I would like to do some tomography of vesicles in cryo conditions, can anyone recommend what grid type I should use and what sort of support film would be best? Also for negative stain tomography which grids and support films should be best?
Many thanks, Anna Young
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 1 09:35:06 2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 1 13:08:54 2004
Thank you all for the great suggestions both on and off line. XRF, etc, however, are not available. The soil testing is being conducted with already well established protocols. The hope was/is to use the SEM/EDAX to look at P associations in the soil--perhaps associated with clay, Al, Fe, or Ca. Using mapping we have been able to locate some P-rich regions. We have tried RGB mixing to find regions where there are associations(P with other elements).
I hope this discussion has helped others as much as it has helped me. Hopefully, I can contact any of you offline when I get stuck with another 'where do I go from here question'.
Regards, Winnie
Edwina W. Westbrook Electron Microscopy Laboratories Agricultural Research Station M. T. Carter Building, room 129 P.O.Box 9061 Virginia State University Petersburg, VA 23806 (804)-524-5659 fax (804)-524-5622 ewestbro-at-vsu.edu
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If I understand your problem, what you (or your colleagues) are tying to determine is what mineral phase the P is present in. This would require that you analyze individual particles, and enough particles enriched with P to say something about the sample. If you are making maps of your sample you must be interested in some spatial features of P distribution in your sample. While X-ray mapping is a way to get at this, you might want to try a different prep technique to improve the mapping results. I recall reading an article on preparing polished cross sections of soil samples in the July 2004 Microscopy and Analysis that dealt with archaeological soil samples and might be of some use.
Hope this helps,
Bryan Bandli
Edwina Westbrook wrote:
} I should have been a bit more detailed. Remember, I am not the soil scientist, however, wet chemistry is the method of testing...spectroscopy (ICP) and chromatography. Those methods are performed in the soil lab. My task is simply to place a dbl sticky SEM stub into a soil sample, coat with carbon and view in the SEM (randomly selected areas), generate a spectra and maps. SEM/EDAX is in addition to the well documented soil analysis methods. } } Wanted to get some feedback or direction from all of you who are far more knowledgeable and generate discussion that may benefit others as well. } } Thank you. } } Winnie } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 1 15:59:02 2004
} I would like to do some tomography of vesicles in cryo conditions, can } anyone recommend what grid type I should use and what sort of support } film would be best? Also for negative stain tomography which grids and } support films should be best? } Dear Anna, We use Quantifoils, and they work very well for cryotomography. Get 200-mesh grids, since smaller mesh sizes will occlude the specimen at high tilt angles. I'd use carbon film for negative stain tomography. Carbon-formvar will also work well with essentially no loss of resolution, and the film is stronger under some circumstances than just carbon, but, since the formvar is not conducting, sometimes a carbon-formvar film will be less stable than just carbon. Again, use 200-mesh or larger grids. I am not affiliated with Quantifoil or other grid suppliers (coated or not), except as a 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 Wed Dec 1 16:02:57 2004
Want to determine the PSF of our Confocal Scanning Laser Ophthalmoscope (used to image the retina in the eye, but very similar in design to the Confocal Microscope). Unfortunately, I only had access to 10µm beads and they have a 12px diameter when imaged at the highest magnification. Can I still derive the PSF from these images, or do I have to buy smaller beads? If yes, how should I go about it?
} Re: Using a Kodak MDS100 on a B&L Research I metallograph
Aha ! You went to the Amenex website ...
} We have an ETEC Autoscope SEM from 1969. We have problems with } the image when we rotate from 180 to 90 degrees. The images seem } like a compact shape (see the photos below). Does someone has } information that help us to fix this? } Not good image (seems like compact image) Oh, oh. Aspect ratio isn't adjusted correctly } This the correct form of the image Looks OK. } This is our ETEC Autoscope SEM: Wow. Very clean & neat setup.
Ken Converse may chime in on this, but my first impression is that your 'scope has an adjustment for specimen tilt that restores the apparent shape of the specimen so that a square looks square even when the specimen tilt would introduce foreshortening. You had it adjusted OK at zero degrees, and now it's exactly wrong at 90 degrees. It's been four years since I last looked at our ETEC Autoprobe (now hopefully ensconced at or near UC Berkeley) but memory says that this adjustment is associated with the magnification selector. There's a set of thumbwheels at the lower end, I think.
Be sure to watch those red rockets - the Tantalum electrolytics that were _our_ Autoprobe's weak links. I'd look for the ones that had turned black and replace 'em with foil electrolytics from our local Radio Shack. Used the same or higher voltage and the same microfarad rating. The cans were bigger but they worked fine.
I just finished a report in which I used the MDS100 with the Research I metallograph to make quite a few photomicrographs, using a proper green interference filter (Kodak standard issue) which allowed me to see the images in the microscope eyepieces much more easily than with the blue interference filter. This green filter blocks the longer wavelengths far more effectively than the plain green filter that I first tried. Now I get green images with the MDS100 ! And then I convert them to greyscale, of course.
Best regards to all, George Langford, Sc.D. amenex-at-amenex.com http://www.amenex.com/
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 1 23:53:12 2004
Anna Young wrote: ============================================================================ ============ I would like to do some tomography of vesicles in cryo conditions, can anyone recommend what grid type I should use and what sort of support film would be best? Also for negative stain tomography which grids and support films should be best? ============================================================================ ============ The Quantifoil® grids were developed for just this application. See URL http://www.2spi.com/catalog/grids/quantifoil-carbon-films.html
Another, but not as well proven possibility, for this application, would be our perforated silicon nitride membrane window grids, see URL http://www.2spi.com/catalog/instruments/perforated-membrane-window-grids. shtml
Disclaimer: SPI Supplies offers both types of grids so we would have a vested interest in seeing more of either of them being used.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 07:20:22 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lan.xu-at-fmi.ch) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, December 2, 2004 at 02:36:08 ---------------------------------------------------------------------------
Email: lan.xu-at-fmi.ch Name: Lan Xu
Organization: FMI
Title-Subject: [Microscopy] [Filtered] EM
Question: Dear sir or madam, I would like to know a potocol for Formvar-coat single slot grides. I need to do a consecutive serial section for EM stduy in brain sections, but I don't know how to coat the single slot grides, (I have the normal potocol for coat Mesh square grides) If you could give me a potocol for Formvar-coat single slot grides, I would be very gladful. Thank you in advence, Lan Xu
The following question was forwarded to me, so I'm passing it on to the collective for possible suggestions.
"Do you know of any ways of labeling/marking or otherwise making Cd in brain tissue visisble, either by light or TEM?
We are doing blood and brain studies on cadmium exposure in rats.
I have been doing the Cd analysis in the tissues (blood and whole brains), but we want to look at areas of the brain for Cd levels. That puts the sample amount in the 15mg range and for me to analyze, that gives me values in the ppt range, which is too low. So we'd have to pool brain samples to get enough.
I wondered if there was some way to label the Cd in tissue prior or after making slides with some countable/fluorescent or otherwise quantifiable materials. So he could do brain sections and figure it out."
This looks like way too little for EDS analysis and I know nothing about labeling techniques for low concentrations of heavy metals. 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.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 14:39:32 2004
Please note that the deadline for MSA Award nominations if fast approaching (December 15, 2004). Information is available in the current issue of Microscopy and Microanalysis (vol 10 (6): facing page 822).
Categories include 2 Distinguished Scientist Awards (Biological and Physical Sciences), the Burton Medal (young investigator), 2 Outstanding Technologist Awards, and the Morton D. Maser Distinguished Service Award.
Please feel free to contact me for additional information if needed.
William T. Gunning, Ph.D. Professor of Biochemistry and Cancer Biology; Pathology Medical College of Ohio Departments of Biochemistry and Cancer Biology; Pathology Medical College of Ohio BHSB 140 3035 Arlington Avenue Toledo, Ohio 43614-5806 Phone: 419-383-4131 or 3752 Fax: 419-383-6228 email: wgunning-at-mco.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 15:56:14 2004
Dear Randy, I recall an aluminum x-ray map of an Alzheimer's patient's dried brain tissue, showing Al in ppm levels. It was done in an automated EPMA (WDX) and took about 16 hours to collect, with a long dwell time on each point. The presenter said that the EPMA wasn't being used overnight, so why not. The dried tissue will concentrate the metal somewhat. The other possibility is to use the backscattered detector. This will only work if the Cd is concentrated somewhat in some areas, but it should show up bright against the organic background. Your levels may be too low for either of these techniques. The only other one I know that is more sensitive is an x-ray microscope. 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 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu} To: {microscopy-at-microscopy.com} Sent: Thursday, December 02, 2004 12:09 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (markjaco-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, December 2, 2004 at 11:23:50 ---------------------------------------------------------------------------
Email: markjaco-at-hotmail.com Name: Mark Jacobs
Organization: Minneapolis, washburn high school
Education: 9-12th Grade High School
Location: Minneapolis, MN USA
Question: What education level do you need to work in microscopy? Where are the programs?
I love biology and working with microscopes. thanks Mark
You probably already saw this but check out this website
www.scripps.edu/news/press/112304.html
wherein they announce a "ligand" that will detect heavy metals such as Hg and Cd but is not reactive with Fe, etc.
The article, "A Precipitator for the Detection of Thiophilic Metals in Aqua," is authored by Tobin J. Dickerson, Neal N. Reed, James J. La Clair, and Kim D. Janda and will appear in an upcoming issue of the Journal of the American Chemical Society.
It may be applicable in microscopy.
John -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 23:04:44 2004
How small are the vesicles you are interested in imaging? Next week at Cell Bio, NT-MDT is going to announce a new integrated product which puts an AFM into a microtome. The AFM will image directly from the block face, eliminating typical errors from slippage, pull-out, tears, or wrinkles, then do a 3D reconstruction from the resulting image stack. I think it might be a faster, more effective solution to your application. Anton Efimov, the inventor of this technique, will also be available for discussion at the show. NT-MDT is being distributed here in the US through Nanotech America. They will be in booth 1129-1130, if you are interested.
Caveat: MME is providing support for this project.
I hope this is helpful, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
At 09:39 AM 12/1/2004, Anna Young wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 23:17:11 2004
If you love biology, then I would recommend that you major in that field, but find a subspecialty that takes you into microscopy and imaging. Today there is a lot of work done in live cell imaging and in fluorescence.
I am sure that your guidance counselor can help you to find a good bio program. There are so many choices, from genetics to marine biology,etc. Be prepared for the long haul, however, because you will most likely need to get a PhD and then do post doctoral work.
There are also interesting jobs opening up in industry (biotechnology, nanobiology, etc.).
Good hunting!
Best regards Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
At 05:17 PM 12/2/2004, markjaco-at-hotmail.com wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 23:59:33 2004
Hi Mark, I'm happy to hear you're interested in Microscopy. I'm an instructor at Madison Area Technical College in Madison, Wisconsin. We offer a two year Associate in Applied Science Degree in Electron Microscopy. It covers both biological and materials. There is also Delta College in California that offers a two year EM Program. There are a lot of other 4 year colleges and graduate programs that offer many great opportunities in this field. Check out some websites, and if you have any questions, please feel free to contact me. Bill
Bill Carmichael wcarmichael-at-matcmadison.edu http://matcmadison.edu/electronmicros
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:markjaco-at-hotmail.com] Sent: Thursday, December 02, 2004 5:18 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (markjaco-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, December 2, 2004 at 11:23:50 ------------------------------------------------------------------------ ---
Email: markjaco-at-hotmail.com Name: Mark Jacobs
Organization: Minneapolis, washburn high school
Education: 9-12th Grade High School
Location: Minneapolis, MN USA
Question: What education level do you need to work in microscopy? Where are the programs?
I love biology and working with microscopes. thanks Mark
For a long time, we have been trying to operate the integrated SX-50 unit (CAMECA). Although the system comprise of only one WDS, we believe that is fine too. When we try to calibrate the spectro we could not be able to catch any peaks in the "expected" region, for example:
Fe Ka peak in LIF crystal should be around 48084 (sine-theta) but when we scan the pure iron standart we see the peak around 74064 (sine-theta)
Or;
Si Ka peak in PET crystal should be around 81444 (sine-theta) but when we scan the pure Si standart we see the peak around 106497 (sine-theta).
There is an evident peak shift, nearly 25000 (sine-theta).
I have noticed that the spectrometer limits exceed the suggested values. In the troubleshooting section of reference guide; it says that limits of the spectrometer, namely; pmin should be { 22000 and pmax should be } 83000. Also the difference (pmax-pmin) should be greater than 61000. Our pmin is 46616 and pmax is 108545. We have the mentioned difference (108545-46616=61929) but the limits are shifted evidently. It should be expected that if we lower these two values by 25000, these would be in the suggested limits and there would be no peak shift!
I have done the following steps to lower pmin and pmax:
1) I have tried to initialize the WDS by "INIT WDS" or even by "INIT" commands in the SU Local console.
2) I have tried to reset the WDS Motorola 68000 microprocessor by its switches
3) I have even tried to look up the machine codes in the debugging mode
4) I used all variations (as far as I understand) of "FIX" commands
5) and lastly I decided to change the pmin and pmax variables by "DEFI" command, but I could not, as they are "system protected variables"
I decided to choose another way for it; I calibrated Fe Ka line on pure Fe sample with LIF crystal. But as expected, the "peaks" were nearly at the same intensities with background noise as a result of peak shift.
I prepared a declaration file, defining a chemical shift of 25000. I knew it was very unpractical and somehow random. Such a chemical shift value causes the software crash a lot and keeps giving the error message ".... beyond spectrometer limits"
I am desperately seeking help, I assume it is not a great technical problem but I must confess that I am stuck. Any reply would be highly and sincerely appreciated.
Orkun ERSOY Hacettepe University Department of Geological Engineering Beytepe-Ankara / TURKEY 06532 Ph: +90 312 2977700 / 126
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 07:03:13 2004
The education level you need to work in microscopy you already have. It is the second word in your message. Formal schooling helps, of course, if you want to get paid for it. But don't let the schooling get in the way of your education. Even if you become a professional, always strive to be an amateur (from 'amator', 'amare') that is, one who does something out of love, rather than for financial gain.
Paul
-------------------------------------------
Work like you don't need the money; Love like you've never been hurt; Dance like nobody's watching. - Satchel Paige
(The opinions expressed here do not necessarily represent those of my employer or Purdue University)
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:markjaco-at-hotmail.com] Sent: Thursday, December 02, 2004 6:18 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (markjaco-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, December 2, 2004 at 11:23:50 ---------------------------------------------------------------------------
Email: markjaco-at-hotmail.com Name: Mark Jacobs
Organization: Minneapolis, washburn high school
Education: 9-12th Grade High School
Location: Minneapolis, MN USA
Question: What education level do you need to work in microscopy? Where are the programs?
I love biology and working with microscopes. thanks Mark
We could not find the offsett value established by the installation engineer which Michael Shaffer told, but instead of 150 000 we wrote 100 000 and followed the procedure which Graham Hutchinson told.
Now it works!
Everything seems fine now! Thanks all....
Best regards
Orkun ERSOY Hacettepe University Department of Geological Engineering Beytepe-Ankara / TURKEY 06532 Ph: +90 312 2977700 / 126
My problem was;
For a long time, we have been trying to operate the integrated SX-50 unit (CAMECA). Although the system comprise of only one WDS, we believe that is fine too. When we try to calibrate the spectro we could not be able to catch any peaks in the "expected" region, for example:
Fe Ka peak in LIF crystal should be around 48084 (sine-theta) but when we scan the pure iron standart we see the peak around 74064 (sine-theta)
Or;
Si Ka peak in PET crystal should be around 81444 (sine-theta) but when we scan the pure Si standart we see the peak around 106497 (sine-theta).
There is an evident peak shift, nearly 25000 (sine-theta).
I have noticed that the spectrometer limits exceed the suggested values. In the troubleshooting section of reference guide; it says that limits of the spectrometer, namely; pmin should be { 22000 and pmax should be } 83000. Also the difference (pmax-pmin) should be greater than 61000. Our pmin is 46616 and pmax is 108545. We have the mentioned difference (108545-46616=61929) but the limits are shifted evidently. It should be expected that if we lower these two values by 25000, these would be in the suggested limits and there would be no peak shift!
I have done the following steps to lower pmin and pmax:
1) I have tried to initialize the WDS by "INIT WDS" or even by "INIT" commands in the SU Local console.
2) I have tried to reset the WDS Motorola 68000 microprocessor by its switches
3) I have even tried to look up the machine codes in the debugging mode
4) I used all variations (as far as I understand) of "FIX" commands
5) and lastly I decided to change the pmin and pmax variables by "DEFI" command, but I could not, as they are "system protected variables"
I decided to choose another way for it; I calibrated Fe Ka line on pure Fe sample with LIF crystal. But as expected, the "peaks" were nearly at the same intensities with background noise as a result of peak shift.
I prepared a declaration file, defining a chemical shift of 25000. I knew it was very unpractical and somehow random. Such a chemical shift value causes the software crash a lot and keeps giving the error message ".... beyond spectrometer limits"
I am desperately seeking help, I assume it is not a great technical problem but I must confess that I am stuck. Any reply would be highly and sincerely appreciated.
Orkun ERSOY Hacettepe University Department of Geological Engineering Beytepe-Ankara / TURKEY 06532 Ph: +90 312 2977700 / 126
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 08:49:50 2004
Thanks for this suggested link. The article is available on-line now. If your institution has a subscription to the Journal of the American Chemical Society for on-line content, this link should work:
At 06:36 PM 12/2/2004, you wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
.................................................................... Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy Research Specialist, Principal University of Arizona (office: AHSC 4212A) P.O. Box 245044 (voice: 520-626-2824) Tucson, AZ 85724-5044 USA (FAX: 520-626-2097) (email: Cromey-at-Arizona.edu) .................................................................... http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 11:15:44 2004
I need suggestions on how to do micro-structure analysis on automotive emission catalytic converter (honeycomb). Can anyone tell me how to prepare SEM and TEM samples from the honeycomb? Does the process follow the basic procedure: cutting, grinding, polishing and ion-milling? Basically, I want to get noble metal particles' (such as Pt) size and dispersion on the supports, such as alumina.
Thanks in advance for your advice. Any suggestion would be highly appreciated!
******************************************* Juan Cai, Ph. D. Scientist Nanostellar 3603 Haven Avenue, Suite A Menlo Park, CA 94025 Phone: 650-4502267 (cell) Fax: 650-3681101 jcai-at-nanostellar.com *********************************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 11:23:03 2004
Regarding Al and Alzheimer disease, this was done much more convincingly using a nuclear microprobe - particle induced X-ray emission technique (PIXE) with matrix corrections using proton backscattering. See publications by Frank Watt and Geoff Grime + medical collaborators. Regarding Cd, PIXE would be much better than EDS. Firstly, you should not use Cd-L lines because of overlaps with potassium K-lines and this is really a big problem. Therefore EDS is in practice not possible to use, and PIXE can be used on K lines of Cd. A microprobe based on synchrotron radiation would be much better indeed (you may call it "x-ray microscopy"?). I suggest searching papers related to this method (SXRF).
Best regards
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz Materials Research Group iThemba LABS PO Box 722 Somerset West 7129 South Africa E-mail: przybylowicz-at-tlabs.ac.za Fax: +27-21-8433543 Phone: +27-21-8431166 (direct); 8431000 (reception) Cell: +27-82-563 7925 http://www.tlabs.ac.za xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
----- Original Message ----- } From: "Mary Mager" {mager-at-interchange.ubc.ca} To: "Tindall, Randy D." {TindallR-at-missouri.edu} Cc: "Microscopy" {microscopy-at-MSA.microscopy.com} Sent: Friday, December 03, 2004 12:20 AM
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Juan Cai wrote: ============================================================================ = I need suggestions on how to do micro-structure analysis on automotive emission catalytic converter (honeycomb). Can anyone tell me how to prepare SEM and TEM samples from the honeycomb? Does the process follow the basic procedure: cutting, grinding, polishing and ion-milling? Basically, I want to get noble metal particles' (such as Pt) size and dispersion on the supports, such as alumina. ============================================================================ = These systems consist of what is described as a) the honeycomb structure (which generally consists of low surface area alumina and is generally not of interest), b) the "wash coating" of high surface area alumina, usually gamma, on the internal surfaces of the honeycomb and c) the metal catalyst particles, generally platinum or platinum alloys, dispersed in the coated layer of high surface area alumina. These are true nanoparticles since they are rarely larger than 2 nm and usually less. What one generally wants to see is the degree of dispersion of the nano size metal particles in the high surface area alumina. We have generally found in our own laboratory, when we have tried to do these kinds of samples, the "basic procedures" as outline above generally fail. And the only way that we have ever found that works reliably is diamond knife thin sectioning on an ultramicrotome.
But this also means that you have to embed a piece of the honey comb substrate (but only a slight trace of the substrate structure remaining) that contains the alumina wash coat, keeping track of where you are on the sample, doing the diamond knife ultramicrotomy, and then examining the sections by TEM. You can not only image the size and degree of dispersion of the metal particles but you can also see areas of "hot spotting" which can have a dramatic effect on the efficiency of the catalyst. Some poisons such as lead can be imaged this way as well. One note of caution: I have encountered some workers who literally scrape off some of the high surface alumina and embed just the fines, an obviously much easier task, but you lose the all important spatial information when you do it this way. Others just look at the fines but it is not a very representative sample since the data comes only from those particles that are small enough to be electron transparent.
We have found that our own SPI-Pon 812 embedding resin works quite fine for this application but we would expect that at least some of the "Epon substitutes" offered by our competitors would work just as well. Cutting samples of this type is literally "hard" on anyone's diamond knife. We use the standard 45 degree knives which results in minimum compression. But be sure you don't waste money on an expensive "life science" knife for this kind of work since a 45 deg. "materials science" diamond knife will work just fine. See URL http://www.2spi.com/catalog/knives/materials.shtml for more information.
Disclaimer: Our firm does this kind of diamond knife thin sectioning as a laboratory analytical service for others. We are also suppliers of the recommended embedding resin and the materials science diamond knives.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 13:40:12 2004
We intend to surplus our Philips EM300 that is currently working well and is under service contract. We are looking for offers to remove and accept the unit, to make room for a new TEM.
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ William S. Marshall, Ph.D. ° ° ° Professor, Biology Department ° ° St. Francis Xavier University {°)\\\} { Box 5000 Antigonish NS B2G 2W5 Canada Phone 902-867-2482 Fax 902-867-2389 *************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 16:07:12 2004
Many thanks for all the suggestions and references! I have passed everything along to the person needing the information (and learned quite a bit myself).
Happy Holidays, 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.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 16:46:35 2004
A campus researcher has asked advice about obtaining a new SEM for our lab.
It's been a long time since anyone asked me about this, and I have let my knowledge of current instruments slide. Besides, it seems like they change so fast that it would be hard to keep up unless you are ready to get something new.
I would like to get advice about where to start looking, not which vendors, I know most of them, but where in the product/feature continuum we should start.
We have a 1986 vintage ISI WB-6 conventional SEM in the lab now. For the most part, it does OK. We don't need a large sample chamber or lots of whistles and bells. Our applications are varied as a central campus lab, but most are at least semi-biological, no hard core materials types.
The specific application of the inquiring researcher is looking at small pore patterns in diatoms to confirm species identifications.
I am especially interested in hearing about some of the new developments in SEM's since we got our ISI machine. Things like 'environmental' SEM, FEG developments etc. Also any updates on coating techniques beyond our conventional sputter coater would be useful.
Real world experiences would be great. You know, things like, this sounds good, but doesn't really add anything or this is a feature I never thought would be worth anything but I couldn't live without it now, type remarks.
With your help, I will be in a much better position to get the right instrument for my campus guy and be way better off when talking to vendors.
Thanks
Jon
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 17:24:37 2004
I feel that my short reply to Andrey’ question was slightly misunderstood… Let go with the points you raise and your comments on the spatial resolution of backscattered electrons (BSE) images:
A) “The size of the region from which backscattered electrons come is much bigger than the resolution in the image”: If you are talking of the spatial resolution of a backscattered electron image, I agree partially with this sentence. In my reply to Andrey, I should have written that the so-called “Surface Radius of BE” histogram generated by CASINO gives a CONSERVATIVE UPPER LIMIT of the spatial resolution of BSE images.
B) “The resolution in a secondary electron image corresponds to a distance much smaller than the size of the area from which the secondary electrons are emitted”: I fully agree, except that… I never spoke of secondary electron image in my reply to Andrey!
C) “If instead the graph is made to plot number of electrons against position (divide the first plot by 2pi times the radius) then the situation looks quite different”: In fact, you are suggesting to define a parameter J which is proportional to the true local current density in the region emitting the BSE. In such a case, one have to divide the number dN of BSE emitted by a surface element dS. Due to the axial symmetry of the problem, dS = (pi^2)[R(i+1) – Ri] = (pi^2)(delta R), where (delta R) is the increment of the radial distance Ri. Note that the term (pi^2) which is different from your 2pi…
With the raw data generated by the free software CASINO (http://www.gel.usherb.ca/casino/) and imported in a spreadsheet like Excel, I did this exercise and I have plotted three types of curve:
-- J (= dN/dS) –vs– distance R -- Cumulative J –vs– R -- Cumulative dN –vs– R
The “Cumulative J –vs– R” and “Cumulative dN –vs– R” curves are very similar to the normalised curves presented in figure 3.24 of Goldstein’s book (2nd edition). If anybody is interested by these results, please contact me off line and I will send you the figures.
Now the fundamental question is: “How to define the spatial resolution from these figures”. I agree that J (=dN/dS) is “sharply peaked at the center”, however one cannot ignore the other BSE coming from the rim around this peak. One have to make some assumption on the percent of BSE which significantly contribute to the signal coming from the emitting region and which are received by a BSE detector such as the solid-state detector. Conservatively, lets take a criterion of 60%. Andy, try this simple exercise on figure 3.24 of Goldstein’s book with the values of the Kanaya-Okayama radius given in table 3.2 of the same book. The results are as follows:
Energy of the primary electrons = 20 keV Criterion for defining the “spatial resolution” = 60% of the total number of BSE “Spatial resolution” for Copper (Z = 29) = 345 nm (nanometer) “Spatial resolution” for Aluminium (Z = 13) = 1350 nm (nanometer)
In my opinion and as daily demonstrated with any “classic” SEM, such figures clearly show that the “spatial resolution” of a BSE image will never match that of a SE image, except if one has the chance to work with special devices like the energy filtering system developed by Wells and commented in section 4.6.1 of Goldstein’s book.
-----Message d'origine----- De : Alwyn Eades [mailto:jae5-at-lehigh.edu] Envoyé : 30 novembre, 2004 08:29 À : Microscopy-at-MSA.Microscopy.Com Objet : Monte Carlo simulation of backscattered electrons
I think that the recent post on this subject from Jean-Paul BaÔlon is misleading - sorry Jean-Paul. I started to look into this but have not had time to finish the investigation but...
The size of the region from which backscattered electrons come is much bigger than the resolution in the image.
The resolution in a secondary electron image corresponds to a distance much smaller than the size of the area from which the secondary electrons are emitted. This is well known. See for example, page 197 of the latest edition of the Goldstein et al book.
Something similar happens with backscattered electrons, though this is more controversial. The plot that Jean-Paul refers to is a plot of number of backscattered electrons against distance. If instead the graph is made to plot number of electrons against position (divide the first plot by 2pi times the radius) then the situation looks quite different.
The number of backscattered electrons per unit area from the surface is sharply peaked at the center and details in the image can be seen at lengths related to this sharpness rather than the size of the whole area from which electrons come.
This is the explanation for the fact that backscattered images show details much smaller than a simple Monte Carlo result would suggest.
Alwyn -- ......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 4 02:54:06 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm not an expert on this ... but I think you'll find the structure is mechanically quite fragile. To get decent samples, you'll need to support the structure during preparation.
I'd first cut into smallish (~2 cm across and perhaps ~1 cm thick) and then embed in resin - I'd have thought a standard epoxy would be OK. Having done that, you can then proceed with standard metallurgical preparation techniques. I think you can get the information you need by SEM, so grinding and polishing is probably all you need to do.
Keep in mind, commercial auto catalysts contain a wide range of Pt group metals. If you want to distinguish the different elements, you will almost certainly need to use WDX, since EDX won't have sufficient energy resolution. -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 4 05:03:01 2004
Y. Sumi, Masanori T. Itoh, Takeshi Muraki, Takuro Suzuki (1996). Histochemical staining of cadmium with 2-(8-quinolylazo)-4,5-diphenylimidazole. Histochemistry and Cell Biology, Volume 106, Number 2: 223 - 227.
Regards,
Vera Santos Dental Medical School Institute of Biomedical Technology (ITB) University of Lisbon Portugal
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 4 07:19:02 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (somayyeh_kheiri-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, December 4, 2004 at 05:39:46 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] karyotype of verbascum
Question: Hello Dear all
I am doing meosic karyotype of Verbascum species on buds preseved in carnoy solution. but unfortunately the chromosomes are so little and I have problem counting them specially in the magnification of 100 LM. I would appreciate any helps and suggestion.
Have you tried FIB? In Situ liftout FIB should be able to make a satisfactory specimen of this type of material.
John Mardinly Intel
-----Original Message----- } From: Juan Cai [mailto:jcai-at-nanostellar.com] Sent: Friday, December 03, 2004 9:41 AM To: Microscopy-at-MSA.Microscopy.Com
Hi,
I need suggestions on how to do micro-structure analysis on automotive emission catalytic converter (honeycomb). Can anyone tell me how to prepare SEM and TEM samples from the honeycomb? Does the process follow the basic procedure: cutting, grinding, polishing and ion-milling? Basically, I want to get noble metal particles' (such as Pt) size and dispersion on the supports, such as alumina.
Thanks in advance for your advice. Any suggestion would be highly appreciated!
******************************************* Juan Cai, Ph. D. Scientist Nanostellar 3603 Haven Avenue, Suite A Menlo Park, CA 94025 Phone: 650-4502267 (cell) Fax: 650-3681101 jcai-at-nanostellar.com *********************************************************
From MicroscopyL-request-at-ns.microscopy.com Sun Dec 5 05:43:55 2004
I'm not sure this will work, but there is a new technology from Aetos called "CytoViva" which drops the limit of resolution of the light microscope from 250nm to below 120nm. It might give you just the edge you need.
Their website is www.CytoViva.com. Their tech apps person is Tom Hasling. I'm sure he'd be happy to run a sample for you. The website has all of their contact info.
Hope this was helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
At 07:43 AM 12/4/2004, somayyeh_kheiri-at-yahoo.com wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sun Dec 5 08:41:05 2004
Currently, I am organizing and chair a conference with title topic Vision'05 June 20-23, 2005 at LA, USA. The following is the detailed. Interested please kindly email you full paper to the following address:- ksism-at-mmu.edu.my or sksbg2003-at-yahoo.com
CALL FOR PAPERS
The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org
Title of Approved Session: Microscopy Imaging System
Chair of Session: Dr. Kok-Swee Sim Multimedia University, Melaka, Malaysia
Conference: The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org Monte Carlo Resort, Las Vegas, Nevada, USA June 20-23, 2005
Description: In this session, conference papers that submitted are related to Microscopy imaging system. The papers can be in the area of Material science microscopy and microanalysis, Medical applications of scanning microscopy, Microanalysis in Scanning Electron Microscope (SEM), Microscopy and microanalysis: theory, instrumentation and techniques, Monte Carlo modeling for microscopy and microanalysis, Multidimensional microscopy, Museum applications, Nanotechnology and nanofabrication, Scanning probe microscopies, Semiconductor devices, materials and process characterization, X-ray mapping in electron beam instruments.
Dear Colleagues: You are invited to submit a draft paper (see instructions below) and/or a proposal to organize a technical session/workshop. All accepted papers will be published in the conference proceedings.
The International Multiconference in Computer Science and Computer Engineering is a major annual research event. It assembles a spectrum of affiliated research conferences into a coordinated research meeting held in a common place at a common time. This model facilitates communication among researchers in different fields of computer science and computer engineering. The last Multiconference attracted over 1,650 computer science and engineering researchers from 78 countries. We expect to have over 2,000 attendees for this set of conference.
Please regard this announcement as General Guidelines. You are requested to send your submission to the chair whose address appears below.
Dr. Kok-Swee, Sim kssim-at-mmu.edu.my or sksbg2003-at-yahoo.com Tel: (+606) 252-3044 Fax: (+606) 231-6552 E-mail: sksbg2003-at-yahoo.com
SUBMISSION OF PAPERS:
Prospective authors are invited to submit three copies of their draft paper (about 5 pages - single space, font size of 10 to 12) to KS Sim by the due date (who may be forwarding the papers to respective conference chairs/committees). E-mail submissions in MS document or PDF formats are preferable (Fax submissions are also acceptable.)
The length of the Camera-Ready papers (if accepted) will be limited to 7 (IEEE style) pages. Papers must not have been previously published or currently submitted for publication elsewhere. The first page of the draft paper should include: title of the paper, name, affiliation, postal address, E-mail address, telephone number, & Fax number for each author. The first page should also include the name of the author who will be presenting the paper (if accepted) & a maximum of 5 keywords.
IMPORTANT DATES:
Feb. 16, 2005: Submission of papers (about 5 pages) March 21, 2005: Notification of acceptance April 20, 2005: Camera-Ready papers & Prereg. due June 20-23, 2005: The 2005 World Congress in Applied Computing (WCAC'05) - 14 Joint Conferences.
Thanks Ks
From MicroscopyL-request-at-ns.microscopy.com Sun Dec 5 14:26:13 2004
Can anyone give me the current contact information for whoever is likely to have replacement parts for a 20year old LKB knifemaker? I need to replace the scoring wheel in ours.
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 Mon Dec 6 07:31:04 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rvf263-at-chartermi.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, December 5, 2004 at 21:42:43 ---------------------------------------------------------------------------
Email: rvf263-at-chartermi.net Name: Robert Farris
Organization: Monroe County Community College
Education: Undergraduate College
Location: Newport, Michigan - United States
Question: I'm taking Biology in college now, will be taking Anatomy & Physiology this winter, and Microbiology soon. Can you recommend a decent microscope for me to purchase for use at home? Thank you very much Robert
We are looking to purchase a new high-vacuum carbon evaporation unit for our lab. I have received information on the Cressington 208 and 308 models, the Denton Bench Top Turbo IV, and Emitech.
Does anyone have any experience with these units? Can anyone comment on their reliability and performance? Anyone have any other recommendations for coaters?
The coater will be used for SEM/EDS analysis of samples with topography. We mainly work on inorganic materials. It may also, in the future, be used for TEM work.
Thank you in advance for your help. Amy Bern
^v^ Amy M. Bern Chemist EPA, NEIC, Bldg. 25 P.O. Box 25227 Denver, CO 80225 Phone: 303-462-9128
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 6 14:00:46 2004
Debby, At Ted Pella Inc, we offer an overhaul service on LKB knifemakers for $500 plus shipping. This includes a new scoring wheel and other consumable parts. Check it out at: http://www.tedpella.com/glass_html/knifemkr.htm We do not offer the parts separately.
Regards, Mark Armogida VP of Engineering and Production
Phone: 530.243.2200 X212 Fax: 530.243.3761
Ted Pella, Inc. www.tedpella.com
-----Original Message----- } From: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Sunday, December 05, 2004 12:54 PM To: message to: MSA list
I am looking for glue for embedding samples (materials and minerals). Please advise the glue that is suitable for materials. EpoFix is out of stock at Electron Microscopy Sciences, so I am looking for a similar one from the other companies. I am embedding samples for ultramicrotomy.
Thank you, Hiromi Konishi, Ph.D. Indiana University
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 6 23:33:02 2004
} We recondition LKB Knife Makers, including scoring wheel etc. for $250.00 plus shipping. Turn-around 1 week. We also recondition Sorval MT-1, MT-2 and 2B Ultra Microtomes and sell belt kits/ motor mounts for MT-2/2B. Regards Markus F. Meyenhofer Microscopy Labs Box 338 61 West Street Red Bank, NJ 07701 732 747 6228 fax 732 758 9142 } } } ---------------------------------------------------------- } -------------------- The Microscopy ListServer -- Sponsor: } The Microscopy Society of America To
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 7 07:56:11 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stacey.Andringa-at-uc.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, December 6, 2004 at 13:32:34 ---------------------------------------------------------------------------
To be held in conjunction with the March 24,2005 meeting of the Midwest Microscopy and Microanalysis Society
Affiliate of the Microscopy Society of America and the Microbeam Analysis Society of America
!!PLEASE NOTE EXTENDED DEADLINE FOR RECEIPT OF ABSTRACTS!!
ABSTRACT DEADLINE: Abstracts must be received in electronic format by 5PM, WEDNESDAY, DECEMBER 22, 2004.
The first quarter meeting of the Midwest Microscopy and Microanalysis Society will be held on Thursday, March 24, 2005, in the Pancoe Life Sciences Building, Northwestern University, Evanston, Illinois. A student poster competition open to undergraduate and graduate students will be held in conjunction with the meeting. Posters should illustrate utilization of microscopy for either biological or materials science study. Prizes will be awarded as follows:
$300 for 1st place $200 for 2nd place $100 for 3rd place
Abstracts must be received in electronic format by 5PM on Wednesday, December 22, 2004. To be eligible for a prize, you must be first author on the poster, and you must be present at the meeting. You are encouraged to submit your entry as early as possible, as space may be limited. Further details and an explanation of judging criteria will be provided upon acceptance of your submission.
Please send the following information to the email address below, and attach your abstract as a Word document:
Name Affiliation Department Phone number Email address
Send to:
Elaine Schumacher MMMS Materials Science Director McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 Tel: 630-887-7100 Fax: 630-887-7417 Email: eschumacher-at-mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 7 13:47:17 2004
ElectroscanE3 for sale as a standing unit or we will part it out. This ESEM is a working unit though the image quality is poor right now either due to the scintillator or the column is dirty. Hit stage, tensile stage, and peltier stage; LaB6 ready.
No offers for the standing unit under $10K, if you need parts we will discuss the price on an as need basis. Water chiller not included.
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 7 21:41:38 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rebecca.burgin-at-onsemi.com) from http://www.msa.microscopy.org/MicroscopyListserver/MLFormMail.html on Tuesday, December 7, 2004 at 17:26:50 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fchu-at-mrl.ubc.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 7, 2004 at 16:25:24 ---------------------------------------------------------------------------
Email: fchu-at-mrl.ubc.ca Name: Fanny Chu
Organization: iCAPTURE Lab, University of British Columbia
Question: Somebody in our lab has had sections that will get diffused and dispersed within two seconds staying in the boat. Ironically, all the Epon ingredients were brand new. Any suggestions what's wrong or can be done about it?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (drbenjy-at-optonline.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, December 7, 2004 at 22:36:24 ---------------------------------------------------------------------------
Email: drbenjy-at-optonline.net Name: Ben Glassman, MD
Organization: Physician in private practice
Education: Graduate College
Location: Mamaroneck, New York, USA
Question: I have a number of mounted blood smears of unusual anemias. The mountant is yellowing and would like to know if it is possible to salvage these slides. Also ,is it possible to restain them
Fanny We've all been hit with this phenomenon at some time ...It sounds like incomplete infiltration and/or polymerization to me. Even with brand new components, if they are not mixed adequately the resin won't harden correctly. Also, if the dehydration was not complete (your absolute ethanol or acetone was not actually absolute) the resin will not be able to fully infiltrate the tissue and you will see the effect you've described. I'm afraid there is no salvaging of those blocks (at least none that I've found). Next time, make sure that you use a fresh bottle of 100% dehydrating agent, and also take the time to thoroughly mix the resin components. It may even help to stir them up, let them sit for 15-30 minutes and stir them again. We used to do that with Spurr's resin all the time. 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 Wed Dec 8 07:42:30 2004
} Question: I have a number of mounted blood smears of unusual } anemias. The mountant is yellowing and would like to know if it is } possible to salvage these slides. Also ,is it possible to restain } them } } --------------------------------------------------------------------------- the histologist who shares my lab space routinely rescues slides from the medical students' slide collection. She soaks them in a coplin jar of xylene overnight, or longer, until the old cover glass falls off. She then restains them if needed and mounts a new cover glass with Permount. She usually handles paraffin sections and they come through this procedure very nicely. I'm not sure if blood smears would restain well. I would try it with you least valuable slide first as a test specimen.
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 Wed Dec 8 08:39:05 2004
Any time your section diffuse during microtomy, it is usually caused by incomplete infiltration of the tissue. This will occur even if your epon ingredients are new. You must draw out your infilitration steps, ie: from 2 x 5 min. propylene oxide, 3:1 P.O.: epon for 2 hrs then 1:1 P.O.: Epon 2 hrs or overnight, 1:2 P.O.: Epon 2 hrs, then 3 fresh Epon changes. Vacuum infiltration also helps, and Spurrs is a better infiltrating resin. Good Luck.
Michael Delannoy Assist. Director Johns Hopkins School of Medicine Microscope Facility
----- Original Message ----- } From: fchu-at-mrl.ubc.ca (by way of MicroscopyListserver)
Hello All, I am posting this ad for Prof. Yang. Please look at the detail below.
The Department of Materials Science and Engineering at the University of Pittsburgh seeks a post-doctoral research associate for in situ transmission electron microscopy studies of surface oxidation reactions of metallic alloys. The successful applicant will utilize primarily a unique in situ ultra-high vacuum transmission electron microscope that resides within the Materials Research Laboratory at the University of Illinois at Urbana-Champaign. The appointment is initially for one year with possible extension, starting immediately.
Surface oxidation processes play critical roles in environmental stability, corrosion, electrochemistry, catalysis, gate oxides, fuel reactions, as well as nanostructures and thin films formed by surface reactions. Furthermore, as engineered materials approach the nanometer scale, environmental stability at this scale is critical to the device performance. The objective of this research program is fundamental understanding of nano-oxidation reactions via a coordinated experimental effort, with in situ UHV-TEM (University of Pittsburgh) and synchrotron X-ray diffraction (Argonne National Laboratory), and theoretical and simulations effort (University of Florida). The experimentalists and theorists will interact closely.
A Ph.D. in Materials Science and Engineering, Physics, or related field is necessary. Required experience includes transmission electron microscopy. Experience with in situ, advanced electron microscopy methods and/or other characterization methods, such as AFM, and thin film deposition is advantageous, but not required.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang
848 Benedum Hall
Materials Science & Engineering Dept.
University of Pittsburgh
Pittsburgh, PA 15261
jyang-at-engr.pitt.edu
(412) 624-8613
Long Li _________________________________________________________ Materials Science and Engineering Department University of Pittsburgh 3700 O'Hara St., 848 Benedum Hall Pittsburgh, PA 15261
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gautam-at-jncasr.ac.in) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, December 8, 2004 at 10:01:24 ---------------------------------------------------------------------------
Email: gautam-at-jncasr.ac.in Name: Gautam
Organization: JNCASR
Education: Graduate College
Location: Bangalore, Karnataka, India
Question: Hi. Is it possible that we see a doubling of the lattice spacing in a high-resolution electron microscopy TEM image. If yes, what are the reasons and could you suggest some references?
This depends on the numerical aperture/resolving power of the objective lens being measured. Ten microns is almost certainly too big though. To generate a point-spread function dataset you will need to use beads that are well below the resolution limit of the lens in question. Molecular probes sells such beads under the brand name "Tetraspeck" (140nm). Polysciences sells very small fluorescent beads as well (down to 50nm). Quantum dots (~10nm) are another alternative, however they are somewhat expensive. -Karl
Peter Lundh wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 9 10:26:49 2004
Does anyone has the experience on working edge detection imaging technique on SEM images ? Just curious also, what is the implication of applying edge detection technique such as canny, sobel and so on onto SEM images.
If anyone has the idea, please kindly share with me.
Many thanks Kok Swee
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 9 10:28:05 2004
When I prepare my Epon resin I stir two components together for 5-10 min, add the third, stir for another 5-10 min before adding the accelerator and stir for 30 min. I get consistent result this way. If you put all four together and stir, you may get various results.
Ann Fook Yang
-----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: Wednesday, December 08, 2004 9:05 AM To: by way of MicroscopyListserver; microscopy-at-microscopy.com
Fanny We've all been hit with this phenomenon at some time ...It sounds like incomplete infiltration and/or polymerization to me. Even with brand new components, if they are not mixed adequately the resin won't harden correctly. Also, if the dehydration was not complete (your absolute ethanol or acetone was not actually absolute) the resin will not be able to fully infiltrate the tissue and you will see the effect you've described. I'm afraid there is no salvaging of those blocks (at least none that I've found). Next time, make sure that you use a fresh bottle of 100% dehydrating agent, and also take the time to thoroughly mix the resin components. It may even help to stir them up, let them sit for 15-30 minutes and stir them again. We used to do that with Spurr's resin all the time. 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 Dec 10 07:22:59 2004
I am currently organizing and chair an session for USA conference Vision'05. Please kindly support by sending me conference paper.
Many thanks The following is the information:-
CALL FOR PAPERS
The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org
Title of Approved Session: Microscopy Imaging System
Chair of Session: Dr. Kok-Swee Sim Multimedia University, Melaka, Malaysia
Conference: The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org Monte Carlo Resort, Las Vegas, Nevada, USA June 20-23, 2005
Description: In this session, conference papers that submitted are related to Microscopy imaging system. The papers can be in the area of Material science microscopy and microanalysis, Medical applications of scanning microscopy, Microanalysis in Scanning Electron Microscope (SEM), Microscopy and microanalysis: theory, instrumentation and techniques, Monte Carlo modeling for microscopy and microanalysis, Multidimensional microscopy, Museum applications, Nanotechnology and nanofabrication, Scanning probe microscopies, Semiconductor devices, materials and process characterization, X-ray mapping in electron beam instruments.
Dear Colleagues: You are invited to submit a draft paper (see instructions below) and/or a proposal to organize a technical session/workshop. All accepted papers will be published in the conference proceedings.
The International Multiconference in Computer Science and Computer Engineering is a major annual research event. It assembles a spectrum of affiliated research conferences into a coordinated research meeting held in a common place at a common time. This model facilitates communication among researchers in different fields of computer science and computer engineering. The last Multiconference attracted over 1,650 computer science and engineering researchers from 78 countries. We expect to have over 2,000 attendees for this set of conference.
Please regard this announcement as General Guidelines. You are requested to send your submission to the chair whose address appears below.
Dr. Kok-Swee, Sim kssim-at-mmu.edu.my or sksbg2003-at-yahoo.com Tel: (+606) 252-3044 Fax: (+606) 231-6552 E-mail: sksbg2003-at-yahoo.com
SUBMISSION OF PAPERS:
Prospective authors are invited to submit three copies of their draft paper (about 5 pages - single space, font size of 10 to 12) to KS Sim by the due date (who may be forwarding the papers to respective conference chairs/committees). E-mail submissions in MS document or PDF formats are preferable (Fax submissions are also acceptable.)
The length of the Camera-Ready papers (if accepted) will be limited to 7 (IEEE style) pages. Papers must not have been previously published or currently submitted for publication elsewhere. The first page of the draft paper should include: title of the paper, name, affiliation, postal address, E-mail address, telephone number, & Fax number for each author. The first page should also include the name of the author who will be presenting the paper (if accepted) & a maximum of 5 keywords.
IMPORTANT DATES:
Feb. 16, 2004: Draft papers (about 5 pages) due March 22, 2004: Notification of acceptance April 21, 2004: Camera-Ready papers & Prereg. due June 21-24, 2004: 2004 Int'l Multiconference in CS & CE
----- Original Message ----- } From: "by way of MicroscopyListserver" {jean-paul.bailon-at-polymtl.ca} To: {microscopy-at-microscopy.com} Sent: Tuesday, November 30, 2004 7:13 AM
Dear All,
I am currently organizing and chair an session for USA conference Vision'05. Please kindly support by sending me conference paper.
Many thanks The following is the information:-
CALL FOR PAPERS
The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org
Title of Approved Session: Microscopy Imaging System
Chair of Session: Dr. Kok-Swee Sim Multimedia University, Melaka, Malaysia
Conference: The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org Monte Carlo Resort, Las Vegas, Nevada, USA June 20-23, 2005
Description: In this session, conference papers that submitted are related to Microscopy imaging system. The papers can be in the area of Material science microscopy and microanalysis, Medical applications of scanning microscopy, Microanalysis in Scanning Electron Microscope (SEM), Microscopy and microanalysis: theory, instrumentation and techniques, Monte Carlo modeling for microscopy and microanalysis, Multidimensional microscopy, Museum applications, Nanotechnology and nanofabrication, Scanning probe microscopies, Semiconductor devices, materials and process characterization, X-ray mapping in electron beam instruments.
The International Multiconference in Computer Science and Computer Engineering is a major annual research event. It assembles a spectrum of affiliated research conferences into a coordinated research meeting held in a common place at a common time. This model facilitates communication among researchers in different fields of computer science and computer engineering. The last Multiconference attracted over 1,650 computer science and engineering researchers from 78 countries. We expect to have over 2,000 attendees for this set of conference.
Please regard this announcement as General Guidelines. You are requested to send your submission to the chair whose address appears below.
Dr. Kok-Swee, Sim kssim-at-mmu.edu.my or sksbg2003-at-yahoo.com Tel: (+606) 252-3044 Fax: (+606) 231-6552 E-mail: sksbg2003-at-yahoo.com
SUBMISSION OF PAPERS: Prospective authors are invited to submit three copies of their draft paper (about 5 pages - single space, font size of 10 to 12) to KS Sim by the due date (who may be forwarding the papers to respective conference chairs/committees). E-mail submissions in MS document or PDF formats are preferable (Fax submissions are also acceptable.)
The length of the Camera-Ready papers (if accepted) will be limited to 7 (IEEE style) pages. Papers must not have been previously published or currently submitted for publication elsewhere. The first page of the draft paper should include: title of the paper, name, affiliation, postal address, E-mail address, telephone number, & Fax number for each author. The first page should also include the name of the author who will be presenting the paper (if accepted) & a maximum of 5 keywords.
Feb. 16, 2005: Submission of papers (about 5 pages) March 21, 2005: Notification of acceptance April 20, 2005: Camera-Ready papers & Prereg. due June 20-23, 2005: The 2005 World Congress in Applied Computing (WCAC'05) - 14 Joint Conferences.
----- Original Message ----- } From: "by way of MicroscopyListserver" {jean-paul.bailon-at-polymtl.ca} To: {microscopy-at-microscopy.com} Sent: Tuesday, November 30, 2004 7:13 AM
Listers,
The Alcatel MDP-5010 molecular drag pump that backs the turbo pump on our S-900 has died, and the 5010 can no longer be rebuilt. Alcatel has quit making or providing parts. So, I'm wondering if anyone has an old Alcatel MDP-5010 lurking around in working condition (preferably rebuilt, controller not needed), or has any extra 7+ cfm (170+ L/min) oil-free scroll pumps, like the Edwards XDS-10. Which latter are rarer than hens' teeth, it seems. Thanks for any help.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 11 07:48:44 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Springpaard-at-yahoo.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, December 9, 2004 at 15:34:20 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (shuckaby-at-reddyus.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, December 9, 2004 at 14:40:56 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] Use of Alcian Blue for staining of proteoglycans
Question: I hope some one can help me out here. I am trying to use Alcian Blue staing to quantitate matrix proteoglycans. Currently the system i have worked out is to grow tissue culture cells of interest on glass cover glass, fix the cells to the cover glass in 4%formalin/PBS for 20 minutes washed in ddH2O and then stain with 1% Alcian blue(w/v) solution prepared in 3%acetic acid (in water). I allow the staining to progress for 2 hours at room temperature then remove the staining solution, wash the coverglass with the fixed and stained cells and mount onto the slides. I am using a Nikon light microscope and 100x oil imersion lense to see the actual extracellular matrix of the cells. The problem that I am having is that the staining is not very even and I have what look to be large crystals of the blue that are not washed off. I am looking to reduce background due to these large crystals so I can just see nice green-blue matrix. I have tried filtering the dye solution with minimal success. If you have any tips for staining proteoglycans in tissue culutre with Alcian blue please let me know. THANKS!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kssim-at-mmu.edu.my) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, December 9, 2004 at 10:26:21 ---------------------------------------------------------------------------
Does anyone has the experience on working edge detection imaging technique on SEM images ? Just curious also, what is the implication of applying edge detection technique such as canny, sobel and so on onto SEM images.
If anyone has the idea, please kindly share with me.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stacey.Andringa-at-uc.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, December 10, 2004 at 10:04:40 ---------------------------------------------------------------------------
Question: I have a co-worker looking for a used microtome part. It is a standard fixed jaw specimen clamp for an RMC MT-980 (now designated MR3)rotary microtome. I have done a google search with little success. If anyone knows a good source for used parts, please let me know. Thanks.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (qizhang-at-physics.unc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, December 10, 2004 at 16:10:29 ---------------------------------------------------------------------------
Our JEM-2010F microscope has beam oscillating with 2-3 Hz frequency now. I couldnít say when it started. But we didnít have this problem before. Then we took out the gun chamber several times to open the column for mechanical alignment and the polepiece replacement. Now, the voltage stability and emission stability are OK. Does anyone have had this problem or have any comment and suggestion on troubleshooting? Any comment and suggestion will be highly appreciated! Thank you very much!
Regards, ************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 (Office) 919-843-0974 (EM Lab) Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (balbrecht-at-nwicc.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, December 10, 2004 at 12:46:18 ---------------------------------------------------------------------------
Email: balbrecht-at-nwicc.edu Name: Brian Albrecht
Organization: Northwest Iowa Community College
Education: Undergraduate College
Location: Sheldon, Iowa, United States
Question: I'm a science professor here at Northwest Iowa Community College. Our welding instructor, through the Department of Energy, obtained a Leitz Metallurgy microscope. No instructions came with the microscope. We are looking for a way of obtaining an instruction manual. The microscope does not have standard adapters for plugging into the wall. The adapter ends consist of a cylinder which house I believe 5 flat prongs probably no longer than a half an inch and no wider than probably a quater of an inch. The only number that was visible on the scope was 741390. Any assistance would be greatly appreciated. Thank you.
We would like to service and test an old Omar cpd after a couple of years in a box. Is there a company or facility repairing/servicing/testing such a unit?
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 08:16:32 2004
It seems that the $10k asking price for the Electroscan was too rich for the list. We are willing to take $5K from anyone willing to remove it and enjoy the wonders of LVSEM! Don't forget about the HT stage, tensile stage and peltier stage; you can do almost anything with this tool. And if you just need parts for your ESEM $5K is really fair. Scrap metal prices are really high now too, we just may take that route if someone doesn't make an offer, so act now! What about that hard to buy for Uncle this holiday season? Wouldn't he be surprised when you drop this off on his lawn? The old guy needed a hobby anyway. The possibilities are endless, act now or off to Finger Lakes Scrap with it!
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 09:04:09 2004
Greetings, Total internal reflectance microscopy (TIRF) provides resolution on the order of tens of nanometers in the axial dimension. This is because of a a phenomenon which occurs when the rays of excitation illumination are glanced off of a reflective interface (an interface between differing refractive indices) at an angle greater than the second refractive index will accomodate (in other words, a much higher numerical aperture than is possible to attain in the second refractive index). When light is reflected off of an interface in this manner, a very small portion of the media past the second refractive interface is illuminated by evanescent waves from the illumination source. The intensity of evanescent waves decays very rapidly so illumination is only intense enough to excite fluorochromes to a depth of a few tens of nanometers. This provides a means of determining that bright spots are localized to very near the interface with a resolution which surpasses that of conventional diffraction limited illumination schemes. An example of this would be viewing the distribution of labeled proteins in the cell membrane of cultured cells adhering to the coverslip. To answer your question more directly, the big advantage of TIRF is very high depth resolution at the coverslip/sample media interface. That is the single big advantage. This means a very small sample volume can be monitored and investigators can monitor molecules moving into and out of this small volume. From these observations, calculations which provide insight into molecular concentrations, whether or not the movement of molecules appears to be random, correlation of spatial relationships between (and colocalization among) different types of molecules and other parameters can be conducted. Cheers, Karl
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 09:10:17 2004
The Laboratory of Developmental Genetics at The Rockefeller University seeks a full-time electron microscopist to assist in studies of nervous system development and cell death in the nematode C. elegans.
Work will involve preparing specimens for TEM, serial sectioning, and 3-D image reconstructions. The successful candidate will become an integral part of ongoing scientific efforts in the lab, and could also pursue independent research. Previous experience with TEM is required, and experience with serial sectioning experience a plus. Applicants must hold a minimum of a Bachelor's degree.
We offer a competitive salary, comprehensive benefits, a gracious working environment, and tuition reimbursement. For immediate consideration, send resume/C.V. to: Ms. Kara R. Marshak, Senior Employment Specialist, The Rockefeller University via fax (212)-327- 7079 or email marshak-at-rockefeller.edu. For more information about ongoing research in this laboratory,please visit our website located at http://www.rockefeller.edu/labheads/shaham/shaham-lab.php.
An Affirmative Action/Equal Opportunity Employer. The Rockefeller University appreciates all responses and will contact candidates selected for further consideration.
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 10:54:29 2004
I never thought it would happen here but the prospect of inter -departmental cross charging is being now being discussed. Does anyone have recent experience of constructing a price list for; Processing samples? for immunolabelling ? etc etc. Where is the best place to start? I am not sure if we will have to charge to cover consumables or entire costs.
Ken Blight Senior Scientific Officer Cancer Research UK
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 12:17:24 2004
Greetings, I'm looking for an incident fluorescence illumination system to retrofit an old Olympus BH-2 stand. Any input as to where I might find such parts in good conditions would be appreciated. Thanks. Regards, Karl
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 15:25:50 2004
I had to do this from scratch when I started here 5 years ago. It can be quite simple. First, estimate hourly fee for technical help by dividing average salary of technician plus benefits by a reasonable estimate of billable hours that a technician can provide. Don't be over-optimistic - only a few hours a day can actually be billed for. A lot of duties that cannot be accounted for (such as administrative work, ordering, cleaning the lab up, teaching, answering information requests, lab meetings, etc...). We came up with a number of $40 an hour, but this is in fact too low and we will review it upwards soon (should be at least $50). Then estimate average amount of time required for the tasks you want to start billing (for example: cryo- sectioning / 1hour per sample - Immuno-labeling / 2 hours per run, etc), add some cost for consumables (liquid nitrogen, coated grids, antibodies, ...) and you can estimate a reasonable charge.
For sectioning, you also need to take into consideration the cost for diamond knives. Depending on the experience of you staff (and the nature of the samples you will be cutting), estimate how many samples in average you will be sectioning with one diamond knife before it needs resharpening. People have different opinions about when a knife needs resharpening, but in my view if you are going to sell your sections at a hefty price, they have to be perfect (NO knife marks!). We estimate a cost of about $5 per sample just for the knife when we do the cutting, and charge users $20 per hour for knife when they cut themselves (sometimes literally!!). See it as a form of insurance.
For instrument rental, we calculate a hourly rate by dividing service contracts plus technician time required for routine maintenance, by the total number of hours the instrument will be used in the year (by our staff or outside users).
Some of our current charges: Technician time: $41.50/hour EM time (unassisted): $41.50/hour EM time (assisted): $83/hour Microtome rental: $15.50/hour Cryo-microtome rental: $26/hour Embedding: $36.50/sample Sectioning: $52/sample Cryo-sectioning: $93.50/sample Single immuno-labeling: $125/run Double immuno-labeling: $230/run Prints: $6.50 EM negative: $1.60
We also have a "comprehensive" charge for regular or immuno-EM, that comprises every step from fixation to the production of publication-quality pictures. The cost is $155 for regular EM, and $310 for immuno-EM (per sample).
Hope this helps.
Marc
On Monday, December 13, 2004, at 12:24 PM, Ken Blight 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 never thought it would happen here but the prospect of inter } -departmental cross charging is being now being discussed. Does anyone } have recent experience of constructing a price list for; Processing } samples? for immunolabelling ? etc etc. Where is the best place to } start? I am not sure if we will have to charge to cover consumables or } entire costs. } } Ken Blight } Senior Scientific Officer } Cancer Research UK } }
-- 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 Mon Dec 13 20:30:41 2004
I'm trying to write a batch converter that will convert a Veeco format image to something more universal, namely TIFF in ImageJ. Does anyone know the compression used or has anything been done like this before?
Thanks Thomas Sadowski Southern Connecticut State University
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 06:42:06 2004
I would like to know how I can find abstract of Meeting of Microscopy & Microanalysis. I use Web of Science. However, I cannot find my abstracts. It seems that Web of Science do not have information on Microscopy & Microanalysis meeting. What database has the information? Please advise.
Thank you, Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 08:30:45 2004
This subject has been discussed a few times here. When I saw the text below in my latest NASA Tech Briefs INSIDER, I thought it may be of interest to at least one of the listers, so copied it and am sending it along. The copyright and subscription info is below, also. Enjoy...
DATA STORAGE Will records stored on CD or DVD still be retrievable 10 or 20 years from now? Researchers at the National Institute of Standards and Technology (NIST) tested how well recordable optical disks made with different manufacturing processes held up to high temperatures, humidity, and light levels. Results showed that some disks perform better than others and that excessive exposure to any of these conditions can accelerate deterioration.
To address how to identify those high-quality media for archival applications, NIST, along with the DVD Association (DVDA) and several government agencies, has formed the Government Information Preservation Working Group.
Working with the optical disk industry, the group is setting requirements for archival quality CD and DVD recordable media, and specifying the minimum number of years that they must last to meet the requirements. NIST is also developing a test that manufacturers can use to determine if CDs and DVDs meet the criteria.
Read the complete story at: http://link.abpi.net/l.php?20041213A6
Please let your colleagues know they too can receive the INSIDER free of charge simply by sending an e-mail message to the address Listserv-at-listserv.abpi.net with the text SUBSCRIBE Insider Firstname Lastname as the only text on the first line of the message body.
For information on how your company can sponsor future editions of the INSIDER, e-mail joe-at-abpi.net .
Copyright (c) 2004 Associated Business Publications Intl.
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 12:17:43 2004
} This subject has been discussed a few times here. ...
It's still an open subject ...
} Read the complete story at: http://link.abpi.net/l.php?20041213A6
There is a PDF available if you follow the hyperlinks at the above URL ... "Stability Comparison of Recordable Optical Discs - A Study of Error Rates in Harsh Conditions". However, the disappointing point made in the summary is:
"It is demonstrated here that CD-R and DVD-R media can be very stable (sample S4 for CD-R and sample D2 for DVD-R). Results suggest that these media types will ensure data is available for several tens of years and therefore may be suitable for archival uses. Unfortunately, it is very difficult for customers to identify these more stable media."
Recognizing the better media sometimes requires software to know where the media was manufactured. Another good source for manufacturer info can be explored here: http://www.digitalfaq.com/media/dvdmedia.htm
I have a Leitz Ergolux microscope with a broken focus mechanism. It's at a repair facility in Texas. The repair company says that Leica's parts website reports that they no longer stock the two gears need for the repair of my scope since the scope is at least 15 years old.
So I'm exploring two routes:
1. Find a used Ergolux body and retrieve the parts I need or swap my objectives, head, stage, illuminator, etc. to the new body and use it instead.
2. Either buy of-the-shelf gears from a gear parts company or have custom gears machined to repair the Ergolux. The second part of this proposition is the most expensive as it's custom, only yields the two gears I need while a replacement body might hold other parts for future repairs. I've talked to a gear manufacturing company in town (Phoenix) and they're saying to reverse engineer the gears I need from scratch might be in the $500 range, which is more than I want to pay for two gears less than 1" (25mm) in size.
Do any of you have a used Ergolux body that's collecting dust that you'd sell for a reasonable price?
Do any of you know if any other Leitz scopes have a matching focus mechanism where the gears can be interchanged with an Ergolux?
Or do you know of a gear parts company you can recommend that might stock what I need?
Thanks for your help.
Doug Baldwin
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 15:19:18 2004
We are in urgent need of two boards for our Hitachi H7000 Transmission Electron Microscope. Both boards are lens board. One board is called lens STB, part no. 747-0708 and the other one is DC power supply PC3 (don't know the part number).
If someone has the boards and is willing to sell them to us please contact me off line as soon as possible. The scope has been down for about a month now.
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://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 16:49:28 2004
I know finding funding for new instruments is very competative, so if you don't want to give away any secrets, I understand.
But, I am stuck. Folks are coming into my little multiuser lab and asking how we can get some new equipment. None of them individually present the critical mass needed to justify a microscope, but all together maybe they have a point.
I have looked around and know about the NSF MRI program, but we missed the deadline for this year and our campus may have different priorities for the limited number of submissions they can do.
I know about the various NSF equipment programs, has anyone tried to do this using PI's from multiple disciplines? I have users from Biology, Earth Science, Chemistry, maybe some others, none which could do it on their own, but together, maybe.
How about any other leads? As I said, don't give away the farm, but maybe you could clue me in a little.
Thanks
Jon
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 21:33:49 2004
We have a researcher who would like to do some transmission electron microscopy on synovial fluid from the knee. He is looking for debris in the fluid after a particular surgical procedure. this debris will not necessarily be cellullar, it may be from the superficial cartilage layer.
We thought of either spinning the fluid down and processing the pellet that may result, or perhaps smearing the fluid onto a coated slide, fixing it and then doing the pop off technique.
Either way I imagine the amount of organic matter we are going to get will be pretty small.
Has anybody out there got any suggestions or experience with the transmission electron microscopy of 'clear' body fluids, ie synovial fluid. I imagine cerebral spinal fluid would be pretty similar in organic matter content.
I am not sure if we are going to run into problems with high protein content in these fluids, perhaps masking the debris this researcher is actually looking for, any thoughts?
Many thanks
Allan
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 06:08:15 2004
A SEM we are purchasing has an application that does not require optimum gun brightness, and for the sake of filament longevity (months of 24-7) runs the filament heat at less than saturation and on top of the 1st peak.
I have always tought that the phenomenon of the 1st peak must be a result of the Wehnelt's geometry as a function of heating and the zero-to-positive keV potential moving towards the filament tip ... that is, a phenomenon of note but not much more of a concern. Now, I find myself wanting of a better understanding for the sake of beam current stability ... i.e., operating on the first peak does not seem to be a very stable parameter for stable emission.
The SEM itself is a FEI Quanta, which apparently has a method of regulating the emission current. This must also play a role, and I haven't yet found a explanation of how this works.
Any discussion towards enlightenment much appreciated.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings
I know finding funding for new instruments is very competative, so if you don't want to give away any secrets, I understand.
But, I am stuck. Folks are coming into my little multiuser lab and asking how we can get some new equipment. None of them individually present the critical mass needed to justify a microscope, but all together maybe they have a point.
I have looked around and know about the NSF MRI program, but we missed the deadline for this year and our campus may have different priorities for the limited number of submissions they can do.
I know about the various NSF equipment programs, has anyone tried to do this using PI's from multiple disciplines? I have users from Biology, Earth Science, Chemistry, maybe some others, none which could do it on their own, but together, maybe.
How about any other leads? As I said, don't give away the farm, but maybe you could clue me in a little.
Thanks
Jon
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 07:55:43 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Ntores-at-btconnect.com) from on Wednesday, December 15, 2004 at 05:29:58 ---------------------------------------------------------------------------
Email: Ntores-at-btconnect.com Name: Nicholas Tores
Organization: IPS
Title-Subject: [Microscopy] [Filtered] Fofler heating bench ( Heizbank German )
Question: We are looking to purchase a Kofler Hotbench for thermal treatment of tiny specimens. Can you help ?
I received some nematodes for SEM. They are preserved in 96% ethanol. I have no experience with this sort of organisms. Can anyone please tell me how to proceed?
Jon, The Core Facility Management at M&M 2004 dealt with this topic. Representatives from NSF and NIH spoke on funding opportunities for major equipment acquisition. It turns out that there are good opportunities for both PhD granting and non-PhD granting institutions.
The session was taped and transcribed and the first segment (NIH opportunities) will be published in Microscopy Today in the January issue. Other segments will appear over the next few issues. Included will be Questions & Answers that give a lot of suggestions to help make your grant more competitive.
The NIH NCRR Major Equipment grant deadline is mid-March so you will see that article in plenty of time. NSF deadlines for the MUE program (biology) and the IMR (Instrumentation for materials Research) are in early October.
One thing that was strongly encouraged was to call the NSF program director where you think you would best fit and talk to that individual. They can guide you as to what division would be most appropriate for your proposal and assist in answering many other questions regarding writing a successful proposal.
One new development that you may not be aware of is that NSF is dropping the required 30% match for major equipment proposals. They are also increasing the maximum for the MUE and IMR programs from $400,000 to $500,000. This should help a lot when you try to round up funds for the more expensive instruments.
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 12/14/04 6:17 PM, "Jon Krupp" {jmkrupp-at-cats.ucsc.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 } ------------------------------------------------------------------------------} - } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------} - } } Greetings } } I know finding funding for new instruments is very competative, so if you } don't want to give away any secrets, I understand. } } But, I am stuck. Folks are coming into my little multiuser lab and asking } how we can get some new equipment. None of them individually present the } critical mass needed to justify a microscope, but all together maybe they } have a point. } } I have looked around and know about the NSF MRI program, but we missed the } deadline for this year and our campus may have different priorities for the } limited number of submissions they can do. } } I know about the various NSF equipment programs, has anyone tried to do } this using PI's from multiple disciplines? I have users from Biology, Earth } Science, Chemistry, maybe some others, none which could do it on their own, } but together, maybe. } } How about any other leads? As I said, don't give away the farm, but maybe } you could clue me in a little. } } Thanks } } Jon } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 09:32:23 2004
Bonjour everyone! I had a question from a prof I couldn't answer and was wondering if you could help. He has a couple of diamond knives he wants sharpened (Diatome). He asked me if he could get the work done in Canada. I have always sent my knives back to the manufacture. Has anyone out there ever gotten a knife sharpened in Canada and if yes, let me know about it....You could also contact me off line if you wish. Merci..
André Dufresne University of Manitoba Botany Dept. Dufresne-at-ms.umanitoba.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 09:54:59 2004
Another option would be to use a cytospin to concentrate the material on the slide; then invert a beem capsule over it and process for TEM.
Peggy Sherwood Lab Associate, Photopathology Wellman Center for 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: Allan Mitchell [mailto:allan.mitchell-at-stonebow.otago.ac.nz] Sent: Tuesday, December 14, 2004 11:04 PM To: microscopy-at-msa.microscopy.com
Kia Ora
We have a researcher who would like to do some transmission electron microscopy on synovial fluid from the knee. He is looking for debris in the fluid after a particular surgical procedure. this debris will not necessarily be cellullar, it may be from the superficial cartilage layer.
We thought of either spinning the fluid down and processing the pellet that may result, or perhaps smearing the fluid onto a coated slide, fixing it and then doing the pop off technique.
Either way I imagine the amount of organic matter we are going to get will be pretty small.
Has anybody out there got any suggestions or experience with the transmission electron microscopy of 'clear' body fluids, ie synovial fluid. I imagine cerebral spinal fluid would be pretty similar in organic matter content.
I am not sure if we are going to run into problems with high protein content in these fluids, perhaps masking the debris this researcher is actually looking for, any thoughts?
Many thanks
Allan
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 10:32:43 2004
If you go to our website at www.wormatlas.org you will see a listing for "Anatomical Methods" for the study of the nematode. In this listing, there is a detailed protocol for doing SEM on the nematode, or even on dissected organs from the nematode.
Other portions of our website will help to inform you about the various tissues that you might encounter, but you may especially want to read the "Cuticle" chapter in the Handbook.
Good luck!
Dave Hall -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
www.wormatlas.org www.aecom.yu.edu/wormem
phone 718 430-2195 fax 718 430-2514
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 10:48:23 2004
Dear Michael, When I need long-term SEM gun stability and long filament life, I use a well under-saturated filament. I don't think the first peak is stable enough. On my Hitachi instruments, I can use "filament image" mode to see an image of the filament tip, and back off the heating current until I have a nice doughnut with a dark centre. You can also use the waveform mode to determine the zero-emission level and the fully saturated level, then back off about 10 % of that range from the saturation point. This is above the exponential slope of filament brightness, when the saturation curve starts to curve over, and is fairly stable over days. 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 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Microscopy list" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, December 15, 2004 4:38 AM
Hello,
We are setting up a modest microscopy suite in our dept and are in need of a used glass knifemaker. Anyone know of a good source, or have a used one for purchase? You may respond to me personally, rather than the list.
Thanks, Heidi
Heidi A. Kratsch, Ph.D. Assistant Professor/Extension Ornamental Horticulture Specialist Utah State University Department of Plants, Soils & Biometeorology 4820 Old Main Hill Logan, UT 84322-4820 Phone: 435-797-8124 Mobile: 435-760-5611 Fax: 435-797-3376 Email: heidik-at-ext.usu.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 11:58:01 2004
What you are saying (to use the first peak) is new to me. I have a XL30 ESEM, Quanta's senior generation. Yes, there is a way to limit the filament current; but you have to adjust it from time to time. I am talking about the second peak here. I find that the auto saturation is usually over-done after I have had a few burned out prematurely. These days, I let the machine do auto saturation when it is new. I check it within 10 min to lower the LIMIT setting. The saturation point drops very quickly for the first few days, so I keep checking it to prolong it's life. I had one lasted for 6 months; mostly 2-3 months and the rest can be anywhere down to few days when I neglected to check the filament. I hope this helps.
Ann Fook Yang
-----Original Message----- } From: michael shaffer [mailto:michael-at-shaffer.net] Sent: Wednesday, December 15, 2004 7:39 AM To: Microscopy list
A SEM we are purchasing has an application that does not require optimum gun brightness, and for the sake of filament longevity (months of 24-7) runs the filament heat at less than saturation and on top of the 1st peak.
I have always tought that the phenomenon of the 1st peak must be a result of the Wehnelt's geometry as a function of heating and the zero-to-positive keV potential moving towards the filament tip ... that is, a phenomenon of note but not much more of a concern. Now, I find myself wanting of a better understanding for the sake of beam current stability ... i.e., operating on the first peak does not seem to be a very stable parameter for stable emission.
The SEM itself is a FEI Quanta, which apparently has a method of regulating the emission current. This must also play a role, and I haven't yet found a explanation of how this works.
Any discussion towards enlightenment much appreciated.
In the early 1970s I worked with the group from St Bartholomew's Hospital, London, to publish the first papers on crystal deposition disease. Look up Crocker, Leverson et al for help with the problems that you have. We worked on synovial fluid from the knee so you should find a good deal of help in this area.
Regards
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide www.emcourses.com tel +44 1280 816512 fax +44 1280 814007
----- Original Message ----- } From: "Allan Mitchell" {allan.mitchell-at-stonebow.otago.ac.nz} To: {microscopy-at-msa.microscopy.com} Sent: Wednesday, December 15, 2004 4:03 AM
Hi Michael
The 1950s conventional wisdom as to the setting of the filament was to move onto the plateau of the heating/signal level graph as the filament heating current was often unstable. This procedure overcame intensity changes should the filament heating current change.
As years have gone by and improvements have been made this "overkill", in more ways than one, has been corrected. Many manufacturers suggest running at first peak when ever possible and even the purest amongst us have run for a number of years in this way.
What about stability you may say? Well set a TEM on what would be first peak (maximum screen intensity when heating the filament) and watch the virtual source. I think you will become more likely to develop eye strain than the source will change in form; the stability is amazing!
I back this observation by my experiences with instruments that judge image brightness in the search for heavy elements in the mining industry. These automated instruments work 24 hours a day and must remain stable in order to have a constant probe current and hence a constant range of grey levels in order to carry out their tasks; the elemental composition is judged by grey level through BSE.
I would also add that running on first peak on a modern instrument offers remarkable results; 150,000X being not out of the question (15kV 10mm WD).
I have not and cannot explain first peak but I can say on a modern instrument it produces a very satisfactory source at } 10kV.
Regards Steve Chapman Protrain for Training and Consultancy in Electron Microscopy World Wide www.emcourses.com
-----Original Message----- } From: michael shaffer [mailto:michael-at-shaffer.net] Sent: 15 December 2004 12:39 To: Microscopy list
A SEM we are purchasing has an application that does not require optimum gun brightness, and for the sake of filament longevity (months of 24-7) runs the filament heat at less than saturation and on top of the 1st peak.
I have always tought that the phenomenon of the 1st peak must be a result of the Wehnelt's geometry as a function of heating and the zero-to-positive keV potential moving towards the filament tip ... that is, a phenomenon of note but not much more of a concern. Now, I find myself wanting of a better understanding for the sake of beam current stability ... i.e., operating on the first peak does not seem to be a very stable parameter for stable emission.
The SEM itself is a FEI Quanta, which apparently has a method of regulating the emission current. This must also play a role, and I haven't yet found a explanation of how this works.
Any discussion towards enlightenment much appreciated.
If you've got enough brightness to spare, why not run it in the valley between the 1st peak and the 'plateau'?
Better stability than the 1st peak, in my JEOL 840 experience (think of the mechanical analogue), and certainly longer filament life thjan the 'plateau', but I think both the 1st peak and the valley give a slightly larger spot than the 'plateau'.
cheers
rtch
} From: "michael shaffer" {michael-at-shaffer.net} To: "Microscopy list" {Microscopy-at-MSA.Microscopy.Com}
} Hello Listers: } } I have two multi-packs boxes (250 sheets per pack) of the small sized 6.5 } x 9 cm. 4489 film. It has an expiration of 12/2004 but has been stored in } a cool dry place. Anyone out there interested in it? Normally it sells } in the catalogs for $ 97.00 for 100 sheets. I would sell the 250 boxes } for $50.00 each plus shipping costs. Anyone need film out there? } } Karen } } Karen L. Bentley, M.S.(previously Jensen) } Associate Scientist & Project Manager } Electron Microscope Research Core } University of Rochester Medical Center } Rochester, NY 14642 } 585-275-1954 }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 14:12:56 2004
} On our older ISI, the signal is greatly increased when you go } from the first peak to the edge of the plateau (typical } saturation point). Even if adequate image brightness is achieved } on some SEMs at the first peak, am I wrong to think that more } signal means better S:N ratio, and therefore a better quality } image? And am I wrong to think that the only advantage to heating } the filament only to the first peak is increased filament life } (but at the expense of image quality)? Or are these more modern } SEMs very different from the older models?
I believe you are correct on both counts ... leastwise, I am not yet convinced what Steve implies below ... i.e., that emission from the cathode at 1st peak is equally bright as when saturated. Regarding signal/noise, do not confuse "spot brightness" with "beam current". For sure, increasing either results in better s/n ... but brightness is a function of your gun emission, and once your gun is properly saturated (whatever method), it is your beam current setting (e.g., spot size) which, in practice, controls your s/n. (except of course, your choice of final aperture)
cheerios :o)
} Steve Chapman 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 Michael } } The 1950s conventional wisdom as to the setting of the filament was to } move onto the plateau of the heating/signal level graph as the filament } heating current was often unstable. This procedure overcame intensity } changes should the filament heating current change. } } As years have gone by and improvements have been made this "overkill", } in more ways than one, has been corrected. Many manufacturers suggest } running at first peak when ever possible and even the purest amongst us } have run for a number of years in this way. } } What about stability you may say? Well set a TEM on what would be first } peak (maximum screen intensity when heating the filament) and watch the } virtual source. I think you will become more likely to develop eye } strain than the source will change in form; the stability is amazing! } } I back this observation by my experiences with instruments that judge } image brightness in the search for heavy elements in the mining } industry. These automated instruments work 24 hours a day and must } remain stable in order to have a constant probe current and hence a } constant range of grey levels in order to carry out their tasks; the } elemental composition is judged by grey level through BSE. } } I would also add that running on first peak on a modern instrument } offers remarkable results; 150,000X being not out of the question (15kV } 10mm WD). } } I have not and cannot explain first peak but I can say on a modern } instrument it produces a very satisfactory source at } 10kV. } } Regards } Steve Chapman } Protrain for Training and Consultancy in Electron Microscopy World Wide } www.emcourses.com } } } -----Original Message----- } } From: michael shaffer [mailto:michael-at-shaffer.net] } } Sent: 15 December 2004 12:39 } To: Microscopy list } Subject: [Microscopy] SEM: gun saturation } } } } ------------------------------------------------------------------------ } ------ } 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 } ------------------------------------------------------------------------ } ------- } } } A SEM we are purchasing has an application that does not require optimum } gun } brightness, and for the sake of filament longevity (months of 24-7) runs } the } filament heat at less than saturation and on top of the 1st peak. } } I have always tought that the phenomenon of the 1st peak must be a } result of } the Wehnelt's geometry as a function of heating and the zero-to-positive } keV } potential moving towards the filament tip ... that is, a phenomenon of } note } but not much more of a concern. Now, I find myself wanting of a better } understanding for the sake of beam current stability ... i.e., operating } on } the first peak does not seem to be a very stable parameter for stable } emission. } } The SEM itself is a FEI Quanta, which apparently has a method of } regulating } the emission current. This must also play a role, and I haven't yet } found a } explanation of how this works. } } Any discussion towards enlightenment much appreciated. } } Happy Holidays & cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } {www.micro-investigations.com} } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 14:20:20 2004
Hello experts. I have an application where I need a very hard resin formulation. I had been using Mollenhauer Araldite-Epon mixture using BDMA as the accelerator. This formulation is not hard enough. I am working with zebrafish which needs a prolonged infiltration schedule. I cannot use Spurr as I am very allergic to it. I tried the embed-it resin (a spurr like low viscosity mixture) with unpredictable results. Can anyone recommend a hard formulation that infiltrates tissue well using either Epon or Araldite? I plan to try the distilled DDSA that should infiltrate well. Thanks for your suggestions. JoAnn Buchanan
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 21:48:32 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mourad.idir-at-synchrotron-soleil.fr) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, December 15, 2004 at 14:14:30 ---------------------------------------------------------------------------
Question: Dear All I had the following questions 1-- Suppose we used A system with a CCD camera with 10 microns pixels size (1000x1000 pixels) and Microscope objective: "10X" with 0.1 NA This means that we will have a field of view of 1 x 1 mm and an effective pixel size of 1 micron. NA= 0.2 give a diffraction limited resolution of about 0.61x0.55/0.1=3.355 microns. If I understand correctly there is no need to have an objective with a x20 but it is better to have an objective with a x10 and higher NA. If we do the reverse calculation, we nedd two pixels for the resolution (Nyquist theorem) : 2 x10 microns pixel / (x10)=2 microns effective in the object plan so we need a NA of about NA=0.61x 0.5 / 2 = 0.168. With this I do not need to get a higher magnification because it is the pixel camera who limit the resolution of the image IS THAT CORRECT?
2-- The Depth of Field in an optical system is lambda/NA^2 = 0.5/(0.2)^2 = 12.50 microns. So if I have a sample who is emmiting light in 10 microns thickness this will limit the resolution in my image. This means that the image resolution in our case is limited to this Depth of Field so around 10 µm.
In other word, if we have a sample thickness of 10 µm this will limit the resolution but if we have a sample thickness equal to 2 microns we will have 2 microns resolution IS THAT CORRECT? Thanks for you reply Regards
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kalyani-at-jncasr.ac.in) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, December 15, 2004 at 09:11:03 ---------------------------------------------------------------------------
Email: kalyani-at-jncasr.ac.in Name: kalyani
Organization: JNCASR
Education: Graduate College
Location: Bangalore, Karnataka
Question: Sir,
} From an HREM image of a nanowire i am getting a d-spacing which shows up neither in the SAED pattern nor in the XRD pattern. even other HREM images of nanowires of the same sample also show that d-spacing itself. but in the XRD pattern, a peak with half of the d-spacing that is calculated from the HREM image,is present. is there any particular reason for this observation. if yes, could u please give some egs as well as references.
} } Jan writes ... } } } On our older ISI, the signal is greatly increased when you } go from the } } first peak to the edge of the plateau (typical saturation } point). Even } } if adequate image brightness is achieved on some SEMs at the first } } peak, am I wrong to think that more signal means better S:N } ratio, and } } therefore a better quality image? And am I wrong to think that the } } only advantage to heating the filament only to the first peak is } } increased filament life (but at the expense of image } quality)? Or are } } these more modern SEMs very different from the older models? } } I believe you are correct on both counts ... leastwise, I } am not yet convinced what Steve implies below ... i.e., that } emission from the cathode at 1st peak is equally bright as } when saturated. Regarding signal/noise, do not confuse "spot } brightness" with "beam current". For sure, increasing either } results in better s/n ... but brightness is a function of } your gun emission, and once your gun is properly saturated } (whatever method), it is your beam current setting (e.g., } spot size) which, in practice, controls your s/n. (except of } course, your choice of final aperture)
I believe the quality of the signal and stability from the first peak depend very much on instrument in use. But I have not found a single microscope stable enough on the first peak for prolonged EDS or WDS analysis. So, while image quality could be satisfactory with the gun at the first peak, for proper microanalysis gun saturation is a necessity.
} cheerios :o) } } } Steve Chapman 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 Michael } } } } The 1950s conventional wisdom as to the setting of the } filament was to } } move onto the plateau of the heating/signal level graph as the } } filament heating current was often unstable. This } procedure overcame } } intensity changes should the filament heating current change. } } } } As years have gone by and improvements have been made this } "overkill", } } in more ways than one, has been corrected. Many } manufacturers suggest } } running at first peak when ever possible and even the } purest amongst } } us have run for a number of years in this way. } } } } What about stability you may say? Well set a TEM on what would be } } first peak (maximum screen intensity when heating the filament) and } } watch the virtual source. I think you will become more likely to } } develop eye strain than the source will change in form; the } stability } } is amazing! } } } } I back this observation by my experiences with instruments } that judge } } image brightness in the search for heavy elements in the mining } } industry. These automated instruments work 24 hours a day and must } } remain stable in order to have a constant probe current and hence a } } constant range of grey levels in order to carry out their } tasks; the } } elemental composition is judged by grey level through BSE. } } } } I would also add that running on first peak on a modern instrument } } offers remarkable results; 150,000X being not out of the question } } (15kV 10mm WD). } } } } I have not and cannot explain first peak but I can say on a modern } } instrument it produces a very satisfactory source at } 10kV. } } } } Regards } } Steve Chapman } } Protrain for Training and Consultancy in Electron Microscopy World } } Wide www.emcourses.com } } } } } } -----Original Message----- } } } } From: michael shaffer [mailto:michael-at-shaffer.net] } } } } Sent: 15 December 2004 12:39 } } To: Microscopy list } } Subject: [Microscopy] SEM: gun saturation } } } } } } } } } ---------------------------------------------------------------------- } } -- } } ------ } } 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 } } } -------------------------------------------------------------- } ---------- } } ------- } } } } } } A SEM we are purchasing has an application that does not require } } optimum gun brightness, and for the sake of filament } longevity (months } } of 24-7) runs the } } filament heat at less than saturation and on top of the 1st peak. } } } } I have always tought that the phenomenon of the 1st peak must be a } } result of the Wehnelt's geometry as a function of heating and the } } zero-to-positive keV } } potential moving towards the filament tip ... that is, a } phenomenon of } } note } } but not much more of a concern. Now, I find myself wanting } of a better } } understanding for the sake of beam current stability ... } i.e., operating } } on } } the first peak does not seem to be a very stable parameter } for stable } } emission. } } } } The SEM itself is a FEI Quanta, which apparently has a method of } } regulating the emission current. This must also play a role, and I } } haven't yet found a } } explanation of how this works. } } } } Any discussion towards enlightenment much appreciated. } } } } Happy Holidays & cheerios ... shAf :o) } } Avalon Peninsula, Newfoundland } } {www.micro-investigations.com} } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 16 09:07:07 2004
Araldite tends to make a softer resin. For hard tissue, I use:
Epon 40g DDSA 10g NMA 20g
Infiltrate with this mix for some time with several changes, then add DMP30 2% to the pre-mixed Epon, mix even more thoroughly and infiltrate for a further period before embedding in fresh Epon + DMP30 mix. The NMA (nadic methyl anhydride) makes the polymerised resin much harder than normal Epon.
Lesley Weston.
on 15/12/2004 12:50 PM, JoAnn Buchanan at redhair-at-stanford.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 } ---------------------------------------------------------------------------- --} - } } Hello experts. I have an application where I need a very hard resin } formulation. I had been using Mollenhauer Araldite-Epon mixture using } BDMA as the accelerator. This formulation is not hard enough. } I am working with zebrafish which needs a prolonged infiltration schedule. } I cannot use Spurr as I am very allergic to it. I tried the embed-it resin } (a spurr like low viscosity mixture) with unpredictable results. } Can anyone recommend a hard formulation that infiltrates tissue well using } either Epon or Araldite? I plan to try the distilled DDSA that should } infiltrate well. } Thanks for your suggestions. } JoAnn Buchanan } } Department of Molecular and Cellular Physiology } Stanford University School of Medicine } Stanford, CA 94305 } 650-723-5856 } }
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 16 09:20:41 2004
I have some nice pictures taken on the first peak where one can see the same objets twice or three times a bit shifted one from the other. Setting the filament current to saturation, one see that one image dominate, while the others are disapering.
The explanation is that there are some hot points on the filament, hot enough to emit, and so there are two or three emission points. With more current the filament tip become more homogeneous in temperature, and at saturation there is only the tip which is the emitting point (in combination of the wehnelt polarisation). With time, evaporation and annealing of the W make that the surface of the filament become smoother, and so these points will desapear.
The ability to work on the first peak or on the plateau depends more of the quality of the W wire used to make the filament, and of the annealing effect of a few hours of work, than of the age/generation of the microscope. It's the same question thant the lifetime of the filaments. Depends on how the microsope is driven, but on the filament quality too.
On our Auger e-guns, we never work at saturation, and filament hold 10 to 15 years. But there is no image, and the shape of the electron spot does'nt matter. We need only electrons at à given energy and current.
On the other hand, if the first peak is economically interesting, its shape, position an intensity often change during the life of the filament. Sometimes it's very difficult to see it, sometimes it has a very strong intensity... For a beginner, it can give headache to find it. Saturation is easier. Over saturation too !
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
according to manufacturer's instruction (SERVA, leaflet dated 1997), the very hard grade Glycid ether 100 resin (equivatent to the former Epon 812) is obtained as follows:
52.9 ml of glycid ether 47.1 ml MNA mix thoroughly, then add 1.5 ml DMP-30 (or 2,4,6 trisphenol...), again mix thoroughly
polymerisation: 20 h at 45oC, then 24 h at 60oC
I wish you success :-) Barbara Lotocka
dr Barbara £otocka Department of Botany Faculty of Agriculture and Biology Warsaw Agricultural University Nowoursynowska 159 02-776 Warsaw, Poland
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 16 11:58:11 2004
What type of info are you looking for from your image?
Last week at Cell Bio, I had a chance to see the new integration of NTMDT's atomic force microscope with a microtome. Dr. Anton Efimov, the inventor, had prepared a nematode using the normal freeze substitution protocols for TEM and then had done a serial section and 3D reconstruction, using a diamond knife. Because the knife cuts so cleanly, there is no topography info to be gathered. However, the AFM has the capability to image using local elasticity. The results were intriguing.
If you are interested in further info, I'd suggest you correspond directly with Dr. Efimov (efimov-at-ntmdt.ru).
Hope this was helpful,
Best regards, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
Caveat: MME has financial interest in this product. At 09:36 AM 12/15/2004, Renaat Dasseville wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Dec 16 15:58:40 2004
For others who may have spectrometer interfaces which use the old ISA slots, but wish to upgrade their computers, Soltek now makes a motherboard (SL-XP865G- 3IG) which uses the Intel 865G chipset, has FSB of 400, 533, or 800MHz, supports Pentium 4 CPU, and has 4 PCI and 3 ISA slots.
It also has integrated graphics, a LAN port, and 4 integrated USB 2.0 ports, but no built-in sound.
I have no connection with Soltek or its agents, except as a satisfied customer.
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 Thu Dec 16 17:47:27 2004
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From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 04:27:48 2004
} [...] } I thought the point of the original question was to determine } our experiences of using the desaturated filaments and the } stabilities involved? } In which case the experiences with these automated instruments } running desaturated would confirm a high stability over a period } of up to 24 hours. } [...]
My original question was for better understanding the phenomenon of the 1st peak. What's going on with electron emission with changing equipotential voltage influences? Why don't we see the monotonic rise as described by "self-biasing", but instead hills and valleys? I was hoping someone had written it up as something other than a curiosity. For example, modifying the cathode-Wehnelt distance may have some influence on the 1st peal, possibly making it a more stable point of heating.
I think most would agree ... putting filament heat on top of the 1st peak is not the most stable situation for long term electron emission. The instruments I am just now becoming more familiar with employ the ability to measure the beam current with regularity, and to digitally optimize the heat (find the top of the 1st peak), and control the first lens for making the beam current what it should be. These algorithms takes only a few seconds, but allow for the gun being almost maintenance-free while emitting equivelent current. This is somewhat different than relying on the inherent stability of the emission plateau (saturation) ... and also different from the method employed by microprobes which compensate with the first lens only.
I should now remain quiet until I have actually had my hands on with these SEMs. Still, when I'll need manage and teach these instruments, I'll be wanting for a better explanation of the "1st peak".
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (thokar7-at-in.gr) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, December 17, 2004 at 03:47:14 ---------------------------------------------------------------------------
I was just browsing through my newly arrived Nov 2004 copy of Microscopy Today and found something that has deeply shaken my confidence in my instrument and ability to operate it. The back inside cover (an ad for an SEM manufacturer who shall remain nameless) displays a very colorful FE-SEM image of Paralia sulcata (Phytoplankton), measuring some hundreds of microns across and looking nothing like the specimens of the same species I routinely image with my lowly tungsten-based instrument (see http://www.mta.ca/dmf/wii23.htm for what I'm used to seeing).
It would appear that my SEM is generating not only a thoroughly spurious image, but also my magnification calibration is off by a factor of something like 300.
I understand that image quality is a function of the cost of the instrument, but the differences here are much more extreme than I ever appreciated. Why, it's like the two instruments are not even examining specimens for the same Kingdom! I guess my problem also stems from the fact that I can only rely on the 30+ years of expertise my wife (and collaborator) has in phytoplankton research, compared to the research budget of a heavy industry that is larger than the National Science Foundation. Obviously, I need to invest in better instrumentation. If one of the vendors will agree to fly me to Hawaii for the MSA meeting this summer, I'll almost certainly buy a new FE-SEM from them!
{tongue-in-cheedk mode OFF}
Wishing everyone a Happy (and Accurate) Holiday season,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Dear Listers, Our facility has an Ilford print processor which is not working. Parts are no longer available and so I'm looking for a replacement. The majority of our users use the digital camera but we have a few holdouts who want film. Can any of you recommend a black and white processor?
Thanks,
Mary Gail Engle Laboratory Manager Electron Microscopy & Imaging Facility University of Kentucky Lexington, KY 40536
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 09:58:32 2004
i am no whale expert but the humpback whale in the accompanying photo is also a tad larger than I would have guessed. The flukes look to span over 20 meters...
At 08:49 AM 12/17/04, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
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When Jerry Sedgewick's articles first appeared in Microscopy Today, I wrote a lengthy correction to some of his misinformation and sent it to Ron Anderson. There was no reply, and nothing appeared in the magazine, but Ron later told me he had forwarded the letter to Jerry, who has never responded. Realizing that the magazine is not a referreed journal and probably isn't taken very seriously anyway, I put the continuing errors in the series of artices in the category of a minor annoyance.
But in the most recent issue, at the end of Jerry's article, he has inserted the statement 'Note: More thorough explanations of the concepts presented in this article can be found in "The Image Processing Handbook" by John Russ.' By invoking my name and book in an apparent attempt to gain some credibility for the article, which is almost entirely incorrect and misleading, that statement requires an answer.
First, the cited book does not discuss the subject of matching the appearance of images on printouts and the computer screen. I have written in another book on that topic, but the bible on the subject can be found in David Blatner and Bruce Fraser's "Real World Photoshop" (Peachpit Press). Another excellent choice for anyone concerned about doing this right is to get either the GretagMacbeth Eye-One (my preference) or ColorVision Spyder calibration hardware and software, and follow the straightforward procedures which will make sure that your display, printout, and even projector all show the same thing.
The Sedgewick article is completely wrong (as usual). The International Color Consortium begun by companies like HP and Apple has constructed a technically sound and easily implemented framework that allows manufacturers of software (e.g., Photoshop), hardware (display cards, monitors, printers) and consumables (ink, paper) to provide widely available curves that everyone should use. Bypassing this (per Jerry's recommendation) and attempting to adjust the image or the monitor to make images more dramatic is a bad idea. Altering the contrast of "scientific" images as compared to normal photography is nonsense. Ink jet printers are fine and have their place, but are certainly not the only way to get good output. The article has plenty of other, more subtle errors (e.g., the dialog shows that sRGB color space is being used, instead of one such as Adobe 1998 with its greater gamut).
Ignore the article. In fact, ignore all of the Sedgewick articles about Photoshop. Even in those cases where the instructions are not wrong, they are not the best way to accomplish a given result. Go read a good book. There are lots of them, written by people who actually know what they are writing about.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 11:18:29 2004
You've let the cat out of the bag. That's actually a secret image of a new, planktonic mutant of Arabidopsis, showing a trichome engineered to increase the plant's buoyancy.
Phil
} Hi Listers, } } {tongue-in-cheek mode ON} } } I was just browsing through my newly arrived Nov 2004 copy of Microscopy Today } and found something that has deeply shaken my confidence in my instrument and } ability to operate it. The back inside cover (an ad for an SEM } manufacturer who } shall remain nameless) displays a very colorful FE-SEM image of } Paralia sulcata (Phytoplankton), } measuring some hundreds of microns across and looking nothing like } the specimens of the same } species I routinely image with my lowly tungsten-based instrument } (see http://www.mta.ca/dmf/wii23.htm for what I'm used to seeing). } } It would appear that my SEM is generating not only a thoroughly } spurious image, but also my } magnification calibration is off by a factor of something like 300. } } I understand that image quality is a function of the cost of the } instrument, but the differences } here are much more extreme than I ever appreciated. Why, it's like } the two instruments } are not even examining specimens for the same Kingdom! I guess my problem also } stems from the fact that I can only rely on the 30+ years of } expertise my wife (and collaborator) } has in phytoplankton research, compared to the research budget of a } heavy industry that is } larger than the National Science Foundation. Obviously, I need to invest } in better instrumentation. If one of the vendors will agree to fly } me to Hawaii for the } MSA meeting this summer, I'll almost certainly buy a new FE-SEM from them! } } {tongue-in-cheedk mode OFF} } } Wishing everyone a Happy (and Accurate) Holiday season, } } Jim } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/dmf
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 11:26:00 2004
[This is getting a bit off-topic of W vs. FE SEM, but...] I had similar thoughts first time I saw that image. I'll stick my neck out and suggest that its really a leaf of Arabadopsis, wild type. At least that's the consensus around here. Any other guesses??
Gib
-- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra
} Hi Listers, } } {tongue-in-cheek mode ON} } } I was just browsing through my newly arrived Nov 2004 copy of Microscopy } Today } and found something that has deeply shaken my confidence in my } instrument and } ability to operate it. The back inside cover (an ad for an SEM } manufacturer who } shall remain nameless) displays a very colorful FE-SEM image of Paralia } sulcata (Phytoplankton), } measuring some hundreds of microns across and looking nothing like the } specimens of the same } species I routinely image with my lowly tungsten-based instrument } (see http://www.mta.ca/dmf/wii23.htm for what I'm used to seeing). } } It would appear that my SEM is generating not only a thoroughly spurious } image, but also my } magnification calibration is off by a factor of something like 300. } } I understand that image quality is a function of the cost of the } instrument, but the differences } here are much more extreme than I ever appreciated. Why, it's like the } two instruments } are not even examining specimens for the same Kingdom! I guess my } problem also } stems from the fact that I can only rely on the 30+ years of expertise } my wife (and collaborator) } has in phytoplankton research, compared to the research budget of a } heavy industry that is } larger than the National Science Foundation. Obviously, I need to invest } in better instrumentation. If one of the vendors will agree to fly me to } Hawaii for the } MSA meeting this summer, I'll almost certainly buy a new FE-SEM from them! } } {tongue-in-cheedk mode OFF} } } Wishing everyone a Happy (and Accurate) Holiday season, } } Jim
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 11:46:59 2004
Looking for labs that have access to LAICP mass spec or NAA to look at trace rare earth elements in apatite and tourmaline mineral samples. Need quantification so not sure if you folks in the SIMS area can help but thought I would ask. If you have any ideas please respond directly with contact info, resource to be used, and poor university pricing. Will consider commercial labs.
Thanks
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 13:34:17 2004
EMS now does this kind of things - see http://www.emsdiasum.com/default.htm
At 12:48 PM 12/17/2004, Karl Hagglund wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 13:47:37 2004
I should offer at least one tale of warning in connection with ISA cards.
We had an EDS computer for one of our SEMs. The EDS system (Oxford ISIS) required 1 ISA slot for its interface and we had an older PCI image capture system on the same computer and that required two ISA slots. We upgraded the motherboard over the years from a 486-50 to a Pentium 90, to a Pentium 200 Overdrive processor, to an AMD 500. That all went well as we always ordered motherboards with 3 (or more) ISA slots.
When it came time to make the next jump, we could not find a motherboard with three slots. I found a Biostar board with 1 slot. We bit the bullet and spent the money to upgrade the PCI system to a single PCI card and tried to keep the ISIS running on the ISA card. That did NOT work.
We ran into a variety of problems, probably related to timing issues. We had not changed any software except a few drivers for the motherboard. We could switch back to the old motherboard and things would work fine, but swap in the new motherboard and things got weird again. Consultation with the staff at Oxford confirmed the timing suspicion. We determined the best course would be to upgrade the ISIS board from ISA to PCI.
However, even that did not work completely smoothly. We had upgraded from Windows 3.11 through Windows 95 to Windows 98se over the years with no problem. The driver needed for the new PCI card only worked properly under Windows 2000 or XP. Upgrading to 2000 solved the problem and our system has been running fine for the last two years.
I may add that we tried to upgrade to Windows XP, but that led to other problems. XP does not work with one an image library (KNIFE.VBX) that is required for one piece of our software.
The moral of the story seems to be that upgrades are not always as painless as we would like them to be. While we were able to change out motherboards and upgrade Windows several times, there can be problems waiting at the next upgrade. I am happy to see the availability of motherboard that again includes ISA slots, but I would recommend caution when trying to upgrade with it. The specialized cards that we use are not so common and not so universally tested as a sound or network card. Problems could be lurking.
Disclaimer: The folks at Oxford were reasonably helpful and pleasant throughout this process. I can imagine the anxiety on the part of a tech support person when a user says they are about to embark on a upgrade. The process is not under the control of tech support and a lot of things that can (and do) go wrong. Oxford has been fair to point out that they could not guarantee the outcome of any of our upgrades, but they have helped us as they could.
Warren
At 04:28 PM 12/16/04, you wrote:
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------------------------------------------- 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
Fred Schamber wrote a series of articles in Microscopy Today (Sep - Nov 1999) on triode electron guns. I scanned them in and use it for my class (with permission). You can find it at
In my discussions with Fred, we talked a little about the first ("false") peak. The first peak does not show up in all guns and seems to be a function of the gun design. It seems to more prevalent in older designs than in the newer guns. Fred indicated that he did not see it in the Personal SEM (TM) gun he designed. Our speculation at the time was that the false peak could be due to emission from the sides of the W loop filament. This was based on my experience with looking at TEM filament images.
If you talk with Fred, he will discuss the equipotential lines as the wehnelt bias is varied. I usually visualize the wehnelt as an electrostatic lens. As the potential (bias) is increased, the focal length is shortened. Saturation occurs when the focal length is such that you are looking at just the tip of the filament. At the lower bias voltages, you tend to see the halo around the central image of the tip. These seem to be coming from the "side" faces of the wire loop. I suspect that these electrons are responsible for the false peak. However if Fred wishes to jump into this thread, I'll certainly defer to him!
Cheers, Henk
At 05:57 AM 12/17/04, michael shaffer wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 14:35:39 2004
First, let me say that I do not have any issues with the technical issues that you put in response to Jerry's article because I do not consider myself an expert and do consider you one. When you write an article or submit something on the listserver, my ears perk up and I read it. I have personally benefited from your generosity in discussing and helping me with stereological problems. However, there are a number of points that you raise that I think that should be addressed.
You are correct that MT is not a refereed journal, but I don't think that your assessment that people don't take it seriously is correct. Ron is an experienced microscopist and does a very good job of putting solid articles together in a coherent fashion. For the most part, the articles are by people that are well-respected. I have written a few articles for MT (not that I am well-respected, LOL) and I am careful to try not to put anything that is incorrect in there because I realize that it is not refereed. It also takes a bit of time to put an article together and I thank the efforts of those who have written for MT. I have talked to others who have written articles and for the most part, they have taken similar care. If you have technical exceptions to an article that someone has written, I would bet that Ron would be willing to include those if they were written in an objective mode.
Next, the Listserver is for discussion. Your points with respect to the MT article are valid and are on subject and definitely should generate discussion. However, I think that your delivery was a bit personal and rather scathing. That wasn't appropriate for the listserver and was against the rules. John, forgive me for saying this, but it looks unprofessional and lacked tact. It looked as if you were frustrated with nobody listening to you. Believe me that that is not the case when you talk about imaging. I just was not comfortable reading your submission because of the personal attack that it had.
I wrote this because I don't want people afraid to submit either to the listserver or MT for fear of personal attacks by well-known people in the field. I hope that this has not jeopardized our friendship in any way. These are my opinions alone.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472 Walck-at-PPG.com (412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Friday, December 17, 2004 12:20 PM To: Microscopy-at-msa.microscopy.com
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When Jerry Sedgewick's articles first appeared in Microscopy Today, I wrote a lengthy correction to some of his misinformation and sent it to Ron Anderson. There was no reply, and nothing appeared in the magazine, but Ron later told me he had forwarded the letter to Jerry, who has never responded. Realizing that the magazine is not a referreed journal and probably isn't taken very seriously anyway, I put the continuing errors in the series of artices in the category of a minor annoyance.
But in the most recent issue, at the end of Jerry's article, he has inserted the statement 'Note: More thorough explanations of the concepts presented in this article can be found in "The Image Processing Handbook" by John Russ.' By invoking my name and book in an apparent attempt to gain some credibility for the article, which is almost entirely incorrect and misleading, that statement requires an answer.
First, the cited book does not discuss the subject of matching the appearance of images on printouts and the computer screen. I have written in another book on that topic, but the bible on the subject can be found in David Blatner and Bruce Fraser's "Real World Photoshop" (Peachpit Press). Another excellent choice for anyone concerned about doing this right is to get either the GretagMacbeth Eye-One (my preference) or ColorVision Spyder calibration hardware and software, and follow the straightforward procedures which will make sure that your display, printout, and even projector all show the same thing.
The Sedgewick article is completely wrong (as usual). The International Color Consortium begun by companies like HP and Apple has constructed a technically sound and easily implemented framework that allows manufacturers of software (e.g., Photoshop), hardware (display cards, monitors, printers) and consumables (ink, paper) to provide widely available curves that everyone should use. Bypassing this (per Jerry's recommendation) and attempting to adjust the image or the monitor to make images more dramatic is a bad idea. Altering the contrast of "scientific" images as compared to normal photography is nonsense. Ink jet printers are fine and have their place, but are certainly not the only way to get good output. The article has plenty of other, more subtle errors (e.g., the dialog shows that sRGB color space is being used, instead of one such as Adobe 1998 with its greater gamut).
Ignore the article. In fact, ignore all of the Sedgewick articles about Photoshop. Even in those cases where the instructions are not wrong, they are not the best way to accomplish a given result. Go read a good book. There are lots of them, written by people who actually know what they are writing about.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 15:41:52 2004
I have been relatively quiet about postings lately. But I posted today and received the normal "prize" for my offering - multiple out-of-the-office notices.
Since many of us will be taking time off in the next couple of weeks, please allow me to preempt Nestor and remind you all of the guidelines in the list FAQ. They are reproduced below. Please do not enable the out-of-office reply without unsubscribing from the list.
Are There Any Special Settings on my Email Program?
---------- Yes.
1.) Please insure that your correct Email address is listed in the FROM: record. You may need to enter this in the Preferences option of your Email program. Contact your system administrator if your not sure what to do here.
2.) DO NOT set your Email Program to automatically reply to all messages, or to request a return receipt. If either of these are done, you run the risk of creating an Email loop within the system. This may result in a message bouncing through the system for several days, filling up everyone's mail box!
3.) Generally it is best to terminate each line of your message at less than 80 characters. This avoids the occasional problem of messages getting chopped up. Some mail routers, limit the number of characters per line. If you exceed 80 characters it is sometimes possible that your message will become randomly truncated. If your not sure, then simply enter a (Carriage Return) at the end of each line of text you type (i.e., don't let your system automatically word wrap) and things should be fine.
4.) Make sure you do not send attachments of any type. (see above)
5.) Do not set your Email program to deliver "out of the office/vacation" messages. If you will be gone, you should unsubscribe from the list and subscribe again when you return.
6.) It is not out of line to provide your company name, Email address or WWW site as part of your signoff/signature line, at the end of ANY message you post to this system. Anonymous posting of Email is frowned upon.
------------------------------------------- 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
Scanning electron microscopy, x-ray analysis, and image analysis of materials Computer applications and networking ------------------------------------------- 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
I've responded to you privately about some of the points in your posting. For the public record, let me say that I do not mean to imply in any way that everything in the magazine is of poor quality - much of it is quite good. And I've written articles for it myself. But the quality depends on the efforts of the submitter, not on any review process, so readers need to exercise some judgment.
My posting was not intended as a personal attack on either Jerry or Ron. Ron has to fill the magazine for each issue and must depend on whatever people are willing to send him. The simple fact is that the Sedgewick articles on Photoshop (and his book put out by Plenum) contain many, many errors, both on image processing in general and Photoshop in particular. I don't intend to try to correct them point by point. There are plenty of texts on both image processing and on Photoshop that get it right, and people should be warned to read them.
I have not responded publicly in the past to the errors in the articles, but this time the fact that my name and book were referenced inappropriately and incorrectly in the article required some kind of corrective response. Sorry if I offended you....
John Russ
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 22:02:10 2004
Karl Hagglund wrote: ================================================================ } Hello, } } We have a See Vac Critical Point Dryer in need of repair. Does anyone } know of a third party company that performs repairs on these devices. ================================================================ The See Vac firm went out of existence around 1983 or 1984 (if I remember correctly). The owner and president, Blair Seese, passed away about ten or twelve years ago from cancer.
We ended up with a box of spare parts, mainly for the Seevac manufacturered sputter coaters and also some of the other equipment items they made, including the CPD unit. The CPD unit has not been made since the early 1980's. If you could contact me off-line and tell me what parts you think you need, I could see if we have them.
The system you have is roughly 25 years old. You might want to talk to your safety people and get their opinion on the need to get it safety re-tested. There are special laboratories who do that sort of thing, we are not one of them.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 22:28:43 2004
I am a beginner in using TEM. I am currently in investigating nano-particles of 20nm and below. However, I have difficulty in getting the diffraction pattern in TEM. May I know if i can have some advice on this issue? Thanks a lot!
Regards, Yee Yan Postgraduate Nanyang Technological University Singapore
__________________________________________________ Do You Yahoo!? Download the latest ringtones, games, and more! http://sg.mobile.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 18 07:25:42 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mgrace-at-fit.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html#form on Friday, December 17, 2004 at 12:40:39 ---------------------------------------------------------------------------
Email: mgrace-at-fit.edu Name: Michael Grace
Organization: Florida Institute of Technology
Education: Graduate College
Location: Melbourne, FL USA
Question: I am purchasing a digital imaging system for a film-based TEM (Zeiss EM900), and am trying to decide on a camera. Questions:
1. I am currently considering the SIS MegaView and Morada, and the AMT XR40/XR60. Does anyone have experience with these? Recommendations?
2. One ultimate goal is to have high quality digital images for publication. How much resolution in a system is enough? Pixel size/number info is difficult (for me, at least) to understand and to relate to actual output resolution.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zohrehhamnabard-at-yahoo.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, December 18, 2004 at 01:38:06 ---------------------------------------------------------------------------
Question: dear sir/madam, I have some glass-ceramic samples(phlogopite system) and I am going to investigate them through SEM. How can I etch these samples and what etchants can I use for this purpose to get best results and high degree photos.
I am looking for FIB services for TEM samples (universities or companies). I prefer a training course of FIB where I can bring my own samples, depending on the cost. If there are such services (training, sample preparation services, or just free testing my samples), please advise.
Thank you, Hiromi Konishi, Ph.D.
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 03:23:34 2004
Dear Yee-Yan, This is not exactly a beginner's problem and is in fact quite hard. If you want powder diffraction patterns, then put a large SA aperture in and hit Diffraction, easy enough. If you want single particle diffraction patterns, then SA will probably not give sufficient intensity from such small particles. You will probably have to use some form of convergent beam diffraction. Whether this is done in conventional imaging mode, or using a nanodiffraction mode is up to you and your microscope. Whatever, you need a focussed probe with small spot size, sufficient intensity to see a diffraction pattern without roasting the particle, and a convergence angle low enough to separate the diffraction spots (may need to change the C2 aperture). Then press diffraction. This is fiddly but should work. The really tricky bit is if you then need to tilt the particle to a specific orientation. I can only say, be patient! And don't expect anything nice like Kikuchi bands to help you, it is too thin.
Best wishes
Ian
} I am a beginner in using TEM. I am currently in } investigating } nano-particles of 20nm and below. However, I have } difficulty in getting the } diffraction pattern in TEM. May I know if i can have } some advice on this } issue? Thanks a lot! }
-- Ian MacLaren Department of Physics and Astronomy University of Glasgow Glasgow G12 8QQ Scotland http://www.ssp.gla.ac.uk/
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 08:04:05 2004
Did anyone ask if Yee-Yan's particles are crystalline? Properly recording and interpreting the results aside, E.D. is simple enough that anyone with an instruction book or a colleague nearby can work the controls of a TEM to get some sort of a diffraction pattern on the screen.
Yee-Yan: if you illuminated some of your particles with an electron beam in a TEM set in diffraction mode, and all you see is a single bright spot centered on a featureless, diffuse background, it could be that your particles are amorphous (or nearly so).
Ron Anderson
-----Original Message----- } From: Ian MacLaren [mailto:i.maclaren-at-physics.gla.ac.uk] Sent: Monday, December 20, 2004 4:22 AM To: Tay Yee Yan Cc: Microscopy-at-msa.microscopy.com
Dear Yee-Yan, This is not exactly a beginner's problem and is in fact quite hard. If you want powder diffraction patterns, then put a large SA aperture in and hit Diffraction, easy enough. If you want single particle diffraction patterns, then SA will probably not give sufficient intensity from such small particles. You will probably have to use some form of convergent beam diffraction. Whether this is done in conventional imaging mode, or using a nanodiffraction mode is up to you and your microscope. Whatever, you need a focussed probe with small spot size, sufficient intensity to see a diffraction pattern without roasting the particle, and a convergence angle low enough to separate the diffraction spots (may need to change the C2 aperture). Then press diffraction. This is fiddly but should work. The really tricky bit is if you then need to tilt the particle to a specific orientation. I can only say, be patient! And don't expect anything nice like Kikuchi bands to help you, it is too thin.
Best wishes
Ian
} I am a beginner in using TEM. I am currently in } investigating } nano-particles of 20nm and below. However, I have } difficulty in getting the } diffraction pattern in TEM. May I know if i can have } some advice on this } issue? Thanks a lot! }
-- Ian MacLaren Department of Physics and Astronomy University of Glasgow Glasgow G12 8QQ Scotland http://www.ssp.gla.ac.uk/
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 08:24:03 2004
We have a LA-ICP-MS, but not for commercial use. Two companies that you can check out include Balazs (there is a local office in the DFW area, but the lab is in California) and Shiva in New York, but they do Glow Discharge MS. Both are pretty good companies (we still do business with both companies)...with one strong caveat. Both companies apparently believe that their trace analysis capability with the instrumentation cited does not need need a matrix-matched standard or reference material. Their explanations for no need of reference materials are, I believe, nothing but smoke and mirrors. In order to get results that are dependable, a standard that is a well characterized apatite or tourmaline...or close analogue is needed. We don't believe one can calibrate either instrument with, say, a NIST glass reference, and then expect to get good results analyzing SiC.
Contact me off-line if you wish.
Chuck Butterick Analytical Laboratory Supervisor Poco Graphite, Inc. 300 Old Greenwood Road Decatur, TX 76234 (940) 393-4287 (Phone) (940) 393-8383 (Fax) CButterick-at-poco.com
-----Original Message----- } From: Beavers, Roy [mailto:rbeavers-at-mail.smu.edu] Sent: Friday, December 17, 2004 12:17 PM To: microscopy-at-microscopy.com
Group,
Looking for labs that have access to LAICP mass spec or NAA to look at trace rare earth elements in apatite and tourmaline mineral samples. Need quantification so not sure if you folks in the SIMS area can help but thought I would ask. If you have any ideas please respond directly with contact info, resource to be used, and poor university pricing. Will consider commercial labs.
Thanks
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 11:50:22 2004
} I am a beginner in using TEM. I am currently in } investigating } nano-particles of 20nm and below. However, I have } difficulty in getting the } diffraction pattern in TEM. May I know if i can have } some advice on this } issue? Thanks a lot! } Dear Yee Yan, One problem you might have is that such small particles make up a very small fraction of the area seen in even the smallest selected area aperture you have in your scope. Unless the particles constitute a reasonable fraction of the scattering, the pattern will be lost in the power spectrum of the substrate in the selected area. My advice is to use the thinnest possible support film, make that film from low-Z material (carbon is probably best), increase the concentration of nanoparticles in your prep, and put a very small aperture (5 or 10 um) in the SA aperture holder and use it to restrict the area. You will probably see only diffraction rings, since the hoped-for many particles in the selected area will not, in general, be oriented in the same direction. 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 Dec 20 13:54:32 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You also need to keep in mind that with SA diffraction, spherical aberation limits how precisely you can define the area from which you get diffraction. The error is of the order of 500 nm.
Typically, the smallest practical SA aperture will be ~5 um which results in a selected area of ~ {500 nm. Consequently, if you really push SA diffraction and succeed in getting a pattern, it is likely to be from a region different from where you think it is coming from.
Best regards, -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 13:59:48 2004
The 4489 film I mentioned last week has been sold to the first responder to my email. Thanks for all of your inquiries!
Happy Holidays to everyone who replied!
Karen Bentley
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 17:18:52 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kjl226-at-vt.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, December 20, 2004 at 15:05:08 ---------------------------------------------------------------------------
Question: Has anyone out there had any expereince working with the Zeiss EVO40 SEM? I would like to get opinions on what you think of the instrument. Our Department purchased a EVO40 about 6 months ago and have had several serious problems. We are wondering if we have purchased a lemon.
======================================= First Notice of Microscopy Listserver DataBase Update: ======================================= Dec. 21, 2004
Colleagues....
After nearly a dozen years of operation, the master database of the Microscopy Listserver needs a serious reworking and purging. I have done the hard work of restructuring the software model but now the database needs to be "repopulated" with updated/new user information.
The easiest way for me to accomplish this is to simply purge the entire database, and have each of you resubscribe as if you are a new user.
As a result, I'm asking EACH and EVERY ONE of you to visit the following WWW site and fill out the new subscription form. Here is the URL:
You can do this beginning immediately, the changes will be stored and I will implement the new system on or about the second week of Jan. 2005. If all goes well, resubscribing should only take a few minutes of your time, so please do it as soon as possible.
For the balance of 2004 the old database will remain in use, but if you have not resubscribed by ~ the first week of Jan 2005, you will no longer receive Listserver Email as I will cease using the old database and only forward Listserver Email to those individuals who have subscribed and are recorded in the new system.
I'll post this message every Monday through Jan 3, as a reminder and shortly thereafter will transition away from the old database to the new one.
Thanks in advance for your patience and cooperation.
Nestor Your Friendly Neighborhood SysOp.
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 23:31:20 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jmo12505-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, December 20, 2004 at 22:33:37 ---------------------------------------------------------------------------
Email: jmo12505-at-yahoo.com Name: Judy Ogilvie
Organization: Saint Louis University
Title-Subject: [Microscopy] [Filtered] LM - advice on embedding plastic
Question: I have been using Epon Araldite plastic for many years for both EM & LM (1 micron) sectioning. I am gearing up for a new project that will involve embedding and sectioning of a fairly large number of mouse brains. I will not need to do any EM on this tissue. I also have undergraduates who are interested in participating in the project but have no prior histology experience. Are any of the newer plastics easier to work with for embedding and sectioning?
My school has a Jeol JEM 2010 TEM running at 200kV. Thanks a lot!
And the rest:
Very grateful for giving me so much useful advices! I am currently waiting my turn for next session of TEM to try and experiment again!!!!! Will update any result if possible!
Regards, Yee Yan Nanyang Technological University
--- Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} wrote:
} You may want to repost } with the make and model } number of your TEM. } In that manner, people } can give you specific } answers. } } regards, } } Jim } } } } From MicroscopyL-request-at-ns.microscopy.com Sat } Dec 18 02:27:48 2004 } } X-Authentication-Warning: ns.microscopy.com: mail } set sender to MicroscopyL-request-at-ns.microscopy.com } using -f } } Date: Sat, 18 Dec 2004 12:59:01 +0800 (CST) } } From: Tay Yee Yan {one_twinklestar-at-yahoo.com.sg} } } Subject: [Microscopy] Diffraction Pattern for } nano-particle/quantum dots in TEM } } To: Microscopy-at-msa.microscopy.com } } MIME-Version: 1.0 } } Content-Type: text/plain; charset=iso-8859-1 } } Content-Transfer-Encoding: 8bit } } } } } } } } } ------------------------------------------------------------------------------ } } 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: } } } } I am a beginner in using TEM. I am currently in } } investigating } } nano-particles of 20nm and below. However, I have } } difficulty in getting the } } diffraction pattern in TEM. May I know if i can } have } } some advice on this } } issue? Thanks a lot! } } } } Regards, } } Yee Yan } } Postgraduate } } Nanyang Technological University } } Singapore } } } } } __________________________________________________ } } Do You Yahoo!? } } Download the latest ringtones, games, and more! } } http://sg.mobile.yahoo.com } } }
__________________________________________________ Do You Yahoo!? Log on to Messenger with your mobile phone! http://sg.messenger.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 21 05:45:05 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (neil-at-young8696.freeserve.co.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 21, 2004 at 05:52:35 ---------------------------------------------------------------------------
Email: neil-at-young8696.freeserve.co.uk Name: Neil P. Young
Organization: University of Birmingham, England
Title-Subject: [Microscopy] [Filtered] STEM and beam induced contamination
Question: Dear all, I am a new TEM/STEM user. My machine is an FEI Tecnai F20 with STEM facility operating at 200kV. I am interested in looking at metal nanoparticles, diameter {20nm. I have deposited such particles (Au) upon carbon support films (Agar Scientific). I am having many problems with beam induced contamination of the carbon support, making imaging of the nanoparticles impossible with STEM. I am investigating ways of cleaning the supports before depositing nanoparticles. I wonder if such contamination problems are common with STEM and would be interested in possible solutions to the problem. I am currently investigating cleaning with organic solvents and heating or irradiation with UV. regards
There are those much better qualified to answer this than me but I'll give an answer as a 'neighbour' with some experience of it.
Yes, contamination is a problem with STEM. It is with all small probe modes, especially as you use higher and higher beam currents. You will see contamination in a FEG STEM when your specimen looks OK in every other machine.
First you need to identify the source of the contamination. It is, as you say, most likely to be from your specimen. Do other users have contamination free STEM sessions with the same holder? If so then you can rule out the column or holder cleanliness, if not then maybe there is a problem there.
If you 'flood' the specimen does it reduce the contamination rate for a while? If so then the major contamination source is your specimen, if not the contamination is coming from the vacuum. To flood the specimen use a large C aperture and spot size and keep the beam on an area larger than a grid square (~200um dia) for 15 minutes. Check the contamination rate in the centre of the area before and after flooding.
If it is the C filmed grids giving rise to the contamination we have found that annealing them in a (clean) vacuum is a very efficient way of cleaning them, about 200C for 10-15 minutes is usually enough. The Cu tends to get annealed if the temperature is too high, not a real problem it just makes handling them more difficult.
Plasma cleaning can also be used, if you have access to a plasma cleaner, but it will also dissolve your film.
Ensure your method of depositing gold is clean or else you've wasted your time cleaning the C film. If is is a diffusion pumped vacuum then it should be trapped to prevent backstreaming of oil. If it is a chemical or solvent then it needs to be clean and pure, fresh from the container into cleaned glass not from a plastic washbottle.
Ensure your method of storing specimens is clean. Avoid contact with plastic containers - gelatine capsules should be thrown away and never used. Keep specimens under clean vacuum or in a dry container.
If no-one has clean specimens and flooding does not improve the contamination rate it is probably your specimen holder (clean it properly) or column (bake it out) causing the problem.
If the contamination rate from your sample is small you may be able to use the flooding method to prevent contamination build up for long enough to run your experiment.
Good luck, Ron
On Tue, 21 Dec 2004 08:27:37 -0600 by way of MicroscopyListserver {neil-at-young8696.freeserve.co.uk} 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 } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (neil-at-young8696.freeserve.co.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 21, 2004 at 05:52:35 } --------------------------------------------------------------------------- } } Email: neil-at-young8696.freeserve.co.uk } Name: Neil P. Young } } Organization: University of Birmingham, England } } Title-Subject: [Microscopy] [Filtered] STEM and beam induced contamination } } Question: Dear all, I am a new TEM/STEM user. My machine is an FEI Tecnai F20 with STEM facility operating at 200kV. I am interested in looking at metal nanoparticles, diameter {20nm. I have deposited such particles (Au) upon carbon support films (Agar Scientific). I am having many problems with beam induced contamination of the carbon support, making imaging of the nanoparticles impossible with STEM. I am investigating ways of cleaning the supports before depositing nanoparticles. I wonder if such contamination problems are common with STEM and would be interested in possible solutions to the problem. I am currently investigating cleaning with organic solvents and heating or irradiation with UV. } regards } } Neil Young } University of Birmingham } } --------------------------------------------------------------------------- } }
---------------------- 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 Tue Dec 21 15:42:50 2004
Hi all, I'm posting these job announcements for a friend, Robby Roberson. Please contact Robby (not me) if you need more information. robby2-at-asu.edu
happy holidays, Beth ***** As you may know there are three bioimaging positions in the School of Life Sciences at Arizona State University (ASU): 1) Bioimaging Laboratory Director to be appointed as a tenure-track Associate or Full Professor, 2) Bioimaging Lab Manager (2 positions): responsible for user training/interaction, equipment maintenance, etc. (position announcement attached as advertised in Science).
The W.M. Keck Bioimaging Laboratory and the Life Sciences Electron Microscopy Laboratory at Arizona State University will soon be merged and administered by a new faculty Director and two Laboratory Managers. For Director we are intending to appoint a senior, tenured faculty member who maintains an active, well funded research program in cell or molecular biology, preferably a program that incorporates bioimaging techniques as an important feature. The other fifty percent of the Directors workload, including all teaching and service, would be directed toward the bioimaging laboratory. Teaching would likely consist of bioimaging courses and laboratories at the graduate level although there is flexibility here. Service would include management and development of the bioimaging laboratory. These activities would likely include supervision of two laboratory managers, preparation of community-use equipment grants for external funding agencies, organization of bioimaging workshops, development of laboratory policy, and interaction with other units and research programs that use the bioimaging laboratory. We hope to hire a Director that sees development of the Bioimaging Laboratory as being a means to strengthen their own research program.
The Lab Managers (service professionals in ASU terminology) will oversee the daily activities of the laboratory, with the anticipation that one manager will have expertise in electron microscopy and the other expertise in light microscopy. The Bioimaging Laboratories are a University-wide resource and as such the personnel hired must be capable of interacting with faculty, staff, students and collaborating scientists from a broad range of fields.
As stated in the ad, the Laboratory currently provides bioimaging services in electron microscopy, cryofixation, freeze fracture, laser-scanning confocal microscopy (single and multi-photon), video microscopy, live cell imaging, single-molecule fluorescence microscopy, and atomic force microscopy. ASU has an established history of excellence in imaging with its Center for High Resolution Electron Microscopy and its Centers for Optical Biotechnology and Molecular Biophysics. This present hiring initiative is meant to maintain and strengthen this emphasis in imaging. ASU is currently expanding in biomedical and applied molecular research with the formation of the Biodesign Institute and in growing relationships with Phoenix-area biomedical and agricultural research institutes.
Formal applications can be sent to: Chair, Bioimaging Search Committee, School of Life Sciences, PO Box 874501, Arizona State University, Tempe, AZ 85287-4501. Initial closing date for applications is December 28, 2004; if not filled, subsequent applications will be reviewed weekly thereafter until the search is closed.
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"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)
There are some articles and a Book Chapter I wrote at that site. A number of Vendors also market plasma cleaners including: SouthBay Technology, XEI, Fischione, and SPI.
Disclaimer:
ANL/University of Chicago holds the original patent on Plasma Cleaning for AEM applications and I did a modicum of that work.
Nestor Your Friendly Neighborhood SysOp
-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov NEESLab: http://neestpm.mcs.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
===========================================
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 06:20:57 2004
I am glad to see a resolution to a topic that I too thought it was rather harshly or inappropriately attacked.
Dear Dr. John Russ as Scott as well stated I too look to you for answers when it comes to image analysis. I think the country looks to you for image analysis. I have taken image analysis courses from you. Again as Scott said I see your name on the listserver, I read it carefully. In our Microscopy Lab I have two colleagues that have taken microscopy course/s from a legendary man named Dr. Walter McCrone and they thought of him as if the Webster Dictionary described the word microscopy as Dr. Walter McCrone. So, it sounds like I am not alone to think that you must be a description in the Webster Dictionary under image analysis. However, it takes people such as you to bring out the best in the rest of us. I have yet to publish any paper/article related to image analysis just because of being afraid to make a mistake. I am glad to see the professionalism in you to re-assure the rest of us (at least me) that mistakes can be made and maybe even corrected in a professional manner. I assure you that this is to remind myself and others like me that one can write articles/papers on image analysis and not be ridiculed by the community.
I believe that a great teacher goes on teaching even when he/she is not teaching.
Merry Christmas and Happy New Year to all.
Nicola Ranieri, Microscopy/Image Analysis US FDA Forensic Chemistry Center 6751 Steger Drive Cincinnati, Ohio 45237-3097 (513) 679-2700 X253 (513) 679-2761 FAX nranieri-at-ora.fda.gov
-----Original Message----- } From: Walck, Scott D. [mailto:walck-at-ppg.com] Sent: Friday, December 17, 2004 4:04 PM To: MicroscopyListserver (E-mail)
I'm sorry John, but I can't let this slip away.
First, let me say that I do not have any issues with the technical issues that you put in response to Jerry's article because I do not consider myself an expert and do consider you one. When you write an article or submit something on the listserver, my ears perk up and I read it. I have personally benefited from your generosity in discussing and helping me with stereological problems. However, there are a number of points that you raise that I think that should be addressed.
You are correct that MT is not a refereed journal, but I don't think that your assessment that people don't take it seriously is correct. Ron is an experienced microscopist and does a very good job of putting solid articles together in a coherent fashion. For the most part, the articles are by people that are well-respected. I have written a few articles for MT (not that I am well-respected, LOL) and I am careful to try not to put anything that is incorrect in there because I realize that it is not refereed. It also takes a bit of time to put an article together and I thank the efforts of those who have written for MT. I have talked to others who have written articles and for the most part, they have taken similar care. If you have technical exceptions to an article that someone has written, I would bet that Ron would be willing to include those if they were written in an objective mode.
Next, the Listserver is for discussion. Your points with respect to the MT article are valid and are on subject and definitely should generate discussion. However, I think that your delivery was a bit personal and rather scathing. That wasn't appropriate for the listserver and was against the rules. John, forgive me for saying this, but it looks unprofessional and lacked tact. It looked as if you were frustrated with nobody listening to you. Believe me that that is not the case when you talk about imaging. I just was not comfortable reading your submission because of the personal attack that it had.
I wrote this because I don't want people afraid to submit either to the listserver or MT for fear of personal attacks by well-known people in the field. I hope that this has not jeopardized our friendship in any way. These are my opinions alone.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472 Walck-at-PPG.com (412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Friday, December 17, 2004 12:20 PM To: Microscopy-at-msa.microscopy.com
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When Jerry Sedgewick's articles first appeared in Microscopy Today, I wrote a lengthy correction to some of his misinformation and sent it to Ron Anderson. There was no reply, and nothing appeared in the magazine, but Ron later told
me he had forwarded the letter to Jerry, who has never responded. Realizing that the magazine is not a referreed journal and probably isn't taken very seriously anyway, I put the continuing errors in the series of artices in the category of a minor annoyance.
But in the most recent issue, at the end of Jerry's article, he has inserted
the statement 'Note: More thorough explanations of the concepts presented in
this article can be found in "The Image Processing Handbook" by John Russ.' By invoking my name and book in an apparent attempt to gain some credibility for the article, which is almost entirely incorrect and misleading, that statement requires an answer.
First, the cited book does not discuss the subject of matching the appearance of images on printouts and the computer screen. I have written in another book on that topic, but the bible on the subject can be found in David Blatner and Bruce Fraser's "Real World Photoshop" (Peachpit Press). Another excellent choice for anyone concerned about doing this right is to get either the GretagMacbeth Eye-One (my preference) or ColorVision Spyder calibration hardware and software, and follow the straightforward procedures which will make sure that your display, printout, and even projector all show the same thing.
The Sedgewick article is completely wrong (as usual). The International Color Consortium begun by companies like HP and Apple has constructed a technically sound and easily implemented framework that allows manufacturers of software
(e.g., Photoshop), hardware (display cards, monitors, printers) and consumables (ink, paper) to provide widely available curves that everyone should use. Bypassing this (per Jerry's recommendation) and attempting to adjust the image or the monitor to make images more dramatic is a bad idea. Altering the contrast of "scientific" images as compared to normal photography is nonsense. Ink jet printers are fine and have their place, but are certainly not the only way to get good output. The article has plenty of other, more subtle errors (e.g., the dialog shows that sRGB color space is being used, instead of one such as
Adobe 1998 with its greater gamut).
Ignore the article. In fact, ignore all of the Sedgewick articles about Photoshop. Even in those cases where the instructions are not wrong, they are not the best way to accomplish a given result. Go read a good book. There are lots of them, written by people who actually know what they are writing about.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 09:11:48 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (R_Chen-at-fccc.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, December 22, 2004 at 09:00:18 ---------------------------------------------------------------------------
Question: We are trying to image DNA and dsRNA using Cytochrom C or BAC spreding/rotary shadowing techniques. While we were ble to reach reasonable contrast, we still have problems with longer DNA (say, 40 kb) - despite our efforts, the filaments usually remind more a complex bundle of knotted string than anything else. Any bright suggestions?
John Russ writes ... regarding the MT Sedgewick article
} [...[ } the dialog shows that sRGB color space is being used, } instead of one such as Adobe 1998 with its greater gamut. } } Ignore the article. [...] Go read a good book. There are } lots of them, [...]
Just as another note regarding Photoshop and color spaces (e.g., sRGB, AdobeRGB). I would suggest another book, "Real World Color Management" (Fraser, et al, Peach Press). Scientific imagery and Photoshop don't necessary mix because recent versions of PS will insist on a working color space (i.e., a work-around isn't easy to find). Given a general installation of PS, and PS's default insistance on a working space, you can end up with different color, or you can end up with different RGB data. Either of the "Real World" books will make you aware of, and how to avoid either, depending on the task at hand.
The general implication is understanding Photoshop is a significant study. Unless you know what you're doing, you should use PS for presentation only .. while keeping your original image files archived and intact.
Happy Holidays!!! ...
my CA$0.02 & cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 11:05:43 2004
In a message dated 12/22/04 11:46:59 AM, michael-at-shaffer.net writes:
} Just as another note regarding Photoshop and color spaces (e.g., sRGB, } } AdobeRGB). I would suggest another book, "Real World Color Management" } } (Fraser, et al, Peach Press).
I agree with Michael that all of Bruce Fraser's books on Photoshop are excellent sources of correct information. Another good one to consider is Michael Kieran "Photoshop Color Correction" (Peachpit Press). I refer to the Kieran and Fraser books regularly. Somewhat different in focus is Dan Margulis "Professional Photoshop" (Wiley) which is mostly concerned with CMYK color spaces used in printing but offers a useful point of view.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 12:17:08 2004
Plasma Cleaning of your support films would be the answer. I have some documentation on this being done with our PC2000 Plasma Cleaner and would be pleased to send it along to you if you think it would be useful. Please contact me off-line for details.
Best regards-
David
-- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- {http://www.msa.microscopy.com/MicroscopyListserver http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by ( {mailto:neil-at-young8696.freeserve.co.uk neil-at-young8696.freeserve.co.uk) from {http://microscopy.com/MicroscopyListserver/MLFormMail.html http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 21, 2004 at 05:52:35 } --------------------------------------------------------------------------- } } Email: {mailto:neil-at-young8696.freeserve.co.uk neil-at-young8696.freeserve.co.uk } Name: Neil P. Young } } Organization: University of Birmingham, England } } Title-Subject: [Microscopy] [Filtered] STEM and beam induced contamination } } Question: Dear all, I am a new TEM/STEM user. My machine is an FEI Tecnai F20 with STEM facility operating at 200kV. I am interested in looking at metal nanoparticles, diameter {20nm. I have deposited such particles (Au) upon carbon support films (Agar Scientific). I am having many problems with beam induced contamination of the carbon support, making imaging of the nanoparticles impossible with STEM. I am investigating ways of cleaning the supports before depositing nanoparticles. I wonder if such contamination problems are common with STEM and would be interested in possible solutions to the problem. I am currently investigating cleaning with organic solvents and heating or irradiation with UV. } regards } } Neil Young } University of Birmingham } -- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
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From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 12:19:51 2004
A very thorough response and some excellent suggestions. One correction that I would like to make is that carbon support films can be cleaned in a plasma cleaner without dissolving the film. In fact, this was tested on our plasma cleaner and documented by our friendly neighborhood sysop - Nestor Zaluzec. Perhaps the plasma cleaner you used did not have the flexibilty to adjust the parameters appropriately to clean without dissolving the film. I am away from the office now, but when I return I would be happy to send you the reference if you have an interest. In any case, it most certainly can be done with our PC2000.
Best regards-
David
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com Ron Doole 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 Neil, } } There are those much better qualified to answer this than } me but I'll give an answer as a 'neighbour' with some } experience of it. } } Yes, contamination is a problem with STEM. It is with all } small probe modes, especially as you use higher and higher } beam currents. You will see contamination in a FEG STEM } when your specimen looks OK in every other machine. } } First you need to identify the source of the contamination. } It is, as you say, most likely to be from your specimen. Do } other users have contamination free STEM sessions with the } same holder? If so then you can rule out the column or } holder cleanliness, if not then maybe there is a problem } there. } } If you 'flood' the specimen does it reduce the } contamination rate for a while? If so then the major } contamination source is your specimen, if not the } contamination is coming from the vacuum. To flood the } specimen use a large C aperture and spot size and keep the } beam on an area larger than a grid square (~200um dia) for } 15 minutes. Check the contamination rate in the centre of } the area before and after flooding. } } If it is the C filmed grids giving rise to the } contamination we have found that annealing them in a } (clean) vacuum is a very efficient way of cleaning them, } about 200C for 10-15 minutes is usually enough. The Cu } tends to get annealed if the temperature is too high, not a } real problem it just makes handling them more difficult. } } Plasma cleaning can also be used, if you have access to a } plasma cleaner, but it will also dissolve your film. } } Ensure your method of depositing gold is clean or else } you've wasted your time cleaning the C film. If is is a } diffusion pumped vacuum then it should be trapped to } prevent backstreaming of oil. If it is a chemical or } solvent then it needs to be clean and pure, fresh from the } container into cleaned glass not from a plastic washbottle. } } Ensure your method of storing specimens is clean. Avoid } contact with plastic containers - gelatine capsules should } be thrown away and never used. Keep specimens under clean } vacuum or in a dry container. } } If no-one has clean specimens and flooding does not improve } the contamination rate it is probably your specimen holder } (clean it properly) or column (bake it out) causing the } problem. } } If the contamination rate from your sample is small you may } be able to use the flooding method to prevent contamination } build up for long enough to run your experiment. } } Good luck, } Ron } } On Tue, 21 Dec 2004 08:27:37 -0600 by way of } MicroscopyListserver {neil-at-young8696.freeserve.co.uk 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 } } ------------------------------------------------------------------------------- } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (neil-at-young8696.freeserve.co.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 21, 2004 at 05:52:35 } } --------------------------------------------------------------------------- } } } } Email: neil-at-young8696.freeserve.co.uk } } Name: Neil P. Young } } } } Organization: University of Birmingham, England } } } } Title-Subject: [Microscopy] [Filtered] STEM and beam induced contamination } } } } Question: Dear all, I am a new TEM/STEM user. My machine is an FEI Tecnai F20 with STEM facility operating at 200kV. I am interested in looking at metal nanoparticles, diameter {20nm. I have deposited such particles (Au) upon carbon support films (Agar Scientific). I am having many problems with beam induced contamination of the carbon support, making imaging of the nanoparticles impossible with STEM. I am investigating ways of cleaning the supports before depositing nanoparticles. I wonder if such contamination problems are common with STEM and would be interested in possible solutions to the problem. I am currently investigating cleaning with organic solvents and heating or irradiation with UV. } } regards } } } } Neil Young } } University of Birmingham } } } } --------------------------------------------------------------------------- } } } } } } } } } } } } } } ---------------------- } 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/ } ********************************* } } } } } } } } }
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
---------------------- 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/ *********************************
-- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 13:21:04 2004
I would like to add my two cents into this discussion regarding carbon contamination. This is in regard to observations that I have had using carbon films, FEG TEMs, and plasma cleaners. I have anecdotal information on carbon films and plasma cleaning of coated samples that I would like to share.
General Claim: The type of carbon that you have is going to play a role in "contamination" effects and the effectiveness in carbon removal with plasma cleaners.
When I was working at Wright Patterson Air Force Base, we were looking at pulsed laser deposited (PLD) diamond-like carbon (DLC) films. First, it is well-known in the literature that the way the DLC films are prepared and their inherent chemistry plays a major role in their tribological properties as well as their reaction to environmental factors. The films that we grew by PLD showed a layered pattern of alternating regions of amorphous DLC that varied in the ratio of sp3 to sp2 bonds, i.e. the darker bands were more diamond-like and the lighter bands were more graphite-like. When a focused FEG probe was used to acquire EELS spectra, the dark bands would show all SP3 bonding initially but immediately start showing or growing the SP2 structure during beam exposure within seconds. The light bands would do it also, but the rate of build-up was comparatively slower (never measured -only observed). Since the films were grown on silicon substrates, I prepared these samples by the small angle cleavage technique. This gave great, very thin carbon films, and SACT is very good for minimizing, if not eliminating, contamination problems. If the beam was not converged on the PLD-DLC coating, but was used in the normal imaging mode, no contamination build-up was seen, regardless of how much time the sample was under the beam. If the converged beam was moved to the substrate, the beam could be in position for up to 5 minutes with no noticeable contamination. I concluded that the beam was converting the carbon type when converged on the coating and that it was not contamination.
When I first came to PPG, we had a JEOL 1200EX that operated at 120 kV. At this voltage, the glass is softened because of the energy that is deposited in the glass. When the samples were taken to a 200 or 300 kV machine (outside lab with FEG or LaB6), we did not see the softening effect. To help the problem when imaging at 120 kV, I found that a very light evaporated carbon layer helped dissipate the heat as well as eliminate charging effects. The evaporator was a diffusion pumped system and pretty dirty. I worried about contamination problems in the FEG machines if I used the samples coated with this carbon in these better FEG machines. To help minimize charging in samples taken to other sites with FEG, I developed a way of sputter coating carbon films destined for use in the good machines with my Bal-Tec RES 100 ion mill. Well the situation arose where I didn't have one of the "clean" carbon sputtered samples and only had the 120 kV carbon evaporated films. I thought that I could plasma clean (EAF Fishcione unit) the samples for a very short time and partially remove the carbon and have a light, but sufficiently conducting film for the FEG use. What I found was that the evaporated carbon resisted removal by plasma cleaning. Playing around a bit, I found that the sputter coated carbon films could be removed by plasma cleaning.
There are other behavior of carbon films with plasma cleaners that was written about in several publications in which I was a co-author. Those publication are available at the South Bay Technology web site for downloading as was mentioned by another posting. I think that an image of the SACT prepared PLD-DLC film on silicon with the EELS spectra is also available from their site.
My point in submitting these observations is that the behavior of the carbon films both under the beam and with plasma cleaning will differ depending on the state of the carbon which is determined by the deposition parameters.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472 Walck-at-PPG.com (412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Ron Doole [mailto:ron.doole-at-materials.oxford.ac.uk] Sent: Tuesday, December 21, 2004 11:58 AM To: by way of MicroscopyListserver Cc: microscopy-at-microscopy.com
Dear Neil,
There are those much better qualified to answer this than me but I'll give an answer as a 'neighbour' with some experience of it.
Yes, contamination is a problem with STEM. It is with all small probe modes, especially as you use higher and higher beam currents. You will see contamination in a FEG STEM when your specimen looks OK in every other machine.
First you need to identify the source of the contamination. It is, as you say, most likely to be from your specimen. Do other users have contamination free STEM sessions with the same holder? If so then you can rule out the column or holder cleanliness, if not then maybe there is a problem there.
If you 'flood' the specimen does it reduce the contamination rate for a while? If so then the major contamination source is your specimen, if not the contamination is coming from the vacuum. To flood the specimen use a large C aperture and spot size and keep the beam on an area larger than a grid square (~200um dia) for 15 minutes. Check the contamination rate in the centre of the area before and after flooding.
If it is the C filmed grids giving rise to the contamination we have found that annealing them in a (clean) vacuum is a very efficient way of cleaning them, about 200C for 10-15 minutes is usually enough. The Cu tends to get annealed if the temperature is too high, not a real problem it just makes handling them more difficult.
Plasma cleaning can also be used, if you have access to a plasma cleaner, but it will also dissolve your film.
Ensure your method of depositing gold is clean or else you've wasted your time cleaning the C film. If is is a diffusion pumped vacuum then it should be trapped to prevent backstreaming of oil. If it is a chemical or solvent then it needs to be clean and pure, fresh from the container into cleaned glass not from a plastic washbottle.
Ensure your method of storing specimens is clean. Avoid contact with plastic containers - gelatine capsules should be thrown away and never used. Keep specimens under clean vacuum or in a dry container.
If no-one has clean specimens and flooding does not improve the contamination rate it is probably your specimen holder (clean it properly) or column (bake it out) causing the problem.
If the contamination rate from your sample is small you may be able to use the flooding method to prevent contamination build up for long enough to run your experiment.
Good luck, Ron
On Tue, 21 Dec 2004 08:27:37 -0600 by way of MicroscopyListserver {neil-at-young8696.freeserve.co.uk} 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 } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (neil-at-young8696.freeserve.co.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 21, 2004 at 05:52:35 } --------------------------------------------------------------------------- } } Email: neil-at-young8696.freeserve.co.uk } Name: Neil P. Young } } Organization: University of Birmingham, England } } Title-Subject: [Microscopy] [Filtered] STEM and beam induced contamination } } Question: Dear all, I am a new TEM/STEM user. My machine is an FEI Tecnai F20 with STEM facility operating at 200kV. I am interested in looking at metal nanoparticles, diameter {20nm. I have deposited such particles (Au) upon carbon support films (Agar Scientific). I am having many problems with beam induced contamination of the carbon support, making imaging of the nanoparticles impossible with STEM. I am investigating ways of cleaning the supports before depositing nanoparticles. I wonder if such contamination problems are common with STEM and would be interested in possible solutions to the problem. I am currently investigating cleaning with organic solvents and heating or irradiation with UV. } regards } } Neil Young } University of Birmingham } } --------------------------------------------------------------------------- } }
---------------------- 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 Dec 23 05:41:10 2004
Dear listers, I now have a working Micrion 2500 FIB system (lovely) and made my first TEM specimen yesterday, a nice Christmas present! I am now looking at micromanipulators for lift-out TEM specimen prep. I would be very grateful to hear from anyone who has recently purchased a micromanipulator for this purpose, either in-situ or ex-situ, what are the pros and cons of each, and of course how much it is likely to cost. Manufacturers are welcome to respond off-line.
Although not microscopy-related per se, I have a Philips HRD five-crystal X-ray diffraction kit for disposal. The HT generator is no longer working (I can give more details to anyone who is interested). A very nice machine with a good solid goniometer. It would be nice to find a home for this, as usual I hate to see good machines going into landfill. I don't want any money for it, but if you take it you will pay for any shipping costs.
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited. No part of this message can be considered a request for goods or services. =======================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 23 08:27:17 2004
8th InterAmerican Congress of Electron Microscopy,
will be held in Havana, Cuba, from 25 to 30 September 2005.
Despite the restrictions on travel to Cuba by citizens and residents of the USA, full-time professional microscopists are allowed to travel to Cuba for this meeting. Details are given along with other valuable information on the conference web site:
http://ciasem2005.cigb.edu.cu/
Bookmark this site, which will be kept up to date as new information becomes available.
The conference is sponsored by CIASEM, the InterAmerican Committee of Electron Microscopy Societies http://www.ciasem.com/
.......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 23 11:40:27 2004
richard, we have a fei db235, in service for about 3 years. we purchased the in-situ omniprobe plucker with the initial installation. it works well, but is somewhat tricky to align the tips. in our version, the coordinate system for the plucker and the sample are different, meaning a z move with the plucker, moves x,y,z, in relation to the sample. i think omniprobe has a much better system available, but at a cost. ours was around $80K as i recall, much more for their newer system. we have recently acquired a zyvex s100 manipulator for in-situ probe measurements, manipulation, and tem lift out. priced at $180K. it was just installed, and we have not fully tried it, but it looks very nice. with either system we would like to have sharper probe tips. we currently use two sources for tips with the omniprobe: micromanipulator company, 7x (.1 micron radius) and omniprobe tips.
mike
Michael T. Marshall Research Engineer University of Illinois Frederick Seitz Materials Research Laboratory 104 South Goodwin Avenue Urbana, IL 61801 Voice: 217/265-5380 Fax: 217/244-2278 marshall-at-mrl.uiuc.edu
-----Original Message----- } From: Richard Beanland [mailto:richard.beanland-at-bookham.com] Sent: Thursday, December 23, 2004 5:40 AM To: Microscopy Listserver (E-mail)
Dear listers, I now have a working Micrion 2500 FIB system (lovely) and made my first TEM specimen yesterday, a nice Christmas present! I am now looking at micromanipulators for lift-out TEM specimen prep. I would be very grateful to hear from anyone who has recently purchased a micromanipulator for this purpose, either in-situ or ex-situ, what are the pros and cons of each, and of course how much it is likely to cost. Manufacturers are welcome to respond off-line.
Although not microscopy-related per se, I have a Philips HRD five-crystal X-ray diffraction kit for disposal. The HT generator is no longer working (I can give more details to anyone who is interested). A very nice machine with a good solid goniometer. It would be nice to find a home for this, as usual I hate to see good machines going into landfill. I don't want any money for it, but if you take it you will pay for any shipping costs.
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited. No part of this message can be considered a request for goods or services. =======================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 23 18:18:59 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jfb-at-yadtel.net) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, December 23, 2004 at 15:29:38 ---------------------------------------------------------------------------
Question: I have a Zeiss Tessovar lens turret that has what appears to be either optical adhesive or some previous cleaning solution that is slightly clouding the back surface (very edges) of a small .25 and .5 lens. Is there a method of releasing the glue on the front lens(of two sandwich with space between)in order that one could access the middle surfaces to clean, and then replace? Any comments would be appreicated.
Scott Walck wrote: ============================================================================ = } snip {
General Claim: The type of carbon that you have is going to play a role in "contamination" effects and the effectiveness in carbon removal with plasma cleaners.
} snip { ============================================================================ = Since SPI Supplies also manufactures a plasma cleaner under license from Argonne National Laboratory, see URL http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml
I would like to make the following comments:
a) The efficacy of the cleaning depends on the power being used for the cleaning, with 100 watts being much more effective than 10 watts. The higher power units would also be much more aggressive in terms of etching away a carbon filmed grid whereas 10 watts, so far as I can determine, would not etch away a carbon filmed grid. Units described as plasma cleaners generally operate at 10 watts or less, plasma etchers at 100 watts or more. One can clean with an etcher in some circumstances, with oxygen, where there are no carbon inclusions or other carbonaceous domains in the sample. One would not want to clean with Ar at 100 watts since those conditions would argon etch the rod and holder if not also the sample itself.
b) One could use a SiO2 filmed grid in many of the instances one would be using a carbon filmed grid. One of the main reasons why one would prefer to use an SiO2 filmed grid is that the grid can be "cleaned" with pure oxygen without any fear of the substrate being etched away. The SiO2 filmed grids will generally withstand a more aggressive cleaning at higher power levels. Exposure to the higher power level generally results in a longer contamination-free observation time.
c) We have never seen instances where some carbonaceous contamination formed in a TEM on a TEM foiled sample can be removed this way and other contamination could not (because it was a DLC).
Disclaimer: SPI Supplies manufactures the Plasma Prep Plasma Cleaner instrument and we are also producers of both carbon and SiO2 filmed grids.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 25 10:45:17 2004
During the Xmas holidays as some of you know , I am deploying a new database and restructuring the system. In order to make sure everyone stays on-line, there will be some overlap of the databases. This means that a limited number of you may receive duplicate copies of Email postings if your address is listed twice (once in each database). This should occur only if your address is subscribed twice in two different forms something like this.
This usually occurs when individuals setup aliases and forwarding accounts.
If both databases have the same (i.e. identical) Email addresses then duplicate should NOT occur, this case should apply to the MAJORITY of subscribers.
Purging these pseudo-duplicates (remember the software doesn't apriori know these are the same individual) is time consuming and tedious. I'm asking that if you are one of these individuals who gets duplicates please bear with them until the ~ first week of Jan, when they should stop. I expect the volume of such duplicated Email over the Xmas holiday to be pretty minimal as traffic dies down alot over the holiday.
If you find this problem annoying, just unsubscribe the address which you no longer use by the online form (http://www.microscopy.com/MicroscopyListserver).
Remember you can only post messages from an address which is recorded in the subscription list, so if you make a change, insure that your EMail client is also using the correct "FROM" address in its settings file.
Thanks in advance... & have a good holiday
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 25 16:04:52 2004
} {...snip...} } } c) We have never seen instances where some carbonaceous contamination } formed in a TEM on a TEM foiled sample can be removed this way and other } contamination could not (because it was a DLC). }
Hi Chuck,
I'm unclear about what you are saying in point (c). Are you saying that e-beam generated contamination is DLC or that the other material is DLC? Can DLC be removed by plasma cleaning? Do you have any references on the form of carbon created by the e-beam? Presumably, SEM raster squares are the same material as TEM contamination, yet they do eventually clean up with plasma cleaning.
SiO2 filmed grids sound like a good solution when you need to plasma clean. Are there any issues with the support film conductivity? I'd hate to have the film blow apart like uncoated formvar! Also, has anyone used the SiO2 support films for holding FIB liftout samples?
Cheers, Henk
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 25 22:18:59 2004
Hendrik O. Colijn wrote: ========================================================= Hi Chuck,
I'm unclear about what you are saying in point (c). Are you saying that e- beam generated contamination is DLC or that the other material is DLC? Can DLC be removed by plasma cleaning? Do you have any references on the form of carbon created by the e-beam? Presumably, SEM raster squares are the same material as TEM contamination, yet they do eventually clean up with plasma cleaning.
SiO2 filmed grids sound like a good solution when you need to plasma clean. Are there any issues with the support film conductivity? I'd hate to have the film blow apart like uncoated formvar! Also, has anyone used the SiO2 support films for holding FIB liftout samples? ========================================================== Sorry about the ambiguity; I guess that is my punishment for submitting a posting over the holidays.
I was only trying to say that when it comes to contamination formed in a column as a result of interaction with the electron beam, be it an SEM or TEM, it has always been able to be removed with plasma cleaning. I have never encountered what one might call a DLC formed under those conditions. Physics is physics and irrespective of the brand and model of the plasma cleaner being used, or the price paid, the contamination can be removed. The higher the power the faster will be removed the contamination spot but again, that is true for anyone's plasma cleaner.
With regard to SiO2 filmed grids, I have never heard of any conductivity issues, but for FIB liftout samples (ex-situ techniques), take a look at URL http://www.2spi.com/catalog/instruments/perforated-membrane-window-grids. shtml This type of grid might be a more stable alternative to the more traditional holey support films be they carbon or SiO2. And of course, silicon nitride should stand up quite nicely under conditions of plasma cleaning, even a more aggressive cleaning with higher power if the FIB sample could stand it.
Disclaimer: SPI Supplies manufactures the Plasma Prep Plasma Cleaner so we have a vested interest in seeing more people thinking about the plasma cleaning of their samples prior to observation. See URL http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 27 12:18:34 2004
On Dec 25, 2004, at 8:18 PM, Garber, Charles A. wrote:
} With regard to SiO2 filmed grids, I have never heard of any } conductivity } issues,
Dear Chuck, Has anyone investigated SiO2 or SiO conductivities at either LN2 or LHe temps? 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 Dec 27 20:09:46 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rporter1-at-sbcglobal.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, December 27, 2004 at 19:40:24 ---------------------------------------------------------------------------
Question: Hi, Can you recommend manufacturers of reasonably priced microscopes for high school biology? I understand that Motic manufactures good, inexpensive scopes. I would rather not pay the overhead of the big four brands such as Olympus. Can you confirm that we should always purchase achromatic lenses and objectives?
======================================= Second Notice of Microscopy Listserver DataBase Update: ======================================= Dec. 28, 2004
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======================================= First Notice of Microscopy Listserver DataBase Update: ======================================= Dec. 21, 2004
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From MicroscopyL-request-at-ns.microscopy.com Tue Dec 28 12:11:33 2004
On Dec 27, 2004, at 6:12 PM, by way of Ask-A-Microscopist wrote:
} Email: rporter1-at-sbcglobal.net } Name: Randall Porter } } Organization: UCSC } } Education: 9-12th Grade High School } } Location: Santa Cruz, CA, USA } } Question: Hi, } Can you recommend manufacturers of reasonably priced microscopes for } high school biology? I understand that Motic manufactures good, } inexpensive scopes. I would rather not pay the overhead of the big } four brands such as Olympus. Can you confirm that we should always } purchase achromatic lenses and objectives? } } Thanks for your help. } Dear Randall, Caroline Schooley is an expert on this; her contact info and the web site for Project MICRO are:
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
Project MICRO's purpose is to enhance pre-college education, as stated on the web page. 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 Dec 28 20:53:58 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tttan-at-simtech.a-star.edu.sg) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 28, 2004 at 18:47:07 ---------------------------------------------------------------------------
Email: tttan-at-simtech.a-star.edu.sg Name: T T Tan
Organization: Singapore Institute of Manufacturing Technology
Title-Subject: [Microscopy] [Filtered] SEM - Wobbler Adjustment:
Question: Hello,
I am trying to understand the principle of and also to learn about focus wobbler adjustment for SEM. Can anyone please help me? I found the earlier thread in the MSA achieve but couldnít find the article by Le Poole and related articles. Ran searches on Google, Sciencedirect etc. but yielded nothing.
I understand that focus wobbler IN SEM is meant for aperture alignment. How can I tell which alignment (the ìxî or the ìyî) to adjust?
Also earlier on, I was operating the JEOL JSM 6340F. The image couldnít be focused even at 15,000X. When I over focuses, the image smudge in the x-y direction, along bottom left to top right of the screen. How can I adjust this?
} I am trying to understand the principle of and also to learn } about focus wobbler adjustment for SEM. Can anyone please help me? } I found the earlier thread in the MSA achieve but couldnít find } the article by Le Poole and related articles. } Ran searches on Google, Sciencedirect etc. but yielded nothing.
If (1) the final aperture is misaligned, or if (2) the beam is astimagtic, it becomes evident as you focus above and below exact focus ... and this is what the "wobbler" is supposed to help you with ... that is, to continuously focus above/below, while you make the proper adjustments. Some SEM operators, maybe most, have learned not to use it. Depending on the sample, or the degree of the problem, it may be easiest to simply make the adjustments until best focus is achieved.
While the wobbler is enabled, if the image shifts during focus (left-right, up-down, diagonally, ... NOTE twisting is normal), then the problem is the final aperture. Beginning with either aperture adjustment (x,y), I usually adjust the wrong way first to make the problem worse, and then I adjust thru and past the problem, to make it oppositely worse, and then I come back ... and back again ... until I minimize the problem. Do the same with the other aperture adjuster.
If the image does not move, but out-of-focus has directionality (as you describe below), the the problem is astigmatism. The "stigmator" is a bit more difficult to descibe its use. Additionally, there are 2 different types of stigmators used by different manufacturers. I believe the JEOL 6xxx uses "x-y" adjusters and you would approch correcting the problem similar to above. However, if the stigmator is the "rotation-amplitude" type, I find the best approach is to make the problem worse with 'amplitude' .. then minimize the problem with 'rotation' and then minimize the problem with 'rotation'. Reitteration may be required, and keep in mind that your problem may be a combination of (1) and (2).
HTH & Seasons' Greetings from "the rock" cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
} I understand that focus wobbler IN SEM is meant for aperture } alignment. How can I tell which alignment (the ìxî or the ìyî) to adjust? } } Also earlier on, I was operating the JEOL JSM 6340F. The image } couldnít be focused even at 15,000X. When I over focuses, the } image smudge in the x-y direction, along bottom left to top right } of the screen. How can I adjust this? } } } } Regards } T T Tan } } ------------------------------------------------------------------ } --------- } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 29 09:50:05 2004
It's true that correcting astigmatism is difficult to describe without actually having someone observe it. The approach I take, with pretty decent success, is as follows:
Find an area with lots of non-directional detail (i.e., straight edges are bad while "sandy" looking areas are good). Throw the image out of focus until a "smearing" effect is noticed, then go through the focus point and observe that the "smearing" is now shifted 90 degrees from its first orientation. This is definitely asigmatism when this is observed. Still using the focus knob, adjust the image until you are in between the "smearing" parts of the focus range. In other words, the image should now be free of directional distortion, but will probably be somewhat out of focus and if you turn the focus knob significantly in either direction, the smearing will return.
Now use one of the stigmator controls, either x or y, and turn slowly, using it as a "fine focus control". The image should appear to get blurrier or sharper---make it as sharp as you can. Then do the same with the other stigmator control. Repeat the process a couple times, then go back to the focus knob and adjust it back and forth through the focus point. If astigmatism has been corrected, the image will get blurry on both sides of focus, but should not show any directional "smearing".
"Smearing" = the effect you get by putting cooking oil on your hand and wiping it in one direction on your kitchen window. The view of your yard is now blurred and distorted in the direction of your application of the oil. (Make note to clean window before your significant other sees it---explaining this as an education tool is non-effective.)
Happy New Smear!
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.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 29 16:17:29 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cdh1-at-cdc.gov) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, December 29, 2004 at 12:07:22 ---------------------------------------------------------------------------
I was wondering if anyone can assist me in obtaining replacement parts for an LKB 2188 Ultrotome NOVA.
Specific parts include the following. 90 01 3235 Oscillator PC board 90 01 5294 Transformer for diffuse light T1,T2 90 01 3195 Bushing for microscope stand 95 82 1005 Halogen lamp
Also, does anyone know the voltage and wattage for the above designated halogen lamp?
Thanks in advance Charles Humphrey, CDC I may be reached by telephone at 404-639-3307 January 4th 2005 and later
Thanks, Randy and shAf, for your contributions re stigmator and aperture adjustment.
I've never been able to make much sense of the JEOL (840) instructions for condenser lens alignment. It seems they take me into a loop from which there is no escape other than giving up (yet again) in frustration.
Do you guys, or does anyone, have either a lucid step-by-step procedure for this, or an explanation of what is aiming to be achieved that I can translate into specifics for the 840?
I use the thing only as a microprobe, so the spot tightness isn't a real issue, but it would be nice to be able to do the sort of imaging that I know the 840 is capable of, and I suspect that beam current stability is improved by good alignment.
Happy New Year
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 Thu Dec 30 01:30:43 2004
Dear friends, this e-mail is to inform you that you can find
Microscopy Research and Technique, Volume 63 Issue 1 - 1 January 2004 (1 - 86) Special Issue: Two-Photon Microscopy - Part II
as free sample copy on the http://www3.interscience.wiley.com/cgi-bin/jtoc/38527/ webpage.
Please, let me also wish again a bright and peaceful 2005. Alby
------------------------------------------------------------------------ --------------------------- Alberto Diaspro, Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309 URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 30 10:47:41 2004
We have tried SiO2 films in our lab for FIB liftout for just the reason that Chuck mentioned, i.e., that we should be able to then plasma clean without losing the sample. We have tried both bulk and lacy SiO2 films. What we found was that the membrane is sufficiently non-conducting that the liftout membranes charged up, then flew off the grid. Very distressing!!
We tried to cure this by using lacy carbon grids and Cr coating them, but the stress of sputtered Cr broke the membrane. Carbon coating is not an option since plasma cleaning will remove the C.
We have stayed with Formvar membranes and adjusted our plasma cleaning to allow us to minimize contamination without destroying the formvar support film. This was originally driven by the need to plasma clean polymer based low-k dielectrics for chip fabrication; as a side benefit, we found that we preserved the Formvar support film. Using a reduced O2 content in Ar, and reduced power settings, we can do multiple iterations of plasma cleaning. Typically, we find that 20 seconds is sufficient to allow EELS analysis without causing measurable C contamination. I have been able to do three such cleanings on a typical grid without losing the specimen.
Phil
Philip L. Flaitz IBM Microelectronics, Hopewell Junction, NY Ph.......(845) 892-3094, FAX -6256 pager - 800-352-4732, PIN# 1121 flaitz-at-us.ibm.com
"Garber, Charles A." To: MICROSCOPY BB {Microscopy-at-MSA.Microscopy.com} {cgarber-at-2spi.com cc: } Subject: [Microscopy] Plasma cleaning of TEM + SEM samples
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Hendrik O. Colijn wrote: ========================================================= Hi Chuck,
I'm unclear about what you are saying in point (c). Are you saying that e- beam generated contamination is DLC or that the other material is DLC? Can DLC be removed by plasma cleaning? Do you have any references on the form of carbon created by the e-beam? Presumably, SEM raster squares are the same material as TEM contamination, yet they do eventually clean up with plasma cleaning.
SiO2 filmed grids sound like a good solution when you need to plasma clean. Are there any issues with the support film conductivity? I'd hate to have the film blow apart like uncoated formvar! Also, has anyone used the SiO2 support films for holding FIB liftout samples? ========================================================== Sorry about the ambiguity; I guess that is my punishment for submitting a posting over the holidays.
I was only trying to say that when it comes to contamination formed in a column as a result of interaction with the electron beam, be it an SEM or TEM, it has always been able to be removed with plasma cleaning. I have never encountered what one might call a DLC formed under those conditions.
Physics is physics and irrespective of the brand and model of the plasma cleaner being used, or the price paid, the contamination can be removed. The higher the power the faster will be removed the contamination spot but again, that is true for anyone's plasma cleaner.
With regard to SiO2 filmed grids, I have never heard of any conductivity issues, but for FIB liftout samples (ex-situ techniques), take a look at URL http://www.2spi.com/catalog/instruments/perforated-membrane-window-grids. shtml This type of grid might be a more stable alternative to the more traditional holey support films be they carbon or SiO2. And of course, silicon nitride should stand up quite nicely under conditions of plasma cleaning, even a more aggressive cleaning with higher power if the FIB sample could stand it.
Disclaimer: SPI Supplies manufactures the Plasma Prep Plasma Cleaner so we have a vested interest in seeing more people thinking about the plasma cleaning of their samples prior to observation. See URL http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 30 13:17:26 2004
Philip Flaitz wrote: ============================================================= We have tried SiO2 films in our lab for FIB liftout for just the reason that Chuck mentioned, i.e., that we should be able to then plasma clean without losing the sample. We have tried both bulk and lacy SiO2 films. What we found was that the membrane is sufficiently non-conducting that the liftout membranes charged up, then flew off the grid. Very distressing!!
We tried to cure this by using lacy carbon grids and Cr coating them, but the stress of sputtered Cr broke the membrane. Carbon coating is not an option since plasma cleaning will remove the C.
We have stayed with Formvar membranes and adjusted our plasma cleaning to allow us to minimize contamination without destroying the formvar support film. This was originally driven by the need to plasma clean polymer based low-k dielectrics for chip fabrication; as a side benefit, we found that we preserved the Formvar support film. Using a reduced O2 content in Ar, and reduced power settings, we can do multiple iterations of plasma cleaning. Typically, we find that 20 seconds is sufficient to allow EELS analysis without causing measurable C contamination. I have been able to do three such cleanings on a typical grid without losing the specimen. ================================================================ Could you tell us more about the conditions of the plasma cleaning in which you reported that "Carbon coating is not an option since plasma cleaning will remove the C." We have been able to plasma clean carbon support films at 10watts power using oxygen without breaking the carbon films. This is accomplished using the SPI Plasma Cleaner as shown on URL http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml
The application of chromium is still a sputtering process but one can apply a 1 nm (sometimes less) coating of osmium metal in the OPC Osmium Coaters, see URL http://www.2spi.com/catalog/osmi-coat.html I know that the use of the high Z element goes against conventional wisdom but 1 nm is very very thin, so thin in fact that as one can see in URL http://www.2spi.com/catalog/osmium-plasma-coater-demonstration.html its presence at this thin of a layer does not interfere with BSE imaging at all. So if the concept of applying a conductive layer is valid, and chromium did not work, there is a good chance that osmium would work, especially since the heat output with PVD is much less than if by sputtering (or so I have been led to believe). If EELS imaging is being contemplated, and since one is looking through the holes anyhow, the presence of the osmium (or chromium) metal should not matter. We have not actually done these experiments so in that sense this is speculation on my part.
With regard to FIB liftout samples, the perforated Si3N4 membranes might be a solution. I have not heard that charging is a problem but if it was, then a 1 nm layer of osmium on the membrane might solve that problem as well. Or since a perforated Si3N4 membrane film is so much more robust than a lacy carbon film, even chromium might not be a problem if indeed there was a charging problem that had to be solved (this way).
Chuck
Disclaimer: SPI Supplies offers plasma cleaners, osmium coaters and perforated silicon nitride membrane window grids so we would have a vested interest in seeing more people using these products.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ===================================================
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