated.
Cordially, Gil Groehn (313) 884-1139
-- ============================================================= ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org Grosse Pointe Farms, MICH 48236 USA PHONE: 313-884-1139 ===============================================================
Reprosil is good, and so are any of the other polyvinyl siloxane impression materials, available from any dental supplier. The resin can be Epon, or 302-1 from Epo-Tek or anything from the EM world.
Lesley Weston
On Fri, 21 Mar 1997, Ronald LHerault wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } An alternative may be replication using Reprosil, a Polyvinyl Siloxane } from Caulk. It makes a negative which can be transfered to a positive by } using Spurr low viscosity embedding medium. Detail is there albeit a } little softened. } } Ron } }
I was looking at your page and hoped you might help me. I am looking for a Windows based SEM capture board. We do quite a bit of optical microscope image capture, but nothing yet on SEMs.
I have heard there are a couple out there, but haven't found them yet!
Good afternoon, I was wondering if those of you who successfully make your own holey grids for stigmation would care to share the method. I've been using glcerol and 0.25% Formvar (old bottle) and while I can produce small, nice looking "holes", on close inspection they're film. There are some true holes, but few and far between. Any suggestions? Many thanks in advance.
Dwight Beebe E-mail: beebed-at-ere.umontreal.ca Institut de recherche en biologie vegetale Voice: 514-872-4563 Universite de Montreal FAX: 514-872-9406 4101, rue Sherbrooke est Montreal, Quebec H1X 2B2 Canada
We historically have made our own holey films, because we could not buy good holey support films from any manufacturer. Without question, it is an art, and most of the time a major pain in a posterior region. I would be happy to send you the detailed instructions we have found to be most reproducible.
Recently, however, we purchased some holey films from SPI, which have been uniformly *gorgeous*. They are thin, clean, a large fraction of holes etc. Probably equivalent to the best I have ever made. I don't recall the price, but my impression is that I can't make them as cheaply, so why bother.
Highly recommended.
Larry
PS I have no particular connection to SPI.
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Dear Dwight, I know of two methods of preparing holey films. The first is to shake up a mixture of liquid soap and water and Collodion until it froths, then drop a drop or two on the surface of distilled water. Lay the grids on top of the Collodion layer, pick up with a filter paper, dry and carbon coat. Put the grids in a Jaffe washer with cloroform for 48 hours to remove the Collodion. The other method, if you have Nucleopore type filters, is to carbon coat a strip of Nucleopore filter film, place a square of the coated filter on the grid sitting on a nickel mesh on the surface of the Jaffe washer full of cloroform, wait 48 hours to dissolve the Nucleopore, dry grids. The holes in the Nucleopore will be now be holes in the carbon coat. I must admit I have not tried the first method myself, but someone in my lab did, years ago. You wrote: } I was wondering if those of you who successfully make your own } holey grids for stigmation would care to share the method. I've been using } glcerol and 0.25% Formvar (old bottle) and while I can produce small, nice } looking "holes", on close inspection they're film. There are some true } holes, but few and far between. Any suggestions? Many thanks in advance. } } } Dwight Beebe E-mail: beebed-at-ere.umontreal.ca } Institut de recherche en biologie vegetale Voice: 514-872-4563 } Universite de Montreal FAX: 514-872-9406 } 4101, rue Sherbrooke est } Montreal, Quebec H1X 2B2 } Canada Good luck, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I am looking for any information on the fluorescent dye: Commercial name; 'Geranine G', C.I. name; Direct Red 48, C.I. number; 14930. I know that it is a monoazo dye with good affinity for certain proteins, so my question is does anyone have any further information on Geranine G, such as Excitation and Emission wavelengths, or even manufacturers or distributors that carry this dye. Failing this would anyone know of a substitute of similar properties.
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Ron Herault wrote: ----------------------------------------- An alternative may be replication using Reprosil, a Polyvinyl Siloxane from Caulk. It makes a negative which can be transfered to a positive by using Spurr low viscosity embedding medium. Detail is there albeit a little softened. ----------------------------------------- Another alternative, possibly an even better choice, would be our own "Wet Surface Replica Kit", it too is silicone based, but it has been more optimized for SEM applications. It can be cured into a "negative" much more quickly, and when converted, the "positive" replicating system is much easier to use than the "Spurr" approach and on a "per replica basis" is a lot lower in cost. A demo set of micrographs made from such positive replicas, on human skin, can be seen in our electronic catalog at the website mentioned below.
Disclaimer: SPI Supplies manufactures the above mentioned "Wet Replica Kit" and would have a vested interest in seeing more persons using it.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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In message {199703241633_MC2-1336-770C-at-compuserve.com} , "Seth J. Grotelueschen" {sethg-at-CompuServe.COM} writes
Dear Seth
Is is not obvious which country you are from but if you are interested in someone in the UK try Deben Research who will be able to help you. If you want the full address and contact no let me know.
They also support our imaging software.
Best regards
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We are studying some ultrastructural features of a new type of epidermal trichomes on plants. Trying to define and quantify ulstrastructural changes during the cell development (from meristematic stage to fully differentiated trichome) we have to deal with the following questions. How to start analysing TEM micrographs in order to reach general conclusions on variations of organelle numbers ? Are there easy methods to start quantitative morphological analysis ? Do we have to work with image analysing softwares or not ? How do you deal with this kind of problem ? Do you know about books or papers on similar studies that can be used as references?
Any solutions are welcome... Thanks !
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
ULTRAMICROTOMY ANY CRYO METHODS FOR MATERIALS SCIENCE
PLACE: UNIVERSITY OF PENNSYLVANIA LABORATORY FOR THE RESEARCH ON THE STRUCTURE OF MATTER PHILADELPHIA, PA 19104 DATE: JUNE 11-13,1997 SPONSORS: LEICA INC., DIATOME U.S., ELECTRON MICROSCOPY SCIENCES
COURSE SPEAKERS AND INSTRUCTORS: Dr. Tom Malis, Phil Swab, Helmut Gnagi
OVERVIEW:
This workshop will cover the use of Room Temperature and CryoUltramicrotomy techniques as it applies to Material Science. Included in the course will be all of the preparation leading up to sectioning, such as, embedding, trimming and staining on different types of industrial specimens(polymers, metals, plastics, fibers, rubbers, powders, foils, etc.). From all of the topics to be covered and in depth review of all of the different techniques will take place and there will be an emphasis on "Hands-On" lab time. Participants are encouraged to bring their own samples as well. Our goal, upon completion of the course, is that each of the participants will be able to return to there own lab with a better understanding of the theory and principles behind ultramicrotomy, and the ability to successfully perform all of the techniques which we shall cover.
COURSE COST: $1,500.00 The price includes: Hotel accommodations(3 nights), lunch daily, continental breakfast daily, 1 group dinner, all course supplies and full lab time.
FIRST COURSE DEADLINE: April 18, 1997
Parties that are interested may either respond by E-Mail to SGKCCK-at-AOL.COM or Leica's seminar voice mail to 1-800-248-0665 EXT:5010 Contacts are Stacie Kirsch or Ann Korsen. To confirm your space in the course below is a form that may be filled out and either E-Mailed to SGKCCK-at-aol.com or faxed to 215-646-8931 ATT: Stacie Kirsch. All checks should be mailed to Diatome U.S. P.O. Box 125, Fort Washington, Pa 19034 and made out to Leica, Inc.
WORKSHOP APPLICATION FORM: Company Name:________________________ Participant's Nmae:______________________ Full Address:____________________________ City, State, Zip:__________________________ Tel #:___________________________________ Fax #:___________________________________ E-Mail:__________________________________ Best Time To Reach:______________________ Area of Interest: Cryo or Room Temperature Type of Material Working With:______________ Will you be bringing your own sample?___________________What:________
Paying by PO#:________________________(Please mail to P.O. Box) Check#:_______________________________
If paying by Purchase order number please be sure to include your full billing address.
The normal replicas of ruled gratings allow SEM magnification calibration up to about 50 000x mag. At magnifications higher than this it is not so easy to find good accurate SEM calibration specimens.
What SEM calibration specimens are available for use at 50 000 to 1 000 000 x mag in an In-lens FEGSEM?
Regards,
Dr Jan Coetzee Head: Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria 0002 Internet:janc-at-ccnet.up.ac.za South Africa http://www.up.ac.za/academic/electron/emunit1.htm
Hello, A recent upgrade in EDS instrumentation and detectors has given us the ability to detect low elements (C, N, O). I have been detecting a peak that corresponds to nitrogen in all the titanium spectra that I have acquired. This has occurred using pure titanium (T18) or alloyed titanium (beta ti). The nitrogen peak ranges from 5-10 weight percent of the spectra. I know that certain elements can implant in Ti, including C and N, but these samples have not undergone the heating conditions that would favor ion implantation. The N peaks do not increase under long e-beam bombardment. The spectra are collected under routine conditions (15KeV, recommended WD, 25% deadtime, 100 sec, standardless analysis but the calibration seems correct). I would appreciate it if anyone could shed some light on this problem.
Wallace Ambrose Dental Research Center Univ. of North Carolina Chapel Hill, NC
The 9th ed of Conn's Biological Stains offers little on Geranine G, only a few references. Try the Biological Stain Commission, Dept. of Pathology, Univ. of Rochester Medical Center, Rochester, NY (716)-275-2751.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Ambrose, Wallace wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, } A recent upgrade in EDS instrumentation and detectors has given us the } ability to detect low elements (C, N, O). I have been detecting a peak } that corresponds to nitrogen in all the titanium spectra that I have } acquired. This has occurred using pure titanium (T18) or alloyed titanium } (beta ti). The nitrogen peak ranges from 5-10 weight percent of the } spectra. I know that certain elements can implant in Ti, including C and } N, but these samples have not undergone the heating conditions that would } favor ion implantation. The N peaks do not increase under long e-beam } bombardment. The spectra are collected under routine conditions (15KeV, } recommended WD, 25% deadtime, 100 sec, standardless analysis but the } calibration seems correct). I would appreciate it if anyone could shed } some light on this problem. } } Wallace Ambrose } Dental Research Center } Univ. of North Carolina } Chapel Hill, NC
Just a quick thought: are you sure that it is a nitrogen peak and not Ti L, or Ti L Escape peak? --
Naresh Shah Associate Research Professor, University of Kentucky Department of Chemical and Materials Engineering (CME) Consortium for Fossil Fuel Liquefaction Science (CFFLS) 533 South Limestone Street, Room 111 Lexington, KY 40508-4005 Phone: (606) 257-5119; FAX: (606) 257-7215; e-mail: naresh-at-pop.uky.edu
} A recent upgrade in EDS instrumentation and detectors has given us the } ability to detect low elements (C, N, O). I have been detecting a peak } that corresponds to nitrogen in all the titanium spectra that I have } acquired.
This is the Ti L-l line (the transition of the M-I to the L-III subshell), a throroughly nasty interference, even in WDS. Contact me if you have problems with it.
Alfred
----------------------------------------- Alfred Kracher akracher-at-iastate.edu http://www.public.iastate.edu/~akracher -----------------------------------------
If your SEM screen accepts a Polaroid camera, Polaroid makes a "MicroCam" and "PhotoPad" color scanner. Together they retail for about $1000. I have only seen the ad, and I have no first hand information (Polaroid: 1-800-662-8337). I have no connection to or investment in Polaroid.
There are other manufacturers of frame-grabbers and videoboards for SEMs, but alas, I discarded the information.
Scott Schwinge Friday Harbor Labs University of Washington
} Hi, } } I was looking at your page and hoped you might help me. } I am looking for a Windows based SEM capture board. We } do quite a bit of optical microscope image capture, but } nothing yet on SEMs. } } I have heard there are a couple out there, but haven't } found them yet! } } Thanks } Seth Grotelueschen
Can anyone suggest names of service engineers with experience on vintage Cambridge electron microscopes? I have a 1971 Cambridge Stereoscan S4 SEM in need of service. I'd like to locate an experienced technician in the metro Atlanta area or the southeastern US, but I would appreciate all references and information. Thanks,
Michael Knotts ------------------------------------------------------------- Michael E. Knotts, Ph.D. E-mail: ph281mk-at-prism.gatech.edu Contributing Editor {The Light Touch} OPTICS & PHOTONICS NEWS Georgia Tech / School of Physics / Atlanta, GA 30332-0430 Tel: (404) 894-3422 FAX: (404) 894-9958
Although titanium readily attracts nitrogen and oxygen, I think your problem is related to peak overlap.
A quick look at x-ray tables reveals:
El/line keV
N Ka1&2 0.392
Ti Ll 0.395
Ti La1&2 0.452
O Ka1&2 0.523
The N Ka and Ti Ll peaks cannot be resolved by typical EDS spectrometers, since you would need a resolution of ~2 ev. To resovle the N Ka and Ti La peaks you would need a detector resolution of ~50 ev.
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electron-sample interaction programs Hello, I am in need of a Monte Carlo based program (PC or Mac) that models the interaction of an electron beam with a user defined sample. We have a 200 keV TEM and an SEM capable of imaging at low voltage; hence, the program should handle electron transparent samples up to bulk materials and a ~1 to 200 keV electron beam of variable probe size. I am not only interested in the electron interaction volume but also in the volume from which many other signals can be collected, e.g., backscattered electrons, x-rays, secondary electrons, Auger electrons, etc.
I have seen electron flight simulator for sale ($500), but have little info on what it provides. What about quality shareware?
Thank you
Brad Storey Materials Scientist Argonne National Lab 208-533-7685
} A recent upgrade in EDS instrumentation and detectors has given us the } ability to detect low elements (C, N, O). I have been detecting a peak } that corresponds to nitrogen in all the titanium spectra that I have } acquired. This has occurred using pure titanium (T18) or alloyed titanium } (beta ti). The nitrogen peak ranges from 5-10 weight percent of the
This is not a nitrogen peak. You are seeing the Ti L-series lines, which exhibit a strong overlap with the Nitrogen K-series lines.
This overlap is bad enough that it cannot really be easily resolved even using a wavelength-dispersive spectrometer. It is one of the classic overlaps that makes analysis of Ti alloys for nitrogen problematic.
Nitrogen detection is hampered by strong absorption of Ni Ka by any carbon that is in the x-ray path, be it in the sample, the carbon coat on the specimen, oil on the EDS detector window, window material (like diamond, for example), etc.
These factors make nitrogen measurement a challenge, since one does not expect to find it in any real concentration unless it is a distinct phase (like TiN).
Paul
+----------------------------------------------------+ | Paul K. Carpenter paulc-at-gps.caltech.edu | | Division Analytical Facility | | Geological and Planetary Sciences MC 100-23 | | California Institute of Technology | | Pasadena, CA 91125 | | 818-395-6126 (X-ray Lab) 818-568-0935 (Dept. FAX) | +----------------------------------------------------+
I use a method by Kuga and Brown from Philips Electron Optics Bulletin v126, p.19 (1989) to make lacy formvar grids. First a note of caution. I use dichloroethane as my solvent for the formvar resin. After having much difficulty in preparing my films, I was told that when exposed to light, dichloroethane slowly forms HCl in the solution. The HCl destroys the integrity of the formvar polymer. Solution: store the formvar solution in a brown bottle in a dark cabinet.
Method:
- I use 0.25% formvar solution in dichloroethane and freshly cleaved mica sheets as a substrate. The formvar lifts off the mica much easier than from glass microscope slides.
- Dip the mica into the formvar solution, wick off excess on a paper towel, then breathe heavily on the mica for about 5 seconds while it is still wet. The moisture in your breath condenses in the solution. Caution: Don't inhale!
- When the mica has dried, score the edges and float off on water.
- Place 200 mesh TEM grids on the floating film. The area with the best holes will appear milky.
- I then take saran wrap, stretch it tightly across the mouth of a small (~100ml beaker), and press it down at a slight angle onto the floating formvar film. The film will stick to saran wrap and you can easily pick it up off the surface of the water.
- After the film has mostly dried, pick up the grids from the saran wrap "drumhead" and place them on filter paper.
- The film will have many "pseudoholes" that have a thin residual film across them.
- Following the method of Kuga and Brown, by heating the film to about the T(g) temperature you can break these holes open. The time and temperature are critical. I place the bare filter paper in a small lab oven (not in a petri dish; it has too much thermal mass) at 110C for 12 minutes.
- I then coat both sides with carbon to stabilize the formvar lace.
I can easily make 50-100 grids in an hour (exclusive of the carbon coating). These grids make wonderful supports for looking at fine particulate dispersions.
Cheers, Henk
} } Good afternoon, } I was wondering if those of you who successfully make your own } holey grids for stigmation would care to share the method. I've been using } glcerol and 0.25% Formvar (old bottle) and while I can produce small, nice } looking "holes", on close inspection they're film. There are some true } holes, but few and far between. Any suggestions? Many thanks in advance. } } } Dwight Beebe E-mail: beebed-at-ere.umontreal.ca } Institut de recherche en biologie vegetale Voice: 514-872-4563 } Universite de Montreal FAX: 514-872-9406 } 4101, rue Sherbrooke est } Montreal, Quebec H1X 2B2 } Canada
Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility ------------------------------------------------------------------- Prosperity is the blessing of the Old Testament; Adversity is the blessing of the New. Francis Bacon, "Of Adversity."
Does anyone know of a source for teflon o-rings with a thickness of {100 uM?
Thanks, Sandy Simon
Sanford M. Simon Laboratory of Cellular Biophysics Box 304 Rockefeller University 1230 York Avenue New York, N.Y. 10021 212-327-8130 (voice) 212-327-8022 (fax) simon-at-rockvax.rockefeller.edu (e-mail)
Does anyone have a current phone number for Energy Beam Sciences or whichever company has taken over Polaron? I have a question about our Polaron film thickness monitor. Thanks for your help.
} Dear All, } } The normal replicas of ruled gratings allow SEM magnification } calibration up to about 50 000x mag. At magnifications higher than } this it is not so easy to find good accurate SEM calibration } specimens. } } What SEM calibration specimens are available for use at } 50 000 to 1 000 000 x mag in an In-lens FEGSEM? } } Regards, } } } } Dr Jan Coetzee
In biological TEM, colloidal gold in a variety of immunological labeling techniques. This gold has fairly well defined size distributions - the smallest available has a mean size of 1nm. If you check the EM supplies catalogues (Agar, SPI, Ted Pella, etc), you should find it - personally, I've never tried it, but I don't see any reason why it couldn't be used (although you might need to remove some antibodies).
The other approach is to use the same techniques that is used in TEM, where ther is a bit of a gap between gratings and lattice resolution specimens. Basically, you work up step by step, going from a lower mag, where the grating gives you a calibration, identify a number of obvious features, then go up in mag and relocate the same features - they got to be the same distance apart. A bit tedious and not statistically very accurate, +/- 5% at best, but its better than nothing.
Brad Storey wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } electron-sample interaction programs } Hello, } I am in need of a Monte Carlo based program (PC or Mac) that models the } interaction of an electron beam with a user defined sample. We have a 200 keV } TEM and an SEM capable of imaging at low voltage; hence, the program should } handle electron transparent samples up to bulk materials and a ~1 to 200 keV } electron beam of variable probe size. I am not only interested in the electron } interaction volume but also in the volume from which many other signals can be } collected, e.g., backscattered electrons, x-rays, secondary electrons, Auger } electrons, etc. } } I have seen electron flight simulator for sale ($500), but have little info on } what it provides. What about quality shareware? } } Thank you } } Brad Storey } Materials Scientist } Argonne National Lab } 208-533-7685
On my laboratory web site are informations about almost all public domain and commercial Monte Carlo programs for PC and Mac's.
} Hello, } A recent upgrade in EDS instrumentation and detectors has given us the } ability to detect low elements (C, N, O). I have been detecting a peak } that corresponds to nitrogen in all the titanium spectra that I have
snips
} calibration seems correct). I would appreciate it if anyone could shed } some light on this problem. } } Wallace Ambrose } Dental Research Center } Univ. of North Carolina } Chapel Hill, NC
Can anyone refer me to a basic reference on electron channeling = contrast, or failing that, give me a few hints on optimal conditions for = observing this type of contrast ?=20
TIA=20
Glenn
Glenn Poirier Phone (514) 398 6774 Electron Microprobe Laboratory Fax (514) 398 4680 Earth and planetary Science=20 Mcgill University
THERE ARE THREE SIDES TO EVERY STORY: yOURS, MINE AND THE TRUTH
Talking of peak overlaps, is anyone out there capable of reliable accurate analysis of V in the presence of large amounts of Ti, using:
a WDS
b EDS?
I ask because I concluded recently that I couldn't, with my EDS-only system, analyse for V in titanomagnetites, and that it probably wasn't possible even with WDS. If I ask my system to look for V in my standard Rutile (TiO2), it always finds a few tenths of a %, even though my software (LINK ZAF4/FLS) strips out Ti Kb along with the Ka. I have no way of telling whether this V is genuine or not, but I suspect that it isn't, as my supposedly-pure Ti metal standard also gets credited with a small amount of V, which I doubt. I was recently given another rutile standard, and, blow me down, it is reputed to have 0.4% V by WDS analysis. Anyone got any feeling for whether this is likely to be true?
cheers
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Wallace: This is most likely the Ti Ll line overlapping N Ka. Beware of overlaps in the light element energy region!
Jim *.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.* James J. McGee (jmcgee-at-sc.edu) Dept. of Geological Sciences University of South Carolina (803) 777-6300 (Office) Columbia, SC 29208 (803) 777-6610 (Fax)
----------
Hello, A recent upgrade in EDS instrumentation and detectors has given us the ability to detect low elements (C, N, O). I have been detecting a peak that corresponds to nitrogen in all the titanium spectra that I have acquired. This has occurred using pure titanium (T18) or alloyed titanium (beta ti). The nitrogen peak ranges from 5-10 weight percent of the spectra. I know that certain elements can implant in Ti, including C and N, but these samples have not undergone the heating conditions that would favor ion implantation. The N peaks do not increase under long e-beam bombardment. The spectra are collected under routine conditions (15KeV, recommended WD, 25% deadtime, 100 sec, standardless analysis but the calibration seems correct). I would appreciate it if anyone could shed some light on this problem.
Wallace Ambrose Dental Research Center Univ. of North Carolina Chapel Hill, NC
The question of how to make holey carbon films came up on the list server two years ago. At that time John Gabrovsek (gabrovj-at-ccsmtp.ccf.org) gave the following reference: Baumeister & Seredynsky, Micron 1976, Vol 7, p. 49, and Jane Fagerland (fagerland.jane-at-igate.abbott.com)gave this one: Elsner, Proceedings, 29th EMSA Meeting, p. 460. Both methods were said to work satisfactorily. You might try contacting these people for more details.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Dear pascal veys: Here are some basic references that may help you get started: Weibel, E.R. 1979. Stereological methods, vol 1. Practical Methods for biological morphometry. Academic Press, New York.
Weibel, E.R., "Stereological Techniques for Electron Microscopic Morphometry". In Principles and Techniques of Electron Microscopy, Vol. 3, M.A. Hayat, ed. 1973. pp 239-296.
Loud, A.V., 1968. A quantitative stereological description of the ultrastructure of normal rat liver parenchymal cells. Journal of Cell Biology, Vol 37:27-46.
Don Gantz Boston Univ Med School gantz-at-med-biophd,bu.edu
Hi Seth, JEOL in Scandinavia market a system called Semafore, their Web page may be worth a look as it has sample software to download. email me if you would like more information.
all the best Pete Lander (JEOL UK)
} -----------------------. } } } } Hi, } } } } I was looking at your page and hoped you might help me. I am looking for a } } Windows based SEM capture board. We do quite a bit of optical microscope } } image capture, but nothing yet on SEMs. } } } } I have heard there are a couple out there, but haven't found them yet! } } } } Thanks } } Seth Grotelueschen }
Return messages: Home Work email pal-at-fenland.demon.co.uk pal-at-jeolsys.demon.co.uk phone 01354 661413 01707 377117 fax phone first 01707 373255
The MicroCam slides onto the eyepiece of a optical microscope. It doesn't work with a SEM camera port. John D. Warren Area Sales Manager Digital Products Polaroid Corporation
4525 Leonard Parkway Richmond, Virginia 23221-1809
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If your SEM screen accepts a Polaroid camera, Polaroid makes a "MicroCam" and "PhotoPad" color scanner. Together they retail for about $1000. I have only seen the ad, and I have no first hand information (Polaroid: 1-800-662-8337). I have no connection to or investment in Polaroid.
There are other manufacturers of frame-grabbers and videoboards for SEMs, but alas, I discarded the information.
Scott Schwinge Friday Harbor Labs University of Washington
} Hi, } } I was looking at your page and hoped you might help me. } I am looking for a Windows based SEM capture board. We } do quite a bit of optical microscope image capture, but } nothing yet on SEMs. } } I have heard there are a couple out there, but haven't } found them yet! } } Thanks } Seth Grotelueschen
Don't feel alone. Last year, I tried to do a quantitative analysis of 6/4 titanium/vanadium by EDS. The responses you have received so far explain the problem - peak overlaps. I had the additional factor of a V L-alpha emission at 0.51 keV. I opted for emission spectroscopy at an outside lab.
Regards,
-Bob **************************** Bob Citron Chiron Vision Claremont, CA Bob_Citron-at-cc.chiron.com ****************************
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Hello, A recent upgrade in EDS instrumentation and detectors has given us the ability to detect low elements (C, N, O). I have been detecting a peak that corresponds to nitrogen in all the titanium spectra that I have acquired. This has occurred using pure titanium (T18) or alloyed titanium (beta ti). The nitrogen peak ranges from 5-10 weight percent of the spectra. I know that certain elements can implant in Ti, including C and N, but these samples have not undergone the heating conditions that would favor ion implantation. The N peaks do not increase under long e-beam bombardment. The spectra are collected under routine conditions (15KeV, recommended WD, 25% deadtime, 100 sec, standardless analysis but the calibration seems correct). I would appreciate it if anyone could shed some light on this problem.
Wallace Ambrose Dental Research Center Univ. of North Carolina Chapel Hill, NC
Does anyone have a current phone number for Energy Beam Sciences or whichever company has taken over Polaron? I have a question about our Polaron film thickness monitor. Thanks for your help.
Hi there: My name is Miguel Sanchez my problem is the following As part of my business I receive micron rated textiles, that is nylon mesh that has a determined distance between threads. Sometimes is very difficult to determine that the textile is inside tolerances or not. I use an optical microscope to do measure them my question is: What do I need to measure the textiles with the use of a camera and a computer? I want to use a camera so we can agree on what we are measuring. The solution has to be cost effective. Very cost effective. Your comments will be highly appreciated Thanks
while studing ZrO2 thin film on sapphire substrate, I found a kind of modulated structure like a wave near the interface region (It's not a morie fringe). This kind of structure is very similia to the pre-martensitic sructure found in Ni-Al system. The lattice spacing in the modulated region is the same as (111) d-spacing, while in the uper region d=d(002). The film has a (001)//(0001) prefer orientation with sapphire. What I want to know is: 1. How the modulated structure formed, and how (111)//(0001) orientation can change to (001)//(0001)? Can the modulated region comes from a martensitic transformation? 2. Is there any report about modulated structure happend in ZrO2?
We have a Specimen Injection Rod and 4, PW 6145/00 3 mm. sample holders for an Philips EM 300 Biological Microscope available. Are far as I know, these have never been used because there is no evidence of contamination or wear on the rod or holders.
If you are interested, make an offer.
Fred Pearson McMaster University Brockhouse Institute for Materials Research Hamilton Ontario Canada L8S 4M1
We have a Specimen Injection Rod and 4, PW 6145/00 3 mm. sample holders tips for a Philips EM300 Biological Microscope for sale.
Are far as I know these have never been used because there is no evidence of contamination or wear on the rod or holder.
If you are interested, make an offer.
email: eoptics-at-mcmail.CIS.McMaster.CA
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
I own an old Leitz Wetzlar bi-microscope, and have no manual for maintenance, instructions, etc. Anyone here able to provide some pointers or reference? Many thanks. Frank G Anderson 745 Sipsiri Soi 3 Myang, Korat 30000 Thailand
Miguel Angel S=E1nchez wrote: } =20 } -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } -----------------------------------------------------------------------. } =20 } Hi there: } My name is Miguel Sanchez } my problem is the following } As part of my business I receive } micron rated textiles, that is nylon mesh } that has a determined distance between } threads. } Sometimes is very difficult to determine } that the textile is inside tolerances or } not. } I use an optical microscope to do measure them } my question is: } What do I need to measure the textiles } with the use of a camera and a computer? } I want to use a camera so we can agree } on what we are measuring. } The solution has to be cost effective. Very cost } effective. } Your comments will be highly appreciated } Thanks You need a reticule and a stage micrometer. The reticule inserts into an eyepiece and provides a grid. The stage micrometer allows for measurement of each increment.=20
You can obtain from Klarmenn Ruling, Inc. Manchester NH (603) 424-2401.
You'll need to know the ID of the eyepiece and what measurement scale you want to use.
Seth; Ted Pella offers Printerface for Windows, an image capture system that generates digital images from analog SEM images (I think the board is manufactured by Gatan). I haven't tried it yet, but I've heard it works well. (I have no financial interest in Pella or Gatan). Pella's phone # is (800) 637-3526 (USA).
Leslie Eibest Zoology Dept., Box 90325 Duke University (919) 684-2547 leibest-at-duke.edu
Sanford Simon wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anyone know of a source for teflon o-rings with a thickness of {100 uM? } } Thanks, } Sandy Simon } } Sanford M. Simon } Laboratory of Cellular Biophysics } Box 304 } Rockefeller University } 1230 York Avenue } New York, N.Y. 10021 } 212-327-8130 (voice) } 212-327-8022 (fax) } simon-at-rockvax.rockefeller.edu (e-mail) You can try Roger Zatkoff Company -at- (810) 478-2400, Farmington Hills, MI. They carry nothing but O Rings.
Bob Baier at the Univ of Buffalo apparently saw my name on this list, and sent me a personal message about a week ago. I have since tried several times to reply to him using the address: baier-at-ubvms.cc.buffalo.edu - (which I copied directly from the original message I received from him), but keep getting rejections due to 'local delivery' problems. Does anyone happen to know a better address for him, or have suggestions on how to get through to him with this one?
TIA
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Chapter 3 of the book "Advanced Scanning Electron Microscopy & X-ray Micro Analysis" by Newberry, Joy, et. al, Plenum Press 1986 is entitled "Electron Channeling Contrast in the SEM", and ought to provide the information you are looking for.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I'd be pleased to hear from people who are working with the BAL-TEC BAF-060 freeze-etch system, if nothing else, to swap stories, experiences and ideas. I'm thinking about modifying or redesigning the double replica specimen table, because the BAL-TEC (Technotrade) model looks too difficult to load without pre-fracturing the sample. Has anyone had good luck with the off-the-shelf model? or come up with a better design? The holders with 3 individual sample slots used on older Balzers systems seemed to work well, but to my knowledge that design is not available for the BAF 060.
Thanks for any feedback, Tom
Thomas H. Giddings Tel: (303) 492-8402 MCDB Electron Microscopy Service Fax: (303) 492-7744 University of Colorado giddings-at-spot.colorado.edu Boulder, CO 80309-0347
Dear Glenn, From what I can remember, the few times I have see channeling contrast on the SEM, it is what shows up when other sources of contrast are missing. The sample must be electropolished (to eliminate surface deformation), smooth (to eliminate SE contrast), homogeneous (to eliminate BS contrast), then crank the contrast way up. I think the BS detector works best. High voltage helps. Albert Curzon of the Physics Department of Simon Fraser University in Burnaby, B.C. Canada has done SEM studies of magnetic domain, which is imaged in a similar way. You wrote:
} } Can anyone refer me to a basic reference on electron channeling contrast, or failing that, give me a few hints on optimal conditions for observing this type of contrast ? } } TIA } } Glenn } } Glenn Poirier Phone (514) 398 6774 } Electron Microprobe Laboratory Fax (514) 398 4680 } Earth and planetary Science } Mcgill University Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I picked up the following message which was posted to another newsgroup (sci.techniques.microscopy). Anyone who can help, please contact J.L. Beauchamp directly at Caltech.
If you haven't noticed it the Microscopy Listserver was off-line for most of the day today.
This note is just to head off a host of messages asking why people were not able to send/receive Email and/or login to various MSA WWW sites.
Network Operations Center and Ameritek Communications had a major fault in the lines to this site. We finally came back up on-line ~ 11 PM CST . A technical genius at the main switching center basically didn't know what he was doing and incorrectly reconfigured a central router about 11 AM this morning. It took lots of phone calls to get it sorted out.
All the local systems here were running, they just could not talk to anyone on the outside world. Well not quite true, I still managed to reconfigure my backup modem connections and send a couple of messages...
AccuMed International, Inc., has an immediate opening for a technical = manager to oversee scientific and technical projects involved in the = development, scientific and clinical testing, and production of = computerized medical laboratory instruments and software-based = applications. The successful candidate will carry out day-to-day = activities involved in shepherding new instruments from R&D to finished = product. The position requires a minimum of a Bachelor's degree and a = strong scientific/technical background, with at least 3-5 years = experience in developing, marketing or supporting imaging workstations = and software-based applications for light microscopy. Specific areas of responsibility will include: - managing the development of new hardware and software products - establishing and maintaining timelines for delivery of prototype = instruments and finished products, including software-based applications - coordinating activities in various departments including R&D, = engineering, software development and manufacturing - assisting with applications to regulatory agencies
AccuMed International, headquartered in Chicago, is a global laboratory = diagnostic products company and an emerging leader in the field of = automated cytopathology. EOE.
Please respond via mail, FAX or email (NOT telephone) to:
Marc M. Friedman, Ph.D. Director, Scientifc and Technical Affairs AccuMed International, Inc. 900 N. Franklin, Suite 401 Chicago, IL 60610 Tel: (312) 642-9200 FAX: (312) 642-8684 email: marc.friedman-at-accumed.com
It is not uncommon for me to employ electron channelling contrast. Almost all of my experience is with stainless steels, nickel superalloys (Inconels), Zirconium alloys, and uranium compounds. The ecc is a weak signal and is not always easy to produce.
A few ideas....
Use BSE imaging with large beam currents and high BSE signal gain (contrast). It may be necessary let any response from areas of higher or lower Z than the matrix of interest go saturated black and/or white to achieve this.
Use a "normal" incident beam (0 degrees tilt)
Contrary to some advice I have recieved, I find that lower beam voltages (10 kv) work better than higher (20-30 kV). I have no proof, but suspect that the lower penetration depth images the surface grains without "confusing and diluting" the image with BSE returns from sub-surface grains with different orientations.
Surface preparation is VERY important. A very well polished surface, free from surface damage is required or the signal will be obscured. Some materials are easier than others to prepare. On occation, I have had to send samples back to our met-lab several times before a satisfactory surface is available. A very light "attack" polish (not really an etch) can be helpful during final polish for some materials. I have wanted to try a light ion beam cleaning, but don't have one.
Incident beam angle (with specimen at 90 nominal) changes resulting from the
raster can strongly affect contrast. Experiment with working distance vs. magnification to see what works best for you. Along this line... Don't expect to be able to generate multiple image mosaics which match well. A grain that is dark on one edge of an image may will be light when the stage is translated one frame over because the incident angle is nolonger the same.
Hope this helps...
Woody White
http://www.geocities.com/capecanaveral/3722
BTW something happened to my email pgm half way through??? Hope this dosen't "run over" line length too much!
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Good afternoon everyone.
Can anyone refer me to a basic reference on electron channeling contrast, or failing that, give me a few hints on optimal conditions for observing this type of contrast ?
TIA
Glenn
Glenn Poirier Phone (514) 398 6774 Electron Microprobe Laboratory Fax (514) 398 4680 Earth and planetary Science Mcgill University
THERE ARE THREE SIDES TO EVERY STORY: yOURS, MINE AND THE TRUTH
} Date: Tue, 25 Mar 1997 11:46:20 -0500 (EST) } From: Michael Knotts {ph281mk-at-prism.gatech.edu} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Looking for veteran SEM service engineers } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello Microscopists, } } Can anyone suggest names of service engineers with experience on } vintage Cambridge electron microscopes? I have a 1971 Cambridge } Stereoscan S4 SEM in need of service. I'd like to locate an } experienced technician in the metro Atlanta area or the southeastern } US, but I would appreciate all references and information. Thanks, } } Michael Knotts } ------------------------------------------------------------- } Michael E. Knotts, Ph.D. E-mail: ph281mk-at-prism.gatech.edu } Contributing Editor {The Light Touch} OPTICS & PHOTONICS NEWS } Georgia Tech / School of Physics / Atlanta, GA 30332-0430 } Tel: (404) 894-3422 FAX: (404) 894-9958 }
Yes, Hugh Whitaker (WHIT) used to keep our ancient Cambridge Stereoscan going with corks and paper clips from the local electronics store when he couldn't get parts from Cambridge. I think he is retired now, but he is a wealth of information, and being retired, may be available to help you out. He used to live in Raleigh, but I think he may have moved to the coast of NC. His daughter, Sharon Drew, works at Medical Univ. Hosp. in Charleston 803 792 4157. Perhaps she can get in touch with him for you. Sara
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} Talking of peak overlaps, is anyone out there capable of reliable } accurate analysis of V in the presence of large amounts of Ti, using: } } a WDS } } b EDS? } } I ask because I concluded recently that I couldn't, with my EDS-only } system, analyse for V in titanomagnetites, and that it probably } wasn't possible even with WDS. } Ritchie } } Ritchie Sims phone: 64 9 3737599 ext 7713 } Department of Geology fax: 64 9 3737435 } University of Auckland } Private Bag 92019 } Auckland } New Zealand
EELS on a TEM, with energy resolution around 1-2 eV, pretty easily separates Ti and V, using the L edges, and there is little problem of confusion with N K edge. Accuracy of quantitation won't be very good but it might allow you to decide if you really do have a small amount of V in Ti.
However, what is known as 'Sod's Law' ensures that the O-K edge sits almost exactly at the energy of the V-L edge! If you have strongish V (or O) edges, then you can distinguish O-K from V-L on the basis of edge shape, but if you've got that level of V, you can probably separate V and Ti by EDX anyway!!
Seth, for your optical microscope image capture, we have a program called "Image Central" that will archive and database all of your digital images and let your acquire them directly from a variety of sources. Please contact me fro further info and I will send a package of literature out to you.
Scott E. Berman Advanced Imaging Concepts, Inc. Princeton, NJ Phone: (609) 921-3629 x26 Fax: (609) 924-3010 e-mail : Scott E57-at-aol.com
I have just read an artical that includes in its protocol for fluorescent dyeing, blot drying of the samples, then drying under a stream of nitrogen.
My question was what is the benefit of drying under a stream of nitrogen as opposed to air drying? My first thought was perhaps to reduce any oxidation. Can anyone shed any light on this matter.
Thanks,
Chris. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Chris Johns Queensland University of Technology 2 George St. Brisbane QLD 4001. Email: {c.johns-at-student.qut.edu.au} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Please find hereafter the major characteristics of our grabbing system for SEMs: Orion 4.2 for Windows.
=B7 Digitizes in any size between 16x16 and 4096x4096 pixels (depending on your SEM) =B7 Square pixels in any mode =B7 Connects to any existing SEM or STEM =B7 100% reliable for your SEM and lowest noise level due to unique, optical isolation on board =B7 Grabs images in slow scan and full photo resolution =B7 Can grab 2 simultaneous images (for example SE and BSE fields) =B7 Unique, programmable oversampling factor dramatically reduces image= noise in real time =B7 The grabbed images are immediately available to the user in the PC RAM =B7 They are stored in full resolution, 256 grey levels per pixel (frame integration uses 16 bits per pixel during grabbing) =B7 Easy transfer to any other application (OLE 2 or clipboard) =B7 Distance measurement, extraction, filtering, line / rectangle drawings, line scan function =B7 Selectable, enhanced image compression engine reduces image size on disk by 90% with little image degradation =B7 Full compatible with Windows 3.1x, Windows 95 and Windows NT.
Options =B7 EDX mapping grabbing mode mixes elemental maps with SE or BSE image =B7 Ultra high resolution photo replay allows image to be photographed later from disk =B7 HP Laserjet enhancement card allows photographic quality in 10 seconds =B7 Powerful yet simple programming language makes the user=92s job easier
See also our WEB site http://www.microscopy-uk.org.uk
At 16:33 24/03/1997 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Best regards,
Paul Vanderlinden. Sales Manager.
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D See our web site: http://www.microscopy-uk.org.uk =20
} } Does anyone know of a source for teflon o-rings with a thickness of {100 uM? } } Thanks, } Sandy Simon } } Sanford M. Simon } Laboratory of Cellular Biophysics } Box 304 } Rockefeller University } 1230 York Avenue } New York, N.Y. 10021 } 212-327-8130 (voice) } 212-327-8022 (fax) } simon-at-rockvax.rockefeller.edu (e-mail)
We purchased teflon o-ring 9 years ago from Bal Seal Engineerig Co., 620 W. Warner Ave. Santa Ana, CA 92707-3398. Tel: 714-557-5192.
Ya Chen
Ya Chen
========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
I request your help in locating a publisher or bookseller who carries the book "Practical electron microscopy in Material Science" by J.W.Edington. It was originally published as a 4 vol. series by Philips. Van Nostrand who published the combined volume in 1976 say the book is out of print.
Thanks for your help.
Mohan Kalyanaraman Sr. Staff Material Scientist Mobil Technology Company Paulsboro, NJ 08066 609-224-3989
Chapter 3 of the book "Advanced Scanning Electron Microscopy & X-ray Micro Analysis" by Newberry, Joy, et. al, Plenum Press 1986 is entitled "Electron Channeling Contrast in the SEM", and ought to provide the information you are looking for.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Bob Baier at the Univ of Buffalo apparently saw my name on this list, and sent me a personal message about a week ago. I have since tried several times to reply to him using the address: baier-at-ubvms.cc.buffalo.edu - (which I copied directly from the original message I received from him), but keep getting rejections due to 'local delivery' problems. Does anyone happen to know a better address for him, or have suggestions on how to get through to him with this one?
TIA
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
There are a few vendor that offer Image capturing for the SEM. Most of the system we have installed on our refurbished SEM are supplied by Evex Analytical. There number is 609-252-9192.
I am trying to find out what is the resolution of a Confocal Scanning Laser Microscope. Does it depend on the type of laser used?
I also have another question concerning SEM microscopes. Why is the column of the SEM shorter than the TEM. It is because of a simpler electron optic system in the SEM?.
I know that these are basic questions, but answers to questions like these are impossible to find in textbooks
} Dear All, } } I request your help in locating a publisher or bookseller who carries } the book "Practical electron microscopy in Material Science" by } J.W.Edington. It was originally published as a 4 vol. series by } Philips. Van Nostrand who published the combined volume in 1976 say } the book is out of print. } } Thanks for your help. } } Mohan Kalyanaraman } Sr. Staff Material Scientist } Mobil Technology Company } Paulsboro, NJ 08066 } 609-224-3989
Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility ------------------------------------------------------------------- Pereant qui ante nos nostra dexerunt.
Hope I am not double posting... The orginal didn't show up and may have been lost is cyberspace...??? _W_
It is not uncommon for me to employ electron channelling contrast. Almost all of my experience is with stainless steels, nickel superalloys (Inconels), Zirconium alloys, and uranium compounds. The ecc is a weak signal and is not always easy to produce.
A few ideas....
Use BSE imaging with large beam currents and high BSE signal gain (contrast). It may be necessary let any response from areas of higher or lower Z than the matrix of interest go saturated black and/or white to achieve this.
Use a "normal" incident beam (0 degrees tilt)
Contrary to some advice I have recieved, I find that lower beam voltages (10 kv) work better than higher (20-30 kV). I have no proof, but suspect that the lower penetration depth images the surface grains without "confusing and diluting" the image with BSE returns from sub-surface grains with different orientations.
Surface preparation is VERY important. A very well polished surface, free from surface damage is required or the signal will be obscured. Some materials are easier than others to prepare. On occation, I have had to send samples back to our met-lab several times before a satisfactory surface is available. A very light "attack" polish (not really an etch) can be helpful during final polish for some materials. I have wanted to try a light ion beam cleaning, but don't have one.
Incident beam angle (with specimen at 90 nominal) changes resulting from the
raster can strongly affect contrast. Experiment with working distance vs. magnification to see what works best for you. Along this line... Don't expect to be able to generate multiple image mosaics which match well. A grain that is dark on one edge of an image may will be light when the stage is translated one frame over because the incident angle is nolonger the same.
Hope this helps...
Woody White
http://www.geocities.com/capecanaveral/3722
BTW something happened to my email pgm half way through??? Hope this dosen't "run over" line length too much!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good afternoon everyone.
Can anyone refer me to a basic reference on electron channeling contrast, or failing that, give me a few hints on optimal conditions for observing this type of contrast ?
TIA
Glenn
Glenn Poirier Phone (514) 398 6774 Electron Microprobe Laboratory Fax (514) 398 4680 Earth and planetary Science Mcgill University
THERE ARE THREE SIDES TO EVERY STORY: yOURS, MINE AND THE TRUTH
} } I also have another question concerning SEM microscopes. Why is the } column of the SEM shorter than the TEM. It is because of a simpler } electron optic system in the SEM?. } } I know that these are basic questions, but answers to questions like } these are impossible to find in textbooks } } Thanks. } } GCH.
Hi Gary,
Basicly, a TEM has condenser, objective, intermediate, and projection lenses, plus a fluorescent screen and a film chamber. In contrast, SEM has only condenser and objective lenses, plus a speciemen chamber. That is obviously why SEM has a short column.
Ya Chen
Ya Chen
========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
The Integrated Microscopy Resource and Carnegie Mellon University will be sponsoring a symposium and short course on multi-photon excitation imaging, August 9-10, 1997, in Cleveland Ohio.
} I ask because I concluded recently that I couldn't, with my EDS-only } system, analyse for V in titanomagnetites, and that it probably } wasn't possible even with WDS. } If I ask my system to look for V in my standard Rutile (TiO2), it } always finds a few tenths of a %, even though my software } (LINK ZAF4/FLS) strips out Ti Kb along with the Ka. I have no way of } telling whether this V is genuine or not, but I suspect that it } isn't, as my supposedly-pure Ti metal standard also gets credited } with a small amount of V, which I doubt. } I was recently given another rutile standard, and, blow me down, it } is reputed to have 0.4% V by WDS analysis. } Anyone got any feeling for whether this is likely to be true?
The general case for interference with the transition elements is that the Z-1 element Kb peak overlaps the Z element Ka peak. Ti Kb on V Ka is what you bring up.
For EDS analysis, assuming that linear least squares deconvolution is used, the fit for Ti should handle both Ti Ka and Kb if they are grouped together as your EDS reference. Any residual could then appear as a positive value for V. This may reflect a calibration difference between your sample and reference spectra (I'm assuming we are not talking standardless EDS here).
For WDS measurement one has the choice of PET or LIF; the latter has better resolution and should be used to reduce the magnitude of the overlap to begin with. I find that (using LIF) the measured V Ka k-ratio on synthetic (pure) TiO2 is typically less than 0.5%, which compares with:
} I was recently given another rutile standard, and, blow me down, it } is reputed to have 0.4% V by WDS analysis.
My guess is that the analyst did not correct for the overlap. In the absence of any more sophisticated software, you can make a correction for a simple overlap like this via:
corrected V K-ratio = measured V K-ratio - AK * measured Ti K-ratio
where AK is the apparent V K-ratio measured on pure TiO2. Thus, when analyzing TiO2 the resulting V concentration is zero, and when analyzing a phase containing no Ti the correction is zero. This method corrects the K-ratio before passing to the ZAF program, but you could just as well make a small correction like this on the weight percent data. In practice I find that this linear correction reduces the apparent V in pure TiO2 by an order of magnitude, i.e. from 0.X% to 0.0X%, so especially several hundred ppm V in rutile should be viewed with scepticism.
You can also do a wavelength scan on your rutile looking for the presence of the V *Kb* peak, since this is not overlapped. If you see this peak, then you know there is V in the sample.
This is not a bad overlap. Consider Pb La on As Ka using LIF. This is a total overlap with no real possible solution via this linear correction. The solution in a case like this is to use As Kb and either Pb Lb or Pb Ma as your lines (the application is for sulfides or arsenides). As long as there are no absorption edges between the Ka and Kb lines, or La and Lb lines, the mass absorption coeffiecients are usable; one really has no other choice.
Have fun,
Paul
+----------------------------------------------------+ | Paul K. Carpenter paulc-at-gps.caltech.edu | | Division Analytical Facility | | Geological and Planetary Sciences MC 100-23 | | California Institute of Technology | | Pasadena, CA 91125 | | 818-395-6126 (X-ray Lab) 818-568-0935 (Dept. FAX) | +----------------------------------------------------+
} I have just read an artical that includes in its protocol for fluorescent } dyeing, blot drying of the samples, then drying under a stream of nitrogen. } } My question was what is the benefit of drying under a stream of nitrogen as } opposed to air drying? My first thought was perhaps to reduce any oxidation.
Oxidation is one consideration. The lack of water vapor in the N2 stream is another, and lack of dust, oil droplets, etc. in N2--these are often present in compressed air--is a third.
} Can anyone shed any light on this matter. } If I did, would you just send it back longer? ;-) Yours, Bill Tivol
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My LEO service engineer recommended that I contact you to get an e-mail address for Chris Martinic at the University of New South Wales. Chris has information about image grabbers that I need. Please send me his e-mail address. My address is:
tom.cox-at-lmco.com
Thank you,
Thomas J. Cox Lockheed Martin Missiles & Space Co.
I would like to know what goes on inside the chamber of my electron microprobe when the focused, moderate to high current (10 - 200 nA) electron beam hits epoxy. Visually, a crater forms. What are the solid/gaseous by-products of that reaction? Any ideas, educated guesses, and/or experimental or theoretical results (or suggestions where to look) would be appreciated. Thanks.
John
John Fournelle Electron Microprobe Lab office: (608) 262-7964 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA/9 Personal http://geology.wisc.edu/~johnf/ Probe lab http://geology.wisc.edu/~johnf/sx51.html
"The first rule of all intelligent tinkering is to save every cog and wheel." Aldo Leopold
} I am trying to find out what is the resolution of a Confocal Scanning } Laser Microscope. Does it depend on the type of laser used? } Dear Gary, The resolution of a CSLM is ~1/3 to ~1/2 the wavelength of the ilumination in the x-y plane and about 2x worse in the z-direction. Jim Pawley's Handbook of Confocal Microscopy has a very good description of the theory. (I hope I got the title right--the book is at my desk & I'm not.) It will depend on the wavelength of light used, but not on whether the laser is pulsed or continuous wave, so whether it depends on the "type of laser used" depends on what you mean. There may be CSLM's which use frequency-doubled light, so a pulsed laser is neces- sary to provide sufficient intensity to get the non-linear optics to work, so if this is what you mean, then whether the CSLM works at all can depend on the type of laser used, but once you get light of a par- ticular wavelength, the resolution won't depend on where that light came from. Since I'm not an expert on this, you should probably check the book. Yours, Bill Tivol
} } Hello Microscopists, } } } } Can anyone suggest names of service engineers with experience on } } vintage Cambridge electron microscopes? I have a 1971 Cambridge } } Stereoscan S4 SEM in need of service. I'd like to locate an } } experienced technician in the metro Atlanta area or the southeastern } } US, but I would appreciate all references and information. Thanks,
Clark Houghton at Secondary Images is a real wizard with the old Cambridges. He is in Ohio (but was willing to come all the way to Hawaii... what a hardship!) at (513) 927-5373.
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Hello! We have a Cambridge Stereoscan and for service use Scanners Corporation in Eldersburg, MD - tel. 1-410-549-3800 I believe they cover the East Coast and possibly a bit further out as well. They take great care of our SEM. Nancy
In response to the question as to why SEM colums are so much smaller than those of TEMs, I offer the following:
The colums of SEMs basically contain only two lenses: a 'first' lens (often incorrectly called the condenser lens) which produces demagnification of the spot from the electron gun, and a 'second' lens (usually incorrectly called the objective lens) which is used to focus the spot image produced by the first lens onto the specimen forming the electron probe which generates the signals that are emitted from the sample. Sometimes the first lens is a compound lens, actually giving a total of three lenses; nonetheless, they function as a two-lens system overall.
This entire electron optical system of an SEM corresponds in general character and function to the 'condenser lens system' (i.e. the system of lenses that is above the specimen chamber) of a TEM. That is, the condenser lens system of a TEM is basically a two-lens system which functions to control the illumination that strikes the specomen (just as the two lens system of the SEM does).
In a TEM; however,there is an additional image-forming lens system below the specimen consisting of the objective lens, which produces the primary electron image from the specimen, and then a series of from 3 to 6 additional lenses which produce the wide range of magnifications we expect from TEMs now-a-days, plus providing the variety of additional imaging functions commonly available (selected area diffraction, convergent beam diffraction, etc.). No lens is directly involved in the image-forming process in an SEM, and so these additional lenses are not needed.
Thus there are three or four times as many lenses in a TEM as there are in an SEM. In addition, whereas most TEMs are designed to operate at electron accelerating voltages from 80 kV to several hundred kilovolts, most SEMs do not use accelerating voltages much above 30 kV. Since it is much easier to deflect these lower energy electrons, the magnetic fields in SEM lenses do not need to be as strong as those in TEMs; consequently they require fewer turns of wire in their excitation coils, and so can be physically much smaller.
I hope this answers your question satisfactorily. If not, don't hesitate to let me know. I can send you column diagrams which illustrate the above points, if needed.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
In response to the question as to why SEM colums are so much smaller than those of TEMs, I offer the following:
The colums of SEMs basically contain only two lenses: a 'first' lens (often incorrectly called the condenser lens) which produces demagnification of the spot from the electron gun, and a 'second' lens (usually incorrectly called the objective lens) which is used to focus the spot image produced by the first lens onto the specimen forming the electron probe which generates the signals that are emitted from the sample. Sometimes the first lens is a compound lens, actually giving a total of three lenses; nonetheless, they function as a two-lens system overall.
This entire electron optical system of an SEM corresponds in general character and function to the 'condenser lens system' (i.e. the system of lenses that is above the specimen chamber) of a TEM. That is, the condenser lens system of a TEM is basically a two-lens system which functions to control the illumination that strikes the specomen (just as the two lens system of the SEM does).
In a TEM; however,there is an additional image-forming lens system below the specimen consisting of the objective lens, which produces the primary electron image from the specimen, and then a series of from 3 to 6 additional lenses which produce the wide range of magnifications we expect from TEMs now-a-days, plus providing the variety of additional imaging functions commonly available (selected area diffraction, convergent beam diffraction, etc.). No lens is directly involved in the image-forming process in an SEM, and so these additional lenses are not needed.
Thus there are three or four times as many lenses in a TEM as there are in an SEM. In addition, whereas most TEMs are designed to operate at electron accelerating voltages from 80 kV to several hundred kilovolts, most SEMs do not use accelerating voltages much above 30 kV. Since it is much easier to deflect these lower energy electrons, the magnetic fields in SEM lenses do not need to be as strong as those in TEMs; consequently they require fewer turns of wire in their excitation coils, and so can be physically much smaller.
I hope this answers your question satisfactorily. If not, don't hesitate to let me know. I can send you column diagrams which illustrate the above points, if needed.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
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Not my field, but wavelength of illumination will be one determining factor.
} I also have another question concerning SEM microscopes. Why is the } column of the SEM shorter than the TEM. It is because of a simpler } electron optic system in the SEM?.
Two answers:
1. Voltage - generally, SEMs work at a maximum of 30kV while TEMs typically work from a minimum of 100kV. Higher voltage electron have much greater kinetic energy and are thus more diffcult to deflect. Even with somewhat stronger lenses, the additional energy of the electron requires a little more distance to have the required effect.
2. More significantly, an SEM column is only equivalent to the part of the TEM colum ABOVE the specimen. In a TEM, additional lenses are needed after the specimen to focus the beam into an image. In a SEM, the electron beam is aleady focused to a probe at the specimen, and only needs 'intensity' detectors. Indeed, if you do secondary electron imaging on a bulk specimen in a TEM/STEM type if instrument, the bottom half of the column is not used.
} I know that these are basic questions, but answers to questions like } these are impossible to find in textbooks } } Thanks. } } GCH.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
Seth: we built an SEM image-capture system based on a National Instruments (AT-MIO-16E-10) data-acquisition card, controlled by a program written in Visual Basic. Hardware cost was about $US 3000 including Pentium computer and 1200 dpi laser printer. Brief details of the system will be presented at the June MSC conference, 2-page abstract appearing soon on the conference web page: http://www.ualberta.ca/~mmid/msc
Ray Egerton, Physics Dept, University of Alberta, Edmonton, Canada T6G 2J1 Phone: 403-492-5095, FAX: 403-492-0714, e-mail: egerton-at-phys.ualberta.ca ------------------------------------------------------------------------
I have not had much experience with scanners, but it seem to me the answer to why there columns are not as long as the TEM's are maybe in the fact that you do not have the objective len and stage or the projector or diffraction lens to deal with. Thats my best guess. As for Confocal Scanning I have had no experience at all with this device. sorry!
Dear John, When I did a careful study of SEM sample contamination, using a polished copper sample and carefully regulated kV, current and magnification, the deciding factor seemed to be how recently someone had hit epoxy with the beam. One "hit" seemed to result in much worse contamination for about three days. In high current situations you can see the epoxy "boil". I think the vapourised hydrocarbons fly about your chamber for quite a while. Cleaning with a N2 purge, like the SEMclean system (I use a home-built one), does seem to help. I seem to recall that someone did a study of contamination, air-jets and such in Microbeam Analysis in the early '80s. May have been Bastin. You wrote:
} I would like to know what goes on inside the chamber of my electron microprobe } when the focused, moderate to high current (10 - 200 nA) electron beam hits } epoxy. Visually, a crater forms. What are the solid/gaseous by-products of } that reaction? Any ideas, educated guesses, and/or experimental or } theoretical results (or suggestions where to look) would be appreciated. } Thanks. } } John Hope this helps, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
You might want to check out 4Pi Analysis. they have a a Mac-based image capture system for SEMs that is priced very competitively. There boards and software offer total control over the SEM, with image acquistion by NIH Image, Photoshop, IPLab Spectrum, etc.
Regards, Glen MacDonald Virginia Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
/*---------------------------------------------------------------------- -*/ / The box said "Requires Windows 95 or better.", so I bought a Macintosh. / /*---------------------------------------------------------------------- -*/
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Subject: Time:9:50 AM OFFICE MEMO In Memoriam - Mark Fendorf Date:3/28/97
In Memoriam - Mark Fendorf
It is with great sadness that we at NCEM announce the death of Mark Fendorf.
Mark John Fendorf died on March 14, 1997 at the age of 36. As a collaborative post-doctoral scientist at LBNL's National Center for Electron Microscopy, Mark played a central role in NCEM's outreach to external facility users. He was an outstanding microscopist, a resourceful materials scientist and a delightful colleague to all members of NCEM. In his role of resident expert microscopist he collaborated with many NCEM users and helped countless visitors of the facility. He earned his Ph.D. in materials science from UC Berkeley in 1992. His scientific work focused on electron beam micro-characterization of a number of different materials, including high-temperature superconductors, catalysts, fullerines and most recently the adsorption of heavy metals on soil minerals. He was an active member of several scientific societies, including the Microscopy Society of America and the American Ceramic Society. In 1991, he co-founded the UC Berkeley chapter of the Materials Research Society. Outside of LBNL, he was a dedicated and enthusiastic volunteer for the Boy Scouts of America where he was a leader for 16 years, served on the Monterey Bay Area Council, and acted as Director of Troop Leadership Training for a number of years.
During his progressively debilitating illness, Mark continued his work at NCEM with remarkable dedication and tenacity. His strength of will and his determination to overcome his handicap caused by the illness were admired by all those who knew him. Mark's untimely death has left the Center permanently diminished. He will be greatly missed by his NCEM colleagues and the many users and visiting scientists at the facility.
Mark Fendorf is survived by his parents, Ken and Virginia Fendorf and his brothers Scott and Dale. In recognition of his sustaining enthusiasm for the science of electron microscopy, and in an effort to help others continue where Mark had to leave off, his family has established a memorial fund in support of NCEM's outreach program.
Donations to the Mark Fendorf memorial fund can be made to: World Savings, Aptos Branch, Mark Fendorf Memorial Fund, 7970 Soquel Drive, Aptos, CA 95003
} I would like to know what goes on inside the chamber of my electron microprobe } when the focused, moderate to high current (10 - 200 nA) electron beam hits } epoxy.
When an electron passes through matter, the energy of the electron is transferred to the material in the form of ionizations and excitations. About 30 eV is transferred for each ionization, and most of the energy transferred goes into primary or secondary ionizations. The products, ob- viously far from thermodynamic equilibrium, react with other components of the material causing chemical bond breaking and free radical formation. Finally (a few microseconds later) these reactive species form more stable products, which are often small organic molecules in the case of electrons incident on epoxy resin. There is a statistical distribution of products which depends on the nature of the resin; since many of these are volatile, they will travel throughout the chamber (they also gain some kinetic energy from the energy transferred from the initial electron).
} Visually, a crater forms.
This should make sense in light of the above.
} What are the solid/gaseous by-products of } that reaction? Any ideas, educated guesses,
The above are educated guesses...
} and/or experimental or } theoretical results
and theoretical results.
} (or suggestions where to look) would be appreciated.
My info comes from Friedlander, et al., Nuclear and Radiochemistry; I do *not* recommend this book. A good book on radiation chemistry would be a good place to start, but I don't know any titles. Yours, Bill Tivol
Jeol sells a high resolution standard (gold on carbon) that we would like to purchase. Does anyone have info: (part number, price, where to order)? Many thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 formal proposal to create a new group tentatively called sci.bio.immunocytochem was posted to news.announce.newgroups on 17.3.97. This is a reposting of that proposal. Please have a read and let me know what you think.
REQUEST FOR DISCUSSION (RFD) unmoderated group sci.bio.immunocytochem
This is the 2nd Request For Discussion (RFD) for the creation of a world-wide unmoderated Usenet newsgroup sci.bio.immunocytochem, currently being discussed in news.groups
Suggestions for improvements to this proposal are welcome. Discussion about it should take place in news.groups. A vote is expected to be held in about three to four weeks.
This is not a Call for Votes (CFV); you cannot vote at this time. Procedural details are below.
CHANGES from previous RFD:
2nd RFD posted because more than 60 days have elapsed since 1st RFD. There are (minor changes in Distribution and Newsgroup line.
Newsgroup line: sci.bio.immunocytochem Immuno-labelling of biological material.
RATIONALE: sci.bio.immunocytochem
Immunohistochemists and immunocytochemists already enjoy the benefits of online communication, utilizing e-mail, accessing web sites, and subscribing to specialised mailing lists. Usenet newsgroups are also popular, but this is less obvious because articles with immunocytochemical/immunohistochemical content get posted to many different newsgroups. Most articles are posted to a favourite five or six newsgroups including bionet.cellbiol, sci.med.immunology and sci.techniques.microscopy, but often articles get posted to any one of fourteen or fifteen newsgroups in the sci. and bionet. heirarchies. Some of these are listed in the distribution list at the end of this proposal.
In my view, no existing newsgroup fulfils the criteria necessary to attract all the various immunocytochemistry postings. I do not wish to draw users away from other newsgroups, only to encourage scientists to share their knowledge and expertise on immunocytochemistry in the most effective manner. In response to my proposal to create a newsgroup dedicated to the discussion of immunocytochemistry and immunohistochemistry, I have received e-mail and faxes from researchers all over the world offering their support and encouragment.
Immunocytochemistry and immunohistochemistry are not subdivisions of immunology, molecular biology or chemistry. Microscopy, although essential, is only a small part of the story. Immunocytochemistry and immunohistochemistry are multi- disciplinary, therefore discussions are destined to stay distributed amongst the different newsgroups until they are all brought together under one umbrella. This would then act as a focus point for all the immunocytochemists who are already Internet users, and encourage new subscribers to Usenet.
CHARTER: sci.bio.immunocytochem
This is a newsgroup for the exchange of information relating to immunocytochemistry and immunohistochemistry. This unique research tool is used to locate and identify specific molecules in biological material, at the microscopical level.
Articles posted to this group must be relevant to one or more aspects of the above. The kind of subjects that may be discussed include techniques, theory, presentation of results, requests for collaboration, history, equipment, publication references, notice of events, tips and trouble-shooting, jobs offered andwanted, jokes, stories and new ideas, so long as the posting bears a direct relevance to the central theme. There will be a list of Frequently Asked Questions (FAQs) to help newcomers.
A relevant posting could just be a simple question or answer, for example "Has anyone got any experience with this reagent ?"or "Which course could I attend to learn more about immunogold labelling?". There will be articles reminding people to read the list of FAQs prior to posting their own article. Usenet readers may get involved in complex discussions about, for example, multiple labelling, proper use of control experiments, microwave antigen retrieval or quantitative measurements. Remember that articles posted to a newsgroup are intended for a wide readership, so if you have information which concerns only one or two people then please don't use this newsgroup, use e-mail.
Commercial advertisements for services, equipment or reagents violate the charter unless one or more of the following apply: (a)The advertisement is part of a comprehensive article designed specifically to address issues raised in earlier articles posted to the group (b)A general reference to the type of product does not suffice for technical reasons and it is necessary to specify the exact commercial product (c) The information is offered primarily for the benefit of the readers (d)The advertisement is for second-hand equipment specific to immunocytochemistry (e) Requests or offers for free products are acceptable if they are not part of a sales promotion.
END CHARTER.
PROCEDURE:
This is a request for discussion, not a call for votes. In this phase of the process, any potential problems with the proposed newsgroups should be raised and resolved. The discussion period will continue for a minimum of 21 days (starting from when the 2nd RFD for this proposal is posted to news.announce.newgroups), after which a Call For Votes (CFV) may be posted by a neutral vote taker if the discussion warrants it. Please do not attempt to vote until this happens.
All discussion of this proposal should be posted to news.groups. This RFD attempts to comply fully with the Usenet newsgroup creation guidelines outlined in "How to Create a New Usenet Newsgroup" and "How to Format and Submit a New Group Proposal". Please refer to these documents (available in news.announce.newgroups) if you have any questions about the process.
DISTRIBUTION:
This RFD has been posted to the following newsgroups: news.announce.newgroups,news.groups,bionet.cellbiol, bionet.diagnostics,bionet.immunology,bionet.microbiology, bionet.molbio.methds-reagnts,sci.bio.misc,sci.med.immunology, sci.techniques.microscopy
This RFD will be reposted to the following newsgroups after its posting in news.announce.newgroups: bionet.molbio.proteins,bionet.neuroscience,bionet.plants, sci.bio.microbiology,sci.med,sci.med.laboratory,sci.misc, sci.nanotech
This RFD will also be reposted to the following mailing lists after its posting in news.announce.newgroups:
Histonet mailing list: {histonet-at-pathology.swmed.edu} Information pertaining to the technical aspects of histology and histopathology such as tissue fixatives and processing, routine histology, special stains, immunohistochemistry, in-situ hybridization etc. To subscribe type "subscribe digest" into the subject box and leave the text box empty, or to subscribe to the full service just type "subscribe". For more info access web site http://www.mwrn.com/subject/histonet.htms
Microscopy Society of America listserver: Questions/comments/answers in the various fields of Microscopy Currently over 3000 subscribers. To subscribe send the message "subscribe" to {Listserver-at-MSA.Microscopy.Com} then send messages in plain text to {Microscopy-at-MSA.Microscopy.Com} For more info access web site http://www.amc.anl.gov/ Docs/anl/Nestor/Software/telecommList.html
Stanford University list server To subscribe, send a message to {majordomo-at-pathology.stanford.edu} with "subscribe ipox-l" in the body of your message. This list helps pathologists and other laboratory professionals to exchange information about immunoperoxidase methods.
This RFD will also be reposted to the following web-sites after its posting in news.announce.newgroups:
Royal Microscope Society http://www.rms.org.uk Web Master Dr R. A. D. Mackenzie {r.a.mackenzie-at-open.ac.uk}
Center for Cell Imaging Department of Cell Biology Yale University School of Medicine Introduction to Immunocytochemistry http://info.med.yale.edu/cellimg/CCIimmuno.html Web Master Paul Webster { paul_webster-at-yale.edu}
Proponent: Amanda Wilson {awilson-at-aw.u-net.com} Proponent: Paul Monaghan { monaghan-at-icr.ac.uk} Mentor: Jonathan Grobe {grobe-at-netins.net}
simple question, when recharging a balzers gun with platinum, where should the tip of the carbon rod holding the platinum pellet be with repect to the tungsten coil?
Tx
simon
-- ------------------------------------------------- Simon C. Watkins Ph.D Associate Professor Director Center for Biologic Imaging University of Pittsburgh Pittsburgh PA 15261 tel 412-648-3051 fax 412-648-2004 -----------------------------------------------
I'm about to attempt UV polymerization of Unicryl for the first time and need advice about lamps, embedding molds which are UV penetrable, etc., etc. Virtually no specific advice is given in the circular I received or in the two papers referenced in this literature. I would appreciate any tips, and/or references from any experienced user. Thank you. Grace Kennedy
Luc Nocente wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Try Visilog from Noesis Vision Inc, we have a turn key system available for } sperm analysis and or fibre analysis. You can reach us at } http://www.noesisvision.com } } At 12:51 PM 3/7/97 +0530, SONEJA A K wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } } } Hello guys, } } } } Looking for image analysis software/solutions for the following: } } } } 1} sperm analysis,cell motility etc. } } 2} textile analysis,rayon fibre analysis,broken fibre etc. } } } } Could anyone point out some good manufacturers/sources with } } address/email/fax wwith name of source.?////// } } I would be grateful if someone could help me in this context. } } } } Thanks, } } } } Best regards, } } } } Anish } } } } ************************************************************************* } } For further details please contact: } } Soneja A.K. } } Director } } METZER BIOMEDICAL & ELECTRONICS PVT.LTD. } } 327 Wadala Udyog Bhavan,Wadala,MUMBAI(BOMBAY )400 031.INDIA } } Tel:91 22 4145057/4165650 } } Fax 91 22 4168757 } } } } Email:soneja-at-giasbma.vsnl.net.in } } ************************************************************************* } } } } } } ---------------------------------------------------------------------------- } --------------------- } Luc Nocente Tel: 514 345 1400 } Noesis Vision Inc. Fax: 514 345 1575 } e-mail: ln-at-noesisvision.com } 6800 Cote de Liesse, Suite 200 } St-Laurent, PQ } H4T 2A7,Canada } } Visit our new web site at http://www.noesisvision.com } ---------------------------------------------------------------------------- } ---------------------Dear Anish,
A belated response to your message of 3/7 re: image analysis for sperm motility/analysis:
We did some work recently with Hamilton-Thorne Research. They have several interesting, built-for-purpose systems. Contact: Dr. Dairmid-Douglas Hamilton 100 Cummings Place, Suite 102C 181 Elliott Street Beverly, MA 01915 Phone: (508)921-2050 Fax: (508)921-0250 Please feel free to mention my name.
John- My JEOL parts catalog lists a gold on carbon standard. The part # is 613149. Orders can be sent to JEOL (USA) Inc., Parts Dept., 11 Dearborn Road, Peabody, MA 01960, or call (508) 535-5900 and ask for parts. I don't have a recent price list, so I can't help you there. Leslie
Leslie Eibest Zoology Dept., Box 90325 Duke University (919) 684-2547 leibest-at-duke.edu
We have used a turbo pumped SEM since 1986. Our experience with this system has actually been quite good - we have only had to replace it once. It was fortunate that we are under contract because the pump cost is rather high. The greatest benefits that I see from this system are 1) pump-down time (usually only a few minutes) and 2) cleanliness. One other thing to consider; on our system, the turbo hangs down from a metal bellows connected to the base of the chamber. At one time, we had a very small crack in this bellows, which was not detected for over a year. We replaced numerous ion pumps and other components until it was finally located. Again, fortunately we have always had the system under contract.
-Bob ******************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 PH: (909)399-1311 Email: Bob_Citron-at-cc.chiron.com ********************************
------------------------------------------------------------------------=20 The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20 John Bozzola wrote: =20 Jeol sells a high resolution standard (gold on carbon) that we would like=20 to purchase. Does anyone have info: (part number, price, where to order)?=20 Many thanks. =20 ####################################################################=20 John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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=20 #################################################################### =20 John; You can contact JEOL parts directly in Peabody, MA or if you have the=20 right lab equipment you can make your own. A vacuum evaporator and a=20 30-40 amp. external power supply that can be connected to a second set= =20 of evaporator electrodes is all the equipment that is required. The=20 supplies are standard EM supplies that can ordered from any of the=20 usual sources. If you are interested in making your own I'd be glad=20 to send more details on/off line. =20 John Humenansky Braun Intertec Corp. 6875 Washington Ave. So. Minneapolis, MN 55439 (612) 942-4844 =20
If you have a sputter coater or evaporator, why not make your own. I've had good luck making calibration standards like this (I use them for camera constant calculation, resolution, demos, in TEM). It's good practice in thin film making for students, too ;-)
1. Make a thin carbon film by carbon coating a formvar, butvar etc. coated grid. (how thin the carbon should be depends on the final use; if you want to see the gold lattice in TEM it should be painfully thin). Remove the plastic backing by placing it in a (glass) Petri dish lined with a filter paper pad soaked in CHCl3 (hood).
2. After an overnight stint in the Petri dish, you should have mostly carbon left on the grids.
3. Pop them in your sputter-coater and give them a short blast of Au (we've got a pure Au target) if you use Au/Pd in yours, I'm not sure what you'll get... Well, if you've got a gold target about 1/12 of your normal sputtering time to coat an average SEM sample should work. You'll have a collection of isolated islets of Au. Carbon tape it to a stub and have a look.
Hope this helps, even though it doesn't answer the question.
cheers,
John Heckman TEM supervisor/Center for Electron Optics Michigan State University
Disclaimer: The preceding technique works in my reality; follow my instructions at your own risk!
} -----------------------------------------------------------------------. } } Jeol sells a high resolution standard (gold on carbon) that we would like } to purchase. Does anyone have info: (part number, price, where to order)? } Many thanks. } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901-4402 } 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 } ####################################################################
Hi folks We have a em300 that is about to join the great trashcan in the sky. Before it meets its demise I was wondering whether anyone wants parts from it,
let me know
simon
-- ------------------------------------------------- Simon C. Watkins Ph.D Associate Professor Director Center for Biologic Imaging University of Pittsburgh Pittsburgh PA 15261 tel 412-648-3051 fax 412-648-2004 -----------------------------------------------
I would appriciate hearing from anyone on/off line with experience=20 with turbo pumped SEM's (benefits, horror stories, etc.) =20 Thank you =20 John Humenansky Braun Interec Corp. 6875 Washington Ave. So. Minneapolis, MN 55439 (612) 942-4822
Jeol sells a high resolution standard (gold on carbon) that we would like to purchase. Does anyone have info: (part number, price, where to order)? Many thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
The JEOL resolution standard can be purchased from the JEOL USA parts department. Phone # (508)535-5900. The part number is 613149. I believe this is what you are asking for but verify that with them.
Disclaimer: I have no interest in JEOL
Michael D. Standing e-mail: MDStandi-at-bioag.byu.edu Phone: (801)378-4011
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PHILADELPHIA SOCIETY FOR MICROSCOPY APRIL 1997 MEETING NOTICE
DATE: Tuesday, April 15, 1997
PLACE: Laboratory for Research on the Structure of Matter (LRSM) Building, 33rd and Walnut Street. Parking is available behind the LRSM Building after 5:00 PM.
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7:30 PM Speaker
Chemistry in the Crime Lab Richard Saferstein, Ph.D.
Forensic science in its broadest definition is the application of science to law. As our society has grown more complex, it has become more dependent on rules of law to regulate the activities of its members. Forensic science offers the knowledge and technology of science for the definition and enforcement of such laws. The analytical techniques that can be used in forensic science are numerous and diverse. In general, they must be sensitive enough to cope with minute samples of physical evidence, but they must also be reliable and reproducible to withstand scrutiny by fellow experts in and out of the courtroom. Speed and economy have to be considered too, for the typical forensic scientist must analyze hundreds of cases each year. In this talk a variety of analytical procedures that are applicable to solving forensic science problems will be presented. Included in the discussion will be the application of microscopic analysis to forensic hair and fiber. Also discussed will be the role of visible and infrared microspectrophotometric techniques in forensic problem solving. The speaker will review significant achievements that have been made in utilizing DNA typing for the purposes of linking blood and semen evidence to a single individual. A number of actual case discussions will be included in the talk in order to exemplify the relevancy of forensic science to criminal investigation.
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We have two turbo pumped SEMs - a Leo/Cambridge S-360 (1988) and an Hitachi S-900 (1991). We have never had a problem with either turbo. Local Leo service had a policy of reconditioning the (Pfeiffer) turbos every 3 years for bearing replacement just in case. Their policy now is to leave it until there is a sign of imbalance (audible noise). They are confident now that turbos are no more trouble than any other part so take no special precautionary measures apart from re-oiling the bearing every 6 months. The S-900 has a mag lev. Seiko Turbo. Hitachi advice is that so long as you change the backup batteries every 12 months to ensure full charge (in case of power failure the backup keeps the turbine levitated until it runs down) - the pump will last forever.
Advantage is no fuss clean vacuum. The source of contamination has to be your specimen (or some backstreaming from the rotary, though oils oughtn't get through the turbo if its runnning). AND no problem with water supply/cooling and very little problem with power failure. (Everything coasts to a stop).
BUT a well designed oil diffusion pump system can be NEARLY as good. For that reason I did not insist on the turbo option with our latest Hitachi S-4500. How sure are you of design? Some well known SEMs were plagued with a badly designed diff pump system which stalled and backstreamed with nearly every specimen change!
I am interested in HREM image simulations of III-V semiconductors and I am looking for any references where I can find the values of the Debye-Waller temperature factors for such atoms as P, Ga, IN, As etc. I would be extremely grateful for any helpful information
Sincerely yours
Rafal Spirydon
Dept. of Materials Science and Engineering Kwangju Institute of Science and Technology South Korea
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