Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (esem1-at-traceevidence.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, May 1, 2004 at 08:36:12 ---------------------------------------------------------------------------
Email: esem1-at-traceevidence.org Name: David Spence
Organization: Southwestern Institute of Forensic Sciences
Title-Subject: [Microscopy] [Filtered] MListserver:Looking For Used SEM, EDS and Coating Equipment
Question: I am looking for used SEM, EDS, sputter/carbon coater and related equipment. If you have or know of any equipment that is serviceable and available for haul-off or very inexpensive, please let me know. I am specifically looking for an older quality SEM with a motorized stage that is suitable for automated gunshot residue analysis.
I am involved with a consortium that is being partially funded by the NSF-ATE program. The main project involves the development of 3-D images related to various science, math, engineering and technology areas (from molecules to organs to CAD/architecture) so that they can be projected (stereographic projection). Each institution participating in the project will have a 3-D theatre built so that SMET classes can be brought in for demonstrations of course specific images (the theatre will house a Barco 3-D projector driven by a SGI UNIX-based workstation and a special screen that retains polarization). Students and faculty from the partner institutions will also be trained in a residential program at Brookhaven National Lab by their 3-D Visualization Team.
Since I have been teaching TEM and SEM courses at NCC for almost 20 years, I thought that it would be an interesting project to render student and my own SEM & TEM images for 3-D projection. I have seen many threads on 3-D imaging on this listserver and I would appreciate some advice on where to begin. Initially, I am supposed to identify some software applications that would be suitable for rendering 3-D images of my micrographs. I am aware of some of the applications that are used at BNL for their 3-D theatre such as OpenDX, Open GL and Visualization ToolKit (VTK).
Can anyone suggest an application (UNIX based ideally) that I might get started with? As a novice in this area I realize that the concept of 3-D imaging and stereographic projection are probably two entirely different principles. I would appreciate any references (articles, books, websites) that will help me to understand the concepts.
Thanks for any assistance you can provide!
Steve -- Stephen J. Beck Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
From MicroscopyL-request-at-ns.microscopy.com Sun May 2 15:13:18 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (yuansheng-sun-at-uiowa.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May 2, 2004 at 14:45:18 ---------------------------------------------------------------------------
Email: yuansheng-sun-at-uiowa.edu Name: Yuansheng Sun
Organization: University of Iowa
Education: Graduate College
Location: Iowa city, IA, USA
Question: Dear Madam or Sir:
I am doing a project on deconvolution for 3D phase contrast microscopy. As you know, it is very hard to measure a real psf. So I am wondering how to construct a theoretical psf based on the experimental parameters. Could you give me some infomation on this like some books or papers or technology documents. Thanks! best from yuansheng
I have been using ImageJ and the VolumeJ plugin (for surface rendering) to generate 3D anaglyphs (red/green stereo) as animations. As has been mentioned in this group before, ImageJ is a free program that has been ported from NIH-Image to the Java platform so that it will work on almost any machine. You can get more information on ImageJ at http://rsb.info.nih.gov/ij/
You can download a couple of the anaglyphs at http://astro.temple.edu/~jbs/research/Anaglyphs. Remember that you will need red/green glasses to get the full effect.
Joel
The images were originally obtained as z-series with a Leica confocal microscope.
Date sent: Sun, 02 May 2004 14:17:59 -0400 } From: Steve Beck {becks-at-sunynassau.edu}
I second Joel's suggestion of ImageJ/VolumeJ. You might also want to take a look at VisBio (http://www.loci.wisc.edu/VisBio). VisBio is a open-source JAVA based image analysis tool for multidimensional biological image data. Features include measurement, volume rendering, and arbitrary image stack slicing. Integration with ImageJ is under development.
best regards, kevin
Kevin W. Eliceiri Laboratory for Optical and Computational Instrumentation http://www.loci.wisc.edu Room 271 Animal Sciences 1675 Observatory Drive Madison, WI 53706 Phone: 608-263-6288 Fax: 608-265-3083
From MicroscopyL-request-at-ns.microscopy.com Sun May 2 19:05:12 2004
I guess you know about the GeoWall (http://geowall.geo.lsa.umich.edu/) projects. This is for geologists, but quite amenable to any type of 3D projection really. Somewhat different to the setup you're proposing, because it's PC- or Mac-based, but same principles. There's even a company (VRCO) that sells turnkey systems. There is a bit of background information on the GeoWall site, but more info on some of the link sites.
cheers, Rosemmary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5000 Canberra, ACT 2601 Australia
} From: Steve Beck {becks-at-sunynassau.edu} } Date: Sun, 02 May 2004 14:17:59 -0400 } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] 3-D Imaging & Projection } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Dear Colleagues, } } I am involved with a consortium that is being partially funded by the } NSF-ATE program. The main project involves the development of 3-D } images related to various science, math, engineering and technology } areas (from molecules to organs to CAD/architecture) so that they can } be projected (stereographic projection). Each institution } participating in the project will have a 3-D theatre built so that } SMET classes can be brought in for demonstrations of course specific } images (the theatre will house a Barco 3-D projector driven by a SGI } UNIX-based workstation and a special screen that retains } polarization). Students and faculty from the partner institutions } will also be trained in a residential program at Brookhaven National } Lab by their 3-D Visualization Team. } } Since I have been teaching TEM and SEM courses at NCC for almost 20 } years, I thought that it would be an interesting project to render } student and my own SEM & TEM images for 3-D projection. I have seen } many threads on 3-D imaging on this listserver and I would appreciate } some advice on where to begin. Initially, I am supposed to identify } some software applications that would be suitable for rendering 3-D } images of my micrographs. I am aware of some of the applications that } are used at BNL for their 3-D theatre such as OpenDX, Open GL and } Visualization ToolKit (VTK). } } Can anyone suggest an application (UNIX based ideally) that I might } get started with? As a novice in this area I realize that the concept } of 3-D imaging and stereographic projection are probably two entirely } different principles. I would appreciate any references (articles, } books, websites) that will help me to understand the concepts. } } Thanks for any assistance you can provide! } } Steve } -- } Stephen J. Beck } Professor } Bio-Imaging Center/Electron Microscopy } Department of Biology } Nassau Community College } Garden City, NY 11530 } Voice Mail: (516) 572-7829 } Email: {becks-at-sunynassau.edu} } URL: {http://www.sunynassau.edu/webpages/biology/becks.htm} } }
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 08:17:48 2004
I've just completed a great semester of teaching EM techniques. The student projects that were required are quite interesting. If you have a chance, please review the student's work at this URL:
_________________________________________________ Brian McIntyre Univ. of Rochester The Institute of Optics River Campus EMLab 585-275-3058/4875 585-244-4936 (fax)
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 08:36:45 2004
Please help me. I'm really in trouble. Just burned the second filament after 3 hours. Previously, I had about 200 h. Now the current is very unstable. Before burning, is slowly going down and after that a quick increase up to 170 uA and bang. We have a Jeol 1010 (1 year old).
Many thanks, Luci _________________________________________ Lucian Barbu-Tudoran Electron Microscopy Center Faculty of Biology & Geology Babes-Bolyai University of Cluj-Napoca 5-7 Clinicilor Str., 3400 Cluj-Napoca, Romania Tel/Fax: +40 264 598700 {http://hasdeu.ubbcluj.ro/emc/lucian_barbu_tudoran.htm} http://hasdeu.ubbcluj.ro/emc/lucian_barbu_tudoran.htm
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 08:43:05 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mcintyre-at-optics.rochester.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 08:10:50 ---------------------------------------------------------------------------
Email: mcintyre-at-optics.rochester.edu Name: Brian McIntyre
Organization: Univ.of Rochester
Title-Subject: [Microscopy] [Filtered] Review of Student Projects
Question: Hi all-
I've just concluded a great semester of teaching EM techniques to a mixed group of undergrads and grads. They've put together some interesting projects that are posted on the web here:
} Dear Listserver, } } I have recently processed Hela cells grown on covertslips by } conventional chemical fixation( 2.5% formaldehyde plus 2.5% glutaraldehyde } for 2h and postfixation in 1.5% Osmium in 0.1M sodium cacodylate buffer, } dehydrated in a series of concentrations of ethanol over a period of 90 } minutes and then infiltrated in Epon-aradite). After sectioning, I stained } the sections with 2% uranyl acetate in distill water for 10 min and lead } citrate for 5 min. I could hardly visualize the membrane structrue, } particularly nuclear membrane, plasma membrane, Golgi apparatus and ER } membranes. However the other structures were well preserved. I am here } asking for your advice and greatly appreciate any suggestions! } } Thank you very much in advance! } } Guofeng Zhang, Ph.D } Division of Bioengineering & Physical Sciences } National Institute of Health } Bethesda, MD 20892 } Tel: 301-451-3856 } Email: zhangguo-at-mail.nih.gov } }
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 12:14:15 2004
I'm trying accquire EDS spectra of various posphors and am having difficulty. There are peaks that do not seem to match any known elements, and all of the peaks shift up and down in energy from one spectrum to the next. I'm wondering if this is a common phenomenon with phosphors, or is my system out to lunch? All ideas are welcome!
Diane Ciaburri
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 12:33:56 2004
Having been around for what seems like forever, I really did not write up some of the old literature as some of our students have suggested. But there are old reports of just this problem. The conclusion was that osmication does not adequately stabilize membranes against extraction during ethanol dehydration. The reference is a bit fuzzy, but I believe it was Dan Pease’s book ‘Histological Techniques for Electron Microscopy’, 2nd edition, 1964. It was also never clear whether it was the ethanol or the propylene oxide intermediate which was responsible for membrane extraction. We have tested this out and confirmed that, for whatever reason, in our hands we will see the loss of membranes you describe if we use the procedures as you list.
There are two solutions. First, post-fixation in uranyl acetate. Uranyl acetate will stabilize membranes. We wash 1x in buffer and 3x in filtered water after the osmication step, and then fix in 2% UA. Our monolayers and free suspensions are usually only fixed for a couple of hours, but we do leave them in UA for up to several days without noting any deleterious effect. This step usually improves stabilization of membranes in our hands.
The second solution is to dehydrate in acetone. There are two reasons, first is that acetone is not reported to extract membranes, and second, you do not need an intermediate step such as propylene oxide, but go directly into graded acetone:plastic for infiltration. This second reason saves on costs, reduces turnaround, and is a little more environmentally friendly. In fact, we have found that using acetone is, by itself, sufficient to ensure good membrane stabilization.
Our normal procedure now is to use both UA en bloc postfixation followed by acetone dehydration. Our results are usually very good, and we never lose membranes.
Hope these ideas help.
Paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 13:07:05 2004
} Can anyone suggest an application (UNIX based ideally) that I might } get started with? As a novice in this area I realize that the concept } of 3-D imaging and stereographic projection are probably two entirely } different principles. I would appreciate any references (articles, } books, websites) that will help me to understand the concepts. } Dear Steve, If you do not get sufficient info from this list, try the 3dem list:
} ADMINISTRIVIA } } To post to this list, send mail to } } 3dem-at-ucsd.edu } } To subscribe to this list, send mail to } } majordomo-at-3dem.ucsd.edu } } with the text } } subscribe 3dem } } in the body of the message (not in the Subject:). } } To unsubscribe from this list, send mail to } } majordomo-at-3dem.ucsd.edu } } with the text } } unsubscribe 3dem } } in the body of the message (not in the Subject:).
I also know that the group at the Wadsworth Center in Albany NY has prepared stereo images from 3-D reconstructions, so you might want to contact them. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 13:38:17 2004
} } I have recently processed Hela cells grown on covertslips by } } conventional chemical fixation( 2.5% formaldehyde plus 2.5% glutaraldehyde } } for 2h and postfixation in 1.5% Osmium in 0.1M sodium cacodylate buffer, } } dehydrated in a series of concentrations of ethanol over a period of 90 } } minutes and then infiltrated in Epon-aradite). After sectioning, I stained } } the sections with 2% uranyl acetate in distill water for 10 min and lead } } citrate for 5 min. I could hardly visualize the membrane structrue, } } particularly nuclear membrane, plasma membrane, Golgi apparatus and ER } } membranes. However the other structures were well preserved. I am here } } asking for your advice and greatly appreciate any suggestions!
Try increasing the time in uranyl acetate to 25 or more minutes, and decrease the time in lead citrate to 3 minutes. Unintuitively, sometimes longer times in the lead citrate causes bleaching rather than staining!
On the other hand, I've also had success with 10 minutes in each, and with 5 minutes in lead followed by 10 minutes in UA followed by another 10 minutes in lead, and many other variations. Try several approaches and see what works for you!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 14:08:00 2004
Dear Guofeng. I have worked on many cell culture preps. I normally use microwave processing however for bench processing I would recommend that you shorten all your processing steps. I would bench fix in glut and osmium for 20 mins each at room temperature for example. I use 2% glutaraldehyde on 0.1 M cacodylate buffer. I wash in 0.1M cacodylate buffer with 0.3M sucrose added. I also add 0.3 M sucrose to the post fix. The cells are very thin and not like a tissue chunk. Also, dehydration times should be very short- 1-2 mins is sufficient. in your alcohol changes. Cultured cells don't have much contrast so try using a reduced osmium fix (2% osmium tetroxide in 0.1 M cacodylate buffer containing 0.8% potassium ferricyanide). it gives really nice contrast to membranes and cytoskeleton. Also try an en bloc UA step before the dehydration. Your post staining sounds OK. Good luck JoAnn Buchanan Dear Listserver, } } I have recently processed Hela cells grown on covertslips by } conventional chemical fixation( 2.5% formaldehyde plus 2.5% glutaraldehyde } for 2h and postfixation in 1.5% Osmium in 0.1M sodium cacodylate buffer, } dehydrated in a series of concentrations of ethanol over a period of 90 } minutes and then infiltrated in Epon-aradite). After sectioning, I stained } the sections with 2% uranyl acetate in distill water for 10 min and lead } citrate for 5 min. I could hardly visualize the membrane structrue, } particularly nuclear membrane, plasma membrane, Golgi apparatus and ER } membranes. However the other structures were well preserved. I am here } asking for your advice and greatly appreciate any suggestions! } } Thank you very much in advance! } } Guofeng Zhang, Ph.D } Division of Bioengineering & Physical Sciences } National Institute of Health } Bethesda, MD 20892 } Tel: 301-451-3856 } Email: zhangguo-at-mail.nih.gov }
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 14:24:13 2004
On May 3, 2004, at 10:16 AM, Diane.Ciaburri-at-gd-ais.com wrote:
} I'm trying accquire EDS spectra of various posphors and am having } difficulty. There are peaks that do not seem to match any known } elements, } and all of the peaks shift up and down in energy from one spectrum to } the } next. I'm wondering if this is a common phenomenon with phosphors, or } is my } system out to lunch? All ideas are welcome! } Dear Diane, I assume you're looking at inorganic phosphors, such as CdS, etc., in which case there should be no problem. Just because it's a phosphor does not change the interaction with beam electrons that produces x-rays. There might be contaminants in the mix, but these would be minor--not ~10%--so there could be some very small peaks, but everything should match one or more known elements. Have you calibrated the energy spectrum? The energies should not shift--the only way that might happen is if the majority of the x-rays were inelastically scattered. If you take multiple spectra from a single area (assuming that there is not significant loss of material during data collection) neither the energies nor the intensities should vary. I suggest putting in a standard, such as Al evaporated onto a formvar film on a Cu grid, and taking spectra from locations where both the Al and Cu signals are strong. The spectra should be consistent, and you should be able to check the energy calibration and see if the resolution is OK. To see if the resolution meets spec, put in a grid with Mn evaporated on formvar and measure the FWHM of the peak. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 15:03:34 2004
Try changing your fixation protocols. I use the following fixatives to help with membrane preservation:
"Yellow Fix for EM" (based on Ito, S and Karnovsky J. J Cell. Biol Abstract 1968) 2.5% glut. 4.0% paraformaldehyde 0.02 % picric acid in 0.1M sodium cacodylate buffer
(I use 20ml of 0.2M buffer, 10ml of 10% glut, 10 ml of 16% pfa and 2 ml of a saturated sol. of picric acid--I keep a jar of it in my hood, with the crystals covered with water and draw off as I need it, adding fresh water when I'm done so that it doesn't explode.)
For a secondary fix I use 1% osmium tetroxide and 1.5% potassium ferricyanide (aqueous)made up my mixing stock solutions of 2% osmium and 3% K-ferr 1:1.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 15:06:27 2004
We have an infrared chamber-scope that I have to instruct users to turn off before they do any EDS. Otherwise, the flood of IR photons overwhelms the EDS detector and runs it up to 100% dead time. Under less overwhelming conditions, the photons are still detected and their energy adds on to the real energy of the x-ray photons and peaks get shifted upscale. Its kind of fun to watch the effect on our spectrum as we turn the light on and off and the spectral peaks shift back and forth.
I had never thought about it, but the same effect should be expected when working with cathodo-luminescent materials. And your phosphors are definitely luminescent.
I recall that some EDS detectors have their windows coated (with Al, I think) to avoid just this problem. I don't know if that is something that can be done very well to an existing detector. I don't suppose you could interpose another film in front of your detector; it would probably also absorb your x-rays.
I wonder how others would suggest dealing with this.
Warren
At 12:16 PM 5/3/2004, you wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Luci, the Microscopy Today journal, March/April 2003 issue has an article on "The Life and Death of a Tungsten Hairpin Filament". In it there is described the different failure modes for a hairpin filament which may help you.
You should be able to access this article via the following link:
Please help me. I'm really in trouble. Just burned the second filament after 3 hours. Previously, I had about 200 h. Now the current is very unstable. Before burning, is slowly going down and after that a quick increase up to 170 uA and bang. We have a Jeol 1010 (1 year old).
Many thanks, Luci _________________________________________ Lucian Barbu-Tudoran Electron Microscopy Center Faculty of Biology & Geology Babes-Bolyai University of Cluj-Napoca 5-7 Clinicilor Str., 3400 Cluj-Napoca, Romania Tel/Fax: +40 264 598700 {http://hasdeu.ubbcluj.ro/emc/lucian_barbu_tudoran.htm
__________________________________ Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs http://hotjobs.sweepstakes.yahoo.com/careermakeover
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 16:24:54 2004
We would guess the problem may be in your power supply. Your 200+ hours indicates a good vacuum and clean system. Our customers average 200+ on rebuilt filaments and slightly less on new filaments due to the special process used on rebuilt filaments.
Based on your comments, I'd check with Jeol services.
John Arnott
Disclaimer: Ladd Research is a supplier of a wide variety of microscope supplies and accessories, including new and rebuilt filaments.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
----- Original Message ----- } From: "Lucian Barbu (by way of Ask-A-Microscopist)" {lbarbu-at-hasdeu.ubbcluj.ro} To: {microscopy-at-ns.microscopy.com} Sent: Monday, May 03, 2004 9:40 AM
Bill,
Yes, the system was just calibrated last Friday, and I double-checked it again today which led me to think there must be something funny happening with the samples. Here's the answer, thanks to a variety of helpful hints I received. Apparently my detector is sensitive to visible light and the whole spectrum was being shifted to higher energies by varying amounts (depending on how much light I was generating). By switching from my thin window to my berillium window, and blocking the visible light, things make sense again.
Thanks, Bill and everyone else that responded. You guys are life savers!
Diane Ciaburri
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Monday, May 03, 2004 3:32 PM To: microscopy-at-msa.microscopy.com
Dear Diane, If you are using the modern thin-window EDS detectors sold nowadays, you should know that they are very susceptible to light and that could easily be the source of your strange signals. The old-style Be-window detectors were not light-susceptible, so if you can find one you can test the elements above Ne. For the elements below that, perhaps using a low accelerating voltage ( {5 kV) will reduce the light emission from the phosphor enough to get a reading on a thin-window detector. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {Diane.Ciaburri-at-gd-ais.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Monday, May 03, 2004 10:16 AM
Paul and Guofeng I never though for staining in UA after osmication as a "postfixation". It's hardly believe to me that UA has something to do with "fixation". In my point of view UA stained positively nucleic acids and don't interfere with membranes. When I need reduce the appearance of the ribosomes in neurone's cytoplasm, I used to remove UA staining after osmication step from procedure. In my hands, it absolutely does not affect membranes (I did both: with and without that step). I used Spurr, not Epon. I also did not see any effect of ethanol on membranes. I think, if you are using it for 10-15 min each step, it should not affect membrane's quality, otherwise nearly whole EM is pure artefact (most people still use ethanol). The basic reason, not to use acetone for dehydratation is because acetone is very strong denaturing agent (contrary to ethanol), so acetone will definitely alternate the fine structure. For this reason, people use acetone in freeze-substitution procedure at low temperature to reduce denaturing effect of acetone. Returning back to original question, I would suggest to check is OsO4 was good or not. If there is any suspicious there, you need to use fresh solution/chemicals. After osmication the tissue (even in monolayer) should turn brown. Sergey
At 12:36 PM 5/3/2004 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 18:28:33 2004
I think Diane's issue with phosphorous is visible fluorescence similar to the face of a CRT. After all, to get an image on a TV screen we shoot electrons at phosphorous coated screens and hence it fluoresces in visible light, and likely IR as well. Ideal to make the EDX detector do odd things. Chamberscopes, as Warren indicated, have the same effect.
Peter Tomic Agere Systems
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Monday, May 03, 2004 5:53 PM To: Diane.Ciaburri-at-gd-ais.com Cc: Microscopy
Dear Diane, If you are using the modern thin-window EDS detectors sold nowadays, you should know that they are very susceptible to light and that could easily be the source of your strange signals. The old-style Be-window detectors were not light-susceptible, so if you can find one you can test the elements above Ne. For the elements below that, perhaps using a low accelerating voltage ( {5 kV) will reduce the light emission from the phosphor enough to get a reading on a thin-window detector. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {Diane.Ciaburri-at-gd-ais.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Monday, May 03, 2004 10:16 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-hotmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 12:44:21 ---------------------------------------------------------------------------
Email: donaldawbrey-at-hotmail.com Name: Donald G. Awbrey HT(ASCP) QIHC
Organization: Harris Methodist Hospital
Title-Subject: [Microscopy] [Filtered] MListserver: EM scopes
Question: Dear MSA netters,
Our department is in the preliminary stages of aquiring a new electron microscope. The one in use is antiquated. I would like to have as many opinions of different scopes and their ancillary systems.
We are looking for a scope to be used for diagnostic services (such as kidney biopsies, tumors, and etc.).
Our requirements are: 120kv, flat bed camera scanner, subliminal printer, image software, dual digital/manual capabilities.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ecd10-at-psu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 14:10:00 ---------------------------------------------------------------------------
Email: ecd10-at-psu.edu Name: Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Microscopy] [Filtered] MListserver:Postdoc opening at Penn State
Question: POST-DOCTORAL POSITION in Electron Microscopy of Nanostructured Electronic Materials at The Pennsylvania State University
A postdoctoral position is available in the area of nanostructured electronic materials beginning July 1, 2004. The research project focuses on understanding interface structure and chemistry in heterostructured thin films and nanowires. Aspects of the project will be conducted under the auspices of an NSF-NIRT program on semiconducting nanowires. Through a variety of electron imaging and spectroscopy techniques we aim to quantify atomic structure and chemistry of interfaces in heterostructured materials. Most of the research will be conducted on a JEOL 2010F Field Emission TEM/STEM outfitted with an EDAX energy dispersive x-ray spectrometer (EDS), Gatan Enfina electron energy loss spectrometer (EELS), high-angle annular dark field STEM detector, and acquisition hardware and software for spectrum imaging. The ideal candidate for this position will have experience in HRTEM, STEM, EDS and EELS. The salary will be commensurate with qualifications and experience.
Please forward questions or send applications to:
Professor Elizabeth Dickey Department of Materials Science and Engineering The Pennsylvania State University 223 Materials Research Building University Park, PA 16802 USA
South Bay Technology offers 3 types of Colloidal Silica for cross sectioning of semiconductor materials - as well as other applications. The products we offer are:
All of these products are available through our local distributor in France. All of our distrinbutor contact information can be found on our website. As luck would have it, "Colloidal Silica" is also the featured product on our website today. You can go to www.southbaytech.com and find information on the colloidal silica products on the main page. Alternatively, you can enter "colloidal silica" in the product search box to bring up all references to our various colloidal silica products.
I hope this helps.
Best regards-
David
sylvain maury wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue May 4 12:41:00 2004
The clinical lab I used to work in had a Codonex di-sub printer which we rarely used. Overall, our imaging software (Gatan Micrograph ~2+years old) did not jive with the printer too well - less resolution with printed image. Not to say its a bad system. It might have been because our imaging program needed updating. Case in point, scanned images that were 'tweeked' in Adobe and printed with the Codonex came out excellent. Sara Goldston
by way of MicroscopyListserver {donaldawbrey-at-hotmail.com} wrote:
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Email: donaldawbrey-at-hotmail.com Name: Donald G. Awbrey HT(ASCP) QIHC
Organization: Harris Methodist Hospital
Title-Subject: [Microscopy] [Filtered] MListserver: EM scopes
Question: Dear MSA netters,
Our department is in the preliminary stages of aquiring a new electron microscope. The one in use is antiquated. I would like to have as many opinions of different scopes and their ancillary systems.
We are looking for a scope to be used for diagnostic services (such as kidney biopsies, tumors, and etc.).
Our requirements are: 120kv, flat bed camera scanner, subliminal printer, image software, dual digital/manual capabilities.
Many thanks to all of your helpful comments and suggestions on membrane visualization problems! I think I will get succeed in solving the problems in near future after taking your advise.
Kind Regards!
Guofeng } } Guofeng Zhang, Ph.D } Division of Bioengineering & Physical Sciences } National Institute of Health } Bethesda, MD 20892 } Tel: 301-451-3856 } Email: zhangguo-at-mail.nih.gov } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 4 17:14:38 2004
Listers, I have a researcher wanting to look at seed surface morphology. These seeds are from dried herbarium sheets. I initially had him simply treat the seeds with acetone to remove any waxes from the surface and then O-T-O-T-O them. This was not adequate as we saw what I believe to be fungal hyphae and spores plus a LOT of debris. Does anybody have a "tried and true" (and easy!) method of preparing seeds? I was thinking about doing acetolysis or 10% KOH, which I do with pollen routinely, but
wasn't sure how the seed surface would hold up. Also, I don't think the person sonicated in acetone. Might that alone be adequate? TIA.
Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
From MicroscopyL-request-at-ns.microscopy.com Tue May 4 20:01:43 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cmeyer911-at-yahoo.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 4, 2004 at 19:31:16 ---------------------------------------------------------------------------
Question: I have an old Gatan Model 607 EELS system that I have never run (I'm new to TEM). It used to be run by a TN-5500 system, that is soon to be replaced with a new EDS system. I do not want to lose EELS capability for future projects. A Gatan representative informed me the 607 system could be run with a Mac with an old operating system.
I'd like to talk (email) to any current or former 607 EELS users that would have ideas on how I could keep this capability for the future. Is my only option to keep the TN-5500 for this purpose alone? I'd appreciate it. Thanks.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jacqui.ross-at-auckland.ac.nz) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 4, 2004 at 23:18:40 ---------------------------------------------------------------------------
Email: jacqui.ross-at-auckland.ac.nz Name: Jacqui Ross
Question: We are about to buy a new inverted fluorescence microscope, probably a Nikon.
Does anyone have any experience/feedback about the Nikon Digital Sight colour camera (DS-5Mc-U1)? We are intending to buy the new cooled one and I would appreciate hearing from anyone who has used one or seen one demonstrated recently.
Here is the TABLE OF CONTENTS for the MAY 2004 issue of MICROSCOPY TODAY.
New Subscriptions via http://www.microscopy-today.com only, please. New subscriptions will close on Tuesday May 11th for this issue.
WE HAVE PURGED NON-MSA MEMBER, NON-NORTH AMERICAN, UNSUBSCRIBED INDIVIDUALS! SEVERAL HUNDRED NON-QUALIFIED SUBSCRIBERS HAVE BEEN PURGED.
Scanning Wet Specimens .Stephen W. Carmichael and Wilma L. Lingle, Mayo Clinic
Laboratory Design for High-Performance Electron Microscopy .M.A. O'Keefe, et al., LBNL; C.J.D. Heatherington, et al., Oxford; B. .Carragher, et al., Scripps; L.F. Allard, et al., ORNL
New CMEIAS Image Analysis Software for Computer-Assisted Microscopy of Microorganisms and Their Ecology .Frank B. Dazzo, Michigan State University
Making Slide Shows in Acrobat .Jerry Sedgewick, University of Minnesota
Sub-Angstrom Resolution with a Mid-Voltage TEM .M.A. O'Keefe, C.J.D. Hetherington, E. C. Nelson, LBNL, Berkeley
Viability and Versatility of the Yeast Cell .Michelle J. Henry-Stanley & Carol L. Wells, Univ. of Minnesota
New Adhesion Mechanism in Giardia: Role of the Ventrolateral Flange in the Attachment of Trophozoites to Rough and Porous Surfaces .S.L. Erlandsen, U. Minnesota; A.P. Russo, & J.N. Turner, New York .Wadsworth Center
Project VISUAL: Facilitating the Connection Between Art and Science .L.M. Strzegowski & T.P. Russell, U. of Massachusetts, Amherst
Use and Disposal of Uranyl Acetate in the Electron Microscope Laboratory: Glow in the Dark or Walk in the Park? .Randy Tindall, University of Missouri
Confocal Microscopy for Diagnostic Cytology .M.E. Boon, & L.P. Kok, U. Groningen, The Netherlands
Technical Note on the Preparation of Un-decalcified Trabecular Bone for Examination by TEM .Jeannette Taylor, Emory Univ. & Iwona Jasiuk, Georgia Inst. of Technology
The Low Voltage SEM Imaging Advantage: A Reminder .Steven S. Hurban. Endicott Interconnect Technologies, Inc. NY
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Wed May 5 16:53:09 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fremingt-at-fhcrc.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 5, 2004 at 11:24:55 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bwareham-at-utah.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 5, 2004 at 15:08:22 ---------------------------------------------------------------------------
Email: bwareham-at-utah.gov Name: Beverly Wareham
Organization: Utah Veterinary Diagnostic Lab
Title-Subject: [Microscopy] [Filtered] Fixation of animal lung tissue
Question: Hi all, Does anyone have a protocol for fixing animal lung tissue (sheep) for EM and is willing to share it? I'd really appreciate it.
Sometimes you just gotta share the good news. After trying out my share of handheld vacuum cleaners for small lab cleanup jobs and finding them all to be pretty asthmatic, I finally hit paydirt. I found something called the Euro Pro 700 watt hand vac down at the local Office Depot for about $30, complete with small hose, attachments and shoulder strap. Not only does it clean up the plastic fluff that builds up in ultramicrotomy areas, it would probably suck up the whole microtome if you turned your back. That's probably why it's also called "The Shark".
No financial interest, just a (finally) happy owner of a tiny vacuum cleaner that works!
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Wed May 5 16:54:16 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ananthpb-at-rediffmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 5, 2004 at 16:07:20 ---------------------------------------------------------------------------
I have a question for those who work in Failure Analysis of semiconductors but anyone can answer...
I'm trying to define a methodology to use colloidal silica for polishing process of microsectionned dies (Si, AsGA), for SEM observations. I'd like to know the conditions that people commonly use for their sample preparations with Syton. Any suggestions are welcome... Thanx in advance!
and thanx to David Henricks : i just received the documents about colloidal silica and SEM sample preparation. But the paper about iridium sputtering was not in the package. You must have forgotten it, but it doesn't matter. thanx again...
Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 07:28:23 2004
} hi ! } } ask the company name "logitech" (a division from struers denmark) } they are have a special equipment for polishing semiconductors ! } email info-at-logitech-us.com or www.logitech.uk.com } } best regards } } chris huebner } } Foundry Research Insitute, Kraków - Poland }
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 08:38:24 2004
I don't know if anyone else would wish to confirm my fears but I am concerned about the use of any basic vacuum cleaner for clearing up resin dust.
The problem as I see it is: 1. Vacuum cleaners do pump out small particles from their exhaust (unless they have some very fine submicron exhaust filter). 2. It's the fine stuff (below 2 or 3 microns) that remains airborne and worse still is the ideal size for inhalation. It is much more likely to penetrate deeper into the respiratory tract (hence the problem with certain sizes of dust such as asbestos). 3. Resins used for most biological purposes are not completely polymerised at 60 to 80 degrees. They are usually regarded as safe enough to handle but could well be hazardous if inhaled and retained in the lungs for long enough.
If you combine the above facts with, for instance, sawing or grinding resins (particularly epoxies like Spurr's) then you might actually be increasing your exposure to the most hazardous component of the resin dust. Generally I try to avoid sawing and only ever trim blocks with razor blades or glass knives because the pieces are big and even if you could inhale some I doubt they'd ever reach your lungs.
Some of this is assimilated information from years ago, so I can't give you a particular source or reference. I suspect that you could easily check if there is a problem by putting a 'sticky' stub near to the exhaust of your vacuum cleaner and see if it picks up much. I've never tried it because I just simply avoid fine dust now.
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu}
The International Metallographic Society is seeking a volunteer to serve as the Local Chair for the 2005 International Metallographic Contest which will be held in conjunction with the 38th Annual Convention of the IMS and the Microscopy and Microanalysis 2005 meeting in Honolulu, Hawaii in July of 2005. Interested individuals from the Honolulu area should contact the Contest Chair, Jeff Stewart, at jeff-at-metallography.com for additional information. We would like to have the position filled within the next few weeks.
Thanks,
Jeff Stewart International Metallographic Contest Chair Metallographic Laboratory Manager Stern-Leach Co. 49 Pearl Street Attleboro, MA 02703 USA Phone: 508-222-7400 extension 1329 Fax: 508-699-4030 E-mail: jeff-at-metallography.com
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 08:43:47 2004
Good points, Malcolm. I forgot to mention that this thing comes with a HEPA filter. Anyway, most of our resin fluff comes from block trimming with razor blades and glass knives.
Randy
-----Original Message----- } From: Malcolm Haswell [mailto:malcolm.haswell-at-sunderland.ac.uk] Sent: Thursday, May 06, 2004 8:42 AM To: MSA microscopy listserver Cc: Tindall, Randy D.
Micromavens,
I'm hoping to find someone with a Gatan model 626 cryostage/cryoholder who wishes to sell it or donate it to a university. Cryostation is not necessary, we've got that. Anyone out there? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 10:12:47 2004
One of faculty here will be processing lengths of rat femoral artery for light microscopy and needs to keep track of the distal vs proximal ends. She tried marking one end with India Ink, but it was washed out sometime during the dehydration/xylene steps. Any ideas? Thanks, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 10:49:32 2004
I was on a TEM project many years ago working with sheep and chicken lungs. Jeff Weidner was the PI on the project. The problem in the we had was to get the air out of the lungs before we could any of the specimen prep procedures to work well. To many air bubbles and poor infiltration.
We ended up use a standard EM procedure with each step done under low vacuum.
Mike
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From MicroscopyL-request-at-ns.microscopy.com Thu May 6 11:43:58 2004
On May 6, 2004, at 9:09 AM, Leona Cohen-Gould wrote:
} One of faculty here will be processing lengths of rat femoral artery } for light microscopy and needs to keep track of the distal vs proximal } ends. She tried marking one end with India Ink, but it was washed out } sometime during the dehydration/xylene steps. } Any ideas? } Dear Lee, Could one end be cut straight across and the other cut at an angle? I don't know how readily this could be done or whether it would be easy to distinguish the ends, but if one cannot determine the shape of the end, I'd wonder whether other morphology would be reliable. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 13:36:27 2004
Hi Beverly, We have the same experience as Mike with both lung, brain (and a few others) and most plant tissues. Before microwaving, we used the standard protocols (G, followed by Os, etc.) and did all steps in a low vacuum. You can use an evaporator using only the rough pump (Good idea to protect where the vials are set so resin doesn't get on evaporator plate_, if you don't have another vac chamber such as a vacuum oven. Now that we microwave most of our tissues, we use the cold plate and vacuum chamber for hard to penetrate tissues on all steps. It is likely that the alcohols don't need to be done in the MW however it is faster.
NOTE about pulling the vacuum. One needs to WATCH when pumping the samples. We pump on the samples and wait for the bubbles to rise. Just before they get too close to the top, we turn off the vacuum, let the bubbles settle a bit and start again. Eventually no bubbles appear. This of course is likely more crucial in the resin steps since resin all over everything is not nice! After the bubbles have subsided and don't appear in quantity, we let the vial sit in the vacuum chamber under vacuum but with the pump off, for the desired time of the step.
When using a microwave with a vacuum chamber it is important to note WHERE the gauge actually is since if it is directly off the pump, then when it pulls down to 20mmHg that is only at the pump port, not the chamber, so that must be taken into consideration. We usually use that approximate vacuum for the vac chamber on the microwave for most fixations that are done in the microwave. It is hoped, sincerely hoped, that eventually the manufacturers will build in a vacuum chamber with a cold plate in the microwave so it isn't quite so messy to set up.
Good luck, Judy Murphy
Michael Dunlap wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi Beverly, } } I was on a TEM project many years ago working with sheep and chicken } lungs. Jeff Weidner was the PI on the project. The problem in the } we had was to get the air out of the lungs before we could any of the } specimen prep procedures to work well. To many air bubbles and poor } infiltration. } } We ended up use a standard EM procedure with each step done under low vacuum. } } Mike } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (bwareham-at-utah.gov) from } } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html } } on Wednesday, May 5, 2004 at 15:08:22 } } --------------------------------------------------------------------------- } } } } Email: bwareham-at-utah.gov } } Name: Beverly Wareham } } } } Organization: Utah Veterinary Diagnostic Lab } } } } Title-Subject: [Microscopy] [Filtered] Fixation of animal lung tissue } } } } Question: Hi all, } } Does anyone have a protocol for fixing animal lung tissue (sheep) } } for EM and is willing to share it? I'd really appreciate it. } } } } Thanks, } } Beverly Wareham } } Utah Veterinary Diagnostic Laboratory } } 435-797-1799 } } bwareham-at-utah.gov } } } } --------------------------------------------------------------------------- } } -- } =================================================================================== } Michael Dunlap } office (530) 752-0284 } University of California lab (530) 752-5489 } Chemical Engineering & Material Science Fax (530) 752-9554 } 110A Kemper Hall mrdunlap-at-ucdavis.edu } One Shields Ave. } http://www.chms.ucdavis.edu/ } Davis CA, 95616 } ===================================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 14:26:10 2004
When you use India Ink it needs to be fixed to the tissue's surface you are labeling with a picric acid solution such as Bouin's fixative.
Make the ink mark with the opposite end of a swab and then switch to the cotton tipped end of the swab. Dipp into the Bouin's and dab the area marked with the India Ink. Blot slightly dry with a Kim Wipe and put back into formalin. It will be permanently fixed in place and can be processed and embedded in paraffin.
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 14:53:29 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sbs38-at-email.byu.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, May 6, 2004 at 10:14:58 ---------------------------------------------------------------------------
Question: I am collecting protocols for viewing HSV-1 through negative staining, and I was hoping you could send what you feel is the best protocol for viewing membrane and capsid integrity. Next, I need to know the best stain and method for viewing empty or full capsids. Thank you for your reply
We here at the University of New Brunswick have a Jeol JXA-733 Superprobe that has served us faithfully for many years. Sadly we have had a catastrophic failure of the oil-cooled objective lens/pole piece assembly, and so require a replacement. If you have working one and would be willing to part with it, please contact us.
Thanks for your time,
Steven Cogswell, P.Eng. UNB Microscopy and Microanalysis Facility Fredericton, New Brunswick Canada Phone (506) 453-4887
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 18:32:09 2004
You could try OsO4. Its guaranteed to darken on the specimen and it won't wash out. The only concern is how useful that end will be once its been treated with Osmium. I know Osmium is used to fix fatty specimens in order to give them more contrast. I would use a 1% or maybe a 2% solution. It usually takes effect within a few minutes, so you could maybe dip the distal or promixal portion in the solution. Good luck. Sara Goldston "Bentley, Karen" {Karen_Jensen-at-URMC.Rochester.edu} wrote:
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Hi Leona:
When you use India Ink it needs to be fixed to the tissue's surface you are labeling with a picric acid solution such as Bouin's fixative.
Make the ink mark with the opposite end of a swab and then switch to the cotton tipped end of the swab. Dipp into the Bouin's and dab the area marked with the India Ink. Blot slightly dry with a Kim Wipe and put back into formalin. It will be permanently fixed in place and can be processed and embedded in paraffin.
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
__________________________________ Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs http://hotjobs.sweepstakes.yahoo.com/careermakeover
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 21:04:00 2004
I recently received the parts to a Zeiss Axiotron microscope, which is no longer supported by the manufacturer. Does anyone have a manual or schematics for this unit--I hate to waste the box of very high quality lenses that came with it, and our engineering department is very interested in getting the unit assembled and working.
Anyone that can help me out with this, please contact me.
thanks in advance
Steve Barlow -- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
From MicroscopyL-request-at-ns.microscopy.com Fri May 7 14:34:41 2004
I'm hoping the microscopy community can help me. I had planned on taking a course on imaging analysis and photomicrography at McCrone Research but it was canceled (I was the only student.)
I have a real need to improve my imaging and image measurement skill. I wish to do particle sizing on images captured via the SEM/TEM/light microscope.
Does anyone know of short courses and such for the tyro? I would like take one as soon as possible.
Thanks in advance!!!
Frank Karl 330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Fri May 7 18:15:57 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (e.gagnepain-at-wanadoo.fr) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, May 7, 2004 at 10:04:31 ---------------------------------------------------------------------------
Email: e.gagnepain-at-wanadoo.fr Name: Gagnepain
Organization: ULP of Strasbourg
Education: Graduate College
Location: Strasbourg, France
Question: I have some samples of iron-aluminide (FeAL)and the thickness is 2 millimeter. How to prepare thin layer of FeAl for an observation with TEM (Transmission Electron Microscope) without introduce defaults?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cryanez-at-criba.edu.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 7, 2004 at 14:09:42 ---------------------------------------------------------------------------
Question: I need to stain a polymer sample with RuO4, by oxidation of RuCl3 with sodium hypochlorite. I don¥t know the appropiate reactives concentrations. Could you help me about this procedure?. Thanks in advance
Julia Y·Òez
Ing. MarÌa Julia Y·Òez TE: 54-291-486-1666 int.: 362 Lab.Microscopia ElectrÛnica FAX: 54-291-486-1527 CRIBABB e-mail: cryanez-at-criba.edu.ar Camino La Carrindanga Km 7 (8000) BahÌa Blanca
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I have never prepared specimens from this material, so I'm not familiar with its properties.
Is it electrically conductive?
If so, to begin with I would spark errode with a trepanning tool to get 3 mm dia. discs. Then mechanically grind and polish, preferably with a dimple grinder, to ~100 um. Finally, electropolish, preferably with a jet polisher.
Hirsch, Howie, Nicholson, Pashley and Whelan suggest, for Fe-Al alloys acetic acid (133 cm3), water (7 cm3), chromic acid (25 g) with a stainless steel cathode and rinses in acetic acid/methanol. Make sure it remains cool ( {30 C). Voltage in the 25 V to 30 V range.
Possible problems:
1. Different phases may thin at different rates during electropolishing - you'll need to change the polishing solution.
2. If it is very soft, the mechanical grinding and polishing may create a deep damage layer - stop mechanical polishing sooner and use electropolish earlier.
3. If there is stress in the material, thinning may cause a release of stress and damage.
4. Safety officers
If you can't get electropolishing to work, ion milling should. -- Larry Stoter PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail will automatically be deleted. :-) PS - I'm an employee of JEOL UK.
From MicroscopyL-request-at-ns.microscopy.com Sat May 8 06:25:30 2004
This humble amateur histologist always used "alcoholic Bouin" ("Duboscq-Brazil Fluid") combined with gentle vacuum applied the way Judy Murphy describes it, to fix small pieces of rabbit lung...
Try google with "alcoholic bouin" as the search string.
"alcoholic Bouin" (aka:"Duboscq-Brazil Fluid")
Stock Alcoholic Bouin's Solution: 80% ETOH 750.0 ml formaldehyde 300.0 ml Picric acid 5.0 gm
Mix solution well, stable 1 year.
Working Solution: Stock solution 70.0 ml 14.0 ml Acetic acid 5.0 ml 1.0ml Add the acetic acid right before use.
Y.
-----Original Message----- } From: Judy Murphy [mailto:jmurphy-at-deltacollege.org] Sent: donderdag 6 mei 2004 20:22 To: Michael Dunlap Cc: Microscopy-at-msa.microscopy.com; bwareham-at-utah.gov
briant
the question really is what is the source of specimen? are you looking for clinical isolates or at culture material. if culture material, has it been purified in any way. these things may all affect the preparation and ultimate quality.
the first issue you have to address is grids. you will need grids with some form of plastic film, be it collodion, formvar or parlodian. if you are not set up to make grids, they can be obtained from most EM supply houses. for short term situations, if you are not going to need a lot of grids and have no one trained in their preparation, buying them will be best for your use. you may also wish to carbon coat the grids to stabilize the film. again, carbon coated plastic coated grids may be purchased. if cost is a problem, i could probably manage to liberate 50-100 grids.
carbon coated grids introduce several problems, mainly due to the fact that they are quite hydrophobic. however, they do become more hydrophilic over time. also, you can treat the grids by glow discharge (within 2-3 weeks of use) or treat them with alcian blue. usually clinical material has so much serum or fecal protein present that if you leave the sample on the grid for several minutes you overcome all problems of hydrophobicity. we view 2500-3000 clinical samples, and over 4000 total negative stain preparations/ year. at the risk of being labeled a reprobate and cretin, we usually do not carbon stabilize the grids used to examine most samples. the added stability is not enough benefit for clinical material. if it is a sample of importance we may use carbon coated grids. if so, we use alcian blue, or old grids. either way, with our concentration methods, hydrophobicity is not a problem.
you may wish to consider pretreatment of the sample to stabilize the virion. we either add glutaraldehyde to a concentration of 0.1%, or formaldehyde to a concentration of 1-2%. this allows us to neutralize infectivity, maintain antigenicity and reactivity for IEM, and stabilizes structure against stresses which may arise during preparation. the samples need to be vortexed very well immediately to limit clumping which may be associated with fixative mediated crosslinking of virions. because of the shorter chain size and mono aldehyde nature, some argue that formaldehyde is less likely to result in crosslinking, and is better than glutaraldehyde. personally, we have never seen any difference between the two.
you do not mention mounting the sample on the grid. i assume you have some method of preference. if not, please ask, we all have our favourite methods and are more than willing to tell you them.
as far as staining, most viral electron microscopists have their own preferences. that does not make any one method 'best', however. we use several stains routinely, and do a panel 14 different negative stains whenever we start a new project to confirm the best set of conditions. that said, the two major staining methods use tungsten or uranyl salts.
the most common tungsten salt is 1.5 to 2% phosphotungstic acid (PTA). to eqalize the different methods, i have adjusted the concentrations of stains in my labs to maintain an equal amount of heavy metal atoms available in the staining solution. therefore, we use 2.5mM PTA. this works out to be approximately 1.6%. the pH is adjusted to 7.0. the best method for adjusting pH with this stain is with NaOH. early studies by Horne and his associates implicated the loss of membrane integrity with pH adjustment by KOH. if you were working with paramyxo or myxo, and wanted to release the nucleocapsid, this would be a good method. for the greater part you do not wish to disrupt the membranes. with most herpes preparations, you will already see a lot of naked complete and empty nucleocapsids as it is. we also add 25 micrograms/ml bacitracin as a surfactant. this helps with stain spreadability and penetration. other small molecules are available for use. the most common is probably BSA. again, the addition of fixative will also stabilize the virions against loss of membranes due to stain characteristics.
we use two different versions of uranyl stains, uranyl acetate(UA) and uranyl sulfate (US). both of these must be used at low pH. in fact, if you try to adjust the pH to around 4.0 you start to get problems keeping material in solution - especially if your pH meter is not as accurate as you think. as a rule, we use the material at pH of about 3.5. again, most protocols call for 2% solutions. adjusting to molar concentrations and maintaining equivalent heavy metal atoms, we use a 60mM solution. this represents a 2.5% UA or 2.2% US stain. if you must work with higher pH, it has been suggested that 1% UA can be adjusted to pH values as high as 7.0 by the addition of 0.1% ammonium acetate and 0.5% EDTA (disodium salt). i have not tried this, so i do not know how well it works. rumour is that it is quite grainy and sublimates badly.
each stain system has its own properties. many feel particles degrade in PTA but not in UA. PTA has a higher anhydrous density, so you see more contrast, which could be bad or good. UA tends to be more granular when imaging, while PTA is low granularity. here the issue is purely what you think would 'look best'. UA/US tends to sublimate in the beam, leading to a perceived column contamination problem, while PTA is very stable, leading some to argue that the column is less contaminated by vaporizing stain.
the best suggestion - try both PTA and a uranyl stain. i would also refer you to a recent review: Hazelton, P.R. and Gelderblom, H.R. (2003) The use of the electron microscope for rapid diagnosis of viral agents in emergent situations. Emerging Infectious Diseases 9:294-303. it will give some additional methods and general information. yes, i'm human, it is one of my articles, but i think it's good, anyway.
hope these thoughts help. let me know if you want specific protocols. we have them all in electronic file format, albeit WordPerfect files, so they can be sent easily.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Sun May 9 19:06:16 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hunny_pot-at-optusnet.com.au) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May 9, 2004 at 21:45:15 ---------------------------------------------------------------------------
Email: hunny_pot-at-optusnet.com.au Name: Linda
Organization: The University of Sydney
Education: Undergraduate College
Location: Australia
Question: Hello:
I am doing research paper on quanification of shrinkage due to fixation. There are some questions that I like answers to:
1. what are some of the methods used to measure shrinkage?
2. how the effects of foxation can be differentiated from other possible sources of shrinkage?
3. the practices and clincal signifance of quantifying shrinkage due to fixation.
I have been looking around, but I can't find my answers. Could you please help me? Thanks
We are looking for a second hand Gatan 666 PEELS, to fit on VG HB5 STEM. Perheps are there somme which are sleeping in cupboards ! It doesn't matter if modifications are necessary. Preferently, for administrative raions, we would prefer a europeen source, but all propositions are welcome.
Answer directly to my collegue, at Daniel.Spor-at-ipcms.u-strasbg.fr
Many Thanks.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
What type of silver charged epoxy will be used to make embedding of powder as standards for EDS microanalysis. If one looks at Epotek's catalog, there are so much type of epoxys, with very different properties, shelf live and price. So what is the best compromise. If someone has an advice.
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
A great deal of research was done on these questions in the late 1940's-1960's and you won't find it on the internet. For starters, get a hold of "Principles of Biological Microtechnique" by John R. Baker, published in 1958. He discusses various fixatives, their penetration rates, how they do or do not promote shrinkage (in the fix or in subsequent processing), etc. I will try to dig up some references to specific papers later today.
Geoff
by way of Ask-A-Microscopist wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 09:43:36 2004
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Julia,
Prepare only the amount of stain that you will use at that moment. For 1 ml of RuO4 stain, use 0.02 g RuCl(subscript: 3 )(superscript: .) nH (subscript: 2)O [14898-67-0] and 1 ml of NaOCl [7681-52-9].
I recommend that you purchase fresh hypochlorite every few months since the concentration of hypochlorite in solution does diminish with time. I discourage using bleach for the same reason, i.e. one is never sure how long the bottle of bleach has been on the shelf. Keep the RuCl(subscript: 3) hydrate in a nitrogen desiccator when not in use.
The original reference on the use of the in-situ preparation of RuO4 is: -- Montezinos, D., Well, B.G. and Burns, J.L., J. Polymer Sci.: Polym. Lett. Ed., 1985, 23, 421.
Another important reference pertains to vapor-staining with RuO4 (using RuO4 crystals, which are very difficult to find): -- Sano, H., Usame, T. and Nakagawa, H., Polymer, 1986 27, 1497.
I hybridized the methods of Sano and Montezinos, i.e. using vapor staining above the in-situ preparation of RuO4, and invariably get outstanding results: -- Brown, G. M. and Butler, J. H., Polymer, 1997, 38 (15), 3937.
Although the Montezinos and Sano papers are key, their practicality is limited. Montezinos recommends staining in the solution using precursors that are dirt cheap and readily available. Sano convinced me, however, that solution staining is not a good practice because it produces severe and uncontrolled oxidation of the outer surface of the sample. Sano's vapor-staining method is beautiful but the solid RuO4 crystals he uses are virtually impossible to find in the United States and are very expensive. I simply combined the two methods and worked out staining durations. Refer to the appendix of my paper for details including safe use, handling and disposal as well as sample preparation from beginning to end and staining durations for some polymer systems.
Happy staining,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
cryanez-at-criba.edu.ar (by way of To: microscopy-at-ns.microscopy.com MicroscopyListserver) cc: Subject: [Microscopy] viaWWW: stain a polymer
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cryanez-at-criba.edu.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 7, 2004 at 14:09:42 ---------------------------------------------------------------------------
Question: I need to stain a polymer sample with RuO4, by oxidation of RuCl3 with sodium hypochlorite. I don¥t know the appropiate reactives concentrations. Could you help me about this procedure?. Thanks in advance
Julia Y·Òez
Ing. MarÌa Julia Y·Òez TE: 54-291-486-1666 int.: 362 Lab.Microscopia ElectrÛnica FAX: 54-291-486-1527 CRIBABB e-mail: cryanez-at-criba.edu.ar Camino La Carrindanga Km 7 (8000) BahÌa Blanca
You may be interested in a paper I published a few years ago in which I measured the size distribution of cells in suspension after different fixation conditions. Develop. Biol 23:36-61 (1970)
Joel
} } } ---------------------------------------------------------------------- } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (hunny_pot-at-optusnet.com.au) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May } 9, 2004 at 21:45:15 } ---------------------------------------------------------------------- } ----- } } Email: hunny_pot-at-optusnet.com.au } Name: Linda } } Organization: The University of Sydney } } Education: Undergraduate College } } Location: Australia } } Question: Hello: } } I am doing research paper on quanification of shrinkage due to } fixation. There are some questions that I like answers to: } } 1. what are some of the methods used to measure shrinkage? } } 2. how the effects of foxation can be differentiated from other } possible sources of shrinkage? } } 3. the practices and clincal signifance of quantifying shrinkage due } to fixation. } } I have been looking around, but I can't find my answers. Could you } please help me? Thanks } } ---------------------------------------------------------------------- } ----- }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 09:49:51 2004
What is the material of interest? You need to say. Obviously it can't be silver but you may want to check to see if you have silver spectrally overlapping the material or element of interest. Is your concern primarily sample charging therefore you want a conductive epoxy?
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] Sent: Monday, May 10, 2004 9:09 AM To: Microscopy Society of America
Hi
What type of silver charged epoxy will be used to make embedding of powder as standards for EDS microanalysis. If one looks at Epotek's catalog, there are so much type of epoxys, with very different properties, shelf live and price. So what is the best compromise. If someone has an advice.
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Jacques,
I spent a considerable amount of time working on conductive embedding media. See my abstract in the 2004 M&M Proceedings.
The problem with virtually all commercial electrically conductive embedding media that I have seen is that they offer only macroscopic conductivity. This means that expanses of unfilled, non-conducting and highly charging epoxy will be found throughout the sample. We need microscopic conductivity in which the embedding medium is highly conductive and does not contain non-conductive domains of epoxy. For expoxy-embedding applications, I recommend compounding (mixing) carbon black filler into the resin of choice (without catalyst or accelerator) in a mortar and pestle. Use equal parts by weight of the filler powder and epoxy. At the last minute before embedding, mix the accelerator into the resin / carbon black compound as quickly and thoroughly as possible. Embed the sample in this thick paste.
If well mixed, the embedding medium will not charge at all. Also, since only carbon and oxygen are present in any appreciable concentration, you will not have problems with interference lines from metal filler particles, i.e. silver, etc.
Have fun.
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Faerber Jacques {Jacques.Faerber-at-ipcms.u- To: Microscopy Society of America {Microscopy-at-MSA.Microscopy.Com} strasbg.fr} cc: Subject: [Microscopy] EDS standards embedding
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi
What type of silver charged epoxy will be used to make embedding of powder as standards for EDS microanalysis. If one looks at Epotek's catalog, there are so much type of epoxys, with very different properties, shelf live and price. So what is the best compromise. If someone has an advice.
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Our standards from Tsousimis came prepared with regular epoxy and a coating of carbon. We have used the same procedure for our homemade standards. Many of the standard materials are not conductive anyway, so the conductivity has to come from a coating. Of course a "conductive" epoxy should not hurt.
I presume you will be not be collecting spectra in variable pressure mode. The scattering of the beam from the residual atmosphere would produce a signal from the silver or nickel in your epoxy and would complicate things.
I also advise you to prepare the sample and standards the same. I thought I was competent at EDS standards and realized fairly recently the difference that can be caused by different thickness of C coatings (or lack thereof) between standards and unknowns.
Warren Straszheim
At 08:08 AM 5/10/2004, you wrote:
} Hi } } What type of silver charged epoxy will be used to make embedding of powder } as standards for EDS microanalysis. If one looks at Epotek's catalog, } there are so much type of epoxys, with very different properties, shelf } live and price. So what is the best compromise. If someone has an advice. } } Many thanks } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess ; BP43 } 67034 Strasbourg CEDEX 2 } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)3 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 10:28:32 2004
The Philips EM400 installations I know of all utilized a Haskris chiller. We're looking to buy a new chiller and they are considerably cheaper from Neslab. Would anyone recommend against going with Neslab for any reason?
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 10:56:54 2004
Does anyone know of an archived discussion or a web site anywhere that describes adapting a C-mount for a digital camera to the old Philips EM400 TEM? Perhaps by custom machining a cover blank for the 35mm camera mounting location?
We recently purchased a Pax-it 2.1 Mpixel digital camera to use with a new inverted metallurgical LM, but after receiving the camera, funding for the microscope didn't materialize, so we're thinking of using the Pax-it on our EM400.
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 13:52:13 2004
The only thing I don't particularly care for in our (?8-year-old) Neslab chiller is the fact that the water reservoir is pretty well sealed up, like a miniature 45-gallon drum on its side, so you can't see what's going on in there. The Haskris chillers always used to have a water tank with a lid, making for much easier service and monitoring of water condition. Other than that, both types are pretty well just pumps directly driven by electric motors with some miscellaneous tubing and controls. Both brands used to use the same pumps (made by Procon of Tennessee), and probably still do. Our Neslab has been pretty trouble-free; it's gone through one motor so far and at least two pumps, but, as they say, your mileage may vary.
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: Eric Anderson [mailto:andersone1-at-southernct.edu] Sent: Monday, May 10, 2004 1:03 PM To: MSA listserver
Ooh, one more question:
The Philips EM400 installations I know of all utilized a Haskris chiller. We're looking to buy a new chiller and they are considerably cheaper from Neslab. Would anyone recommend against going with Neslab for any reason?
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 15:32:29 2004
We have been using a Neslab chiller since the beginning of time (I can't find the original records). It runs without much complaint, most of the time. We have replaced the motor (just an electric motor available from Grainger's) once, replaced the pump once, and had the old pump rebuilt by Procon once. We have replaced the shear pin ("nylon coupling")in the pump head a number of times (maybe as much as once a year), and recharged the compressor once in the last 6 years.
I've heard that the Haskris is a "better" chiller...but the Neslab seems to do the job just fine. Their service support--by phone--is wonderful, and they're in southern NH if you need to send something to them.
I highly recommend having a supply of shear pins available, and definitely have them teach you how to change them if you never have done this before--it's a little tricky the first time.
No interest in Neslab other than long experience as a user...
Deborah H. Gaynor, Ph.D. Scanning Electron Microscopist Analytical Services, Inc. PO Box 515 Williston, VT 05495 802-878-5138 x21
-----Original Message----- } From: Thomas, Frank [mailto:FThomas-at-nrcan.gc.ca] Sent: Monday, May 10, 2004 2:58 PM To: 'Eric Anderson'; MSA listserver
Eric -
The only thing I don't particularly care for in our (?8-year-old) Neslab chiller is the fact that the water reservoir is pretty well sealed up, like a miniature 45-gallon drum on its side, so you can't see what's going on in there. The Haskris chillers always used to have a water tank with a lid, making for much easier service and monitoring of water condition. Other than that, both types are pretty well just pumps directly driven by electric motors with some miscellaneous tubing and controls. Both brands used to use the same pumps (made by Procon of Tennessee), and probably still do. Our Neslab has been pretty trouble-free; it's gone through one motor so far and at least two pumps, but, as they say, your mileage may vary.
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: Eric Anderson [mailto:andersone1-at-southernct.edu] Sent: Monday, May 10, 2004 1:03 PM To: MSA listserver
Ooh, one more question:
The Philips EM400 installations I know of all utilized a Haskris chiller. We're looking to buy a new chiller and they are considerably cheaper from Neslab. Would anyone recommend against going with Neslab for any reason?
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 17:03:35 2004
Has anyone tried any of the anti-GFP antibodies on the market for western blotting? Many seem to be cheaper than Clontech for the same amount but we don't want to buy an untested antibody. Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 18:18:11 2004
What are you trying to look at with your EDS? What kind of EDS is it?
The "normal" calibration of EDS is done at Al K alpha (1.486KeV) and Cu K alpha (8.040KeV). This sets resolution of the EDS according to these two peaks. The other checks I do are done at C, O and F and at N & O. And finally, FWMH at Mn.
For these measurements, I use X-Checker and X-Checker Extra. For C, O and F, I use an Al stub with a Carbon sticky tab with Teflon tape about 50% covering the stub and the other 50% split between sticky tab and bare Aluminum pin stub. These readings do not calibrate the system. They produce resolution figures for ability to resolve light elements. You can do similar analysis for heavier elements with other stub configurations.
The rationale for light element work is to see if the EDS system, over time, drifts in the wrong direction. Then I would know that something is going bad, either window problems or EDS system problems. You will see this right away at low Z.
If you are looking for ISO standards, I'm not sure how that would work. Cu is Cu and Al is Al. The Cu and Al peaks are used. If any foreign material was present, it is simply ignored. The EDAX calibration routine only looks at the Al and Cu peaks.
gary g.
At 06:08 AM 5/10/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon May 10 18:27:40 2004
I have been running my Haskris, on my EM410, virtually continuously, since November 1982. I have replaced no parts. I have another Haskris on my 1999 Hitachi S3500-N that has an intermittent algae problem, and has also needed the starter capacitor on the motor replaced.
Rick A. Harris, exDirector Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://microscopy.mcb.ucdavis.edu raharris-at-ucdavis.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 11 02:06:04 2004
The question comes for a job on thin film of materials such Sr2TiMoO4 and CoFe2O4 grown by laser ablation. The goal is to control the stochioametric variation from the films, in comparaison with the target material, which we have as a powder, or as sintered target. So I need to embed the powder, or pieces of the sintered bulk, to make a polished surface of it. I reed in a paper from Geller in the NIST bull. mentionned by someone on the list about the use of silver charged epoxy. As I use some by the way for fixing sample for spark machining, I was in question about what kind could do the job for my standards.
As I have understand the electrical conductivity in such materials will be done by tunneling effect between the silver grain, through the epoxy. So it is not so macroscopic. I'll try the methode with carbon black. Seems to be interesting and much cheaper than all the commercial silver epoxys. And with carbon, the problem of siver L lignes signal will be avoid.
These standards are used as references for thin films analysis, using Stratagem thin film software.
Some other suggestions ? Many thanks.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Jacques; } } What is the material of interest? You need to say. Obviously it can't be silver but you may want to check to see if you have silver spectrally overlapping the material or element of interest. Is your concern primarily sample charging therefore you want a conductive epoxy? } } Regards, } } Peter Tomic } Agere Systems } } -----Original Message----- } } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] } Sent: Monday, May 10, 2004 9:09 AM } To: Microscopy Society of America } Subject: [Microscopy] EDS standards embedding } } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi } } What type of silver charged epoxy will be used to make embedding of powder } as standards for EDS microanalysis. If one looks at Epotek's catalog, } there are so much type of epoxys, with very different properties, shelf } live and price. So what is the best compromise. If someone has an advice. } } Many thanks } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess ; BP43 } 67034 Strasbourg CEDEX 2 } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)3 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 11 03:24:48 2004
this may be drifting off topic a bit, but we used to have algae problems with our water cooler until I covered the translucent plastic hose with aluminium foil. There doesn't seem to have been a problem for over a year and there is little need to clean out the system these days.
I know that you can get black pressure hoses as an alternative but you can't buy them from your local supermarket for ~ 1 UK pound and fit them in a few minutes while the system is running.
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK
e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: "Rick A. Harris" {raharris-at-ucdavis.edu}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mivalmar-at-yahoo.es) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 11, 2004 at 15:10:15 ---------------------------------------------------------------------------
Email: mivalmar-at-yahoo.es Name: Miguel V.Serrano
Question: I want to ask if someone has tryed to do an immunohistochemistry of PPAR alpha (peroxisome proliferator receptor alpha) in rat liver or another tissue ??. Thanks.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donovanl-at-ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 11, 2004 at 12:50:14 ---------------------------------------------------------------------------
Email: donovanl-at-ufl.edu Name: Laura Donovan
Organization: University of Florida College of Veterinary Medicine
I'm sorry that this is a bit of a deviation, but, knowing how diverse this list is, somebody may be able to help:-
I have a Stanton-Redcroft Thermal Analyser, an STA-1600, the company in its last years was acquired by Polymer Laboratories and then by Rheometrics, and I'm interested in exploring the possibility of hooking analytical instruments, maybe NDIR or even mass-spec, to the gas outlet line, to analyse the evolved gases (what was referred to some time ago, so nicely, by Stanton-Redcroft as "hyphenated techniques").
Does anyone have such a setup?
Presumably one would use a heated PTFE interface tube, but the furnace internal volume is fairly large, and not necessarily efficiently swept by the 50 ml/min or so of purge gas, and what I'm wondering about is the time resolution of such EGA in this instrument.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 04:52:33 2004
} The "normal" calibration of EDS is done at Al } K alpha (1.486KeV) and Cu K alpha (8.040KeV). } ...
Out of curiosty, why would a calibration use these 2 peaks rather than CuK and CuL?? Understandably, the L-line would be a convolution of the L-family, but I would think the Al K line is in a more difficult location relative to the slope in the continuum(???) Is there any preference with respect to EDX window type? (e.g., Be v UTW)
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 05:04:38 2004
We are looking for the RIGHT answers. Addressing different institutions across the globe appear to be a very difficult. We have been sent into circles for months without one clear cut answer.
Our EM technician has a HND in Environmental Science. He has been in the EM Unit for just over a year and is slowly getting the hang of things. We would like to promote him up the ranks. He has got the right aptitude, has matured and is responsible. Nice to find people like that still.
For him to be promotable, the University require at least a first degree (Senior Technician) and Masters (Chief Technician). Without going through the rest of the gory details, he need further study. It is the policy of the University of Botswana to send citizens abroad rather than to study locally to enrich the nation academically.
The ranking of preference is 1) South Africa, 2) Australia, 3) USA, 4) UK.
He would prefer to do a EM focused masters. If he can do a pure Electron Microscopy Masters through course and lab work the better. The preference will be in the field of Material Science if he can not do a masters in "electron microscopy".
1) For him to be send by the University for a first degree and a masters consecutively, he must find at least partial sponsorship/bursary. 2) Who must he contact. (Please one name only in your institution, not a string of people!) 3) Can he get Credit for his current qualification? 4) Academic year start, requirements, and duration of study.
We are keeping our fingers crossed. Thanks
Since my UB mail address is not reliable please send a copy of all urgent mail to coetzeesh-at-yahoo.co.uk
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
FWIW, we have a Leybold quadrapole RGA which we often connect to our vacuum furnace to monitor evolved gasses. Since the RGA operates at much higher vacuum than the furnace, the interface is made using conflat flanged s-steel fittings and a quality leak valve.
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, May 12, 2004 1:45 AM To: Microscopy-at-sparc5.microscopy.com
I'm sorry that this is a bit of a deviation, but, knowing how diverse this list is, somebody may be able to help:-
I have a Stanton-Redcroft Thermal Analyser, an STA-1600, the company in its last years was acquired by Polymer Laboratories and then by Rheometrics, and I'm interested in exploring the possibility of hooking analytical instruments, maybe NDIR or even mass-spec, to the gas outlet line, to analyse the evolved gases (what was referred to some time ago, so nicely, by Stanton-Redcroft as "hyphenated techniques").
Does anyone have such a setup?
Presumably one would use a heated PTFE interface tube, but the furnace internal volume is fairly large, and not necessarily efficiently swept by the 50 ml/min or so of purge gas, and what I'm wondering about is the time resolution of such EGA in this instrument.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 07:42:46 2004
shAf, I've thought that for a long time but have yet to find a software-based calibration that can use the 2 Cu peaks. Maybe the vendors can speak to this.
As to Gary's question of why one needs standards, I have a hunch they're not looking for EDS calibration standards (a piece of Cu tape on an Al stub works just fine) but quantitative analysis standards for high quality quant work. Despite the software advances in standardless analysis, everyone I've talked to who really knows x-ray says that standardless quant just doesn't have the accuracy or precision of standards-based analysis.
Ken Converse owner Quality Images third party SEM service Delta, PA
michael shaffer wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed May 12 08:48:21 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sp-bf-at-physik.upb.de) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 12, 2004 at 05:24:26 ---------------------------------------------------------------------------
Our physics group got a Hitachi H-600 a few years ago from biology group. We repaired it and there was no problem. But now the wire from the linkage of the specimen chamber has broken. It jumped out from the guide rolls and now we cannot move the specimen in one direction. "Nissei Sanyo Europe",the Hitachi Service Center here, demands 3500Ä for repair of this silly wire. I hope anyone can help me!!! Is there a plan (blueprint?) how to thread the wire through the rolls, so that we could repair it ourself, not perfect but in this way, that moving of the specimen would be possible?
I am trying to label the GFP in yeast on either Lowicryl sections or Tokuyashu cryosections. So far, we have had rather poor labeling/high background. I would appreciate any recommendations on a particular commercially available antibody and the fixation protocol.
Also, we would need to label a biotin-tagged protein. We were trying Streptavidin - gold, but to little success. Is there a good anti-biotin antibody (or, even better, anti-biotin/gold conjugate) people have used for this purpose before?
Any recommendations would be highly appreciated.
Michael Jarnik, EM Facility Fox Chase Cancer Center Philadelphia, PA
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 10:50:57 2004
Just guessing, but I would suppose the energy calibration is a rather simplistic algorithm. I am pretty sure that is much easier to work with a symmetric K peak and pick off the centroid than it is to deconvolute the L peaks to find the energy of the alpha line.
Presumably the effect of the background slope and curvature is negligible compared to the peak intensity if all they have to do is get a peak energy.
Warren
At 04:57 AM 5/12/2004, "michael shaffer" wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 12:36:06 2004
i', sorry, but your second question is not clear to me. are you simply trying to label your protein with gold, or is this a detection step where you have already used the antibody to react with its antigen and now want to see if where the antigen is? also, do you have any antibody which is not tagged with biotin and that you could use for tagging with either nanogold or directly onto larger gold labels? finally, could you do an indirect reaction using gold tagged antibody to the host species antibody you are using, eg, say the biotin-antibody is an IgG species, raised in mouse, could you come back with colloidal gold:anti mouse IgG?
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-392
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 12:38:32 2004
On May 12, 2004, at 2:57 AM, michael shaffer wrote:
} Out of curiosty, why would a calibration use these } 2 peaks rather than CuK and CuL?? Understandably, } the L-line would be a convolution of the L-family, } but I would think the Al K line is in a more difficult } location relative to the slope in the continuum(???) } Is there any preference with respect to EDX window } type? (e.g., Be v UTW) } } I've thought that for a long time but have yet to find a } software-based calibration that can use the 2 Cu peaks. Maybe the } vendors can speak to this.
Dear Mike and Ken, Since the Al k-line is at 1.4 keV and the Cu l-line(s) is at ~1 keV, the Al is more readily separated from the continuum. I don't know of a difference between Be and UTW calibrations, since it is the response of the crystal that is being measured, but, perhaps there is an advantage to using a lower-energy line with the UTW. The TN2000 had software that could use any two lines. One simply entered the energies and ran the calibration routine. I guess someone must have "improved" the system by entering the line energies for us. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 12:54:20 2004
Sorry, I would try to make it clearer. We need to localize a viral protein which has been biotin-tagged and expressed in cell culture. I expected it should be no problem to label it with Streptavidin conjugate, but it is not that easy, it seems (even according to some other people). So, as we do not have a good antibody to the viral protein, I am considering an indirect labeling with anti-biotin antibody and a secondary conjugate. There are tons of commercially available anti-biotin antibodies, but certainly not all of them will work with EM protocols - so I was interested whether somebody has experience with some of these.
Thanks,
Michael
paul r hazelton wrote:
} michael } } i', sorry, but your second question is not clear to me. are you simply } trying to label your protein with gold, or is this a detection step } where you have already used the antibody to react with its antigen and } now want to see if where the antigen is? also, do you have any antibody } which is not tagged with biotin and that you could use for tagging with } either nanogold or directly onto larger gold labels? finally, could you } do an indirect reaction using gold tagged antibody to the host species } antibody you are using, eg, say the biotin-antibody is an IgG species, } raised in mouse, could you come back with colloidal gold:anti mouse IgG? } } paul } } Paul R. Hazelton, PhD } Electron Microscope Unit } University of Manitoba } Department of Medical Microbiology } 531 Basic Medical Sciences Building } 730 William Avenue } Winnipeg, Manitoba, Canada, R3E 0W3 } e-mail: paul_hazelton-at-umanitoba.ca } Phone:204-789-3313 } Pager:204-931-954 } Cell:204-781-1502 } Fax:204-789-392 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 13:22:55 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rpowell-at-nanoprobes.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 12, 2004 at 12:22:51 ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Microscopy] Anti-GFP and anti-biotin Abs for TEM (vendor reply)
Question: Hello Michael:
[Vendor reply...] We (Nanoprobes, Inc,) make an anti-biotin-IgG gold conjugate using our 1.4 nm Nanogold particle; we also offer streptavidin-Nanogold. You don't say why the streptavidin-gold didn't work - if you could let us have some details, we may have some suggestions.
In a discussion I had with a service rep for EDAX Inc., the calibration is nothing more than setting the A/D [Analog to digital] converter's scale and offset. It doesn't change anything with respect to the detector itself.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Wednesday, May 12, 2004 11:57 AM To: Microscopy-at-msa.microscopy.com
Just guessing, but I would suppose the energy calibration is a rather simplistic algorithm. I am pretty sure that is much easier to work with a symmetric K peak and pick off the centroid than it is to deconvolute the L peaks to find the energy of the alpha line.
Presumably the effect of the background slope and curvature is negligible compared to the peak intensity if all they have to do is get a peak energy.
Warren
At 04:57 AM 5/12/2004, "michael shaffer" wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 13:26:36 2004
I've had great results on cryo-sections with a rabbit anti-GFP antibody from Molecular Probes: cat# 11122 Works well at dilutions ranging from 1:50-1:200. Low background. Hope it works for you. Best
Marc
On Wednesday, May 12, 2004, at 10:39 AM, Michal Jarnik wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } I am trying to label the GFP in yeast on either Lowicryl sections or } Tokuyashu cryosections. So far, we have had rather poor labeling/high } background. I would appreciate any recommendations on a particular } commercially available antibody and the fixation protocol. } } Also, we would need to label a biotin-tagged protein. We were trying } Streptavidin - gold, but to little success. Is there a good } anti-biotin antibody (or, even better, anti-biotin/gold conjugate) } people have used for this purpose before? } Any recommendations would be highly appreciated. } Michael Jarnik, } EM Facility } Fox Chase Cancer Center } Philadelphia, PA } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 13:32:23 2004
The Microscopy Lab in our Research Center has 3 SEM's, 1 TEM and 1 EMPA (with a second due in November) in addition to 4 XRD instruments. Our main function has been as a problem solving facility to our researchers as well as our field service engineers. Data produced by our lab is usually issued as reports with images, spectra and graphs as attachments. Management has directed us to implement a LIMS by the end of the year. Their requirements include the ability of the researchers and field service engineers to log samples in from their locations and the tracking of the sample from the arrival in our lab to the issuance of data. It must also be able to archive the data as well. For us this means literally hundreds of images, spectra and reports each month that must be categorized and saved. The researchers and engineers must also be able to access the data from their location. This will probably mean a web based system since our facilities cover the globe.
Has anyone out there been faced with a similar request? Our main hurdle is convincing management that ours is not a "sample in - number out" type of lab, and that most LIMS packages out there are designed for that type of simple operation. If you have implemented such a system, was it an off the shelf package, a custom designed package or a combination? If you want to respond to me directly that would be fine. I am not interested in "system bashing" just some helpful suggestions on how to make management happy without losing my sanity.
we like the bethyl labs mouse anti-biotin then follow that with goat anti-mouse conjugated to gold. it is much better than streptavidin gold. i am looking for a gfp ab so let me know what works.
At 10:39 AM 5/12/2004 -0400, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
The Moran Scientific EDS software can use the Cu L and K peaks (or any others the user chooses) for the peak position calibration. I do this.
It would be a bit more correct not to use L peaks, because, as Shaf said, it's an unresolved group of lines, so the resolution that gets calculated at the low energy end isn't quite correct, but unless you're using virtual standards, that doesn't matter.
But hey, even the Al K peak is the Ka1 plus the Ka2 plus the Kb anyway, on an EDS.
cheers
rtch
Date sent: Wed, 12 May 2004 08:48:26 -0400 } From: qualityimages {qualityimages-at-netrax.net} To: michael shaffer {michael-at-shaffer.net} , Microscopy {Microscopy-at-Sparc5.Microscopy.Com}
Dear Michael, I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La because those two elements are easy to find in most EM labs and the old Be-window detectors did not always see Cu La well. Of course, the accuracy of your calibration will depend on the separation of your peaks and their accurate location. Ka peaks are sharper than La peaks, so it may improve your calibration accuracy to use two Ka peaks. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 12, 2004 2:57 AM
Michael,
In response to your question, most, if not all, EDX manufacturers today including RÖNTEC find it necessary to use only one specified energy line plus the zero peak. This is because linearity is so high, that a 3-point calibration (two specified spectral lines + zero peak) for almost all cases is not beneficial.
Martin Rohde RÖNTEC GmbH RÖNTEC USA, Inc. www.rontec.com
} On May 12, 2004, at 2:57 AM, michael shaffer wrote:
} Out of curiosty, why would a calibration use these } 2 peaks rather than CuK and CuL?? Understandably, } the L-line would be a convolution of the L-family, } but I would think the Al K line is in a more difficult } location relative to the slope in the continuum(???) } Is there any preference with respect to EDX window } type? (e.g., Be v UTW) } } } I've thought that for a long time but have yet to find a } software-based calibration that can use the 2 Cu peaks. Maybe the } vendors can speak to this.
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 17:04:32 2004
It's me again. Actually I have used an anti-biotin that worked well for immuno-EM. It's from Rockland, cat # 200-4198. It's a rabbit antibody, IgG fraction.
I too had problems using streptavidin-gold. The reason for this, I think, is a lack of stability of the conjugate. Don't forget that when you purchase a gold conjugate, it might have sat for months on a refrigerator shelve and the proteins have either been degraded or fallen off the gold particles!
In my experience, good sources for antibodies that work at the EM level are Cappel (bought a a few years back by ICN - now called MP Biomedicals), Rockland (to a lesser extent), and Molecular Probes. I only use protein A gold for my labelings, and I certainly avoid gold conjugates from Ted Pella that have not worked for me even once in the last 5 years! Good luck
Marc
On Wednesday, May 12, 2004, at 01:59 PM, Michal Jarnik wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Sorry, I would try to make it clearer. We need to localize a viral } protein which has been biotin-tagged and expressed in cell culture. I } expected it should be no problem to label it with Streptavidin } conjugate, but it is not that easy, it seems (even according to some } other people). So, as we do not have a good antibody to the viral } protein, I am considering an indirect labeling with anti-biotin } antibody and a secondary conjugate. There are tons of commercially } available anti-biotin antibodies, but certainly not all of them will } work with EM protocols - so I was interested whether somebody has } experience with some of these. } } Thanks, } } Michael } } paul r hazelton wrote: } } } michael } } } } i', sorry, but your second question is not clear to me. are you } } simply } } trying to label your protein with gold, or is this a detection step } } where you have already used the antibody to react with its antigen and } } now want to see if where the antigen is? also, do you have any } } antibody } } which is not tagged with biotin and that you could use for tagging } } with } } either nanogold or directly onto larger gold labels? finally, could } } you } } do an indirect reaction using gold tagged antibody to the host species } } antibody you are using, eg, say the biotin-antibody is an IgG species, } } raised in mouse, could you come back with colloidal gold:anti mouse } } IgG? } } } } paul } } } } Paul R. Hazelton, PhD } } Electron Microscope Unit } } University of Manitoba } } Department of Medical Microbiology } } 531 Basic Medical Sciences Building } } 730 William Avenue } } Winnipeg, Manitoba, Canada, R3E 0W3 } } e-mail: paul_hazelton-at-umanitoba.ca } } Phone:204-789-3313 } } Pager:204-931-954 } } Cell:204-781-1502 } } Fax:204-789-392 } } } } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 17:44:13 2004
I believe the real object of this is to find two peaks separated far enough in energy to allow the A/D converter to calibrate over a broad range. The broader the less the error. From an electronic point of view, it doesn't matter which peaks are used, just that they have significant separation. The electronics doesn't know the difference. It goes from peak height to a position in energy which corresponds to a voltage which gates a clock. I'm babbling, only electrical engineers worry about these things.
Any EDX vendors care to comment?
Peter
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Wednesday, May 12, 2004 4:31 PM To: michael shaffer Cc: Microscopy
Dear Michael, I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La because those two elements are easy to find in most EM labs and the old Be-window detectors did not always see Cu La well. Of course, the accuracy of your calibration will depend on the separation of your peaks and their accurate location. Ka peaks are sharper than La peaks, so it may improve your calibration accuracy to use two Ka peaks. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 12, 2004 2:57 AM
Dear Peter, Your age is showing, the electronics are all purely digital, now. No A/D converter needed. The only reason to use two Ka peaks instead of a Ka and an La peak are that the Ka peaks are sharper and so the peak centroid is more precisely located. The software calibration sets the zero point and the amplifier gain to a fraction of a channel. With the newer, higher-resolution detectors, more quant precision and lower detection limit is possible, but the calibration is more critical, since a tiny shift of the peak throws off all the quant calculations. I find my self calibrating every month where I used to do it once a year with the old A/D system, but my results are much better. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "Tomic, Peter (Peter)" {ptomic-at-agere.com} To: "Mary Mager" {mager-at-interchange.ubc.ca} ; "michael shaffer" {michael-at-shaffer.net} Cc: "Microscopy" {microscopy-at-MSA.microscopy.com} Sent: Wednesday, May 12, 2004 3:49 PM
Dear Michael, I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La because those two elements are easy to find in most EM labs and the old Be-window detectors did not always see Cu La well. Of course, the accuracy of your calibration will depend on the separation of your peaks and their accurate location. Ka peaks are sharper than La peaks, so it may improve your calibration accuracy to use two Ka peaks. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 12, 2004 2:57 AM
} I find my self calibrating every month where I used to do it once a } year with the old A/D system, but my results are much better.
Once a month!
Once a year!
I do it once or twice a day.
But that's Virgoans for you. We worry a lot.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 20:07:12 2004
Hi Bettina, I have the user's manual for the Hitachi H-600. I will email you the diagrams. I remember my boss trying to fix this problem and it almost drove him crazy.
Good luck!
John Brealey Medical Scientist Electron Microscopy Unit The Queen Elizabeth Hospital IMVS - TQEH Pathology Woodville, 5011 South Australia (08) 82226612
Our physics group got a Hitachi H-600 a few years ago from biology group. We repaired it and there was no problem. But now the wire from the linkage of the specimen chamber has broken. It jumped out from the guide rolls and now we cannot move the specimen in one direction. "Nissei Sanyo Europe",the Hitachi Service Center here, demands 3500Ä for repair of this silly wire. I hope anyone can help me!!! Is there a plan (blueprint?) how to thread the wire through the rolls, so that we could repair it ourself, not perfect but in this way, that moving of the specimen would be possible?
Bettina Friedel
____________________________
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 22:08:20 2004
That is nice, but I would like more justification about why only one point is adequate. I do not buy into this hypothesis.
EDAX uses Al and Cu peaks. I think that this works very well. And, it uncovers subtle problems that otherwise would go un-noticed. I cannot see how a one point calibration is a calibration at all. Please explain this to me.
The two disparate Al-Cu peaks makes a lot of sense for calibrating an EDS. In addition to this, I do C, O, F and Al measurements with one stub. This is a separate "standard." When the results shift, if they do, I am concerned. Lately, they have shifted. And I am concerned. EDAX is on the job. Cool.
The point is to establish a set of standards and use them frequently. When the results change, contact the manufacturer. They will not know your situation unless you tell them.
gary g.
At 02:52 PM 5/12/2004, you wrote: } Michael, } } In response to your question, most, if not all, EDX manufacturers today } including RÖNTEC find it necessary to use only one specified energy line } plus the zero peak. This is because linearity is so high, that a 3-point } calibration (two specified spectral lines + zero peak) for almost all cases } is not beneficial. } } Martin Rohde } RÖNTEC GmbH } RÖNTEC USA, Inc. } www.rontec.com } } } On May 12, 2004, at 2:57 AM, michael shaffer wrote: } } } Out of curiosty, why would a calibration use these } } 2 peaks rather than CuK and CuL?? Understandably, } } the L-line would be a convolution of the L-family, } } but I would think the Al K line is in a more difficult } } location relative to the slope in the continuum(???) } } Is there any preference with respect to EDX window } } type? (e.g., Be v UTW) } } } } } I've thought that for a long time but have yet to find a } } software-based calibration that can use the 2 Cu peaks. Maybe the } } vendors can speak to this.
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 02:22:32 2004
The topic shifted a bit but with interesting points !
About that calibration questions, my few (Euro) cents :
No ADC ? Of coarse, an ADC is still necessary, but just after the (still analogic) preamp, and no more after the hole ( former analogic) counting unit.
Two points or more for energy calibration ? First point, the zero. I suppose that all today sold systems, like that we have, have a dynamic zero measurement, done on the noise peak. So nothing to do with the zero, it is continously corrected. I suppose too that the energie range of the (numerical) MCA is linear. So one (measured) peak in the midlle of the range, that is between 6 and 12 keV will be good, as said a k line will be sharper. Fe, Co or Cu... On our first Tracor NS 880, it was the first job every morning, to play with the the gain and offset trimpot, on two lines. With 150 eV resolution and a Be window, Al K was better tha Cu L (+ Cu K for high energy). A good way to put your nerves in condition to welcome your boss with a smile, when he came with a stupid question !
How often is it necessary to calibrate it ? Depends of the stability of the system and of the accuracy wanted. But realy, the need of a new calibration will be easily seen when one look the currently acquired spectra. Particulary when one work with the same family of coumpound. A shift in energy, whith a good statistic, if not been quantified, can be well detected with the eyes ! It's too with the eyes on the spectrum, that one can apreciated the real presence of a trace element, even when the soft say that you have 0.000005 % of element Xy (without overlapping, of coarse) !
And the embeding of my standards ? Nothing more to suggest ? My tests with the epoxy I have show that it is not fluid enough. So, much bulbes...
Jacques
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Resisted the urge to jump in before now, but can't any longer.
Your reasons for use of two K peaks is good, might I also add that K peaks will also offer better statistics as their count rates will be higher, thus distinguishing them better from noise and background. Most software these days allows the use of any two peaks. The choice of calibration peaks should be made on the materials you normally analyze. There can be non-linearities in the detector, pre-amplifier and amplifier responses that could be a problem if you choose peaks that are too widely separated. Aluminum and copper are commonly used because they fall within the range of elements commonly analyzed. If, on the other hand, you commonly are analyzing lanthanide's or actinides, you might want to choose peaks of higher atomic number.
Problem is that we still have to render the avalanche pulse output from a silicon detector, that merges millions of electron transfers into a single pulse who's voltage peak represents energy of the x-ray photon causing it, to a digital format. That pretty much is the definition of an analog to digital converter. This function is not abandoned in current designs, only performed earlier in the electronics. There is still an A/D converter in the signal chain, it is just faster and controlled by a DSP.
Modern designs, because of increased time resolution of processors and A/D converters, use digital signal processors (DSP) to provide many of the functions that were previously done with dedicated electronics. This trade-off has its own set of problems. Firmware is now responsible for the identification of peaks, dead time and pulse pile-up and there may be cases where that firmware could use a little tuning. This trend has resulted in manufacturer's claims of greater counting rates and resolution but also results in greater uncertainties as manufacturers strive to meet those claims. If you are finding that you have to calibrate your EDS system more frequently that is probably because of those uncertainties.
The recent trend has been to blame the inadequacies of EDS analyses on the interpretation of the signals from the detectors rather than those signals themself. In truth, we seem to have, for the moment, stretched the limit of detector technology and perhaps over-stretched the detection of those signals. The need for more frequent calibration may well be because the state of the art in pulse measurement has exceeded the state of the art in detector technology.
This thread has bifurcated into one that addresses the basic energy calibration of the detector and electronics and one that addresses the similar matrix calibrations required for good quantitative analysis. In regards to that, let me say that these are two very different, yet interrelated, areas. These are some of the most accurate measurements made in common laboratory settings and deserve some common understanding. In one case, we are calibrating our instrumentation to simplistic physical properties we understand. In the other, we are accommodating our instruments to complex interactions that we can't completely explain.
EDS calibration involves finding two reliable points that we can use to establish a linear correction to the corresponding digital values that are delivered to the computer doing the analysis. While the underlying assumption is that this is a linear relationship, that's not necessarily so. That assumption is generally made in the software that then uses this data to produce results that claim to be quantitative using either standards based or standardless algorithms.
Similar matrix calibration involves the use of well characterized standards that are of a range of composition similar to that expected in the unknown sample to provide a refinement of the simplistic linear corrections mentioned above. These standards and corrections allow us to improve our quantitative estimations by narrowing the range of linear corrections used. In reality, the interelement interactions in any particular sample can not be completely derived from the actions of their individual components at the present time.
This is still an empirical science, like it or not. As such, we have to continually accommodate our instrumentation to the task rather than to an ideal. Those accommodations will continue to be a problem and a limitation to the results we derive.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Wednesday, May 12, 2004 6:09 PM, Mary Mager [SMTP:mager-at-interchange.ubc.ca] wrote: } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Dear Peter, } Your age is showing, the electronics are all purely digital, now. No A/D } converter needed. The only reason to use two Ka peaks instead of a Ka and an La } peak are that the Ka peaks are sharper and so the peak centroid is more } precisely located. The software calibration sets the zero point and the } amplifier gain to a fraction of a channel. With the newer, higher-resolution } detectors, more quant precision and lower detection limit is possible, but the } calibration is more critical, since a tiny shift of the peak throws off all the } quant calculations. I find my self calibrating every month where I used to do it } once a year with the old A/D system, but my results are much better. } Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. } Regards, } Mary Mager } Electron Microscopist } Department of Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchange.ubc.ca } } ----- Original Message ----- } } From: "Tomic, Peter (Peter)" {ptomic-at-agere.com} } To: "Mary Mager" {mager-at-interchange.ubc.ca} ; "michael shaffer" } {michael-at-shaffer.net} } Cc: "Microscopy" {microscopy-at-MSA.microscopy.com} } Sent: Wednesday, May 12, 2004 3:49 PM } Subject: [Microscopy] RE: Re: RE: Re: EDS standards embedding } } } I believe the real object of this is to find two peaks separated far enough in } energy to allow the A/D converter to calibrate over a broad range. The broader } the less the error. From an electronic point of view, it doesn't matter which } peaks are used, just that they have significant separation. The electronics } doesn't know the difference. It goes from peak height to a position in energy } which corresponds to a voltage which gates a clock. I'm babbling, only } electrical engineers worry about these things. } } Any EDX vendors care to comment? } } Peter } } -----Original Message----- } } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] } Sent: Wednesday, May 12, 2004 4:31 PM } To: michael shaffer } Cc: Microscopy } Subject: [Microscopy] Re: RE: Re: EDS standards embedding } } } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Dear Michael, } I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka } and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La } because those two elements are easy to find in most EM labs and the old } Be-window detectors did not always see Cu La well. Of course, the accuracy of } your calibration will depend on the separation of your peaks and their accurate } location. Ka peaks are sharper than La peaks, so it may improve your calibration } accuracy to use two Ka peaks. } Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. } Regards, } Mary Mager } Electron Microscopist } Department of Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchange.ubc.ca } } ----- Original Message ----- } } From: "michael shaffer" {michael-at-shaffer.net} } To: "Gary Gaugler" {gary-at-gaugler.com} } Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} } Sent: Wednesday, May 12, 2004 2:57 AM } Subject: [Microscopy] RE: Re: EDS standards embedding } } } } } } } } ------------------------------------------------------------------------ ------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------ ------ } - } } } } Gary writes ... } } } } } The "normal" calibration of EDS is done at Al } } } K alpha (1.486KeV) and Cu K alpha (8.040KeV). } } } ... } } } } Out of curiosty, why would a calibration use these } } 2 peaks rather than CuK and CuL?? Understandably, } } the L-line would be a convolution of the L-family, } } but I would think the Al K line is in a more difficult } } location relative to the slope in the continuum(???) } } Is there any preference with respect to EDX window } } type? (e.g., Be v UTW) } } } } cheerios ... shAf :o) } } Avalon Peninsula, Newfoundland } } www.micro-investigations.com (in progress) } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 02:47:11 2004
Most of us at some moment have to deal with the capture and storage of image sequences, either as part of a video-experiment or when doing a video time-lapse. Until 3 years ago, I did my video-microscopy on a Silicon Graphics based system. The nice features of SGI were that we could do multiple videostreams in parallel, which allowed us to do multiposition time-lapse experiments.
However, three years ago I changed to another Unix-platform (Linux) and to my unpleasant surprise, it was not so trivial anymore to keep multiple video-streams in the air during an experiment and there were some other caveats too.
In the end I decided that the only thing I needed was a simple way to concatenate individual frames into a filestream, as for me the most important issue is to put the frames where I can find them afterwards for analysis, not so much for looking a them over anetwork at high speed. The format should be simple, no overhead and frame-oriented, so even mixing frames of different types should be possible. Another thing I wanted to have, was that it should be possble to store 3D-frame movie-sequences. Lossy compression schemes are not so beneficial for image anlsysis aftwerwards, as they manipulate the frame content which causes artefacts in the analysis.
The drawback of this frame-oriented approach is that some lossy compression techniques, which require multiple frame comparison, are not possible.
I would like to know if there are other people, doing video microscopy who are also interested in this approach, maybe we could create a simple software-library (ANSI C) which we can link with software we use, so we can have a video-system designed and optimized for video-microscopy (analysis oriented instead of visualization-oriented) ?
Regards,
Peter Van Osta
Director Imaging MAIA SCIENTIFIC (formerly Union Biometrica NV) Cipalstraat 3 B-2440 Geel Belgium Tel.: +32-(0)14 570 620 Fax.: +32-(0)14 570 621 Email: pvosta-at-maia-scientific.com Website: www.maia-scientific.com A Harvard Bioscience Company
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 03:08:48 2004
Sharon, it is possible to spend a lot of money on these systems. However, I have yet to find one which offers significantly more than can be done with MSExcel spreadsheets.
MSExcel spreadsheets on a server can be used in a 'shared' format, where many people can open the same file without that irritating 'This file is being used by...' error. In the system we have here, the files can only be written to by a member of the analysis group, but read by anyone. People have to come here to give us a sample anyway, so this is not a problem. Each sample gets a line in the spreadsheet with the usual fields (date, name, analysis needed etc.). When someone is working on a job, they put their initials in the box for that analysis type; when the job is done, a date goes in the box instead. The spreadsheet is updated every time someone saves it. Each column has an 'Autofilter' at the top which means they can be searched for any of the fields. This 'master' spreadsheet contains hyperlinks to the electronic log books for each microscope or technique (also shared MSExcel spreadsheets). These are updated as the analysis happens, and here each image has a line giving sample, analysis details, operator, customer, etc.. Each line in these 'technique' spreadsheets has a hyperlink to the data file. Again, each column has an 'Autofilter' at the top which means they can be searched for any of the fields.
As a user, it is easy to operate (that 'fill down' feature of Excel allows you to copy the provious line and change one box without having to make 10 entries for each image). The only discipline needed is to keep saving the files frequently (autosaving doesn't work, if two computers try to save at the same time they can lock up). Our customers seem to be happy too - they know the status of their job and can find the data without having to hassle us. We rely on the standard software on each user's PC to be able to open the images.
I'm not sure how this could be implemented on a web-based system, but for a server it works very well. We keep track of about 4000 samples a year - and all the data files they generate - with this system, and it usually only takes a few minutes to find old data even from a vague description. I have yet another spreadsheet which allows me to count how many jobs have been done, are left to do, etc., for each analysis type. The nice thing about spreadsheets is that they are totally 'customizable', so you only keep track of what you think is important.
I'd be happy to send you an example of the files if you like. As a 'free' solution it is hard to beat!
Cheers
Richard
_______________________________ Richard Beanland Analytical Services Bookham Technology plc Caswell, Towcester, Northamptonshire NN12 8EQ UK Tel: +44 (0) 1327 356362 Fax: +44 (0) 1327 356775 http://www.bookham.com All incoming mail is filtered for spam. If you do not receive a reply from me, do not assume I have received your mail.
-----Original Message----- } From: Sharon_Goresh-at-engelhard.com [mailto:Sharon_Goresh-at-engelhard.com] Sent: 12 May 2004 19:20 To: Microscopy-at-MSA.Microscopy.Com
Fellow Microscopists,
The Microscopy Lab in our Research Center has 3 SEM's, 1 TEM and 1 EMPA (with a second due in November) in addition to 4 XRD instruments. Our main function has been as a problem solving facility to our researchers as well as our field service engineers. Data produced by our lab is usually issued as reports with images, spectra and graphs as attachments. Management has directed us to implement a LIMS by the end of the year. Their requirements include the ability of the researchers and field service engineers to log samples in from their locations and the tracking of the sample from the arrival in our lab to the issuance of data. It must also be able to archive the data as well. For us this means literally hundreds of images, spectra and reports each month that must be categorized and saved. The researchers and engineers must also be able to access the data from their location. This will probably mean a web based system since our facilities cover the globe.
Has anyone out there been faced with a similar request? Our main hurdle is convincing management that ours is not a "sample in - number out" type of lab, and that most LIMS packages out there are designed for that type of simple operation. If you have implemented such a system, was it an off the shelf package, a custom designed package or a combination? If you want to respond to me directly that would be fine. I am not interested in "system bashing" just some helpful suggestions on how to make management happy without losing my sanity.
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From MicroscopyL-request-at-ns.microscopy.com Thu May 13 05:16:19 2004
A two-step detection method is indeed a good option to visualize your biotinylated viral protein. Compared to the 2-step method the advantage of a 1-step immunodetection method, as you initially suggested, will be improved resolution. Your labeling density decreases however. As you know labeling density is inversely related to the gold particle size.
Improved resolution combined with high sensitivity can be obtained when using gold conjugates prepared with Ultra-Small gold particles. For your application we would recommend Goat anti Biotin Ultra-Small. See e.g. Müller-Höcker et al., (1998), Histochem. and Cell Biol. 109(2), 119-125.
When the protein of interest is located on the outside of the virus, in other words the biotin is accessible for the antibodies, another option may be the use of a pre-embedding labeling procedure. The use of Ultra-Small gold conjugates is a prerequisite to successfully combine appropriate labeling densities with a decent morphology. See e.g. Decossas et al, (2003) J. of Comp. Neurology 462(3), 302-314 for details on this technique.
Hope this is of help Kind regards, Peter
------------------------------------------ Michael Jarnik wrote:
Sorry, I would try to make it clearer. We need to localize a viral protein which has been biotin-tagged and expressed in cell culture. I expected it should be no problem to label it with Streptavidin conjugate, but it is not that easy, it seems (even according to some other people). So, as we do not have a good antibody to the viral protein, I am considering an indirect labeling with anti-biotin antibody and a secondary conjugate. There are tons of commercially available anti-biotin antibodies, but certainly not all of them will work with EM protocols - so I was interested whether somebody has experience with some of these.
Thanks,
Michael
----------- Peter van de Plas Aurion Costerweg 5 6702 AA Wageningen The Netherlands Phone: +31 317 497676 Fax: +31 317 415955
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 06:55:04 2004
A two-step detection method is indeed a good option to visualize your biotinylated viral protein. Compared to the 2-step method the advantage of a 1-step immunodetection method, as you initially suggested, will be improved resolution. Your labeling density decreases however. As you know labeling density is inversely related to the gold particle size.
Improved resolution combined with high sensitivity can be obtained when using gold conjugates prepared with Ultra-Small gold particles. For your application we would recommend Goat anti Biotin Ultra-Small. See e.g. Müller-Höcker et al., (1998), Histochem. and Cell Biol. 109(2), 119-125.
When the protein of interest is located on the outside of the virus, in other words the biotin is accessible for the antibodies, another option may be the use of a pre-embedding labeling procedure. The use of Ultra-Small gold conjugates is a prerequisite to successfully combine appropriate labeling densities with a decent morphology. See e.g. Decossas et al, (2003) J. of Comp. Neurology 462(3), 302-314 for details on this technique.
Hope this is of help Kind regards, Peter
------------------------------------------ Michael Jarnik wrote:
Sorry, I would try to make it clearer. We need to localize a viral protein which has been biotin-tagged and expressed in cell culture. I expected it should be no problem to label it with Streptavidin conjugate, but it is not that easy, it seems (even according to some other people). So, as we do not have a good antibody to the viral protein, I am considering an indirect labeling with anti-biotin antibody and a secondary conjugate. There are tons of commercially available anti-biotin antibodies, but certainly not all of them will work with EM protocols - so I was interested whether somebody has experience with some of these.
Thanks,
Michael
----------- Peter van de Plas Aurion Costerweg 5 6702 AA Wageningen The Netherlands Phone: +31 317 497676 Fax: +31 317 415955
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 09:29:14 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rosscac-at-aol.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, May 13, 2004 at 07:52:40 ---------------------------------------------------------------------------
Question: I am interested in purchasing a print processor and was wondering if anyone out there is using or has used either a Fujimoto Roller Transport print processor CP51 or a JOBO printlab 3504 print processor. I was looking for some feedback on these units on there durablility and functionalitiy or any other tidbits! You can respond back via email to me at rosscac-at-aol.com or at connie.a.cummings-at-gsk.com. Thanks in advance. Connie
A post doc is trying to detect GFP being expressed in plant leaves. She is fixing leaves in formaldehyde and doing a standard paraffin embedding. Her question is if GFP going to survive xylene treatments for paraffin infiltration. She plans to section the paraffin embedded blocks and look for GFP using fluorescence microscopy.
If anyone has an answer please reply back to me.
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 10:16:44 2004
Hi all, Lazy me recommended pre-coated SuperFrostPlus slides to Anne Dunn a post-doc working on antlions but the gut goo doesn't always survive her processing protocol (see her message below).
So the question is - What is the preferred slide coating method? Gelatin vs poly-L-lysine...or something else?
advice and recipes would be greatly appreciated. thanks, Beth
} Hi Beth, } } I am finding that when I use the superfrost plus slides for } my cryosections that often if there is material inside the } gut that it will wash away as I am fixing and dehydrating } my sections before hybridization (and I am not seeing any } bacteria in the gut area after hybridization). I have no idea } whether it is just something that is going to happen with } a fluid-feeding insect, or if it is the superfrost slides not } being sticky enough. I don't seem to be having a } problem with the tissue itself sticking. } } I was looking up ways to coat slides and found there is } the gelatin method and the poly-L-lysine method. } Unfortunately I am not finding any info that tells me if one } is better than the other. Do you have an opinion? I was } wondering if it is a case of one coating being better for } certain applications than another? } } I was going to go ahead and dissect out a gut or two, fix } and hybridize them, trying to see if I can find any } bacteria-like objects in the gut smooshate...to let me } know if the loss of the material in the gut is an issue or } not, but wanted to start thinking/plotting if I need to coat } some slides.
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I'm looking for a shield to exclude air movement around the stage of my Ultracut. I remember years ago having one for my Reichert OM-U3 but I don't believe I got one for my Ultracut a few years ago. I've checked a couple of the vendors and found nothing. I could find nothing at the Leica site either. Does anyone have a source for these or will I have to make my own?
Rick A. Harris, exDirector Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://microscopy.mcb.ucdavis.edu raharris-at-ucdavis.edu
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 11:41:29 2004
Richard is of course right -- you can spend a fortune on LIMS systems and for most users it may be overkill. On the other hand, the Excel sheets that Richard mentions may be too limited.
A third option, somewhat in the middle, could be to use software that supports or can read the images and other data from your software and keep track of those in an archive or database. That way you can search for keyfields, you deal with the same software for all applications, and you can distribute the images easily. Our software analySIS includes such a database. We also have a web frontend for this database so you can deal with the global issues you mentioned.
Please take a look at our website or contact me directly if you wolud like to get more information.
Mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Richard Beanland [mailto:richard.beanland-at-bookham.com] Sent: Thursday, May 13, 2004 02:14 To: 'Sharon_Goresh-at-engelhard.com' Cc: 'Microscopy-at-MSA.Microscopy.Com'
Sharon, it is possible to spend a lot of money on these systems. However, I have yet to find one which offers significantly more than can be done with MSExcel spreadsheets.
MSExcel spreadsheets on a server can be used in a 'shared' format, where many people can open the same file without that irritating 'This file is being used by...' error. In the system we have here, the files can only be written to by a member of the analysis group, but read by anyone. People have to come here to give us a sample anyway, so this is not a problem. Each sample gets a line in the spreadsheet with the usual fields (date, name, analysis needed etc.). When someone is working on a job, they put their initials in the box for that analysis type; when the job is done, a date goes in the box instead. The spreadsheet is updated every time someone saves it. Each column has an 'Autofilter' at the top which means they can be searched for any of the fields. This 'master' spreadsheet contains hyperlinks to the electronic log books for each microscope or technique (also shared MSExcel spreadsheets). These are updated as the analysis happens, and here each image has a line giving sample, analysis details, operator, customer, etc.. Each line in these 'technique' spreadsheets has a hyperlink to the data file. Again, each column has an 'Autofilter' at the top which means they can be searched for any of the fields.
As a user, it is easy to operate (that 'fill down' feature of Excel allows you to copy the provious line and change one box without having to make 10 entries for each image). The only discipline needed is to keep saving the files frequently (autosaving doesn't work, if two computers try to save at the same time they can lock up). Our customers seem to be happy too - they know the status of their job and can find the data without having to hassle us. We rely on the standard software on each user's PC to be able to open the images.
I'm not sure how this could be implemented on a web-based system, but for a server it works very well. We keep track of about 4000 samples a year - and all the data files they generate - with this system, and it usually only takes a few minutes to find old data even from a vague description. I have yet another spreadsheet which allows me to count how many jobs have been done, are left to do, etc., for each analysis type. The nice thing about spreadsheets is that they are totally 'customizable', so you only keep track of what you think is important.
I'd be happy to send you an example of the files if you like. As a 'free' solution it is hard to beat!
Cheers
Richard
_______________________________ Richard Beanland Analytical Services Bookham Technology plc Caswell, Towcester, Northamptonshire NN12 8EQ UK Tel: +44 (0) 1327 356362 Fax: +44 (0) 1327 356775 http://www.bookham.com All incoming mail is filtered for spam. If you do not receive a reply from me, do not assume I have received your mail.
-----Original Message----- } From: Sharon_Goresh-at-engelhard.com [mailto:Sharon_Goresh-at-engelhard.com] Sent: 12 May 2004 19:20 To: Microscopy-at-MSA.Microscopy.Com
Fellow Microscopists,
The Microscopy Lab in our Research Center has 3 SEM's, 1 TEM and 1 EMPA (with a second due in November) in addition to 4 XRD instruments. Our main function has been as a problem solving facility to our researchers as well as our field service engineers. Data produced by our lab is usually issued as reports with images, spectra and graphs as attachments. Management has directed us to implement a LIMS by the end of the year. Their requirements include the ability of the researchers and field service engineers to log samples in from their locations and the tracking of the sample from the arrival in our lab to the issuance of data. It must also be able to archive the data as well. For us this means literally hundreds of images, spectra and reports each month that must be categorized and saved. The researchers and engineers must also be able to access the data from their location. This will probably mean a web based system since our facilities cover the globe.
Has anyone out there been faced with a similar request? Our main hurdle is convincing management that ours is not a "sample in - number out" type of lab, and that most LIMS packages out there are designed for that type of simple operation. If you have implemented such a system, was it an off the shelf package, a custom designed package or a combination? If you want to respond to me directly that would be fine. I am not interested in "system bashing" just some helpful suggestions on how to make management happy without losing my sanity.
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From MicroscopyL-request-at-ns.microscopy.com Thu May 13 12:27:22 2004
We use silane to coat our slides, it is the standard for the histology labs in the pathology dept.
Silane = 3-aminopropyl-triethoxy-silane
Protocol:
Silane 20 ml Actenoe 2000 ml
Dip precleaned slides in Silane solution for 2 minutes Drain and wash in distilled water 2 times Drain and dry at 60 degrees C for 1 hour Store away from light and moisture.
Our sildes last for many months (6+), we make huge batches at a time.
I hope this helps you out of your sticky situation!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
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Hi all, Lazy me recommended pre-coated SuperFrostPlus slides to Anne Dunn a post-doc working on antlions but the gut goo doesn't always survive her processing protocol (see her message below).
So the question is - What is the preferred slide coating method? Gelatin vs poly-L-lysine...or something else?
advice and recipes would be greatly appreciated. thanks, Beth
} Hi Beth, } } I am finding that when I use the superfrost plus slides for } my cryosections that often if there is material inside the } gut that it will wash away as I am fixing and dehydrating } my sections before hybridization (and I am not seeing any } bacteria in the gut area after hybridization). I have no idea } whether it is just something that is going to happen with } a fluid-feeding insect, or if it is the superfrost slides not } being sticky enough. I don't seem to be having a } problem with the tissue itself sticking. } } I was looking up ways to coat slides and found there is } the gelatin method and the poly-L-lysine method. } Unfortunately I am not finding any info that tells me if one } is better than the other. Do you have an opinion? I was } wondering if it is a case of one coating being better for } certain applications than another? } } I was going to go ahead and dissect out a gut or two, fix } and hybridize them, trying to see if I can find any } bacteria-like objects in the gut smooshate...to let me } know if the loss of the material in the gut is an issue or } not, but wanted to start thinking/plotting if I need to coat } some slides.
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I am going to buy scanner for TEM negatives and considering two models: Microtek ArtixScan 1800f and Epson Expression 1680 Pro. Any advise and comments are very welcome.
Thank you in advance,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
I am looking for a replacement glass spacer for a Polaron E5100 series II 'Cool' Sputter Coater. The external diameter etched on metal collar base is 165.1 m/m. The internal diameter of the metal collar is 5.75". The height of the glass spacer is 3 and 3/16". I have been working with EBS to locate a replacement collar but would like to know if there are any other sources I could contact.
Thank you in advance for your help
aruna
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 15:30:40 2004
Dear Sylvain, I use graphite, evaporated in a high-vacuum evaporator, to make SEM samples conductive when I want to do EDS analysis. I use gold/palladium for SEM imaging without analysis, but carbon coating to minimize any interference of the coating with the EDS analysis. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "sylvain maury" {sylvain.maury-at-thalesgroup.com} To: "Microscopy community" {microscopy-at-msa.microscopy.com} Sent: Thursday, May 13, 2004 8:22 AM
} I'm looking for a shield to exclude air movement around the stage of my } Ultracut. I remember years ago having one for my Reichert OM-U3 but I } don't believe I got one for my Ultracut a few years ago. I've checked a } couple of the vendors and found nothing. I could find nothing at the } Leica site either. Does anyone have a source for these or will I have } to make my own? } } Rick A. Harris, exDirector } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } http://microscopy.mcb.ucdavis.edu } raharris-at-ucdavis.edu
Rick -
Make a hole in the bottom of a plastic veggie bag from your supermarket & rubber-band it around the binocular objective. Ugly, but it works better than a rigid plastic box.
Caroline -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 16:09:34 2004
Short and sweet: about a year ago we bought a Microtek ArtixScan 2500f (firewire); a little different from the 1800f but I don't know how much. Anyway, we like it. Great dynamic range and reasonable software (we scan into a G4 Mac running OSX).
cheers, John
******** John S. Vetrano Sr. Research Scientist Materials Structure and Performance Group Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
} ---------- } From: Dusevich, Vladimir } Sent: Thursday, May 13, 2004 11:25 AM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I am going to buy scanner for TEM negatives and considering two models: } Microtek ArtixScan 1800f and Epson Expression 1680 Pro. Any advise and } comments are very welcome. } } Thank you in advance, } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 16:33:19 2004
Hi, I would like to have an evaluation of the Denton Desk III TSC sputter coater from those who are using or have used this instrument for preparation of samples for FESEM imaging. Please direct your response to me rather than to the list.
Many thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 18:42:36 2004
I don't use C. I use Au/Pd and Pt. C is too messy.
Depending on what you are looking at, either image- or EDS/WDS-wise, your options may vary. For EDS, I just ignore the small Au/Pd peaks. The coating is only 40-60A and is typically only seen as piled up Au with Al, Si and W. With my specimens, Pd is by itself.
I suppose that if you were looking at P or Zr specimens, Au/Pd or Pt would be a problem. Perhaps the ultimate coating is Os. Someday, I would love to try one of these coaters. It ought to be great for EBSD.
But let us know what type of work you are trying to do. That may make a big difference in coating strategies.
gary g.
At 08:22 AM 5/13/2004, you wrote:
} hi all! } } a simple question : } } Does anyone have ever used graphite for sputtering or coating a SEM } sample ? } } maybe a question from a dummy boy :-) , doesn't it? } } thanx for answers... } } Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 20:43:29 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I don't use C. I use Au/Pd and Pt. C is too messy.
Depending on what you are looking at, either image- or EDS/WDS-wise, your options may vary. For EDS, I just ignore the small Au/Pd peaks. The coating is only 40-60A and is typically only seen as piled up Au with Al, Si and W. With my specimens, Pd is by itself.
I suppose that if you were looking at P or Zr specimens, Au/Pd or Pt would be a problem. Perhaps the ultimate coating is Os. Someday, I would love to try one of these coaters. It ought to be great for EBSD.
But let us know what type of work you are trying to do. That may make a big difference in coating strategies.
gary g.
At 08:22 AM 5/13/2004, you wrote:
} hi all! } } a simple question : } } Does anyone have ever used graphite for sputtering or coating a SEM } sample ? } } maybe a question from a dummy boy :-) , doesn't it? } } thanx for answers... } } Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Fri May 14 08:16:15 2004
I have an earlier model of the Epson (the 1600 Pro) and I"ve been happy with it. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Fri May 14 09:44:02 2004
Someone recently posed the same question on ListServer. We have an Epson Perfection 3200 Photo and love it. There is no holder for the standard 3 1/4 X 4 negatives, but I just put them directly on the glass and they come out fine.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] Sent: Thursday, May 13, 2004 2:26 PM To: microscopy-at-msa.microscopy.com
I am going to buy scanner for TEM negatives and considering two models: Microtek ArtixScan 1800f and Epson Expression 1680 Pro. Any advise and comments are very welcome.
Thank you in advance,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Fri May 14 16:30:39 2004
What is the cross sectional construction like? Is the die passivated? If it is, use MIL-STD-883 for GLIT. If there is metal underneath the passivation, and the passivation has voids, GLIT will show it up big time. GLIT is glassivation layer integrity test.
gary g.
At 01:10 PM 5/14/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri May 14 17:47:07 2004
Dear Pavel: my first thought is, don't coat the device. Find an accelerating voltage/tilt angle combination that will prevent of minimize charging. Next thought, coat with something besides gold. But I've done a lot of cracked devices and have never had any problems telling the cracks from the original surface. Unless they are huge cracks, the gold won't get in them to confuse the issue. Are these surface cracks or cracks going down into the surface?
AtcSEM wrote:
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-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Sat May 15 00:47:25 2004
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From MicroscopyL-request-at-ns.microscopy.com Sun May 16 13:24:22 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zr zrzhang_xrli-at-ybb.ne.jp) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, May 15, 2004 at 21:16:24 ---------------------------------------------------------------------------
I need to prepare TEM samples from Ag and Ag-Pd(10pct) alloy sheets. May you suggest me any electrolytes for twin-jet electropolishing and the corresponding conditions? No toxic chemicals would be used.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jtrynak-at-SciGenium.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 12, 2004 at 18:22:09 ---------------------------------------------------------------------------
Email: jtrynak-at-SciGenium.com Name: JT Rynak
Organization: SciGenium
Title-Subject: [Microscopy] [Filtered] Opening - New England Regional - Imaging Sales
Question: New England Regional - Imaging Sales
5+ years experience selling imaging / visualization capital equipment into Research Market space. Ideal person will have a rolodex of PI and Lead Vivo research scientists. BS/MS is Biology or Zoology
If you have any further questions feel free to contact me at 617-661-1067 or via email at jtrynak-at-scigenium.com
Having looked at more fractures in silicon and GaAs devices than I care to remember, it's not generally a problem finding them and as often with optical means as with EM.
If you would kindly provide the list with more specifics about your samples and the nature of these "cracks?" That is, are they along crystal planes or fractures that travel at random angles with respect to the surface? Fractures, or cleaves perpendicular to the die surface, are difficult to detect with EM but apparent with visible light microscopy. How separated are both sides of the crack and why do you need to see into them? Sounds like you only need to identify whether they exist or not. How long are the cracks?
I have plenty of EM's I'd be happy to send you images of samples that have fractured for various reasons, and there are multiple root causes of this failure mechanism. I'm sure if you provide a little more detail you'll get plenty of good suggestions on how to image your samples.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Friday, May 14, 2004 4:10 PM To: MicroscopyListServer (MicroscopyListServer)
Dear microscopy experts I need your expertise.
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Sun May 16 19:26:15 2004
We are in the process of justifying, to the University, a proposal to purchase an ESEM. In prepapring the business case I am including the ten year financials for both FEG and Tungtsen filament machines. Given that we have to break even in that time, including covering our depreciation, the Tungsten instrument is easier to justify in a purely financial sense.
From a scientific and operational viewpoint, though, I was hoping some of you could provide opinions on the relative merits of Tungsten or FEG. Our current conventional SEM is a Philips XL-30 S-FEG so we already know of the costs associated with the emitter.
The range of samples we will be required to put in our ESEM will be anything you oculd possibly imagine from the Faculties of Engineering, Science and Medicine. Samples will inlcude food materials (big dairy industry in New Zealand), polymers, conducting polymers, mammalian tissue, bacteria, fruit and veg, fish and chips, superconductors, ceramic coatings and bees knees. We will definitely need a hot-stage.
If you would like to contact me off list with your opinions please feel free.
Kind regards Bryony James Research Centre for Surface and Materials Science Univetrsity of Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 07:57:52 2004
Thank you all for the responses that I have received.
Just to clarify: We had multiple field failures of the chips. The SEM examination revealed some sort of coating on the fracture surfaces, which I believe is the urethane coating. The transition from the side wall to the fracture surface, on some of the chips, suggests that the chips could have had a small cracks prior urethane coating.
Now, I am looking at the "Virgin" chips for any evidence of cracking at the side wall and diaphragm transition, but my problem is that the Gold coating does not coat the area well enough for me to resolve the crack. Due to lack of coating that area is charging and I see only a black line. I have coated it at least 3 times and rotate the chips to insure good coatings in those areas, but it was not enough.
After SEM examination I would also like to cross section the chip to check for the evidence of pre-cracks, but I am afraid to introduce the cracks myself. Could you help me with a proper preparation procedure of the cross sections? We have metallography lab. equipment and nothing special to prepare the chip cross sections.
I have uploaded some more photos of the fracture surfaces of the failed chips and Virgin chip that I am trying to image now. http://www.atclabs.com/images/temp/MicrList_files/frame.htm
Sorry, it might be a little slow at showing the images due to their size.
Thanks for your help again,
Pavel
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com]
What is the cross sectional construction like? Is the die passivated? If it is, use MIL-STD-883 for GLIT. If there is metal underneath the passivation, and the passivation has voids, GLIT will show it up big time. GLIT is glassivation layer integrity test.
gary g.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu]
Do you get charging at the crack due to a gap in the coating? Otherwise,you should be able to see the break.
-----Original Message----- } From: Jim Quinn [mailto:jquinn-at-www.matscieng.sunysb.edu] Pavel
Your image is very low mag, so it is difficult to judge anything.
I suggest that you use a sacrificial chip and coat it with about 10nm of gold. If your coater does not rotate the sample, then you should manually rotate the sample at 45 degrees atleast twice. This is will help the gold to get into the perpendicular surfaces.
Next use a Robinson BS detector. 10 or 15kV is sufficient.
Once you image the cracks, then you can go about trying to image without the gold. Variable pressure and voltage will reduce charging in uncoated samples.
JQ
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Pavel;
Having looked at more fractures in silicon and GaAs devices than I care to remember, it's not generally a problem finding them and as often with optical means as with EM.
If you would kindly provide the list with more specifics about your samples and the nature of these "cracks?" That is, are they along crystal planes or fractures that travel at random angles with respect to the surface? Fractures, or cleaves perpendicular to the die surface, are difficult to detect with EM but apparent with visible light microscopy. How separated are both sides of the crack and why do you need to see into them? Sounds like you only need to identify whether they exist or not. How long are the cracks?
I have plenty of EM's I'd be happy to send you images of samples that have fractured for various reasons, and there are multiple root causes of this failure mechanism. I'm sure if you provide a little more detail you'll get plenty of good suggestions on how to image your samples.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Friday, May 14, 2004 4:10 PM To: MicroscopyListServer (MicroscopyListServer)
Dear microscopy experts I need your expertise.
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 09:11:57 2004
Thank you so much for all of your advice. I received a large number of great suggestions and almost everyone suggested using anti GFP antibody and we are certainly going to go that route.
Thanks again and this is truly a great resource.
Best wishes,
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 11:36:59 2004
1) Can you say "stress concentration"? My academic background is in Engineering Science and Mechanics. It looks like you have some very sharp corners where the diaphragm meets the substrate. That is trouble waiting to happen. I cannot tell if there is any pre-crack there. There might be hints of one in slide 43. Either way, that geometry is going to lead to problems. The stresses will be highest at the edges of the diaphragm. I think you would prefer a gradual change in geometry there to avoid any concentration and to help reinforce the edges. I don't know the fabrication process, and it might be hard to implement, but it might be worth investigating.
Slides 38 and 42 show some jagged surfaces. I guess those are the edges of the substrate and the diaphragm is on the bottom. If so, the nose of the jaggie jutting out further into the opening will be a point of stress concentration.
You may also want to consider round openings in the substrate. The corners of the square holes don't help and might hurt. The highest stresses should be in the middle of the edges.
2) You haven't said how prematurely these chips failed. Are they subjected to a cyclic load? What fraction of their design load were they subjected to? There may be hints of fatigue failure (slide 34?), but it seems to be rather short cycle fatigue if it is fatigue at all. I have seen glass break with such fractures in overload.
3) Slides 37, 40 and 41 show dark corners. I think that is the normal appearance for an interior corner in secondary mode. An exterior corner tends to show up bright in SE mode.
4) I can't quite tell where many of the photos were taken (e.g., slides 13 and 39). I usually use a three-fold step in magnification to easily follow from one view to another. Also, the progressions 10, 30, 100, 300 or 15, 50, 150, 500 are easy to remember and to standardize on.
5) Did you copy these images into Powerpoint to make this presentation? My computer took quite a while to load each slide, and I have a fast connection. We routinely record our SEM images at 1024-pixels across and store the files as JPGs. (Keep reading for the rationale.) Those files are maybe 300 KB in size. If I use the Insert, Image, From_file function to add the image to my document, it retains the JPG compression and gray nature and requires only 300 KB. But, if I open those files in an imaging program and copy them over they expand. The computer has the image rendered as a bitmap in full color, the image gets copied with no compression and it now consumes 2400 kB (1024x768 pixels times 3-bytes-per-pixel). That 8-fold difference in size is noticeable. I know more and more users have broadband connections; however these order-of-magnitude expansions start to add up. {g}
Hope this helps.
Warren
At 08:06 AM 5/17/2004, you wrote:
} Thank you all for the responses that I have received. } } Just to clarify: } We had multiple field failures of the chips. The SEM } examination } revealed some sort of coating on the fracture surfaces, which I believe is } the urethane coating. The transition from the side wall to the fracture } surface, on some of the chips, suggests that the chips could have had a } small cracks prior urethane coating. } } Now, I am looking at the "Virgin" chips for any evidence of cracking } at the side wall and diaphragm transition, but my problem is that the Gold } coating does not coat the area well enough for me to resolve the crack. Due } to lack of coating that area is charging and I see only a black line. } I have coated it at least 3 times and rotate the chips to insure good } coatings in those areas, but it was not enough. } } After SEM examination I would also like to cross section the chip to check } for the evidence of pre-cracks, but I am afraid to introduce the cracks } myself. Could you help me with a proper preparation procedure of the cross } sections? We have metallography lab. equipment and nothing special to } prepare the chip cross sections. } } I have uploaded some more photos of the fracture surfaces of the failed } chips and Virgin chip that I am trying to image now. } http://www.atclabs.com/images/temp/MicrList_files/frame.htm } } Sorry, it might be a little slow at showing the images due to their size. } } Thanks for your help again, } } Pavel
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
We look mostly at hydrated biological samples. We have a tungsten gun XL30 ESEM TMP. For looking at bacteria and cells a FEGESEM would be superior in terms of reduced damage and improved resolution. I find it hard to get medium/high resolution with wet biological samples. I feel FEGESEM would make this a lot easier.
Dave
On Mon, 17 May 2004 17:21:33 +1200 Bryony James {b.james-at-auckland.ac.nz} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello listers, } } We are in the process of justifying, to the University, a proposal to } purchase an ESEM. In prepapring the business case I am including the } ten year financials for both FEG and Tungtsen filament machines. Given } that we have to break even in that time, including covering our } depreciation, the Tungsten instrument is easier to justify in a purely } financial sense. } } From a scientific and operational viewpoint, though, I was hoping some } of you could provide opinions on the relative merits of Tungsten or FEG. } Our current conventional SEM is a Philips XL-30 S-FEG so we already know } of the costs associated with the emitter. } } The range of samples we will be required to put in our ESEM will be } anything you oculd possibly imagine from the Faculties of Engineering, } Science and Medicine. Samples will inlcude food materials (big dairy } industry in New Zealand), polymers, conducting polymers, mammalian } tissue, bacteria, fruit and veg, fish and chips, superconductors, } ceramic coatings and bees knees. We will definitely need a hot-stage. } } If you would like to contact me off list with your opinions please feel } free. } } Kind regards } Bryony James } Research Centre for Surface and Materials Science } Univetrsity of Auckland } New Zealand } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 13:37:31 2004
We are an independent commercial lab serving general industry. Unless directed by a court order or we suspect misuse of our findings, all information is strictly confidential to our client.
We have a situation with a small project we just completed. We detected the presence of a toxic compound in product. While not the intended use, it is conceivable that this product could be put into one's mouth which would present a health concern.
Upon discovery of this compound, I called our client immediately and found out that they submitted competitors' products. Our client is certainly aware of this now, but they are not in a position to disclose to their competitors that they were analyzing their products. Given that these products are not intended to have oral contact, I doubt that the FDA has any applicable regulations regarding the composition.
Any thoughts?
Alan Stone ASTON Metallurgical Services
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 16:10:25 2004
Dear Bryony, The combined FE and ESED instruments are very pricey, expensive to maintain and, since you already have a FE, perhaps a bit redundant. We have a two-year-old tungsten variable-pressure and it is a popular workhorse that is in constant use. It can do high resolution or ESED but not both at the same time. If you need both at the same time, then you need FE-ESED. BTW, the thing we would most like to have is a cold stage, to look at hydrated samples without having to go to higher pressures. Good luck. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Bryony James" {b.james-at-auckland.ac.nz} To: {Microscopy-at-msa.microscopy.com} Sent: Sunday, May 16, 2004 10:21 PM
Alan;
It sounds like you need legal advice and not scientific advice. You should consult with your corporate counsel on this issue.
Competitor analysis is done all the time and I don't see any issues with that. What you do with the data afterwards may be another matter and open to litigation so see the attorney.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: Alan Stone [mailto:as-at-astonmet.com] Sent: Monday, May 17, 2004 2:47 PM To: Microscopy-at-MSA.Microscopy.Com
We are an independent commercial lab serving general industry. Unless directed by a court order or we suspect misuse of our findings, all information is strictly confidential to our client.
We have a situation with a small project we just completed. We detected the presence of a toxic compound in product. While not the intended use, it is conceivable that this product could be put into one's mouth which would present a health concern.
Upon discovery of this compound, I called our client immediately and found out that they submitted competitors' products. Our client is certainly aware of this now, but they are not in a position to disclose to their competitors that they were analyzing their products. Given that these products are not intended to have oral contact, I doubt that the FDA has any applicable regulations regarding the composition.
Any thoughts?
Alan Stone ASTON Metallurgical Services
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 07:02:20 2004
If you really need a x-section without the issues with mechanical polishing, try using an FIB [Focused Ion Beam].
Peter
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Monday, May 17, 2004 9:07 AM To: MicroscopyListServer (MicroscopyListServer)
Thank you all for the responses that I have received.
Just to clarify: We had multiple field failures of the chips. The SEM examination revealed some sort of coating on the fracture surfaces, which I believe is the urethane coating. The transition from the side wall to the fracture surface, on some of the chips, suggests that the chips could have had a small cracks prior urethane coating.
Now, I am looking at the "Virgin" chips for any evidence of cracking at the side wall and diaphragm transition, but my problem is that the Gold coating does not coat the area well enough for me to resolve the crack. Due to lack of coating that area is charging and I see only a black line. I have coated it at least 3 times and rotate the chips to insure good coatings in those areas, but it was not enough.
After SEM examination I would also like to cross section the chip to check for the evidence of pre-cracks, but I am afraid to introduce the cracks myself. Could you help me with a proper preparation procedure of the cross sections? We have metallography lab. equipment and nothing special to prepare the chip cross sections.
I have uploaded some more photos of the fracture surfaces of the failed chips and Virgin chip that I am trying to image now. http://www.atclabs.com/images/temp/MicrList_files/frame.htm
Sorry, it might be a little slow at showing the images due to their size.
Thanks for your help again,
Pavel
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com]
What is the cross sectional construction like? Is the die passivated? If it is, use MIL-STD-883 for GLIT. If there is metal underneath the passivation, and the passivation has voids, GLIT will show it up big time. GLIT is glassivation layer integrity test.
gary g.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu]
Do you get charging at the crack due to a gap in the coating? Otherwise,you should be able to see the break.
-----Original Message----- } From: Jim Quinn [mailto:jquinn-at-www.matscieng.sunysb.edu] Pavel
Your image is very low mag, so it is difficult to judge anything.
I suggest that you use a sacrificial chip and coat it with about 10nm of gold. If your coater does not rotate the sample, then you should manually rotate the sample at 45 degrees atleast twice. This is will help the gold to get into the perpendicular surfaces.
Next use a Robinson BS detector. 10 or 15kV is sufficient.
Once you image the cracks, then you can go about trying to image without the gold. Variable pressure and voltage will reduce charging in uncoated samples.
JQ
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Pavel;
Having looked at more fractures in silicon and GaAs devices than I care to remember, it's not generally a problem finding them and as often with optical means as with EM.
If you would kindly provide the list with more specifics about your samples and the nature of these "cracks?" That is, are they along crystal planes or fractures that travel at random angles with respect to the surface? Fractures, or cleaves perpendicular to the die surface, are difficult to detect with EM but apparent with visible light microscopy. How separated are both sides of the crack and why do you need to see into them? Sounds like you only need to identify whether they exist or not. How long are the cracks?
I have plenty of EM's I'd be happy to send you images of samples that have fractured for various reasons, and there are multiple root causes of this failure mechanism. I'm sure if you provide a little more detail you'll get plenty of good suggestions on how to image your samples.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Friday, May 14, 2004 4:10 PM To: MicroscopyListServer (MicroscopyListServer)
Dear microscopy experts I need your expertise.
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 07:15:34 2004
Hello, We want to image, by SEM, the surface of a HfO2 layer deposited on Si. Is there a method to reveal the structure of the layer ? for instance chemical etching or others.... We have tried on the as grown layer but we can see nothing, the image is very smooth. By means of X-ray diffraction we know that we have very small crystals, just a few nanometers. Now we wonder wether there are larger structures than the crystals. We expect something like polycrystalline grains... Thank you for your help Regards
-- Claude GRATTEPAIN / Bernard SERVET
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From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:17:11 2004
We would like to get a centrifuge to separate viruses. What are you using in your TEM laboratory? Suggestions and hints will be helpful. Thanks
Since my UB mail address is not reliable please send a copy of all urgent mail to
coetzeesh-at-yahoo.co.uk
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana Phone : +267 355 2462/5222 Mobile : +267 714 55701 (Now active) Fax : +267 318 5097 e-mail : coetzees-at-mopipi.ub.bw
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:46:08 2004
I wanted to see if anyone has had any experience scanning negatives with the Canon Canoscan 9900F. I have a low end Canoscan that I use for documents and the occasional photograph, which has given me very good results, but is not set up for negatives. I have seen the recent posts about the Epson scanner, but I am trying to keep it under a $1000 so that I do not have to treat it as a capital purchase. I have tried for the last two years to get a Coolscan 8000 on the capital budget with no luck, and do not see any scanner above $1000 as an option right now.
Thanks, Steve
Steven Lee Chief Technologists Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 750246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu
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From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:47:42 2004
In my experience medium/high magnifications for wet specimens are limited mostly by specimens themselves and not by a type of emitter. While I can get a nice picture of gold on carbon at x100k, for most of my "real life" specimens magnification is limited by x20k (or less).
Vladimir
} We look mostly at hydrated biological samples. We have a } tungsten gun XL30 ESEM TMP. For looking at bacteria and } cells a FEGESEM would be superior in terms of reduced } damage and improved resolution. I find it hard to get } medium/high resolution with wet biological samples. I feel } FEGESEM would make this a lot easier. } } Dave } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } ----------------- } } } } Hello listers, } } } } We are in the process of justifying, to the University, a } proposal to } } purchase an ESEM. In prepapring the business case I am } including the } } ten year financials for both FEG and Tungtsen filament } machines. Given } } that we have to break even in that time, including covering our } } depreciation, the Tungsten instrument is easier to justify } in a purely } } financial sense. } } } } From a scientific and operational viewpoint, though, I was hoping } } some } } of you could provide opinions on the relative merits of } Tungsten or FEG. } } Our current conventional SEM is a Philips XL-30 S-FEG so we } already know } } of the costs associated with the emitter. } } } } The range of samples we will be required to put in our ESEM will be } } anything you oculd possibly imagine from the Faculties of } Engineering, } } Science and Medicine. Samples will inlcude food materials } (big dairy } } industry in New Zealand), polymers, conducting polymers, mammalian } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } ceramic coatings and bees knees. We will definitely need a } hot-stage. } } } } If you would like to contact me off list with your opinions please } } feel } } free. } } } } Kind regards } } Bryony James } } Research Centre for Surface and Materials Science } } Univetrsity of Auckland } } New Zealand } } } } } } } } This incoming email to UWE has been independently scanned } for viruses } } and any virus detected has been removed using McAfee anti-virus } } software } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } } } } This email has been independently scanned for viruses and any } virus detected has been removed using McAfee anti-virus software } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:51:20 2004
Try EBSD. It will tell if you have grains or if the layer is amorphous.
gary g.
At 05:24 AM 5/18/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 09:16:24 2004
Depressing isn't it? I usually give up with cultured cells and look at them after drying with HMDS and gold coating. Have you (or anyone out there) ever taken/seen a good picture of a hydrated cultured cell at x20,000 or higher?
Dave
On Tue, 18 May 2004 08:55:19 -0500 "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote:
} In my experience medium/high magnifications for wet } specimens are limited mostly by specimens themselves and not by a } type of emitter. While I can get a nice picture of gold on carbon } at x100k, for most of my "real life" specimens magnification is } limited by x20k (or less). } } Vladimir } } } We look mostly at hydrated biological samples. We have a } } tungsten gun XL30 ESEM TMP. For looking at bacteria and } } cells a FEGESEM would be superior in terms of reduced } } damage and improved resolution. I find it hard to get } } medium/high resolution with wet biological samples. I feel } } FEGESEM would make this a lot easier. } } } } Dave } } } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } } } } } } ---------------------------------------------------------------------- } } } -------- } } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------- } } ----------------- } } } } } } Hello listers, } } } } } } We are in the process of justifying, to the University, a } } proposal to } } } purchase an ESEM. In prepapring the business case I am } } including the } } } ten year financials for both FEG and Tungtsen filament } } machines. Given } } } that we have to break even in that time, including covering our } } } depreciation, the Tungsten instrument is easier to justify } } in a purely } } } financial sense. } } } } } } From a scientific and operational viewpoint, though, I was hoping } } } some } } } of you could provide opinions on the relative merits of } } Tungsten or FEG. } } } Our current conventional SEM is a Philips XL-30 S-FEG so we } } already know } } } of the costs associated with the emitter. } } } } } } The range of samples we will be required to put in our ESEM will be } } } anything you oculd possibly imagine from the Faculties of } } Engineering, } } } Science and Medicine. Samples will inlcude food materials } } (big dairy } } } industry in New Zealand), polymers, conducting polymers, mammalian } } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } } ceramic coatings and bees knees. We will definitely need a } } hot-stage. } } } } } } If you would like to contact me off list with your opinions please } } } feel } } } free. } } } } } } Kind regards } } } Bryony James } } } Research Centre for Surface and Materials Science } } } Univetrsity of Auckland } } } New Zealand } } } } } } } } } } } } This incoming email to UWE has been independently scanned } } for viruses } } } and any virus detected has been removed using McAfee anti-virus } } } software } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } This email has been independently scanned for viruses and any } } virus detected has been removed using McAfee anti-virus software } } } } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 10:42:01 2004
When choosing a scanner for negatives, an important parameter is the Dmax (which is a log value so relatively small differences can be significant). The Dmax needed for negs is a lot more than for most printed images or documents.
At 08:55 AM 5/18/2004 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I agree that resolution is more dependent on sample than microscope. What a microscope is capable of doing with an "ideal" sample often does not relate to the real world of biological samples which are anything but ideal!
To get really good resolution you need to use higher kV values. These same values will result in the primary electrons penetrating too far into the low density biological sample resulting in substantial loss of signal in general and se1 & se2's in particular. Resulting image will be low resolution and often of much lesser quality than the same sample taken at a lower magnification.
The problem of kV can be partially overcome by using a more coherent beam with greater beam density, i.e. Field Emission. FE allows one to work at lower kV values resulting in a se yield that is still adequate while minimizing sample damage. Resulting images are likely to be higher resolution (sharper with more detail) than comparable images at similar kVs from a tungsten instrument.
However, there is a point where you go from useful magnification to "empty " magnification regardless of the gun type. This is sample dependent and must be determined accordingly. So it is unlikely that you will get any additional information from a bacteria sample at 100k than at say 50K. However you will most likely get a much sharper image at 20k with FE than you could with a W filament.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 5/18/04 8:55 AM, "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } In my experience medium/high magnifications for wet } specimens are limited mostly by specimens themselves and not by a } type of emitter. While I can get a nice picture of gold on carbon } at x100k, for most of my "real life" specimens magnification is } limited by x20k (or less). } } Vladimir } } } We look mostly at hydrated biological samples. We have a } } tungsten gun XL30 ESEM TMP. For looking at bacteria and } } cells a FEGESEM would be superior in terms of reduced } } damage and improved resolution. I find it hard to get } } medium/high resolution with wet biological samples. I feel } } FEGESEM would make this a lot easier. } } } } Dave } } } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } } } } } } ---------------------------------------------------------------------- } } } -------- } } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------- } } ----------------- } } } } } } Hello listers, } } } } } } We are in the process of justifying, to the University, a } } proposal to } } } purchase an ESEM. In prepapring the business case I am } } including the } } } ten year financials for both FEG and Tungtsen filament } } machines. Given } } } that we have to break even in that time, including covering our } } } depreciation, the Tungsten instrument is easier to justify } } in a purely } } } financial sense. } } } } } } From a scientific and operational viewpoint, though, I was hoping } } } some } } } of you could provide opinions on the relative merits of } } Tungsten or FEG. } } } Our current conventional SEM is a Philips XL-30 S-FEG so we } } already know } } } of the costs associated with the emitter. } } } } } } The range of samples we will be required to put in our ESEM will be } } } anything you oculd possibly imagine from the Faculties of } } Engineering, } } } Science and Medicine. Samples will inlcude food materials } } (big dairy } } } industry in New Zealand), polymers, conducting polymers, mammalian } } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } } ceramic coatings and bees knees. We will definitely need a } } hot-stage. } } } } } } If you would like to contact me off list with your opinions please } } } feel } } } free. } } } } } } Kind regards } } } Bryony James } } } Research Centre for Surface and Materials Science } } } Univetrsity of Auckland } } } New Zealand } } } } } } } } } } } } This incoming email to UWE has been independently scanned } } for viruses } } } and any virus detected has been removed using McAfee anti-virus } } } software } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } This email has been independently scanned for viruses and any } } virus detected has been removed using McAfee anti-virus software } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 11:19:50 2004
Gene P. Young Sr. Analytical Technologist Analytical Sciences, Polymer Characterization
The Dow Chemical Company 2301 N. Brazosport Blvd., B-1470 Freeport, Texas 77541-3257 Fax: (979) 238-0095
*(979) 238-1579 * gpyoung-at-dow.com
**************************************** Original message:
Hello Listers,
I wanted to see if anyone has had any experience scanning negatives with the Canon Canoscan 9900F. I have a low end Canoscan that I use for documents and the occasional photograph, which has given me very good results, but is not set up for negatives. I have seen the recent posts about the Epson scanner, but I am trying to keep it under a $1000 so that I do not have to treat it as a capital purchase. I have tried for the last two years to get a Coolscan 8000 on the capital budget with no luck, and do not see any scanner above $1000 as an option right now.
Thanks, Steve
Steven Lee Chief Technologists Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 750246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 02:44:19 2004
} } IbPRIA'2005 } } 2nd Iberian Conference on Pattern Recognition and Image Analysis } } June 7-9, 2005 - Estoril, Portugal } } http://ibpria2005.isr.ist.utl.pt } } } ================= } ABOUT IbPRIA'2005 } ================= } IbPRIA'2005 is the second edition of the Iberian Conference on Pattern } Recognition and Image Analysis and will be held in Estoril (Portugal) } in June 7-9, 2005. } } IbPRIA is an international event co-organised every two years, by the } Spanish and Portuguese Associations for Pattern Recognition (AERFAI } and APRP). Its mission is to bring together researchers from all over } the world working in Pattern Recognition and in all areas of Video, } Image and Signal Processing. Participants will have the opportunity to } meet colleagues working in these areas, to attend oral sessions and to } exchange ideas during poster sessions. } } IbPRIA'2005 is sponsored by IAPR-International Association for Pattern } Recognition and the conference proceedings are planned to be published } by Springer in LNCS series. } } ====== } TOPICS } ====== } The topics of the conference include, but are not limited to: } } - Pattern Recognition } - Image Analysis } - Computer Vision } - Multimedia Systems } - Statistical & Structural Pattern Recognition } - Neural Networks } - Bioinformatics } - Image Coding and Processing } - Shape and Texture Analysis } - Biomedical Pattern Analysis } - Information Systems } - Speech Recognition } - Natural Language Analysis } - Document Processing } - Robotics } - Remote Sensing } - Industrial Applications of Pattern Recognition } - Special Hardware Architectures } } ================= } PROGRAM COMMITTEE } ================= } Jake Aggarwal, University of Texas, USA } José Benedi, Polyt. University of Valencia, Spain } Isabelle Bloch, ENST, France } Hervé Bourlard, EPFL, Switzerland } Patrick Bouthemy, IRISA, France } Horst Bunke, University of Bern, Switzerland } Aurélio Campilho, University of Porto, Portugal } Gilles Celeux, Université Paris-Sud, France } Luigi Cordella, University of Napoles, Italy } Alberto Del Bimbo, University of Firenze, Italy } Hervé Delinguette, INRIA, France } Rachid Deriche, INRIA, France } José Dias, Instituto Superior Técnico, Portugal } Robert Duin, University of Delft, The Netherlands } Mário Figueiredo, Instituto Superior Técnico, Portugal } Ana Fred, Instituto Superior Técnico, Portugal } Andrew Gee, University of Cambridge, UK } Mohamed Kamel, University of Waterloo, Canada } Aggelos Katsaggelos, Northwestern University, USA } Joseph Kittler, University of Surrey, UK } Seong-Whan Lee, University of Korea, Korea } Ana Mendonça, University of Porto, Portugal } Hermann Ney, University of Aachen, Germany } Wiro Niessen, University of Utrecht, The Netherlands } Maria Petrou, University of Surrey, UK } Armando Pinho, University of Aveiro, Portugal } Ioannis Pitas, University of Tessaloniki, Greece } Filiberto Pla, University Jaume I, Spain } Richard Prager, University of Cambridge, UK } José Principe, University of Florida, USA } Ian Reid, University of Oxford, UK } Gabriella Sanniti di Baja, Istituto di Cibernética, Italy } Beatriz Santos, University of Aveiro, Portugal } Jose Santos-Victor, Inst. Superior Técnico, Portugal } Joan Serrat, Aut. University of Barcelona, Spain } Yoshiaki Shirai, Osaka University, Japan } Pierre Soille, Joint Research Centre, Italy } Karl Tombre, LORIA, France } M. Ines Torres, Univ. of the Basque Country, Spain } Emmanuele Trucco, Heriot-Watt University, UK } Alessandro Verri, University of Genova, Italy } Max Viergever, University of Utrecht, The Netherlands } Joachim Weickert, Saarland University, Germany } } ====== } CHAIRS } ====== } GENERAL CO-CHAIRS: } Jorge S. Marques (Instituto Superior Técnico, Portugal) } Nicolás Pérez de la Blanca (University of Granada, Spain) } } LOCAL CHAIR: } Pedro Pina (Instituto Superior Técnico, Portugal) } } ==================== } ORGANISING COMMITTEE } ==================== } João Sanches, João Paulo Costeira, Teresa Barata, Vitorino Ramos, } José Saraiva, Michele Mengucci, Fernando Soares } } } =================================== } PAPER SUBMISSION AND REVIEW PROCESS } =================================== } Papers should describe original and unpublished work on the topics of } the conference. Prospective authors should prepare a full paper, } written in english, not exceeding 8 pages and must submit it } electronically. } } All manuscripts will be blind-reviewed by at least two members of the } program committee. Accepted papers are intended to appear in the } Springer LNCS series which will be distributed to all participants at } the Conference. } } Submission implies that at least one of the authors has to register } and to present the communication at the conference if the paper is } accepted. } } =============== } IMPORTANT DATES } =============== } Submission of papers : Noverber 12, 2004 } Notification of acceptance: January 7, 2005 } Camera-ready : January 30, 2005 } Early registration : April 7, 2005 } Conference : June 7-9, 2005 } } ===== } VENUE } ===== } The conference will take place in Estoril, a beautiful village located } at about 20 km of Lisbon centre. } } The conference will be held at a four star hotel (Hotel Eden), that } disposes of adequate facilities for workshops and conferences. } } The international airport of Lisbon is about 40 minutes from the } venue. There is a direct bus-shuttle from/to the airport to the Hotel } every hour. Alternatively, participants can also easily reach the } venue by train, car or taxi. } } ======== } CONTACTS } ======== } IbPRIA'2005 Secretariat } CVRM / Geo-Systems Centre } Instituto Superior Técnico } Av. Rovisco Pais } 1049-001 Lisboa } PORTUGAL } } URL : http://ibpria2005.isr.ist.utl.pt } e-mail: ibpria2005-at-isr.ist.utl.pt } Tel : +351-218417438 } Fax : +351-218417442 }
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 10:22:00 2004
Members in the Greater Orlando, Florida area are cordially invited to attend a two day mini-workshop on "Cryomicrotomy and Cryoultramicrotomy for Materials Sciences."
When: Tuesday, June 8 and Wednesday, June 9, 2004, 9:00am-4:00pm
Where: Room 101, Materials Characterization Facility Advanced Materials Processing & Analysis Center University of Central Florida 12443 Research Parkway Orlando, FL 32826
What: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryomicrotomy and cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryomicrotome and cryoultramicrotome.
Background: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing" at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
Important Info: There is no charge for this workshop. Meals and refreshments will be served! Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryosectioning hands-on sessions on the second day.
Contacts: To RSVP and to reserve a spot for the hands-on sessions, please contact either Ms. Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262) or Ms. Nancy Crouch at ICMAS, Inc. ( {nancy-at-icmas.com} , 865.984.8058)
Sponsors and Organizers: University of Central Florida Advanced Materials Processing & Analysis Center Materials Characterization Facility RMC Products Group, Boeckeler Instruments, Inc.
Hope to see you there!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 10:32:23 2004
For ESEM in wet mode low voltage beam is not suitable, even at 5 kV noise level is too high.
Vladimir
} I agree that resolution is more dependent on sample than } microscope. What a microscope is capable of doing with an } "ideal" sample often does not relate to the real world of } biological samples which are anything but ideal! } } To get really good resolution you need to use higher kV } values. These same values will result in the primary } electrons penetrating too far into the low density biological } sample resulting in substantial loss of signal in general and } se1 & se2's in particular. Resulting image will be low } resolution and often of much lesser quality than the same } sample taken at a lower magnification. } } The problem of kV can be partially overcome by using a } more coherent beam with greater beam density, i.e. Field } Emission. FE allows one to work at lower kV values resulting } in a se yield that is still adequate while minimizing sample } damage. Resulting images are likely to be higher resolution } (sharper with more detail) than comparable images at similar } kVs from a tungsten instrument. } } However, there is a point where you go from useful } magnification to "empty " magnification regardless of the gun } type. This is sample dependent and must be determined } accordingly. So it is unlikely that you will get any } additional information from a bacteria sample at 100k than at } say 50K. However you will most likely get a much sharper } image at 20k with FE than you could with a W filament. } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy } } } On 5/18/04 8:55 AM, "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote: } } } } } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } -------------- } --} } - } } } } In my experience medium/high magnifications for wet specimens are } } limited mostly by specimens themselves and not by a type of } emitter. } } While I can get a nice picture of gold on carbon at x100k, } for most of } } my "real life" specimens magnification is limited by x20k (or less). } } } } Vladimir } } } } } We look mostly at hydrated biological samples. We have a tungsten } } } gun XL30 ESEM TMP. For looking at bacteria and cells a } FEGESEM would } } } be superior in terms of reduced damage and improved resolution. I } } } find it hard to get medium/high resolution with wet biological } } } samples. I feel FEGESEM would make this a lot easier. } } } } } } Dave } } } } } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } } } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } } } } } } } } } } } } --------------------------------------------------------------------- } } } - } } } } -------- } } } } The Microscopy ListServer -- Sponsor: The Microscopy } } } Society of America } } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } -------------------------------------------------------------- } } } ----------------- } } } } } } } } Hello listers, } } } } } } } } We are in the process of justifying, to the University, a } } } proposal to } } } } purchase an ESEM. In prepapring the business case I am } } } including the } } } } ten year financials for both FEG and Tungtsen filament } } } machines. Given } } } } that we have to break even in that time, including covering our } } } } depreciation, the Tungsten instrument is easier to justify } } } in a purely } } } } financial sense. } } } } } } } } From a scientific and operational viewpoint, though, I } was hoping } } } } some of you could provide opinions on the relative merits of } } } Tungsten or FEG. } } } } Our current conventional SEM is a Philips XL-30 S-FEG so we } } } already know } } } } of the costs associated with the emitter. } } } } } } } } The range of samples we will be required to put in our } ESEM will be } } } } anything you oculd possibly imagine from the Faculties of } } } Engineering, } } } } Science and Medicine. Samples will inlcude food materials } } } (big dairy } } } } industry in New Zealand), polymers, conducting polymers, } mammalian } } } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } } } ceramic coatings and bees knees. We will definitely need a } } } hot-stage. } } } } } } } } If you would like to contact me off list with your } opinions please } } } } feel free. } } } } } } } } Kind regards } } } } Bryony James } } } } Research Centre for Surface and Materials Science Univetrsity of } } } } Auckland New Zealand } } } } } } } } } } } } } } } } This incoming email to UWE has been independently scanned } } } for viruses } } } } and any virus detected has been removed using McAfee anti-virus } } } } software } } } } } } } } } } ---------------------------------------- } } } Patton, David } } } Email: David.Patton-at-uwe.ac.uk } } } "University of the West of England" } } } } } } } } } } } } This email has been independently scanned for viruses and } any virus } } } detected has been removed using McAfee anti-virus software } } } } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 14:24:44 2004
Members in the Greater Madison, Wisconsin area are cordially invited to attend a two day mini-workshop on "Cryoultramicrotomy for Materials Sciences."
When: Wednesday, June 2 and Thursday, June 3, 2004, 9:00am-4:00pm
Where: Room 374, Truax Building Madison Area Technical College 3550 Anderson Street Madison, WI 53704
(Gated lot near Truax Bldg. has audio to Parking Department. Mention this Workshop for entrance to parking lot. Enter left side of building, take main elevator to third floor)
What: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryomicrotome and cryoultramicrotome.
Background: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing" at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
Important Info: There is no charge for this workshop. Meals and refreshments will be served! Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryosectioning hands-on sessions on the second day.
Contacts: To RSVP and to reserve a spot for the hands-on sessions, please contact either Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262) or Michael Kostrna at Madison Area Technical College ( {mkostrna-at-matcmadison.edu} , 608.246.6762).
Sponsors and Organizers: Madison Area Technical College (Electron Microscopy) RMC Products Group, Boeckeler Instruments, Inc. Doug D'Arcy, Midwest Regional Representative
See you in Madison!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 17:00:14 2004
I¹d appreciate your collective wisdom and experiences for a project I¹m starting. I need to create high resolution (250 Mb minimum) photo files of current microprocessor die faces (pentium 4 type) to show the circuitry and traces with as much detail as possible. I¹d like to do this using my Nikon Optiphot and a yet to be purchased motorized positionable stage with automation software to include my Fuji S2 still camera (34 Mb sized files per image) with image stitching as an added option. I have extensive experience in Photoshop and would use that to stitch the images if the automation software package doesn¹t include that. The stage travel only needs to be about 3" (75mm) in the X and Y dimensions.
I¹ve been reading about some of the software programs available to help accomplish this. I¹ve come across Image-Pro Plus with the Scope-Pro plugin and the software program Metamorph in my Google searches. I don¹t need extensive filtering or analysis functions in the software, just the ability to create multiple, tileable images in a repeatable manner.
What economical software and hardware (such as the MultiScan-4 Low Cost Scanning Stage for example) do you have experience with in similar projects?
Do any of you have an older version of Image Pro Plus or similar software program that is sitting abandoned and alone on a shelf that needs a new home? How about a used basic scanning stage that¹s collecting dust?
Any and all advice would be appreciated. You can email me off-list if that¹s easier.
The P4 BGA devices are like PPC G3, G4 and G5 products and are flip chip BGAs. Between the inverted die and the multi layer ceramic package is a thin layer of silicone. Through this layer are many (700+) solder vias between the die and package. Once the silicone is removed (and assuming that you get the die off), the top layer greatly masks the underlying 5-8 layers (depending on part timeframe of manufacture) since it is used for interface to the ceramic package.
This top-most layer must be removed to begin to get at the actual interconnect layers. Once removed, the underlying layers can be exposed by plasma depassivation. Or, you can try wet etching.
These types of processors are face down rather than face up. This is because they are too hot compared to earlier CPUs.
The other factor is that you do not need high rez image segments to make a high rez final image. To get any reasonable image detail, you will likely need to shoot at 500X or 1000X. At these mags, your field of view is somewhere around 300u (guessing). For a 0.5cm die, you will likely take 1,000 images (guessing again). You can do the math. The point is that for a final high rez image, it is made up of many low rez image segments. The other problem you will face at high mag is the problem of low DOF. The solution is to shoot with a SEM at high tilt angle and dynamic focus. Then colorize the image.
If you need motorized and programmed stage movement, Prior E103 is a good candidate and Soft Imaging analySIS Opti is a good driving software product. Neither of these items are free.
Anyway, I have found that it is not worth the effort. But of course, YMMV.
gary g.
At 03:07 PM 5/19/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 04:11:37 2004
given the complexity of what you are doing, would it not be possible to try another method. If the die faces are flat and of the right size, why not just use a very high resolution flat bed scanner and do the whole thing in one go.
I'm sure that HP, Epson, Canon all do better than 3000dpi (optical resolution. It might certainly be worth investigating if camera photography involves a horrendous amount of image stitching (especially given the edge distortion in even the best camera). The only problems may be the reflective light quality of the scanner or how much depth of field there would be in a scanner image if the die has lots of depth to its layers.
My apologies if I've missed some technical point.
Malc
Malcolm Haswell e.m. unit University of Sunderland UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Doug Baldwin {dougbaldwin-at-mindspring.com}
My thanks to everyone who replied to my legal/ethics query. After my client became a bit more educated regarding the health risk we discovered and checked with their counsel, they will do the right thing. The twist here was that they were analyzing competitors' products which are made overseas, so there are issues of diplomacy.
Alan Stone
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 09:52:22 2004
I wish to set up a logging system to record instrument use. Ideally it would log date and time users start and finish work on a particular project on each of our instruments so that we can assess use of our facility.
At present users fill in a log book when they use an instrument. It is easy to get general data (25 sessions to the page gives an idea of instrument use but not user). I don't want to waste time transcribing the data from the 20 instrument log books into a computer. Neither do I want to spend a lot of money on each machine to gather the information, however, most machines already have a PC attached running the instrument or some ancillary equipment and these are all networked.
What systems are being used at other sites? Any suggestions of inexpensive systems that would work on two sites 6 miles apart?
Regards, Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 15:49:26 2004
Hello Everyone We have a Nonotech Semprep2 sputter coater which is so old the company no longer exists. The glass cylinder has now got one more gouge in it and it will have to lose at least 3 cm to grind it out. Does anyone have any experience with where to buy a new glass cylinder?
Many thanks Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 17:26:04 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mocherla-at-eng.fsu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, May 20, 2004 at 10:43:53 ---------------------------------------------------------------------------
I really appreciate this group for their support in my work.
I observed that for negative staining, carbon coated Formvar grid was used mostly. I would like to know if there is any significance in using this particlular grid or Can I use Carbon support film on Copper or Gold grid.
On May 20, 2004, at 3:35 PM, by way of MicroscopyListserver wrote:
} Title-Subject: [Microscopy] [Filtered] Liposome Grids } } Question: Dear authors, } } I really appreciate this group for their support in my work. } } I observed that for negative staining, carbon coated Formvar grid was } used mostly. I would like to know if there is any significance in } using this particlular grid or Can I use Carbon support film on Copper } or Gold grid. } Dear Supriya, The grids most often used for negative staining are copper grids on which there is a layer of formvar and usually an additional layer of carbon. These are often referred to as carbon-formvar grids or carbon-coated formvar grids. It is much easier to use a carbon-formvar support film than to use just a carbon film for support. The resulting film is much stronger than plain carbon, and does not degrade the images of negatively stained preps. The web sites of several of the EM supply companies describe their carbon-formvar grids. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 20:04:13 2004
Interestingly, I had the same thought and tried this technique. I stopped by one of the area's premier graphics arts service bureaus with an 8000+ dpi flat bed scanner. We put the open faced CPU chip on the glass and proceeded to scan it. It was awful. The main problem was that the light source on the scanner was off-axis to the scanning array. This renders the face of the chip dark and worthless.
Photos of microprocessors benefit from coaxial lighting since the metal parts and traces reflect the light straight back to the lens and light source. Lighting that's even slightly off-axis shows a marked drop off in detail and reflectivity of the die faces.
The high dpi flat bed scanners are set up for flat artwork such as paintings and drawings where side lighting is fine for rendering all the detail but not for reflective metal parts.
Doug
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Doug } } given the complexity of what you are doing, would it not be possible to try } another method. If the die faces are flat and of the right size, why not just } use a very high resolution flat bed scanner and do the whole thing in one go. } } I'm sure that HP, Epson, Canon all do better than 3000dpi (optical resolution. } It might certainly be worth investigating if camera photography involves a } horrendous amount of image stitching (especially given the edge distortion in } even the best camera). The only problems may be the reflective light quality } of the scanner or how much depth of field there would be in a scanner image if } the die has lots of depth to its layers. } } My apologies if I've missed some technical point. } } Malc } } Malcolm Haswell } e.m. unit } University of Sunderland } UK } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } ----- Original Message ----- } } From: Doug Baldwin {dougbaldwin-at-mindspring.com} } Date: Wednesday, May 19, 2004 11:07 pm } Subject: [Microscopy] Hi-Res Microprocessor Photos } } } } } } } ------------------------------------------------------------------- } } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } AmericaTo Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } ------------------------------------------------------------------- } } ---- } } } } Hello Fellow List Members, } } } } Id appreciate your collective wisdom and experiences for a } } project Im } } starting. I need to create high resolution (250 Mb minimum) photo } } files of } } current microprocessor die faces (pentium 4 type) to show the } } circuitry and } } traces with as much detail as possible. Id like to do this using } } my Nikon } } Optiphot and a yet to be purchased motorized positionable stage with } } automation software to include my Fuji S2 still camera (34 Mb } } sized files } } per image) with image stitching as an added option. I have extensive } } experience in Photoshop and would use that to stitch the images if the } } automation software package doesnt include that. The stage travel } } onlyneeds to be about 3" (75mm) in the X and Y dimensions. } } } } Ive been reading about some of the software programs available to } } helpaccomplish this. Ive come across Image-Pro Plus with the } } Scope-Pro plugin } } and the software program Metamorph in my Google searches. I dont need } } extensive filtering or analysis functions in the software, just } } the ability } } to create multiple, tileable images in a repeatable manner. } } } } What economical software and hardware (such as the MultiScan-4 Low } } CostScanning Stage for example) do you have experience with in } } similar projects? } } } } Do any of you have an older version of Image Pro Plus or similar } } softwareprogram that is sitting abandoned and alone on a shelf } } that needs a new } } home? How about a used basic scanning stage thats collecting dust? } } } } Any and all advice would be appreciated. You can email me off-list } } if thats } } easier. } } } } Thanks, } } Doug Baldwin } } Baldwin Hi-Tech Photography } } dougbaldwin-at-mindspring.com } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 22:16:30 2004
There is a photographer here in Oz who produces absolutely brilliant images by scanning objects on a flat-bed scanner. They are very high resolution, look as good as the images of leaves, flowers, insects (even very shiny ones) we get using a good digital camera - Olympus DP70 or ProgRes or similar, on a dissecting microscope. He's developed some ways of using the scanner to get such great images. Name is Stuart Owen Fox, go here http://www.smartstate.qld.gov.au/images/icase4g_2.jpg for an example. cheers, Rosemary
} From: Doug Baldwin {dougbaldwin-at-mindspring.com} } Date: Thu, 20 May 2004 18:10:14 -0700 } To: {microscopy-at-msa.microscopy.com} } Cc: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} } Subject: [Microscopy] Re: Hi-Res Microprocessor Photos } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Malcolm, } } Interestingly, I had the same thought and tried this technique. I stopped by } one of the area's premier graphics arts service bureaus with an 8000+ dpi } flat bed scanner. We put the open faced CPU chip on the glass and proceeded } to scan it. It was awful. The main problem was that the light source on the } scanner was off-axis to the scanning array. This renders the face of the } chip dark and worthless. } } Photos of microprocessors benefit from coaxial lighting since the metal } parts and traces reflect the light straight back to the lens and light } source. Lighting that's even slightly off-axis shows a marked drop off in } detail and reflectivity of the die faces. } } The high dpi flat bed scanners are set up for flat artwork such as paintings } and drawings where side lighting is fine for rendering all the detail but } not for reflective metal parts. } } Doug } } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } --} - } } } } Doug } } } } given the complexity of what you are doing, would it not be possible to try } } another method. If the die faces are flat and of the right size, why not just } } use a very high resolution flat bed scanner and do the whole thing in one go. } } } } I'm sure that HP, Epson, Canon all do better than 3000dpi (optical } } resolution. } } It might certainly be worth investigating if camera photography involves a } } horrendous amount of image stitching (especially given the edge distortion in } } even the best camera). The only problems may be the reflective light quality } } of the scanner or how much depth of field there would be in a scanner image } } if } } the die has lots of depth to its layers. } } } } My apologies if I've missed some technical point. } } } } Malc } } } } Malcolm Haswell } } e.m. unit } } University of Sunderland } } UK } } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } } } } } } ----- Original Message ----- } } } From: Doug Baldwin {dougbaldwin-at-mindspring.com} } } Date: Wednesday, May 19, 2004 11:07 pm } } Subject: [Microscopy] Hi-Res Microprocessor Photos } } } } } } } } } } } ------------------------------------------------------------------- } } } ----------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } AmericaTo Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } } ------------------------------------------------------------------- } } } ---- } } } } } } Hello Fellow List Members, } } } } } } Id appreciate your collective wisdom and experiences for a } } } project Im } } } starting. I need to create high resolution (250 Mb minimum) photo } } } files of } } } current microprocessor die faces (pentium 4 type) to show the } } } circuitry and } } } traces with as much detail as possible. Id like to do this using } } } my Nikon } } } Optiphot and a yet to be purchased motorized positionable stage with } } } automation software to include my Fuji S2 still camera (34 Mb } } } sized files } } } per image) with image stitching as an added option. I have extensive } } } experience in Photoshop and would use that to stitch the images if the } } } automation software package doesnt include that. The stage travel } } } onlyneeds to be about 3" (75mm) in the X and Y dimensions. } } } } } } Ive been reading about some of the software programs available to } } } helpaccomplish this. Ive come across Image-Pro Plus with the } } } Scope-Pro plugin } } } and the software program Metamorph in my Google searches. I dont need } } } extensive filtering or analysis functions in the software, just } } } the ability } } } to create multiple, tileable images in a repeatable manner. } } } } } } What economical software and hardware (such as the MultiScan-4 Low } } } CostScanning Stage for example) do you have experience with in } } } similar projects? } } } } } } Do any of you have an older version of Image Pro Plus or similar } } } softwareprogram that is sitting abandoned and alone on a shelf } } } that needs a new } } } home? How about a used basic scanning stage thats collecting dust? } } } } } } Any and all advice would be appreciated. You can email me off-list } } } if thats } } } easier. } } } } } } Thanks, } } } Doug Baldwin } } } Baldwin Hi-Tech Photography } } } dougbaldwin-at-mindspring.com } } } } } } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 23:52:06 2004
A good glassblower can sort it. Either by producing another or taking the gouge out. I have done that in the past on a VERY OLD Edwards. Definitely cheaper than the scientific company's. The quality of the work was good and the new dome is still in operation after 6 years.
-----Original Message----- } From: Elaine Humphrey [mailto:ech-at-interchange.ubc.ca] Sent: Thursday, May 20, 2004 10:59 PM To: microscopy-at-msa.microscopy.com
Hello Everyone We have a Nonotech Semprep2 sputter coater which is so old the company no longer exists. The glass cylinder has now got one more gouge in it and it will have to lose at least 3 cm to grind it out. Does anyone have any experience with where to buy a new glass cylinder?
Many thanks Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 03:55:22 2004
Regarding stitching images, we have successfully stitched micrographs using PanaVue ImageAssembler (www.panavue.com) as it works with flat perspective. Have not tried to stitch very large numbers of images though.
With regards Thor
Dr Thor Bostrom Analytical EM Facility; and School of Physical and Chemical Sciences, Queensland University of Technology (QUT) Brisbane, QLD 4001, Australia
does anybody know how is the phase in AFM software Nanoscope III for Dimension 2100 defined? Could the bright regions in the phase image be assigned to the depressions, if we are working in the attractive force regime? Thank's a lot for any hint.
I seriously doubt there is one as to unambiguously identify fibrous asbestos is the job of TEM and (not or) an EDS on TEM.
**************************************** Chaoying Ni, PhD The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-8354 (O); -2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
On Fri, 21 May 2004, Allan Mitchell wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi all } } Can anyone suggest a good text or reference for helping to identify } asbestos bodies in lung using an SEM, and using EDS. } } Many thanks } } Regards } } Allan }
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 07:57:16 2004
I have some cells in solution. Is there some general technique to preserve them so that I can put some down on a carbon membrane coated TEM grid without them shriveling up? Can anyone refer me to a good reference on TEM sample prep of cells?
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 08:09:47 2004
The Mineralogical Society of America has a series of books on mineralogical and geochemical topics. Volume 28 is titled "Health Effects of Mineral Dusts" and covers a wide range of topics, from basic asbestos mineralogy to cellular and molecular mechanisms for disease. I'm not sure if it specifically deals with identifying asbestos bodies in lung tissue specificaly but it should have some references, and there should be useful information to help with EDS identification. It's available at the MSA (the other MSA) website at www.minsocam.org/MSA/RIM/ for a very reasonable price for the amount of information contained ($32 US).
Best Regards, Bryan Bandli
Allan Mitchell wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi all } } Can anyone suggest a good text or reference for helping to identify } asbestos bodies in lung using an SEM, and using EDS. } } Many thanks } } Regards } } Allan } }
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From MicroscopyL-request-at-ns.microscopy.com Fri May 21 10:48:40 2004
I had a request recently to perform EDS on a presumed crysotile crystal/shard that had been located with TEM on a grid. We have an Oxford INCA system on our FEI Quanta 400, so I resurrected an old grid holder from a salvaged RCA EMU4, mounted the grid in holder in the ESEM, located the offending grid square and performed a point and ID on the crystal in question.
As Chaoying suggests, this would not be in concert with NIOSH 7402 (TEM) http://www.cdc.gov/niosh/nmam/pdfs/7402.pdf, or NIOSH 9000 (XRD) http://www.cdc.gov/niosh/nmam/pdfs/7402.pdf, identifications of asbestos, to give quick examples.
Other sources and possibilities for your problem(?): SEM+: http://sup.ultrakohl.com/News-nov/mesoth.htm
EBSD:
http://www.asbestostemlabs.com/staff.htm
SEM/EBSD http://www.dxcicdd.com/01/pdf/D-075.pdf
I don't believe that EBSD/SEM has been approved for asbestos:
But that doesn't mean that SEM can't be used in simple identifications of morphology and composition. The real problem is to find it, if you are looking in tissue. Mapping for the relevant elements should work, but for very small crystals, you will be working at or above the 'good result' window for EDS and mapping when you want to identify and image at high mag.
Hope some of this helps,
Fred
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson-at-wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/
-----Original Message----- } From: Allan Mitchell [mailto:allan.mitchell-at-stonebow.otago.ac.nz] Sent: Friday, May 21, 2004 1:01 AM To: microscopy-at-msa.microscopy.com
Hi all
Can anyone suggest a good text or reference for helping to identify asbestos bodies in lung using an SEM, and using EDS.
Many thanks
Regards
Allan
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 11:05:04 2004
Do you want to just look at them whole, in a HVEM? No embedding/sectioning? What do you want to see?
On Fri, 21 May 2004, Larsen, Michael (Research) wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I have some cells in solution. Is there some general technique to preserve } them so that I can put some down on a carbon membrane coated TEM grid } without them shriveling up? Can anyone refer me to a good reference on TEM } sample prep of cells? } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 11:51:52 2004
Today's microprocessors have micron scale metal lines. You would require submicron resolution to distinguish the lines. A flat bed scanner that would resolve micron scale lines would need at least 25,000 DPI resolution just to detect the lines.
John Mardinly Intel
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-csiro.au] Sent: Thursday, May 20, 2004 8:24 PM To: Doug Baldwin; microscopy-at-msa.microscopy.com Cc: Malcolm Haswell
There is a photographer here in Oz who produces absolutely brilliant images by scanning objects on a flat-bed scanner. They are very high resolution, look as good as the images of leaves, flowers, insects (even very shiny ones) we get using a good digital camera - Olympus DP70 or ProgRes or similar, on a dissecting microscope. He's developed some ways of using the scanner to get such great images. Name is Stuart Owen Fox, go here http://www.smartstate.qld.gov.au/images/icase4g_2.jpg for an example. cheers, Rosemary
} From: Doug Baldwin {dougbaldwin-at-mindspring.com} } Date: Thu, 20 May 2004 18:10:14 -0700 } To: {microscopy-at-msa.microscopy.com} } Cc: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} } Subject: [Microscopy] Re: Hi-Res Microprocessor Photos } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Malcolm, } } Interestingly, I had the same thought and tried this technique. I stopped by } one of the area's premier graphics arts service bureaus with an 8000+ dpi } flat bed scanner. We put the open faced CPU chip on the glass and proceeded } to scan it. It was awful. The main problem was that the light source on the } scanner was off-axis to the scanning array. This renders the face of the } chip dark and worthless. } } Photos of microprocessors benefit from coaxial lighting since the metal } parts and traces reflect the light straight back to the lens and light } source. Lighting that's even slightly off-axis shows a marked drop off in } detail and reflectivity of the die faces. } } The high dpi flat bed scanners are set up for flat artwork such as paintings } and drawings where side lighting is fine for rendering all the detail but } not for reflective metal parts. } } Doug } } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } --} - } } } } Doug } } } } given the complexity of what you are doing, would it not be possible to try } } another method. If the die faces are flat and of the right size, why not just } } use a very high resolution flat bed scanner and do the whole thing in one go. } } } } I'm sure that HP, Epson, Canon all do better than 3000dpi (optical } } resolution. } } It might certainly be worth investigating if camera photography involves a } } horrendous amount of image stitching (especially given the edge distortion in } } even the best camera). The only problems may be the reflective light quality } } of the scanner or how much depth of field there would be in a scanner image } } if } } the die has lots of depth to its layers. } } } } My apologies if I've missed some technical point. } } } } Malc } } } } Malcolm Haswell } } e.m. unit } } University of Sunderland } } UK } } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } } } } } } ----- Original Message ----- } } } From: Doug Baldwin {dougbaldwin-at-mindspring.com} } } Date: Wednesday, May 19, 2004 11:07 pm } } Subject: [Microscopy] Hi-Res Microprocessor Photos } } } } } } } } } } } ------------------------------------------------------------------- } } } ----------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } AmericaTo Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } } ------------------------------------------------------------------- } } } ---- } } } } } } Hello Fellow List Members, } } } } } } Id appreciate your collective wisdom and experiences for a } } } project Im } } } starting. I need to create high resolution (250 Mb minimum) photo } } } files of } } } current microprocessor die faces (pentium 4 type) to show the } } } circuitry and } } } traces with as much detail as possible. Id like to do this using } } } my Nikon } } } Optiphot and a yet to be purchased motorized positionable stage with } } } automation software to include my Fuji S2 still camera (34 Mb } } } sized files } } } per image) with image stitching as an added option. I have extensive } } } experience in Photoshop and would use that to stitch the images if the } } } automation software package doesnt include that. The stage travel } } } onlyneeds to be about 3" (75mm) in the X and Y dimensions. } } } } } } Ive been reading about some of the software programs available to } } } helpaccomplish this. Ive come across Image-Pro Plus with the } } } Scope-Pro plugin } } } and the software program Metamorph in my Google searches. I dont need } } } extensive filtering or analysis functions in the software, just } } } the ability } } } to create multiple, tileable images in a repeatable manner. } } } } } } What economical software and hardware (such as the MultiScan-4 Low } } } CostScanning Stage for example) do you have experience with in } } } similar projects? } } } } } } Do any of you have an older version of Image Pro Plus or similar } } } softwareprogram that is sitting abandoned and alone on a shelf } } } that needs a new } } } home? How about a used basic scanning stage thats collecting dust? } } } } } } Any and all advice would be appreciated. You can email me off-list } } } if thats } } } easier. } } } } } } Thanks, } } } Doug Baldwin } } } Baldwin Hi-Tech Photography } } } dougbaldwin-at-mindspring.com } } } } } } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 14:17:27 2004
I'll try again. Does anyone have a Gatan model 626-300 cryoholder to sell or donate to a university? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 14:21:00 2004
Image-Pro Plus is a very good image analysis software with extensive capabilities. But there is a another product from Media Cybernetics which is "Image-Pro Discovery". This is the smaller version of Image-Pro Plus and therefore cheaper. It allows you to capture X-Y automatically in combination with Scope-Pro. The software gives you the possibility to select the order how you want to capture the images and it stitches them automatically. You also could capture Z-stacks automatically either by moving the microscope stage or with a motorized objective (Piezo) which gives you very precise results.
However if you want to work with an automatic x-y stage you should work with a camera which is integrated in the capturing software because then the software also captures the images, then the stage moves on, the image is captured and so on. With your Fuji camera you have to do the capturing manually which is very cumbersome. You should use a digital CCD camera which is connected to your computer and shows you the live image on the computer monitor.
DB} ------------------------------------------------------------------------------ DB} The Microscopy ListServer -- Sponsor: The Microscopy Society of America DB} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver DB} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html DB} -------------------------------------------------------------------------------
DB} Hello Fellow List Members,
DB} I¹d appreciate your collective wisdom and experiences for a project I¹m DB} starting. I need to create high resolution (250 Mb minimum) photo files of DB} current microprocessor die faces (pentium 4 type) to show the circuitry and DB} traces with as much detail as possible. I¹d like to do this using my Nikon DB} Optiphot and a yet to be purchased motorized positionable stage with DB} automation software to include my Fuji S2 still camera (34 Mb sized files DB} per image) with image stitching as an added option. I have extensive DB} experience in Photoshop and would use that to stitch the images if the DB} automation software package doesn¹t include that. The stage travel only DB} needs to be about 3" (75mm) in the X and Y dimensions.
DB} I¹ve been reading about some of the software programs available to help DB} accomplish this. I¹ve come across Image-Pro Plus with the Scope-Pro plugin DB} and the software program Metamorph in my Google searches. I don¹t need DB} extensive filtering or analysis functions in the software, just the ability DB} to create multiple, tileable images in a repeatable manner.
DB} What economical software and hardware (such as the MultiScan-4 Low Cost DB} Scanning Stage for example) do you have experience with in similar projects?
DB} Do any of you have an older version of Image Pro Plus or similar software DB} program that is sitting abandoned and alone on a shelf that needs a new DB} home? How about a used basic scanning stage that¹s collecting dust?
DB} Any and all advice would be appreciated. You can email me off-list if that¹s DB} easier.
Bill I disagree with you: carbon coated Formvar film do degrade the image in your words versus just carbon. Formvar film is quite thick in compare with "plain carbon", so it "adsorbs" more electrons and therefore "degrade the image"... Another problem with Formvar and other plastic support films is that they change their properties under the beam which cause the image drift... So, in my point of view, it's just wrong statement that carbon coated Formvar film "does not degrade the images of negatively stained preps". It's quite opposite: pure thin carbon film is very stable under the beam, much thinner than any plastic film and therefore provides best possible (for biological samples) support material in terms of stability and "transparency" (to the electrons) for negative staining. Carbon coated plastic support films are popular just because it's much easier to produce in the Lab (and not necessary the best choice). In my Lab we routinely use 1.4-1.8 nm thick "pure" carbon support films over "holey" film. You may not use such thin film for huge objects like whole cell, but it works great for macromolecules of any sort. "Carbon over holey film" grids are available thorough major EM suppliers. Interestingly, I never used carbon coated Formvar films. I used to use carbon coating on cellulose-derivative films like Parlodion (because it drifted so much without carbon) and I used "naked" Formvar film because it's more stable under the beam than other variety of films. Formvar film looks much "darker" in the scope than Parlodion, so I prefer to use carbon coated Parlodion film if I could not use "normal carbon": very contaminated samples with a lot of aggregates, cells etc. Someone may need to try "naked" Parlodion (or similar) because good hydrophilic properties of cellulose-deviated films if Formvar of carbon is too hydrophobic to them (at the huge price of film instability in the beam). I don't like glow-discharge, personally: I would rather use poly-lysine coating. Have a great weekend, Sergey
At 05:39 PM 5/20/2004 -0700, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri May 21 16:18:31 2004
Hi listers I have a problem I am hoping that someone out there can help me with. 7 years ago our lab jointly bought a Photometrix SenSys PVCAM with another lab. The labs are splitting up. We need to determine the current value of the camera so that one lab can pay the other lab. Can anybody point me to this data? Thanx David
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 17:29:47 2004
We have been using the Epson Perfection 3200 scanner for 8x10 cm TEM negatives for about a year now, and find it excellent. The resolution (3200 dpi optical) is almost as good as the 4000 dpi on the Nikon 8000 etc., and we successfully scanned some very dense (accidentally overexposed) negatives when our TEM's exposure meter went on the fritz. We mount the negatives in the standard 4x5" negative carrier. (The Nikon negative holder doesn't fit 8x10 cm negatives without modification, or cutting the negatives down to size.) The 3200 was listed at $299 on the Epson website the last time I looked.
Cheers, Tom Kosel
Original message:
Hello Listers,
I wanted to see if anyone has had any experience scanning negatives with the Canon Canoscan 9900F. I have a low end Canoscan that I use for documents and the occasional photograph, which has given me very good results, but is not set up for negatives. I have seen the recent posts about the Epson scanner, but I am trying to keep it under a $1000 so that I do not have to treat it as a capital purchase. I have tried for the last two years to get a Coolscan 8000 on the capital budget with no luck, and do not see any scanner above $1000 as an option right now.
Thanks, Steve
Steven Lee Chief Technologists Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 750246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu -- ********************************* Thomas H. Kosel Department of Electrical Engineering University of Notre Dame Notre Dame, IN 46556 (574) 631-5642 (201 Cushing) (574) 631-4393 FAX kosel-at-nd.edu *********************************
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 19:08:23 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
"Electron Microscopy of Thin Crystals" Hirsch, Howie, Nicholson, Pashley & Whelan Pub: Robert E. Krieger ISBN: 0-88275-376-2
"Transmission Electron Microscopy of Materials" Thomas & Goringe Pub: John Wiley & Sons ISBN: 0-471-12244-0
"Defect Analysis in Electron Microscopy" Loretto & Smallman Pub: Chapman & Hall ISBN: 0-470-54760-X
The latter, in particular, is quite detailed about dislocation analysis with specific instructions. It also gives details of weak-beam methods, which give much higher resolution images of dislocations, very useful if you have closely packed dislocations.
Unfortunately, I suspect all are no longer in print. -- Larry Stoter PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail will automatically be deleted. :-)
From MicroscopyL-request-at-ns.microscopy.com Sat May 22 04:58:19 2004
When I read literatures, I have been all the time confused by the resolution definition in transmission electron microscopy. Could you please kindly help?
For a FEG-HRTEM, what is the "line resolution" as opposed to the "information resolution"? What is the difference between each other?
How could I achieve the "line resolution" and the "information resolution" in TEM experiments? And, how can I resolve (identify, or "see") the "line resolution" and the "information resolution" respectively from a state-of-the-art HREM image?
Thank you for your time. Wish you a nice weekend,
-Juha
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From MicroscopyL-request-at-ns.microscopy.com Sat May 22 09:13:06 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (antuni-at-aol.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 21, 2004 at 16:19:26 ---------------------------------------------------------------------------
Email: antuni-at-aol.com Name: Anthony Ribaudo
Organization: Consultant
Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical Microscopy
Question: Can one identify asbestos only utilizing optical microscopy without the aid of electron diffraction/TEM ?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (christian_b_a-at-hotmail.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, May 21, 2004 at 09:07:25 ---------------------------------------------------------------------------
Email: christian_b_a-at-hotmail.com Name: christian albano
Organization: Neuropsychiatric Research Institute
Education: Graduate College
Location: Fargo, ND, USA
Question: Does using the Fluoview Olympus LSCM for non-floresence degrade the equipment lifetime?
I remember an edition of "This Old House" or one of those types of shows where they took some shingle samples to a lab for testing.
The lab used polarized microscopy to do at least an "initial" screening and determine that the shingles "most likely" contained asbestos.
The "would do some additional testing" to verify (it was, and the house needed to have a tent built around it, etc., etc.). I assume the additional testing was something along the lines of TEM, etc.
The conclusion I drew was that if you had materials that you were expecting asbestos, this (polarized examination) was a good screening tool. If the fibers didn't change from one specific color to another as the analyzer was rotated, you didn't have asbestos. If they did, you probably have asbestos, but there are other fibers that may do the same thing.
These were also fibers, not dust, and I don't know if that would affect the results or not.
Sorry if this wasn't useful.
John R.
-----Original Message----- } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] Sent: Saturday, May 22, 2004 9:23 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (antuni-at-aol.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 21, 2004 at 16:19:26 ------------------------------------------------------------------------ ---
Email: antuni-at-aol.com Name: Anthony Ribaudo
Organization: Consultant
Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical Microscopy
Question: Can one identify asbestos only utilizing optical microscopy without the aid of electron diffraction/TEM ?
Only one of the forms of asbestos is harmful to humans. Chrysotile and then only if it is inhaled and the particle size is such it lodges in the lung so it can form a site for cancer to get a start. Chrysotile is only a small percentage of asbestos and the particle size must fall in the 5 to 15 micron range to stay in the lungs.
These conditions are almost only found in asbestos worker and miners and not in the general public. In the case of high exposure to asbestos cigarette smoking is also a very large contributor to the cancer caused by asbestos.
Had the asbestos studies been allowed to be fully peer reviewed before legislation was passed we would probably still be using asbestos in many applications today with more protection for asbestos workers and minors and restrictions on some products such as asbestos powder and asbestos cloth. Unfortunately the legislators won't review thier laws once they are passed and the western world is spending needless billions of dollars in asbestos abatement in every old building that is renovated.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
} From: "Chiphead" {chiphead-at-sbcglobal.net}
: : Warning: Non scientific answer: : : I remember an edition of "This Old House" or one of those types of shows : where they took some shingle samples to a lab for testing. : : The lab used polarized microscopy to do at least an "initial" screening : and determine that the shingles "most likely" contained asbestos. : : The "would do some additional testing" to verify (it was, and the house : needed to have a tent built around it, etc., etc.). I assume the : additional testing was something along the lines of TEM, etc. : : The conclusion I drew was that if you had materials that you were : expecting asbestos, this (polarized examination) was a good screening : tool. If the fibers didn't change from one specific color to another as : the analyzer was rotated, you didn't have asbestos. If they did, you : probably have asbestos, but there are other fibers that may do the same : thing. : : These were also fibers, not dust, and I don't know if that would affect : the results or not. : : Sorry if this wasn't useful. : : John R. : : -----Original Message----- : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] : Sent: Saturday, May 22, 2004 9:23 AM : To: microscopy-at-ns.microscopy.com : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical : Microscopy : : : : ------------------------------------------------------------------ ------ : ------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------ : ------- : : Below is the result of your feedback form (NJZFM-ultra-55). It was : submitted by (antuni-at-aol.com) from : http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, : May 21, 2004 at 16:19:26 : ------------------------------------------------------------------ ------ : --- : : Email: antuni-at-aol.com : Name: Anthony Ribaudo : : Organization: Consultant : : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical : Microscopy : : Question: Can one identify asbestos only utilizing optical microscopy : without the aid of electron diffraction/TEM ? : : : Anthony Ribaudo : : : : ------------------------------------------------------------------ ------ : --- : : : : :
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 09:31:26 2004
This is off the topic of identifying asbestos, but thought I should comment that just yesterday there was a program on the radio (streaming audio available at http://www.abc.net.au/rn/talks/bbing/, transcript available by Thursday) about new cases of asbestosis and mesothelioma showing up. The young people (20s and 30s) now getting this lived in high asbestos areas as kids, were children of builders (and builders themselves) who demolished or renovated fibro houses, etc. Perhaps just an Australian problem.... Rosemary
} From: "Gordon Couger" {gcouger-at-provalue.net} } Date: Sat, 22 May 2004 16:09:06 -0500 } To: "Chiphead" {chiphead-at-sbcglobal.net} , "'by way of MicroscopyListserver'" } {antuni-at-aol.com} , {microscopy-at-ns.microscopy.com} } Subject: [Microscopy] Re: RE: viaWWW: Identifying Asbestos Using ptical } Microscopy } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Only one of the forms of asbestos is harmful to humans. Chrysotile } and then only if it is inhaled and the particle size is such it } lodges in the lung so it can form a site for cancer to get a start. } Chrysotile is only a small percentage of asbestos and the particle } size must fall in the 5 to 15 micron range to stay in the lungs. } } These conditions are almost only found in asbestos worker and miners } and not in the general public. In the case of high exposure to } asbestos cigarette smoking is also a very large contributor to the } cancer caused by asbestos. } } Had the asbestos studies been allowed to be fully peer reviewed } before legislation was passed we would probably still be using } asbestos in many applications today with more protection for } asbestos workers and minors and restrictions on some products such } as asbestos powder and asbestos cloth. Unfortunately the legislators } won't review thier laws once they are passed and the western world } is spending needless billions of dollars in asbestos abatement in } every old building that is renovated. } } Gordon } Gordon Couger gcc-at-couger.com } } I collect links on information related to light microscopes. } http://www.couger.com/microscope/links/gclinks.html } Please forward any links or information you think might be useful to } others. } Microscope Manual at www.science-info.org } } } From: "Chiphead" {chiphead-at-sbcglobal.net} } } : } : Warning: Non scientific answer: } : } : I remember an edition of "This Old House" or one of those types of } shows } : where they took some shingle samples to a lab for testing. } : } : The lab used polarized microscopy to do at least an "initial" } screening } : and determine that the shingles "most likely" contained asbestos. } : } : The "would do some additional testing" to verify (it was, and the } house } : needed to have a tent built around it, etc., etc.). I assume the } : additional testing was something along the lines of TEM, etc. } : } : The conclusion I drew was that if you had materials that you were } : expecting asbestos, this (polarized examination) was a good } screening } : tool. If the fibers didn't change from one specific color to } another as } : the analyzer was rotated, you didn't have asbestos. If they did, } you } : probably have asbestos, but there are other fibers that may do the } same } : thing. } : } : These were also fibers, not dust, and I don't know if that would } affect } : the results or not. } : } : Sorry if this wasn't useful. } : } : John R. } : } : -----Original Message----- } : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] } : Sent: Saturday, May 22, 2004 9:23 AM } : To: microscopy-at-ns.microscopy.com } : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical } : Microscopy } : } : } : } : ------------------------------------------------------------------ } ------ } : ------ } : The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } : To Subscribe/Unsubscribe -- } : http://www.msa.microscopy.com/MicroscopyListserver } : On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } : ------------------------------------------------------------------ } ------ } : ------- } : } : Below is the result of your feedback form (NJZFM-ultra-55). It } was } : submitted by (antuni-at-aol.com) from } : http://microscopy.com/MicroscopyListserver/MLFormMail.html on } Friday, } : May 21, 2004 at 16:19:26 } : ------------------------------------------------------------------ } ------ } : --- } : } : Email: antuni-at-aol.com } : Name: Anthony Ribaudo } : } : Organization: Consultant } : } : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos } Using ptical } : Microscopy } : } : Question: Can one identify asbestos only utilizing optical } microscopy } : without the aid of electron diffraction/TEM ? } : } : } : Anthony Ribaudo } : } : } : } : ------------------------------------------------------------------ } ------ } : --- } : } : } : } : } : } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 09:50:34 2004
Sorry, I forgot to state that the cryoholder we're looking for would be for a Hitachi S-900 in-lens FESEM, which uses a Hitachi TEM holder.
Phil -- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 11:14:18 2004
It is hard to believe other forms of asbestos: amosite asbestos(grunerite), actinolite-tremolite asbestos, anthophyllite (asbestos)and crocidolite asbestos(riebeckite) are not harmful. Have any studies been done on this? Chrysotile asbestos makes up to +90% of all asbestos in North America. (please correct if somebody has published figures on this).
Asbestos has been linked to mesothelioma and, also, asbestos is the cause of asbestosis(also harmful to humans). Isn't it?
I agree. If asbestos studies had been fully reviewed we would still using asbestos it has unique chemical and physical properties, especially for a mineral. The politicians used asbestos as a political football first scaring the public then passing legislation covering asbestos in schools, AHERA. Yes many problems have occurred as a result of this. The regulation should be reviewed and changed(if if hasn't already been).
Yes, asbestos can be identified by polarized light microscopy, not OLM or PCM. Using dispersion staining techniques to determine refractive indices, a properly prepared asbestos fiber can be identified, by PLM. Other measured properties: pleochroism, sign of elongation, and angle of extinction plus physical characteristic further aid the PLM analyst in the proper identification. Preparation of the fiber and proper training of the analyst are the critical factors here. Binder/matrix effects may hinder the analysis, for example chrysotile in vinyl floor tile.
Where are all the McCrone(ies) on this? Maybe this subject has been hashed out in the past. If so pardon me for the repeated redundancy.
Regards,
Timothy J. Bard, Scientist Scanning Electron Microscopy OSRAM SYLVANIA Towanda, PA
U
-----Original Message----- } From: Gordon Couger [mailto:gcouger-at-provalue.net] Sent: Saturday, May 22, 2004 5:09 PM To: Chiphead; 'by way of MicroscopyListserver'; microscopy-at-ns.microscopy.com
Only one of the forms of asbestos is harmful to humans. Chrysotile and then only if it is inhaled and the particle size is such it lodges in the lung so it can form a site for cancer to get a start. Chrysotile is only a small percentage of asbestos and the particle size must fall in the 5 to 15 micron range to stay in the lungs.
These conditions are almost only found in asbestos worker and miners and not in the general public. In the case of high exposure to asbestos cigarette smoking is also a very large contributor to the cancer caused by asbestos.
Had the asbestos studies been allowed to be fully peer reviewed before legislation was passed we would probably still be using asbestos in many applications today with more protection for asbestos workers and minors and restrictions on some products such as asbestos powder and asbestos cloth. Unfortunately the legislators won't review their laws once they are passed and the western world is spending needless billions of dollars in asbestos abatement in every old building that is renovated.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
} From: "Chiphead" {chiphead-at-sbcglobal.net}
: : Warning: Non scientific answer: : : I remember an edition of "This Old House" or one of those types of shows : where they took some shingle samples to a lab for testing. : : The lab used polarized microscopy to do at least an "initial" screening : and determine that the shingles "most likely" contained asbestos. : : The "would do some additional testing" to verify (it was, and the house : needed to have a tent built around it, etc., etc.). I assume the : additional testing was something along the lines of TEM, etc. : : The conclusion I drew was that if you had materials that you were : expecting asbestos, this (polarized examination) was a good screening : tool. If the fibers didn't change from one specific color to another as : the analyzer was rotated, you didn't have asbestos. If they did, you : probably have asbestos, but there are other fibers that may do the same : thing. : : These were also fibers, not dust, and I don't know if that would affect : the results or not. : : Sorry if this wasn't useful. : : John R. : : -----Original Message----- : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] : Sent: Saturday, May 22, 2004 9:23 AM : To: microscopy-at-ns.microscopy.com : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical : Microscopy : : : : ------------------------------------------------------------------ ------ : ------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------ : ------- : : Below is the result of your feedback form (NJZFM-ultra-55). It was : submitted by (antuni-at-aol.com) from : http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, : May 21, 2004 at 16:19:26 : ------------------------------------------------------------------ ------ : --- : : Email: antuni-at-aol.com : Name: Anthony Ribaudo : : Organization: Consultant : : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical : Microscopy : : Question: Can one identify asbestos only utilizing optical microscopy : without the aid of electron diffraction/TEM ? : : : Anthony Ribaudo : : : : ------------------------------------------------------------------ ------ : --- : : : : :
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 12:48:22 2004
I have a student in my lab who wants to do immunohist on mouse skin. He is looking at RPA. His current protocol for RPA is on cells. He was able to get skin from mice treated with BPDE, he fixed the small strips of skin in 4% paraformaldehyde in Dulbecco's PBS. He fixed it for 20-24 hours, then put it in 70% ethanol followed by routine paraffin embedding. Was the fixation time too long? I always fixed for 4-6 hours tops. Will this affect his immunohist? Thanks for any help or advice on this.
Stacey Andringa
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 12:52:09 2004
I work in Cincinnati and we are experiencing our 17 year cicada invasion. I have never worked with insects and I am interested in fixing some for light microscopy. What fixative would work on them and how do I fix them - remove wings, pierce abdomen, etc.? This is for fun as much as education. Thanks for your help.
Stacey Andringa
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 12:53:49 2004
} For a FEG-HRTEM, what is the "line resolution" as } opposed to the "information resolution"? What is the } difference between each other? } } How could I achieve the "line resolution" and the } "information resolution" in TEM experiments? And, how } can I resolve (identify, or "see") the "line } resolution" and the "information resolution" } respectively from a state-of-the-art HREM image? } Dear Juha, The line resolution is the closest distance between two edges that can be resolved, and it can be pretty confusing. A better measure is point resolution, which can be determined from the first zero of the contrast transfer function at Shertzer focus. The information limit is the farthest extent of the Thon rings that can be achieved. To determine the point resolution, put a thin amorphous carbon specimen in the scope, adjust the height to eucentric height, make sure the scope is aligned as well as possible, go to a sufficiently high magnification so that the spatial frequency of the expected point resolution is larger than two--or better three--pixels (on a CCD, the actual elements, on film, the grain separation or resolution of a scanner), then take an image. Determine the power spectrum by taking the FFT of the image--easy on a CCD; on film, scan with a high-resolution scanner, then take the FFT. The spatial frequency of the first zero is the point resolution of the image. The better the scope is adjusted and the better the area of the specimen, the closer you will come to the true resolution of the scope. The information limit is best seen by shifting the image during the exposure. This produces Young's fringes in the power spectrum, which makes the extent of the Thon rings easier to see. The service engineers who set up our scopes used a switch attached to a shift coil to do this test. In order to achieve the best resolution and information transfer possible, make sure your scope is as well aligned as possible. If your specimen is too thick, or otherwise unsuitable, you may not achieve the specified performance, so don't blame the equipment in this case. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 13:59:59 2004
OK, you sucked me in to asbestos and so here goes...........
Almost all of the minerals in existence were identified and characterized by PLM at one time. Not all of these mineral were done as well as the rest but still PLM is a powerful tool in the hands of an experienced and trained microscopist. All of the legally defined asbestos can be identified by PLM. Dispersion staining is a very useful tool for this purpose. Vinyl tile, mortar, ceiling tiles, fluffy insulation, wall plaster, and black or gray or green mastic - sample prep remains the kingpin of all microscopical analysis just as it is in all analytical testing.
My understanding is the current alert limits were based on measurements made in asbestos mines and mills and extended to "normal" areas. This level may be too low and in my opinion is a result of political games. It seems some people have extreme sensitivity to asbestos and develop terminal conditions at very low exposure levels. It is also my understanding these studies were originally based on adults who were exposed as young adults and older. I have not seen studies bases on exposure of children to asbestos. So were do we draw the line? Yes, smoking makes it 400X more likely to develop health complications (at least that was the number quote when I did my asbestos certification training in Ohio. Not everyone who was exposed to high levels developed health complications. I had a boss who crawled through asbestos rewiring naval ships as summer work. He was a long term chain smoker, and he lived into his 80's.
Government regulations created a industry of abatement and identification. I have had the good fortune to participate at the identification and quantification level for a number of years. The thing to remember is it doesn't matter your personal beliefs or current scientific data when you're compiling with a law, it is what the law says it is.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gordon,
It is hard to believe other forms of asbestos: amosite asbestos(grunerite), actinolite-tremolite asbestos, anthophyllite (asbestos)and crocidolite asbestos(riebeckite) are not harmful. Have any studies been done on this? Chrysotile asbestos makes up to +90% of all asbestos in North America. (please correct if somebody has published figures on this).
Asbestos has been linked to mesothelioma and, also, asbestos is the cause of asbestosis(also harmful to humans). Isn't it?
I agree. If asbestos studies had been fully reviewed we would still using asbestos it has unique chemical and physical properties, especially for a mineral. The politicians used asbestos as a political football first scaring the public then passing legislation covering asbestos in schools, AHERA. Yes many problems have occurred as a result of this. The regulation should be reviewed and changed(if if hasn't already been).
Yes, asbestos can be identified by polarized light microscopy, not OLM or PCM. Using dispersion staining techniques to determine refractive indices, a properly prepared asbestos fiber can be identified, by PLM. Other measured properties: pleochroism, sign of elongation, and angle of extinction plus physical characteristic further aid the PLM analyst in the proper identification. Preparation of the fiber and proper training of the analyst are the critical factors here. Binder/matrix effects may hinder the analysis, for example chrysotile in vinyl floor tile.
Where are all the McCrone(ies) on this? Maybe this subject has been hashed out in the past. If so pardon me for the repeated redundancy.
Regards,
Timothy J. Bard, Scientist Scanning Electron Microscopy OSRAM SYLVANIA Towanda, PA
U
-----Original Message----- } From: Gordon Couger [mailto:gcouger-at-provalue.net] Sent: Saturday, May 22, 2004 5:09 PM To: Chiphead; 'by way of MicroscopyListserver'; microscopy-at-ns.microscopy.com
Only one of the forms of asbestos is harmful to humans. Chrysotile and then only if it is inhaled and the particle size is such it lodges in the lung so it can form a site for cancer to get a start. Chrysotile is only a small percentage of asbestos and the particle size must fall in the 5 to 15 micron range to stay in the lungs.
These conditions are almost only found in asbestos worker and miners and not in the general public. In the case of high exposure to asbestos cigarette smoking is also a very large contributor to the cancer caused by asbestos.
Had the asbestos studies been allowed to be fully peer reviewed before legislation was passed we would probably still be using asbestos in many applications today with more protection for asbestos workers and minors and restrictions on some products such as asbestos powder and asbestos cloth. Unfortunately the legislators won't review their laws once they are passed and the western world is spending needless billions of dollars in asbestos abatement in every old building that is renovated.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
} From: "Chiphead" {chiphead-at-sbcglobal.net}
: : Warning: Non scientific answer: : : I remember an edition of "This Old House" or one of those types of shows : where they took some shingle samples to a lab for testing. : : The lab used polarized microscopy to do at least an "initial" screening : and determine that the shingles "most likely" contained asbestos. : : The "would do some additional testing" to verify (it was, and the house : needed to have a tent built around it, etc., etc.). I assume the : additional testing was something along the lines of TEM, etc. : : The conclusion I drew was that if you had materials that you were : expecting asbestos, this (polarized examination) was a good screening : tool. If the fibers didn't change from one specific color to another as : the analyzer was rotated, you didn't have asbestos. If they did, you : probably have asbestos, but there are other fibers that may do the same : thing. : : These were also fibers, not dust, and I don't know if that would affect : the results or not. : : Sorry if this wasn't useful. : : John R. : : -----Original Message----- : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] : Sent: Saturday, May 22, 2004 9:23 AM : To: microscopy-at-ns.microscopy.com : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical : Microscopy : : : : ------------------------------------------------------------------ ------ : ------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------ : ------- : : Below is the result of your feedback form (NJZFM-ultra-55). It was : submitted by (antuni-at-aol.com) from : http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, : May 21, 2004 at 16:19:26 : ------------------------------------------------------------------ ------ : --- : : Email: antuni-at-aol.com : Name: Anthony Ribaudo : : Organization: Consultant : : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical : Microscopy : : Question: Can one identify asbestos only utilizing optical microscopy : without the aid of electron diffraction/TEM ? : : : Anthony Ribaudo : : : : ------------------------------------------------------------------ ------ : --- : : : : :
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 14:15:01 2004
There are three commonly employed definitions of "HRTEM resolution". All depend on how information is transferred by the objective lens from the specimen "exit-surface wave" to the image intensity spectrum (the Fourier transform of the image intensity).
(1) "Fringe" resolution, or "line" resolution (sometimes called "lattice-plane" resolution), is measured from the highest spatial frequency that is present in the image intensity spectrum (and is thus detectable in the optical diffractogram). It may include non-physical detail, since it may include components from a "second-order" or "non-linear" interference generating a half-spacing term with spatial frequency up to twice that of the highest-frequency diffracted beam passed by the objective aperture and the convergence cross-coefficient envelope ["Resolution-damping functions in non-linear images", M.A. O'Keefe, in 37th Ann. Proc. EMSA, San Antonio, Texas (1979) 556-557]. For a properly aligned microscope, it is not affected by microscope spread-of-focus and depends only on factors such as vibration or detector MTF.
(2) "Linear-image" resolution, or "information-limit" resolution, is measured by the highest spatial frequency transferred linearly from the amplitude spectrum (the specimen "exit-surface wave") to the image intensity spectrum with any old phase. Transferred frequencies fall within one or more passbands, but other (lower) frequencies may be blocked. Increased underfocus will push the convergence-limited damping function to higher frequencies, thus this limit (often called just the "information limit") to point resolution is a function of microscope spread-of-focus. Spread of focus is a function of objective lens chromatic aberration and variations in lens current and in electron beam energy (and of vertical vibration of the specimen within the lens).
(3) "Scherzer" resolution, or "structure-image" resolution (sometimes called "point" resolution or "point-to-point" resolution) is measured by the highest linearly-transferred spatial frequency that can be passed when no lower frequencies are blocked or passed with opposite phase. The Scherzer image is important because it is (an approximation to) a projection of the specimen structure (to a limited resolution) in the direction of the incident beam. For images obtained at the Scherzer "optimum" defocus, the Scherzer resolution is generally defined by the upper limit of the low-frequency same-phase passband. Scherzer resolution is a function of objective lens spherical aberration and electron wavelength.
FEG-HRTEMs can transfer spatial frequencies out to quite high values (their information limits are much better than their Scherzer resolutions). Since we know how linear transfer varies with objective lens defocus, we can use a series of images to produce a single reconstructed image containing correctly-phased components all the way out to the microscope information limit [see work by Van Dyck, Coene and Thust]. The latest issue of Microscopy Today shows an example of a reconstructed image with 0.78 Angstrom resolution, although the microscope used has a Scherzer resolution of only 1.7 Angstrom.
While the phases of linearly-transferred image components can be corrected so they can be used to extend resolution out to the microscope information limit, the non-lnear components that create the "fringe resolution" usually transfer no new information. These "cross-aperture" components can be frequency-doubled -- for example, a 200 beam from the specimen may interfere with a -200 beam to create a 400 image component. Then we will see spacings of a/4 in the image even though no information about the a/4 spacing was transferred from the specimen. If we managed to "dis-entangle" the image's non-linear 400 component with its a/4 spacing, it would still only provide us with information on a/2 spacings in the specimen.
So the short answer to your question is -- the difference between "line resolution" as opposed to the "information resolution" is that an image can contain specimen information out to the "information resolution", but any finer image spacings -- out to the "line resolution" -- are not new information about the specimen, but were generated by non-linear interferences in the objective lens.
Mike O'Keefe
Juha Dapper wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear microscopist experts, } } When I read literatures, I have been all the time } confused by the resolution definition in transmission } electron microscopy. Could you please kindly help? } } For a FEG-HRTEM, what is the "line resolution" as } opposed to the "information resolution"? What is the } difference between each other? } } How could I achieve the "line resolution" and the } "information resolution" in TEM experiments? And, how } can I resolve (identify, or "see") the "line } resolution" and the "information resolution" } respectively from a state-of-the-art HREM image? } } Thank you for your time. Wish you a nice weekend, } } -Juha } } } } } __________________________________ } Do you Yahoo!? } Yahoo! Domains – Claim yours for only $14.70/year } http://smallbusiness.promotions.yahoo.com/offer
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 15:36:21 2004
I have a water chiller from 1996 (Haskris RO75 with B H M + T options).
On checking the water level today, I noticed that the water was a bit green and hence needed to be changed. I drained the water storage tank, removed the nylon strainer, replaced it with a clean spare and as luck would have it I also needed to change the drain hose.
From experience I knew that when I first change the water, there is normally a rinsing out of the system and a noticeable amount of algae (? greenish gunk) comes out of the discharge pipe and I change the water another time before adding the anti-algecid and closing up the system.
Today I got some air into the system and when I turned it back on the water turned so turbid that I could not see the bottom coils in the tank. I repeated the draining/filling cycle at least 8 times until there was only a slight greenish cast to the water and I gave up until tomorrow.
Does anyone have any idea why this would happen now and not before? Is there any way that I can be assured that I have removed as much of the algae from the system as I can, other than to keep changing the water as long as I can see small clumps coming out into the water tank?
Thanks in advance, Pat Connelly Dept. of Biology Univ. of PA Philadelphia, PA 19104
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 15:48:37 2004
I seem to recollect that Neal Rowlands, who was Shared Experimental Facilities Manager in our Center some years ago, had done some asbestos research at McGill U. along the lines of your question. I recall him saying that the difference in lung damage caused by different forms of asbestos had to do with the difference in crystal shape. The tiny-needle shape was the nastiest.
Best regards,
Libby Shaw
} Subject: [Microscopy] RE: viaWWW: Identifying Asbestos Using optical } Microscopy } Date: Mon, 24 May 2004 12:26:01 -0400 } From: "Bard, Timothy J." {Timothy.Bard-at-SYLVANIA.com} } To: "Gordon Couger" {gcouger-at-provalue.net} , } "Microscopy (E-mail)" {microscopy-at-ns.microscopy.com} } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Address: MIT Room 13-4149 Tel: 617-253-5045 77 Massachusetts Avenue Email: elshaw-at-mit.edu Cambridge, MA 02139 Fax: 617-258-6478 http://web.mit.edu/cmse/www/ ****************************************************************
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 01:05:16 2004
To Students attending M&M2004 in Savannah, If you are a student that will be attending (or would like to the attend) the M&M meeting in Savannah this summer, we would appreciate your help by assisting at the sessions. You would be volunteering as a "projectionist" for various sessions, most likely ones you would attend anyway. There is a training session Sunday afternoon (details when you volunteer).
What are the benefits of volunteering? Volunteers will be given free registration--a badge, opening night reception ticket and a CD of the abstracts.
Who to contact: John Shields (see address below) at jshields-at-cb.uga.edu
-- John Shields E.M. Lab, 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 03:13:08 2004
I just want to get this clear and look at the consequences of what you write.
If a nonlinear component of frequency 2f is present in an image, will it have been damped by the ctf-envelope just like the corresponding linear component of frequency f? In that case the presence of a high-resolution component doesn't say much about the quality of a microscope or a micrograph as long as it is unclear whether the component is linear or frequency doubled. Now for the practical consequences: With what sort of specimens would you expect frequency doubling? Could it happen with protein crystals (10-20 nm thick) using a 200 or 300 kV microscope?
Philip
-----Original Message----- } From: Michael O'Keefe [mailto:MAOKeefe-at-lbl.gov]
(1) "Fringe" resolution, or "line" resolution (sometimes called "lattice-plane" resolution), is measured from the highest spatial frequency that is present in the image intensity spectrum (and is thus detectable in the optical diffractogram). It may include non-physical detail, since it may include components from a "second-order" or "non-linear" interference generating a half-spacing term with spatial frequency up to twice that of the highest-frequency diffracted beam passed by the objective aperture and the convergence cross-coefficient envelope
While the phases of linearly-transferred image components can be corrected so they can be used to extend resolution out to the microscope information limit, the non-linear components that create the "fringe resolution" usually transfer no new information. These "cross-aperture" components can be frequency-doubled -- for example, a 200 beam from the specimen may interfere with a -200 beam to create a 400 image component. Then we will see spacings of a/4 in the image even though no information about the a/4 spacing was transferred from the specimen. If we managed to "dis-entangle" the image's non-linear 400 component with its a/4 spacing, it would still only provide us with information on a/2 spacings in the specimen.
So the short answer to your question is -- the difference between "line resolution" as opposed to the "information resolution" is that an image can contain specimen information out to the "information resolution", but any finer image spacings -- out to the "line resolution" -- are not new information about the specimen, but were generated by non-linear interferences in the objective lens.
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 05:02:13 2004
} From experience I knew that when I first change the water, there is } normally a rinsing out } of the system and a noticeable amount of algae (? greenish gunk) comes } out of the discharge } pipe and I change the water another time before adding the } anti-algecid and closing up the system. } Dear Pat, Are you sure that the gunk is algae and not Cu oxide? If the latter, you should consider adding a corrosion inhibitor to the water. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 06:08:07 2004
I thought some people might be interested in a follow-up of this discussion. (Don't tell me if your not, just ignore me.)
I've taken some images of carbon film on our CM120 with a brand new LaB6-filament just to get an idea of the limitations of this microscope.
I get information limits between 15 and 5 Angstrom depending on magnification and defocus (only "biological" defocus values) and the information limit does not improve with decreasing magnification as suggested by CTF-explorer. In fact the opposite is true (at least for constant exposure levels). Some similar data for a FEG would be interesting.
One thing is clear: Reaching 10 Angstrom and slightly better should not be a problem with a well maintained LaB6 microscope.
For the details see the link "reference data for the CM120" at http://www.csb.ki.se/users/philip/philown.html. This is a big word file so you have to be patient when you download it.
Philip
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 06:31:39 2004
That's exactly what I was thinking, as well. Do you have any good recommendations for use as a corrosion inhibitor?
Thanks,
David
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Bill Tivol {tivol-at-caltech.edu} 05/24/2004 06:24 PM
To: microscopy-at-msa.microscopy.com cc: Subject: [Microscopy] [Detected by Millipore as possible Spam] Re: Haskris water chiller/recycler
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On May 24, 2004, at 1:48 PM, Pat Connelly wrote:
} From experience I knew that when I first change the water, there is } normally a rinsing out } of the system and a noticeable amount of algae (? greenish gunk) comes } out of the discharge } pipe and I change the water another time before adding the } anti-algecid and closing up the system. } Dear Pat, Are you sure that the gunk is algae and not Cu oxide? If the latter, you should consider adding a corrosion inhibitor to the water. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 07:53:51 2004
Bill I'm pretty sure I have Cu oxide along the walls of my Haskris chiller as it won't come off. What corrosion inhibitor do you suggest and how do I remove the Cu oxide already present?
Tony Greco
-- Anthony M. Greco Electron Microscope Manager College of Marine Science University of South Florida 140 7th Avenue S. St. Petersburg, Fl 33701
I'm glad that this issue is still being addressed. I have a Haskris chiller (since 1988) which services our Philips CM10 TEM. We continually have this "green algae" in the water and which seems to collect around the filter unit in the system. I have been concerned about this in the past, and asked our service engineer, but it doesn't appear to impede the flow of water or the operation of the chiller. (We have had other minor problems with the chiller that B & G has fixed, but nothing connected to the water). I was interested to hear about the possibility of it due to Cu oxide. I would also like to know what corrosion inhibitor can be added to the water.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: David_Bell-at-millipore.com [mailto:David_Bell-at-millipore.com] Sent: Tuesday, May 25, 2004 7:43 AM To: Bill Tivol Cc: microscopy-at-msa.microscopy.com
Hi Bill,
That's exactly what I was thinking, as well. Do you have any good recommendations for use as a corrosion inhibitor?
Thanks,
David
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Bill Tivol {tivol-at-caltech.edu} 05/24/2004 06:24 PM
To: microscopy-at-msa.microscopy.com cc: Subject: [Microscopy] [Detected by Millipore as possible Spam] Re: Haskris water chiller/recycler
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On May 24, 2004, at 1:48 PM, Pat Connelly wrote:
} From experience I knew that when I first change the water, there is } normally a rinsing out } of the system and a noticeable amount of algae (? greenish gunk) comes } out of the discharge } pipe and I change the water another time before adding the } anti-algecid and closing up the system. } Dear Pat, Are you sure that the gunk is algae and not Cu oxide? If the latter, you should consider adding a corrosion inhibitor to the water. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 08:56:31 2004
Thank you, Michael and Bill, for your kind reply. I really appreciate your remarkable answers to my questions.
After carefully reading your answers, I've got more questions.
1. If the "line resolution" is so confusing and untrustworthy, why TEM manufactures used to apply it to claim the quality of a microscope?
2. If an HREM iamge was taken without a certain objective aperture, how can we tell if a high-frequency spacing present in the image (or the corresponding spot in its FFT) is due to linear transfer or just by "cross-aperture" non-linear interference?
3. About the "non-linear interferences". Do they happen already during the process of electron-specimen interaction, or after the back-focal plane of the objective lens during the electron wave propagation?
4. In Michael's email, it is said "Since we know how linear transfer varies with objective lens defocus, we can use a series of images to produce a single reconstructed image containing correctly-phased components all the way out to the microscope information limit [see work by Van Dyck, Coene and Thust]."
Did you mean that the reconstruction method using a focal-series can only apply to the linear transfer case, i.e. the case of a weak phase object? If not, how the non-linear interference effects are treated?
5. In Bill's email, you mentioned that "The information limit is best seen by shifting the image during the exposure. This produces Young's fringes in the power spectrum,".
Could you please tell me in more details about how to "shift the image to produce Young's fringes"? This may be very practical and helpful to me.
Again thank you very much for your instructions.
-juha
__________________________________ Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger. http://messenger.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 10:42:49 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America On May 24, 2004, at 1:48 PM, Pat Connelly {psconnel-at-sas.upenn.edu} wrote: } } From experience I knew that when I first change the water, there is normally a rinsing out of the system and a noticeable amount of algae (? greenish gunk) comes out of the discharge pipe and I change the water another time before adding the anti-algecid and closing up the system. etc. } } } Dear Pat, } Are you sure that the gunk is algae and not Cu oxide? } If the latter, you should consider adding a corrosion } inhibitor to the water. } Bill Tivol, PhD } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 tivol-at-caltech.edu ======= Bill, I took a sample from the fourth change of water and examined it under a light microscope. I saw mostly tiny cellular clumps and cellular sheets with a few very small, redish, spike like, crystals embedded into them and that is why I thought of algae. I do agree that the green color most likely comes from the copper because there is no light in the system for photosynthesis. I have sprinkled dichlorophene onto the water each time that I have changed it since the first fill-up. The label states that it is both a fungicide and a bactericide. As the water supply, I use reverse osmosis water and some Philadelphia tap water so that there are some ions. The ph of both sources is usually acid, running about pH 5.5. I did not check the pH of the water that I drained yesterday but after rinning overnight the water resevoir is now also at pH 5.5.
I am interested in your response as to a corrosion inhibitor. Pat Connelly
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 12:24:55 2004
Rosemary; My recollection from the asbestos work I did at Lockheed was that most of the asbestos fibers could produce mesothelioma, but that in North America, chrysotile is the predominant type of asbestos. Sources of exposure vary, but the connection to mesothelioma was first noticed after WWII when a significant population that also smoked heavily was involved in refitting steam ships that had large amounts of asbestos insulation. The death of actor Steve McQueen highlighted the fact that even something like dirt bike riding in areas naturally high in asbestos could lead to a high risk exposure.
John Mardinly Intel
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-csiro.au] Sent: Sunday, May 23, 2004 4:28 PM To: microscopy-at-msa.microscopy.com
This is off the topic of identifying asbestos, but thought I should comment that just yesterday there was a program on the radio (streaming audio available at http://www.abc.net.au/rn/talks/bbing/, transcript available by Thursday) about new cases of asbestosis and mesothelioma showing up. The young people (20s and 30s) now getting this lived in high asbestos areas as kids, were children of builders (and builders themselves) who demolished or renovated fibro houses, etc. Perhaps just an Australian problem.... Rosemary
} From: "Gordon Couger" {gcouger-at-provalue.net} } Date: Sat, 22 May 2004 16:09:06 -0500 } To: "Chiphead" {chiphead-at-sbcglobal.net} , "'by way of MicroscopyListserver'" } {antuni-at-aol.com} , {microscopy-at-ns.microscopy.com} } Subject: [Microscopy] Re: RE: viaWWW: Identifying Asbestos Using ptical } Microscopy } } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------} - } } Only one of the forms of asbestos is harmful to humans. Chrysotile } and then only if it is inhaled and the particle size is such it } lodges in the lung so it can form a site for cancer to get a start. } Chrysotile is only a small percentage of asbestos and the particle } size must fall in the 5 to 15 micron range to stay in the lungs. } } These conditions are almost only found in asbestos worker and miners } and not in the general public. In the case of high exposure to } asbestos cigarette smoking is also a very large contributor to the } cancer caused by asbestos. } } Had the asbestos studies been allowed to be fully peer reviewed } before legislation was passed we would probably still be using } asbestos in many applications today with more protection for } asbestos workers and minors and restrictions on some products such } as asbestos powder and asbestos cloth. Unfortunately the legislators } won't review thier laws once they are passed and the western world } is spending needless billions of dollars in asbestos abatement in } every old building that is renovated. } } Gordon } Gordon Couger gcc-at-couger.com } } I collect links on information related to light microscopes. } http://www.couger.com/microscope/links/gclinks.html } Please forward any links or information you think might be useful to } others. } Microscope Manual at www.science-info.org } } } From: "Chiphead" {chiphead-at-sbcglobal.net} } } : } : Warning: Non scientific answer: } : } : I remember an edition of "This Old House" or one of those types of } shows } : where they took some shingle samples to a lab for testing. } : } : The lab used polarized microscopy to do at least an "initial" } screening } : and determine that the shingles "most likely" contained asbestos. } : } : The "would do some additional testing" to verify (it was, and the } house } : needed to have a tent built around it, etc., etc.). I assume the } : additional testing was something along the lines of TEM, etc. } : } : The conclusion I drew was that if you had materials that you were } : expecting asbestos, this (polarized examination) was a good } screening } : tool. If the fibers didn't change from one specific color to } another as } : the analyzer was rotated, you didn't have asbestos. If they did, } you } : probably have asbestos, but there are other fibers that may do the } same } : thing. } : } : These were also fibers, not dust, and I don't know if that would } affect } : the results or not. } : } : Sorry if this wasn't useful. } : } : John R. } : } : -----Original Message----- } : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] } : Sent: Saturday, May 22, 2004 9:23 AM } : To: microscopy-at-ns.microscopy.com } : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical } : Microscopy } : } : } : } : ------------------------------------------------------------------ } ------ } : ------ } : The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } : To Subscribe/Unsubscribe -- } : http://www.msa.microscopy.com/MicroscopyListserver } : On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } : ------------------------------------------------------------------ } ------ } : ------- } : } : Below is the result of your feedback form (NJZFM-ultra-55). It } was } : submitted by (antuni-at-aol.com) from } : http://microscopy.com/MicroscopyListserver/MLFormMail.html on } Friday, } : May 21, 2004 at 16:19:26 } : ------------------------------------------------------------------ } ------ } : --- } : } : Email: antuni-at-aol.com } : Name: Anthony Ribaudo } : } : Organization: Consultant } : } : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos } Using ptical } : Microscopy } : } : Question: Can one identify asbestos only utilizing optical } microscopy } : without the aid of electron diffraction/TEM ? } : } : } : Anthony Ribaudo } : } : } : } : ------------------------------------------------------------------ } ------ } : --- } : } : } : } : } : } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 12:56:12 2004
Does anyone else think it might be a useful exercise to try and establish expiration dates for chemicals used in microscopy labs? I understand that chemicals "expire" based upon what they are, what use they are intended for, how they are used, what level of accuracy/repeatability/etc. is expected from their use, how they are stored, and many other factors. I have also read Rande Kline's discussion of the problem in the July/August 2000 Microscopy Today on the difficulties of establishing such dates (recommended).
The problem is that some work requires lab certification and strict laboratory guidelines requiring that all chemicals be labeled with expiration dates----whether or not those dates have any meaningful or consistent basis in "reality". So far, I've been unable to find any such set of standards and my strong impression is that labs set up their own arbitrary chemical rotation regimen.
It might reasonably be suggested that MSA could be a clearinghouse for establishing such standards for microscopy-related research. If enough people think this is a useful project, I would be willing (please somebody, stop me!!) to volunteer to put together a draft proposal. As a starter, it would be useful to know how other labs handle this issue and why.
Any thoughts?
Thanks, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 13:36:42 2004
I've had a similar problem with my chiller system, though mine got clogged up pretty well before I figured out what was going on. The turbo pump in my SEM was the culprit, I think. The water jacket on the turbo is aluminum and there are lots of brass fittings in the Haskris and the heat sinks in the SEM. I got a bunch of stuff that looked like green kitty litter out of the system that proved to have high aluminum and copper levels in it. I think there is some sort of an electrochemical gradient set up that causes some sort of corrosion product to build up over time (I'm a biologist, not a chemist). Small amounts of it look like green sand in the bottom of the reservoir. I cleaned the bigger clogs out of the water jacket with alternating dilute HCl and NaOH using a big syringe and Tygon tubing, rinsing with DIW in between (Rube Goldberg lives on). It's been fine for a few months now, but I keep an eye on the flow rates.
Robert Simmons
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 14:18:48 2004
On May 25, 2004, at 4:43 AM, David_Bell-at-millipore.com wrote:
} That's exactly what I was thinking, as well. Do you have any good } recommendations for use as a corrosion inhibitor? } Dear David, When I was at Albany NY, we used a product called Aquatreet 42, a Mo-based inhibitor, and, judging from measurements of the water flow through the lenses each year, there was little corrosion build-up. That product can be obtained in a minimum of 5 gal--enough to last a lifetime--from Aqua Labs, Inc., P.O. Box 645, Amesbury MA 01913. I just spoke recently with Tom Cass, (800) 343-0213, and they still sell the product to people in your area. He also told me that Skasol makes a comparable product for the West Coast people. David Marchman, (800) 839-1000, is the contact for Skasol, and Chris Killian, (310) 749-8807 is the technical person. I have no affiliation with Aqua except as a satisfied customer, and none with Skasol except as a potentially satisfied customer. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 14:25:29 2004
Hi, Try dissolving it in 10% HCl. If it disappears its a Cu compound.
Best Regards,
Steve
Steve Buckingham Service Manager Kratos Analytical Inc., 100 Red Schoolhouse Road, Bldg A Chestnut Ridge, NY 10977 ph (845) 426 6700 ext 202 fx (845) 818 {mailto:4095steve-at-kratos.com} 4095
steve-at-kratos.com
-----Original Message----- } From: psconnel-at-sas.upenn.edu [mailto:psconnel-at-sas.upenn.edu] Sent: Tuesday, May 25, 2004 11:54 AM To: Bill Tivol Cc: Microscopy-at-msa.microscopy.com
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America On May 24, 2004, at 1:48 PM, Pat Connelly {psconnel-at-sas.upenn.edu} wrote: } } From experience I knew that when I first change the water, there is normally a rinsing out of the system and a noticeable amount of algae (? greenish gunk) comes out of the discharge pipe and I change the water another time before adding the anti-algecid and closing up the system. etc. } } } Dear Pat, } Are you sure that the gunk is algae and not Cu oxide? } If the latter, you should consider adding a corrosion } inhibitor to the water. } Bill Tivol, PhD } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 tivol-at-caltech.edu ======= Bill, I took a sample from the fourth change of water and examined it under a light microscope. I saw mostly tiny cellular clumps and cellular sheets with a few very small, redish, spike like, crystals embedded into them and that is why I thought of algae. I do agree that the green color most likely comes from the copper because there is no light in the system for photosynthesis. I have sprinkled dichlorophene onto the water each time that I have changed it since the first fill-up. The label states that it is both a fungicide and a bactericide. As the water supply, I use reverse osmosis water and some Philadelphia tap water so that there are some ions. The ph of both sources is usually acid, running about pH 5.5. I did not check the pH of the water that I drained yesterday but after rinning overnight the water resevoir is now also at pH 5.5.
I am interested in your response as to a corrosion inhibitor. Pat Connelly
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 14:33:40 2004
} I get information limits between 15 and 5 Angstrom depending on } magnification and defocus (only "biological" defocus values) and the } information limit does not improve with decreasing magnification as } suggested by CTF-explorer. In fact the opposite is true (at least for } constant exposure levels). Some similar data for a FEG would be } interesting. } Dear Philip, One of the acceptance tests for our Tecnai F30H was that we had to see 30 Thon rings at 2 um underfocus on a thin C specimen, which is slightly beyond the 0.34 nm graphite spacing. We redo this test periodically to be sure that the coherence of the beam is still up to par. In fact, the service engineers were able to get many more Thon rings than that. The spec on the info limit is 0.14 nm, but we can't get Thon rings that far out at 2 um underfocus. To get the 30 Thon rings, one has to find a good area of the C film, so we don't expect that good a performance for biological specimens. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 15:42:30 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (amcelwai-at-bucknell.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 25, 2004 at 14:55:05 ---------------------------------------------------------------------------
Email: amcelwai-at-bucknell.edu Name: Andrew
Organization: Bucknell University Biology Department
Title-Subject: [Microscopy] [Filtered] MListserver: New Text/Manual
Question: Hi all,
I am a graduate student at Bucknell University and I am currently studying amphibian physiology. My thesis project involves studying skeletal muscles and liver tissue using both transmission electron microscopy, and light microscopy. I am interested in learning about new techniques in microscopy, or histology and I was hoping that someone could recommend a book that covers curent methodologies and, or the best ways of doing histology/microscopy research. All of the literature I have is from the 70's and the 80's and some of my colleagues suggested to me that my sources of information are a tad bit outdated.
I have used nanoplast resin for the first time following instructions provided with the kit. When I came to examine the sections I found there had been poor infiltration. I would welcome any suggestions from people that have worked with the resin that could resolve this plus any other help tips. Do I need to do a resin infiltration series similar to Araldite. I did email Pelco (manufacturer of the resin) about this but so far not had a response.
thanks Therese
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 19:14:18 2004
I had an Ice Wagon and a Nesslab chiller system. The green color you see MIGHT BE algae. Everyone blames the green color on algae.
I know from chiller experience and being a chemist that copper pipes corrode first to a black thin film of copper oxide material on the walls of chiller copper pipes. Next a deposit of copper carbonate forms over the initial copper oxide coating and on PE cartridge filters. The black color only forms on copper surfaces.
Green deposits in chiller lines in my lab were always thicker AFTER going through a diffusion pump. We had whole house filters before the DPs and they collected a green scale. The lines between the filter and the DP inlet were relatively clear of green deposits. Seems backwards, doesn't it? FYI.
TESTING: Remove the scale and look under a microscope as you add dilute HCl. If it bubbles, it's most likely lime green copper carbonate, as I had. To find out if it was copper and carboante, I would use the regular green water to test for copper ions first. Take the green chiller solution and add HCl. It will dissolve the turbidity and form a nice clear green solution. Add enough ammonium hydroxide to make the solution fairly basic. If copper ions are present, the solution will turn deep blue. Algae do not do this. Next add more HCL and the solution will turn green again. If your solution does this, it's copper carbonate. If you would like to reconfirm the blue color again, you can repeat the NH4OH treatment on the same sample of water. This reversible color change is not possible with algae. IMHO.
To test for carbonate, add some HCl to a new sample to just barely get a clear solution. Adjust the pH to 7-9 with ammonium hydroxide. Don't use NaOH as it has carbonate in it. Add some clear barium chloride solution to this basic solution. Enough carbonate ions will remain dissolved to cause a white precipitate of BaCO3 to form. That probably confirms the gas was originally CO2. If you then add HCl, the white barium carbonate precipitate disappears to form carbonic acid and barium chloride. This chemical behavior confirms CO3 for sure.
If you find the problem is algae, then you can add a small amount of a quaternary ammonium salt. This extremely long named chemical will end with quaternary ammonium salt and can be bought at any swimming pool supply as an algicide. Estimate the volume of your system and add the recommended amount. As I recall, a quart treats about 5,000 to 10,000 gallons of pool water. So you will add a very small amount, like teaspoons or tablespoons of it. It will kill the algae, if that's really the problem. Any pool owner can help you that uses iso-cyanurates for hypochlorous acid (chlorine) disinfection. He might give you a few mls and save you the trip to the pool store for a 40 year chiller supply of QAS long chained surfactant.
You can state, "I have plastic pipes, so where is the copper coming from in my system?" Our chiller coils were made of copper tubing and the reservoir tank had copper tubing showing in our units. They're the source of the copper ions. In our system we also had copper supply lines going to various labs. A mistake! There are other copper sources too. Ask your serviceman about potential EM sources like embedded lens cooling lines of fittings. Air is normally the source of the CO2 because the tank is normally open to air.
} From experience, we learned the hard way to run distilled water. I then put a tablespoon of bicarbonate of soda in the water in case the chiller water tried to go acidic. This trace ion concentration cuts down on the corrosive nature of pure distilled water. This bicarb is not necessary, unless you know the pH in your system drops over time. "Isn't that the source of the carbonate?" No, we had the problem when we used chloramine-T and no bicarb. We used bicarb after we switched to distilled water for trace ions and to slow down any decrease in pH.
You can get into a green color spiral with chloramine-T (CT). You add CT and the water eventually turns green. You measure the available chlorine after awhile and it's too low. You add more CT and the available chlorine goes up. After some time the available chlorine drops and the g