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Gary Gaugler 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 } ------------------------------------------------------------------------------- } } Would someone point me to the listserver } for failure analysis discussion of metallurgical and } integrated circuit devices? } } tnx, } gary g.
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 2 10:09:20 2004
I've seen hot/cold stages from Deben that look like they could hold a standard pin stub specimen. However, they seem to be optimized for low temperature work. Does anyone know of some other supplier that makes SEM specimen holders that will heat up to about 200C and perhaps cool to -25C?
The unit would need to mate to the stage on a LEO Supra 55VP's specimen interchange stage or on a FEI Sirion 400, under similar circumstances.
Gatan makes a series of heated stages but they seem more for stress testing and bulk specimens. I will have an IC chip thermal expoxy'd to a 12mm diameter Al pin stub. I need to be able to heat this specimen in the SEM and at any tilt and WD that the SEM will support. Temperature stability could be as bad as +-5C. That is OK.
Specimen holder and specimen changeout via slide out chamber door is OK. Specimen interchange lock does not have to be used.
thanks for any ideas and leads, gary g.
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 2 11:27:25 2004
In the latest issue of Microscopy Today there is an article on pseudo-coloring of images. Since this is not something that I normally do, I read the article with interest. I was surprised to find that the author says that biologists nearly always use the "spectrum" color table for the pseudo-coloring of grayscale images. I had thought that those who study visual perception had found that among pseudo-color scales the "thermal" scale is much better than all the others at providing a good intuitive reading of the image. This is, I think, the scale that Photoshop calls "Black Body". Can someone please clear up my confusion. -- .......... 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 Fri Jan 2 11:58:48 2004
Here is the table of contents for the January/February 2004 issue of Microscopy Today.
New Subscriptions via http://www.microscopy-today.com only, please. New subscriptions will close on Thursday 8 January for this issue.
There has been a major change in subscription policies coming out of the MSA Winter Council meeting.
Briefly: Canadians and Mexicans are now offered free subscriptions along with microscopists in the USA.
MSA members anywhere have free subscriptions.
Non-MSA, non-North American subscriptions have been reduced from $80 or $110US to $35US. Additional details in the magazine or on our web site.
THIS WILL BE THE LAST ISSUE NON-MSA MEMBER, NON-NORTH AMERICAN, UNSUBSCRIBED OR NOT-CURRENTLY-SUBSCRIBED INDIVIDUALS WILL RECEIVE!
Listers,
Here is the table of contents for the November/December 2003 issue of Microscopy Today. This issue is mailed with the Call for Papers for the 2004 Microscopy and Microanalysis meeting
New Subscriptions via http://www.microscopy-today.com only, please New subscriptions will close on Tuesday 11 November for this issue.
There has been a major change in subscription policies coming out of the MSA Winter Council meeting.
Briefly: Canadians and Mexicans are now offered free subscriptions along with microscopists in the USA.
MSA members anywhere have free subscriptions.
Non-MSA, non-North American subscriptions have been reduced from $80 or $110US to $35US. Additional details in the magazine or on our web site.
PURGING OF NON-MSA MEMBER, NON-NORTH AMERICAN, UNSUBSCRIBED OR NOT-CURRENTLY-SUBSCRIBED INDIVIDUALS IS UNDERWAY!
January/February 2004 Carmichael: Correlating Fluorescence Microscopy with Electron Microscopy P.E. Batson: Electron Microscopy Enters a New Era Using Aberration __Correction Paula Allan-Wojtas: Microscopy and Imaging of Foods— The Whys and Hows Jerry Sedgewick: Image Stitching Using Photoshop Michael Bode: A Few Thoughts About Image File Storage Paul Beauregard: Behavior of Particle Size Distributions, Means and BET __Values in Ideal and Non-Ideal Morphology Systems in a TEM Katerina Moloni: The Moving Finger Writes: Carbon Nanotubes as AFM Probe __Tips Robert M. Zucker: Confocal Microscopy System Performance: Axial Resolution Shane Roberts, Daniel Flatoff: Enhanced Sample Preparation of Cu Low-k __Semiconductors via Mechanical Polishing and Ion Beam Etching Luc Harmsen: The Year That Was! Microscopy in Southern Africa Robert P. Apkarian: Comments on Cryo High Resolution Scanning Electron __Microscopy Jose A. Mascorro: Propylene Oxide: To Use or Not to Use in Biological Tissue __Processing M. T. Postek & A. E. Vladár: Is Low Accelerating Voltage Always the Best for __Semiconductor Inspection and Metrology?
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 2 12:10:04 2004
Yes it is unfortunately true that many (most?) biologists do use the "spectrum" color scale, largely because it makes "prettier-looking" images. It the cases where they are trying to illustrate quantitative contrast this is not only grossly misleading but it is usually plain wrong and can produce horrific artifacts! The worst offenders are chiefly light microscopists who are trying to represent weak flourescence contrast and for some reason think it shows up "better" with a spectrum scale. In the STEM/X-Ray/EELS biological microanalysis field most of us use some variation of the "black Body" scale which of course more closely parallels the greyscale that is intuitively quantitative anyway (black = 0, shades of grey through white represent more positive values). Relative contrast or non-linear scaling can be achieved by manipuating the scale either continuously or by introducing discontinuities to other scales; of course color then becomes essential (a) because the human eye can perceive considerably more colors than levels of grey, and (b) one can extend the scale over a far greater dynamic range(s); most monitors only display 8-bit levels of grey (24-bit color) but data are often 32-bit or more in dynamic range.
The topic of visual perception of data is a fascinating one indeed and has been addressed in several treatises over the years. However in my experience, I have found the most (subjectively) pleasing results to come from visual artists (painters) who seem to have a natural instinct for such representations. A visit to any good art museum should convince most people of this!
Sorry if this is a rather brief and simplistic answer to your question but maybe it helps some!
Peter
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
the most common use of Pseudo color is to enhance the contrast of the images and make small details more visible.
A little background:
Computer monitors are normally set to "True Color". On most graphics cards that means 32 bit of information per pixel, or "Millions of colors" as they say. However, each pixel is represented by 3 colors (Red, Green, and Blue), and each of these colors can take on an 8 bit value. 8 bits mean, that there are 256 shades of each color available, which can be combined to give you the "millions of colors" (256 x 256 x 256). What is not so obvious, that for gray levels you need to combine the 3 colors in at the same strength, i.e. black is 0,0,0, medium gray is 128,128,128 and white is 255,255,255. This shows, that even if your monitor can display millions of colors, it can usually only show 256 levels of gray. Take into account, that the human eye can distinguish perhaps 50 or so levels of gray and modern cameras can provide anywhere from 4000 to 64,000 levels of gray, and the need for different color schemes becomes obvious.
Enter the pseudo colors.
There are as many pseudo color schemes as you can think of. Several have become "standards". Among them definitely the "black body" scheme, and the "spectrum" color scheme. The "black body" is perhaps more intuitive, as it basically goes from Red to White. This provides a linear scale, which is easy to understand to anybody who has seen a metal heated (and perhaps burned himself or herself), and it is probably easier to discern small contrasts both in the red and the bright parts of the spectrum than in b/w images. However, it does not make full use of the capabilities of a monitor. The "spectrum" pseudo color, on the other hand, makes full use of the availble color spectrum, perhaps at the price of an intuitive understanding. It may be better suited to images that have "many" gray levels, which all need to be discerned. If used wrongly, however, the spectrum pseudo color can also lead to misleading coloring.
I hope this helps.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Alwyn Eades [mailto:jae5-at-lehigh.edu] Sent: Friday, January 02, 2004 10:28 To: MSA listserver
In the latest issue of Microscopy Today there is an article on pseudo-coloring of images. Since this is not something that I normally do, I read the article with interest. I was surprised to find that the author says that biologists nearly always use the "spectrum" color table for the pseudo-coloring of grayscale images. I had thought that those who study visual perception had found that among pseudo-color scales the "thermal" scale is much better than all the others at providing a good intuitive reading of the image. This is, I think, the scale that Photoshop calls "Black Body". Can someone please clear up my confusion. -- ......... 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 Jan 3 07:27:59 2004
} ... } ... In the STEM/X-Ray/EELS biological microanalysis field } most of us use some variation of the "black Body" scale } which of course more closely parallels the greyscale that } is intuitively quantitative anyway (black = 0, shades of } grey through white represent more positive values).
I remember an M&M '99 session, which introduced a pseudo-color scale for quantitative images (e.g., elemental distributions, maps). That is, ranges of color for representing "orders of magnitude" ... or ranges we might refer to as "major", "minor", "trace", or "undetected". The session was intended to be its introduction, such that its color ranges would become familiar to, and used by all, as so that quantitative images could be actually compared. I thought it was interesting concept at the time, but also felt it needed some refinement. Unfortunately my inadequate notetaking didn't allow for me to ever find the color table and download it.
I have no idea if it was mentioned in the MT article, but the people who had introduced the color scheme were from NIH (or, was it NIST?). I believe it is too bad the color scheme never did rise to common use. That is, even if I did feel it still needed some refinement, it would be a good thing if quantitative images could all be compared.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 3 10:27:30 2004
} In Dec, 2001, a group of animal rights protestors in Cambridge announced that } they intended to sue "Christianity as a whole" and anyone who celebrates } Christmas. The shock announcement comes after years of protesting against } Christmas which, they say, causes unnecessary cruelty to turkeys.
And ignore the use of reindeer as beasts of burden? Or use of swine as ham? Sounds pretty discriminatory to me.
Some people think the New Year is when it is because it was the celebration of Jesus's circumcision. If we think this is bad (e.g. castration ritual), do we refuse to follow the calendar? BTW, the holiday of New Year has become almost global.
This is a time of year when we should take some time off and relax. This message is apropos to this bboard specifically because for a few days I didn't think about microscopy at all. I read novels, slept, ate ham and argued with family over the Iraq war and mostly trivial stuff. This is happy holidays.
Now, can we get back to microscopy and drop the holiday stuff? Even if some of us won't be completing our Christmas celebrations until the 6th?
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 3 11:40:17 2004
Question: Hello there, I am a starter who wishes to get her grandchildren interested in a world beyond TV & computer games. I started using computers when I purchased my first 128K Mac back in 1985 . My present Mac is a G3 192MB ram & 40 GB hard drive.
I recently aquired a second hand "MOTIC Biological Series B1 223A " but I have found that I cannot use my lovely Fuji S602Z digital camera to take photos.
Do you have any ideas which will enable me to combine the use of the hardware that I possess? I feel that the hardest part is getting software that will enable me to join up to the Macintosh even if I purchased a new camera.
I would really like to take the photos digitally but is it impossible with my present configuration? i would appreciate any comments please
} Email: faj-at-highway1.com.au } Name: Faye Taylor } } Organization: Amateur } } Education: Undergraduate College } } Location: Perth, Western Australia } } Question: Hello there, } I am a starter who wishes to get her grandchildren interested in a } world beyond TV & computer games. I started using computers when I } purchased my first 128K Mac back in 1985 . My present Mac is a G3 } 192MB ram & 40 GB hard drive. } } I recently aquired a second hand } "MOTIC Biological Series B1 223A " but I have found that I cannot } use my lovely Fuji S602Z digital camera to take photos. } } Do you have any ideas which will enable me to combine the use of the } hardware that I possess? } I feel that the hardest part is getting software that will enable me } to join up to the Macintosh even if I purchased a new camera. } } I would really like to take the photos digitally but is it } impossible with my present configuration? } i would appreciate any comments please
Faye -
You don't say WHY you can't take photos with your equipment! I suggest that you contact microscopeworld.com. They sell many Motic scopes under the U.S. brand name "National", plus camera connectors, so you should be able to get specific advice on your problem.
You'll find abundant microscopy resources for the grandkids at the MICRO website; URL below.
Caroline
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 3 15:23:11 2004
Friends, here is the table of content of the last 2003 issue of J.Microscopy (OXF) on Microscopy in the Nanobioscience.
215 Foreword A. Diaspro
217 Polysaccharide properties probed with atomic force microscopy N. I. Abu-Lail, T. A. Camesano
239 Encapsulated yeast cells inside Paramecium primaurelia: a model system for protection capability of polyelectrolyte shells S. Krol, O. Cavalleri, P. Ramoino, A. Gliozzi, A. Diaspro
244 Insights into the regulation of transcription by scanning force microscopy R. T. Dame, C. Wyman, N. Goosen
254 Monitoring enzymatic reactions in nanolitre wells I. T. Young, R. Moerman, L. R. Van Den Doel, V. Iordanov, A. Kroon, H. R. C. Dietrich, G. W. K. Van Dedem, A. Bossche, B. L. Gray, L. Sarro, P. W. Verbeek, L. J. Van Vliet
264 The molecular machines of DNA repair: scanning force microscopy analysis of their architecture A. Janiijevi, D. Ristic, C. Wyman
273 TectoRNA and 'kissing-loop' RNA: atomic force microscopy of self-assembling RNA structures H. G. Hansma, E. Oroudjev, S. Baudrey, L. Jaeger
280 The nacre protein perlucin nucleates growth of calcium carbonate crystals S. Blank, M. Arnoldi, S. Khoshnavaz, L. Treccani, M. Kuntz, K. Mann, G. Grathwohl, M. Fritz
292 Atomic force microscopy study of living diatoms in ambient conditions I. C. Gebeshuber, J. H. Kindt, J. B. Thompson, Y. Del Amo, H. Stachelberger, M. A. Brzezinski, G. D. Stucky, D. E. Morse, P. K. Hansma
300 Self-assembly and recrystallization of bacterial S-layer proteins at silicon supports imaged in real time by atomic force microscopy E. S. Györvary, O. Stein, D. Pum, U. B. Sleytr
307 Fluorescent resolution target for super-resolution microscopy P. R. H. Stark, L. J. Rinko, D. N. Larson
I hope is of interest
All my best ALby
....................................................................... ................................... .. non basta, resistere, resistere...resistere ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Alberto Diaspro Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa, Italy voice: +39-0103536426/480 fax 010314218 diaspro-at-fisica.unige.it, http://www.lambs.it http://wiley.com/cda/product/0,,0471409200,00.html
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 4 17:50:49 2004
I meant the "visual perception" of quantitation which is what Alwyn's initial observation referred to. Of course I agree totally with you that its easer to distinguish different colors from one another than shades of any color or grey. The question is rather "is bright green more or less than yellow?" Any quantitative color scale must also have factored in parameters such as Hue, Saturation abd Brightness; this is one of the main failings of the "spectrum scale" - it doesn't!
Cheers etc
Peter
} Hello Peter, } } in actuality, the situation is a bit more complex than that. I am looking at } the LUT right now that we have in our analySIS software, and you are of } course right that a pixel with no intensity (intensity 0) is displayed as } black (0,0,0). However, the intensity "1" is displayed as (R:41, G:0, B:0). } Technically, it goes from black to white, but realistically, it goes from } "dark red" to white. } } The situation becomes mor complex at the other end. To make yellow, you have } to add in a green component, and to make white you also have to add in blue. } So, it is not straightforward "black to white" or "red to white". Other } colors get mixed in at the higher intensities to make yellow to white. } } As for qantitation: I am not sure, what you are referring to. Quantitaion is } best done on the b/w images with the help of a computer, which has no } problems distinguishing intensity 2976 from 2977. For the display of small } contrasts for the human eye I agree with you, but there an even better } choice is the spectrum LUT. It's easier to distinguish yellow from green } than it is to distinguish "dark orange" from "darker orange". } } mike } } } } -----Original Message----- } From: Peter Ingram [mailto:p.ingram-at-cellbio.duke.edu] } Sent: Friday, January 02, 2004 12:03 } To: Mike Bode } Subject: [Microscopy] RE: Psuedo color } } } } Actually the "black body" scale goes from black to white, albeit } through red, orange and yellow, not simply red to white - a vital } distinction when quantitation is involved! } } Peter } } } } --------------------------------------------------------------------------- } --- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology Duke University Medical Center Box 90319 DURHAM NC 27708-0319
Gary- I have been using a dual microprobe I purchased from Ernest F. Fullam Inc.:
900 Albany Shaker Rd. Latham NY 12110-1491 800-833-4024, 518-785-5533
Part #15855 dual, o-ring sealed manipulator. They had suggested that I purchase micromanipulators that had metal bellows rather than o-ring seals since I have a FE-SEM, but since all of the other seals on my specimen chamber were o-rings, including detectors that run in and out, I opted for the less expensive o-ring sealed option, and I have had no trouble with them. I cannot say if the microprobes I have will meet your positioning accuracy requirements as I am currently using fairly blunt tips. I also do not have verniers to check reproducibility. I just watch where I am placing them using the SEM. The probes are essentially the same as one might find on an electrical probe station for testing devices. The one problem I do have with them is that since I work with the probe tips near the pole piece, the probes induce a significant aberration. I do not know if the set screws holding the tips in are magnetic, making the problem worse or not. I do know that there are magnetic knurled nuts about 2" up the shaft. You may need to specify the materials you want the probe arms made out of, as well as working at longer WD/higher kV. I haven't bothered to correct this, or to move the probes further from the pole piece, as the resolution is adequate for my experiments. I have found Fullam to be very helpful and open to customizing their products for individual needs. They have a web site at: http://www.fullam.com/. Good luck.
Sincerely, Matthew Ervin, Ph.D. (301)394-0017 phone, (301)394-1559 fax MErvin-at-ARL.Army.mil
M/S: AMSRL-SE-RL US Army Research Laboratory 2800 Powder Mill Road Adelphi, MD 20783-1197
Disclaimer: The opinions and views expressed above are those of the author and do not necessarily represent those of the U.S. Army Research Laboratory or any other government agency
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Wednesday, December 31, 2003 10:11 PM To: MSA listserver
Hi all:
Has anyone seen a source of micro probes for SEM that allow electrical contact to a SEM chamber specimen? I need very precise positioning--like within 0.15u or better and 0.05u repeatability and stability.
I need two contact probes.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 09:42:19 2004
Our lab recently began using the "New Formulation" Kodak Electron Microscope Film 4489 and employed a number of tips for developing this film which we retrieved from the Listserver archive commencing Jan 2003
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 10:02:05 2004
I suggest that you check out "Polymer Microscopy", 2nd edition, by Sawyer & Grubb. I believe that they addressed this issue.
Cheers,
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Philip Oshel {peoshel-at-wisc.edu} To: Microscopy-at-sparc5.microscopy.com cc: Subject: [Microscopy] nylon SEM 12/23/03 02:55 PM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Materials micromavens,
We have a user doing SEM of nylon, with embedded bits. We'd like to chemically etch the nylon, which is something of an entertaining problem, since nylon is used to mask things against etchants. Does anyone have a recipe for something that will etch nylon and not silica? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 10:34:32 2004
Our lab recently began using the "New Formulation" Kodak Electron Microscope Film 4489 and employed a number of tips for developing this film which we retrieved from the Listserver archive commencing Jan 2003
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 10:44:59 2004
We here at the Neurotoxicology Labs at Rutgers University have an ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be dying. Coolwell went out of business years ago, and Zeiss pointed me to Lytron, Inc. as the company who bought Coolwell's stuff when it went under. I have tried to contact Lytron online about repairs or a possible replacement for this chiller, but they don't seem to be paying much attention to their email. I am going to call them directly, of course, but what I would like to know is if anyone can recommend another company or particular chiller model that would be appropriate for our EM, so that if Lytron continues to blow me off I will have other sources that I can try.
Hope you all had a happy holiday!
Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxcology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 10:52:17 2004
Many thanks to you for all your hard work over the past years. I very much appreciate the service that you provide for the microscopy/microanalysis community.
Cheers,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Nestor J. Zaluzec" {zaluzec-at-microscopy.c To: microscopy-at-ns.microscopy.com om} cc: Subject: [Microscopy] Administrivia: Archives for 2003 Now On-Line 01/01/04 10:37 AM
I have a Zeiss EM900 and my Coolwell also just bit the dust about 2months ago. I went with a Haskris Model 075 chiller, which includes Option (K), a 220V interlock as specified by LEO. I have two other Haskris chillers that have been very reliable (on a JEOL SEM and on a Philips TEM).
Here is the contact info:
Doug Wagner Haskris Co. 100 Kelly Street Elk Grove Village, IL 60007 847-956-6420, x243 (tel) 847-956-6595 (fax) doug-at-haskris.com
Good luck! Ann
++++++++++++++++++++++++++++++++++++++ Ann Hein Lehman Assistant Director, Electron Microscopy Facility Mailstop: LSC-314 Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 e. ann.lehman-at-trincoll.edu f. 860-297-2538 www.trincoll.edu/~alehman
-----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] Sent: Monday, January 05, 2004 11:54 AM To: Microscopy-at-sparc5.microscopy.com
Hi, all-
We here at the Neurotoxicology Labs at Rutgers University have an ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be dying. Coolwell went out of business years ago, and Zeiss pointed me to
Lytron, Inc. as the company who bought Coolwell's stuff when it went under. I have tried to contact Lytron online about repairs or a possible replacement for this chiller, but they don't seem to be paying much attention to their email. I am going to call them directly, of course, but what I would like to know is if anyone can recommend another
company or particular chiller model that would be appropriate for our EM, so that if Lytron continues to blow me off I will have other sources that I can try.
Hope you all had a happy holiday!
Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxcology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 11:39:32 2004
Hakris is a reasonably reliable chiller. I've had several on EM diff pumps. They also make a 115V version that I used on a Hitach S4500 FESEM since my lab had no chilled water supply. Unless strapped for cash, junk the old one. If it fails it can make a nasty mess of things. I think I spent 2K$ [American] for the one I bought 6 years ago.
Happy new year. [Can I say that?]
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Lehman, Ann [mailto:Ann.Lehman-at-trincoll.edu] Sent: Monday, January 05, 2004 12:39 PM To: Kathleen Roberts; Microscopy-at-sparc5.microscopy.com
Dear Kathleen,
I have a Zeiss EM900 and my Coolwell also just bit the dust about 2months ago. I went with a Haskris Model 075 chiller, which includes Option (K), a 220V interlock as specified by LEO. I have two other Haskris chillers that have been very reliable (on a JEOL SEM and on a Philips TEM).
Here is the contact info:
Doug Wagner Haskris Co. 100 Kelly Street Elk Grove Village, IL 60007 847-956-6420, x243 (tel) 847-956-6595 (fax) doug-at-haskris.com
Good luck! Ann
++++++++++++++++++++++++++++++++++++++ Ann Hein Lehman Assistant Director, Electron Microscopy Facility Mailstop: LSC-314 Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 e. ann.lehman-at-trincoll.edu f. 860-297-2538 www.trincoll.edu/~alehman
-----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] Sent: Monday, January 05, 2004 11:54 AM To: Microscopy-at-sparc5.microscopy.com
Hi, all-
We here at the Neurotoxicology Labs at Rutgers University have an ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be dying. Coolwell went out of business years ago, and Zeiss pointed me to
Lytron, Inc. as the company who bought Coolwell's stuff when it went under. I have tried to contact Lytron online about repairs or a possible replacement for this chiller, but they don't seem to be paying much attention to their email. I am going to call them directly, of course, but what I would like to know is if anyone can recommend another
company or particular chiller model that would be appropriate for our EM, so that if Lytron continues to blow me off I will have other sources that I can try.
Hope you all had a happy holiday!
Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxcology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:07:58 2004
Oooh, thank you! You just made things a lot easier for me. :o) I guess I should talk to LEO as well to see what options I need, unless your Zeiss and mine are similar enough?
God, how I love listservs....thank you, Nestor!
Thanks again- Kathleen Neurotoxicology Labs Rutgers University
Lehman, Ann wrote:
} Dear Kathleen, } } I have a Zeiss EM900 and my Coolwell also just bit the dust about } 2months ago. I went with a Haskris Model 075 chiller, which includes } Option (K), a 220V interlock as specified by LEO. I have two other } Haskris chillers that have been very reliable (on a JEOL SEM and on a } Philips TEM). } } Here is the contact info: } } Doug Wagner } Haskris Co. } 100 Kelly Street } Elk Grove Village, IL 60007 } 847-956-6420, x243 (tel) } 847-956-6595 (fax) } doug-at-haskris.com } } Good luck! } Ann } } ++++++++++++++++++++++++++++++++++++++ } Ann Hein Lehman } Assistant Director, Electron Microscopy Facility } Mailstop: LSC-314 } Trinity College } 300 Summit Street } Hartford, CT 06106 } v. 860-297-4289 } e. ann.lehman-at-trincoll.edu } f. 860-297-2538 } www.trincoll.edu/~alehman } } } -----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] } Sent: Monday, January 05, 2004 11:54 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: [Microscopy] chiller for Zeiss EM 10CA } } } } ------------------------------------------------------------------------ } ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:11:23 2004
Yes, Dr. Bonder is still at Rutgers-I just did a search for him on Rutgers' website. I don't know him personally, though, as he is on the Newark campus, where I have never been, and I am on the New Brunswick campus. Worlds apart... :o)
Kathleen
Pat Connelly wrote:
} Kathleen, } WE have been using a Haskris Co. water chiller (RO 75) since 1996 for } my Phillips 200 TEM. It works very well and the only complaint that I } have had is that we use a timer to shut down our ancient scope at } night and the one on the chiller keeps dying - it just stays on. } } This company was recommended to me when our previous chiller, also a } Haskris,died after 25 years or so, by a refrigeration specialist who } does some contract work here. } } Do you know if Dr. Ed Bonder is still at Rutgers? He was in EM the } last time I had heard from him but I do not know the exact department } - Biology? He was a grad student here at Penn a few decades ago! } } Pat Connelly } Research Specialist
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:21:08 2004
To all who wrote in reply to my question- } } Thank you for the information, you all have made my life much easier. :o) } } Now I can sit down with my boss and be able to give him some real } information about replacing this poor thing, instead of "I'm still } searching for a source..." } } Thanks again- } Kathleen } Neurotoxicology Labs } Rutgers University
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:26:57 2004
Ouch! That is an expensive repair...would LEO install a temp. sensor on a 'scope that is so old? My impression was that LEO didn't service Zeiss 'scopes anymore.
Thanks for the information- Kathleen
Ken Tiekotter wrote:
} Dear Kathleen, } } I just had a major life change as my Coolwell went down, was repaired, and } crashed again for the third and final time. The issue was exasperated by a } series of facilities failures. The outcome was about an $18k repair bill to } include a new Haskris chiller (~$5600.00) } } Unfortunately, the Zeiss EM 10CA does not a temperature sensor to determine } glycol temperature, but rather only flow rate. The repair included a } complete overhaul of the column because oil vapors were not condensing in } the diffusion pump and consequently went everywhere in the column. } } After almost 20 years with my beloved EM10, the hospital decided to donated } the scope and close my lab. You may want to check with Zeiss (LEO) about } installing a temperature sensor and automatic relay to shut the HV value if } the circulator temperature gets too high. } } Best wishes to you and your EM10! } Ken } } _______________________________________ } Kenneth L. Tiekotter, Adjunct Professor } The University of Portland } Department of Biology } 5000 N Willamette Blvd. } Portland, OR 97203 USA } } Director, MicroImaging Dx Center } Legacy Portland Hospitals } Legacy Holladay Park Medical Center } 1225 NE 2nd Avenue } Portland, OR 97232 USA } } Tel.: 503.413.5391 } } } } } } On 1/5/04 8:54 AM, "Kathleen Roberts" {kgrobert-at-rci.rutgers.edu} wrote: } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:40:57 2004
I will admit that I am not entirely sure what is wrong with it. All I know is that when I turn the 'scope on, it's fine for the first half hour or so...then a buzzer goes off-there is no indicator light to say what that buzzer is for on the 'scope, but my boss tells me that it's an overtemp alarm. If you look at the temp gauge on the chiller, it's reading way above the overtemp limit.
There was one occasion when a pump in the house distilled water system was dying, and when it was replaced, the 'scope stopped buzzing. Now, however, the distilled water system appears to be fine, and the 'scope is buzzing again, so I am assuming that it is the chiller.
I did get a local HVAC repair guy in (from the same company that resurrected our old cryostat), but he said that he couldn't do anything without the refrigeration and other specifications for that chiller. I managed to get a couple of diagrams from someone else on this list (his name escapes my memory for the moment, but thanks again anyway!), but the HVAC guy said that it wasn't enough. As Lytron isn't willing to help, I'm going to give up and get a new chiller, as this one is pretty old anyway.
Thanks, Kathleen Neurotoxicology Labs Rutgers University
McClintock, Joel wrote:
} Kathleen, } } I take care of a Zeiss 902 with a coolwell chiller. I have fixed this thing several times. What is the problem with it? Often the temp sensor goes haywire. I have found it is often the switch. I have replaced it for around $7. As others have recommended Haskris is my favorite. If money is tight I may be able to direct you to a used one. } } Joel McClintock } EM Specialist } U of Kentucky } 859-257-1242 } } -----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] } Sent: Mon 1/5/2004 11:54 AM } To: Microscopy-at-sparc5.microscopy.com } Cc: } Subject: [Microscopy] chiller for Zeiss EM 10CA } } } } } } ------------------------------------------------------------------------------ } 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- } } We here at the Neurotoxicology Labs at Rutgers University have an } ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be } dying. Coolwell went out of business years ago, and Zeiss pointed me to } Lytron, Inc. as the company who bought Coolwell's stuff when it went } under. I have tried to contact Lytron online about repairs or a } possible replacement for this chiller, but they don't seem to be paying } much attention to their email. I am going to call them directly, of } course, but what I would like to know is if anyone can recommend another } company or particular chiller model that would be appropriate for our } EM, so that if Lytron continues to blow me off I will have other } sources that I can try. } } Hope you all had a happy holiday! } } Thanks in advance- } Kathleen Roberts } Principal Lab Technician } Neurotoxcology Labs } Dept of Pharmacology & Toxicology } Ernest Mario School of Pharmacy } Rutgers University } 41 B Gordon Rd } Piscataway, NJ 08854 } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:47:21 2004
Gary, I have seen presentation on two micro-manipulator type systems: Zyvex - a MEMS based system (www.zyvex.com), and Klendiek (http://kleindiek.com, http://www.ascendinstruments.com/product_nanoprober.asp) a micro-motor based system distributed in USA by Ascend Instruments. They are geared to manipulate specimen inside a FIB or SEM chamber and both can be used for electrical measurements. The problem is that they are expensive (~100K). Since I have not actually used either one of them I cannot recommend them. We are thinking of a versatile solution that would allow TEM sample extraction, transfer, in DB-FIB and also allow us to perform occasional electrical measurements on powered devices.
Regards and Happy New Year to all.
Jerzy
PS: If you get other replies, please post them since I would also be interested in a less expensive system but with less flexibility. I do not have any vested interest in either company, we only had sales presentations from both distributors.
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 512 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-amd.com ******************************************************
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Wednesday, December 31, 2003 9:11 PM To: MSA listserver
Hi all:
Has anyone seen a source of micro probes for SEM that allow electrical contact to a SEM chamber specimen? I need very precise positioning--like within 0.15u or better and 0.05u repeatability and stability.
I need two contact probes.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 16:05:53 2004
Another source is Omniprobe, http://www.omniprobe.com/. Click the Products nav button. They can do electrical testing, TEM lift-out, mechanical testing, etc. I have no financial interest in the company.
jerzy.gazda-at-amd.com 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 } ------------------------------------------------------------------------------- } } Gary, } I have seen presentation on two micro-manipulator type systems: Zyvex - a MEMS based system (www.zyvex.com), and Klendiek (http://kleindiek.com, http://www.ascendinstruments.com/product_nanoprober.asp) a micro-motor based system distributed in USA by Ascend Instruments. They are geared to manipulate specimen inside a FIB or SEM chamber and both can be used for electrical measurements. The problem is that they are expensive (~100K). Since I have not actually used either one of them I cannot recommend them. We are thinking of a versatile solution that would allow TEM sample extraction, transfer, in DB-FIB and also allow us to perform occasional electrical measurements on powered devices. } } Regards and Happy New Year to all. } } Jerzy } } PS: If you get other replies, please post them since I would also be interested in a less expensive system but with less flexibility. I do not have any vested interest in either company, we only had sales presentations from both distributors. } } ****************************************************** } Jerzy Gazda, Ph.D. Advanced Micro Devices } Supervising Engineer 5204 E. Ben White Blvd. - MS 512 } PCAL - AIM Section Austin, TX 78741 } TEL: 1-800-538-8450, Ext. 51453 } jerzy.gazda-at-amd.com } ****************************************************** } } } -----Original Message----- } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Wednesday, December 31, 2003 9:11 PM } To: MSA listserver } Subject: [Microscopy] micro probes } } ------------------------------------------------------------------------------ } 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: } } Has anyone seen a source of micro probes for SEM that allow electrical } contact to a SEM chamber specimen? I need very precise } positioning--like within 0.15u or better and 0.05u repeatability and } stability. } } I need two contact probes. } } gary g.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 16:34:46 2004
Email: letitsnow-at-antelecom.net Name: Elizabeth Colvin
Organization: Homeschool
Education: 9-12th Grade High School
Location: Pearblossom, CA-L.A. county
Question: I have obtained several unstained blood smears. I have Wright's stain and methylene blue available. Can you please tell me how long to leave the stain on the slide before rinsing. And do I rinse with distilled water. Thank you.
Good question from Joel. The alarm (buzzer)you are hearing more than likely is the chiller fluid flow indicator on the EM10. The pressure must be maintained at 1.5 - 2liters per minute. The flow indicator is located on the right-hand side of the column inside the gray hinged 'door'. These is a small glass window to show where the float is in relationship to the flow of fluid: the higher the float, the greater the number of liters/minute.
On the front of the Coolwell chiller is also a flow indicator, which should be adjusted to meet the 1.5 - 2 liter flow on the microscope. Also check to see if the temperature gauge on the Coolwell remains the same or fluctuates. It could be the chiller is fine, but the pump is going out.
Ken
Kathleen Roberts wrote:
} } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
--------------------------------------- Kenneth L, Tiekotter, Adjunct Professor Dept. of Biology The University of Portland 5000 N Willamette Blvd, Portland, OR 97203 USA
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 17:16:32 2004
Dear List, I am posting the following for Dr. Jensen, the head of our cryo-EM group:
------------------------------------------------------------------------ ------------------------ I'm looking for an image processing scientist/computer programmer to help with our expanding biological electron tomography projects here at Caltech. We are currently imaging many specimens including cells, viruses, and purified protein complexes with a state-of-the-art 300kV, helium-cooled, energy-filtered, automated, FEG TEM. We are in the process now of purchasing a large new supercomputer for the structural biology groups. Duties would include applying existing programs as well as developing new software for image processing needs, handling large amounts of image data, managing processes on our supercomputer, working with students to help them solve image processing problems, and being a creative member of a growing scientific team. Minimum qualifications are a bachelor’s degree, strong programming skills, mathematical aptitude, an ability to work well with others, and enthusiasm for biology research. Graduate education or extended experience in related fields is preferred. Interested persons seeking either a post-doctoral position or a permanent staff position are encouraged to apply. Salary will be commensurate with qualifications. CalTech is located in Pasadena, California (a suburb of Los Angeles) next to the San Gabriel mountains, and offers an extraordinarily rich intellectual environment for computationally-inclined scientists, all within a sunny, affordable, diverse community that will make you want to stay. Please send CV and three letters of reference to jensen-at-caltech.edu or
Dr. Grant Jensen California Institute of Technology Biology Division, Mailcode 114-96 1200 E. California Blvd. Pasadena, CA 91125 ------------------------------------------------------------------------ --------------------------
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 Jan 5 17:40:32 2004
Just to add to what Ken said just below, if the flow alarm is in fact what you are hearing, then just check the various filters in the cooling water system, probably one in the chiller tank, probably another one on the input side to the microscope. Or, the lines may be plugged up somewhere with crud such as algae or corrosion products. After checking the filters and cleaning or replacing them, if problem persists may have to have scope and/or delivery lines flushed to clear them. I had to do that once for in an SEM's interior cooling lines.
Hope this helps! -- 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 ---------------------------------------------------------------------------- } Kathleen, } } Good question from Joel. The alarm (buzzer)you are hearing more than } likely is the chiller fluid flow indicator on the EM10. The pressure } must be maintained at 1.5 - 2liters per minute. The flow indicator is } located on the right-hand side of the column inside the gray hinged } 'door'. These is a small glass window to show where the float is in } relationship to the flow of fluid: the higher the float, the greater } the number of liters/minute. } } On the front of the Coolwell chiller is also a flow indicator, which } should be adjusted to meet the 1.5 - 2 liter flow on the microscope. } Also check to see if the temperature gauge on the Coolwell remains the } same or fluctuates. It could be the chiller is fine, but the pump is } going out. } } Ken } } Kathleen Roberts 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 } } ------------------------------------------------------------------------ } } Joel- } } } } I will admit that I am not entirely sure what is wrong with it. All I } } know is that when I turn the 'scope on, it's fine for the first half } } hour or so...then a buzzer goes off-there is no indicator light to say } } what that buzzer is for on the 'scope, but my boss tells me that it's } an } } overtemp alarm. If you look at the temp gauge on the chiller, it's } } reading way above the overtemp limit. } } } } There was one occasion when a pump in the house distilled water system } } was dying, and when it was replaced, the 'scope stopped buzzing. Now, } } however, the distilled water system appears to be fine, and the 'scope } } is buzzing again, so I am assuming that it is the chiller. } } } } I did get a local HVAC repair guy in (from the same company that } } resurrected our old cryostat), but he said that he couldn't do anything } } without the refrigeration and other specifications for that chiller. I } } managed to get a couple of diagrams from someone else on this list (his } } name escapes my memory for the moment, but thanks again anyway!), but } } the HVAC guy said that it wasn't enough. As Lytron isn't willing to } } help, I'm going to give up and get a new chiller, as this one is pretty } } old anyway. } } } } Thanks, } } Kathleen } } Neurotoxicology Labs } } Rutgers University } }
} --------------------------------------- } Kenneth L, Tiekotter, Adjunct Professor } Dept. of Biology } The University of Portland } 5000 N Willamette Blvd, } Portland, OR 97203 USA
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 17:41:59 2004
I did not want to imply that the "spectrum" scale is "perfect". There are many color scales and depending on what you want to see or show, one or the other might be better.
You bring up a good point, though: familiarity with the scale. Everybody can interpret a black and white scale, and the thermal scale is also very intuitive. Once we get to more colors, I would say that the spectrum scale is familiar to most people, red on one end, blue on the other. Other scales need more explanation or a color scale bar on the Image.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Peter Ingram [mailto:p.ingram-at-cellbio.duke.edu] Sent: Sunday, January 04, 2004 16:54 To: Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
Steven; I was kidding about being a Raelian. I am agnostic. I celebrate the spirit of Christmas, not the religious part. My wife is Turkish, and is a Moslem. She loves Christmas, as does our daughter. We are presently being visited by my wife's mother and sister. Despite the fact that they are both Moslem, and neither of them speaks English, they loved their first Christmas. When my wife celebrates her holidays, I celebrate with her. My brother's wife is Jewish (another mixed marriage-agnostic and Jewish). Their family celebrates both Hanukah and Christmas. Their two kids love having two holidays to celebrate. As for the spirit of Christmas, I am sorry to hear that it has eluded you. You can give gifts to loved ones any time, not just Christmas. You can try to bring more cheer into peoples lives any time, not just Christmas. You can get together with your family any time, not just Christmas. You can put up special decorations any time, not just Christmas, and you can listen to wonderful music any time, not just Christmas. It's just that human nature is such that we sometimes need a special stimulus to prioritize all this in a hectic world, and Christmas does just that-and more. Finally, if you have ever looked in to the eyes of a young child contemplating Santa Claus' imminent arrival, and shared that wonder, then you would know that this a wonderful thing to have. Steven, you have some homework to do. You need to find out what Christmas Spirit is. If you do it right, you will have a Happy New Year.
John Mardinly 408-765-2346 877-277-1182
-----Original Message----- } From: Steven E. Slap [mailto:siksik03-at-comcast.net] Sent: Monday, January 05, 2004 5:01 AM To: Mardinly, John; Microscopy-at-sparc5.microscopy.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 01:59:15 2004
} You bring up a good point, though: familiarity with the scale.
And one (little) cent more : I had a look a few weeks ago to the biology manual from my daughter (secondary school, french 3°, i. e. 15 years old), an all the EM and SEM pictures were in pseudo color, whith different color rules from one picture to an other and without any mention that it was "false colors" and that the true signal was a monochrome level variation ! I've than understood why a student asked my once, why we dont't have color images on the SEM ... " Not enough monney to pay it ?" asked he ! Familarity with a "false" color scale, can be an obstacle to understand the way the images was obtained (and to understand the image itself, perheps).
So pseudo color, why not of coarse, but with a color scale along a border of the picture, like the micron bar, as is often done on AFM images.
J. Faerber IPCMS Strasbourg France
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 03:36:16 2004
John I was thinking, it's only my family is "so creative". When we came to US my kids very quickly figured out what to do: ever since we do celebrate everything. Catholic Christmas, New Year, then Russian Christmas, then Russian "Old" New Year... The whole point there was to have gifts for every holiday... As far as I remember my kids also enjoyed some Jewish holidays when they had school off... I really like you description: "the spirit of Christmas". I think, good spirit should unify us, not separate by religious believe... Sergey.
At 11:48 PM 1/5/2004 -0800, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 05:30:26 2004
I'll also put in my $.02 worth (Cdn) for Haskris chillers. I've owned both Haskris and Neslab ones over the years, but what I really like about the Haskris models I've used is the fact that the water tank/reservoir has a removable lid, so you can always look inside and visually inspect the condition of the water. The Neslab one we have now, though reasonably reliable and all, has a completely sealed water tank with just a narrow little filler neck, so you can never see what's going on inside. Interestingly, both companies use the same water pumps supplied by a third company somewhere in Indiana, I believe. These pumps are rebuildable and replaceable of course, as are the electric motors that power them, so it's often possible to keep a chiller running for a very long time before it actually has to be replaced. To choose an appropriate model for your particular application you just need to know how much water (usually gallons/minute) and at what temperature your particular instrument needs it, then match that up to the model listing. There's not much point in greatly exceeding the needs of your instrument with a bigger chiller than necessary, since the motor/pump will be running constantly anyway. Some folks will let one big chiller cool several instruments. No doubt this is pretty cost-effective, but the down-side is apparent when the chiller craps out and you now have several instruments down instead of just one.......
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] Sent: Monday, January 05, 2004 12:54 PM To: Microscopy-at-sparc5.microscopy.com
Hi, all-
We here at the Neurotoxicology Labs at Rutgers University have an ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be dying. Coolwell went out of business years ago, and Zeiss pointed me to Lytron, Inc. as the company who bought Coolwell's stuff when it went under. I have tried to contact Lytron online about repairs or a possible replacement for this chiller, but they don't seem to be paying much attention to their email. I am going to call them directly, of course, but what I would like to know is if anyone can recommend another company or particular chiller model that would be appropriate for our EM, so that if Lytron continues to blow me off I will have other sources that I can try.
Hope you all had a happy holiday!
Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxcology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 05:50:44 2004
I'm searching for some decent disposable blades for our Leica Microtome to get sections of living hearts. I heard that Feather Blades should do the trick. Has anyone experience with getting sections of living cardiomyoctes and could give me an address of a supplier of blades, preferrably in Germany/Europe ?
Thanks
Michael Didié
-- +++ GMX - die erste Adresse für Mail, Message, More +++ Neu: Preissenkung für MMS und FreeMMS! http://www.gmx.net
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 07:29:15 2004
Although these are debates that should take place somewhere, this is not the forum for them. Unless one can tie a microscope to this line of remarks I would rather not see this on this listserver.
Just my opinion.
Peter Tomic Agere Systems
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, January 06, 2004 2:48 AM To: Steven E. Slap; Microscopy-at-sparc5.microscopy.com Cc: Microscopy-at-MSA.Microscopy.Com
Steven; I was kidding about being a Raelian. I am agnostic. I celebrate the spirit of Christmas, not the religious part. My wife is Turkish, and is a Moslem. She loves Christmas, as does our daughter. We are presently being visited by my wife's mother and sister. Despite the fact that they are both Moslem, and neither of them speaks English, they loved their first Christmas. When my wife celebrates her holidays, I celebrate with her. My brother's wife is Jewish (another mixed marriage-agnostic and Jewish). Their family celebrates both Hanukah and Christmas. Their two kids love having two holidays to celebrate. As for the spirit of Christmas, I am sorry to hear that it has eluded you. You can give gifts to loved ones any time, not just Christmas. You can try to bring more cheer into peoples lives any time, not just Christmas. You can get together with your family any time, not just Christmas. You can put up special decorations any time, not just Christmas, and you can listen to wonderful music any time, not just Christmas. It's just that human nature is such that we sometimes need a special stimulus to prioritize all this in a hectic world, and Christmas does just that-and more. Finally, if you have ever looked in to the eyes of a young child contemplating Santa Claus' imminent arrival, and shared that wonder, then you would know that this a wonderful thing to have. Steven, you have some homework to do. You need to find out what Christmas Spirit is. If you do it right, you will have a Happy New Year.
John Mardinly 408-765-2346 877-277-1182
-----Original Message----- } From: Steven E. Slap [mailto:siksik03-at-comcast.net] Sent: Monday, January 05, 2004 5:01 AM To: Mardinly, John; Microscopy-at-sparc5.microscopy.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:12:53 2004
Question: I am looking for procedures for various microchemical tests such as protein. Where would be a good resource for finding such procedures? Thanks you for your time,
A number of years ago I took a SEM course and I was told the following:
It seems a number of students comment to their professor that they would like a real time false color display on the SEM. At the time this was not a inexpensive request or total practical. The next day the students found a coffee mug full of Sharpie color markers next to the glass CRT screen and a note welcoming them to "Sharpie Color Technology."
Stay Safe................
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:36:44 2004
Hi All, I know we are not a theological forum ( and I'm not one, I'm just a SEM tech) I agree with the last mail about unifying us. This is only a way as we can Unify Religion with Science including Microscopy Microscopy and Science has discovered a world full of distinctive marks of intelligent design so we can not deny the chance of the existence of a creator, so if we want to give honor to him we have to make sure that we are worshipping him in a good way.
As much of us are from different cultures, we also have to be united, an example of unity is the language, in this case English. But What about religions, does exist a language in which we agree about belief? Even when some religious people have influenced wars, Yes, does exist : is the True. And I'm not being ambiguous I'm talking about the true about the date of Christ birth ( where Christmas is originated), the true about Mexican traditions, I mean all of them over the world, dates, even about the origin of the live (because religions talk about the creator). Knowing the true about our own traditions and beliefs and talk about them to others will unify us.
I agree about giving gifts, I enjoy accepting them and I deeply appreciate when people give them at any day of the year, without expectation.
Sincerely
Rafael Peña SEM Tech Process Enginering Tel 83297100 Ext 4721 Monterrey Nl Mexico
----- Forwarded by Rafael Pena/MONT3/MFG/KEMET/US on 01/06/04 08:29 AM -----
"Tomic, Peter (Peter)" {ptomic-at-agere.com} 01/06/04 07:32 AM
To: {Microscopy-at-sparc5.microscopy.com} cc: {Microscopy-at-MSA.Microscopy.Com} , (bcc: Rafael Pena/MONT3/MFG/KEMET/US) bcc: Rafael Pena/MONT3/MFG/KEMET/US Subject: [Microscopy] RE: RE: Dec. 25
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Although these are debates that should take place somewhere, this is not the forum for them. Unless one can tie a microscope to this line of remarks I would rather not see this on this listserver.
Just my opinion.
Peter Tomic Agere Systems
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, January 06, 2004 2:48 AM To: Steven E. Slap; Microscopy-at-sparc5.microscopy.com Cc: Microscopy-at-MSA.Microscopy.Com
Steven; I was kidding about being a Raelian. I am agnostic. I celebrate the spirit of Christmas, not the religious part. My wife is Turkish, and is a Moslem. She loves Christmas, as does our daughter. We are presently being visited by my wife's mother and sister. Despite the fact that they are both Moslem, and neither of them speaks English, they loved their first Christmas. When my wife celebrates her holidays, I celebrate with her. My brother's wife is Jewish (another mixed marriage-agnostic and Jewish). Their family celebrates both Hanukah and Christmas. Their two kids love having two holidays to celebrate. As for the spirit of Christmas, I am sorry to hear that it has eluded you. You can give gifts to loved ones any time, not just Christmas. You can try to bring more cheer into peoples lives any time, not just Christmas. You can get together with your family any time, not just Christmas. You can put up special decorations any time, not just Christmas, and you can listen to wonderful music any time, not just Christmas. It's just that human nature is such that we sometimes need a special stimulus to prioritize all this in a hectic world, and Christmas does just that-and more. Finally, if you have ever looked in to the eyes of a young child contemplating Santa Claus' imminent arrival, and shared that wonder, then you would know that this a wonderful thing to have. Steven, you have some homework to do. You need to find out what Christmas Spirit is. If you do it right, you will have a Happy New Year.
John Mardinly 408-765-2346 877-277-1182
-----Original Message----- } From: Steven E. Slap [mailto:siksik03-at-comcast.net] Sent: Monday, January 05, 2004 5:01 AM To: Mardinly, John; Microscopy-at-sparc5.microscopy.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:36:44 2004
Hi All, I know we are not a theological forum ( and I'm not one, I'm just a SEM tech) I agree with the last mail about unifying us. This is only a way as we can Unify Religion with Science including Microscopy Microscopy and Science has discovered a world full of distinctive marks of intelligent design so we can not deny the chance of the existence of a creator, so if we want to give honor to him we have to make sure that we are worshipping him in a good way.
As much of us are from different cultures, we also have to be united, an example of unity is the language, in this case English. But What about religions, does exist a language in which we agree about belief? Even when some religious people have influenced wars, Yes, does exist : is the True. And I'm not being ambiguous I'm talking about the true about the date of Christ birth ( where Christmas is originated), the true about Mexican traditions, I mean all of them over the world, dates, even about the origin of the live (because religions talk about the creator). Knowing the true about our own traditions and beliefs and talk about them to others will unify us.
I agree about giving gifts, I enjoy accepting them and I deeply appreciate when people give them at any day of the year, without expectation.
Sincerely
Rafael Peña SEM Tech Process Enginering Tel 83297100 Ext 4721 Monterrey Nl Mexico
----- Forwarded by Rafael Pena/MONT3/MFG/KEMET/US on 01/06/04 08:29 AM -----
"Tomic, Peter (Peter)" {ptomic-at-agere.com} 01/06/04 07:32 AM
To: {Microscopy-at-sparc5.microscopy.com} cc: {Microscopy-at-MSA.Microscopy.Com} , (bcc: Rafael Pena/MONT3/MFG/KEMET/US) bcc: Rafael Pena/MONT3/MFG/KEMET/US Subject: [Microscopy] RE: RE: Dec. 25
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Although these are debates that should take place somewhere, this is not the forum for them. Unless one can tie a microscope to this line of remarks I would rather not see this on this listserver.
Just my opinion.
Peter Tomic Agere Systems
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, January 06, 2004 2:48 AM To: Steven E. Slap; Microscopy-at-sparc5.microscopy.com Cc: Microscopy-at-MSA.Microscopy.Com
Steven; I was kidding about being a Raelian. I am agnostic. I celebrate the spirit of Christmas, not the religious part. My wife is Turkish, and is a Moslem. She loves Christmas, as does our daughter. We are presently being visited by my wife's mother and sister. Despite the fact that they are both Moslem, and neither of them speaks English, they loved their first Christmas. When my wife celebrates her holidays, I celebrate with her. My brother's wife is Jewish (another mixed marriage-agnostic and Jewish). Their family celebrates both Hanukah and Christmas. Their two kids love having two holidays to celebrate. As for the spirit of Christmas, I am sorry to hear that it has eluded you. You can give gifts to loved ones any time, not just Christmas. You can try to bring more cheer into peoples lives any time, not just Christmas. You can get together with your family any time, not just Christmas. You can put up special decorations any time, not just Christmas, and you can listen to wonderful music any time, not just Christmas. It's just that human nature is such that we sometimes need a special stimulus to prioritize all this in a hectic world, and Christmas does just that-and more. Finally, if you have ever looked in to the eyes of a young child contemplating Santa Claus' imminent arrival, and shared that wonder, then you would know that this a wonderful thing to have. Steven, you have some homework to do. You need to find out what Christmas Spirit is. If you do it right, you will have a Happy New Year.
John Mardinly 408-765-2346 877-277-1182
-----Original Message----- } From: Steven E. Slap [mailto:siksik03-at-comcast.net] Sent: Monday, January 05, 2004 5:01 AM To: Mardinly, John; Microscopy-at-sparc5.microscopy.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:49:58 2004
} The next day the students found a coffee mug full of } Sharpie color markers next to the glass CRT screen and } a note welcoming them to "Sharpie Color Technology."
Ah, yes... Interactive computer graphics.
Bruce Girrell
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:54:27 2004
To the listserver, Is there an easier way to make holey carbon (small 1.5 um) films, other than steaming formvar/evaporating carbon and dissolving formvar with solvents? If not who sells good small holed pure carbon films.
thanks mike d
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:58:33 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} I heard that Feather Blades should do the trick. Has anyone experience with } getting sections of living cardiomyoctes and could give me an address of a } supplier of blades, preferrably in Germany/Europe ? } } Thanks } } Michael Didié } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 09:40:53 2004
Handbook of Chemical Microscopy, Vol II. (1940) E. M. Chamot and C. W. Mason
(1989 Reprints available from McCrone Research Institute, Chicago, IL.)
Karl Hagglund (513) 634-0146
} From: lookerr-at-battelle.org (by way of Ask-A-Microscopist) on 01/06/2004 02:15 PM GMT
lookerr-at-battelle.org To: microscopy-at-ns.microscopy.com (by way of Cc: (bcc: Karl Hagglund-KW/PGI) Ask-A-Microscopist) Subject: [Microscopy] AskAMicroscopist: microchemical tests
01/06/2004 09:15 AM
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Email: lookerr-at-battelle.org Name: Ron Looker
Organization: Battelle
Education: Graduate College
Location: Columbus, OH
Question: I am looking for procedures for various microchemical tests such as protein. Where would be a good resource for finding such procedures? Thanks you for your time,
Michael, Assuming your message has lost a little in translation you might like to go to the Leica's own web site www.leica-microsystems.com. They their own (feather) microtome blades but are you actually using a vibratome?
I have just seen an ad in Microscopy and analysis Jan 2004 for a new live cell cutting module for their microdissection kit for live tissue cultures. (I have no commercial interest)
Gill Brown
GlaxoSmithKline Medicines Research Centre, STEVENAGE,
""Michael Didié"" {Michael.Didie-at-gmx.de}
06-Jan-2004 11:53
To: Microscopy
cc: Subject: [Microscopy] Feather Blades
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Hello everyone,
I'm searching for some decent disposable blades for our Leica Microtome to get sections of living hearts. I heard that Feather Blades should do the trick. Has anyone experience with getting sections of living cardiomyoctes and could give me an address of a supplier of blades, preferrably in Germany/Europe ?
Thanks
Michael Didié
-- +++ GMX - die erste Adresse für Mail, Message, More +++ Neu: Preissenkung für MMS und FreeMMS! http://www.gmx.net
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 10:04:41 2004
There is a thoughtful, well-presented magazine called Science & Spirit that addresses the intersection under discussion, which may be an appropriate venue for this dialogue. (It might make a good article for them if anyone wants to pursue it!)
-- *************************************************************** Do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro Journeys in Microspace (Columbia University Press, 1995) http://www.lsc.org/antarctica/front.html
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 10:10:32 2004
Well, knowing my boss, he will want to start with the smaller stuff -all the suggestions of what to check on the Coolwell that you and everyone else has been sending me (thank you oh so VERY much, everyone!)-and then when all those have been exhausted, go buy a Haskris. So, if it is not too much trouble, could you please dig up & send me that Grainger part number, just in case?
I will check everyone's suggestions and see if they work. At the very least, it would be nice to be at least somewhat functional until we get the new chiller in, assuming that the Facilities people here can figure the Coolwell out without diagrams. :o)
Muchas gracias, Kathleen Neurotoxicology Labs Rutgers University
McClintock, Joel wrote:
} Kathleen, } } This sounds real familiar. I suspect the switch is your problem with this chiller. No matter what chiller you have the compressor comes on then off, back and forth. On the front of this chiller right in the middle should be a temp adjustment, don't recall exact wording, but if you take a flat blade screw driver and move it around you should hear the cmpressor come on. If you do watch the temperature and see if it goes down. If so the compressor is working. Your problem then is something is not telling it ot come on properly. In my experience it is the switch from the thermocouple. Coolwells are really sensitive and when operating correctly keep the water temp pretty steady. THe compressor comes off and on, on and off quite often. The thermocouple initiates this. The thermocouple is stuck in the water and a copper line with a flat copper plate at it's end. This plate buts up to a switch. The switch has a little button on it and is pretty sensitive. As the water temp changes the copper swells and shrinks turning the switch off and on respectively. If the switch is sticky or fails things don't work. Doesn't take much as you might imagine the copper moves very little. I can dig up the number for this switch if you need it as I got it through a company called Grainger. Sound like you are going with a new one though. Good luck. } } One thing to keep in mind is whether you have a Haskris chiller or coolwell over time maintenance and/or repair will be necessary. Over the years I have called Haskris many times and their phone support is very very good. The only way to get any help on a Coolwell is if you had a time machine and even then I would not be hopeful. The design of Haskris is service friendly. } } Joel } } -----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] } Sent: Mon 1/5/2004 2:50 PM } To: McClintock, Joel; Microscopy-at-sparc5.microscopy.com } Cc: } Subject: [Microscopy] Re: chiller for Zeiss EM 10CA } } } } Joel- } } I will admit that I am not entirely sure what is wrong with it. All I } know is that when I turn the 'scope on, it's fine for the first half } hour or so...then a buzzer goes off-there is no indicator light to say } what that buzzer is for on the 'scope, but my boss tells me that it's an } overtemp alarm. If you look at the temp gauge on the chiller, it's } reading way above the overtemp limit. } } There was one occasion when a pump in the house distilled water system } was dying, and when it was replaced, the 'scope stopped buzzing. Now, } however, the distilled water system appears to be fine, and the 'scope } is buzzing again, so I am assuming that it is the chiller. } } I did get a local HVAC repair guy in (from the same company that } resurrected our old cryostat), but he said that he couldn't do anything } without the refrigeration and other specifications for that chiller. I } managed to get a couple of diagrams from someone else on this list (his } name escapes my memory for the moment, but thanks again anyway!), but } the HVAC guy said that it wasn't enough. As Lytron isn't willing to } help, I'm going to give up and get a new chiller, as this one is pretty } old anyway. } } Thanks, } Kathleen } Neurotoxicology Labs } Rutgers University
} } McClintock, Joel wrote: } } } Kathleen, } } } } I take care of a Zeiss 902 with a coolwell chiller. I have fixed this thing several times. What is the problem with it? Often the temp sensor goes haywire. I have found it is often the switch. I have replaced it for around $7. As others have recommended Haskris is my favorite. If money is tight I may be able to direct you to a used one. } } } } Joel McClintock } } EM Specialist } } U of Kentucky } } 859-257-1242 } } } } -----Original Message----- } } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] } } Sent: Mon 1/5/2004 11:54 AM } } To: Microscopy-at-sparc5.microscopy.com } } Cc: } } Subject: [Microscopy] chiller for Zeiss EM 10CA } } } } } } } } } } } } ------------------------------------------------------------------------------ } } 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- } } } } We here at the Neurotoxicology Labs at Rutgers University have an } } ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be } } dying. Coolwell went out of business years ago, and Zeiss pointed me to } } Lytron, Inc. as the company who bought Coolwell's stuff when it went } } under. I have tried to contact Lytron online about repairs or a } } possible replacement for this chiller, but they don't seem to be paying } } much attention to their email. I am going to call them directly, of } } course, but what I would like to know is if anyone can recommend another } } company or particular chiller model that would be appropriate for our } } EM, so that if Lytron continues to blow me off I will have other } } sources that I can try. } } } } Hope you all had a happy holiday! } } } } Thanks in advance- } } Kathleen Roberts } } Principal Lab Technician } } Neurotoxcology Labs } } Dept of Pharmacology & Toxicology } } Ernest Mario School of Pharmacy } } Rutgers University } } 41 B Gordon Rd } } Piscataway, NJ 08854 } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 11:05:49 2004
Group, FYI, I have posted the availability of a Kevex Sigma Gold EDS processor and a Kevex 4855 Digital Beam Control Interface to the surplus equipment list. Cheers, Tom
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 11:07:24 2004
Morning Elizabeth, If you have Wright's Stain you should do the following. 1. find two glass rods that will reach across a dish/bowl on which you can support a slide with smear up. 2. Wright's Stain is dissolved in methanol (or should be! - 0.5g Wright's in 100ml (~ { 3oz) - mixed by 'trituration' (grinding with the end of a round-bottom test tube) in small amounts of the alcohol until all is dissolved!). It is delivered on the horizontal dry smear slowly, with a dropper, so that a puddle of stain covers the smear (& perhaps the entire slide). Leave for 2.5 min. 3. Add water dropwise to the surface of the stain until IT forms a puddle that covers about 1/2 of the slide. Then blow gently, back-and-forth, on the surface of the water-stain until the two are mixed well. Total time should be about 4.5 min. 4. Rinse the water-stain off the slide in gently running water and stand the slide against an inverted glass to dry - with smear down. [If at home, DO ALL staining, rinsing and drying on aluminum foil. The dye will stain Formica surfaces. Removal requires another email.] When completely dry, a coverglass can be applied using appropriate care with a permanent mountant. 5. You can also view the smear directly with an oil immersion lens (that's the way it is done in pathology labs). Oil is placed directly on the smear and then differential counting is performed. Count 100 white blood cells, identifying each, and record the distributions. A normal smear will show something like the following: 1 basophil, 2 eosinophils, 38 neutrophils (each of the previous identified by cytoplasmic granules that are dark blue, bright red and the latter pink with a segmented nucleus respectively), and 59 lymphocytes (small cells with a round nucleus and a thin rim of cytoplasm). All red blood cells should be orange and without nuclei.
The theory is this. Methylene blue (base) and eosin (acid) are mixed in water (1:1) and combine to form a precipitate. The precipitate is dried and then dissolved(?) in methanol. After the dye thoroughly penetrates the cells in the smear, the water causes the precipitate to dissociate (based on mass action). The methylene blue and eosin are then simultaneously accessible to cellular constituents and are attracted according to their individual affinities. The rinse in excess water then removes all unbound dye. Applying the dyes separately requires much more work and gives much less satisfactory results. The above dyes belong to a group of blood dyes called "Romanovsky Stains".
Coverslipping. If you do not have an oil immersion lens, you can do the following so that you can view cells with a 40X dry objective. This will work though you will have to remove the coverslip and the oil to store the smear. To keep an oiled smear, absorb the excess oil with the tip of a piece of paper towel.
DO NOT use alcohol (will leach dyes!) to remove the rest. Wrap the slide once with good quality paper and NO Scotch tape!
DO NOT try to look at the cells when dry. The image will be saturated with diffraction rings that arise through the interaction of the microscope light and the curved surfaces of the cells - which are whole in a smear (remember?).
Hope this helps,
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson-at-wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:letitsnow-at-antelecom.net] Sent: Monday, January 05, 2004 5:38 PM To: microscopy-at-ns.microscopy.com
Email: letitsnow-at-antelecom.net Name: Elizabeth Colvin
Organization: Homeschool
Education: 9-12th Grade High School
Location: Pearblossom, CA-L.A. county
Question: I have obtained several unstained blood smears. I have Wright's stain and methylene blue available. Can you please tell me how long to leave the stain on the slide before rinsing. And do I rinse with distilled water. Thank you.
Please, PLEASE, can the next person to add to this thread spell PSEUDO correctly! ..please... (if it's not too much to ask...) (whimper) :-)
Mike O'Keefe
Bruce Girrell 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 next day the students found a coffee mug full of } } Sharpie color markers next to the glass CRT screen and } } a note welcoming them to "Sharpie Color Technology." } } Ah, yes... Interactive computer graphics. } } Bruce Girrell
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 12:12:03 2004
Our lab recently began using the "New Formulation" Kodak Electron Microscope Film 4489 and employed a number of tips for developing this film which we retrieved from the Listserver archive commencing Jan 2003 .
We use a manual processing method (no nitrogen-burst agitation). Although we have now managed to essentially eliminate the background fog and "mottled" appearance, the negatives appear darker (denser) than with the previous formulation.
These darker negatives can be printed satisfactorily in the darkroom. However, we digitize our images and archive them on-line. With the "New Formulation" film and denser negative we have noticed scan lines in our
digitized images. The darker the negative, the more apparent the scan lines. We are currently investigating the plate camera emulsion selector positions on our TEM with the hope that concurrent adjustments of exposure time/emulsion setting/darkroom technique will result in a 'less dense' negative.
Has anyone else run in to this problem when digitizing their "denser" negatives? Does anyone have any thoughts beyond what we are trying?
Thank you in advance for your assistance.
Catherine Powell
Catherine Powell, Ph.D. Division of Surgical Pathology Department of Laboratory Medicine, Saint John Regional Hospital P.O. Box 2100, Saint John, NB Canada E2K 4G9 phone (506) 648-7183 FAX (506) 648-6514 email powca-at-reg2.health.nb.ca
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 12:55:28 2004
We've been through the ringer with this film and have finally come to terms with it. However, during the last three changes of D-19 developer we have gotten significantly darker negatives (including the data bar, which is added by the scope independently of negative exposure in our JEOL 1200EX), for some reason we don't yet understand. We didn't change anything in the way we mixed our developer or exposed our films, and we have no reason to think the developer has been changed. We solved this in the meantime by diluting the developer, since the first batch of negatives had already been exposed and it's not a good idea to shorten already short developing times---four minutes in our case. We added about 25% more water (i.e., another quart per gallon of developer), ran a couple test negatives, then proceeded normally with regard to time/temperature/agitation. This worked well for us. Of course, changing developer dilutions can affect the tonal response of films, but in this case the results were fine. If you try this, always run your own tests, of course.
We've had no problems with "scan lines". That's puzzling. They must be coming from the scanner, itself, which maybe indicates that some of the scanning elements (sorry, I forget the terminology---are they called pixels?) aren't doing their thing and are causing lines in the digitized image.
Good luck.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Dr. Catherine Powell [mailto:POWCA-at-reg2.health.nb.ca] Sent: Tuesday, January 06, 2004 12:15 PM To: Microscopy-at-MSA.Microscopy.Com
Our lab recently began using the "New Formulation" Kodak Electron Microscope Film 4489 and employed a number of tips for developing this film which we retrieved from the Listserver archive commencing Jan 2003
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 13:19:33 2004
Regarding the denser negs, anytime you change film in any camera system you usually need to re-calibrate camera exposure and film development. Sounds like you have done that and are getting a less dense neg that can be nicely digitized.
Regarding the scan lines flaw, you didn't mention if you are using an actual negative scanner, or a flatbed scanner to digitize the negs. But in either case, you may just need to lubricate the moving parts. In my case, I use a UMAX Vista-S8 and about once a year I need to pull the glass surfaces off to get inside and lubricate the metal bar that the scanner heads move on. They just get dry, so then they don't slide smoothly and "skip" a line here and there. Use a clean lint-free cloth with a dab of a fine oil on it, or 3-in-1 oil is what I use. In fact, I just did this lube job yesterday, and it worked!
Good luck!
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
} Our lab recently began using the "New Formulation" Kodak Electron } Microscope Film 4489 and employed a number of tips for developing this } film which we retrieved from the Listserver archive commencing Jan 2003 . } } We use a manual processing method (no nitrogen-burst agitation). } Although } we have now managed to essentially eliminate the background fog and } "mottled" appearance, the negatives appear darker (denser) than with } the previous formulation. } } These darker negatives can be printed satisfactorily in the darkroom. } However, we digitize our images and archive them on-line. With the } "New Formulation" film and denser negative we have noticed scan lines in our } digitized images. The darker the negative, the more apparent the scan } lines. } We are currently investigating the plate camera emulsion selector } positions on } our TEM with the hope that concurrent adjustments of exposure } time/emulsion setting/darkroom technique will result in a 'less dense' } negative. } } Has anyone else run in to this problem when digitizing their "denser" } negatives? Does anyone have any thoughts beyond what we are trying? } } Thank you in advance for your assistance. } } Catherine Powell } Catherine Powell, Ph.D. } Division of Surgical Pathology } Department of Laboratory Medicine, Saint John Regional Hospital } P.O. Box 2100, Saint John, NB Canada E2K 4G9 } phone (506) 648-7183 FAX (506) 648-6514 } email powca-at-reg2.health.nb.ca
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 13:32:52 2004
I haven't had any issues scanning in the darker negatives. We have an Agfa Duoscan T2500 that works well enough with these newer negatives at 1000 and 2500 ppi (the two resolutions I've tried). Before the holiday break I put together a group of images (digitally scanned ones) that had both new and old negatives included with no discernable differences between them. Other than having the scanner support discontinued its been a great and reliable scanner for lecture slides and TEM negatives (if anyone has a driver for it that works with the latest Mac OS 10.3 I would be very interested in hearing about it). As you have mentioned, it would probably be worth trying a emulsion setting series to see if that improves your results. But alas I cannot say that I've tried this yet to attempt to get the old and new density similar on the negatives.
HTH,
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
} -----Original Message----- } From: Dr. Catherine Powell [mailto:POWCA-at-reg2.health.nb.ca] } Sent: Tuesday, January 06, 2004 1:15 PM } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] TEM -Need help with scanning EM film 4489 } } } } ------------------------------------------------------------------------ -- } ---- } 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 } ------------------------------------------------------------------------ -- } ----- } } Our lab recently began using the "New Formulation" Kodak Electron } Microscope Film 4489 and employed a number of tips for developing this } film } which we retrieved from the Listserver archive commencing Jan 2003 . } } We use a manual processing method (no nitrogen-burst agitation). } Although } we have now managed to essentially eliminate the background fog and } "mottled" appearance, the negatives appear darker (denser) than with } the previous formulation. } } These darker negatives can be printed satisfactorily in the darkroom. } However, we digitize our images and archive them on-line. With the } "New } Formulation" film and denser negative we have noticed scan lines in our } } digitized images. The darker the negative, the more apparent the scan } lines. } We are currently investigating the plate camera emulsion selector } positions on } our TEM with the hope that concurrent adjustments of exposure } time/emulsion setting/darkroom technique will result in a 'less dense' } negative. } } Has anyone else run in to this problem when digitizing their "denser" } negatives? Does anyone have any thoughts beyond what we are trying? } } Thank you in advance for your assistance. } } Catherine Powell } } } } Catherine Powell, Ph.D. } Division of Surgical Pathology } Department of Laboratory Medicine, Saint John Regional Hospital } P.O. Box 2100, Saint John, NB Canada E2K 4G9 } phone (506) 648-7183 FAX (506) 648-6514 } email powca-at-reg2.health.nb.ca
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 13:34:03 2004
Thanks, Mike, for pointing out the proper spelling. The correct pronunciation of ‘psuedo’ is as in ‘blue psuedo shoes.’ And there’s nothing all that astonishing about using colored markers on the screen. When we first introduced a computer (one of the original upright Macs) in the department office a couple of decades ago, to a secretary firmly set in the old ways, we had to take turns each night cleaning the whiteout correction fluid off the screen. {;-)
Anyway, to the use and abuse of pseudo color;...
It is certainly true that human vision can only distinguish a few dozen shades of grey brightness on a display screen, as compared to a few hundred colors. Note that both of these values are far less than the 256 shades of brightness or “millions†of colors that the hardware typically controls. It is also true that trying to direct someone’s attention to the “kind of darkish grey spot†is a lot less helpful than “the yellow-orange spot†in an image (but of course, human words for colors aren’t terribly consistent or widely agreed, either - look at any set of paint chips in the turquoise-teal-seagreen-etc. family). Pseudo- or false-color certainly has some valid uses. But it is also easily misunderstood, widely abused, and often hides more in the image than it reveals. And if the viewer is one of the 5-10% of men (or the very tiny percentage of women) who have defective color vision, it is inappropriate in any case.
Firstly, of course, the table must be shown along with a numerical scale that translates it. But even then, simple spectrum lookup table is rarely if ever a good choice. The problem is that the table is typically constructed with uniform steps in hue, going around the color wheel. But human vision is notably insensitive to changes in hue in the green part of the spectrum, and much more so at the red and blue ends and through the red-to-blue purples. A perceptually uniform hue scale (which I have never seen used) would stretch these out and compress the greens and could probably produce more than a hundred discernible colors.
More colors could be seen if they were NOT fully saturated. Changing saturation and hue in a spiral pattern, or also altering brightness along with hue and saturation, can produce color tables that varied in a gradual way and produced greater ability to distinguish changes. The gradual part is important - if the colors jump around too much in discontinuous ways, the image is badly broken up (camouflaged, in effect) and the overall sense of structure, the gestalt of the image, is hidden. To some extent this happens even with a good, gradual table. The use of the “heat†or “thermo†scale is an example of a gradual and visually attractive scale, which does not break up the content of the image. But it does not actually add very much to the ability to visually distinguish small changes - perhaps a 20-40% improvement over straight grey scale (which is why color tints are also used in photographic printing, to gain the same increase). Note that the brightness increases monotonically in this scale, and that it is by contributing more steps at the dark end that the increase is obtained.
For selected purposes, carefully constructed color scales can be useful to help the viewer perceive subtle differences or make comparisons from one part of an image to another. But they need to be documented, and in most cases it is also important to show the original data as well in case the color scale can produce misinterpretation or hide other information.
It has been my experience that people are not generally assisted very much by pseudo color scales, as compared to other ways to reveal subtle detail. One of the best of these is to render the surface with elevation representing the original grey scale value. We have millions of years of evolution in our brain wiring that knows how to interpret surface images, in terms of shape and roughness. Using computer graphics to generate properly rendered images with correct perspective, and adjustable viewpoints, surface characteristics, and illumination is easy with current technology and communicates very effectively. The AFM folks use this trick too, along with color scales, although in most cases in only limited ways.
John Russ (see www.DrJohnRuss.com for a schedule of upcoming workshops on image analysis)
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 13:50:49 2004
The recent thread about the use and abuse of pseudo color is only one of many issues that have to do with an important topic that underlies just about everything that we as microscopists do - namely, we LOOK AT images. But while we are typically very concerned with the performance and specifications of our microscopes, we take for granted the performance of our visual systems, to our peril.
Over the past 5 years or so I have been invited several times to give a talk on human vision and how it impacts what microscopists see (and fail to see) in images. At the repeated urging of many people, I’ve prepared the lecture in written form. Anyone who wants to read it can download the "Seeing the Scientific Image" pdf file from my website (www.DrJohnRuss.com). If someone thinks there is a logical place to publish it (too long for Microscopy Today, and not a research paper for Microscopy and Microanalysis) please let me know.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 14:12:58 2004
Silly me, in all my crabbing about this chiller, I didn't give the model? The model # is SE-075W CZ. I got some diagrams from Eckhart Dorneich, and a manual is on the way from Joel McClintock (thank you both!), but I'll take yours too, if they match my chiller. I figure you can never have too much information, and it's always good, in this lab, to have extra copies. :o)
Thank you very much- Kathleen Neurotoxicology Labs Rutgers University
Tina Carvalho wrote:
} Hi- } } What model chiller do you have? I might have the wiring and plumbing } diagrams. } } I have a 10/A and have struggled to keep the chiller going. The scope was } down for a couple of years - oil and then water in the column! - while we } used our new LEO 912 EFTEM. The expensive new instrument can be very } frustrating, and was down once for nearly 5 months! So we got the 10/A } going again, including the chiller. We all had forgotten what a gem it is, } and right now I'd sell the new one and keep the old if I had a } choice. } } We have Hakris chillers on our other two instruments, and they are } fine. We got the very first one that Haskris built for Zeiss/LEO, and they } had a fit trying to include both a pressure and flow gauge plus that } interlock that keeps it going to cool down the diff pump after you turn } off the scope, but they are quite used to it, now. They're pretty reliable } and, fortunately, not prone to that temperature sensor screwing up like } the Coolwell. } } Over the years I've found out there are a whole lot of closet Zeiss 10s } out there - the most reliable backup even if you have a fancy new } instrument! Keep it going. My next trick is to outfit it with a digital } camera. } } Aloha, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 15:03:50 2004
Dear Catherine, I found the new formulation film was more sensitive and adjusted my auto settings in the TEM to compensate. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "Dr. Catherine Powell" {POWCA-at-reg2.health.nb.ca} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Tuesday, January 06, 2004 10:14 AM
Scanning very dense negatives can cause problems related to the scanner if your scanner is using a CCD rather than a PMT. With a CCD you can not crank up the current to boost the signal like you can with a PMT. The result is that there is not enough light i.e. signal getting to the camera array. If that is the case then you must either slow down the scan speed so that the camera can collect enough signal or if this is impossible make your negatives less dense. Our ZI Photoscan 2000 has problems with negatives above an OD of 2.3 or so. Above this lines like what you described appear. I hope this helps. Good luck.
Bob Grassucci
At 02:14 PM 1/6/04 -0400, Dr. Catherine Powell wrote:
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Robert Grassucci Howard Hughes Medical Institute at The Wadsworth Center Empire State Plaza Albany, NY 12201-0509 Phone: (518)474-5821 Fax: (518)486-2191 E-Mail: bobg-at-wadsworth.org
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 17:31:44 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (alchung-at-ucla.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, January 6, 2004 at 16:51:57 ---------------------------------------------------------------------------
Email: alchung-at-ucla.edu Name: Albert Chung
Organization: UCLA
Education: Graduate College
Location: Los Angeles, CA USA
Question: I am having issues with the silicon monoxide film tearing in the TEM. I have called Ted Pella about this problem and they are making a new batch as we speak, but I have about 50 silicon monoxide films on copper grids for which I cannot use. Any suggestions on how to approach this problem is greatly appreciated. I am operating the JEOL 100CX and the typical current saturation is about 80 mA.
If you need additional information, please let me know.
There is another one: you may bombard thin nylon film by some heavy ions (argon may be good) in the accelerator. It will produce very uniform holes. The size of holes strictly dependent from the energy and type of particle used. But, still: you need to eveporate carbon over and dissolve nylon (have no idea how to do so). As a matter of fact this technology widely used to produce filters for ultrafiltration.
Sergey
At 07:03 AM 1/6/2004, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Albert Chung wrote: ======================================================================== Question: I am having issues with the silicon monoxide film tearing in the TEM. I have called Ted Pella about this problem and they are making a new batch as we speak, but I have about 50 silicon monoxide films on copper grids for which I cannot use. Any suggestions on how to approach this problem is greatly appreciated. I am operating the JEOL 100CX and the typical current saturation is about 80 mA. ======================================================================== Support films of SiOx can be too thick or too thin or could have other features about them that render them unstable in the electron beam. This is of course exactly the same for carbon coated grids. That is why it is imperative to have in-house, as part of the grid coating process, a TEM for batch inspection purposes so that the customer does not end up being responsible for their own QC. And does not end up preparing a large number of grids only to find them unusable.
In the case of carbon replica films that were prepared but which are too thin, we have in the past, been able to resurrect them by applying another coating of carbon to strengthen them and to make grids that were once unstable, now stable. We have never done this with SiOx filmed grids but in theory, it might work the same way.
Disclaimer: SPI Supplies produces SiOx firmed grids as a part of our business and to our knowledge, we have not had any problems with film stability in the electron beam. On occasion, we do make a batch that flunks our own in-house TEM inspection process but those grids never make it into the hands of customers.
Chuck
PS: Remember that we are 100% paperless and the only way we can keep track of this kind of correspondence is if you reply using the reply feature of your e-mail software. That way the entire string of correspondence will 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 Wed Jan 7 00:09:32 2004
by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id i0769WI9018789 for {LL1-at-ns.microscopy.com} ; Wed, 7 Jan 2004 00:09:32 -0600 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id i0769WFG018785 for LL1; Wed, 7 Jan 2004 00:09:32 -0600 X-Authentication-Warning: ns.microscopy.com: mail set sender to MicroscopyL-request-at-ns.microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id i0768pI9018755 for "MListServerFilteredEmail_6-at-microscopy.com"; Wed, 7 Jan 2004 00:09:11 -0600 Received: from caduceus.jf.intel.com (fmr06.intel.com [134.134.136.7]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id i0768oI9018752 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 7 Jan 2004 00:08:50 -0600 Received: from talaria.jf.intel.com (talaria.jf.intel.com [10.7.209.7]) by caduceus.jf.intel.com (8.12.9-20030918-01/8.12.9/d: major-outer.mc,v 1.12 2003/12/18 18:58:11 root Exp $) with ESMTP id i076DUa5019496; Wed, 7 Jan 2004 06:13:30 GMT Received: from orsmsxvs040.jf.intel.com (orsmsxvs040.jf.intel.com [192.168.65.206]) by talaria.jf.intel.com (8.12.9-20030918-01/8.12.9/d: major-inner.mc,v 1.7 2003/12/18 18:58:10 root Exp $) with SMTP id i076A5Zv017386; Wed, 7 Jan 2004 06:10:20 GMT Received: from orsmsx332.amr.corp.intel.com ([192.168.65.60]) by orsmsxvs040.jf.intel.com (SAVSMTP 3.1.2.35) with SMTP id M2004010622122332399 ; Tue, 06 Jan 2004 22:12:23 -0800 Received: from orsmsx312.amr.corp.intel.com ([192.168.65.62]) by orsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(5.0.2195.5329); Tue, 6 Jan 2004 22:12:23 -0800 Received: from scsmsx402.amr.corp.intel.com ([10.3.90.16]) by orsmsx312.amr.corp.intel.com with Microsoft SMTPSVC(5.0.2195.5329); Tue, 6 Jan 2004 22:12:23 -0800 Received: from scsmsx403.amr.corp.intel.com ([10.3.90.18]) by scsmsx402.amr.corp.intel.com with Microsoft SMTPSVC(5.0.2195.5329); Tue, 6 Jan 2004 22:12:22 -0800 content-class: urn:content-classes:message MIME-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" X-MimeOLE: Produced By Microsoft Exchange V6.0.6487.1
Catherine; One of my favorite web sites, Imaging Resources http://www.imaging-resource.com has reviewed numerous film scanners. Scanning extremely dense film has always been an acid test for a scanner. The following link is to a negative scanned with a Minolta Dimage Scan Elite II Film & Slide Scanner that failed the acid test in some dark regions, and the effect there seems similar to what you describe. The site tests many other scanners with the same negatives, and many do not exhibit this artifact. http://www.imaging-resource.com/SCAN/DSEII/DSETRAA602PS.HTM
John Mardinly Intel
-----Original Message----- } From: Dr. Catherine Powell [mailto:POWCA-at-reg2.health.nb.ca] Sent: Tuesday, January 06, 2004 10:15 AM To: Microscopy-at-MSA.Microscopy.Com
Our lab recently began using the "New Formulation" Kodak Electron Microscope Film 4489 and employed a number of tips for developing this film which we retrieved from the Listserver archive commencing Jan 2003 .
We use a manual processing method (no nitrogen-burst agitation). Although we have now managed to essentially eliminate the background fog and "mottled" appearance, the negatives appear darker (denser) than with the previous formulation.
These darker negatives can be printed satisfactorily in the darkroom. However, we digitize our images and archive them on-line. With the "New Formulation" film and denser negative we have noticed scan lines in our
digitized images. The darker the negative, the more apparent the scan lines. We are currently investigating the plate camera emulsion selector positions on our TEM with the hope that concurrent adjustments of exposure time/emulsion setting/darkroom technique will result in a 'less dense' negative.
Has anyone else run in to this problem when digitizing their "denser" negatives? Does anyone have any thoughts beyond what we are trying?
Thank you in advance for your assistance.
Catherine Powell
Catherine Powell, Ph.D. Division of Surgical Pathology Department of Laboratory Medicine, Saint John Regional Hospital P.O. Box 2100, Saint John, NB Canada E2K 4G9 phone (506) 648-7183 FAX (506) 648-6514 email powca-at-reg2.health.nb.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 06:34:24 2004
We have too 2 Kevex Delta avavaible, one complete, with detector (10mm2, 138 eV, warm, but was working in July)), the other only the electronic, for spares, with 8" and 5"1/4 Bernouilli.
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
If anyone has a mechanical lab balance they would like to dispose of, I could use one. I collect/repair antique phonographs as a hobby and would like to be able to true up governor weights from time to time. I've looked at E-bay but I am leery since, if a balance moves up and down, the general public thinks it works. Some units offered are missing parts. etc.
Thanks,
Ron
-----Original Message----- } From: qualityimages [mailto:qualityimages-at-netrax.net] Sent: Wednesday, January 07, 2004 7:37 AM To: Microscopy
Posting this for a friend.
Ken Converse owner Quality Images third party SEM service Delta, PA
Hi All,
I am doing some housecleaning & have the following EDS Systems free to anyone:
As many of you may know I run a training organisation that travels world wide spreading the word on SEM, TEM and EDS operation. I also have a deep interest in "Quality in Electron Microscopy"as those who picked up our paper last year would know?
Now to the point. Once again I am picking up respected journals and finding examples of what I would call poor microscopy, but in truth it also demonstrates poor quality control!
To bring one image to mind. The micrograph is of a structure which is described as being an example of a smooth surface on a biological material. But, the micrograph was taken at 20kV, where the vast majority of the information will have come from beneath the surface softening the true surface detail.
First to remove the training aspect . Operators of SEM should be taught that by manipulation of kV and working distance one may subdue or enhance surface features. To use more than 10kV on most biological samples is asking for sub surface detail, ignore this and comments on surface irregularities are null and void in my mind. (I have to say I would probably try to use 2 to 5kV if the microscope used was produced in the last 15 years!)
Now the quality aspect. By the time a paper is published a number of steps should have been taken. Working backwards, the publisher should have the paper vetted by knowledgeable scientists who would be able to pick out the problems that I see and have them corrected prior to going to print. Next back in the chain is the laboratory that was involved with the scientist; did they check the quality of the work leaving their EM unit? Stepping back again did the scientist take the micrographs or did they receive help? Either way the training of staff and operators should overcome this type of problem! But if the results the staff and visiting operators produce are not assessed how do you know that their training is inadequate?
As the pressure to perform increases and funding decreases only the cream of our laboratories will remain. In industry there is no question about following rigid "Quality" procedures and it is not too far off that this will hit the world's EM units too! I know that this is my baby but is it not about time that we woke up to these facts?
There is an area where I believe we have room for discussion; what do you think?
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob +44 (0) 7711 606967 www.emcourses.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 10:49:04 2004
Steve, Your post (whose spirit I agree with totally) prompted a question. If I have a biological sample sputter coated with metal, isn't all the contrast coming from the metal? Does the underlying carbon based material scatter anything much? If the carbon isn't scattering much then wouldn't the problem you used for illustration matter only at high mags where distances on the order of the coat thickness are being resolved?
As ever, Tobias
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Steve, I agree entirely with you in that training (perhaps educating is a better word) is key to good and reliable results. The example you sited happens constantly. I take great pains to not only lecture about, but prove through lab exercises, the effects that varying microscope parameters have on the final image.
Unfortunately many "trained" people ask to use our facility and are denied because their training was inadequate. They are either retrained so they have the theory to go along with twiddling the knobs or rely on our "service" option (trained staff does the actual imaging). The reputation of our facility is very important.
We cannot guarantee to get perfect results with every research system on the first try but we do our best and learn from our mistakes. At least if we understand our instruments we can concentrate on sample prep to get the best possible final results...not what the investigator thinks is there but what is ACTUALLY there.
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 1/7/04 9:42 AM, "Steve Chapman" {protrain-at-emcourses.com} 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 Listers } } As many of you may know I run a training organisation that travels world } wide spreading the word on SEM, TEM and EDS operation. I also have a deep } interest in "Quality in Electron Microscopy"as those who picked up our paper } last year would know? } } Now to the point. Once again I am picking up respected journals and finding } examples of what I would call poor microscopy, but in truth it also } demonstrates poor quality control! } } To bring one image to mind. The micrograph is of a structure which is } described as being an example of a smooth surface on a biological material. } But, the micrograph was taken at 20kV, where the vast majority of the } information will have come from beneath the surface softening the true } surface detail. } } First to remove the training aspect . Operators of SEM should be taught that } by manipulation of kV and working distance one may subdue or enhance surface } features. To use more than 10kV on most biological samples is asking for } sub surface detail, ignore this and comments on surface irregularities are } null and void in my mind. (I have to say I would probably try to use 2 to } 5kV if the microscope used was produced in the last 15 years!) } } Now the quality aspect. By the time a paper is published a number of steps } should have been taken. Working backwards, the publisher should have the } paper vetted by knowledgeable scientists who would be able to pick out the } problems that I see and have them corrected prior to going to print. Next } back in the chain is the laboratory that was involved with the scientist; } did they check the quality of the work leaving their EM unit? Stepping back } again did the scientist take the micrographs or did they receive help? } Either way the training of staff and operators should overcome this type of } problem! But if the results the staff and visiting operators produce are } not assessed how do you know that their training is inadequate? } } As the pressure to perform increases and funding decreases only the cream of } our laboratories will remain. In industry there is no question about } following rigid "Quality" procedures and it is not too far off that this } will hit the world's EM units too! I know that this is my baby but is it } not about time that we woke up to these facts? } } There is an area where I believe we have room for discussion; what do you } think? } } Steve Chapman } Senior Consultant Protrain } Electron Microscopy Training and Consultancy World Wide } Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob +44 (0) 7711 606967 } www.emcourses.com } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 14:28:28 2004
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Put into TEM at low mag but still with an objective aperture in place, and 100 kV. Really spread the beam and turn up slowly. You may find that by gradually increasing the electron dose, you'll "harden" the film. This approach certainly works with formvar films and biological resin sections but I've never tried it with silicon monoxide, so I can't guarantee it'll work. -- Larry Stoter NOTE - any message other than plain text will be automatically deleted :-)
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 15:12:05 2004
Does anyone have a circuit diagram for an oldish Leitz microscope lamp power supply Type 301-314.001?
It's a beige box with an analog voltmeter, a mains switch, and a brighness adjust knob on the front panel.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 15:24:34 2004
Question: we used a spurr's resin prep on a sample of nematodes, after staining and heat-fixing the sections, we see dark ribbon-like structures on several specimens. Are these artifacts caused by air bubbles and can they be avoided? thank you, jamie
We believe the problem may have been caused by weak film. Silicon monoxide film should be as fresh as possible. Some of our customers have requested thicker coatings but this doesn't help. We make our film up the day it is requested by the customer to insure this problem doesn't occur. We suggest you purchase the minimum amount required at one time because new batches can be made up in a day or so.
John Arnott Disclaimer: Ladd Research produces a wide variety of EM supplies including substrates, such as silicon monoxide.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Larry Stoter" {larry-at-cymru.freewire.co.uk} To: "by way of Ask-A-Microscopist" {alchung-at-ucla.edu} ; {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, January 07, 2004 3:27 PM
Steve, et al.:
I agree there is a definte problem in terms of (some of) the microscopy work being used and published. Steve hits a very good case where failure to understand the technique can easy lead to poor data. (by microscopy I mean in the broadest sense: SEM, TEM, LM, SPM, etc. . . Anyone dealing with molecular biolgists labling molecules and wanting to use a confocal microscope already knowing exactly what kind of "picture" they want to get knows what I mean)
One of the really frightening aspects is that very very few people wish to learn microscopy - become microscopists - today. I have been watching this trend for several years now where users (students and faculty in my case) simple want images. They want to push a button out comes an image - that's all. If they could have the microscope automatically load the sample that would be even better. Now granted there are times when a simple click image will sufice - but more and more often researchers are failing to realize how far they are trying to push the capabilities of a microscopy technique. I have been told specifically they do NOT want to learn the microscope they want an image. And then you try and turn around and tell them the data they are collecting is bad science?
} Next back in the chain is the laboratory that was involved with the } scientist; did they check the quality of the work leaving their EM unit? } Stepping back again did the scientist take the micrographs or did they } receive help? Either way the training of staff and operators should overcome } this type of problem! But if the results the staff and visiting operators } produce are not assessed how do you know that their training is inadequate? }
There is a time you can attempt to argue with the "users" over scienfic quality, but running and EM Lab you can not dictate it - certainly not in academia, and not even in industry - yes, you can be asked for an evaluation of the data (and that is what peer review also does) but no one can really control what the data is used for after it leaves the lab.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 16:48:19 2004
We are having one of those debates that we microscopists seem to obsess about. The question is whether to store our saturated uranyl acetate solution (in dH2O) at room temp or at 4 C. Opinions, especially those backed by data, would be welcome. Happy New Year. Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
With huge headache I've established procedure in my Lab that everyone, who uses equipment in the Lab should go through mandatory training. Without this training, person just could not enter the facility (digital lock). My training course includes intense training on data collection, interpretation and sample preparation. So, it does not insure from the "bad science", but at least I felt people knows what they doing. It means, if they will present "bad data" I know, they did it on purpose... My course is about week long (2h/day) and people's reaction are very different. Nevertheless, I noticed that majority of the users finally enjoyed "good electron microscopy", because it save them time and the quality of their images is good (and confidence is great). In general, I do agree: people becomes more and more "lazy" - they want to have results doing nothing (best scenario - machine will do). It's very pity and made me very skeptical on quality of many data published. Sergey
P.S. Knowledge is power.
At 02:20 PM 1/7/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
More important - to store in the dark! At +4oC you will have lower concentration because of solubility. I prefer to store most of the chemical solutions at cold temperature. Sergey
At 02:49 PM 1/7/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
If it makes all of you feel any better, the phenomenon of users not wanting to learn how to use something, just wanting the results, is not limited to microscopy, and is a dangerous trend. This trend is ironically aided by the very advancing technologies that make truly understanding the theories and principles of what is happening more important than ever.
Too many devices, instruments and other systems have become "too easy to use". The advances in human interfaces, automation, computer controls, etc. have made it very easy for just about anyone to get results. The danger is that there is no way for someone who doesn't truly understand what's going on to know if the results are meaningful or not. "I got the answer, and it's what I was expecting, so it must be right!"
The problem is also not limited to "sophisticated" technology. We have users who no longer know that there is a darkness adjustment on a copy machine, much less how to use it. Not that long ago, you couldn't get two copies in a row to turn out without tweaking the adjustment. If someone does take the thing off "auto" and sets it dark, everyone else thinks the thing is broken...
If someone finds a "real" cure for this phenomenon, please share it with the world, because the problem is pretty universal.
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 03:26:44 2004
I fully agree with this fact that the problem is universal, and it is not only limited to microscopy. However, before one finds the "cure for this phenomenon", one should look for the cause of this problem.
If the new generation, i.e. the younger students and others, "are just wanting results", is it not the consequence of the ATTITUDE OF THE LARGE MAJORITY of those who trained them?
Those who trained the last two generations are now collecting the results of what they have done. The biggest reference for this generation is having rapid success, which means gaining large amounts of money and if possible a quick fame.
No university professor considers the students for their interest for science and their will to do something for the sake of humanity. They rank them on their good marks, without worrying about how they obtained it. And even if they do so, it is to REPRIMAND the student and not for TEACHING them the importance of honesty. A large percentage of students learn just to obtain a good mark, without thinking about why's and how's. And those few who do, are not appreciated by the professors, may be since they need more time that is not available.
A professorship is the art of teaching many things including highest human values to the students, I can not name many who have done that, or even have thought to do it. If they do not reflect such thoughts, at least they should avoid to discourage the very few who do have such values (and have truly come to learn something that can be helpful for the society) by treating them as naives and dealing with irony with them.
Some students tell me that the science professors are so arrogant that the students have no more appeal for scientific branches. } From my discussions with some undergraduate science students, their feeling is that if they go for science in their graduate school they could ruin their future and they will loose the chances of success!
I agree that the danger is real and from my understanding the solution to the problem is to reverse the trend from now on by changing the teaching philosophy. However, one should have the certitude of the importance of such a training to be able to have an impact on one's student.
Sousan Abolhassani, Ph.D. Laboratory for Materials Behaviour Paul Scherrer Institut 5232 Villigen PSI Switzerland
"John W. Raffensperger, Jr." 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 } ------------------------------------------------------------------------------- } } If it makes all of you feel any better, the phenomenon of users not } wanting to learn how to use something, just wanting the results, is not } limited to microscopy, and is a dangerous trend. This trend is } ironically aided by the very advancing technologies that make truly } understanding the theories and principles of what is happening more } important than ever. } } Too many devices, instruments and other systems have become "too easy to } use". The advances in human interfaces, automation, computer controls, } etc. have made it very easy for just about anyone to get results. The } danger is that there is no way for someone who doesn't truly understand } what's going on to know if the results are meaningful or not. "I got } the answer, and it's what I was expecting, so it must be right!" } } The problem is also not limited to "sophisticated" technology. We have } users who no longer know that there is a darkness adjustment on a copy } machine, much less how to use it. Not that long ago, you couldn't get } two copies in a row to turn out without tweaking the adjustment. If } someone does take the thing off "auto" and sets it dark, everyone else } thinks the thing is broken... } } If someone finds a "real" cure for this phenomenon, please share it with } the world, because the problem is pretty universal. } } John W. Raffensperger, Jr. } IS Manager } Helwig Carbon Products, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 04:32:18 2004
Our old camera has to be replaced and I am offered a new one for 1200 euros. This may not be too expensive but I wonder if I can get the same results with a cheaper webcam attached to the microscope. I understand it is just a question of removing the camera objective lense or to make it focus infinity.
My question is: can I expect a really better performance from a more expensive, purpose-built camera? Quite likely the camera will not be the limiting factor in the quality of micrographs, but other factors in the microscope and the preparation...
And, which factors should I cosider in the camera? I am thinking of sensitivity to poor light, gain, and so.
Thanks for all your comments,
Antonio D. Molina-García Inst. del Frio (CSIC) Madrid, Spain
PD. My main purpose for video recording from a microscope is to study ice crystal evolution during growth and recrystallization. Image is so, not too sharp ever, as the contrast between ice crystals is small. Also the sample thickness is larger than when using other sample types and, when the size of crystals is small, it is even difficult to get any light through...
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 04:54:16 2004
Second Announcement From Microscopy to Nanoscopy, Genoa, 9-11 February 2004. The 5th Practical and Intensive Workshop on 3D Confocal Microscopy will be held in Genoa, at LAMBS, Department of Physics, Via Dodecaneso 33, 16146 Genoa. The aim of the workshop, as usual, is to introduce researchers to Confocal Microscopy and related methods. This year we would like to show how it is possible to move from Microscopy to Nanoscopy, according to a Stefan Hell’s recent paper (Natur.Biotech., 21 (11), 2003, p.1347). 2004 Program includes: lessons on basic aspects of fluorescence and confocal microscopy (February 9th, 10 a.m.- 1 p.m.), lectures on applications of confocal microscopy and related methods (February 9th, 2 p.m.-7 p.m. with coffe break), experimental sessions on confocal and multiphoton architectures, experimental sessions with image analysis, processing, visualization and deconvolution software, roundtables with speakers (February 10th-11th, 8,30 a.m. - 7 p.m., including lunch break). Lecturers: Tony Wilson, Erik Manders, Alberto Diaspro, Mario Faretta, Cesare Usai. The workshop is limited to 20 students served on first-in-first-out basis. Only Monday afternoon lectures are open. The workshop fee is 250 Euros. Some grants will be available on the basis of real need. Please register or request information sending an e-mail to diaspro-at-fisica.unige.it using "confocal 5 - genoa" as subject. Please, specify if you want to attend the Workshop or only Monday afternoon lectures. Logistic help can be provided upon request. Poster can be sent upon request in pdf format. Main sponsor of the Workshop is Nikon Italia, SpA, Firenze. News on the workshop can be found at www.lambs.it, from January 13th 2004. Further sponsorship can be proposed to diaspro-at-fisica.unige.it using as subject "confocal 5 - sponsor". All my best Alberto Diaspro ....................................................................... ................................... .. non basta, resistere, resistere...resistere ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Alberto Diaspro Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa, Italy voice: +39-0103536426/480 fax 010314218 diaspro-at-fisica.unige.it, http://www.lambs.it http://wiley.com/cda/product/0,,0471409200,00.html
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 06:43:27 2004
Electron Microscopist/Laboratory Manager Position Department of Materials Science and Engineering Drexel University, Philadelphia, PA 19010
We are seeking a candidate to supervise a microscopy laboratory, which includes an FEI/Phillips XL30 field emission environmental scanning electron microscope with an EDAX-EDS and TSL-OIM, an Amray 1830/D4, and a basic TEM (we outsource most TEM work), optical microscopes, sample preparation facilities, two X-ray diffractometers, etc. We are moving towards a centralized facilities model for the entire university and anticipate the move of many of our laboratories to a new research building by 2005. In addition to instrument maintenance, user scheduling, supervising and training users, the person in this position is expected to participate in the planning and execution of tasks related to the centralized materials testing and characterization facilities and the relocation of the labs to the new building. Candidates should have demonstrated experience with electron microscopy and preferably a degree in Materials, Physics or Biology. Salary will be commensurate with qualifications and experience.
The successful candidate will join a technical staff of three within the department. Detailed information about the department can be found at http://www.materials.drexel.edu, a copy of our recent newsletter can be found at http://www.materials.drexel.edu/Newsletter/fall_03.pdf and our last annual report at http://www.materials.drexel.edu/annualreports/Annual%20Report%202002.pdf. Candidates interested in this position should please submit a CV and a short career plan to: Microscopy Hiring Committee, Drexel University, Department of Materials Science and Engineering, 3141 Chestnut Street, Philadelphia, PA 19104.
Dr. Richard Knight -- "And the men who hold high places, must be the ones to start..."
Richard Knight, Ph.D., FASM, Auxiliary Professor, Center for the Plasma Processing of Materials [CPPM], Drexel University, Dept. of Materials Science & Engineering, 3141 Chestnut Streets, Philadelphia, PA 19104
A most interesting thread, and one that is near and dear to my heart.......
I frequently get people new to SEMs, either through personnel changes at a customer's site or through someone's acquisition of a "new" (used) SEM. Half of my business is still servicing ETEC Autoscans and as the new systems get more and more automated, I get more and more comments on how user unfriendly the Autoscan is. My standard reply is that it is not at all unfriendly if you understand how an SEM works.
I always start a new user out by building an SEM on paper and guess what? A scanning electron microscope is not a microscope at all! The only image formed in its optics is a DEmagnified image of the tip of the filament! So, what is it? It's a signal generator with one or more signal processors attached. What I really love about SEMs, especially with an EDS attached, is that you can tell me what you want to see and I can probably show it to you. Then we have to sit down and have a long talk about what is real. Don't tell me, "the computer said...". The computer doesn't know squat. And I haven't even gotten to specimen preparation!
The problem goes beyond the need for instant gratification, but may be related to the definition of success. People aren't interested in becoming microscopists because organizations no longer hire microscopists. One must pay too much for the knowledge and experience. Most are looking for "machine operators" that can be transferred between different pieces of equipment at will or they simply equate a microscope with a copier or personal computer. It's merely another tool to be used in getting the job done. We no longer have someone designated as the "copier specialist" in the office and having a degree in computer science is not a prerequisite for operating a computer on your desktop.
In part, there is simply so much more to know about any piece of equipment or area of interest, and the areas of interest and expertise are so intertwined, that one cannot be an expert in all areas, and in part it is the "Walmartization" of the world. The dive to the bottom, the least common denominator, the lowest cost, the lowest price. Of course, if everyone worked for Walmart, no one could afford to do much shopping............
I don't have any answers. I've always liked to understand any equipment that I use, whether it's a microscope or an automobile or a dishwasher, but then, I repair things for a living. I make a living this way because other people have different interests and drives (and that often does NOT include an interest or ability to repair things). A hundred years ago, most people who owned an automobile also had a mechanic in their employ, and the mechanic often traveled with, or even drove, the car.
I believe our problem in science will also get worked out. It's probably going to take some disaster related to bad science to get people's attention, but in the end, some kind of accommodation will be worked out. It will be interesting to watch it unfold.
In the mean time, we can all keep trying to spread our knowledge and our concerns. We subscribe to this listserver because we care. Keep caring!
Ken Converse owner Quality Images third party SEM service Delta, PA
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 07:15:40 2004
I would be careful with using a webcam for professional microscopy. Indeed, you can adjust a webcam this way that it can be attached to a microscope (remove the lens and put the ccd in the focuspoint of the lightbeam), but most webcams are very limited. Limited in shutterspeed, colorrange and fo sure in pixelresolution. Webcams are often used for amateur-microscopy and astronomy and give nice results, but if you really want to start analysing the (live) images or use them for publication, I'm afraid you might get dissapointed. Also: more expensive, professional camera's will last longer ,give you nicer resultes and a better support after sale will be offered. Best regards,
Sven Terclavers
On 08-01-2004 23:33, "Antonio Molina" {ifrm111-at-if.csic.es} 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 } ------------------------------------------------------------------------------} - } } Our old camera has to be replaced and I am offered a new one for 1200 euros. } This may not be too expensive but I wonder if I can get the same results } with a cheaper webcam attached to the microscope. I understand it is just a } question of removing the camera objective lense or to make it focus } infinity. } } My question is: can I expect a really better performance from a more } expensive, purpose-built camera? Quite likely the camera will not be the } limiting factor in the quality of micrographs, but other factors in the } microscope and the preparation... } } And, which factors should I cosider in the camera? I am thinking of } sensitivity to poor light, gain, and so. } } Thanks for all your comments, } } Antonio D. Molina-García } Inst. del Frio (CSIC) Madrid, Spain } } PD. My main purpose for video recording from a microscope is to study ice } crystal evolution during growth and recrystallization. Image is so, not too } sharp ever, as the contrast between ice crystals is small. Also the sample } thickness is larger than when using other sample types and, when the size } of } crystals is small, it is even difficult to get any light through... } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 07:31:01 2004
I very much agree with Sousan about what drives students and their reward systems. However, consider the fact that universities often reflect what is going on in private industry. I would bet that many researchers, scientists and engineers report to people that have scant knowledge of the science they ostensibly oversee. I call this the "MBA Phenomenon." I've observed this trend not only in microscopy. How often must someone in the lab. explain to their "management" exactly what an image is telling them? Quality should start with quality management and that means the management ought to know what they are managing.
Peter Tomic Agere Systems
Disclaimer: I am happy to report that the above conditions do not apply to me at the moment.
-----Original Message----- } From: Sousan Abolhassani [mailto:sousan.abolhassani-at-psi.ch] Sent: Thursday, January 08, 2004 4:29 AM To: John W. Raffensperger, Jr. Cc: microscopy-at-MSA.Microscopy.com
I fully agree with this fact that the problem is universal, and it is not only limited to microscopy. However, before one finds the "cure for this phenomenon", one should look for the cause of this problem.
If the new generation, i.e. the younger students and others, "are just wanting results", is it not the consequence of the ATTITUDE OF THE LARGE MAJORITY of those who trained them?
Those who trained the last two generations are now collecting the results of what they have done. The biggest reference for this generation is having rapid success, which means gaining large amounts of money and if possible a quick fame.
No university professor considers the students for their interest for science and their will to do something for the sake of humanity. They rank them on their good marks, without worrying about how they obtained it. And even if they do so, it is to REPRIMAND the student and not for TEACHING them the importance of honesty. A large percentage of students learn just to obtain a good mark, without thinking about why's and how's. And those few who do, are not appreciated by the professors, may be since they need more time that is not available.
A professorship is the art of teaching many things including highest human values to the students, I can not name many who have done that, or even have thought to do it. If they do not reflect such thoughts, at least they should avoid to discourage the very few who do have such values (and have truly come to learn something that can be helpful for the society) by treating them as naives and dealing with irony with them.
Some students tell me that the science professors are so arrogant that the students have no more appeal for scientific branches. } From my discussions with some undergraduate science students, their feeling is that if they go for science in their graduate school they could ruin their future and they will loose the chances of success!
I agree that the danger is real and from my understanding the solution to the problem is to reverse the trend from now on by changing the teaching philosophy. However, one should have the certitude of the importance of such a training to be able to have an impact on one's student.
Sousan Abolhassani, Ph.D. Laboratory for Materials Behaviour Paul Scherrer Institut 5232 Villigen PSI Switzerland
"John W. Raffensperger, Jr." 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 } ------------------------------------------------------------------------------- } } If it makes all of you feel any better, the phenomenon of users not } wanting to learn how to use something, just wanting the results, is not } limited to microscopy, and is a dangerous trend. This trend is } ironically aided by the very advancing technologies that make truly } understanding the theories and principles of what is happening more } important than ever. } } Too many devices, instruments and other systems have become "too easy to } use". The advances in human interfaces, automation, computer controls, } etc. have made it very easy for just about anyone to get results. The } danger is that there is no way for someone who doesn't truly understand } what's going on to know if the results are meaningful or not. "I got } the answer, and it's what I was expecting, so it must be right!" } } The problem is also not limited to "sophisticated" technology. We have } users who no longer know that there is a darkness adjustment on a copy } machine, much less how to use it. Not that long ago, you couldn't get } two copies in a row to turn out without tweaking the adjustment. If } someone does take the thing off "auto" and sets it dark, everyone else } thinks the thing is broken... } } If someone finds a "real" cure for this phenomenon, please share it with } the world, because the problem is pretty universal. } } John W. Raffensperger, Jr. } IS Manager } Helwig Carbon Products, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 08:13:05 2004
----- Original Message ----- } From: jamielange-at-wi.rr.com (by way of MicroscopyListServer)
The power of semantics
I am convinced that one of the reasons for a decline in the quality of microscopy, especially in industry, is semantic. Over the last few years it has become fashionable for managers to refer to instruments as "tools". They no longer distinguish between different kinds of equipment - they are all tools. Tools include: lathes, electron microscopes, fork lift trucks, x-ray diffraction units,.....
When a person using a tool leaves or is promoted, you need another person to operate the tool. So you take a person from this tool and put them on to that tool. Give them a couple of hours to read the manual and away you go.
I can only assume that machinists are complaining about the decline in the quality of work done in machine shops. All the ex-SEM operators must be doing a lousy job of operating the milling machines.
-- .......... 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 Jan 8 08:39:34 2004
In a message dated 1/8/04 9:28:40 AM, jae5-at-lehigh.edu writes:
} I can only assume that machinists are complaining about the decline in } } the quality of work done in machine shops. All the ex-SEM operators } must be doing a lousy job of operating the milling machines.
Alwyn, I don't think the analogy holds. Machine tools have gotten "smarter" in recent years. Numerical control and feedback devices have made it possible for information to flow directly from the design computers and CAD programs (which in turn have replaced the traditional draftsman) to control the machining process. In some cases the "operators" aren't even present, or just keep watch over a large array of machines in case of malfunctions or to load raw material. As the microscopes have evolved, they have for the most part not become easier to operate. Oh sure, some "little" things like adjusting astigmatism, saturating filaments, even focusing, have been automated. But not the "big" things like taking a meaningful picture. In fact, it has arguably become more complicated to use the modern microscopes because they offer a much broader range of possibilities than the old ones did. More imaging modes, more types of detectors, etc., create a greater demand for insight and knowledge on the part of the operator.
John Russ (visit www.DrJohnRuss.com for a schedule of upcoming image analysis workshops and other information)
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 08:50:32 2004
Mike Delannoy asked whether there is an easier way to make holey carbon films than Formvar, steam, create the holes in the damaged Formvar film, carbon coat the holey Formvar and dissolve the Formvar away to leave the holey carbon film. If there is, we'd like to know about it! This is the easiest method about which we know.
I must stress, however, that making holey and lacey carbon coated grids is pure art. People who have decades of experience making support films still have difficulty at times with the reproducibility of the method, and the details are entirely a matter of getting the "feel" of the method. Details of our methods are described on
http://www.2spi.com/catalog/grids/cusctgrd.html
and the pages linked to it.
An easier way to obtain holey carbon support films with a known and precise distribution of holes is the Quantifoil Micromachined Holey Carbon Grids, described on
Disclaimer: SPI Supplies has a very active business making coated grids for customers throughout the world. We also sell grids and other supplies for customers who prefer to coat their own grids, and we sell the Quantifoil Holey Carbon Grids.
Andy
Andrew W. Blackwood, Ph.D. Vice President, Technical SPI Supplies P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 X108 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:25:16 2004
While I see your point, it is much easier to at produce something with today's "tools", even your "tools". We can now use PhotoShop or some other application, load up some Image Tool filters and away you go with image analysis. My first IA package was very cumbersone and it took some major training just to load the images and navigate through the software. Presumably, you would pick some understanding of the subject matter through osmosis, solid state diffusion or entropy while learning how to operate the software. Now the software is so easy, anyone can sit down and after a few minutes start cranking out numbers.... for as meaningful or meaningless as they may be.
With SEMs, my first one filled a room and producing a photo was a real chore. You had to have an intimate knowledge of the controls and operating conditions to get even a poor quality photo. Now you can train a chipmunk (borrowed from one of my associates) to run an SEM. We have seen PhD's operate the SEM as a machine and have absolutely zero fundamental knowledge of the images or EDS data. I am thankful that I can sit down at my SEM and get a great photo in 5 minutes, but that means anyone else can too.
Al Stone ASTON
ps. no offense to anyone with a PhD, the point is that just because you can drive a car doesn't mean you understand the rules of the road or know how to travel cross country
At 08:42 AM 1/8/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:25:52 2004
My favorite quote fits well in this discussion: "It is not enough to believe what you see. You must also understand what you see. - Leonardo da Vinci"
I think it has to do a bit with what John just brought up in regards to the evolution of the microscopes. They have become "easier" to operate, and in doing so (computer control, mouse operated saturation...) there is a great disconnect with how the 'system' works. Users learning on an old EM300 for example have more interactions with the parameters on the microscope and thus are in a better position to 'understand' what they are doing in the process of collecting an image.
It is difficult to stress integrity, understanding when teaching. Even harder is the problem educators have in providing evaluation of the process, esp when there is a definitive end product (a lab report with images for example). I would much prefer to be able to grade students on their process than on the end results, and to provide an objective progress evaluation that translates onto a final grade. The biggest problem is that each student is different, has different learning speeds, different ways of coming to the same end point. So many variables to deal with. I find that the most effective and simple question I can present to students and faculty using the facility is "How do you know [ That ] is what you are looking at?" And sometimes it becomes a significant frustration level when the individual cannot convince me. But when they CAN convince me they wind up becoming much more confident and it forces them to find supporting evidence that they were too lazy or busy to look for earlier, and then they (more often than not) are much better at looking (literature sourcing) BEFORE starting their next project.
Well it is a new year and for some of us a new Semester. Time to start doing what ever we can do to reverse the trend!
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
} -----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] } Sent: Thursday, January 08, 2004 9:42 AM } To: jae5-at-lehigh.edu; microscopy-at-msa.microscopy.com } Subject: [Microscopy] Re: Re: Knowledge and Quality } } } } ------------------------------------------------------------------------ -- } ---- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ -- } ----- } } } In a message dated 1/8/04 9:28:40 AM, jae5-at-lehigh.edu writes: } } } I can only assume that machinists are complaining about the decline in } } } } the quality of work done in machine shops. All the ex-SEM operators } } must be doing a lousy job of operating the milling machines. } } Alwyn, I don't think the analogy holds. Machine tools have gotten } "smarter" } in recent years. Numerical control and feedback devices have made it } possible } for information to flow directly from the design computers and CAD } programs } (which in turn have replaced the traditional draftsman) to control the } machining } process. In some cases the "operators" aren't even present, or just keep } watch } over a large array of machines in case of malfunctions or to load raw } material. As the microscopes have evolved, they have for the most part not } become } easier to operate. Oh sure, some "little" things like adjusting } astigmatism, } saturating filaments, even focusing, have been automated. But not the } "big" things } like taking a meaningful picture. In fact, it has arguably become more } complicated to use the modern microscopes because they offer a much } broader range of } possibilities than the old ones did. More imaging modes, more types of } detectors, etc., create a greater demand for insight and knowledge on the } part of } the operator. } } John Russ } (visit www.DrJohnRuss.com for a schedule of upcoming image analysis } workshops } and other information)
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:28:16 2004
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Thursday, January 08, 2004 8:42 AM
In a message dated 1/8/04 9:28:40 AM, jae5-at-lehigh.edu writes:
} I can only assume that machinists are complaining about the decline in } } the quality of work done in machine shops. All the ex-SEM operators } must be doing a lousy job of operating the milling machines.
Alwyn, I don't think the analogy holds. Machine tools have gotten "smarter" in recent years.
-Reply:
I agree with Alwyn. This is the exact phenomenon I was referring to, and Machine Tools are one of the specific examples I had in mind with my original comments to this thread.
As you stated, there are shops where those involved truly understand what's going on, and things work very well.
In other shops, while these "tools" have indeed become "smarter", the quality of the output has decreased. This is because, as another post stated, these newer machines are viewed as a "tool", and an "operator" is assigned. Because of the intelligence of the machine, the "operator" who is no longer a "machinist" can get a result. It is often not an optimal result, either in terms of quality, and/or in terms of production rate. But a result was obtained. Compounding this, management has seen this situation "evolve" gradually, so they don't realize how much the "lower priced" help is actually costing them vs. a "real" machinist.
Add CAM instructions coming from an engineer who has never even operated a machine, and things get even more fun. Ask me about the $100,000+ damage one of these "wonder kids" did when they crashed a brand new machining center. The simulation ran perfectly. Too bad the engineer forgot that something had to hold the work piece, and this something was 2" think...
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:36:59 2004
We have a Zeiss 109 TEM available. Scope includes the unique SEM attachment (gives SEM capability with two SEM stages that fit standard Zeiss pin mounts). Turbo pumped; extra gun assembly, filaments, water chiller, manuals, and original orange paint on column included. Has been under full Zeiss/Leo service contract and in use until last month.
Needs to be removed as soon as possible to make room for a new scope.
Preference given to academic institutions. If you want it, you need to make arrangements for shipping, or pick it up in New London, CT. Contact me offline for further information.
Page Owen
************************ T. Page Owen, Jr., Chair Department of Botany Connecticut College New London, CT 06320 860-439-2147 tpowe-at-conncoll.edu ************************
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:46:30 2004
Without a picture of what you are seeing advice is difficult. My guess is that the section is not flat on the slide and water/stain is getting underneath a wrinkle in the section and the result is the "ribbon" you are seeing. Suggestions to solve the problem:
1. A sharp knife. If you are using glass knives use only knives made "fresh". Glass, being a super-cooled liquid, flows and older knives are less sharp. Try it! 2. De-wrinkle the sections with 1,2 dicholorethane or chloroform. 3. Float the sections on a larger drop of water on a hot plate. The larger drop will take longer to evaporate and give the section more time to be expanded/dewrinkled by heat. 4 Thinner sections? I don't know how thick your sections are but 1-2 microns is the range to shot for.
Geoff
by way of MicroscopyListServer wrote:
} Email: jamielange-at-wi.rr.com } Name: jamie lange } } Organization: university of wisconsin } } Education: Graduate College } } Location: milwaukee, wi. usa } } Question: we used a spurr's resin prep on a sample of nematodes, after } staining and heat-fixing the sections, we see dark ribbon-like } structures on several specimens. Are these artifacts caused by air } bubbles and can they be avoided? thank you, } jamie } } --------------------------------------------------------------------------- } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:52:27 2004
First of all EVERYONE must realise that the image that they see (even though they pressed the SE or SEI button) is a mixed SE+BSE image. This means surface detail from the SE and sub surface detail from the BSE. As you put the kV up the BSE dominates more and of course as you take the kV down the BSE are reduced allowing the SE to dominate. I should point out that you will often see BSE information even at 2kV or less; this being the result of the SE3 contribution.
Place a thin coating on the surface of a specimen and you increase the coefficient of emission, the metal being the SE emitter rather than the original biological material. You do image the specimen surface as that topography has been followed by the coating procedure. However the BSE come from a far greater depth below the surface and at any kV, under the wrong circumstances of WD and spot size, these electrons will contribute to the image. Should there be structure of differing density, by way of the BSE, this will show as "shadows" in the background or may even dominate the image.
In a field emission instrument, of the type where the above lens detector is available, it is possible to screen out the BSE contribution and display a pretty pure SE image. I have to say in my 40 years in the business we have more often gone for "information" rather than "resolution" therefore I do try to include a degree of BSE to add contrast to the image.
Hope this helps?
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "Tobias Baskin" {baskin-at-bio.umass.edu} To: "Steve Chapman" {protrain-at-emcourses.com} ; {microscopy-at-MSA.microscopy.com} Sent: Wednesday, January 07, 2004 4:51 PM
It appears we have a change in how quality is monitored over the years of SEM evolution. As may be the case where older SEMs require understanding of how a SEM works to get an image, to the extent that newer SEMs do not require this, those operating newer SEMs may not require the prior extent of knowledge to produce images. Where once the decision to take images were made by those who knew enough to understand and self monitor quality, that appears not to be the case anymore. Advances in technology have allowed images to be taken by operators who do not understand what they are looking at; one can not expect an unknowing individual to monitor quality.
We can address the quality issue with education or attitude (as proposed by others on this board). This is a head on approach; a less direct method would be to change the decision rights or incentives. For example, operators should not be given the decision right of what to take an image of. For example, operators should not be rewarded simply on the number of images taken.
The point is that monitoring of quality has changed, so we should consider changing decision rights and incentives/rewards. If these three components of organizational architecture are not in balance, results will be unsatisfactory.
Sincerely, John Moore Montara Industries 919-434-8457
Disclaimers: 1) This framework is described in a book titled "Managerial Economics and Organizational Architecture", a text that I studied while obtaining my MBA at the University of Rochester, Class of 2000.
2) In defense of my education and this technique, I cite first that the poster from Agere on this topic reported not suffering certain difficulties, and second that in my recent job search I found Agere to be hiring MBA's (see monster.com directemployers.com, hotjobs.com or flipdog.com).
3) I am not a SEM expert, nor do I have direct experience in this field. I began following this message board in the interest of selling a defect review tool that I made a speculative investment in: a SEMVision by Applied Materials.
----- Original Message ----- } From: "Sousan Abolhassani" {sousan.abolhassani-at-psi.ch} To: "John W. Raffensperger, Jr." {johnr-at-helwigcp.com} Cc: {microscopy-at-MSA.Microscopy.com} Sent: Thursday, January 08, 2004 4:28 AM
I fully agree with this fact that the problem is universal, and it is not only limited to microscopy. However, before one finds the "cure for this phenomenon", one should look for the cause of this problem.
If the new generation, i.e. the younger students and others, "are just wanting results", is it not the consequence of the ATTITUDE OF THE LARGE MAJORITY of those who trained them?
Those who trained the last two generations are now collecting the results of what they have done. The biggest reference for this generation is having rapid success, which means gaining large amounts of money and if possible a quick fame.
No university professor considers the students for their interest for science and their will to do something for the sake of humanity. They rank them on their good marks, without worrying about how they obtained it. And even if they do so, it is to REPRIMAND the student and not for TEACHING them the importance of honesty. A large percentage of students learn just to obtain a good mark, without thinking about why's and how's. And those few who do, are not appreciated by the professors, may be since they need more time that is not available.
A professorship is the art of teaching many things including highest human values to the students, I can not name many who have done that, or even have thought to do it. If they do not reflect such thoughts, at least they should avoid to discourage the very few who do have such values (and have truly come to learn something that can be helpful for the society) by treating them as naives and dealing with irony with them.
Some students tell me that the science professors are so arrogant that the students have no more appeal for scientific branches. } From my discussions with some undergraduate science students, their feeling is that if they go for science in their graduate school they could ruin their future and they will loose the chances of success!
I agree that the danger is real and from my understanding the solution to the problem is to reverse the trend from now on by changing the teaching philosophy. However, one should have the certitude of the importance of such a training to be able to have an impact on one's student.
Sousan Abolhassani, Ph.D. Laboratory for Materials Behaviour Paul Scherrer Institut 5232 Villigen PSI Switzerland
"John W. Raffensperger, Jr." 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 } -------------------------------------------------------------------------- ----- } } If it makes all of you feel any better, the phenomenon of users not } wanting to learn how to use something, just wanting the results, is not } limited to microscopy, and is a dangerous trend. This trend is } ironically aided by the very advancing technologies that make truly } understanding the theories and principles of what is happening more } important than ever. } } Too many devices, instruments and other systems have become "too easy to } use". The advances in human interfaces, automation, computer controls, } etc. have made it very easy for just about anyone to get results. The } danger is that there is no way for someone who doesn't truly understand } what's going on to know if the results are meaningful or not. "I got } the answer, and it's what I was expecting, so it must be right!" } } The problem is also not limited to "sophisticated" technology. We have } users who no longer know that there is a darkness adjustment on a copy } machine, much less how to use it. Not that long ago, you couldn't get } two copies in a row to turn out without tweaking the adjustment. If } someone does take the thing off "auto" and sets it dark, everyone else } thinks the thing is broken... } } If someone finds a "real" cure for this phenomenon, please share it with } the world, because the problem is pretty universal. } } John W. Raffensperger, Jr. } IS Manager } Helwig Carbon Products, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 10:39:09 2004
To Joel, Eckhart, Kenneth, Peter, and anyone else who gave me advice-
OK, I finally had a chance to sit down and fiddle with the chiller, and here is what I have.
I turned on my Zeiss EM 10CA and checked the flowmeter in the column: about 2.2 L/min, if I am reading it right. There was no gauge for the chiller flow rate, but there is a pressure gauge on the front of the chiller, and it read about 39 psi. This reading did not change through the entire test, and neither did the EM flowmeter.
Chiller temperature reading was about 75 degrees F at start. It needs to be 68 degrees F, according to the EM's manual. Only the "Cool" light was on...all others, including the "Compressor" light, were off.
I turned the temperature screw to the left to lower the temp setting. No response from the compressor. Turned it down a second time....still no response. After about 5 minutes the Compressor light turned on, and the temperature started to drop, finally. The temperature went all the way down to 53 degrees F, at which point the "Lo Temp" light went on, the "Compressor" light went off, and the "Heat" light went on. I turned up the temp screw a little bit.
At about 60 degrees F, the "Heat" and "Lo Temp" lights went off and the "Cool" and "Pump" lights are on (actually the "Pump" light never turned off). It rose to 68 degrees F....and continued to rise. I let it get to about 79 degrees F, and the compressor never turned back on. I tried turning the screw back to the left to drop the temp setting...it never went back on.
At this point, I decided to turn the EM 'scope off....though the flow rate never dropped, and the buzzer never went off. The temperature gauge on the chiller continued to rise, even after turning the 'scope off. It was reading about 90 degrees F when I left to come write all of you.
Just now went back in to check the thing...the compressor finally came on...it's reading about 53 degrees F and the "Lo Temp" light is on, and the "Compressor" light is off.
So I think it's the switch....or perhaps the "Hi Temp" sensor? Or both? I have no idea how all the sensors and switches are connected...even with the diagrams and manual that were sent, as I have no idea how to read such things (been eons since I did circuits in Physics class...). What do you all think?
Thank you all so much for your help- Kathleen Neurotoxicology Labs Rutgers University
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 12:23:55 2004
I've been reading the iterations on this thread and it seems to me that it would be useful to separate the chiller from the TEM to test the chiller. Kathleen, if you have a carbon evaporator that has a diffusion pump you can cool it with the Coolwell long enough to tell if the Coolwell has a problem. If the Coolwell tests okay on another heat load (the carbon evaporator) then your problem is in the TEM.
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
At 11:48 AM 1/8/2004 -0500, Kathleen Roberts wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 13:00:36 2004
Well some interesting stuff, I was particularly taken by Peter Tomic's comments
"Quality should start with quality management and that means the management ought to know what they are managing."
Via MSA and mostly privately I have had mail from, judging by the mailers enthusiasm, pretty good EM units. However I have probably visited more EM units in the world than most of you and I have to say in very many cases they are just not up to the standards that I would set. Here Peter's comment hits home to me! You see, as an example, in most units people were trained when the SEM, TEM or EDS system first arrived in the lab. Now they have new equipment and I bet they have rarely been trained to operate using the techniques opened up by the new SEM or TEM? Once trained you know how to use them is what I see. Guess what, a customer born on a Cambridge 100 SEM still used their new sexy Field Emission SEM in just the same way. A thick gold coating and 20kV plus was the way to do it, quote "we have always done it this way". We produced amazing results uncoated at 150,000X at 1.5kV! Do I make my point? This was a very highly rated government research laboratory that by perception would be rated as in the top 5 in a country very well respected for its EM work!
We all believe we run a good unit, my point would be "how do you know?" Do you have management procedures that are up to date, do you routinely check the performance of your staff, do you look at the output your users produce, etc etc? Take a look at www.iaaem.com/mppa1 here you will find a quality procedure that has been proposed by a group of scientists and Quality experts from across the world. Answer the questions and total up the points for your laboratory and lets see how far you area away from Total Quality Management. What would you suggest is included and maybe together we will be able to develop procedures that would help sort out what you all, almost, seem to agree is a bit of a mess!
I am working overseas (Egypt) for the next week so have fun on your own for a little while.
Regards
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 13:44:05 2004
I want to give away the following to a university or non-profit lab or group:
Plexiglass racks for holding 3.25" x 4" (TEM) and 4" x 5" (Polaroid) negative film; and hard rubber tanks for processing negatives.
Interested parties should contact me off-line. Commercial/industrial users need not respond.
Thanks,
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 14:04:04 2004
Good idea, except that I don't have a carbon evaporator. :o)
Thanks anyway, Kathleen Neurotox Labs Rutgers University
Owen P. Mills wrote:
} I've been reading the iterations on this thread and it seems to me } that it would be useful to separate the chiller from the TEM to test } the chiller. Kathleen, if you have a carbon evaporator that has a } diffusion pump you can cool it with the Coolwell long enough to tell } if the Coolwell has a problem. If the Coolwell tests okay on another } heat load (the carbon evaporator) then your problem is in the TEM. } } Owen } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills } } At 11:48 AM 1/8/2004 -0500, Kathleen Roberts 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 } } ------------------------------------------------------------------------------- } } } } } } To Joel, Eckhart, Kenneth, Peter, and anyone else who gave me advice- } } } } OK, I finally had a chance to sit down and fiddle with the chiller, } } and here is what I have. } } } } I turned on my Zeiss EM 10CA and checked the flowmeter in the column: } } about 2.2 L/min, if I am reading it right. There was no gauge for } } the chiller flow rate, but there is a pressure gauge on the front of } } the chiller, and it read about 39 psi. This reading did not change } } through the entire test, and neither did the EM flowmeter. } } Chiller temperature reading was about 75 degrees F at start. It } } needs to be 68 degrees F, according to the EM's manual. Only the } } "Cool" light was on...all others, including the "Compressor" light, } } were off. } } } } I turned the temperature screw to the left to lower the temp setting. } } No response from the compressor. Turned it down a second } } time....still no response. After about 5 minutes the Compressor } } light turned on, and the temperature started to drop, finally. The } } temperature went all the way down to 53 degrees F, at which point the } } "Lo Temp" light went on, the "Compressor" light went off, and the } } "Heat" light went on. I turned up the temp screw a little bit. } } } } At about 60 degrees F, the "Heat" and "Lo Temp" lights went off and } } the "Cool" and "Pump" lights are on (actually the "Pump" light never } } turned off). It rose to 68 degrees F....and continued to rise. I } } let it get to about 79 degrees F, and the compressor never turned } } back on. I tried turning the screw back to the left to drop the temp } } setting...it never went back on. } } } } At this point, I decided to turn the EM 'scope off....though the flow } } rate never dropped, and the buzzer never went off. The temperature } } gauge on the chiller continued to rise, even after turning the 'scope } } off. It was reading about 90 degrees F when I left to come write all } } of you. } } } } Just now went back in to check the thing...the compressor finally } } came on...it's reading about 53 degrees F and the "Lo Temp" light is } } on, and the "Compressor" light is off. } } So I think it's the switch....or perhaps the "Hi Temp" sensor? Or } } both? I have no idea how all the sensors and switches are } } connected...even with the diagrams and manual that were sent, as I } } have no idea how to read such things (been eons since I did circuits } } in Physics class...). What do you all think? } } } } Thank you all so much for your help- } } Kathleen } } Neurotoxicology Labs } } Rutgers University } } } } } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills }
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 14:12:46 2004
In my experience, CCTV cameras as used in security applications work better than webcams, but the resolution of both is not that great and you may spend considerable time on the mechanical aspects of the interfacing and be disappointed with the result.
1200 euros may be well worth it for something that you can just unpack from the box and be using in a few minutes.
cheers
rtch
Send reply to: "Antonio Molina" {ifrm111-at-if.csic.es} } From: "Antonio Molina" {ifrm111-at-if.csic.es} To: {Microscopy-at-MSA.Microscopy.Com}
Most people store UA in the refrigerator perhaps without understanding why. UA is photosensitive and degraded by light (especially fluorescent lighting that contains UV). If you store aqueous solutions in the light they will eventually precipitate --first long the sides of the container and then on the bottom. I have no data, just based on observation.
} We are having one of those debates that we microscopists seem to } obsess about. The question is whether to store our saturated uranyl } acetate solution (in dH2O) at room temp or at 4 C. Opinions, } especially those backed by data, would be welcome. Happy New Year. } Tom } } } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
-- ############################################################## 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 Jan 8 14:31:49 2004
Thank you for the honorable mention. If my boss is on this listserver I'll have to polish up the old resume. It's so much fun to be quoted. I feel like Hemingway.
Once again, I don't have ANY of these issues HERE.
I needed a good laugh today.
Peter Tomic Unknown Corporation, Inc. Anytown, USA
-----Original Message----- } From: Steve Chapman [mailto:protrain-at-emcourses.com] Sent: Thursday, January 08, 2004 1:59 PM To: MSA
Hi Listers
Well some interesting stuff, I was particularly taken by Peter Tomic's comments
"Quality should start with quality management and that means the management ought to know what they are managing."
Via MSA and mostly privately I have had mail from, judging by the mailers enthusiasm, pretty good EM units. However I have probably visited more EM units in the world than most of you and I have to say in very many cases they are just not up to the standards that I would set. Here Peter's comment hits home to me! You see, as an example, in most units people were trained when the SEM, TEM or EDS system first arrived in the lab. Now they have new equipment and I bet they have rarely been trained to operate using the techniques opened up by the new SEM or TEM? Once trained you know how to use them is what I see. Guess what, a customer born on a Cambridge 100 SEM still used their new sexy Field Emission SEM in just the same way. A thick gold coating and 20kV plus was the way to do it, quote "we have always done it this way". We produced amazing results uncoated at 150,000X at 1.5kV! Do I make my point? This was a very highly rated government research laboratory that by perception would be rated as in the top 5 in a country very well respected for its EM work!
We all believe we run a good unit, my point would be "how do you know?" Do you have management procedures that are up to date, do you routinely check the performance of your staff, do you look at the output your users produce, etc etc? Take a look at www.iaaem.com/mppa1 here you will find a quality procedure that has been proposed by a group of scientists and Quality experts from across the world. Answer the questions and total up the points for your laboratory and lets see how far you area away from Total Quality Management. What would you suggest is included and maybe together we will be able to develop procedures that would help sort out what you all, almost, seem to agree is a bit of a mess!
I am working overseas (Egypt) for the next week so have fun on your own for a little while.
Regards
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 14:48:50 2004
Yes, alas, we have also seen the "take the picture and run" mentality here as well.
Like most microscopists who came up through the system, most of us eagerly learned everything related to microscopy (specimen prep, knife making, ultramicrotomy, alignment, optimization of the scope, darkroom, image interpretation, etc). It was fun and we enjoyed learning and making discoveries. Now, it seems the thrill is gone and the object is to get into the job market and make big bucks....
MONEY is the motivator and it can be used to influence people. For example, I point out that microscopy is a marketable skill and I prove it by giving the names of our former students (in biological and physical sciences) who got jobs in their discipline since they could do microscopy. An employer will generally hire the individual with the better set of skills. Unless they are incredibly naive or just plain stupid (in which case you wouldn't want them using your equipment anyway) they will realize the value in learning the discipline.
More immediately, I point out that it is CHEAPER for them to do their own microscopy than to hire it out.
-- ############################################################## 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 Web: http://www.siu.edu/~image/ ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 15:10:24 2004
I was wondering if someone knows where I can find an monochromator for spectral scanning 400-700 nm that can be attached to a Zeiss microscope (Axioskop) for transmitted light imaging ( scanning microphotometry).
On Jan 8, 2004, at 6:42 AM, DrJohnRuss-at-aol.com wrote:
} Alwyn, I don't think the analogy holds. Machine tools have gotten } "smarter" } in recent years. Numerical control and feedback devices have made it } possible } for information to flow directly from the design computers and CAD } programs } (which in turn have replaced the traditional draftsman) to control the } machining } process. In some cases the "operators" aren't even present, or just } keep watch } over a large array of machines in case of malfunctions or to load raw } material. Dear John, So now the machine tools are capable of performing as well as the automated tomography packages we use? In our case, we have to watch the progress of the program carefully to see that the auto-tracking, auto-focussing, etc. is working properly, and lately we have also found that the file made from the tilt series has values that do not match the exposure we set (to the extent that some of the images were blank). The take-home lesson is that there still must be knowledgeable oversight, especially with regard to automated processes, to assure the quality of the results. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 15:14:33 2004
A lab user has asked me to find some place he can get freeze fracture work done. Close to San Francisco bay area would be best. Reply to me and I will pass on the contact info.
Thanks
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 Thu Jan 8 15:34:44 2004
Dear Kathleen, I have had good luck getting a local HVAC (Heating, Ventilation and Air conditioning) or Air Conditioning repair shop to fix my Haskris water chillers. Haskris are good but they do occasionally break down. Sounds like one of your sensors or accuators is stuck. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Kathleen Roberts" {kgrobert-at-rci.rutgers.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 08, 2004 8:48 AM
Thank you for the advice...I figure $10 isn't too much to spend to try and fix this thing....if the repair cost starts to escalate, we'll junk it and call Haskris. :o)
Kathleen Neurotoxicology Labs Rutgers University
Webster, Paul wrote:
} Hi Kathleen, } } A word of warning if you plan to try to fix the chiller. We spent nearly $7,000 in a year of repairing our Coolwell chiller before it finally failed for good and we had to buy a new machine. When the second one started giving problems, we used the repair money to buy a new one immediately. } } The first one started by developing a leak in the compressor, then the pump failed and finally the thermostat switch. Getting a similar thermostat switch to the one that failed was impossible. We searched through Grainger, talked with refrigeration engineers and could only come up with a close approximation of what we needed. } } We had to suffer as the less sensitive thermostat switch attempted to hold the water temperature steady but with much less control. In the end it was when some other part of the machine failed that we decided to replace it. Looking back, the time and expense needed to repair the machine was not worth it. } } If you do find your chiller has only a small problem then try fixing it and hope the costs don't escalate. } } Regards, } } Paul Webster. } } } } } Paul Webster, Ph.D. } House Ear Institute } 2100 West Third Street } Los Angeles } CA 90057 } phone (213) 273 8026 } fax (213) 413 6739 } email: pwebster-at-hei.org } } } } } } ---------- } } From: Kathleen Roberts } } Sent: Thursday, January 8, 2004 12:13 PM } } To: Owen P. Mills } } Cc: Microscopy-at-sparc5.microscopy.com } } Subject: [Microscopy] Re: Coolwell chiller testing } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 16:00:34 2004
Nope, no SEM and no BIG waterbath. I'm going to call Facilities here at RU-they may be able to devise something to test it.
Thanks- Kathleen Neurotoxicology Labs Rutgers University
Owen P. Mills wrote:
} Kathleen, } } Any heat source that can be water cooled will work, anything with a a } diffusion pump. SEM? Got a BIG hot water bath? Coil copper tubing } in the bath, turn the bath heat way up, connect the chiller lines to } the ends of the coil of copper tubing. Should be a low tech test of } the chiller. } } Owen } } At 03:13 PM 1/8/2004 -0500, you wrote: } } } Owen- } } } } Good idea, except that I don't have a carbon evaporator. :o) } } Thanks anyway, } } Kathleen } } Neurotox Labs } } Rutgers University } } } } Owen P. Mills wrote: } } } } } I've been reading the iterations on this thread and it seems to me } } } that it would be useful to separate the chiller from the TEM to test } } } the chiller. Kathleen, if you have a carbon evaporator that has a } } } diffusion pump you can cool it with the Coolwell long enough to tell } } } if the Coolwell has a problem. If the Coolwell tests okay on } } } another heat load (the carbon evaporator) then your problem is in } } } the TEM. } } } } } } Owen } } } } } } Owen P. Mills } } } Electron Optics Engineer } } } Materials Science & Engineering } } } Michigan Technological University } } } Rm 512 M&M Bldg. } } } Houghton, MI 49931 } } } PH 906-369-1875 } } } FAX 906-487-2934 } } } mailto:opmills-at-mtu.edu } } } http://www.mm.mtu.edu/~opmills } } } } } } At 11:48 AM 1/8/2004 -0500, Kathleen Roberts 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 } } } } ------------------------------------------------------------------------------- } } } } } } } } } } } } To Joel, Eckhart, Kenneth, Peter, and anyone else who gave me advice- } } } } } } } } OK, I finally had a chance to sit down and fiddle with the chiller, } } } } and here is what I have. } } } } } } } } I turned on my Zeiss EM 10CA and checked the flowmeter in the } } } } column: about 2.2 L/min, if I am reading it right. There was no } } } } gauge for the chiller flow rate, but there is a pressure gauge on } } } } the front of the chiller, and it read about 39 psi. This reading } } } } did not change through the entire test, and neither did the EM } } } } flowmeter. } } } } Chiller temperature reading was about 75 degrees F at start. It } } } } needs to be 68 degrees F, according to the EM's manual. Only the } } } } "Cool" light was on...all others, including the "Compressor" light, } } } } were off. } } } } } } } } I turned the temperature screw to the left to lower the temp setting. } } } } No response from the compressor. Turned it down a second } } } } time....still no response. After about 5 minutes the Compressor } } } } light turned on, and the temperature started to drop, finally. The } } } } temperature went all the way down to 53 degrees F, at which point } } } } the "Lo Temp" light went on, the "Compressor" light went off, and } } } } the "Heat" light went on. I turned up the temp screw a little bit. } } } } } } } } At about 60 degrees F, the "Heat" and "Lo Temp" lights went off and } } } } the "Cool" and "Pump" lights are on (actually the "Pump" light } } } } never turned off). It rose to 68 degrees F....and continued to } } } } rise. I let it get to about 79 degrees F, and the compressor never } } } } turned back on. I tried turning the screw back to the left to drop } } } } the temp setting...it never went back on. } } } } } } } } At this point, I decided to turn the EM 'scope off....though the } } } } flow rate never dropped, and the buzzer never went off. The } } } } temperature gauge on the chiller continued to rise, even after } } } } turning the 'scope off. It was reading about 90 degrees F when I } } } } left to come write all of you. } } } } } } } } Just now went back in to check the thing...the compressor finally } } } } came on...it's reading about 53 degrees F and the "Lo Temp" light } } } } is on, and the "Compressor" light is off. } } } } So I think it's the switch....or perhaps the "Hi Temp" sensor? Or } } } } both? I have no idea how all the sensors and switches are } } } } connected...even with the diagrams and manual that were sent, as I } } } } have no idea how to read such things (been eons since I did } } } } circuits in Physics class...). What do you all think? } } } } } } } } Thank you all so much for your help- } } } } Kathleen } } } } Neurotoxicology Labs } } } } Rutgers University } } } } } } } } } } Owen P. Mills } } } Electron Optics Engineer } } } Materials Science & Engineering } } } Michigan Technological University } } } Rm 512 M&M Bldg. } } } Houghton, MI 49931 } } } PH 906-369-1875 } } } FAX 906-487-2934 } } } mailto:opmills-at-mtu.edu } } } http://www.mm.mtu.edu/~opmills } } } } } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills }
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 16:02:07 2004
In a message dated 1/8/04 4:44:05 PM, tivol-at-caltech.edu writes:
} The take-home lesson is that there still must be knowledgeable } oversight, especially with regard to automated processes, to assure the } quality of the results
You'll get no argument from me about that! The point I was trying to make is that as the tools have become more complex, many of the tasks that we once dealt with manually (I learned electron microscopy on a Siemens 1A, right on the Caltech campus, back in the '50's - and as a result I really know what alignment means, not just how to push a button) are now so automated that they are hard to control. We may (but may not) be able to spot problems, but the casual user (shudder) will not know how to correct them.
I think this thread has been interesting primarily in that everyone who has commented has been in basic agreement that far too many people who use microscopes understand them, or the ancillary techniques of specimen preparation, image analysis, etc., well enough to keep out of trouble or get really optimum results. Clearly they have other priorities than learning all that stuff. I've taught image analysis courses now to something approaching 3500 people. Even assuming they all learned everything I wanted them to, that is a drop in the bucket. And the people who really need it most don't come - at best they send a technician whom they can subsequently tell to do the work. But that's another rant.
John Russ
John Russ
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 17:36:30 2004
} A lab user has asked me to find some place he can get freeze fracture } work } done. Close to San Francisco bay area would be best. Reply to me and I } will } pass on the contact info. } Dear Jon, If Kent McDonald does not have freeze-fracture, he probably knows someone in or near Berkeley who does. I don't have his email address immediately at hand. 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 Fri Jan 9 03:06:04 2004
The same is with EPMA (Electron Probe Microanalysis) with EDS. Unfortunately m o s t manufacturers moved their goals and tried to develop calculation machines as 'black box'. Because of the giant improvements in calculation speed of our computers (in last decades), only one button hit is necessary to display the complete analysis result (in most cases computer needs a tiny part of a second). The unskilled or less skilled operators believe in these results, without any concern. Even element concentration result errors of 0.1% and less are taken from the computer display as the truth. But such a very high 'accuracy' must be only the statistical part! The computer speed and modern easy to use software interfaces cover the very complex and not linear relations between measured signal and element concentration in specimen. The iteration process to get result convergence and the systematic and statistic errors with their error propagation during computing process are not visible.
I think for future, a more open software is going to be a trend. There must be a possibility to interact between the knowledge of the microanalyst and the computer program. A visible and easy understandable feedback for all computing steps is necessary. Of course, a higher skilled level of the operator is then necessary. This makes sense only, if the software give the possibility to share the knowledge with the operator, which is then really become a microanalyst.
A couple of years ago, I found in a very old German book of C. Remigius Fresenius "Anleitung zur Qualitativen Chemischen Analyse" ("Guidance to the Qualitative Chemical Analysis") - Braunschweig 1874:
"Es muss daher ein Halbwissen, wie überall so ganz besonders hier, für schlimmer als ein Nichtwissen erachtet und vor oberflächlicher Beschäftigung mit der chemischen Analyse ganz vorzüglich gewarnt werden."
Translation (I hope the feeling is transfered): Therefore a partial knowledge, like everywhere so particularly here, must be worse than even ignorance. It should be warned before superficial concern with the chemical analysis completely and excellently here.
These words are still valid and can be used nowadays for EPMA and Electron Microscopy including image interpretation, as well.
Frank Eggert
Steve Chapman wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Jan 9 05:35:11 2004
My two cents worth on a subject that has quite a few here tossing their own around.
Having spent 25 years in this area, I've had the opportunity to see the initial investigations of this practical application of sub-atomic particle physics in basic research labs as well as its resultant spread to a wide variety of industries. The use of SEM analysis has become a commodity because it is so successful in so many areas. Decades ago, many were researching the potential applications, and in virtually each case, they found the usefulness.
I can't begin to tell you the variety of areas where my customers have found the SEM useful. From missle guidance systems to electron beam lithography. Particulate analysis of howitzer oils to particulate contaminant analysis of sausage casings. The taxonomical classification of emerging bacterial species to process control for the laser produced hologram labels used for software copyright and federal IDs.
The fact is that the SEM has perhaps been too successful. We still hold in reverence the atomic physicists who can design an atomic bomb or understand the results from particle accelerators. But the SEM is a child of these processes that has found wide spread use and the demands for its application greatly exceed the number of those who really understand the underlying processes, much less the proper application of the various subtleties of its application.
I've had to deal with this over the years as the operators I deal with have changed from those individuals who first brought an SEM into a lab to the 'checklist' operators who know nothing about the instruments other than the written sequence of actions to turn it on and get an image. One of the challenges in my work is to try to encourage the SEM operators to learn and think more about the consequences of each turn of a knob or click of a mouse. As a maintenance provider, it certainly helps me if my customers have some understanding of their machines - but more importantly, it helps them do better work. SEMs aren't the only problems here - x-ray spectroscopy in fluorescence or the SEM based microanalysis is another area where operators are often fooled into thinking that results are a simple matter of a set routine.
The computerization of the instruments is furthering the problem. While the manufacturers are seeking only to play to the market, how many operators really understand what's happening when they tell their computer to bring up an FE gun? It really started with simple improvements such as electronic gun adjustments. When an operator had to physically move the gun assembly around, it made some sense that the position of the gun was important to aim the beam down the column. How many really understand the use of magnetic fields in the gun to tilt and shift the beam to alignment? Most that I see at first only know that they have to tweak these knobs and watch for an improvement in the signal.
Like it or not, this trend will continue. But it is not selective to SEMs - I see similar trends in every analytical instrument. As these instruments become more 'user friendly', they are actually lulling users into thinking that all that is needed is a brief glimpse at the user manual, which usually only describes how to push the buttons. IR, GC, LC, MS, ICP and many other techniques that involve complex physics have been reduced to a simplicity that masks their proper use primarily because those using them and buying them want simple answers. A material scientist investigating ceramics doesn't want to have to learn the sub-atomic particle interactions involved, he just wants pretty pictures that explain a manufacturing fault and justify the expense of the SEM, not to mention his job.
'Ease of use' is a marketing tool, and as such, it is a primary goal of manufacturers. I don't mean to focus on them, because it is a vicious circle - the customers are demanding it, the manufacturers simply provide it. In this process, though, what gets lost is that the proper use and interpretation of these instruments requires more than the customers are wanting to afford and more than the manufacturers are wanting to admit to.
Now Steve's attention is a little more esoteric - the quality control of a reviewed paper. But doesn't that just follow from the above? The results, rather than the process, are what matter most now. More and more we see examples in studies that are published, only to be later refuted. NASA's claim a couple of years ago about evidence of ancient bacteria in Mars based rock found in the Antarctic has, last I knew, been lost in dispute. Pons and Fleischman, and Gallo, are of course extremes, but how much of what is accepted as reputable science has later fallen as poor science. A brief look at medical headlines over the past decade or two can give a good glimpse. Science itself is supposed to ensure honest and accurate results, and the assumption of most people, scientists included, is that anything purporting to be science, promoted by supposed scientists, has some truth. Innocent until proven guilty, so to say.
Whether authored by lack of knowledge of instrumental techniques, lack of personal integrity or poor selection of measured variables, many papers get published that should have been caught by reviewers. That's assuming that those reviewers are well versed in all aspects of a particular paper. But given the wide variety of instrumental techniques available today, it can be a daunting task to find a single person expert in all of the instrumental techniques presented in a paper, not to mention the basic field of the paper and mathmatical aspects. If a reviewer isn't well versed in all aspects and techniques of a particular paper, can he be expected to catch the kind of cross-discipline problem in your example? Since much goes unsaid in virtually any paper, should reviewers be required to request all details of sample provenance - the collection, preparation and analysis?
By the way, Steve, was there any mention, in the example you cited, whether the sample was coated or not, or is it an assumption of yours that is wasn't?
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 ph. (630) 513-7093 fax (630) 513-7092 web http://www.sem.com
-----Original Message----- } From: Steve Chapman [SMTP:protrain-at-emcourses.com] Sent: Wednesday, January 07, 2004 6:42 AM To: MSA
Hi Listers
As many of you may know I run a training organisation that travels world wide spreading the word on SEM, TEM and EDS operation. I also have a deep interest in "Quality in Electron Microscopy"as those who picked up our paper last year would know?
Now to the point. Once again I am picking up respected journals and finding examples of what I would call poor microscopy, but in truth it also demonstrates poor quality control!
To bring one image to mind. The micrograph is of a structure which is described as being an example of a smooth surface on a biological material. But, the micrograph was taken at 20kV, where the vast majority of the information will have come from beneath the surface softening the true surface detail.
First to remove the training aspect . Operators of SEM should be taught that by manipulation of kV and working distance one may subdue or enhance surface features. To use more than 10kV on most biological samples is asking for sub surface detail, ignore this and comments on surface irregularities are null and void in my mind. (I have to say I would probably try to use 2 to 5kV if the microscope used was produced in the last 15 years!)
Now the quality aspect. By the time a paper is published a number of steps should have been taken. Working backwards, the publisher should have the paper vetted by knowledgeable scientists who would be able to pick out the problems that I see and have them corrected prior to going to print. Next back in the chain is the laboratory that was involved with the scientist; did they check the quality of the work leaving their EM unit? Stepping back again did the scientist take the micrographs or did they receive help? Either way the training of staff and operators should overcome this type of problem! But if the results the staff and visiting operators produce are not assessed how do you know that their training is inadequate?
As the pressure to perform increases and funding decreases only the cream of our laboratories will remain. In industry there is no question about following rigid "Quality" procedures and it is not too far off that this will hit the world's EM units too! I know that this is my baby but is it not about time that we woke up to these facts?
There is an area where I believe we have room for discussion; what do you think?
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob +44 (0) 7711 606967 www.emcourses.com
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 9 08:38:55 2004
I cannot help you with your MAC and the software but if you need an adapter for your Fuji S602Z and a microscope I can help you. We have adapters for all kind of digital cameras either for C-Mount or eye-pieces. Let me know if you want to know more about it.
Unfortunately our web site is not in english yet. However you can find the list of digital cameras we support here:
www.klughammer.de - enter the german pages, then open "Kameraadapter" - "für dig. Kameras" - go to the bottom of this page there you find "Kameraübersicht (PDF)", open it and then you get an overview of cameras.
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bwoAAM} Email: faj-at-highway1.com.au bwoAAM} Name: Faye Taylor
bwoAAM} Organization: Amateur
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bwoAAM} Question: Hello there, bwoAAM} I am a starter who wishes to get her grandchildren interested in a bwoAAM} world beyond TV & computer games. I started using computers when I bwoAAM} purchased my first 128K Mac back in 1985 . My present Mac is a G3 bwoAAM} 192MB ram & 40 GB hard drive.
bwoAAM} I recently aquired a second hand bwoAAM} "MOTIC Biological Series B1 223A " but I have found that I cannot bwoAAM} use my lovely Fuji S602Z digital camera to take photos.
bwoAAM} Do you have any ideas which will enable me to combine the use of the bwoAAM} hardware that I possess? bwoAAM} I feel that the hardest part is getting software that will enable me bwoAAM} to join up to the Macintosh even if I purchased a new camera.
bwoAAM} I would really like to take the photos digitally but is it impossible bwoAAM} with my present configuration? bwoAAM} i would appreciate any comments please
Dear Jon, Caroline sent this to me; please pass it along to your colleague.
Begin forwarded message:
} Bill - } } Kent gave my old machine to a woman in SF who is running it for hire; } I don't know anything about her or the quality of her work. Look at } www.nanoanalytical.netfirms.com . } } Caroline } } -- } Caroline Schooley } Project MICRO Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ } 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 Fri Jan 9 14:49:55 2004
Question: The lab where I currently work uses Scion Image, a freeing distributed graphical editing program. I have been having a problem with the "Set Scale" option under the "Analyze" tab. After taking a snapshot of a known scale under the microscope, I set the scale accordingly by typing in the known distance and setting the units to micrometers. When I switch to a different image and wish to use the same scale, the scale I have just calibrated has been reset to the default. Also, I have gone under the "file" tab and clicked on "record preferences," which seems to do nothing. No save box opens, and I am left with my cursor. In addition, the "revert to save option," also under the "file tab" is never illuminated.
How can I set the scale so it will be calibrated for all images open in the editing session?
Thanks and I hope someone out there has some answers.
I am trying to help a plant biology student with some plant histology on mango saplings. She is interested in looking at paraffin sections of the woody stems (LM). Does anyone have suggestions for a processing schedule? I have my processor set up for human tissues but I could easily extend the programmes to accomodate the cellular nature of the material. Having some suggestions would greatly cut down my trial and error!
Many thanks,
Evelyn Kaplan, Dept of Pathology, College of Medicine and Health Sciences, Sultan Qaboos University, Oman
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 10 00:57:39 2004
Dear friends, the Italian Master on Microscopy at University I Level is starting on February 2nd, 2004. Only few positions are still available due to a delayed registration on January 19th, 2004. Details can be found at "Master Universitario di I livello in Microscopie ed Analisi Microscopiche in Biologia" http://www.studenti.unige.it/corsi/master.html and at www.lambs.it. All my best Alby
p.s. for further information, please e-mail to diaspro-at-fisica.unige.it using "microscopy master 2004" as subject.
....................................................................... ................................... .. non basta, resistere, resistere...resistere ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Alberto Diaspro Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa, Italy voice: +39-0103536426/480 fax 010314218 diaspro-at-fisica.unige.it, http://www.lambs.it http://wiley.com/cda/product/0,,0471409200,00.html
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 11 10:11:10 2004
} } } } Does anyone have a copy of the book "The Cell", 2nd Edition, by Don } } Wayne Fawcett, MD that they are willing to cell. This is a classic book } } full of electron micrographs. I am a graduate student and I will be } } taking an electron microscopy lab this semester and I am looking for 1 } } or more copies of this book. } } } } Please reply to hiswayt-at-earthlink.net } } } } Thank you.
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 11 10:33:04 2004
We occasionally embed plant samples for LM and make a few changes over what is customary for animal tissue. First of all plant samples do not get as brittle if dehydrated in a TBA (tertiary butyl alcohol)/ETOH series ending with 100% TBA.
Start with 15 min in each of 25 and 50% ETOH. Dehydrate for ~ 2-4hrs in each of the following percents: 90 ETOH - 10 TBA 80 ETOH - 20 TBA 65 ETOH - 35 TBA 45 ETOH - 55 TBA 25 ETOH -75 TBA 100% TBA - 3 changes for at least 4hrs total time
Infiltration is helped along by the following: Put samples into an oven set at a sufficiently high temperature to melt your paraffin. (I put all the cassettes into a beaker large enough to hold them so they are completely covered by TBA and then add room for the paraffin. Add solid paraffin (paraplast) to container and allowing it to gradually melt and mix with TBA. The TBA gradually evaporated. The paraplast is then changed a total of 3x over a period of a couple of days prior to embedding tissue in molds.
It is also advisable to use subbed slides or slides coated with poly L-lysine (100µg/ml in 10mM tris at pH 8.0). Plant cell walls tend to lift from the slide surface during staining.
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 1/10/04 12:55 AM, "Evelyn Kaplan" {ekaplan-at-squ.edu.om} 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 } ---------------------------------------------------------------------------- --} - } } } } Good morning, } } I am trying to help a plant biology student with some plant histology on } mango saplings. She is interested in looking at paraffin sections of the } woody stems (LM). Does anyone have suggestions for a processing schedule? I } have my processor set up for human tissues but I could easily extend the } programmes to accomodate the cellular nature of the material. Having some } suggestions would greatly cut down my trial and error! } } Many thanks, } } Evelyn Kaplan, } Dept of Pathology, } College of Medicine and Health Sciences, } Sultan Qaboos University, } Oman } } }
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 11 12:40:08 2004
} } } } } Does anyone have a copy of the book "The Cell", 2nd Edition, by Don } } } Wayne Fawcett, MD that they are willing to cell. This is a classic book } } } full of electron micrographs. I am a graduate student and I will be } } } taking an electron microscopy lab this semester and I am looking for 1 } } } or more copies of this book. } } } } } } Please reply to hiswayt-at-earthlink.net } } } } } } Thank you.
If you look at the used book search site www.abebooks.com you'll find 10 copies at prices from $9-$75. -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 09:54:06 2004
I have used these old protocols with success on woody materials. Put plant material always requires greatly extended times compared to animal tissue. Also, if you have problems with tearing of the embedded tissue during sectioning you can soften the embedded material in Gifford's solution (below). The difficulty is trying to get the harder tissue soft enough so it doesn't tear the softer tissue during sectioning.
The details we used are as follows: Fixation for 12 to 48 hours in FAA (you might also try Navashin's fixative which contains chromic acid) Infiltrate through TBA series (Johansen series) for 2 hours each step 3 changes of pure TBA over 24 hours 3 changes of Paraplast over 24 hours Embed
Soften the blocks by soaking overnight to 7 days in water at up to 38°C If this doesn't work try 2 days to 2 weeks in Gifford's (room temp in fume hood): 20 ml glacial acetic acid 80 ml 60% ethanol 5 ml glycerine You will have to try the softening times with your own tissue. If you leave it too long the soft tissue will become macerated. Let me know if you need more detail.
Good luck,
Kim
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 10:00:07 2004
We have recently acquired a Pelco Biowave and are in the process of acquainting ourselves with it. Currently, I am trying to fix two species of insects with it for SEM (Colorado potato beetle larvae and Diamond back moth larvae). A literature search has not turned up much information on microwave processing for insects. I have tried adaptations of the standard protocol (2% glut, rinse, 1% osmium, rinse, ethanol dehydration, critical point dry) at various microwave wattages, times, and with vacuum applied. So far, I have not achieved reproducible results for either insect. While I plan on more trial and error, I was wondering if anyone has a microwave protocol for insects or any advice on this.
Shannan Little Research Technician/Technicien de recherche Electron Microscopy and Image Analysis / Microscopie électronique et Analyse d'images Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 403-317-3446 Facsimile/Télécopieur: 403-382-3156 P.O. Box 3000 / CP 3000 Lethbridge, Alberta T1J 4B1 littlesm-at-em.agr.ca http://res2.agr.ca/lethbridge/emia/index_e.htm
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 10:00:47 2004
Dear all, We would be interested in users comments about EBSD systems. In particular, we are looking at the systems offered by HKL and by TSL/EDAX. Please comment on some of the following points:
* Ease of use
* Robustness/reliability of indexing when dealing with low symmetry structures
* Calculation of GB misorientations and display of crystallographically equivalent misorientations
* Possibilities for generating different types of map (e.g. orientation, GB misorientation, phase)
* Correlation with EDXS data or maps / generation of combined maps
I look forward to hearing from you. Please copy all responses to myself and to Martin Lee {m.lee-at-earthsci.gla.ac.uk}
Thanks and best wishes
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 10:32:46 2004
I have been calibrating my recently installed Philips TEM with a grating replica and I need some suggestions. At a print magnification of about 80KX I see about a 1% variation in my calculated magnification depending where I select my stop and start marks.
How much variation should I expect in magnification due to changes in lens voltage and current?
Thanks!!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 10:46:04 2004
Frank, I suspect that these variations are in the grating itself, not your TEM. They can be a bit variable, depending on stretching and buckling from the preparation process. We have a record of measurements of semiconductor standard samples going back 8 years on our Jeol 120CX TEM, and find a reproducibility better than 1% over this time. There is a change in magnification from the centre to the edge of the micrographs on our machine of about 1%, but our microscope is now pretty ancient and I would hope that newer machines are much better than this. We are about to embark on a full gauge capability test on the machine, which should be interesting. I can let you know the results of the study if you like.
-----Original Message----- } From: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] Sent: 12 January 2004 16:18 To: microscopy-at-msa.microscopy.com
I have been calibrating my recently installed Philips TEM with a grating replica and I need some suggestions. At a print magnification of about 80KX I see about a 1% variation in my calculated magnification depending where I select my stop and start marks.
How much variation should I expect in magnification due to changes in lens voltage and current?
Thanks!!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 11:13:40 2004
In my experience, successful microwave fixation depends mainly upon the size of the specimen. How big are these insects? Second, if the larvae are difficult to penetrate (I have no idea), longer times may be required. You might also consider Karnofsky's fixative instead of Glutaraldehyde (I found that it improved results in many cases over a wide variety of specimen types, although I didn't try insects). I would try lengthening the primary fixation time before adjusting any of the other processing variables. Fixation temperatures should never exceed 50°C (I don't know what this translates to in watts on your Biowave), or you will get "crispy critters".
best regards, Steven Slap Microwave Consultant
At 11:04 AM -0500 1/12/04, Shannan Little wrote: } We have recently acquired a Pelco Biowave and are in the process of } acquainting ourselves with it. Currently, I am trying to fix two species } of insects with it for SEM (Colorado potato beetle larvae and Diamond } back moth larvae). A literature search has not turned up much } information on microwave processing for insects. I have tried } adaptations of the standard protocol (2% glut, rinse, 1% osmium, rinse, } ethanol dehydration, critical point dry) at various microwave wattages, } times, and with vacuum applied. So far, I have not achieved reproducible } results for either insect. While I plan on more trial and error, I was } wondering if anyone has a microwave protocol for insects or any advice } on this.
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 11:46:35 2004
FYI I just opened a new supply of Kodak Professional Rapid Fixer and found larger boxes. After the film switch not so very long ago, I decided to check the ingredients. "Solution A" now has Ammonium Sulfite, Sodium bisulfite and Sodium acetate added to what was printed on the old box. The mixing directions are the same 1999 version. "Solution B" and the CAT # 146 4106 appear to be the same.
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145 psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 11:53:21 2004
We are trying to install two cameras on a Windows XP computer.
We cannot get XP to recognize both the Sensicam PCI card and the Roper PCI card. It's completely an exclsuive or installation. We've poked around the Device driver menu and downloaded the most recent drivers.
Has anybody figured out how to install both these camera simultaneously?
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 12:01:24 2004
I would recommend the Mag-I-Cal sample for calibrating your TEM. You can do a much larger range of Magnification settings than using a replica. I think that the error is 1 to 2%. In addition, you can do rotation and camera constant calibrations with the same sample.
One thing to minimize the variation in the microscope is to always changed the lenses in the same direction to avoid hysterisis and to make sure that you are at the eucentric point so that the obj lens is the same. Again, you should bring the focus from the same direction.
If the goniometer has not bee removed or the column split, the values that you get from time to time should be very close. We had to run calibration twice a year on the TEM. I can't remember the range specifically, but I think that the variation in mag was less than 0.5% and may have been better than 0.1% when the above criteria was met.
-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: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] Sent: Monday, January 12, 2004 11:18 AM To: microscopy-at-msa.microscopy.com
I have been calibrating my recently installed Philips TEM with a grating replica and I need some suggestions. At a print magnification of about 80KX I see about a 1% variation in my calculated magnification depending where I select my stop and start marks.
How much variation should I expect in magnification due to changes in lens voltage and current?
Thanks!!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 12:53:50 2004
When we manufacture TEM or SEM gratings we make them from a replica of a master grating. When you dissolve away the replication material there is some shrinkage, but it should be very limited. The shrinkage is inherent in the manufacturing, but we cull any which show problematic shrinkage. It is very similar to our carbon substrate manufacturing.
John Arnott
Disclaimer: Ladd Research manufactures and sells the gratings, replicating materials and substrates mentioned in this email.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
----- Original Message ----- } From: {Frank.Karl-at-degussa.com} To: {microscopy-at-msa.microscopy.com} Sent: Monday, January 12, 2004 11:17 AM
Frank, If you are using film, a second variation in measurement may come from the enlarger when you print the picture. If the negative is not supported on glass, it can bow in the center and distort the measurement a bit.
It was a surprise to me to find that if I had set my "MAG. ZERO" early in the morning and then checked it later in the day, there was frequently a slight change. It was explained to me that in an old building, when there was a greater draw of electricity, a change could be expected and for really important work, I should re-calibrate. Am I just gullable?
My favorite goof has been the result of not adjusting my tilting specimen holder to read 0 degrees!
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145 psconnel-at-sas.upenn.edu } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } When we manufacture TEM or SEM gratings we make them from a replica of a } master grating. When you dissolve away the replication material there is } some shrinkage, but it should be very limited. The shrinkage is inherent in } the manufacturing, but we cull any which show problematic shrinkage. } It is very similar to our carbon substrate manufacturing. } } John Arnott } } Disclaimer: Ladd Research manufactures and sells the gratings, replicating } materials and substrates mentioned in this email. } } Ladd Research } 83 Holly Court } Williston, VT 05495 } On-line Catalog: http://www.laddresearch.com } tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) } fax: 1-802-660-8859 } e-mail: sales-at-laddresearch.com } } ----- Original Message ----- } } From: {Frank.Karl-at-degussa.com} } To: {microscopy-at-msa.microscopy.com} } Sent: Monday, January 12, 2004 11:17 AM } Subject: [Microscopy] TEM mag question } ----------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } I have been calibrating my recently installed Philips TEM with a grating } } replica and I need some suggestions. At a print magnification of about } } 80KX I see about a 1% variation in my calculated magnification depending } } where I select my stop and start marks. } } } } How much variation should I expect in magnification due to changes in lens } } voltage and current? } } } Frank Karl
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 15:39:11 2004
I wanted to know the vapor pressure of alumina (Al2O3) at 1200 C. I know that it will be very low, but an estimate would also be helpful. I looked at various places (such as CRC handbook of Physics and Chemistry, Handbook of thermophysical properties of solid materials, ASM handbook, Vol. 5), but got values above 1950C (e.g. 10^-3 torr at 1950 C).
I want to know the answer to this question since I work at vacuum levels of 10^-6 to 10^-8 torr and at 1200C. I am interested in knowing if the aluminum oxide layer on my samples evaporates under these conditions. One company representative said that it wont, but he did not have vapor pressure values to support the assertion.
thanks in advance
Rahul Panat Univ of Illinois Urbana, IL
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 17:23:47 2004
Title-Subject: [Microscopy] [Filtered] Kodak 4489 EM film
Question: Can anyone use some left-over Kodak 4489 electron microscopy film? I was about to throw out 5 boxes [2 x 10 in. 100 strips per box] during "housecleaning" the other day, but thought there might be someone who could use it. It has been refrigerated and unopened for about 12 years. [I used to use it in our old RCA EMU 3G TEM.] If someone wants it send me your mailing address and I'll ship it out to you. Otherwise, it will be recycled or trashed.
Lee Hadden Department of Biology Wingate University Wingate, NC 28174
Question: I am interested in getting quantitative analysis training for analyzing aluminum alloys and oxides. In our lab, we have an Hitachi S-3500N equipped with an Oxford EDS Spectrometer (model# 7021)that has the INCA and ISIS software packages installed on the computer. I would like to visit an independent laboratory or university for a week or two for training. If someone could assist me I would appreciate it.
} } Hello Shannan. } } I have some experience with microwave fixation of Drosophila larval } salivary glands for TEM. First, I run my cold spot -at- 15-18 degrees (this } removes the heating effect) I fix them in Karnovsky's fix power level } 1(100 watts on my MW) for 1 minute on-1minute off-1 minute on). Then I } turn the power up to 450 watts (power level 4 on my machine) and pulse for } 30 seconds on-30 seconds off-30 seconds on 3 times.They should not get } hot! I let sit on the bench in fixative for 5 minutes. I have also tried } this protocol on zebrafish larvae (with vacuum) and they are well fixed. } The insect probably has a cuticle which may hinder the penetration of the } fixative. If you can find a way to partially remove this, or inject the } fix then MW, you might have better results. } For osmium fix, I repeat the above timetable. I dehydrate under vacuum 45 } seconds -at- power level 3(350 watts) 50, 70,95% once each. I do 100% } ethanol 3 times followed by 100% acetone before infiltration in resin. } Hope this helps, JoAnn Buchanan } } } } We have recently acquired a Pelco Biowave and are in the process of } } acquainting ourselves with it. Currently, I am trying to fix two species } } of insects with it for SEM (Colorado potato beetle larvae and Diamond } } back moth larvae). A literature search has not turned up much } } information on microwave processing for insects. I have tried } } adaptations of the standard protocol (2% glut, rinse, 1% osmium, rinse, } } ethanol dehydration, critical point dry) at various microwave wattages, } } times, and with vacuum applied. So far, I have not achieved reproducible } } results for either insect. While I plan on more trial and error, I was } } wondering if anyone has a microwave protocol for insects or any advice } } on this.
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 19:07:09 2004
Michael, Have you tried rearranging the cards on the Mobo? I had the same adventure when installing a Pixera camera and Scion system on an XP system. I did not get the New Hardware Found announcement for the Pixera card until I did some card swapping.
Let me know how you get on.
Greg
Dr. G. F. Barclay Plant Science Unit, Dept of Life Sciences University of the West Indies St. Augustine, Trinidad and Tobago West Indies Phone: 868 645 3232 ext 3112/2045 Fax: 868 645 7132
} From: Michael Cammer {cammer-at-aecom.yu.edu} } To: microscopy-at-MSA.microscopy.com } Subject: [Microscopy] Sensicam QE & Roper HQ } Date: Mon, 12 Jan 2004 12:58:55 -0500 } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_________________________________________________________________ Help STOP SPAM with the new MSN 8 and get 2 months FREE* http://join.msn.com/?page=features/junkmail
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 20:02:39 2004
Dear Shannan, Below is a method we have used for a few different insect types, for TEM not SEM, but the fixing steps we use might provide a useful comparison at least.
With SEM I assume you won't want to dissect your samples prior to processing (as we did) so penetration of the solutions may be more of a problem, but then the requirements for fixation for SEM are also less stringent. The microwave conditions may be useful guidelines but of course you'll have to determine the conditions for your own microwave and samples. If you haven't already got them, I strongly recommend Gary Login's text (Login, G. R., & Dvorak, A. M. (1994) " The Microwave Toolbook A Practical Guide for Microscopist" Boston: Beth Israel Hospital) and Kok and Boon's book "The Microwave Cookbook for Microscopists", Boon and Kok, Coulomb Press Leiden, 1992.
Method: Primary fix: 3% glutaraldehyde in 0.1M Na cacodylate, pH 7.4. Samples dissected out and placed into the primary fix solution (in a small plastic petri dish).
They were then put into fresh fixative (specimen containers for a Leica AFS were used inside the petri dish) and microwave irradiated as follows:
EMS lab microwave oven setup: -a dummy load of 100ml dw at 20degC in a 250 glass beaker was used (always in the same place - right rear corner for us) -temperature limiting off -100% 'power' (ie magnetron on continuously) -sample volume for all the fixing steps was 4.0ml -magnetron was pre-warmed for 2 minutes with a load of 500-600ml water before each step (unless the oven was used less than 2 minutes earlier).
1) Microwave Primary Fixation:
The sample in 4 ml of primary fix is irradiated for (7s) to give a final temperature of about 50degC - check the temperature after irradiation (and obviously before you use your actual samples if you're doing this for the first time) and alter subsequent run times if necessary. (We use the spot in the oven we deem to be receiving a steady, high level of radiation).
Allow sample to sit in fume cupboard in fix container for 3 minutes to cool it to room temp before removing fix. Replace fix with fresh and repeat the irradiation process twice. A cool dummy load must used with each run.
Leave samples in fume cupboard for 30 minutes at room temperature.
2) Rinse the samples:
Three X 10 minutes in 0.1M cacodylate buffer.
3) Secondary fixation:
Place 2% OsO4 in 0.1M cacodylate buffer into fix dish with specimens. Irradiate for 7s Leave for 40 minutes in the OsO4.
4) Rinse with 0.1M cacodylate buffer
Three X 10 minutes
The remainder of my method is for TEM preparation so you could do your usual pre-drying and drying steps then.
Regards,
Richard
} } We have recently acquired a Pelco Biowave and are in the process of } acquainting ourselves with it. Currently, I am trying to fix two species } of insects with it for SEM (Colorado potato beetle larvae and Diamond } back moth larvae). A literature search has not turned up much } information on microwave processing for insects. I have tried } adaptations of the standard protocol (2% glut, rinse, 1% osmium, rinse, } ethanol dehydration, critical point dry) at various microwave wattages, } times, and with vacuum applied. So far, I have not achieved reproducible } results for either insect. While I plan on more trial and error, I was } wondering if anyone has a microwave protocol for insects or any advice } on this. } } Shannan Little
Richard Easingwood Otago Centre for Electron Microscopy Department of Anatomy and Structural Biology School of Medical Sciences, University of Otago PO Box 913, Dunedin, NEW ZEALAND Telephone: 0064 3 479 7301 Facsimile: 0064 3 479 7254 GSM: 0064 21 222 4759 mailto:richard.easingwood-at-stonebow.otago.ac.nz Web site: http://ocem.otago.ac.nz/
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 21:28:45 2004
We swapped cards all over the place. The one in the lowest slot gets recognized, but not the other. Also, the Roper card doesn't seem to work in the highest slot, even alone. We could get either card to work with a firewire camera in another slot, but the Retiga we have isn't low noise enough for this application. I think we'll have to temporarily get a second computer in the room. Thanks.
On Tue, 13 Jan 2004, gregor barclay wrote:
} Michael, } Have you tried rearranging the cards on the Mobo? I had the same adventure } when installing a Pixera camera and Scion system on an XP system. I did not } get the New Hardware Found announcement for the Pixera card until I did some } card swapping. } } Let me know how you get on. } } Greg } } } } Dr. G. F. Barclay } Plant Science Unit, Dept of Life Sciences } University of the West Indies } St. Augustine, Trinidad and Tobago } West Indies } Phone: 868 645 3232 ext 3112/2045 } Fax: 868 645 7132 } } } } } } } From: Michael Cammer {cammer-at-aecom.yu.edu} } } To: microscopy-at-MSA.microscopy.com } } Subject: [Microscopy] Sensicam QE & Roper HQ } } Date: Mon, 12 Jan 2004 12:58:55 -0500 } } } } } } } } ------------------------------------------------------------------------------ } } 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 } } ------------------------------------------------------------------------------- } } } } We are trying to install two cameras on a Windows XP computer. } } } } We cannot get XP to recognize both the Sensicam PCI card and the Roper PCI } } card. It's completely an exclsuive or installation. We've poked around } } the Device driver menu and downloaded the most recent drivers. } } } } Has anybody figured out how to install both these camera simultaneously? } } } } } } } } } } ____________________________________________________________________________ } } Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of } } Med. } } Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 } } (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ } } } } } } _________________________________________________________________ } Help STOP SPAM with the new MSN 8 and get 2 months FREE* } http://join.msn.com/?page=features/junkmail } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 22:57:10 2004
We have an old Wild M8 dissecting microscope with a 1.0x objective that can be converted to 0.4x or 1.6x by clipping adapter lenses onto the bottom of the 1.0x objective. We'd like to get a second set for another M8 because they get used so much now that they are sometimes needed by both microscopes. Unfortunately, these are no longer available from Leica, instead you have to get an adapter and rather more expensive 0.4x and 1.6x objectives. These new objectives are better quality than the old adapters, but for our work, the ease of switching with the adapters and low cost outweighs the marginal increase in quality at the magnifications we're using.
Anyone willing to part with their adapters, or know of a source? I've tried ebay and several other used equipment sites with no luck so far.
0.4x adapter lens part no. 367898 1.6x adapter lens part no. 367916
Thanks much, Rosemary White
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre fax 61- 2 6246 5000 CSIRO Plant Industry ph. 61- 2 6246 5475 or GPO Box 1600 mob. 61- 0402 835 973 Canberra, ACT 2601, Australia
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 13 07:23:25 2004
Richard makes some good points here. I second his reading recommendations, and want to let everyone know that there is a brand new edition of Kok and Boon, Microwaves for the Art of Microscopy, Coulomb Press, Leiden, 2003, which has some very useful new information.
I believe that the use of a dummy load is not needed in the Biowave, which has a recirculating water cooler. In any event, the end temperature of 50°C is the key, and Richard's 3 x 7 minutes should give Shannon a good starting point for time in the fixative. I would be a little concerned about using such a small volume of fixative as Richard did (4.0 ml), both because of a fear of exceeding the temperature set point and because a greater volume of fixative to specimen ratio seems prudent based upon my experience with light microscopy specimens. I generally work with about 11.0 ml of fixative.
best regards, Steven Slap Microwave Consultant
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 13 07:44:35 2004
O.k., folks following the various recommendations from the list for flatbed scanners we got an Microtek AtrixScan 2500f.
So now, anyone out there with a Microtek 2500f what is the part number and where do you get the bulbs from?
Microtek offical position is "There are no user servicable parts. You have to ship it back - at your cost - to Microtek in California for repair". Now, I am not shipping a 100lb scanner back to California for a $20 bulb replacment - let alone doing it every 6-8 months. Surely someone else out there has already come across this (especially since the bulbs never power down).
Thank you in advance for any help.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 13 08:21:32 2004
Question: I am interested in getting quantitative analysis training for analyzing aluminum alloys and oxides. In our lab, we have an Hitachi S-3500N equipped with an Oxford EDS Spectrometer (model# 7021)that has the INCA and ISIS software packages installed on the computer. I would like to visit an independent laboratory or university for a week or two for training. If someone could assist me I would appreciate it.
My tables* go down to 10^-6, and Al2O3 reaches that vapor pressure at 1637C. I expect that the vapor pressure would be well below your vacuum level at 1200C.
*The Characterization of High Temperature Vapors, J. L. Margrave, Ed. John Wiley, 1967.
John Friel
-- --------------------------- John J. Friel Princeton Gamma-Tech 1026 Rte. 518 Rocky Hill, NJ 08553 (609) 924-7310 x232 phone (609) 924-1729 fax E-mail: jjf-at-pgt.com Web page: www.pgt.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 13 09:51:03 2004
I can't give you definite answer, but a couple of hints that may help you:
1) Some PCs sense the existence of cards in the slots and turn off the slots if no card is present. Perhaps the higher slots get turned off. Check in the BIOS if the computer is in some kind of power saving mode.
2) Take it one at a time. Install one card and get it to work. then take that card out and install the other card in another slot. If you get that to work, put the first back in and install again.
3) If one card is not recorgnized, check if there are any conflicts in the device manager.
4) If one card does not work at all, contact the manufacturer and get their help.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Michael Cammer [mailto:cammer-at-aecom.yu.edu] Sent: Monday, January 12, 2004 20:34 To: gregor barclay Cc: microscopy-at-MSA.microscopy.com
We swapped cards all over the place. The one in the lowest slot gets recognized, but not the other. Also, the Roper card doesn't seem to work in the highest slot, even alone. We could get either card to work with a firewire camera in another slot, but the Retiga we have isn't low noise enough for this application. I think we'll have to temporarily get a second computer in the room. Thanks.
On Tue, 13 Jan 2004, gregor barclay wrote:
} Michael, } Have you tried rearranging the cards on the Mobo? I had the same adventure
} when installing a Pixera camera and Scion system on an XP system. I did not } get the New Hardware Found announcement for the Pixera card until I did some } card swapping. } } Let me know how you get on. } } Greg } } } } Dr. G. F. Barclay } Plant Science Unit, Dept of Life Sciences } University of the West Indies } St. Augustine, Trinidad and Tobago } West Indies } Phone: 868 645 3232 ext 3112/2045 } Fax: 868 645 7132 } } } } } } } From: Michael Cammer {cammer-at-aecom.yu.edu} } } To: microscopy-at-MSA.microscopy.com } } Subject: [Microscopy] Sensicam QE & Roper HQ } } Date: Mon, 12 Jan 2004 12:58:55 -0500 } } } } } } } } --------------------------------------------------------------------------- --- } } 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 } } --------------------------------------------------------------------------- ---- } } } } We are trying to install two cameras on a Windows XP computer. } } } } We cannot get XP to recognize both the Sensicam PCI card and the Roper PCI } } card. It's completely an exclsuive or installation. We've poked around } } the Device driver menu and downloaded the most recent drivers. } } } } Has anybody figured out how to install both these camera simultaneously? } } } } } } } } } } ___________________________________________________________________________ _ } } Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of } } Med. } } Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 } } (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ } } } } } } _________________________________________________________________ } Help STOP SPAM with the new MSN 8 and get 2 months FREE* } http://join.msn.com/?page=features/junkmail } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 13 10:55:59 2004
} 3) If one card is not recorgnized, check if there are } any conflicts in the device manager.
It's difficult for me to add anything to Mike's response, but to note that many motherboards inherantly share resources amongst pairs of PCI slots. For example, many mobos share the resources of AGP slot with the next PCI slot. Another observation of note is that the Windows OS is designed for sharing resources amongst slots such there should be no conflicts, but my own experience is how well this works is highly dependent on the manufacturer's software driver for the card.
Therefore, I'd suggest you may not yet have found the right combination of slots, or that you beg the manufacturer(s) for their help.
} -----Original Message----- } } From: Michael Cammer [mailto:cammer-at-aecom.yu.edu] } Sent: Monday, January 12, 2004 20:34 } To: gregor barclay } Cc: microscopy-at-MSA.microscopy.com } Subject: [Microscopy] RE: Sensicam QE & Roper HQ } } } } } ------------------------------------------------------------------ } ---------- } -- } 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 } ------------------------------------------------------------------ } ---------- } --- } } We swapped cards all over the place. The one in the lowest slot gets } recognized, but not the other. Also, the Roper card doesn't seem to } work in the highest slot, even alone. We could get either card to work } with a firewire camera in another slot, but the Retiga we have isn't low } noise enough for this application. } I think we'll have to temporarily get a second computer in the room. } Thanks. } } On Tue, 13 Jan 2004, gregor barclay wrote: } } } Michael, } } Have you tried rearranging the cards on the Mobo? I had the } same adventure } } } when installing a Pixera camera and Scion system on an XP system. I did } not } } get the New Hardware Found announcement for the Pixera card until I did } some } } card swapping. } } } } Let me know how you get on. } } } } Greg } } } } } } } } Dr. G. F. Barclay } } Plant Science Unit, Dept of Life Sciences } } University of the West Indies } } St. Augustine, Trinidad and Tobago } } West Indies } } Phone: 868 645 3232 ext 3112/2045 } } Fax: 868 645 7132 } } } } } } } } } } } } } From: Michael Cammer {cammer-at-aecom.yu.edu} } } } To: microscopy-at-MSA.microscopy.com } } } Subject: [Microscopy] Sensicam QE & Roper HQ } } } Date: Mon, 12 Jan 2004 12:58:55 -0500 } } } } } } } } } } } } } ----------------------------------------------------------------- } ---------- } --- } } } 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 } } } } ----------------------------------------------------------------- } ---------- } ---- } } } } } } We are trying to install two cameras on a Windows XP computer. } } } } } } We cannot get XP to recognize both the Sensicam PCI card and the Roper } PCI } } } card. It's completely an exclsuive or installation. We've } poked around } } } the Device driver menu and downloaded the most recent drivers. } } } } } } Has anybody figured out how to install both these camera } simultaneously? } } } } } } } } } } } } } } } } _________________________________________________________________ __________ } _ } } } Michael Cammer Analytical Imaging Facility Albert Einstein } Coll. of } } } Med. } } } Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY } 10461 } } } (718) 430-2890 Fax: 430-8996 URL: } http://www.aecom.yu.edu/aif/ } } } } } } } } } } _________________________________________________________________ } } Help STOP SPAM with the new MSN 8 and get 2 months FREE* } } http://join.msn.com/?page=features/junkmail } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 13 12:17:42 2004
This discussion on fixation is interesting from a number of aspects. The reliance on answers based on almost 10 year old literature and technology, without consideration of current literature, technique and technology (PELCO BioWave) is curious. In the last three years at the Microscopy and Microanalysis meetings whole and half day sessions have been devoted to microwave processing techniques using new and emerging technology. Current experimental findings do not support the old literature especially when it comes to fixation and microwave heating. If one chose to stay reasonably current with both the new technology and techniques they would not propose 50C as an end point temperature for the fixative. I would direct those curious to a recent paper in Ultrastructural Pathology (2003) 27:187-196, Microwave and Digital Imaging Technology Reduce Turnaround Times for Diagnostic Electron Microscopy and as well as Microwave Techniques and Protocols (2001) from Humana Press, Totowa, NJ and chapter 16 in particular. I have been around the microwave scene for over 10 years and have worked diligently to improve the technology (the dreaded vendor) as well as the science (proof side of the equation). I have difficulty with advice that is 10 years old and ignores recent developments.
Richard T. Giberson Manager Research and Development Ted Pella, Inc.
-----Original Message----- } From: Steven E. Slap [mailto:siksik03-at-comcast.net] Sent: Tuesday, January 13, 2004 5:28 AM To: Richard Easingwood Cc: microscopy-at-msa.microscopy.com
Dear fellow microscopists
Richard makes some good points here. I second his reading recommendations, and want to let everyone know that there is a brand new edition of Kok and Boon, Microwaves for the Art of Microscopy, Coulomb Press, Leiden, 2003, which has some very useful new information.
I believe that the use of a dummy load is not needed in the Biowave, which has a recirculating water cooler. In any event, the end temperature of 50°C is the key, and Richard's 3 x 7 minutes should give Shannon a good starting point for time in the fixative. I would be a little concerned about using such a small volume of fixative as Richard did (4.0 ml), both because of a fear of exceeding the temperature set point and because a greater volume of fixative to specimen ratio seems prudent based upon my experience with light microscopy specimens. I generally work with about 11.0 ml of fixative.
best regards, Steven Slap Microwave Consultant
} --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 13 18:13:26 2004
Organization: Fred Hutchinson Cancer Research Center
Title-Subject: [Microscopy] [Filtered] immuno scanning electron microscopy
Question: We would appreciate some help if anyone has knowledge of or has used a method for labelling tissue of cell walls-stomach or intestine-with ICam-1 for immuno scanning E.M.
We are interested in using image analysis software for several purposes: 1. to speed the process of counting and measuring of cells in unialgal cultures 2. to assist in counting cells in simple two species competition experiments where they are relatively easy to separate by shape. 3. to develop the capacity to identify and count the dominant species from natural samples
To this end we have been offered some software packages with an epifluorescent inverted microscope. We would like to know if anyone has any experience with these software packages trying to achieve similar goals.
1. Image Pro Discovery (by Media Cybernetics) 2. Softimaging System (SIS) Auto version 3.2 4. Leica QWin Standard software 5. Axiovision 4 (Zeiss) 6. Digital Optics V++ Image analysis software
Comments on your experiences would be most welcome.
Many Thanks and All the best for the coming New Year Leanne Armand and Peter Thompson
___________________________________________ *Contact Days - Tues., Wed., Fri.* ___________________________________________ Dr Leanne Armand Joint Personal Assistant to Dr Peter Thompson Sustainable Marine Ecosystems in the South East CSIRO Marine Research GPO Box 1538 Hobart, 7001 Tas. Australia
Title-Subject: [Microscopy] [Filtered] SEM: Minimum Vacuum for Tungsten Filament
Question: We are currently experiencing vacuum issues which are causing tungsten oxide growth and premature failure of the filament while operating our JEOL 5900. One of the JEOL technicians has suggested that 5x10-4 torr is acceptable, but I find this hard to believe. The issue we are dealing with is extraordinarily slow pumping times -- 1 hour to obtain 3x10-5 torr and a max vacuum of 5x10-6 torr after 3-4 hours. We are a high volume lab, and so I wish to know what the minimum vacuum I need without seriously diminishing filament life, in order to maximize the workload.
Sincerely,
Cavin T. F. Mooers EM Facility Manager Vitreous State Laboratory The Catholic University of America Hannan Hall ñ Rm 433 Washington, D.C. 20064 (202) 319-6237 (Office) (202) 319-5346 (Lab) (202) 319-4469 (Fax)
Email: ann-at-northwestern.edu Name: Ann Chiaramonti
Organization: Northwestern University
Title-Subject: [Microscopy] [Filtered] MListserver: Teaching samples for TEM
Question: Hi, I am teaching a TEM lab class this quarter and am looking for a material useful for teaching basic imaging and diffraction in the TEM. We would like a material with lots of dislocations, stacking faults, grain boundaries, etc. so that it is interesting for the students. I would like to use stainless steel but do not have any non-magnetic readily available. I would like to avoid using perchloric acid if possible in the preparation. Any suggestions/comments? Thank you in advance,
Ann Chiaramonti Northwestern University ann-at-northwestern.edu
What's the latest word on glow discharge for grids?
We have an old VE that will glow discharge, but have hardly ever used it. A new faculty person wants to do more glow discharging and is looking for info.
Somewhere I have a paper describing a home made glow discharge device, anybody know about this and if it works?
Also have heard that keeping grids in the refrigerator helps too. What's up with that?
Her application is carbon films for negatively stained macromolecules.
Thanks
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 Jan 14 18:24:33 2004
Dear Cavin, It sounds like the problem is not just that you have a leak, but where the leak is. The vacuum is usually tested at the top of the diffusion pump, but what matters for the filament is the vacuum at the gun. If you have a small leak at the gun, the vacuum near the filament may be quite a bit worse than you are seeing. I routinely turn on the tungsten filament when the vacuum reads 1X10-4 torr, but I am sure that that vacuum is consistent in the column and that it will rapidly improve. I suggest you assume that the leak is near the gun and do some detective work. You will be much happier, the filament will last longer and your throughput will better. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "by way of MicroscopyListserver" {cavinm-at-vsl.cua.edu} To: {microscopy-at-ns.microscopy.com} Sent: Wednesday, January 14, 2004 2:56 PM
Title-Subject: [Microscopy] [Filtered] SEM: Minimum Vacuum for Tungsten Filament
Question: We are currently experiencing vacuum issues which are causing tungsten oxide growth and premature failure of the filament while operating our JEOL 5900. One of the JEOL technicians has suggested that 5x10-4 torr is acceptable, but I find this hard to believe. The issue we are dealing with is extraordinarily slow pumping times -- 1 hour to obtain 3x10-5 torr and a max vacuum of 5x10-6 torr after 3-4 hours. We are a high volume lab, and so I wish to know what the minimum vacuum I need without seriously diminishing filament life, in order to maximize the workload.
Sincerely,
Cavin T. F. Mooers EM Facility Manager Vitreous State Laboratory The Catholic University of America Hannan Hall ñ Rm 433 Washington, D.C. 20064 (202) 319-6237 (Office) (202) 319-5346 (Lab) (202) 319-4469 (Fax)
Thanks to Richard Giberson for his comments. As Pelco is not represented in this part of the world, I was unaware of this advance in microwave technology and the new Pelco BioWave.
We have a national microscopy conference in February next year (see http://ocem.otago.ac.nz/Conference_2005/Microscopy_NZ_2005.html) It would be fantastic if Pelco would bring this technology out to demonstrate it to us here during the conference!
Regards,
Richard
} Dear Microscopists, } } This discussion on fixation is interesting from a number of aspects. The } reliance on answers based on almost 10 year old literature and technology, } without consideration of current literature, technique and technology (PELCO } BioWave) is curious. In the last three years at the Microscopy and } Microanalysis meetings whole and half day sessions have been devoted to } microwave processing techniques using new and emerging technology. Current } experimental findings do not support the old literature especially when it } comes to fixation and microwave heating. If one chose to stay reasonably } current with both the new technology and techniques they would not propose } 50C as an end point temperature for the fixative. I would direct those } curious to a recent paper in Ultrastructural Pathology (2003) 27:187-196, } Microwave and Digital Imaging Technology Reduce Turnaround Times for } Diagnostic Electron Microscopy and as well as Microwave Techniques and } Protocols (2001) from Humana Press, Totowa, NJ and chapter 16 in particular. } I have been around the microwave scene for over 10 years and have worked } diligently to improve the technology (the dreaded vendor) as well as the } science (proof side of the equation). I have difficulty with advice that is } 10 years old and ignores recent developments. } } Richard T. Giberson } Manager Research and Development } Ted Pella, Inc. } } -----Original Message----- } } From: Steven E. Slap [mailto:siksik03-at-comcast.net] } Sent: Tuesday, January 13, 2004 5:28 AM } To: Richard Easingwood } Cc: microscopy-at-msa.microscopy.com } Subject: [Microscopy] Re: microwave fixation procedure for insects } } }
Richard Easingwood Otago Centre for Electron Microscopy Department of Anatomy and Structural Biology School of Medical Sciences, University of Otago PO Box 913, Dunedin, NEW ZEALAND Telephone: 0064 3 479 7301 Facsimile: 0064 3 479 7254 GSM: 0064 21 222 4759 mailto:richard.easingwood-at-stonebow.otago.ac.nz Web site: http://ocem.otago.ac.nz/
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 14 18:28:58 2004
Dear Ann, Brass, annealed or slightly deformed, is easy to thin in a high concentration nitric acid in methanol electropolishing bath (see Van der Voort). It is non-magnetic, has lots of twins, dislocations and an easy diffraction pattern. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "by way of MicroscopyListserver" {ann-at-northwestern.edu} To: {microscopy-at-ns.microscopy.com} Sent: Wednesday, January 14, 2004 2:57 PM
On Jan 13, 2004, at 5:49 AM, Richard Edelmann wrote:
} So now, anyone out there with a Microtek 2500f what is the part number } and where do you get the bulbs from? } Dear Richard, We have a different model and have yet to need a bulb replaced; however, we purchased the unit from Calumet photo, and I recommend talking to Chris Benes, (323)466-1238 x108, chris.benes-at-calumetphoto.com. (This is in the Pacific Time Zone, so plan accordingly.) Good luck, and please pass along what you learn. 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 Jan 14 21:28:06 2004
Jonathan Krupp wrote: ==================================================== What's the latest word on glow discharge for grids?
We have an old VE that will glow discharge, but have hardly ever used it. A new faculty person wants to do more glow discharging and is looking for info
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 08:04:18 2004
The Pelco BioWave is not the only new advance in microwave technology. For something else really new, check out Milestone's REM microwave for EM processing (http://www.milestonesrl.com). It has a non-contact infra-red temperature sensor, full computer control by touch screen and no need for a water load at all. This is another microwave that you need to have at your conference. Maybe you can get someone from Milestone (JIm Milius?) to present it.
best regards, Steven Slap Microwave Consultant
} } Thanks to Richard Giberson for his comments. As Pelco is not represented in } this part of the world, I was unaware of this advance in microwave } technology and the new Pelco BioWave. } } We have a national microscopy conference in February next year (see } http://ocem.otago.ac.nz/Conference_2005/Microscopy_NZ_2005.html) It would } be fantastic if Pelco would bring this technology out to demonstrate it to } us here during the conference! } } Regards, } } } Richard } } } } } Dear Microscopists, } } } } This discussion on fixation is interesting from a number of aspects. The } } reliance on answers based on almost 10 year old literature and technology, } } without consideration of current literature, technique and technology (PELCO } } BioWave) is curious. In the last three years at the Microscopy and } } Microanalysis meetings whole and half day sessions have been devoted to } } microwave processing techniques using new and emerging technology. Current } } experimental findings do not support the old literature especially when it } } comes to fixation and microwave heating. If one chose to stay reasonably } } current with both the new technology and techniques they would not propose } } 50C as an end point temperature for the fixative. I would direct those } } curious to a recent paper in Ultrastructural Pathology (2003) 27:187-196, } } Microwave and Digital Imaging Technology Reduce Turnaround Times for } } Diagnostic Electron Microscopy and as well as Microwave Techniques and } } Protocols (2001) from Humana Press, Totowa, NJ and chapter 16 in particular. } } I have been around the microwave scene for over 10 years and have worked } } diligently to improve the technology (the dreaded vendor) as well as the } } science (proof side of the equation). I have difficulty with advice that is } } 10 years old and ignores recent developments. } } } } Richard T. Giberson } } Manager Research and Development } } Ted Pella, Inc. } } } } -----Original Message----- } } } From: Steven E. Slap [mailto:siksik03-at-comcast.net] } } Sent: Tuesday, January 13, 2004 5:28 AM } } To: Richard Easingwood } } Cc: microscopy-at-msa.microscopy.com } } Subject: [Microscopy] Re: microwave fixation procedure for insects } } } } } } } } Richard Easingwood } Otago Centre for Electron Microscopy } Department of Anatomy and Structural Biology } School of Medical Sciences, University of Otago } PO Box 913, Dunedin, NEW ZEALAND } Telephone: 0064 3 479 7301 } Facsimile: 0064 3 479 7254 } GSM: 0064 21 222 4759 } mailto:richard.easingwood-at-stonebow.otago.ac.nz } Web site: http://ocem.otago.ac.nz/
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 08:54:58 2004
Yes, 5x10-4 is a useable vacuum, but it will shorten fialment life. As previously suggested you may have a vacuum leak in the gun (check the gun chamber o-ring). But since you are getting down to 5x10-6 it doesn't sound like a leak is the main problem. Becasue it takes so long to pump down, sounds more like a pumping problem. Not exactly sure where your starting from when you say it takes 1 hour to get 3 x10-5. I just vented my JEOL 840 gun and it took ~ 9 mins to get the same with a warmed and running pumping system.
-- dirty or cracked DP oil or RP oil (Change oils)
-- poor DP backing (clean RP and replace RP exhaust filter).
-- low RP oil level or DP oil level.
-- check cooling temp on DP. (Too high *or* too low)
Good luck.
} Email: cavinm-at-vsl.cua.edu } Name: Cavin Mooers } } Organization: Vitreous State Lab } } Title-Subject: [Microscopy] [Filtered] SEM: Minimum Vacuum for Tungsten Filament } } Question: We are currently experiencing vacuum } issues which are causing tungsten oxide growth } and premature failure of the filament while } operating our JEOL 5900. One of the JEOL } technicians has suggested that 5x10-4 torr is } acceptable, but I find this hard to believe. The } issue we are dealing with is extraordinarily slow } pumping times -- 1 hour to obtain 3x10-5 torr and } a max vacuum of 5x10-6 torr after 3-4 hours. We } are a high volume lab, and so I wish to know what } the minimum vacuum I need without seriously } diminishing filament life, in order to maximize } the workload. } } Sincerely, } } Cavin T. F. Mooers } EM Facility Manager } Vitreous State Laboratory } The Catholic University of America } Hannan Hall ñ Rm 433 } Washington, D.C. 20064 } (202) 319-6237 (Office) } (202) 319-5346 (Lab) } (202) 319-4469 (Fax) } } } --------------------------------------------------------------------------- } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 09:06:37 2004
Cavin Mooers wrote: } } Question: We are currently experiencing vacuum } issues which are causing tungsten oxide growth } and premature failure of the filament while } operating our JEOL 5900. One of the JEOL } technicians has suggested that 5x10-4 torr is } acceptable, but I find this hard to believe...
We have a JEOL JXA-8900 electron microprobe, and we never expose a hot filament to pressures above 1x10-4 torr -- and that is only for a minute or so during a sample change. When, on occasion, a user has exceeded this pressure for a few seconds, our filament tends to break within a day or two. I would consider a pressure of 1x10-5 torr to be minimally acceptable, and we normally operate at about 2x10-6 torr. Once or twice we've had vacuum problems, and it has been the fault of degraded rubber gaskets causing leaks.
Ellery
--------------- Ellery E. Frahm Research Scientist/Manager Electron Microprobe Laboratory Department of Geology & Geophysics University of Minnesota - Twin Cities Lab Website: http://probelab.geo.umn.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 11:05:01 2004
I used to use a home-made glow-discharge device for grids and a lot more besides. I can't find the paper so I'm afraid I can't give you the reference, but I can tell you that the device is cheap and easy to make if you have access to a workshop, and it does work very well. That's assuming that you have the same paper that we used, of course. I never tried keeping grids in the fridge, but before we had the GD thing I used to use an anti-static pen from Ladd (I think - one of the usual suppliers, anyway) and that certainly helped. Sorry to be so vague, but I'm at home.
Lesley Weston.
on 14/01/2004 3:06 PM, Jon Krupp at 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 } ---------------------------------------------------------------------------- --} - } } Greetings: } } What's the latest word on glow discharge for grids? } } We have an old VE that will glow discharge, but have hardly ever used it. } A new faculty person wants to do more glow discharging and is looking for } info. } } Somewhere I have a paper describing a home made glow discharge device, } anybody know about this and if it works? } } Also have heard that keeping grids in the refrigerator helps too. What's up } with that? } } Her application is carbon films for negatively stained macromolecules. } } Thanks } } 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 Thu Jan 15 11:30:39 2004
Hi Shannan I have followed this theme with interest as we have used a Pelco Microwave for about four years now. Protocols and equipment have changed over that time and we have kept up with the changes. They have certainly improved the timing it takes to either fluoresenctly label cells for confocal (half an hour instead of 3 hours), or for EM processing (two hours instead of three days). If it works conventionally it is likely to work in the microwave. If it doesn't work conventionally then it is unlikely to work in the microwave. The results are certainly comparable and for confocal work, the cells are fresher and there is generally less background. Researchers can walk in the door at 9 am and go down to the confocal at 9.30 with labelled cells.
Our protocols have changed somewhat over this time and we are constantly finding better ways of doing things. Three things are important in your microwave: 1: a cool spot. This cuts out the standing waves and the need for load coolers and controls the temperature together with the temperature probe. 2. a vacuum chamber, especially for plant material or specimens with a cuticle such as C. elegans and beetle larvae. 3. Power controller - keep the power below 200 watts and live specimens stay alive.
The present protocol below has been used successfully with a wide variety of specimens (we are a multiuser facility for the faculty of medicine and the faculty of science). You may need to tweak the timings depending on how thick your samples are. You can easily check the penetration by sacrificing one of the larvae after the osmium step, though this is probably not a problem for SEM. Our TEM protocol starts the same way but we use a Spurr-Epon mix for the resin. The blocks cut better than using either Spurr's or Epon alone. You can of course use your buffer of choice. Cacodylate buffer is our choice as we get less chance of precipitates with UA and it is a lot cheaper than PIPES or HEPES though they have their places too.
Microwave SEM processing for animal tissue
1. Fixation Fix tissue in 2.5% Glutaraldhyde in 0.1M cacodylate buffer pH 7.3-7.4 at 22 C Perform under Vacuum Power level 1 (about 100W) 2 min on, 2 min off, 2 min on Repeat without changing
2. Cacodylate buffer rinse at 22 C Do not perform under Vacuum Power level 2 (about 200W) 40 sec Repeat three times If using tannic acid with buffer, rinse in butter once without tannic before continuing to osmium step.
3. Osmium Tetroxide Fixation at 22 C 1% osmium tetroxide in 0.1M cacodylate buffer Perform under Vacuum Power level 1 (about 100W) 2 min on, 2 min off, 2 min on Repeat without changing
4. Distilled water rinse Rinse sample in distilled water and change to new water Do not perform under Vacuum Power level 2 (about 200W) 40 sec
5. Dehydrate in ethanol at 22 C 50% ETOH 70% ETOH 90% or 95% ETOH 100% ETOH 100% ETOH 100% ETOH Do not perform under Vacuum Power level 2 (about 200W) 40 sec
6. Critical Point Dry step either conventionally in a CPD or in the microwave with Hexamethyldisilizane (HMDS) at 22 C
Do not perform under Vacuum Power level 2 (about 200W) 40 sec Replace with new HMDS Repeat two more times
Place in 60C oven for 5 mins Remove excess HMDS Place back in 60C oven until HMDS has evaporated
No two specimens types are the same. You might have to double the timings for the larvae depending on how difficult it is to penetrate the cuticle and how big they are.
A great souce of what is new in the processing of specimens is Rick Giberson of Pelco. He is working with a number of labs on improving the technique. I have no doubt that the above is already out of date, but it usually works for us, so if it ain't broke, don't fix it eh!
We have been investigating the use of formaldehyde in the microwave. I had a coop student look at the effect of the microwave on fixation over 30 mins, 2 hours and 72 hours. The first results have given us more questions. Hopefully we will be able to present a paper on this in the future.
Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 11:34:21 2004
HI. I would like to know how to measure the size of lattice of very local places in the HRTEM image (not real lattice dimensions, just the size between any lattice planes). The images are two two dimensional (nearly structural images), and the structure is relative simple, composed of two or three different atoms. Probably slightly tilting affects the measurement, but I want to know the size of each one or several lattice layers from the interface. I also want to know the lattice size of very local places (~10 x ~10 lattices). I think that some programs may find the lowest or highest contrast (1 pixel) on digital images, although I am not sure if the idea is correct. Please advise about some papers or programs. I prefer a free software.
Thank you,
Hiromi Konishi The University of New Mexico
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 11:53:57 2004
} ... we never expose a hot filament to } pressures above 1x10-4 torr -- and that is } only for a minute or so during a sample change.
Michael O'Keefe wrote:
} Never = "only for a minute or so"........ } Is that a new definition of "never"? } } Mike
Hi Mike,
Let me clarify:
We never deliberately expose a hot filament to pressures above 1x10-4 torr. We normally operate at about 2x10-6 torr. Only during a sample change will we reach pressures that *approach* 1x10-4 torr for a minute or so, often less. I tell our users never to exceed 1x10-4 torr, but sometimes a user is careless or distracted and will reach higher pressures for a few seconds, greatly reducing our filament life -- it usually "dies" in a day or two.
Sorry for the confusion -- I wasn't trying to re-define "never" :)
Ellery
--------------- Ellery E. Frahm Research Scientist/Manager Electron Microprobe Laboratory Department of Geology & Geophysics University of Minnesota - Twin Cities Lab Website: http://probelab.geo.umn.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 11:57:40 2004
The paper describing a home made glow-discharge device is authored by Aebi and Pollard, J Electron Microsc Tech. 1987 Sep; 7(1): 29-33. We made a similar one (a bit simpler) which we routinely use to charge grids prior to picking up sections and also prior to negative staining. It can probably be made for less than $150.00. I'd be happy to send a .jpg image of the device to anyone who wants it.
Doug
On Wed, 14 Jan 2004 15:06:29 -0800 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 } ------------------------------------------------------------------------------- } } Greetings: } } What's the latest word on glow discharge for grids? } } We have an old VE that will glow discharge, but have hardly } ever used it. A new faculty person wants to do more glow } discharging and is looking for info. } } Somewhere I have a paper describing a home made glow } discharge device, anybody know about this and if it works? } } Also have heard that keeping grids in the refrigerator } helps too. What's up with that? } } Her application is carbon films for negatively stained } macromolecules. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
---------------------- Douglas R. Keene Associate Investigator Micro-Imaging Center Shriners Hospital for Children 3101 S.W. Sam Jackson Park Road Portland, Oregon 97239-3009 phone: 503-221-3434 FAX: 503-412-6894
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 12:23:05 2004
On Jan 15, 2004, at 9:39 AM, Hiromi Konishi wrote:
} I would like to know how to measure the size of lattice of very local } places } in the HRTEM image (not real lattice dimensions, just the size between } any } lattice planes). } The images are two two dimensional (nearly structural images), and the } structure is relative simple, composed of two or three different atoms. } Probably slightly tilting affects the measurement, but I want to know } the } size of each one or several lattice layers from the interface. I also } want } to know the lattice size of very local places (~10 x ~10 lattices). I } think } that some programs may find the lowest or highest contrast (1 pixel) } on } digital images, although I am not sure if the idea is correct. Please } advise } about some papers or programs. I prefer a free software. } Dear Hiromi, I would first calibrate the microscope magnification using a Mag*I*Cal--no affiliation except satisfied user--then record the images on film, print enlargements, scan, and measure the areas of interest. This avoids at least some of the aliasing that can come from digital imaging. If you take digital images, you have to be sure that Nyquist frequency, equal to twice the pixel dimension, is larger than the spacing you are trying to determine; i.e., the lattice layers must span several pixels. If you know that all the layers have the same spacing, you can measure the spacing between layer 1 and layer N, but your post indicates that you are trying to measure the difference in spacing of layers near an interface--a much more difficult problem. 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 Thu Jan 15 14:33:28 2004
Our venerable JEOL 840A valves over to the diffusion pump about 3 minutes after we start pumping the chamber. 45 seconds later, the high voltage was re-enabled at a vacuum of 5x10-5 torr measured at the gun. At 30 minutes, vacuum had reached 1x10-5 torr. At 60 minutes, the vacuum was about 2x10-6. I think we get down into the high 10-7 range if we leave the scope overnight. (FYI, it takes less than 60 seconds for us to pump down just the gun chamber when we vent it and leave the sample chamber under vacuum. I don't know your model of JEOL. It may not have a valve to isolate the gun from the sample chamber.)
It definitely sounds like a leak or a pumping problem. I would defer to the other posters and their suggestions on those subjects.
You did not say what kind of samples you are examining, or whether samples are loaded when you are having trouble reaching vacuum. I know that can be a problem. We tried to examine concrete in our 840A and it took forever to pump down. We did a bit better by limiting sample volume, but it was still slow. Oily samples can be a problem depending on the vapor pressure of the liquid.
You were not clear whether your scope is under service contract or not. If it is, I would suggest you take advantage of the contract. Something is not right. If you can't find the problem quickly, I would let the boys earn their keep. I might assume this is a scope you purchased directly from JEOL, but that might not be the case. Did the scope used to work right in your lab and this is a new problem, or is this a matter of trying to get a scope up and running right for the first time?
Good luck and keep the questions coming. Warren
At 09:00 AM 1/15/2004, Richard Edelmann wrote:
} Calvin: } } Yes, 5x10-4 is a useable vacuum, but it will shorten fialment } life. As } previously suggested you may have a vacuum leak in the gun (check the gun } chamber o-ring). But since you are getting down to 5x10-6 it doesn't sound } like a leak is the main problem. Becasue it takes so long to pump down, } sounds more like a pumping problem. Not exactly sure where your starting } from when you say it takes 1 hour to get 3 x10-5. I just vented my JEOL 840 } gun and it took ~ 9 mins to get the same with a warmed and running pumping } system. } } -- dirty or cracked DP oil or RP oil (Change oils) } } -- poor DP backing (clean RP and replace RP exhaust filter). } } -- low RP oil level or DP oil level. } } -- check cooling temp on DP. (Too high *or* too low) } } } Good luck. } } } Email: cavinm-at-vsl.cua.edu } } Name: Cavin Mooers } } } } Organization: Vitreous State Lab } } } } Title-Subject: [Microscopy] [Filtered] SEM: Minimum Vacuum for } Tungsten Filament } } } } Question: We are currently experiencing vacuum } } issues which are causing tungsten oxide growth } } and premature failure of the filament while } } operating our JEOL 5900. One of the JEOL } } technicians has suggested that 5x10-4 torr is } } acceptable, but I find this hard to believe. The } } issue we are dealing with is extraordinarily slow } } pumping times -- 1 hour to obtain 3x10-5 torr and } } a max vacuum of 5x10-6 torr after 3-4 hours. We } } are a high volume lab, and so I wish to know what } } the minimum vacuum I need without seriously } } diminishing filament life, in order to maximize } } the workload. } } } } Sincerely, } } } } Cavin T. F. Mooers } } --------------------------------------------------------------------------- } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 350 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu
------------------------------------------- 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
Does anybody know of a current supplier of (preferably small lots) of Sylguard (Dow-Corning)? I've been searching the web, but don't find any suppliers. Sylguard as used for potting electronics, the clear stuff (although clear isn't required right now). A mold-release compound for this would be nice, too. Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 17:00:42 2004
Hiromi, If you have access to a Macintosh Version of Gatan's DigitalMicrograph, you can download from the NCEM web site a powerful plug-in written by Roar Kilaas and Martin Hytch. Using this plug-in, you can measure the local change in lattice parameter from an image. The program does a lot more and is very easy to use. See the link below:
http://ncem.lbl.gov/frames/software.htm ********************* A new set of routines for creating digital Moire patterns, displacement maps and strain images can be downloaded by clicking on the link below. These routines use the concept of the geometric phase to calculate deviations in local lattice parameters from variations in reciprocal space around chosen spatial frequencies (Bragg reflections). On-line help on the routines is available from the menu-bar.
Download Phase-Extension routines.
PhaseManual.pdf
Last updated May 1999.
Users of this package are encouraged to email comments (email: roar-at-lbl.gov) on the software. ********************
Cheers, Ray
Ray D. Twesten, PhD. Center for Microanalysis of Materials Seitz Materials Research Laboratory 104 S. Goodwin Ave., Urbana, IL 61801 +1 217 244-6177 (Fax -2278)
-----Original Message----- } From: Hiromi Konishi [mailto:konishi-at-geofourpeaks.com] Sent: Thursday, January 15, 2004 11:40 AM To: microscopy-at-ns.microscopy.com
HI. I would like to know how to measure the size of lattice of very local places in the HRTEM image (not real lattice dimensions, just the size between any lattice planes). The images are two two dimensional (nearly structural images), and the structure is relative simple, composed of two or three different atoms. Probably slightly tilting affects the measurement, but I want to know the size of each one or several lattice layers from the interface. I also want to know the lattice size of very local places (~10 x ~10 lattices). I think that some programs may find the lowest or highest contrast (1 pixel) on digital images, although I am not sure if the idea is correct. Please advise about some papers or programs. I prefer a free software.
Thank you,
Hiromi Konishi The University of New Mexico
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 17:30:30 2004
Jonathan Krupp wrote: ==================================================== What's the latest word on glow discharge for grids?
We have an old VE that will glow discharge, but have hardly ever used it. A new faculty person wants to do more glow discharging and is looking for info.
Somewhere I have a paper describing a home made glow discharge device, anybody know about this and if it works?
Also have heard that keeping grids in the refrigerator helps too. What's up with that?
Her application is carbon films for negatively stained macromolecules. ======================================================= Usually the reason why one would want to glow discharge treat their (carbon coated) grids is to make them more hydrophilic. The technique we use is to expose the carbon coated grids to a nitrogen (e.g. air) plasma for roughly 5-10 seconds in one of our standard configured Plasma Prep II plasma etcher units. Such a treatment will keep the carbon coatings hydrophilic for roughly 30 days (or more). More information can be found on URL http://www.2spi.com/catalog/instruments/etchers1.shtml
This is a low power unit and I don't know how well it might work for systems putting out more than 100 watts. I have never heard of storage under refrigeration to slow down the loss in hydrophilic character of carbon coated grids.
Disclaimer: SPI Supplies manufactures carbon coated grids for customers and also manufactures the SPI Plasma Prep™ II plasma etcher.
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 Jan 15 19:23:22 2004
A medium 10-6 torr in one hour sounds reasonable for diffusion pump powered scope. There are two things to consider: - it may be a leak (perhaps some O-ring); - it may be pumps (backing or diffusion).
If there is a leak, you need to isolate somehow (depends from the model) the suspected compartment and measure the leak speed - JEOL guys would tell you what is the normal for your particular microscope.
The pumps tend to lost their "power" with time if you don't do a prophylactic. The oil ether in mechanical or diffusion pump should be changed from time to time. It's more critical for mechanical pump, because it has moving parts. It's good idea to change oil in mechanical pump at least ones in a year. You also should keep in mind the possibility of backstreaming from mechanical pump, if so, diffusion pump may be affected. If diffusion pump operates in normal condition (not exposed to air when hot, not much water pumped down etc) the oil may stay good for years. Still, you need to maintain the level of oil according to the specification. If you did not manage to look inside of DP for decade, it's probably time to do so. Look for dark deposits and oil's color. If your DP has been operated in good conditions, you, perhaps will not find any dark deposits and oil will be from yellow to light dark (you lucky guy). So, because you already opened DP, you may need to replace oil for the next few years. If your DP is contaminated by deposits and oil is dark - it's time to do good cleaning. Disassemble everything and clean. Everything inside the DP should be shiny polished -the warranty, it will thankfully works good for the next decade! Return everything back and check leak speed again. Personally, I prefer to use Santovac-5 DP oil. It's expensive but it delivers wonderful results and it's very stable. In my DV502A vacuum evaporator, it stays for 10 years and I only adjust level from time to time. It delivers good 10-6 in about 1-2 hours and goes into 10-7 overnight. It's on the "dirty" system. Sergey
At 12:38 PM 1/15/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
What's the latest word on glow discharge for grids?
Another way to make coated grids more hydrophilic is to expose them to UV light. I know of one lab that stored Formvar-coated grids on racks under a UV light (did not specify wavelength), and used the oldest ones first. I personally just make grids on a dry day and let them age naturally - probably enough UV here to do the job.
But more specifically, a student here at UH tried a lot of different methods of treating her grids, and found that if she put them in their Stratolinker UV Crosslinker (for crosslinking DNA), set it for 30 sec, and pushed the Auto button, it worked great! I looked it up - it uses 254 nm. I have not yet tried any of our UV sources around here, but it would be worth trying everything from a party blacklight to a zap with the confocal. Or even a turn on the front porch (weather permitting).
Aloha, Tina
Yesterday - rainy, gale force winds Today - 78F, sunny, surf's up
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 15 23:50:24 2004
Email: gt5185d-at-prism.gatech.edu Name: Jonathan B.
Organization: Georgia Tech
Title-Subject: [Microscopy] [Filtered] MListserver: SEM analysis of skin tissue...
Question: Hello Microscopy group,
I was hoping to garnish some advice about SEM sample preparation for my PhD research. I'm investigating a technology to microporate human skin for transdermal drug delivery, and I want to image the pores with a SEM. My current plan is to purchase an automated critical point dryer; currently I'm considering Emitech's 850 and Polaron's 7501. I hope I'm on the right track.
Any comments? Thanks, Jonathan B.
My biological samples are engineered living tissue 22mm in diameter and 1mm thick. I'll be using a Hitachi 3500H, with a CVC Products DC Sputterer at the Georgia Tech MiRC cleanroom.
I don't bother with neutralization or anything; I just collect it in a jug and hand it over to Rutgers Environmental Health & Safety when they come to pick up hazardous waste.
Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology and Toxicology Ernest Mario School of Pharmacy Rutgers University
Barbara Murray wrote:
} Greetings, } We have been using a solidifier for our formalin, putting it into biohazard boxes for pickup by the housekeeping dept. We were told by our Safety Officer that we can now pour it down the drain with lots of running water. } For the ones of you who are using formalin, how are you disposing of it? } Thanks for your replies. Have a great day and weekend! } } Barbara A. Murray,HT.(ASCP) } The Alaska Native Medical Center } Anchorage, Alaska } } } _______________________________________________ } Histonet mailing list } Histonet-at-lists.utsouthwestern.edu } http://lists.utsouthwestern.edu/mailman/listinfo/histonet } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 16 09:00:59 2004
Picking up on the thread on disposal of formalin, I have a comment plus a question. My comment is that many Environmental Health & Safety departments pick up "hazardous waste" and then pour it down the drain with lots of water. This includes low level radioisotopes. At my university, we (lab personnel) can not call anything hazardous waste. We can only have "unwanted used materials". We can not use the word waste on any label. We had a box labeled "glass waste" on our microtome bench for our old glass knives and slides and were cited!
Recently we were told they were going to a policy of users disposing uranyl acetate ( {1%) by pouring it down the drain with lots of water. I was a little surprised by this. Do other universities follow this policy?
Thanks, Tom
At 09:33 AM 1/16/2004 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
We are not allowed to throw anything down the sink--including water wash from stains and alcohol! The MWRA (Massachusetts Water Resource Assoc.) has very strict guidelines re: mercury, etc. entering Boston Harbor. As we are a pathology lab for a research group, we do a lot of staining, so all washes must be collected in a large container and it is then picked up by an outside waste management company on a weekely basis. This waste is analyzed for mercury levels. You would be surprised the number of chemicals that contain mercury. All our other waste (i.e. formalin, EM fixes, etc.) are collected in containers and picked up weekly as well. I'm surprised that, being in Alaska, where there have been problems with major spills in the waters, that they allow formalin to go down the drain. Are you allowed to throw other chemicals down as well?
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] Sent: Friday, January 16, 2004 9:34 AM To: Barbara Murray; Microscopy-at-sparc5.microscopy.com
I don't bother with neutralization or anything; I just collect it in a jug and hand it over to Rutgers Environmental Health & Safety when they come to pick up hazardous waste.
Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology and Toxicology Ernest Mario School of Pharmacy Rutgers University
Barbara Murray wrote:
} Greetings, } We have been using a solidifier for our formalin, putting it into biohazard boxes for pickup by the housekeeping dept. We were told by our Safety Officer that we can now pour it down the drain with lots of running water. } For the ones of you who are using formalin, how are you disposing of it? } Thanks for your replies. Have a great day and weekend! } } Barbara A. Murray,HT.(ASCP) } The Alaska Native Medical Center } Anchorage, Alaska } } } _______________________________________________ } Histonet mailing list } Histonet-at-lists.utsouthwestern.edu } http://lists.utsouthwestern.edu/mailman/listinfo/histonet } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 16 10:27:19 2004
I refer you to my general email that I just sent to the group--we cannot throw anything down the sink. But, especially, uranly acetate, which has a low level of radioactivity. This is picked up by the radiation safety dept. I believe they are just storing it until decisions are made as to what waste site it can be shipped.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Friday, January 16, 2004 10:06 AM To: Microscopy-at-msa.microscopy.com
Picking up on the thread on disposal of formalin, I have a comment plus a question. My comment is that many Environmental Health & Safety departments pick up "hazardous waste" and then pour it down the drain with lots of water. This includes low level radioisotopes. At my university, we (lab personnel) can not call anything hazardous waste. We can only have "unwanted used materials". We can not use the word waste on any label. We had a box labeled "glass waste" on our microtome bench for our old glass knives and slides and were cited!
Recently we were told they were going to a policy of users disposing uranyl acetate ( {1%) by pouring it down the drain with lots of water. I was a little surprised by this. Do other universities follow this policy?
Thanks, Tom
At 09:33 AM 1/16/2004 -0500, you wrote:
} --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
We would like to prepare sperm for examination under EM of cross sections of the tail to investigate integrity of the cilia, and I have never done this before. Does anyone have a good simple method that would give a good result?
Garry Burgess Charge Technologist Health Sciences Centre Winnipeg, Canada
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 16 12:22:01 2004
Gary, I just finished doing a sperm prep. I had the investigator pellet them (very gently to minimize head-tail separation) and fix the resultant pellet in my "standard" EM fix (2.5% glut, 4% pfa, 0.02% picric acid [good for membranes] in 0.1M Na-cacodylate). I processed the pellet as usual with an osmium postifx, ethanol dehydrations and i embedded in Spurr's. Very straight forward and we got some lovely images. You do need to hunt around for good cross sections, but there are usually so many sperm in the pellet its not too hard. 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 Jan 16 12:59:24 2004
} We would like to prepare sperm for examination under EM of cross sections of } the tail to investigate integrity of the cilia, and I have never done this } before. Does anyone have a good simple method that would give a good } result? } } Garry Burgess } Charge Technologist } Health Sciences Centre } Winnipeg, Canada Garry, I have worked with marine animal sperm in the past. An important piece of information was that we used artificial sea water as the carrier for the glutaraldehyde instead of a buffer and as the wash after that. The best pelleted samples were obtained if the tubes were centrifuged as soon as possible after the primary fixative was added to the tube. If the pellet falls apart, centrifuge during each change of solution. At the time, the top speed of a table top centrifuge was used but a microfuge should work better. Care should be taken that the pellet is not thick. Acetone (Mallinckrodt #2440) was chosen over ethanol because we were interested in the cytoskeletin and membranes, and wanted to remove a lot of background substances.
Procedure: 1% glutaraldehyde in sea water [1 part 8% glut from Electron Microscopy Sciences + 7 parts sea water], room temp. 30 min. sea water rinse 1% osmium tetroxide in 0.1 M phosphate buffer, on ice + dark, 30 min. Cold Water rinse X3, 5 min. each 1% UA in water, refrigerator, overnight Acetone dehydration - 50% to 100% X2, 15 min. each Propylene Oxide X2, 15 min. each 1:1::Prop.Ox.:Epon 30 min. Epon 30 min. Embed in Epon and polymerize.
All times may be extended except the osmium fixation, especially if actin is of importance.
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 16 13:27:34 2004
} Any idea how long the effect lasts? For us it seemed like the effect did } not last very long at all. But then again, maybe we did not study the } phenomenon to the extent you did.
A very good question. I know they are using their grids immediately. The people who kept their grids on racks under a UV light left the light on all the time, and used the grids soon after taking them out.
Another client of ours has used bacitracin and protein A (not together) to help his viruses stick and to increase wettability of Formvar coated grids with great success. He is no longer here, so I don't have his protocols.
When in desparation (a chronic state around here) I have tried a number of techniques for making coated grids more hydrophilic. The more successful ones include dipping a grid in 70-80% ethanol, shaking off the excess, and then using the grid just as the fluid appears to dry, and I have used a very dilute solution of PhotoFlo, which worked surprisingly well for the application at hand and did not leave a visible residue. I have not yet been desparate enough to try spit!
Caroline Schooley, when at Berkeley, used to have a homemade (I think) Tesla coil kind of thing that she applied to the outside (I think) of the bell jar of a vacuum evaporator. I don't remember well because I was terrified of the thing and usually ran out of the room. I was very young.
In general, however, I coat a lot of grids on our rare dry day, then keep them in covered Petri dishes. For Formvar-coated grids, I like them best at about 2 years old, and for carbon films at 6 months or more. I don't know why the become more hydrophilic as they age, and I'm guessing it's some kind of contamination, but I haven't seen anything weird, and they work well for me.
This is all to keep from having to repair my vacuum evaporator, of course, but glow discharge is probably the most reliable. My new sputter coater works, however. Chuck, how about plasma mode? What does that do?
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 16 14:56:43 2004
Nope, uranyl acetate is treated as radioactive waste here. I don't know what they do with it after it leaves here, however.
Kathleen Roberts Rutgers University
Tom Phillips 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 } ------------------------------------------------------------------------------- } } } Picking up on the thread on disposal of formalin, I have a comment } plus a question. My comment is that many Environmental Health & } Safety departments pick up "hazardous waste" and then pour it down the } drain with lots of water. This includes low level radioisotopes. At } my university, we (lab personnel) can not call anything hazardous } waste. We can only have "unwanted used materials". We can not use } the word waste on any label. We had a box labeled "glass waste" on } our microtome bench for our old glass knives and slides and were cited! } } Recently we were told they were going to a policy of users disposing } uranyl acetate ( {1%) by pouring it down the drain with lots of water. } I was a little surprised by this. Do other universities follow this } policy? } } Thanks, Tom } } } } } At 09:33 AM 1/16/2004 -0500, you 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 don't bother with neutralization or anything; I just collect it in } } a jug and hand it over to Rutgers Environmental Health & Safety when } } they come to pick up hazardous waste. } } } } Kathleen Roberts } } Principal Lab Technician } } Neurotoxicology Labs } } Dept of Pharmacology and Toxicology } } Ernest Mario School of Pharmacy } } Rutgers University } } } } Barbara Murray wrote: } } } } } Greetings, } } } We have been using a solidifier for our formalin, putting it into } } } biohazard boxes for pickup by the housekeeping dept. We were told } } } by our Safety Officer that we can now pour it down the drain with } } } lots of running water. For the ones of you who are using formalin, } } } how are you disposing of it? } } } Thanks for your replies. Have a great day and weekend! } } } } } } Barbara A. Murray,HT.(ASCP) } } } The Alaska Native Medical Center } } } Anchorage, Alaska } } } } } } } } } _______________________________________________ } } } Histonet mailing list } } } Histonet-at-lists.utsouthwestern.edu } } } http://lists.utsouthwestern.edu/mailman/listinfo/histonet } } } } } } } } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 16 16:01:28 2004
Tina Carvalho wrote about glow discharge: } } Caroline Schooley, when at Berkeley, used to have a homemade (I } think) Tesla coil kind of thing that she applied to the outside (I } think) of the bell jar of a vacuum evaporator. I don't remember well } because I was terrified of the thing and usually ran out of the room. I } was very young.
Before someone yells at me, let me correct Tina's memory. I used a physics demonstration-type Tesla coil in firm contact (to avoid ozone production in the room) with a current feedthrough that led from below the baseplate into the bell jar. Ran the discharge during the rough pump part of the automatic pumpdown cycle, about a minute, until the purple glow inside the bell jar faded. Worked fine; I used it for years.
Young and terrified? By then you could completely rebuild a Volkswagen, Tina....! -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 16 16:33:57 2004
The regulations here have changed many times for UA disposal. It went to sink, then to Chemical waste, then it was too hot for them, so it was to go to Radiation safety, but it was not hot enough... Right now I have given up and have an old, covered, tri-pour beaker in the corner of my hood with a combination of evaporated UA and uranyl phosphate (UA+PO4 Buffer)! Maybe I'll have them put it into my coffin when my time comes!
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145 psconnel-at-sas.upenn.edu
} Nope, uranyl acetate is treated as radioactive waste here. I don't } know what they do with it after it leaves here, however. } } Kathleen Roberts } Rutgers University } } } Picking up on the thread on disposal of formalin, I have a comment } } plus a question. My comment is that many Environmental Health & } } Safety departments pick up "hazardous waste" and then pour it down } } the drain with lots of water. This includes low level } } radioisotopes. At my university, we (lab personnel) can not call } } anything hazardous waste. We can only have "unwanted used } } materials". We can not use the word waste on any label. We had a } } box labeled "glass waste" on our microtome bench for our old glass } } knives and slides and were cited! } } } } Recently we were told they were going to a policy of users } } disposing uranyl acetate ( {1%) by pouring it down the drain with } } lots of water. I was a little surprised by this. Do other } } universities follow this policy? } } } } Thomas E. Phillips, PhD } } Professor of Biological Sciences } } Director, Molecular Cytology Core } } 3 Tucker Hall } } University of Missouri } } Columbia, MO 65211-7400 } } 573-882-4712 (office) } } 573-882-0123 (fax) PhillipsT-at-missouri.edu
} } } I don't bother with neutralization or anything; I just collect it } } } in a jug and hand it over to Rutgers Environmental Health & Safety } } } when they come to pick up hazardous waste. } } } } } } Kathleen Roberts } } } Principal Lab Technician } } } Neurotoxicology Labs } } } Dept of Pharmacology and Toxicology } } } Ernest Mario School of Pharmacy } } } Rutgers University } } } } } } Barbara Murray wrote: } } } } } } } Greetings, } } } } We have been using a solidifier for our formalin, putting it into } } } } biohazard boxes for pickup by the housekeeping dept. We were } } } } told by our Safety Officer that we can now pour it down the drain } } } } with lots of running water. For the ones of you who are using } } } } formalin, how are you disposing of it? } } } } Thanks for your replies. Have a great day and weekend! } } } } } } } } Barbara A. Murray,HT.(ASCP) } } } } The Alaska Native Medical Center } } } } Anchorage, Alaska } } } } _______________________________________________ } } } } Histonet mailing list } } } } Histonet-at-lists.utsouthwestern.edu } } http://lists.utsouthwestern.edu/mailman/listinfo/histonet
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 16 18:31:28 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (echeung-at-eyetk.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, January 16, 2004 at 13:40:06 ---------------------------------------------------------------------------
Question: Has anyone had success with using nanogold labeling in whole-mount preparations? What did you use to improve contrast of cellular structure without using osium tetraoxide?
} Recently we were told they were going to a policy of users disposing } uranyl acetate ( {1%) by pouring it down the drain with lots of water. } I was a little surprised by this. Do other universities follow this } policy? } Dear Tom, There are different laws in different states, so where you live determines what is possible. Within those limits, your safety office may impose more stringent conditions. Disposal of low-level radioactive waste--oops, spent materials--is expensive, so many institutions allow only the least costly option. 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 Sat Jan 17 19:01:42 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gbarclay-at-fans.uwi.tt) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, January 17, 2004 at 11:26:01 ---------------------------------------------------------------------------
Email: gbarclay-at-fans.uwi.tt Name: Greg Barclay
Organization: University of the West Indies
Education: Graduate College
Location: St. Augustine, Trinidad, West Indies
Question: Permount is what we have been using forever and we are now looking for a slide mountant that dries faster. I would appreciate any suggestions.
I am considering trying to budget for an Osmium-based specimen coater to augment (not replace) my Denton Desk II Au/Pd and Pt coater. There are some specimens that are going to be taken at high mag (250KX-450KX) and should not show coating artifacts (major or minor). The Os coater looks like a good option. Cr is out due to short life span based on rapid oxidation. Is the same true for Os?
I would appreciate feedback from Os coater users and suppliers. Off-list of course.
tnx, gary g.
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 18 23:03:46 2004
Gary Gaugler wrote: ======================================================= I am considering trying to budget for an Osmium-based specimen coater to augment (not replace) my Denton Desk II Au/Pd and Pt coater. There are some specimens that are going to be taken at high mag (250KX-450KX) and should not show coating artifacts (major or minor). The Os coater looks like a good option. Cr is out due to short life span based on rapid oxidation. Is the same true for Os?
I would appreciate feedback from Os coater users and suppliers. Off-list of course. ========================================================= Osmium is a precious group metal and therefore it has essentially the intertness of gold, platinum, etc. It is a pretty permanent coating, and could be expected to have a life time comparable to what one would expect for gold.
We are often times asked if there is any danger that it could convert to the tetroxide (and then sublime and disappear). From a practical standpoint, absolutely not. Of course, if you exposed the coating to a strong oxidizer, perhaps sodium iodide, the metallic osmium could be oxidized back up to the dioxide and then the tetroxide and then there would be a dangerous condition but most of us don't expose our coated SEM samples to strong oxidizers......
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 18 23:26:04 2004
Gary Gaugler wrote: ======================================================= I am considering trying to budget for an Osmium-based specimen coater to augment (not replace) my Denton Desk II Au/Pd and Pt coater. There are some specimens that are going to be taken at high mag (250KX-450KX) and should not show coating artifacts (major or minor). The Os coater looks like a good option. Cr is out due to short life span based on rapid oxidation. Is the same true for Os?
I would appreciate feedback from Os coater users and suppliers. Off-list of course. ========================================================= Osmium is a precious group metal and therefore it has essentially the intertness of gold, platinum, etc. It is a pretty permanent coating, and could be expected to have a life time comparable to what one would expect for gold.
We are often times asked if there is any danger that it could convert to the tetroxide (and then sublime and disappear). From a practical standpoint, absolutely not. Of course, if you exposed the coating to a strong oxidizer, perhaps sodium iodide, the metallic osmium could be oxidized back up to the dioxide and then the tetroxide and then there would be a dangerous condition but most of us don't expose our coated SEM samples to strong oxidizers!
Chuck
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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 Sun Jan 18 23:54:39 2004
Hello all, We are currently working with a student who is interested in corals and possible virus associations within them. Fo course, problems arise when trying to process and section the samples, which contain both normal soft biological tissue and the hard calcified material. Could anyone please suggest a method to decalcify them without doing too much damage to the ultrastructure? Should a decalcification step be done on fresh or fixed tissue? The samples we have to work with now are fixed. Any suggestions would be appreciated. Thanks.
Lyn Waterhouse Adelaide Microscopy University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 or 8303 5855 Fax: (08) 8303 4356 http://www.adelaide.edu.au/microscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 01:22:17 2004
The best decalcifier to use on biological samples is actually a chelating agent; EDTA ethylene-diaminetetracetic acid. It does not act like a normal acid but binds metallic ions, especially calcium and magnesium. It works better at at pH 7-8 and can be used as an aqueous solution or mixed with formaldehyde.It takes longer than the usual decalcifiers such as acids but the results are very good. Dense cortical bone takes about 6-8 weeks to decalcify. If you have x-ray facilities you can monitor the process well. Decalcification must be done on well fixed material otherwise the decalcifier will macerate the biological matter, particularly the nucleic acids.
Regards, Evelyn Kaplan, Dept of Pathology, College of Medicine and Health Sciences, Sultan Qaboos University, Oman
-----Original Message----- } From: Lyn Waterhouse [mailto:lyn.waterhouse-at-adelaide.edu.au] Sent: Monday, January 19, 2004 10:03 AM To: Microscopy-at-MSA.Microscopy.Com
Hello all, We are currently working with a student who is interested in corals and possible virus associations within them. Fo course, problems arise when trying to process and section the samples, which contain both normal soft biological tissue and the hard calcified material. Could anyone please suggest a method to decalcify them without doing too much damage to the ultrastructure? Should a decalcification step be done on fresh or fixed tissue? The samples we have to work with now are fixed. Any suggestions would be appreciated. Thanks.
Lyn Waterhouse Adelaide Microscopy University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 or 8303 5855 Fax: (08) 8303 4356 http://www.adelaide.edu.au/microscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 02:21:36 2004
Marc Moeremans, who recently joined us, was one of the scientists involved in the development of the Nanovid microscopy technique at Janssen Pharmaceutics (Beerse, Belgium), which involved immunogold staining.
I forward your question to him, maybe he can help ?
=========================================== by way of Nestor J. Zaluzec 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 (echeung-at-eyetk.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Friday, January 16, 2004 at 13:40:06 } --------------------------------------------------------------------------- } } Email: echeung-at-eyetk.com } Name: Eunice Cheung } } Organization: Eyetech } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Has anyone had success with using nanogold labeling in } whole-mount preparations? What did you use to improve contrast of } cellular structure without using osium tetraoxide? } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 05:15:26 2004
Wonderful results on pre-embedding labeling vibratome sections of brain tissue have been described by Yi et al., (2001) J. Histochem. Cytochem. 49(3), 279-283. F(ab')2 and single Fab fragments coupled to ultra-small gold particles were used to immunolabel different intra cellular antigens. The detection of MGP-160, a golgi marker localized within the lumen demonstrates the potential of these ultra-small gold conjugates. Hong Yi uses osmium tetroxide to reveal morphological detail. She uses osmium after silver enhancing the ultra-small gold particles.
Similar protocols are used for whole mount preparations. A few examples: Briane et al; (2002) J. Histochem. Cytochem. 50(7), 983-991 Verbeek et al; (2002) J. Histochem. Cytochem. 50(5), 681-690.
More pre-embedding labeling protocols you can find in Aurion newsletter nr.5 Please contact me in case you need additional technical information.
Kind regards,
Peter
On Saturday, January 17, 2004, at 01:36 AM, by way of Nestor J. Zaluzec 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 (echeung-at-eyetk.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Friday, January 16, 2004 at 13:40:06 } ----------------------------------------------------------------------- } ---- } } Email: echeung-at-eyetk.com } Name: Eunice Cheung } } Organization: Eyetech } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Has anyone had success with using nanogold labeling in } whole-mount preparations? What did you use to improve contrast of } cellular structure without using osium tetraoxide? } } ----------------------------------------------------------------------- } ---- } } } ----------- Peter van de Plas Aurion Costerweg 5 6702 AA Wageningen The Netherlands Phone: +31 317 497676 Fax: +31 317 415955
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 07:01:46 2004
I have been given two samples and asked to determine if they are concrete, sandstone, or some mix. One sample is light gray, and I suspect may be mortar rather than concrete. The other sample has a slight rust/red hue and contains appreciably more calcium. Both samples contain silica, and both test positive for calcium carbonate by the acid drop test, with the light gray sample showing a yellow residue from this test.
Hours of reading test procedures for Portland cement and concrete, coupled with internet searches have left me wondering if I can draw a conclusion based on a few grams of sample. I would appreciate any suggestions about what to read or how to make this kind of determination.
Thanks,
Chris Holp FirstEnergy Corp. Beta Labs HolpC-at-firstenergycorp.com
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 07:27:39 2004
It seems to me..... Sandstone is an agglomeration of chiefly quartz in a matrix of carbonates and iron oxides. Concrete can have sand and small rocks or pebbles, but the assuming the concrete has reacted or set, it should be loaded with a variety of high refractive index (greater than 1.660 - which tells you where I got my training) particles. It the material is not cured or set up, you should be able to find high refractive index particles of calcium oxide which isn't found in sandstone.
Best wishes..............
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 08:05:00 2004
Chris, I'm not a geologist or a cement expert, but concrete and mortar have cement as part of their make-up (ingredient). So you probably should start out comparing sandstone and cement. Commonly, concrete is cement with crushed stones (aggregates) in it. You don't mention having EDS at your disposal. Having EDS spectra obtained on your sample, I would expect higher Si concentration in the sandstone. and higher Ca conc. in the cement. Drop/spots tests may not reveal conc. differences. You may also want to run standards (i. e. Portland Cement, etc.) with your samples for positive confimation. Or even call a cement company and find out how they analyze their products. Hope this helps.
The Materials Handbook, 13th ed. has some basic (starting) info on all three materials. ED
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
holpc-at-firstenergycorp.com 01/19/04 08:09 AM
To Microscopy-at-msa.microscopy.com cc
Subject SEM/LM of concrete vs. sandstone
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One and all,
I have been given two samples and asked to determine if they are concrete, sandstone, or some mix. One sample is light gray, and I suspect may be mortar rather than concrete. The other sample has a slight rust/red hue and contains appreciably more calcium. Both samples contain silica, and both test positive for calcium carbonate by the acid drop test, with the light gray sample showing a yellow residue from this test.
Hours of reading test procedures for Portland cement and concrete, coupled with internet searches have left me wondering if I can draw a conclusion based on a few grams of sample. I would appreciate any suggestions about what to read or how to make this kind of determination.
Thanks,
Chris Holp FirstEnergy Corp. Beta Labs HolpC-at-firstenergycorp.com
----------------------------------------- The information contained in this message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately, and delete the original message.
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 09:10:26 2004
You raise questions on some of the perplexing results from this job. While I am well aware of the heterogeneous nature of the samples, a summary of the "bulk" analysis results yielded (approx.): Lt. gray sample; O 38%, Si 40%, Ca 20%, with Mg, Al, and S also present. Rust/Red sample; O 48%, Si 4%, Ca 47%, with Mg, Al, S, and Fe also present.
A small piece from each sample was sanded through 320 grit paper to get a fresh, flat surface for the EDS analyses.
Now, neither sample is known to be concrete nor sandstone. Neither sample contains any aggregate material. The discrete particles present in the light gray sample are more populous and twice the size of the particles in the red material. There is no color banding in either sample, which could be indicative of the stratified layers in a sandstone.
I may be making this harder than it has to be, but I am suspecting that neither sample is sandstone.
ekomarnicki-at-MacDe rmid.com To: holpc-at-firstenergycorp.com cc: Microscopy-at-msa.microscopy.com 01/19/2004 09:13 Subject: [Microscopy] Re: SEM/LM of concrete vs. AM sandstone
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Chris, I'm not a geologist or a cement expert, but concrete and mortar have cement as part of their make-up (ingredient). So you probably should start out comparing sandstone and cement. Commonly, concrete is cement with crushed stones (aggregates) in it. You don't mention having EDS at your disposal. Having EDS spectra obtained on your sample, I would expect higher Si concentration in the sandstone. and higher Ca conc. in the cement. Drop/spots tests may not reveal conc. differences. You may also want to run standards (i. e. Portland Cement, etc.) with your samples for positive confimation. Or even call a cement company and find out how they analyze their products. Hope this helps.
The Materials Handbook, 13th ed. has some basic (starting) info on all three materials. ED
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One and all,
I have been given two samples and asked to determine if they are concrete, sandstone, or some mix. One sample is light gray, and I suspect may be mortar rather than concrete. The other sample has a slight rust/red hue and contains appreciably more calcium. Both samples contain silica, and both test positive for calcium carbonate by the acid drop test, with the light gray sample showing a yellow residue from this test.
Hours of reading test procedures for Portland cement and concrete, coupled with internet searches have left me wondering if I can draw a conclusion based on a few grams of sample. I would appreciate any suggestions about what to read or how to make this kind of determination.
Thanks,
Chris Holp FirstEnergy Corp. Beta Labs HolpC-at-firstenergycorp.com
----------------------------------------- The information contained in this message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately, and delete the original message.
----------------------------------------- The information contained in this message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately, and delete the original message.
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 09:33:16 2004
The difference between sandstones and concretes is usually fairly obvious in a polished thin section viewed under plain and crossed polars. The Atlas of sedimentary rocks under the microscope (Adams, MacKenzie and Guilford, 1984, Longmans) gives examples of the commonest types of sedimentary rock. A sandstone is composed of two components the clasts (sand/rock) and the cement (sorry that is the term the geologists use). Either of these components may contain calcium bearing minerals, so the simple presence of a signficiant calcium peak in an EDX spectra may not necessarily prove that the material is a concrete. However, the cement of concrete will not normally show visible crystallinity in the optical microscope whereas many sandstones will. It also allows you to identify the clasts and their size so distinguish between mortar and concrete (if samples is large enough to include representative clasts).
ekomarnicki-at-MacDermid.com 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 } ------------------------------------------------------------------------------- } } Chris, I'm not a geologist or a cement expert, but concrete and mortar } have cement as part of their make-up (ingredient). So you probably should } start out comparing sandstone and cement. Commonly, concrete is cement } with crushed stones (aggregates) in it. You don't mention having EDS at } your disposal. Having EDS spectra obtained on your sample, I would expect } higher Si concentration in the sandstone. and higher Ca conc. in the } cement. Drop/spots tests may not reveal conc. differences. You may also } want to run standards (i. e. Portland Cement, etc.) with your samples for } positive confimation. Or even call a cement company and find out how they } analyze their products. Hope this helps. } } The Materials Handbook, 13th ed. has some basic (starting) info on all } three materials. ED } } Ed Komarnicki } Sr. Analytical Chemist } MacDermid Inc. } } } } } } holpc-at-firstenergycorp.com } 01/19/04 08:09 AM } } To } Microscopy-at-msa.microscopy.com } cc } } Subject } SEM/LM of concrete vs. sandstone } } } } } } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------- } } One and all, } } I have been given two samples and asked to determine if they are } concrete, } sandstone, or some mix. One sample is light gray, and I suspect may be } mortar rather than concrete. The other sample has a slight rust/red hue } and } contains appreciably more calcium. Both samples contain silica, and both } test positive for calcium carbonate by the acid drop test, with the light } gray sample showing a yellow residue from this test. } } Hours of reading test procedures for Portland cement and concrete, coupled } with internet searches have left me wondering if I can draw a conclusion } based on a few grams of sample. I would appreciate any suggestions about } what to read or how to make this kind of determination. } } Thanks, } } Chris Holp } FirstEnergy Corp. } Beta Labs } HolpC-at-firstenergycorp.com } } } } ----------------------------------------- } The information contained in this message is intended only for the } personal and confidential use of the recipient(s) named above. If the } reader of this message is not the intended recipient or an agent } responsible for delivering it to the intended recipient, you are hereby } notified that you have received this document in error and that any } review, dissemination, distribution, or copying of this message is } strictly prohibited. If you have received this communication in error, } please notify us immediately, and delete the original message. } } } } } }
-- Chris Salter, Oxford Materials Characterisation Service, Oxford University Begbroke Science Park, Sandy Lane, Yarnton, Oxford, OX5 1PF Tel 01865 283722, EPMA 283741, Mobile 07776031608
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 10:05:44 2004
"Now, neither sample is known to be concrete nor sandstone. ..." Well that doesn't narrow it down any. According to my limited resource; cement would have } 3x more Ca than Si; and in sandstone the ratio of Si to Ca is expected to be much greater than double (what you have). I would guess that neither sample is sandstone.
Perhaps a geologist could guide you further. ED
holpc-at-firstenergycorp.com 01/19/04 10:18 AM
To ekomarnicki-at-MacDermid.com cc Microscopy-at-msa.microscopy.com Subject Re: Re: SEM/LM of concrete vs. sandstone
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You raise questions on some of the perplexing results from this job. While I am well aware of the heterogeneous nature of the samples, a summary of the "bulk" analysis results yielded (approx.): Lt. gray sample; O 38%, Si 40%, Ca 20%, with Mg, Al, and S also present. Rust/Red sample; O 48%, Si 4%, Ca 47%, with Mg, Al, S, and Fe also present.
A small piece from each sample was sanded through 320 grit paper to get a fresh, flat surface for the EDS analyses.
Now, neither sample is known to be concrete nor sandstone. Neither sample contains any aggregate material. The discrete particles present in the light gray sample are more populous and twice the size of the particles in the red material. There is no color banding in either sample, which could be indicative of the stratified layers in a sandstone.
I may be making this harder than it has to be, but I am suspecting that neither sample is sandstone.
ekomarnicki-at-MacDe rmid.com To: holpc-at-firstenergycorp.com cc: Microscopy-at-msa.microscopy.com 01/19/2004 09:13 Subject: [Microscopy] Re: SEM/LM of concrete vs. AM sandstone
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Chris, I'm not a geologist or a cement expert, but concrete and mortar have cement as part of their make-up (ingredient). So you probably should start out comparing sandstone and cement. Commonly, concrete is cement with crushed stones (aggregates) in it. You don't mention having EDS at your disposal. Having EDS spectra obtained on your sample, I would expect higher Si concentration in the sandstone. and higher Ca conc. in the cement. Drop/spots tests may not reveal conc. differences. You may also want to run standards (i. e. Portland Cement, etc.) with your samples for positive confimation. Or even call a cement company and find out how they analyze their products. Hope this helps.
The Materials Handbook, 13th ed. has some basic (starting) info on all three materials. ED
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
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One and all,
I have been given two samples and asked to determine if they are concrete, sandstone, or some mix. One sample is light gray, and I suspect may be mortar rather than concrete. The other sample has a slight rust/red hue and contains appreciably more calcium. Both samples contain silica, and both test positive for calcium carbonate by the acid drop test, with the light gray sample showing a yellow residue from this test.
Hours of reading test procedures for Portland cement and concrete, coupled with internet searches have left me wondering if I can draw a conclusion based on a few grams of sample. I would appreciate any suggestions about what to read or how to make this kind of determination.
Thanks,
Chris Holp FirstEnergy Corp. Beta Labs HolpC-at-firstenergycorp.com
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 12:21:16 2004
EDTA decalcification can be carried out on well-fixed specimens, including corals, in a laboratory (tempreature-controlled) microwave in a fraction of the time needed on the bench at room temperature. Stirring during the process is important to keep fresh decalcifying agent at the specimen at all times.
best regards, Steven Slap
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 15:23:41 2004
Two questions: 1. I was looking up the density of glycerol on the web and several sites gave it as 1.26 and several as 1.476. I had presumed the higher number but what is the other referring to? 2. In making a Mowiol mounting media as described in previous posts, I noted that the final glycerol concentration is only about 20% which is much lower than I am used to doing and seems a poor match in refractive index for oil. Does the Mowiol raise the refractive index to nearer the desired level? Thanks-Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 15:49:51 2004
JEOL vacuum systems are normally set to allow the filament to be turned on at 5X10-4 torr but, I personally don't recommend turning the gun on at this point, unless you are in a real hurry to get something done. Waiting 10-15 minutes after achieving a vacuum ready condition should get you in the high 10-5 torr range which would be more acceptable and extend filament life. Running at poor vacuum will also contaminate your column and apertures sooner which may contribute to your down time.
Taking several hours to get to 5X10-6 torr is pretty normal. Using a nitrogen back fill and a LN trap on the DP might help speed things up.
Good Luck, Bill
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Title-Subject: [Microscopy] [Filtered] SEM: Minimum Vacuum for Tungsten Filament
Question: We are currently experiencing vacuum issues which are causing tungsten oxide growth and premature failure of the filament while operating our JEOL 5900. One of the JEOL technicians has suggested that 5x10-4 torr is acceptable, but I find this hard to believe. The issue we are dealing with is extraordinarily slow pumping times -- 1 hour to obtain 3x10-5 torr and a max vacuum of 5x10-6 torr after 3-4 hours. We are a high volume lab, and so I wish to know what the minimum vacuum I need without seriously diminishing filament life, in order to maximize the workload.
Sincerely,
Cavin T. F. Mooers EM Facility Manager Vitreous State Laboratory The Catholic University of America Hannan Hall ñ Rm 433 Washington, D.C. 20064 (202) 319-6237 (Office) (202) 319-5346 (Lab) (202) 319-4469 (Fax)
No, I had not considered Ir. My basic problem is that I am getting more specimens that have Pt, W, Ta, Si, O, N, Mo, and C (not necessarily all of these elements at the same time!). A typical problem is a Au/Pd coated specimen that has large or small amounts of Pt. Sputter coating with Pt would not be smart, so I use Au/Pd. However, the Pt, when trace or thin, is right next to Au Z-wise. And my 50-70A Au/Pd coating shows up about the height in EDS as does the Pt.
The EDAX Genesis "knows" that it is there via the HPD sanity check. I was thinking of using Os to get further Z distance from the Pt and also from W. The pile up convolution is at the M alpha series for Pt and Au. This can be overcome by increasing KV to 20KV rather than the 10-15KV I usually use. But the specimens sometimes don't like the higher KV and volumetric interaction also can be an issue. So I figured that if I could keep in the M alpha region and put more Z distance between elements, that would be a good move.
I looked at the Emitech you suggested. It is a monster unit! All I need is a small desktop unit for pin stubs. That drew me to the Os unit. But, are you indicating that Os coated specimens will oxidize similar to Cr coating? Not good. perhaps there is no one good answer.
gary g.
At 12:31 PM 1/19/2004, you wrote: } Gary: have you considered Iridium coating? Ir is a noble metal so no } oxidation } will happen. It's also less worrisome than Os tetroxide if you're not used to } using it. One of my labs uses it and has found no noticeable artifacts at } 500KX } SEM. They use Ir foil in an ion-beam sputtering unit and also have an Ir } target in } an Emitech sputter coater (http://www.empdirect.com/k675.html). I think } Ted Pella } also has an Ir option. } } Gary Gaugler wrote: } } } -- } } } } Hello all: } } } } I am considering trying to budget for an } } Osmium-based specimen coater to augment (not replace) } } my Denton Desk II Au/Pd and Pt coater. There } } are some specimens that are going to be taken } } at high mag (250KX-450KX) and should not show } } coating artifacts (major or minor). The Os } } coater looks like a good option. Cr is out } } due to short life span based on rapid oxidation. } } Is the same true for Os? } } } } I would appreciate feedback from Os coater users } } and suppliers. Off-list of course. } } } } tnx, } } gary g. } } -- } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Becky Holdford (r-holdford-at-ti.com) } 972-995-2360 } 972-648-8743 (pager) } SC Packaging FA Development } Texas Instruments, Inc. } Dallas, TX } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 18:30:38 2004
Hi, I would like to stain Biofilm polysaccharide on human specimens with Ruthenium red. How should I get my specimens ready for SEM? dehydration process before or after the stain? 0.05% Ruthenium red in gluteraldehyde or Formalin? what should I expect to look for under SEM? any interferences or false positives with human substances? Appreciate any input. Thanks, Jose
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 19 19:55:52 2004
Formatex, a Spanish Research Center, in association with Kluwer Academic Publishers, is now preparing the Second number of the Formatex Microscopy book series, with the preliminary title of "Current Issues in Multidisciplinary Microscopy Research and Education". You can see the contents of the first number published in 2003 in http://www.formatex.org/micro2002/callforpaper.htm
Website of the 2004 edition is http://www.formatex.org/micro2003/callforpaper2.htm
This second number of the series is committed to giving an overview of the state of the art as well as upcoming trends, and to promoting discussion about scientific, technological and educational aspects of Microscopy in both the biological/biomedical and physical/chemical sciences. Although all types of papers are a priori accepted (research articles, reviews, case studies, etc.), priority will be given to those which clearly emphasize the scientific/technological/pedagogical results, as well as those making comparative discussions of two or more microscopy techniques or showing the complementarity of microscopy techniques with other techniques. For this second number, "educationally-oriented" and mini-review papers are specially welcome, although also "regular" research papers are accepted.
If you are interested in participating in this edition submitting a (technical, scientific, educational, introductory...) chapter related to microscopy, please see the website for details.
As you may see from the Call for Papers' website, the deadline for chapter submission is MARCH 15TH. Early submission of a short abstract of your chapter proposal is appreciated in order to allow potencial authors to know what other authors will write about for the book and avoid contents duplications.
We hope that you find this new approach to microscopy issues interesting and we hope to hear from you/your team for this and/or future editions.
If you any enquiry or suggestion about this volume, please contact us.
Best wishes from Spain.
José Antonio Mesa Gonzalez Editorial Assistant Formatex Research Center C / Encarnacion, 3 1ºE 06001 Badajoz SPAIN Phone/Fax: +34 924258615 Email: micro-at-formatex.org
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 20 02:10:27 2004
Many month ago I have asked for advise regarding a problem with LaB6 filament. Well, finally we decided to switch for W filament. So, the W filament is working in vacuum proper for LaB6 one (ion pump). Now is almost one year (maybe 10 month) and still is working.
Talking about LaB6 problem, during this time we have been observing behaviour of our microscope and it seems that the problem with LaB6 was caused be emergancy shut down of high voltage which was caused probably by wrong signal from dirty Penning gauge. This shut down caused a deposition of a layer (prabably LaB6) on the outside surface of Whenelt cup and this was our problem.
Best regards,
Witold Zielinski
* Date sent: Mon, 19 Jan 2004 16:58:11 -0500 *From: William J Mushock {wim5-at-lehigh.edu} *Organization: Lehigh University *To: microscopy-at-ns.microscopy.com *Subject: [Microscopy] viaWWW: SEM: Minimum Vacuum for Tungsten Filament
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Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
} I have been given two samples and asked to determine } if they are concrete, sandstone, or some mix. } One sample is light gray, and I suspect may be } mortar rather than concrete. The other sample has a slight } rust/red hue and contains appreciably more calcium. } ...
Natural and sedimentary sandstone is likely to run a gamut in composition of clasts and cement (the glue between the grains). One the other hand, I should think concrete would be predominantly calcium sulphate (Portland Cement), but containing clasts from a natural source.
I not been working with TEM diffraction or image simulations programs for a few years now and am wondering what programs are now available and widely used. I do not know which operating system I will be using for the analysis, so any advice will be appreciated.
Thank you for your help. Amy
Amy R. Ross Los Alamos National Laboratory
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 20 09:44:12 2004
} -----Original Message----- } From: Chris Salter } Sent: Tuesday, January 20, 2004 11:11 AM } To: michael shaffer } } No. The mechanism the setting of hydraulic cements (those based on } Portland cement) have nothing to do with Calicum Sulphate. They work on } the hydration of CaAlSilicates - fine tubules grow out from each cement } grain and mesh with those from surrounding grains to give the initial } set, thereafter various other chemical changes occur strengthen the } bonds. The aluminium is essential for thise mechanism to work. } Plasters work by the mechanism you are thinking of, but } plaster is far } weaker than cement or lime mortarand plaster is water soluble. } } } Thanx for the clarification ... but isn't the primary process for the } } hardening of cement the cystallization of CaSO4, i.e.: } } } } CaSO4 (powder) + water =} CaSO4(H2O)n (??) } } } } cheerios ... shAf :o) } } Avalon Peninsula, Newfoundland } } www.micro-investigations.com } } } } } } } } } } } -- } Chris Salter, } Oxford Materials Characterisation Service, } Oxford University Begbroke Science Park, } Sandy Lane, Yarnton, Oxford, OX5 1PF } Tel 01865 283722, EPMA 283741, Mobile 07776031608 } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 20 10:28:34 2004
We are just in the process of buying a FEG-SEM and have been tempted by the vendors to add a STEM-detector to use for TEM imaging of normal tissue sections up to around 40 - 50 kX. This would mean that we could skip our old (17 years) TEM, which would save us money and trouble. I would be very happy for some input from the list on a few issues around this. Is it a good idea to replace a 'proper' TEM with a STEM-detector? What are the limitations with such a setup? The FEG-SEM will run at max 30kV, which will obviously put a limitation to the resolution, but the images we have seen (of our own samples) do not seem to suffer as much as we had expected. Indeed, one of the companies even did the test images at 10 kV, saying that it was the 'best' setting for that kind of images. How is the imaging actually done?
thanks for any views, Stefan +++++++++++++++++++++++++++++++++++++++++++++++++++++++ Dr Stefan Gunnarsson Evolutionsbiologiskt Centrum Evolutionary Biology Centre Enheten för biologisk strukturanalys Microscopy and Imaging Unit Norbyvägen 18A SE75236 Uppsala, Sweden Tel & Fax: +46 - 18471 2638 +++++++++++++++++++++++++++++++++++++++++++++++++++++++
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 20 11:31:14 2004
I am also very interested in the development of this area and seeking to broaden my knowledge. I believe there are some interesting opportunities for SEM-Based STEM in biological applications. However, the utility is yet to be proven (at least I am not aware of a large number of results). So, I would strongly suggest you obtain results on your samples. I don't believe you can say the 30kV SEM-based STEM will replace a quality cryo TEM. But I believe there are some interesting possibilities that need to be explored.
Currently, biological EM imaging does not present challenges in terms of resolution, but rather in terms of contrast and beam induced damage. A low voltage SEM-based STEM image will provide relatively high contrast due to the low voltage. I am aware of some very promising results on biological materials that even challenge zero loss imaging in terms of contrast. A sufficiently high angle STEM will also eliminate diffaction contrast, which is more ideal for tomographic applications (may not be an issue for your samples).
STEM mode on a SEM-based STEM also provides the best resolution mode on the tool. You should be able to achieve subnanometer resolution. The effect of beam broadening, etc., on your particular sample may degrade that ideal performance specification.
The question remaining (to me) is what exactly can be achieved with a 30kV system and what is the beam damage? I assume you'd have a cryo transfer type system. Again, I believe the best option is to see some results on your samples.
I'd be very intersted in your final assessment. Good luck.
Regards, Ed
----- Original Message ----- } From: "Stefan Gunnarsson" {Stefan.Gunnarsson-at-ebc.uu.se} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Tuesday, January 20, 2004 8:38 AM
Chris-
Based on your chemical data neither sample looks like a sandstone to me. A sandstone typically contains a lot of quartz (SiO2), and also contains other minerals such as feldspar, calcite, pyrite and clay minerals. I would expect a lot more aluminum to combine with the Ca and Si to make feldspar and/or clay minerals. Calcite (CaCO3) is a typical mineral that binds the particles in a sandstone (we geologists call this a "cement") and I do not see any Carbon in either analysis. Can you detect carbon with your instrument or not?
Another method you can use to determine the mineralogy of your samples is XRD (X-ray Diffraction). Yes, this is what I do but you need to use the right tool for the job. It does not matter to me if you use me or another lab. XRD will help you get the answer a lot quicker and with a lot more certainty.
Contact me off list if you have any questions.
Sincerely, James Talbot
K/T GeoServices, Inc. Bulk and Clay Mineralogy by X-ray Diffraction www.ktgeo.com (940) 597-9076
At 10:18 AM 1/19/04 -0500, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Jan 20 12:34:17 2004
You can find distributors for Dow Corning products on their website. http://www.dowcorning.com
Choose youre product and that page will have an option to search for local suppliers. We use Sylgard 184 to make dissecting surfaces for biological samples.
On Thu, 15 Jan 2004 16:27:37 -0600 Philip Oshel {peoshel-at-wisc.edu} wrote: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America
Larry Ackerman Keck Advanced Microscopy Lab Dept. of Biochemistry & Biophysics University of California San Francisco 600-16th Street, Room S101, Box 2140 San Francisco, CA 94158 (for postal mail use 94143)
415-476-4441 FAX 415-514-4142
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 20 16:18:39 2004
I agree with James Talbot's opinion that X-ray diffraction would probably give a definitive answer quite quickly. I think you might be able to get a pretty good idea by taking spot analyses or an x-ray map of your two samples. (It is what I do - on concrete.) Concrete or mortar is such a heterogeneous mess that I would not get much out of an overall analysis. However, if I can probe the cement paste and find Ca and Si and O, I can suppose it to be Portland cement. There might also be unreacted cement grains. You should also find substantial amounts of fine aggregate (i.e., sand) used to make the mortar. Those could be of most any composition, and often are. I see calcite, quartz, feldspars and more. Because cement and concrete are such hodge-podge mixtures, I often choose to do an x-ray map rather than to probe every point in the image by hand.
But back to x-ray diffraction, even if you can get chemistry from the SEM, you may still want to use XRD for a more definitive answer about the phases present. I use an SEM to solve a lot of problems, but it has its limitations. There is no substitute for diffraction or thermal analysis or FTIR if the samples require it.
Warren
At 09:18 AM 1/19/2004, you wrote:
} You raise questions on some of the perplexing results from this job. While } I am well aware of the heterogeneous nature of the samples, a summary of } the "bulk" analysis results yielded (approx.): } Lt. gray sample; O 38%, Si 40%, Ca 20%, with Mg, Al, and S also present. } Rust/Red sample; O 48%, Si 4%, Ca 47%, with Mg, Al, S, and Fe also present. } } A small piece from each sample was sanded through 320 grit paper to get a } fresh, flat surface for the EDS analyses. } } Now, neither sample is known to be concrete nor sandstone. Neither sample } contains any aggregate material. The discrete particles present in the } light gray sample are more populous and twice the size of the particles in } the red material. There is no color banding in either sample, which could } be indicative of the stratified layers in a sandstone. } } I may be making this harder than it has to be, but I am suspecting that } neither sample is sandstone.
------------------------------------------- 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
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"Now, neither sample is known to be concrete nor sandstone. ..." Well that doesn't narrow it down any. According to my limited resource; cement would have } 3x more Ca than Si; and in sandstone the ratio of Si to Ca is expected to be much greater than double (what you have). I would guess that neither sample is sandstone.
Perhaps a geologist could guide you further. ED
holpc-at-firstenergycorp.com 01/19/04 10:18 AM
To ekomarnicki-at-MacDermid.com cc Microscopy-at-msa.microscopy.com Subject Re: Re: SEM/LM of concrete vs. sandstone
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You raise questions on some of the perplexing results from this job. While I am well aware of the heterogeneous nature of the samples, a summary of the "bulk" analysis results yielded (approx.): Lt. gray sample; O 38%, Si 40%, Ca 20%, with Mg, Al, and S also present. Rust/Red sample; O 48%, Si 4%, Ca 47%, with Mg, Al, S, and Fe also present.
A small piece from each sample was sanded through 320 grit paper to get a fresh, flat surface for the EDS analyses.
Now, neither sample is known to be concrete nor sandstone. Neither sample contains any aggregate material. The discrete particles present in the light gray sample are more populous and twice the size of the particles in the red material. There is no color banding in either sample, which could be indicative of the stratified layers in a sandstone.
I may be making this harder than it has to be, but I am suspecting that neither sample is sandstone.
ekomarnicki-at-MacDe rmid.com To: holpc-at-firstenergycorp.com cc: Microscopy-at-msa.microscopy.com 01/19/2004 09:13 Subject: [Microscopy] Re: SEM/LM of concrete vs. AM sandstone
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Chris, I'm not a geologist or a cement expert, but concrete and mortar have cement as part of their make-up (ingredient). So you probably should start out comparing sandstone and cement. Commonly, concrete is cement with crushed stones (aggregates) in it. You don't mention having EDS at your disposal. Having EDS spectra obtained on your sample, I would expect higher Si concentration in the sandstone. and higher Ca conc. in the cement. Drop/spots tests may not reveal conc. differences. You may also want to run standards (i. e. Portland Cement, etc.) with your samples for positive confimation. Or even call a cement company and find out how they analyze their products. Hope this helps.
The Materials Handbook, 13th ed. has some basic (starting) info on all three materials. ED
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One and all,
I have been given two samples and asked to determine if they are concrete, sandstone, or some mix. One sample is light gray, and I suspect may be mortar rather than concrete. The other sample has a slight rust/red hue and contains appreciably more calcium. Both samples contain silica, and both test positive for calcium carbonate by the acid drop test, with the light gray sample showing a yellow residue from this test.
Hours of reading test procedures for Portland cement and concrete, coupled with internet searches have left me wondering if I can draw a conclusion based on a few grams of sample. I would appreciate any suggestions about what to read or how to make this kind of determination.
Thanks,
Chris Holp FirstEnergy Corp. Beta Labs HolpC-at-firstenergycorp.com
----------------------------------------- The information contained in this message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately, and delete the original message.
----------------------------------------- The information contained in this message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately, and delete the original message.
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 20 19:19:47 2004
Title-Subject: [Microscopy] [Filtered] Applications for a wide field of view confocal microscope
Question: Hello All,
We are assessing applications for a new type of confocal microscope. The microscope is like a confocal scanning laser microscope, but with ten times the field of view (at the same resolution).
The instrument can be used to image large samples, with a field of view of 1cm at submicron resolution. The instrument can be used in room light and can be highly automated. One of the applications for this microscope is to image tissue samples. In this application, some advantages include:
Images entire biopsy specimens in a single scan (no tiling) Fast scanning ñ scans 10mmX10mm -at- 2 micron in 2 minutes Simultaneous acquisition of 2-fluorophores Scan area is 70mm x 22mm
We are compiling a list of potential applications for this technology (in any sector). Can you think of any specific applications for this technology where the large field of view and submicron resolution would be of particular use?
Thank you very much for your time and help, Best wishes
Hitesh Jain, B.Sc., M.F.C Senior Research Analyst VISTA - The Canadian Centre for Science and Technology Solutions Brock University
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You raise questions on some of the perplexing results from this job. While I am well aware of the heterogeneous nature of the samples, a summary of the "bulk" analysis results yielded (approx.): Lt. gray sample; O 38%, Si 40%, Ca 20%, with Mg, Al, and S also present. Rust/Red sample; O 48%, Si 4%, Ca 47%, with Mg, Al, S, and Fe also present.
A small piece from each sample was sanded through 320 grit paper to get a fresh, flat surface for the EDS analyses.
Now, neither sample is known to be concrete nor sandstone. Neither sample contains any aggregate material. The discrete particles present in the light gray sample are more populous and twice the size of the particles in the red material. There is no color banding in either sample, which could be indicative of the stratified layers in a sandstone.
I may be making this harder than it has to be, but I am suspecting that neither sample is sandstone.
ekomarnicki-at-MacDe rmid.com To: holpc-at-firstenergycorp.com cc: Microscopy-at-msa.microscopy.com 01/19/2004 09:13 Subject: [Microscopy] Re: SEM/LM of concrete vs. AM sandstone
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Chris, I'm not a geologist or a cement expert, but concrete and mortar have cement as part of their make-up (ingredient). So you probably should start out comparing sandstone and cement. Commonly, concrete is cement with crushed stones (aggregates) in it. You don't mention having EDS at your disposal. Having EDS spectra obtained on your sample, I would expect higher Si concentration in the sandstone. and higher Ca conc. in the cement. Drop/spots tests may not reveal conc. differences. You may also want to run standards (i. e. Portland Cement, etc.) with your samples for positive confimation. Or even call a cement company and find out how they analyze their products. Hope this helps.
The Materials Handbook, 13th ed. has some basic (starting) info on all three materials. ED
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One and all,
I have been given two samples and asked to determine if they are concrete, sandstone, or some mix. One sample is light gray, and I suspect may be mortar rather than concrete. The other sample has a slight rust/red hue and contains appreciably more calcium. Both samples contain silica, and both test positive for calcium carbonate by the acid drop test, with the light gray sample showing a yellow residue from this test.
Hours of reading test procedures for Portland cement and concrete, coupled with internet searches have left me wondering if I can draw a conclusion based on a few grams of sample. I would appreciate any suggestions about what to read or how to make this kind of determination.
Thanks,
Chris Holp FirstEnergy Corp. Beta Labs HolpC-at-firstenergycorp.com
----------------------------------------- The information contained in this message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately, and delete the original message.
----------------------------------------- The information contained in this message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately, and delete the original message.
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 20 21:17:50 2004
I am intrigued by the similarity between your problem and a similar problem confronting the Mars Rover scientists. They are hoping that the landing site is a lake bed and are looking for textures indicative of a sedimentary rock type, such as sandstone. All they have is an optical microscope, an alpha-particle x-ray spectrometer and an extraterrestrial grinder to make a flat surface similar to your tools but significantly more expensive.
Sandstone is defined by texture: grain size, shape and sorting, not chemistry. Probably the last instrument a sedimentary petrologist would use is EDS to define the rock type. Look for rounded grains of quartz in a size range from 0.06 mm to 2.0 mm comprising most of the sample. Larger than that is gravel, smaller is mud.
If you can't see the grains in a hand lens or binocular microscope you may have a sediment with smaller grain size such as a siltstone or mudstone or perhaps cement but not sandstone.
Your best bet is examining the texture in a petrographic thin section - are these still made or am I too old.
Dr. Gordon Nord Senior Scientist Center for Applied Studies of the Environment The Graduate School and University Center City University of New York 365 Fifth Avenue NY NY 10016-4309
On Monday, January 19, 2004, at 10:18 AM, holpc-at-firstenergycorp.com 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 } ----------------------------------------------------------------------- } -------- } } } You raise questions on some of the perplexing results from this job. } While } I am well aware of the heterogeneous nature of the samples, a summary } of } the "bulk" analysis results yielded (approx.): } Lt. gray sample; O 38%, Si 40%, Ca 20%, with Mg, Al, and S also } present. } Rust/Red sample; O 48%, Si 4%, Ca 47%, with Mg, Al, S, and Fe also } present. } } A small piece from each sample was sanded through 320 grit paper to } get a } fresh, flat surface for the EDS analyses. } } Now, neither sample is known to be concrete nor sandstone. Neither } sample } contains any aggregate material. The discrete particles present in the } light gray sample are more populous and twice the size of the } particles in } the red material. There is no color banding in either sample, which } could } be indicative of the stratified layers in a sandstone. } } I may be making this harder than it has to be, but I am suspecting that } neither sample is sandstone. } } } } } } ekomarnicki-at-MacDe } rmid.com To: } holpc-at-firstenergycorp.com } cc: } Microscopy-at-msa.microscopy.com } 01/19/2004 09:13 Subject: [Microscopy] } Re: SEM/LM of concrete vs. } AM sandstone } } } } } } } } } ----------------------------------------------------------------------- } ------- } } 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 } ----------------------------------------------------------------------- } -------- } } } Chris, I'm not a geologist or a cement expert, but concrete and mortar } have cement as part of their make-up (ingredient). So you probably } should } start out comparing sandstone and cement. Commonly, concrete is cement } with crushed stones (aggregates) in it. You don't mention having } EDS at } your disposal. Having EDS spectra obtained on your sample, I would } expect } higher Si concentration in the sandstone. and higher Ca conc. in the } cement. Drop/spots tests may not reveal conc. differences. You may } also } want to run standards (i. e. Portland Cement, etc.) with your samples } for } positive confimation. Or even call a cement company and find out how } they } analyze their products. Hope this helps. } } The Materials Handbook, 13th ed. has some basic (starting) info on all } three materials. ED } } Ed Komarnicki } Sr. Analytical Chemist } MacDermid Inc. } } } } } } holpc-at-firstenergycorp.com } 01/19/04 08:09 AM } } To } Microscopy-at-msa.microscopy.com } cc } } Subject } SEM/LM of concrete vs. sandstone } } } } } } } } } ----------------------------------------------------------------------- } ------- } } 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 } ----------------------------------------------------------------------- } -------- } } } One and all, } } I have been given two samples and asked to determine if they are } concrete, } sandstone, or some mix. One sample is light gray, and I suspect may be } mortar rather than concrete. The other sample has a slight rust/red hue } and } contains appreciably more calcium. Both samples contain silica, and } both } test positive for calcium carbonate by the acid drop test, with the } light } gray sample showing a yellow residue from this test. } } Hours of reading test procedures for Portland cement and concrete, } coupled } with internet searches have left me wondering if I can draw a } conclusion } based on a few grams of sample. I would appreciate any suggestions } about } what to read or how to make this kind of determination. } } Thanks, } } Chris Holp } FirstEnergy Corp. } Beta Labs } HolpC-at-firstenergycorp.com } } } } ----------------------------------------- } The information contained in this message is intended only for the } personal and confidential use of the recipient(s) named above. If the } reader of this message is not the intended recipient or an agent } responsible for delivering it to the intended recipient, you are hereby } notified that you have received this document in error and that any } review, dissemination, distribution, or copying of this message is } strictly prohibited. If you have received this communication in error, } please notify us immediately, and delete the original message. } } } } } } } } } } } ----------------------------------------- } The information contained in this message is intended only for the } personal and confidential use of the recipient(s) named above. If the } reader of this message is not the intended recipient or an agent } responsible for delivering it to the intended recipient, you are } hereby notified that you have received this document in error and that } any review, dissemination, distribution, or copying of this message is } strictly prohibited. If you have received this communication in error, } please notify us immediately, and delete the original message. } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 04:53:33 2004
I think you would really need to compare pictures from the same or similar samples. Ideally you would also need to see the process of producing images on the STEM because I suspect time and convenience for routine specimens would also be important.
I use a fairly old (} 10 years) Hitachi H7000 TEM with STEM and SEM facilities and it always takes longer to achieve a good STEM image than a TEM image. The STEM is useful for thick, low contrast specimens or x-ray analysis but for routine specimens the TEM will always be quicker and more convenient. Other issues may include whether it will take longer to train other users or more supervision would be needed with the STEM than with a TEM.
It may well be that modern SEM/STEM combinations are much improved (certainly resolution for a FEG should be much better) but it's worth considering how practical the STEM will be for routine TEM specimens. This will be greatly affected by the number and type of routine TEM specimens you handle.
I hope this helps.
Malcolm
Malcolm Haswell e.m. unit Chemispec School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: principe {eprincipe01-at-hotmail.com}
INVITATION
(1st Announcement)
College of Medicine & Health Sciences
Pathology Department
Electron Microscopy Unit
In collaboration with
Center for Community Service & Continuing Education
Invites you to attend
“OMAN FIRST ELECTRON MICROSCOPY WORKSHOP”
March 6 – 9, 2004
Venue: Lecture Theater 1, Sultan Qaboos University
Opening Ceremony: Saturday March 6 2004 at 9:00 a.m.
Tentative Program Schedule:
Day (1) Saturday, 6 March 2004
8:00-9:00 Registration, Coffee
9:00-9:45 Opening Ceremony
9:45-10:00 Break
10:00-11:00 Lecture
11:00-12:00 Lecture
12:00-12:30 Tour around EM Lab
12:30-14:00 Lunch
13:00-17:30 Sample preparation for TEM
Day (2) Sunday, 7 March 2004
8:30-9:30 Lecture
9:30-10:30 Lecture
10:30-11:00 Break / Exhibition
11:00-12:00 Lecture
12:00-13:00 Lunch
13:00-17:00 Sample preparation for SEM
19:00- 21:00 Workshop Dinner
Day (3) Monday, 8 March 2004
8:30-9:30 Lecture
9:30-10:30 Lecture
10:30-11:00 Break / Exhibition
11:00-12:00 Lecture
12:00-13:00 Lunch
13:00-17:00 Operation & Maintenance of Electron Microscopes
Day (4) Tuesday, 9 March 2004
8:30-9:30 Lecture
9:30-10:30 Lecture
10:30-11:00 Break / Exhibition
11:00-14:00 Operation & Maintenance of Electron Microscopes
14:00-15:00 Lunch
17:00-21:00 Tour - Muscat area
For registration and full details about the workshop please visit our web site address:
http://www.squ.edu.om/med/NEWS/med/index.htm
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 08:03:34 2004
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Dear Microscopists
EDTA decalcification can be carried out on well-fixed specimens, including corals, in a laboratory (tempreature-controlled) microwave in a fraction of the time needed on the bench at room temperature. Stirring during the process is important to keep fresh decalcifying agent at the specimen at all times.
best regards, Steven Slap
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 08:03:22 2004
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Dear Microscopists
EDTA decalcification can be carried out on well-fixed specimens, including corals, in a laboratory (tempreature-controlled) microwave in a fraction of the time needed on the bench at room temperature. Stirring during the process is important to keep fresh decalcifying agent at the specimen at all times.
best regards, Steven Slap
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 08:39:48 2004
The simplest technique is to have petrographic thin sections made (there are several labs that specialize in this service at very reasonable rates and turn-around times) and to use polarized light microscopy (PLM). There are several texts available for both sedimentary petrography (107 listed at Amazon) or cement petrography (101 listed at Amazon). My background is in geology, but I haven't looked at any rocks for a couple of years now, and I have never compared a sandstone to a cement. However, if a client brought a similar problem to my facility, PLM is the technique I would use.
Not to discredit any of the previous techniques (XRD, SEM/EDS, etc.) but PLM is the best way to go with this problem (in my opinion). Especially if you aren't sure if you are looking at sandstone or cement or a third type of rock/cement-like material, a few minutes with a PLM, and a couple of good references one can provide a definite answer.
Bryan Bandli
----- Original Message ----- } From: "Warren E Straszheim" {wesaia-at-iastate.edu} To: {Microscopy-at-msa.microscopy.com} Sent: Tuesday, January 20, 2004 5:26 PM Subject: [Microscopy] Re: SEM/LM of concrete vs. sandstone
} } } -------------------------------------------------------------------------- ---- } 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 agree with James Talbot's opinion that X-ray diffraction would probably } give a definitive answer quite quickly. I think you might be able to get a } pretty good idea by taking spot analyses or an x-ray map of your two } samples. (It is what I do - on concrete.) Concrete or mortar is such a } heterogeneous mess that I would not get much out of an overall analysis. } However, if I can probe the cement paste and find Ca and Si and O, I can } suppose it to be Portland cement. There might also be unreacted cement } grains. You should also find substantial amounts of fine aggregate (i.e., } sand) used to make the mortar. Those could be of most any composition, and } often are. I see calcite, quartz, feldspars and more. Because cement and } concrete are such hodge-podge mixtures, I often choose to do an x-ray map } rather than to probe every point in the image by hand. } } But back to x-ray diffraction, even if you can get chemistry from the SEM, } you may still want to use XRD for a more definitive answer about the phases } present. I use an SEM to solve a lot of problems, but it has its } limitations. There is no substitute for diffraction or thermal analysis or } FTIR if the samples require it. } } Warren } } At 09:18 AM 1/19/2004, you wrote: } } } You raise questions on some of the perplexing results from this job. While } } I am well aware of the heterogeneous nature of the samples, a summary of } } the "bulk" analysis results yielded (approx.): } } Lt. gray sample; O 38%, Si 40%, Ca 20%, with Mg, Al, and S also present. } } Rust/Red sample; O 48%, Si 4%, Ca 47%, with Mg, Al, S, and Fe also present. } } } } A small piece from each sample was sanded through 320 grit paper to get a } } fresh, flat surface for the EDS analyses. } } } } Now, neither sample is known to be concrete nor sandstone. Neither sample } } contains any aggregate material. The discrete particles present in the } } light gray sample are more populous and twice the size of the particles in } } the red material. There is no color banding in either sample, which could } } be indicative of the stratified layers in a sandstone. } } } } I may be making this harder than it has to be, but I am suspecting that } } neither sample is sandstone. } } ------------------------------------------- } 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 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 09:31:46 2004
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Chris, I'm not a geologist or a cement expert, but concrete and mortar have cement as part of their make-up (ingredient). So you probably should start out comparing sandstone and cement. Commonly, concrete is cement with crushed stones (aggregates) in it. You don't mention having EDS at your disposal. Having EDS spectra obtained on your sample, I would expect higher Si concentration in the sandstone. and higher Ca conc. in the cement. Drop/spots tests may not reveal conc. differences. You may also want to run standards (i. e. Portland Cement, etc.) with your samples for positive confimation. Or even call a cement company and find out how they analyze their products. Hope this helps.
The Materials Handbook, 13th ed. has some basic (starting) info on all three materials. ED
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
holpc-at-firstenergycorp.com 01/19/04 08:09 AM
To Microscopy-at-msa.microscopy.com cc
Subject SEM/LM of concrete vs. sandstone
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One and all,
I have been given two samples and asked to determine if they are concrete, sandstone, or some mix. One sample is light gray, and I suspect may be mortar rather than concrete. The other sample has a slight rust/red hue and contains appreciably more calcium. Both samples contain silica, and both test positive for calcium carbonate by the acid drop test, with the light gray sample showing a yellow residue from this test.
Hours of reading test procedures for Portland cement and concrete, coupled with internet searches have left me wondering if I can draw a conclusion based on a few grams of sample. I would appreciate any suggestions about what to read or how to make this kind of determination.
Thanks,
Chris Holp FirstEnergy Corp. Beta Labs HolpC-at-firstenergycorp.com
----------------------------------------- The information contained in this message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately, and delete the original message.
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 13:23:19 2004
Hi to all: Attached to our HItachi H600 TEM, we have a Kevex Delta EDX system that has gone out. Specifically, smoke was seen coming out of the monitor shortly before it was last shut off with no image. The system was working fine until this event. We got a replacement monitor that has 5 inputs (RGBHV), but we still get no image. Is there a special type of monitor needed for these old systems, or should we be looking deeper into the system for other problems? The old monitor was an Electrohome ECM 1311U, Model 38-D051MA-UU. Off-list replies OK. thx Mark Floyd Forensic Analytical Hayward, CA msf-at-forensica.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 13:45:55 2004
Does anyone happen to have the error codes for the Agfa scanners? We are having problems in scanning negatives, although the scans of reflective materials are working fine.
We get back an error code saying the "scanner has a problem" referencing code : F0000400:5
The Duoscan HiD model is hooked up to a Mac, and was working faithfully for years. Reloading the software from the original CD has helped us to get the error code rather than a freeze of the machine, but has not solved the problem.
The Agfa machines are now out of production. I looked online and found part of the manual available, but it doesn't include the pages detailing troubleshooting. Thanks for any help in advance.
DHH -- 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 718 430-8821
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 14:21:51 2004
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You raise questions on some of the perplexing results from this job. While I am well aware of the heterogeneous nature of the samples, a summary of the "bulk" analysis results yielded (approx.): Lt. gray sample; O 38%, Si 40%, Ca 20%, with Mg, Al, and S also present. Rust/Red sample; O 48%, Si 4%, Ca 47%, with Mg, Al, S, and Fe also present.
A small piece from each sample was sanded through 320 grit paper to get a fresh, flat surface for the EDS analyses.
Now, neither sample is known to be concrete nor sandstone. Neither sample contains any aggregate material. The discrete particles present in the light gray sample are more populous and twice the size of the particles in the red material. There is no color banding in either sample, which could be indicative of the stratified layers in a sandstone.
I may be making this harder than it has to be, but I am suspecting that neither sample is sandstone.
ekomarnicki-at-MacDe rmid.com To: holpc-at-firstenergycorp.com cc: Microscopy-at-msa.microscopy.com 01/19/2004 09:13 Subject: [Microscopy] Re: SEM/LM of concrete vs. AM sandstone
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Chris, I'm not a geologist or a cement expert, but concrete and mortar have cement as part of their make-up (ingredient). So you probably should start out comparing sandstone and cement. Commonly, concrete is cement with crushed stones (aggregates) in it. You don't mention having EDS at your disposal. Having EDS spectra obtained on your sample, I would expect higher Si concentration in the sandstone. and higher Ca conc. in the cement. Drop/spots tests may not reveal conc. differences. You may also want to run standards (i. e. Portland Cement, etc.) with your samples for positive confimation. Or even call a cement company and find out how they analyze their products. Hope this helps.
The Materials Handbook, 13th ed. has some basic (starting) info on all three materials. ED
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One and all,
I have been given two samples and asked to determine if they are concrete, sandstone, or some mix. One sample is light gray, and I suspect may be mortar rather than concrete. The other sample has a slight rust/red hue and contains appreciably more calcium. Both samples contain silica, and both test positive for calcium carbonate by the acid drop test, with the light gray sample showing a yellow residue from this test.
Hours of reading test procedures for Portland cement and concrete, coupled with internet searches have left me wondering if I can draw a conclusion based on a few grams of sample. I would appreciate any suggestions about what to read or how to make this kind of determination.
Thanks,
Chris Holp FirstEnergy Corp. Beta Labs HolpC-at-firstenergycorp.com
----------------------------------------- The information contained in this message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately, and delete the original message.
----------------------------------------- The information contained in this message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately, and delete the original message.
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 14:32:48 2004
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Ed and Chris,
The difference between sandstones and concretes is usually fairly obvious in a polished thin section viewed under plain and crossed polars. The Atlas of sedimentary rocks under the microscope (Adams, MacKenzie and Guilford, 1984, Longmans) gives examples of the commonest types of sedimentary rock. A sandstone is composed of two components the clasts (sand/rock) and the cement (sorry that is the term the geologists use). Either of these components may contain calcium bearing minerals, so the simple presence of a signficiant calcium peak in an EDX spectra may not necessarily prove that the material is a concrete. However, the cement of concrete will not normally show visible crystallinity in the optical microscope whereas many sandstones will. It also allows you to identify the clasts and their size so distinguish between mortar and concrete (if samples is large enough to include representative clasts).
ekomarnicki-at-MacDermid.com 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 } ------------------------------------------------------------------------------- } } Chris, I'm not a geologist or a cement expert, but concrete and mortar } have cement as part of their make-up (ingredient). So you probably should } start out comparing sandstone and cement. Commonly, concrete is cement } with crushed stones (aggregates) in it. You don't mention having EDS at } your disposal. Having EDS spectra obtained on your sample, I would expect } higher Si concentration in the sandstone. and higher Ca conc. in the } cement. Drop/spots tests may not reveal conc. differences. You may also } want to run standards (i. e. Portland Cement, etc.) with your samples for } positive confimation. Or even call a cement company and find out how they } analyze their products. Hope this helps. } } The Materials Handbook, 13th ed. has some basic (starting) info on all } three materials. ED } } Ed Komarnicki } Sr. Analytical Chemist } MacDermid Inc. } } } } } } holpc-at-firstenergycorp.com } 01/19/04 08:09 AM } } To } Microscopy-at-msa.microscopy.com } cc } } Subject } SEM/LM of concrete vs. sandstone } } } } } } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------- } } One and all, } } I have been given two samples and asked to determine if they are } concrete, } sandstone, or some mix. One sample is light gray, and I suspect may be } mortar rather than concrete. The other sample has a slight rust/red hue } and } contains appreciably more calcium. Both samples contain silica, and both } test positive for calcium carbonate by the acid drop test, with the light } gray sample showing a yellow residue from this test. } } Hours of reading test procedures for Portland cement and concrete, coupled } with internet searches have left me wondering if I can draw a conclusion } based on a few grams of sample. I would appreciate any suggestions about } what to read or how to make this kind of determination. } } Thanks, } } Chris Holp } FirstEnergy Corp. } Beta Labs } HolpC-at-firstenergycorp.com } } } } ----------------------------------------- } The information contained in this message is intended only for the } personal and confidential use of the recipient(s) named above. If the } reader of this message is not the intended recipient or an agent } responsible for delivering it to the intended recipient, you are hereby } notified that you have received this document in error and that any } review, dissemination, distribution, or copying of this message is } strictly prohibited. If you have received this communication in error, } please notify us immediately, and delete the original message. } } } } } }
-- Chris Salter, Oxford Materials Characterisation Service, Oxford University Begbroke Science Park, Sandy Lane, Yarnton, Oxford, OX5 1PF Tel 01865 283722, EPMA 283741, Mobile 07776031608
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 15:11:25 2004
A quick question, does anyone known the thermal conductivity/properties of carbon adhesive tabs used to mount SEM samples? The various vendors talk about electrical conductivity but not thermal.
Thanks Scott -- ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 15:14:28 2004
We are studying cell migration, as a result, I am interested in opinions/suggestions on various motion tracking software solutions for the analysis of cell movement. In my limited experience, I found that following fluorescently labelled cells is easier than brighfield cells - simpler to find fiducial points - so any suggestions for both types of studies would be highly appreciated.
Thank you Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 7Queen 30 Bond St. Toronto, ON M5B 1W8 Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 21 15:14:27 2004
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Ed and Chris,
The difference between sandstones and concretes is usually fairly obvious in a polished thin section viewed under plain and crossed polars. The Atlas of sedimentary rocks under the microscope (Adams, MacKenzie and Guilford, 1984, Longmans) gives examples of the commonest types of sedimentary rock. A sandstone is composed of two components the clasts (sand/rock) and the cement (sorry that is the term the geologists use). Either of these components may contain calcium bearing minerals, so the simple presence of a signficiant calcium peak in an EDX spectra may not necessarily prove that the material is a concrete. However, the cement of concrete will not normally show visible crystallinity in the optical microscope whereas many sandstones will. It also allows you to identify the clasts and their size so distinguish between mortar and concrete (if samples is large enough to include representative clasts).
ekomarnicki-at-MacDermid.com 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 } ------------------------------------------------------------------------------- } } Chris, I'm not a geologist or a cement expert, but concrete and mortar } have cement as part of their make-up (ingredient). So you probably should } start out comparing sandstone and cement. Commonly, concrete is cement } with crushed stones (aggregates) in it. You don't mention having EDS at } your disposal. Having EDS spectra obtained on your sample, I would expect } higher Si concentration in the sandstone. and higher Ca conc. in the } cement. Drop/spots tests may not reveal conc. differences. You may also } want to run standards (i. e. Portland Cement, etc.) with your samples for } positive confimation. Or even call a cement company and find out how they } analyze their products. Hope this helps. } } The Materials Handbook, 13th ed. has some basic (starting) info on all } three materials. ED } } Ed Komarnicki } Sr. Analytical Chemist } MacDermid Inc. } } } } } } holpc-at-firstenergycorp.com } 01/19/04 08:09 AM } } To } Microscopy-at-msa.microscopy.com } cc } } Subject } SEM/LM of concrete vs. sandstone } } } } } } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------- } } One and all, } } I have been given two samples and asked to determine if they are } concrete, } sandstone, or some mix. One sample is light gray, and I suspect may be } mortar rather than concrete. The other sample has a slight rust/red hue } and } contains appreciably more calcium. Both samples contain silica, and both } test positive for calcium carbonate by the acid drop test, with the light } gray sample showing a yellow residue from this test. } } Hours of reading test procedures for Portland cement and concrete, coupled } with internet searches have left me wondering if I can draw a conclusion } based on a few grams of sample. I would appreciate any suggestions about } what to read or how to make this kind of determination. } } Thanks, } } Chris Holp } FirstEnergy Corp. } Beta Labs } HolpC-at-firstenergycorp.com } } } } ----------------------------------------- } The information contained in this message is intended only for the } personal and confidential use of the recipient(s) named above. If the } reader of this message is not the intended recipient or an agent } responsible for delivering it to the intended recipient, you are hereby } notified that you have received this document in error and that any } review, dissemination, distribution, or copying of this message is } strictly prohibited. If you have received this communication in error, } please notify us immediately, and delete the original message. } } } } } }
-- Chris Salter, Oxford Materials Characterisation Service, Oxford University Begbroke Science Park, Sandy Lane, Yarnton, Oxford, OX5 1PF Tel 01865 283722, EPMA 283741, Mobile 07776031608
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 22 00:45:58 2004
Hi all, Thank you to everyone who answered my query about TEM processing of coral. We now have several methods to try, and the samples won't be wasted.
Cheers,
Lyn Waterhouse Adelaide Microscopy University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 or 8303 5855 Fax: (08) 8303 4356 http://www.adelaide.edu.au/microscopy
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 22 08:20:29 2004
I can't find the web site of this society? SMI (Scientific Manufacturing Industries), 1399 Sixty-fourth Street EMERYVILLE, CAL. 94608
perhaps it has moved?
I am surching for this product micro-re/pettor : 1055A type D
Thank you all
Didier Goux -CENTRE DE MICROSCOPIE ELECTRONIQUE Université de CAEN BASSE NORMANDIE- Campus I - Esplanade de la Paix 14032 CAEN cedex FRANCE Tel : (33) 02.31.56.58.13 - Fax : (33) 02.31.56.56.00 } e-mail: didier.goux-at-unicaen.fr visit our Web page : http://www.microscopie.unicaen.fr/microscopie/
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 22 09:29:48 2004
Dear all, I am trying to get a first rough idea of the cost of an above-screen CCD camera for a TEM, suitable for recording Kikuchi patterns, together with other diffraction patterns and images. I am looking at an above-screen solution as my application requires a largish field of view. It would need to fit in the ports available on a new TEM (yet to be ordered) to be installed at the University of Glasgow (where I shall move to shortly).
If someone wishes to quote for a complete system including software for the indexing of Kikuchi patterns, then I would also find that useful.
Also, generally I would be interested in any software for, or recent work on, the automatic indexing of TEM Kikuchi patterns.
I need this information to help in making suitable estimates for the equipment costings in a research proposal. Since the equipment is for the UK, please give all estimates in British Pounds, where possible.
All the best
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 22 12:20:49 2004
We have a rare mouse coming in and don't want to goof on this one so I wanted some advise.
Does anyone have any recommendations for fixation of neonatal (3 day) mouse cardiac muscle for immuno? I had planned to use 0.5% glutaraldehyde + 3% PAF in 0.1M cacodylate buffer pH 7.4 containing 2mM MgCl, 1mMCaCl2, and 0.25% NaCl (final concentrations).
Do you fix neonatal and adult tissue similarly? If not, what modifications do you make for the adult?
Many thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 22 14:51:45 2004
I have a couple of students who need to look at thin films in cross section in the TEM. Is there a classic reference for preparing cross section samples?
Thanks,
Tom
------------------------------------------------------------------------ --------------------------------------------- Thomas M Murray email: murraytm-at-u.washington.edu Electron Microscopy Center manager Phone: (206)543-2836 Materials Science & Engineering Fax: (206)543-3100 Box 352120 302 Roberts Hall Cell: (425)345-0083 University of Washington Home: (425)742-6819 Seattle, WA 98195
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 22 15:54:16 2004
At 01:00 PM 1/22/2004 -0800, you wrote: } I have a couple of students who need to look at thin films in cross } section in the TEM. Is there a classic reference for preparing cross } section samples?
It's probably bad form to reply to a message asking for "classic references" with a citation to one of my own papers, but I did write something recently on preparing thin film cross sections by tripod polishing. The paper is
{http://dx.doi.org/10.1016/S0304-3991(03)00092-5} "Imaging Individual Atoms Inside Crystals with ADF-STEM" P. M. Voyles, J. L. Grazul, and D. A. Muller, Ultramicroscopy 96, 251 (2003),
one the web at {http://dx.doi.org/10.1016/S0304-3991(03)00092-5} . It contains a complete recipe for tripod polishing modified for HRTEM on blanket thin film samples.
For devices or to achieve very large electron transparent but slightly thicker areas, the original tripod polishing technique described in:
S. J. Klepeis, J. P. Benedict and R. M. Anderson, in: J. C. Bravman (Ed), Materials Research Society Vol. 115, Pittsburgh Pa., 1988, p. 179.
J. Benedict, R. Anderson and S. J. Klepeis, in: R. Anderson, B. Tracy and J. Bravman (Ed), Materials Research Society, Vol. 254, Boston, 1992, p. 121
is better.
Best wishes, Paul Voyles
Paul Voyles Assistant Professor Materials Science and Engineering Department University of Wisconsin - Madison 1509 University Ave. Madison, WI 53706-1595 Voice: (608) 265-6740 Fax: (608) 262-8353 voyles-at-engr.wisc.edu www.engr.wisc.edu/mse/faculty/voyles_paul.html
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 22 17:41:09 2004
There are several excellent references for doing TEM cross sections depending on the method you would like to use. I have listed a few of them below.
Dimpling/Ion Milling Bravman and Sinclair, "The Preparation of Cross Section Specimens for Transmission ELectron Microscopy, Journal of Electron Microscopy Technique 1:53-61 (1984)
McCaffrey and Barna, "Preparation of Cross Sectional TEM Samples for Low Angle Ion Milling, Microscpoy Reasearch and Technique 36:362-367 (1997)
MicroCleaving McCaffrey, "TEM Samples of Semiconductors Prepared by a Small-Angle Cleavage Technique", Materials Research Society Symposium Volume 254 pp 109-120
S.D. Walck, H. Colijn, G. Thompson, "Pre-thinning for FIB TEM Specimen Preparation Using the Small Angle Cleavage Technique", Microscopy and Microanalysis, Vol. 7(2001).
Tripod Polishing J. Benedict, R. Anderson, S. Klepeis, M. Chaker, A Procedure for Cross Sectioning Specific Semiconductor Devices for both SEM and TEM Analysis, ed. R. Anderson, Mat. Res. Soc. Proc. vol 199, Pittsburgh, PA USA. Pp. 189, 1990.
We have a very thorough bibliography on our website which you can also take a look at. If you find anything of interest there, let me know and I will send you a copy. We also have numerous applcaition notes that can be downloaded from the site.
There is also an excellent book you may want to look at titled:
"Progress in Transmission Eelctron Microscopy 1 - Concepts and Techniques". The ISBN number is 3-540-67680-5 and it is available from Springer (http://www.springer.de). The book is also available from South Bay Technology, Inc. at a reduced price.
Chapter 10 in the book it titled "Advanced Techniques in TEM Specimen Preparation". The chapter is edited by Shane Roberts who is the Director of our Applications Laboratory and includes sections written by Dr. John McCaffrey, Dr. Lucille Giannuzzi and Dr. Nestor J. Zaluzec.
Also Volumes 254 and 480 Of the Materials Research Society Proceedings would be a great addition to your library.
I would be pleased to provide other references or advice at any time. Please feel free to contact me or visit our website at www.southbaytech.com and navigate to "Applications Support".
Best regards-
David
DISCLAIMER: As one of the world's leading suppliers of sample preparation equipment and supplies for electron microscopy, South Bay Technology, Inc. has a vested interest in promoting all of these techniques as well as their use.
Tom Murray wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I have a couple of students who need to look at thin films in cross } section in the TEM. Is there a classic reference for preparing cross } section samples? } } Thanks, } } Tom } } ------------------------------------------------------------------------ } --------------------------------------------- } Thomas M Murray email: murraytm-at-u.washington.edu } Electron Microscopy Center manager Phone: (206)543-2836 } Materials Science ? Engineering Fax: (206)543-3100 } Box 352120 302 Roberts Hall Cell: (425)345-0083 } University of Washington Home: (425)742-6819 } Seattle, WA 98195
-- 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.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 22 18:25:12 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a couple of students who need to look at thin films in cross section in the TEM. Is there a classic reference for preparing cross section samples?
Thanks,
Tom
------------------------------------------------------------------------ --------------------------------------------- Thomas M Murray email: murraytm-at-u.washington.edu Electron Microscopy Center manager Phone: (206)543-2836 Materials Science & Engineering Fax: (206)543-3100 Box 352120 302 Roberts Hall Cell: (425)345-0083 University of Washington Home: (425)742-6819 Seattle, WA 98195
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 22 19:06:33 2004
Just to make this interesting thread a little more complete, let me add two wildly different techniques that produce remarkably similar TEM specimens (i.e. uniform, controlled thickness):
1. Diamond knife ultramicrotomy - I list some 20 thin film examples in a slide I use to illustrate the versatility of UM, though some are coatings rather than thin films. (How about diamond on cubic born nitride on single crystal silicon!). The main advantage here is that almost every campus will have ultramicrotomes in various life science departments (Medicine, Biology, etc). Learning the technique for 'hard' materials is a challenge, however, so your films better be fairly soft if you wish immediate results from the life science campus practitioners.
2. Focused ion beam (FIB) - though David noted Lucille Gianuzzi's contribution to the textbook he mentioned, there is still a fair bit of unawareness concerning the versatility of FIB for cross-sections. We have a collaboration with a small FIB company up here in the Great White North, and if you visit their website (www.fibics.com), you will see interesting images of cross-sections through Kodak film and a beverage can label, both of which many might have prejudged to likley have unacceptable levels of ion damage. That might still hold true for high resolution TEM, but these SED images suggest that a lot can still be observed in 'soft' films sectioned via FIB. However, the cost of a FIB is non-trivial, and there are still not that many on campuses.
Tom
Dr. Tom Malis Scientist Advisor Natural Resources Canada Govt. of Canada Phone: 613-995-7358 cell: 613-371-4577 FAX: 613-947-6606 malis-at-nrcan.gc.ca
-----Original Message----- } From: David Henriks To: Tom Murray Cc: Microscopy Listerver Sent: 1/22/2004 5:32 AM
Tom:
There are several excellent references for doing TEM cross sections depending on the method you would like to use. I have listed a few of them below.
Dimpling/Ion Milling Bravman and Sinclair, "The Preparation of Cross Section Specimens for Transmission ELectron Microscopy, Journal of Electron Microscopy Technique 1:53-61 (1984)
McCaffrey and Barna, "Preparation of Cross Sectional TEM Samples for Low Angle Ion Milling, Microscpoy Reasearch and Technique 36:362-367 (1997)
MicroCleaving McCaffrey, "TEM Samples of Semiconductors Prepared by a Small-Angle Cleavage Technique", Materials Research Society Symposium Volume 254 pp 109-120
S.D. Walck, H. Colijn, G. Thompson, "Pre-thinning for FIB TEM Specimen Preparation Using the Small Angle Cleavage Technique", Microscopy and Microanalysis, Vol. 7(2001).
Tripod Polishing J. Benedict, R. Anderson, S. Klepeis, M. Chaker, A Procedure for Cross Sectioning Specific Semiconductor Devices for both SEM and TEM Analysis, ed. R. Anderson, Mat. Res. Soc. Proc. vol 199, Pittsburgh, PA USA. Pp. 189, 1990.
We have a very thorough bibliography on our website which you can also take a look at. If you find anything of interest there, let me know and I will send you a copy. We also have numerous applcaition notes that can be downloaded from the site.
There is also an excellent book you may want to look at titled:
"Progress in Transmission Eelctron Microscopy 1 - Concepts and Techniques". The ISBN number is 3-540-67680-5 and it is available from Springer (http://www.springer.de). The book is also available from South Bay Technology, Inc. at a reduced price.
Chapter 10 in the book it titled "Advanced Techniques in TEM Specimen Preparation". The chapter is edited by Shane Roberts who is the Director of our Applications Laboratory and includes sections written by Dr. John McCaffrey, Dr. Lucille Giannuzzi and Dr. Nestor J. Zaluzec.
Also Volumes 254 and 480 Of the Materials Research Society Proceedings would be a great addition to your library.
I would be pleased to provide other references or advice at any time. Please feel free to contact me or visit our website at www.southbaytech.com and navigate to "Applications Support".
Best regards-
David
DISCLAIMER: As one of the world's leading suppliers of sample preparation equipment and supplies for electron microscopy, South Bay Technology, Inc. has a vested interest in promoting all of these techniques as well as their use.
Tom Murray wrote:
} ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } I have a couple of students who need to look at thin films in cross } section in the TEM. Is there a classic reference for preparing cross } section samples? } } Thanks, } } Tom } } ------------------------------------------------------------------------ } --------------------------------------------- } Thomas M Murray email: murraytm-at-u.washington.edu } Electron Microscopy Center manager Phone: (206)543-2836 } Materials Science ? Engineering Fax: (206)543-3100 } Box 352120 302 Roberts Hall Cell: (425)345-0083 } University of Washington Home: (425)742-6819 } Seattle, WA 98195
-- 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.
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 23 03:16:51 2004
----- Forwarded by ahk/RES/RVN/Haldor Topsoe on 23-01-04 10:24 -----
"Anne-Mette Heie Kjær" To: Tom Murray {murraytm-at-u.washington.edu} {ahk-at-topsoe.dk} cc: bcc: 23-01-04 09:23 Subject: [Microscopy] Re: Reference for TEM cross-section Sample Prep(Document link: ahk)
Hi!
If you have to prepare the cross-section "by hand in the oldfashioned way" - without ultramicrotome or FIB, I would strongly advise you to buy a Gatan cross-sectional kit. It saves you a lot of time.
Best regards Anne-Mette Heie Kjaer
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 23 05:45:55 2004
A few openings are still available for the course on Quantitative Image Analysis that will be taught in Orlando, Florida, March 11 - 12 by John Russ. The course emphasizes practical solutions to imaging problems and includes 2 days of intensive training in current techniques, including an evening open laboratory where participants are encouraged to bring and work with their own images. The course is taught using the Fovea Pro software {http://www.reindeergraphics.com/foveapro} , which participants receive as well as a copy of "The Image Processing Handbook" 4th Edition (CRC Press).
The course syllabus and registration information are available at the Reindeer Graphics website. {http://www.reindeergraphics.com/courses} Further information may be obtained from the website or by contacting Reindeer Graphics directly at courses-at-reindeergraphics.com or by telephone at 828.252.7515. The deadline for registration is February 1st, so act now.
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 23 06:51:32 2004
An early paper on automatic indexing of Kikuchi line patterns was
N. C. Krieger Lassen: "Computerized analysis of Kikuchi patterns" In: Proceedings of the 16th Risoe International Symposium on Materials Science, "Microstructural and Crystallographic Aspects of Recrystallization" Edited by N. Hansen et al. Risoe National Laboratory, Roskilde, Denmark 1995 ISBN 87-550-2088-7 ISSN o907-0079
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: 22. januar 2004 16:37 To: Microscopy Listserver
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (petra.rosner-at-ww.uni-erlangen.de) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, January 23, 2004 at 06:31:59 ---------------------------------------------------------------------------
Email: petra.rosner-at-ww.uni-erlangen.de Name: Petra Rosner
Title-Subject: [Microscopy] [Filtered] MListserver: auto-fluorescence of melanin in fetal human eye
Question: Will the melanin contained in the retinal pigment epithelium and choroid of the fetal human eye absorb the light and prevent visualization of fluorescently labeled endothelial cells, etc. in the choriodal vessels of unbleached whole mount choroidal tissue? If it is possible to visualize the choroidal vessels, what would be the most efficient excitation wave lenght to use? And finally, would a conventional inverted fluorescent scope with a UV and a super high pressure mercury lamp power supply be sufficient for this purpose or is a confocal required?
We are surplusing a cabinet full of old RCA EMU TEM manuals and parts (we've announced this before, but it's for real this time), and wanted to check if anybody has any use for them. We have lots a vacuum tubes, both new and used, miscellaneous parts, like lead glass, a film holder, a lot of new and used filaments, and others, as well as a stack of hardbound manuals. The old kind, with decent binding and illustrations and glossy paper.
If this interests anyone, we will send them for shipping costs. We need the cabinet space for our overflowing supply of researchers' grids.
Thanks,
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 23 08:37:41 2004
I have also experienced the constantly changing guidelines for UA disposal brought up by Tom Phillips. We have gone full circle here from "dilute and pour down the drain", to collecting it with other fixatives for pickup, to collecting it separately for pickup and disposal by the radiation people (along with any pipettes, syringes, etc. that have touched it), and back to dilute and pour. In my experience a major rule change happens about every six months. The problem seems to be that the extremely low level of radioactivity apparently doesn't justify the very high cost of the specialized disposal methods used for "hot" wastes (er, used materials---sorry), and there doesn't seem to be any consensus about how dangerous small amounts of the material actually are. The odd thing to me is that I have heard for many years, rightly or wrongly, that UA is only dangerous when allowed to accumulate in larger quantities or when it is in powder form and can be inhaled. It seems strange then that rules for pickup either seem to require its accumulation or, more rarely, the evaporation of the solvent liquid to return UA to powder form and reduce the collection volume (although I haven't seen that particularly approach required any place where I have worked---so far).
My favorite UA tale comes from another lab I worked in, when we were told by the safety people that we could use UA, but we could not pour it down the sink, we were not allowed to accumulate it, and the safety folks would no longer pick it up because they didn't know what to do with it. What did we do? We kept it large bottles out of sight until the inevitable rule change happened.
Maybe the microscopy community could come up with a consensus recommendation on how to handle this problematic substance in an effort to bring some standardization to its handling. The web of federal, state, and local regulations on hazardous waste probably make this difficult, but we need to try to seal up the crack that UA appears to keep falling through.
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 23 09:28:19 2004
My favorite story: Where I was in graduate school was a small research institute, started around 1900. Over the years, a large set of shelves where chemicals were stored went from communal necessity to historical relic as labs maintained their own supplies. By the time I was there, these shelves were fascinating, containing bottles of colored powder with handwritten labels from Germany or murky gray-black solids labeld odd things like 'hairy tin'. One day, someone looking at the more neglected shelves covering last part of the alphabet, way down near the floor, found a bottle of UA. Handwritten label, radiation detectable with a Geiger counter through the bottle (probably not so pure as today's stuff). Ignored, this stuff for years had led an untroubled existence, but discovered--something had to be done. But what? The institute was attached to a University which was right then being hammered by the NRC for alleged abuses of radiation safety, so they wanted to play by the rules. But the trouble was that the rules demanded that all radioactive compounds be inventoried as soon as they arrived on campus, and fines were levied based on the number of days the compound was on campus but off inventory. There were a lot of days between then and say 1907! The solution was simple but not elegant. It involved some shovels, a bag or two of ready mix, and a big hole in the back of the compound. You can imagine the rest.
The other comment is to point out why UA causes such a regulatory problem. Danger from low doses of anything is hard to study because the sample sizes are too huge. So danger from low doses is extrapolated from high doses. This is obviously uncertain and leads to lots of models, and with the uncertainty, a lot of argument. A useful benchmark for UA is the radiation in the rocks all around us. Probably the diluted UA we use to stain grids has on that order of radiation and adding it the drain poses no more risk than posed by our surroundings. Likewise the small amount of heavy metal in the solution can probably be reduced to the usual ambient levels in tap water by modest dilution. But agreeing to this in a regulatory sense means taking a stand on the low dose end and this is difficult. Though probably wise.
My few decays, Tobias } } } Hi all, } } I have also experienced the constantly changing guidelines for UA } disposal brought up by Tom Phillips. We have gone full circle here from } "dilute and pour down the drain", to collecting it with other fixatives } for pickup, to collecting it separately for pickup and disposal by the } radiation people (along with any pipettes, syringes, etc. that have } touched it), and back to dilute and pour. In my experience a major rule } change happens about every six months. The problem seems to be that the } extremely low level of radioactivity apparently doesn't justify the very } high cost of the specialized disposal methods used for "hot" wastes (er, } used materials---sorry), and there doesn't seem to be any consensus } about how dangerous small amounts of the material actually are. The odd } thing to me is that I have heard for many years, rightly or wrongly, } that UA is only dangerous when allowed to accumulate in larger } quantities or when it is in powder form and can be inhaled. It seems } strange then that rules for pickup either seem to require its } accumulation or, more rarely, the evaporation of the solvent liquid to } return UA to powder form and reduce the collection volume (although I } haven't seen that particularly approach required any place where I have } worked---so far). } } My favorite UA tale comes from another lab I worked in, when we were } told by the safety people that we could use UA, but we could not pour it } down the sink, we were not allowed to accumulate it, and the safety } folks would no longer pick it up because they didn't know what to do } with it. What did we do? We kept it large bottles out of sight until } the inevitable rule change happened. } } Maybe the microscopy community could come up with a consensus } recommendation on how to handle this problematic substance in an effort } to bring some standardization to its handling. The web of federal, } state, and local regulations on hazardous waste probably make this } difficult, but we need to try to seal up the crack that UA appears to } keep falling through. } } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Anthony_McCormick-at-brown.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, January 23, 2004 at 09:01:03 ---------------------------------------------------------------------------
Email: Anthony_McCormick-at-brown.edu Name: Anthony McCormick
Question: We have recently had a window failure on a HPGe x-ray detector for the JEOL 2010 TEM. Upon asking the manufacturer of this detector about repairing the window, their reply was "We can only repair the HPGe detector only if it is converted to a SiLi detector...". Are HPGe detectors no longer available? Are there companies that repair/replace windows on x-ray detectors on equipment that they did not manufacture?
Anthony McCormick Electron Microscope Facility 014 Barus & Holley Brown University Providence, RI 02912 (401) 863-3909
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (swwatkins-at-pitt.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, January 23, 2004 at 11:30:40 ---------------------------------------------------------------------------
Email: swwatkins-at-pitt.edu Name: Simon watkins
Organization: University of Pittsburgh
Title-Subject: [Microscopy] [Filtered] QFM 2k04
Question: Folks,
We would like to announce that enrollment for Quantitative Fluorescence Microscopy 2k04 at the Mount Desert Island Marine Laboratory in Maine is now open. The tremendous success of the course over the last several years has encouraged us to run it again next year, and as it is January it is time to start our enrollment. This is an intensive lab/lecture course, focusing on fluorescence microscopy in all its forms, including widefield, Confocal, Multiphoton, Decon, TIRF, FRET etc, though with an emphasis on live cell methods. We also encourage students to provide their own specimens such that the hands on components of the course are as enriching as possible. Enrollment is highly competitive, so if you think that you or your colleagues would benefit from the course please enroll as soon as possible. The url for the course is http://www.cbi.pitt.edu/qfm/index.html
If you have any specific questions about the course feel free to contact me directly I look forward to hearing from interested applicants soon Thanks Simon
By the way; I am sorry if this is a repost, but I still have not seen my original mailing to the group. Nestor, could you let me know if this has been posted so I avoid future duplications.
Dear List, I have lost track of Doug Dorset's contact info, and the MSA directory did not reveal it. Could someone--preferably Doug--please reply off-list with his email address? TIA. 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 Fri Jan 23 13:52:51 2004
Dear Lists, We have been experiencing an unusual problem on the F30H. At intervals of a few weeks the vacuum measured by the IGP at the top of the FEG (IGP3 for those who know the scope) deteriorates and the FEG is shut off by the computer. Examination of a record of the vacuum shows that IGP3 has a vacuum too good to show above baseline for the first few days--the reading is constant at 10^-12 Pa--then the pressure rises into the range of ~10^-7 Pa, often will briefly get slightly better, then will rather quickly get worse and the FEG will shut off. The boards containing the power supply and the electronics for measuring the vacuum have been replaced, but that did not solve the problem. Both the FEG and IGP are pretty simple devices, and it is hard to think of what could be going wrong. A vacuum leak would not show the observed time course of vacuum readings, and the SF6 that surrounds the outside of the FEG apparatus would give different symptoms. Has anyone seen anything similar? TIA for any help. 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 Fri Jan 23 16:04:52 2004
Thanks for the replies. I now have much reading to do, but I feel I will have the knowledge at the end to make successful samples.
There were a couple of questions on the system I'm looking at. The samples are cobalt oxide deposited onto a lanthanum oxide substrate.
Once I get into the actual hands on portion of sample prep I may be back for some pointers.
Thanks for the swift and copious relies.
Tom
------------------------------------------------------------------------ --------------------------------------------- Thomas M Murray email: murraytm-at-u.washington.edu Electron Microscopy Center manager Phone: (206)543-2836 Materials Science & Engineering Fax: (206)543-3100 Box 352120 302 Roberts Hall Cell: (425)345-0083 University of Washington Home: (425)742-6819 Seattle, WA 98195
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 23 16:36:41 2004
Dear Bill, I have maintained both the Coates and Welter and Zeiss (LEO) DSM982 Gemini FESEMs in this lab. Although a frequent bake out routine is performed, after several thousand hours of operation (~2 years), my SEM gun ion pumps sometimes began to 'burp' intermittently. Without going into all of the possible reasons I guess that most likely the pumps are growing titanium whiskers. Some of those whiskers are shorting out the pump power supply for a fraction of a second until they burn away. Of course the vacuum appears very bad for a split second during that short circuit and the gun power supply kicks off (just when I'm about to leave on vacation). For about 24 years the fix that works for me is to whack the ion pump with the palm of my hand several times before and after a bake (cooled first). (NEVER DO THIS WITH THE FEG ON OR ION PUMP HOT). This causes the vacuum readout to jump several orders of magnitude (into the poorer direction) but after about 30 minutes, the vacuum recovers and remains stable. [I usually whack until the readout begins to drop toward recovery (~2-3 seconds) and a second try for good measure). Firm but cautious taps is the best way I can describe the force of these 'whacks'.]
Caution: If your ion pumps are very used, whacking them may cause titanium flakes to come loose and short the grids permanently. I have been lucky and not lost a pump yet. Depending on the tightness of the vacuum system my ion pumps have worked very well for about 6-10 years each. Before computer interfaces I use to 'hi pot' an ion pump by sending the output of a high voltage spark generator into it for a few seconds (FE emitter grounded to anode, SEM ion pump power supply disconnected and all circuits off and disconnected from the column). That is another way to get rid of whiskers but not recommended as it obviously risks damage to the microscope computer control circuits (and the technician).
Good luck, Jim
Disclaimer: I am not responsible for any damage incurred to you or your equipment if the above methods are used. I am not familiar with an F30H microscope and the vibrations from this method could damage or loosen the emitter or other components in the area of the ion pump. Shock hazard is always present when working with these devices. Refer to qualified personal only.
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James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 3242 91 North Eagleville Road BSP Building, Room G06 Storrs, CT 06269-3242
I assume that you are planning to use gold-labeled antibodies for your experiment (our product, Nanogold, can be used tolabel other types of probes of different sizes; if you are planning to Nanogold-label a much smaller or larger protein and use this to stain your preparation, or if you plan to use a Nanogold labeling reagent to mark a specific chemical group in your specimen, it will likely make a difference to your staining procedure)...in any case, labeling procedures similar to those used for pre-embedding labeling should work well. We have an article on our web site that gives some procedures:
http://www.nanoprobes.com/App3.html
References:
(1) Tanner, V. A., Ploug, T., and Tao-Cheng, J.-H. Subcellular localization of SV2 and other secretory vesicle components in PC12 cells by an efficient method of preembedding EM Immunocytochemistry for cell cultures. J. Histochem. Cytochem., 44, 1481-1488 (1996). (2) Du, J.; Tao-Cheng, J.-H.; Zerfas, P., and McBain, C. J. The K+ channel, Kv2.1, is apposed to astrocytic processes and is associated with inhibitory postsynaptic membranes in hippocampal and cortical principal neurons and inhibitory interneurons. Neuroscience, 84, 37-48 (1998).
If you mean the Nanoprobes product, Nanogold, then we also maintain a comprehensive list of references sorted by application, which includes a section on pre-embedding labeling:
http://www.nanoprobes.com/RefTopNG.html#Npre
Lin et al's paper may also be helpful (Lin, M., Sistina, Y., and Rodger, J. C.: Electron-microscopic localisation of thiol and disulphide groups by direct monomaleimido-Nanogold labeling in the spermatozoa of a marsupial, the tammar wallaby (Macropus eugenii); Cell Tisue Res., 282, 291-296 (1995)).
If negative staining is an option, we offer NanoVan, a negative stain based on vanadium; since vanadium has lower Z than uranium, NanoVan gives a lighter stain that uranyl acetate and allows easier visualization of small gold particles, and can be mixed with our tungsten-based negative stain, Nano-W, to increase density (http://www.nanoprobes.com/Nstain.html). The vast majority of publications by our users describe the use of osmium tetroxide, most frequently after silver enhancement of the gold. A few papers describe the use of uranyl acetate withour osmium tetroxide. Hebert et al used postfixation in 1% osmium tetroxide, 1% potassium ferrocyanide (Hebert, S. S.; Daviau, A.; Grondin, G.; Latreille, M.; Aubin, R. A.; and Blouin, R.: The mixed lineage kinase DLK is oligomerized by tissue transglutaminase during apoptosis. J. Biol. Chem., 275, 32482-90 (2000)).
Hope this helps,
Rick Powell Nanoprobes, Inc.
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
I have a question regarding morphometric analysis, specifically how many individual measurements should be done for a cardiac muscle research project?
The investigators have 4 groups of muscle tissue. Group one, the control has 3 specimens, experimental groups contain 5, 6 & 6 specimens respectively. So that equals 20 specimens total.
I have been asked to measure the width of 20 muscle fibers for each specimen and the diameter of 20 individual fibers on cross section in each group. That adds up to 800 measurements! This seems like overkill. Wouldn't ten do? Does anyone have a reference for how one would measure and what numbers of samples are necessary to provide a statistically appropriate sampling when using a transmission electron microscope.?
Thanks for any advice!
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 Jan 26 03:16:08 2004
I don't have any experience with bromide material, but I will just make you aware that most halides are very difficult to work with in TEM because the specimens are very rapidly filled with radiation-induced defects unless they are cooled to very low temperatures.
-----Original Message----- } From: by way of MicroscopyListserver [mailto:petra.rosner-at-ww.uni-erlangen.de] Sent: 23. januar 2004 15:35 To: microscopy-at-ns.microscopy.com
Dear all in Western/Central Europe, Are there likely to be any courses/workshops on specimen preparation in the near future in western or central Europe? In particular it would be lovely for my colleagues if they are held in German somewhere in one of the German speaking countries. Alternatively, they could be held in English in somewhere reasonably easily reached from the western side of Germany. I can train people to some extent in specimen prep, but it would be nice if they could learn further through courses, hands-on training or such like.
Best wishes
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 26 06:32:31 2004
Your question is a little like "how long is a piece of string"! The number of readings you need will depend mostly on the variation within and between the different samples.
You could try a test within your control group - take the readings and plot a cumulative mean. Eventually the curve will become 'flat' indicating that you have reached a point where there is 'no point' in continuing - the statistical equivalent of empty magnification.
I am a little confused about your use of the words 'width' and 'diameter' - in a cylindrical object, are they not the same thing?
All the best
Gareth Med vänliga hälsningar/With best regards
Gareth
***Come to Stockholm in 2004 to the 26th IFBLS congress*** Take a look at http://www.vardforbundet.se/ifbls2004
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 Mon Jan 26 08:40:58 2004
I don't agree this is overkill. I have done this type of quantitation before, and ended up doing close to 100 measurements per sample! Since these measurements can be done at low magnification, you should be able to measure several fibers from each micrograph, therefore greatly reducing the number of micrographs you need to take. 800 measurements might seem like a lot, but in fact it wouldn't take that long if you are using an appropriate sampling and measurement technique. The fact is you need to sample a lot because of the huge variations you are going to observe, especially if you are measuring fiber width from longitudinal sections as in this case you have no way of knowing if the plane of section is going through the center of the fiber or not. Even when measuring diameters from cross-sections, you expect some variation as sections through the ends of fibers are likely to be smaller than those through the center. Good luck!
Marc
On Sunday, January 25, 2004, at 01:56 PM, Jensen, Karen wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Hello Listers: } } I have a question regarding morphometric analysis, specifically how } many } individual measurements should be done for a cardiac muscle research } project? } } The investigators have 4 groups of muscle tissue. Group one, the } control } has 3 specimens, experimental groups contain 5, 6 & 6 specimens } respectively. So that equals 20 specimens total. } } I have been asked to measure the width of 20 muscle fibers for each } specimen } and the diameter of 20 individual fibers on cross section in each } group. } That adds up to 800 measurements! This seems like overkill. Wouldn't } ten } do? Does anyone have a reference for how one would measure and what } numbers } of samples are necessary to provide a statistically appropriate } sampling } when using a transmission electron microscope.? } } Thanks for any advice! } } 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 } } }
-- 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 Jan 26 09:26:21 2004
Have a look at "Unbiased Stereology" by C.V Howard and M.G. Reed, BIOS Scientific Publishers in the UK, Springer in the USA, 1998. In order to determine if a sample size is "big enough", you will have to do the trial measuerements suggested by Gareth Morgan. Too large a sample wastes your time, too small risks having the paper retuned for more data at a much later time (when details and technique have been forgotten). In my experience measuring things always sounds like more work than it turns out to be. Once you get rolling it is not that bad.
Geoff
Jensen, Karen wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 26 09:59:15 2004
Your question is fairly basic, and that is good, because if it was much more than that, it would be beyond me. {G}
Similar to what Gareth wrote, the measured mean value will stabilize with increased measurements. After multiple measurements, you will be able to determine a mean and standard variation for the population. Both values SHOULD be stable no matter how many measurements you take, but of course it isn't - you have only taken a finite sampling of the entire population. But for multiple random samplings of a population, you will find that the standard deviation in the multiple mean values is equal to the standard deviation of the population divided by the square root of the number of measurements in each sampling.
So consider this example. You take 16 measurements and calculate a mean of 40 um with a standard deviation of 4 um. That 4 um refers to the deviation in any one measurement in the population. How good is your average measurement? It's standard deviation would be 1 um. But suppose you have a different population for which 16 measurements gives you a mean value of 41 also with a standard deviation of 4 um. Are the two populations significantly different? The means are only one standard deviation apart; it is not very certain. But suppose you took 64 measurements per population and obtained the same means and standard deviations. Now the expected deviation in the mean is now 0.5 um due to the increased measurements and the means are now 2 standard deviations apart. That increases the certainty markedly. I haven't dug out my stat tables to verify the exact numbers, but I think the chance of the populations having the same mean dropped from around 37% to around 5% due to the larger sampling. I believe it is the T-test that is used to determine the certainty of such differences.
In your case, cutting the number of measurements from 20 to 10 will lead to a 41% increase in the standard deviation of the mean (SD-10 = SD/Sqrt(10) and SD-20 = SD/Sqrt(20), so SD-10 = Sqrt(2)*SD-20). That may or may not be allowed for the magnitude of the differences between your groups. Indeed, I don't believe it is possible to determine the required sample size without some knowledge of the standard deviations of the populations. Once that is known, you can theoretically set the number of measurements to achieve the desired level of precision, no matter how small; however, you will be constrained by reality. For example, if SD=0.5 units and you wish to detect differences of 0.02 units, you probably want the standard deviation of the mean to be 0.01 units. But that means 0.01=0.5/Sqrt(N), so N=2500 measurements. That is probably not practical. You would probably pick a realistic number of measurements then calculate and report your minimum detectable difference for those conditions. Since you have to quadruple the number of measurements to improve the precision by a factor of 2, you quickly run into practical limits.
It is also probably worth a trip to a statistics book or a stat consultant at that stage. Even as I write, I am remembering principles such as the standard deviation in the difference of means of two populations is the root of the sum of the squares of the deviations of the means of the populations involved. You are testing to see if |Mean(A)-Mean(B)| } 0. It may not be possible to explain such issues succinctly and concisely through e-mail. A book or consultant would probably do better, but I hope this helps some.
Warren
At 12:56 PM 1/25/2004, you wrote:
} Hello Listers: } } I have a question regarding morphometric analysis, specifically how many } individual measurements should be done for a cardiac muscle research } project? } } The investigators have 4 groups of muscle tissue. Group one, the control } has 3 specimens, experimental groups contain 5, 6 & 6 specimens } respectively. So that equals 20 specimens total. } } I have been asked to measure the width of 20 muscle fibers for each specimen } and the diameter of 20 individual fibers on cross section in each group. } That adds up to 800 measurements! This seems like overkill. Wouldn't ten } do? Does anyone have a reference for how one would measure and what numbers } of samples are necessary to provide a statistically appropriate sampling } when using a transmission electron microscope.? } } Thanks for any advice! } } 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 Jan 26 10:12:40 2004
I have found a home for the RCA manuals and parts. I'll be responding individually to everyone who answered my query shortly. Thanks to all.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 26 10:24:39 2004
South Bay Technology regularly conducts specimen preparation workshops around the world as well as in our own facility here in San Clemente, CA. We also do cutomized workshops for customers that we can do at your facility. We cover any topic in specimen preparation: Tripod Polishing, Dimpling, Low Energy Ion Milling, Jet Polishing, Plasma Cleaning, Plasma Trimming, Pre-FIB preparation, MicroCLeaving etc. Please contact me off-line and I will let you know what we have planned and what we can arrange to meet your needs.
Best regards-
David
Ian MacLaren 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 all in Western/Central Europe, } Are there likely to be any courses/workshops on specimen preparation in } the near future in western or central Europe? In particular it would be } lovely for my colleagues if they are held in German somewhere in one of } the German speaking countries. Alternatively, they could be held in } English in somewhere reasonably easily reached from the western side of } Germany. } I can train people to some extent in specimen prep, but it would be nice } if they could learn further through courses, hands-on training or such } like. } } Best wishes } } -- } Ian MacLaren } Technische Universit?t Darmstadt } Material- und Geowissenschaften } Petersenstr. 23 } 64287 Darmstadt } Germany } http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 26 13:28:25 2004
Looking for opinions, comments, pros & cons, etc. in comparing a Low Vacuum-SEM with a tungsten gun vs a Lab6 gun.
Details: One of the primary applications presently is EBSD and XEDS of geologic samples, of these one of the primary sample types is very fine (nano) crystals ranging 10-100nm in size. And it is for these types of samples we are strongly considering LAB6. Our thoughts are smaller spot size and increased beam current, which will improve our diffraction signal, as well improving our ability to locate the particles. Secondly, the scope will primarily be operated by neophyte SEM users, users who are primarily concerned with generating good EBSD and XEDS data not SEI/BEI imaging (yeah, yeah but I don’t wish to open that discussion again).
So I am seeking comments: are the advantages of LAB6 real under these conditions? How do these LAB6 advantages compare to the costs of purchase and operation? And would a better recommendation be to spend funds for other items like an “after-market” BSE detector (i.e. Robinson, etc.).
Thank you!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 26 14:03:42 2004
I need some tips on EBSP sample prepration for a Zn alloy with additions on Bi and In that have concentrations of {0.3 wt.%. I wish to characterize these phases, which are {1 micron in size. TEM has not worked for me. The material is in powder form, but I have managed to compact them into disks for handling. I tried electropolishing with phosphoric acid and ethanol, and got good results for the Zn matrix (even got a Kikuchi pattern), but I was unable to get a pattern for the Bi-In phases. I believe that these are tarnishing. Can somebody recommend an EBSP sample prep method using mechanical polishing or ion etch? I have a Buehler Vibromet and a PIP tool that may help me. I am still open to electropolishing if anybody has any ideas. All prepared samples will be placed on an FESEM. All responses are greatly appreciated. Thank you! :o)
Martin Perez Ph D student University of Missouri-Rolla
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 26 14:23:48 2004
Yes, folks I realize that across the board microscope vendors have moved to concentrate on both W- or FEG scopes, with LaB6 left by the way side or in the middle. And yes, FEG is the way to go - but with two caveats: (1) FEG is $120-190K more than W- for every thing else the same, and (2) service contracts are $17-20K more per year. So these are two BIG reasons to look at LaB6.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 26 15:11:21 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Coming from a physical sciences background, this is something that has often puzzled me - apologies if I have missed something obvious.
When making measurements on regular biological structures, such as muscle fibres, why not use diffraction?
The big advantage, it would seem to me, is that a single diffraction pattern immediately and automatically gives average spacings over a large number of individual fibres, something that would require several images and a great deal of tedious measurement (almost guaranteed to lead to errors).
In this case, couldn't 20 diffraction patterns (one from each sample) give statistically better data than the 800 measurements from images?
Of course, there are disadvantages - you need a TEM capable of operating at long camera lengths (although most, as far as I am aware, can) and you need to calibrate the TEM for operation at such camera lengths. Even so, I still think you would take less time and the results would be statistically better. -- Larry Stoter
NOTE - any message other than plain text will be automatically deleted :-)
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 26 18:27:36 2004
Most of the previous responders have made valid points, but many important issues have been neglected. First you stated
"I have been asked to measure the width of 20 muscle fibers for each specimen and the diameter of 20 individual fibers on cross section in each group."
Do you mean that you need to measure the diameter of individual myofibrils in addition to the diameter of the myofibers? And how do you intend to measure the "diameter" or "width"? of these structures? Traditionally, muscle fibers (in cross section) are measured by their "least diameter", which can be defined as the distance between the closest parallel lines that can be drawn around the fiber. This measurement is used because it is less sensitive to error caused by poor orientation than other values, such as area, circumference, or maximum diameter.
It will be much quicker and cost effective to measure the myofibers by light microscopy. If myofibrils must also be measured, TEM will obviously be required for those measurements. But for both types of measurements, careful thought must be given to the sampling procedure as well as the total number of structures measured. Because muscle fibers are more like irregular polyhedrons than cylinders, even the least diameter statistic is affected to some extent by orientation. Heart tissue is particularly problematic because the fibers are not all parallel, and fascicles will be seen in different orientation in the same section. When a muscle fiber contracts, it's diameter increases. During fixation of muscle it is common for some areas to become hyper contracted and other areas to become hyper extended. Skeletal muscle is "stretched" during fixation with some kind of clamping device to minimize this, but this is rarely done with heart. The diameters of the fibers and fibrils will also be affected by the quality of the fixation and the osmolarity of the fixative. You may well see variation between areas on the surface of the sample versus those deeper in.
The upshot of this is that even if large numbers of fibers are measured from each sample, the results will be meaningless unless the sampling procedure is appropriate. There may well be greater variations between areas within each sample than between the samples as a whole. You may need to do an analysis of variance (ANOVA) looking at variation within an area of a sample, between different areas of a sample, between samples with the same treatment, and between treatments, in order to determine if there is a real difference between the treatments. If you are trying to find subtle differences, 800 measurements may not be nearly enough. Also, your investigators may be interested in the distribution of fiber diameters (the results are usually displayed as histograms), not just the means, and that would require even larger numbers. If the samples are not well prepared and handled identically, the project may be hopeless.
The good news is that the least diameter measurements are not difficult or time consuming if you have an appropriate morphometry program. I routinely measure 400-800 fibers per case for muscle biopsies, and that takes less than 2 hours. If you do not have a morphometry package, NIH Image (for Mac) and SCION Image (for Windows) are available as free downloads.
Ralph Common Electron Microscopist Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824 517-355-7558; fax 517-432-1053 ralph.common-at-ht.msu.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 26 22:49:34 2004
I have two separate issues regarding 16-bit TIFF files that I cannot get resolved. Perhaps someone has run into one or both of these issues.
EDAX: The direct active image capture will go up to 4096 or in my case, 8192 pixels at 1.33 aspect ratio. However, the images are 8-bit. Well, the image capture ADC is 12-bits...4096 shades. There does not seem a way to capture an image as 16-bit TIFF. Very frustrating. Is there some way around this?
LEO: The LEO32 GUI will save TIFF files as 16-bit but the files are not readable on any system other than the LEO SEM. The 8-bit files are standard TIFF. The LEO ADC is also 12-bit. So why can't the 16-bit TIFF be read by some other app? Photoshop will not read it. ImageJ will not read it. Nothing I have will.
Any ideas?
tnx, gary g.
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 00:00:13 2004
If you are compressing the powders in to a pellet, there is a high risk of plastically deforming phases enough to blur any pattern. In that case, no prep technique will yield patterns.
John Mardinly 408-765-2346 877-277-1182
-----Original Message----- } From: Perez, Martin Gerardo (UMR-Student) [mailto:mperez-at-umr.edu] Sent: Monday, January 26, 2004 12:15 PM To: Microscopy-at-MSA.Microscopy.Com
Dear MSA members,
I need some tips on EBSP sample prepration for a Zn alloy with additions on Bi and In that have concentrations of {0.3 wt.%. I wish to characterize these phases, which are {1 micron in size. TEM has not worked for me. The material is in powder form, but I have managed to compact them into disks for handling. I tried electropolishing with phosphoric acid and ethanol, and got good results for the Zn matrix (even got a Kikuchi pattern), but I was unable to get a pattern for the Bi-In phases. I believe that these are tarnishing. Can somebody recommend an EBSP sample prep method using mechanical polishing or ion etch? I have a Buehler Vibromet and a PIP tool that may help me. I am still open to electropolishing if anybody has any ideas. All prepared samples will be placed on an FESEM. All responses are greatly appreciated. Thank you! :o)
Martin Perez Ph D student University of Missouri-Rolla
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 02:18:25 2004
*Subject: [Microscopy] Need help: EBSP sample prep *Date sent: Mon, 26 Jan 2004 14:14:40 -0600 *From: "Perez, Martin Gerardo (UMR-Student)" {mperez-at-umr.edu} *To: {Microscopy-at-MSA.Microscopy.Com}
* * *------------------------------------------------------------------------------ *The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello:
Maybe it would be goog idea to check www.hkltechnology.com page. There is a lot of useful information and papers too.
Good luck.
Best regards,
Witold Zielinski
*Dear MSA members, * *I need some tips on EBSP sample prepration for a Zn alloy with additions on Bi and In that have concentrations of {0.3 wt.%. I wish to characterize these phases, which are {1 micron in size. TEM ha
* not worked for me. The material is in powder form, but I have managed to compact them into disks for handling. I tried electropolishing with phosphoric acid and ethanol, and got good results for
*he Zn matrix (even got a Kikuchi pattern), but I was unable to get a pattern for the Bi-In phases. I believe that these are tarnishing. Can somebody recommend an EBSP sample prep method using mech
*nical polishing or ion etch? I have a Buehler Vibromet and a PIP tool that may help me. I am still open to electropolishing if anybody has any ideas. All prepared samples will be placed on an FESE
*. All responses are greatly appreciated. Thank you! :o) * *Martin Perez *Ph D student *University of Missouri-Rolla * * * :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) ;)
Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
Back in sand free civilisation again after my trip to Egypt I note that not one person took up my challenge to take a look at their laboratories performance by way of www.iaaem.com/mppa1 .
We all seem to agree that the standards of EM output that we see in journals have dropped. You may not agree with what the quality people have developed, but you do have to admit that we do not have, anywhere in the world, a standard by which EM units may be judged. Country by country the ISO/Quality procedures are moving in. In time everyone will have to conform to some form of quality standard, why not get ahead of the game?
In Australia the Quality movement is being spread country wide fast and furious. Building on their theme of inter laboratory co-operation on instrumentation they seem to be tackling the future of electron microscopy with a very positive attitude Not all will survive, so they are making sure everything is being done working together to set a very firm foundation for their future.
On my travels it frightens me when I see how good some laboratories are and how poor are some others. It is not the actual standard that really concerns me, it is the perceived position of these laboratories within a specific region. Are the perceived top laboratories really putting out the best work, or do they just rely upon their reputation?
I hope this is worth a thought?
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 04:56:45 2004
We got used Zeiss CEM902. After installation and initial switch on a resistor in the Lens Current Unit(?) and a wired connection between resistor and condenser in the Power Supply were burned out. To find out the reason(s) we need Circuit Diagrams. I would be very pleased if anyone could send theses about appropriate units.
Dr.Raivo Raid, PhD
Institute of Zoology&Hydrobiology University of Tartu Vanemuise 46 51014 Tartu Estonia Tel. +372 7 375840 Fax +372 7 375830
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 07:55:46 2004
It might be worthwhile to take a look at the following papers from a group at the University in Saarbrucken in Germany. Both papers provide extensive information on the optimal polishing of a wide range of materials.
1. Katrakova, D., Mücklich, F.(2001) Specimen Preparation for Electron Backscatter Diffraction - Part I: Metals Prakt. Metallogr. 38, 10, pp.547-565
2. Katrakova, D., Mücklich, F.(2001) Specimen Preparation for Electron Backscatter Diffraction - Part II: Ceramics Prakt. Metallogr. 39, 12, pp.644-662
Rene de Kloe EDAX-TSL
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, 27 January, 2004 07:11 To: Perez, Martin Gerardo (UMR-Student); Microscopy-at-MSA.Microscopy.Com
Email: ecd10-at-psu.edu Name: Prof. Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Microscopy] [Filtered] MListserver: TEM Facility Manager Opening
Question: The Materials Research Institute at the Pennsylvania State University invites applications for a Transmission Electron Microscopist to oversee its TEM facilities. The facility supports a wide variety of materials research programs at Penn State. The Facility Manager will have primary responsibility for management and operation of the JEOL 2010F, JEOL 2010 LaB6 and Philips CM20 TEMs and auxiliary equipment, which reside in the facility. The Facility Manager will oversee training of PSU students, faculty and staff on TEM-related equipment and software and assist users with performing advanced microscopy techniques.
A MS is required with a PhD preferred in Materials, Physical Sciences or Engineering. The prospective candidate must have a strong background in the theory and practice of TEM microcharacterizaton with a minimum of 5 years post-graduate experience. The candidate should also have experience with vacuum technology, electronics, and computers. Strong interpersonal and communications skills are essential.
Interested candidates should send a resume and cover letter to Prof. Elizabeth Dickey, The Pennsylvania State University, 223 MRL Building, University Park, PA 16802. In addition, candidates should arrange to have three letters of recommendation sent to the above address.
Penn State is committed to affirmative action, equal opportunity and the diversity of its workforce.
Does the TIFF images that are saved in 16 bit format have a "TIF" extension or are they similar to a "composite" image as would be saved by PCI Quartz for example? I ask because PCI Quartz saves both the TIF image and the data that has annotations on it and can only be opened in that form by the PCI application.
Peter Tomic Agere Systems
Peter -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Tuesday, January 27, 2004 12:01 AM To: MSA listserver
Hello all:
I have two separate issues regarding 16-bit TIFF files that I cannot get resolved. Perhaps someone has run into one or both of these issues.
EDAX: The direct active image capture will go up to 4096 or in my case, 8192 pixels at 1.33 aspect ratio. However, the images are 8-bit. Well, the image capture ADC is 12-bits...4096 shades. There does not seem a way to capture an image as 16-bit TIFF. Very frustrating. Is there some way around this?
LEO: The LEO32 GUI will save TIFF files as 16-bit but the files are not readable on any system other than the LEO SEM. The 8-bit files are standard TIFF. The LEO ADC is also 12-bit. So why can't the 16-bit TIFF be read by some other app? Photoshop will not read it. ImageJ will not read it. Nothing I have will.
Any ideas?
tnx, gary g.
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 09:42:11 2004
} Country by country the ISO/Quality procedures are } moving in. In time everyone will have to conform to some form of quality } standard ...
Yes, but my problem with ISO9000 and its ilk is that it is much more concerned with documentation and reproducibility than with what you and I would call quality. Reproducible mediocrity, generated by a well-defined procedure, is just fine. In fact, I might argue that your perceived decline in average quality is CAUSED by the assumption that as long as it's ISO compliant, it's good enough, so there's no point in investing additional time to optimize. Cut-and-dried canned procedures allow managers to use lower-level personnel.
Additionally, IMO, the documentation burden inhibits innovation by encasing whatever you do now in concrete. That is certainly true for small US manufacturers in conforming to CE requirements. The cost of change is insignificant for a large consumer electronics firm, but quite large for the smaller companies that largely serve the microscopy field.
And who's to say that's not what the majority of the customer base actually wants? Predictability over excellence or innovation?
Rick Mott
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 09:45:44 2004
I have a Durst Enlarger with all accessories and a Ilford light source now. I have a interested party, but have no idea as to what or how to price the instrument?
Any suggestions would be helpful..
Eric UCLA Medical Center Electron Microscopy Lab
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 11:01:39 2004
These are as they came off of a Supra 55. They are named tif by the system. Windows TIFF reader does not like the 8-bit files but Photoshop does. The 16-bit files come out as grey snow--all garbaged bits. ImageJ correctly reads the image frame size (1024x768) and has a offset but it too can't read the image file.
For 16-bit analySIS files, save as 16-bits and do Image/Auto Level in Photoshop.
gary g.
At 07:18 AM 1/27/2004, you wrote: } Gary; } } Does the TIFF images that are saved in 16 bit format have a "TIF" } extension or are they similar to a "composite" image as would be saved by } PCI Quartz for example? I ask because PCI Quartz saves both the TIF image } and the data that has annotations on it and can only be opened in that } form by the PCI application. } } Peter Tomic } Agere Systems } } Peter } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Tuesday, January 27, 2004 12:01 AM } To: MSA listserver } Subject: [Microscopy] EDAX and LEO 16-bit files } } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 13:07:28 2004
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Martin,
It might be worthwhile to take a look at the following papers from a group at the University in Saarbrucken in Germany. Both papers provide extensive information on the optimal polishing of a wide range of materials.
1. Katrakova, D., Mücklich, F.(2001) Specimen Preparation for Electron Backscatter Diffraction - Part I: Metals Prakt. Metallogr. 38, 10, pp.547-565
2. Katrakova, D., Mücklich, F.(2001) Specimen Preparation for Electron Backscatter Diffraction - Part II: Ceramics Prakt. Metallogr. 39, 12, pp.644-662
Rene de Kloe EDAX-TSL
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, 27 January, 2004 07:11 To: Perez, Martin Gerardo (UMR-Student); Microscopy-at-MSA.Microscopy.Com
Gary:
The 16 bit image you posted does open correctly in Graphics Converter if you tell it to swap the order of the bytes. When opened in Photoshop, what you are apparently seeing is the low byte (essentially random values) first. That suggests that the Leo folks set one of the (many) flags in the tif format wrong. Tif is really a collection of possibilities rather than a well defined format. There are so many combinations that not even Adobe, who is the caretaker of the standard, opens all of the legal variants in programs like Photoshop. I've seen someone's estimate that there are at least 160 combinations of legal settings. If in addition to that, someone gets one of the flags wrong, there is no limit to the confusion that can result. this time it looks like the fault is Leo's but in general many companies use and abuse the tif format. Still, it's better than something that is purely proprietary and undocumented.
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 13:18:15 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Email: ecd10-at-psu.edu Name: Prof. Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Microscopy] [Filtered] MListserver: TEM Facility Manager Opening
Question: The Materials Research Institute at the Pennsylvania State University invites applications for a Transmission Electron Microscopist to oversee its TEM facilities. The facility supports a wide variety of materials research programs at Penn State. The Facility Manager will have primary responsibility for management and operation of the JEOL 2010F, JEOL 2010 LaB6 and Philips CM20 TEMs and auxiliary equipment, which reside in the facility. The Facility Manager will oversee training of PSU students, faculty and staff on TEM-related equipment and software and assist users with performing advanced microscopy techniques.
A MS is required with a PhD preferred in Materials, Physical Sciences or Engineering. The prospective candidate must have a strong background in the theory and practice of TEM microcharacterizaton with a minimum of 5 years post-graduate experience. The candidate should also have experience with vacuum technology, electronics, and computers. Strong interpersonal and communications skills are essential.
Interested candidates should send a resume and cover letter to Prof. Elizabeth Dickey, The Pennsylvania State University, 223 MRL Building, University Park, PA 16802. In addition, candidates should arrange to have three letters of recommendation sent to the above address.
Penn State is committed to affirmative action, equal opportunity and the diversity of its workforce.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Email: ecd10-at-psu.edu Name: Prof. Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Microscopy] [Filtered] MListserver: TEM Facility Manager Opening
Question: The Materials Research Institute at the Pennsylvania State University invites applications for a Transmission Electron Microscopist to oversee its TEM facilities. The facility supports a wide variety of materials research programs at Penn State. The Facility Manager will have primary responsibility for management and operation of the JEOL 2010F, JEOL 2010 LaB6 and Philips CM20 TEMs and auxiliary equipment, which reside in the facility. The Facility Manager will oversee training of PSU students, faculty and staff on TEM-related equipment and software and assist users with performing advanced microscopy techniques.
A MS is required with a PhD preferred in Materials, Physical Sciences or Engineering. The prospective candidate must have a strong background in the theory and practice of TEM microcharacterizaton with a minimum of 5 years post-graduate experience. The candidate should also have experience with vacuum technology, electronics, and computers. Strong interpersonal and communications skills are essential.
Interested candidates should send a resume and cover letter to Prof. Elizabeth Dickey, The Pennsylvania State University, 223 MRL Building, University Park, PA 16802. In addition, candidates should arrange to have three letters of recommendation sent to the above address.
Penn State is committed to affirmative action, equal opportunity and the diversity of its workforce.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gary;
Does the TIFF images that are saved in 16 bit format have a "TIF" extension or are they similar to a "composite" image as would be saved by PCI Quartz for example? I ask because PCI Quartz saves both the TIF image and the data that has annotations on it and can only be opened in that form by the PCI application.
Peter Tomic Agere Systems
Peter -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Tuesday, January 27, 2004 12:01 AM To: MSA listserver
Hello all:
I have two separate issues regarding 16-bit TIFF files that I cannot get resolved. Perhaps someone has run into one or both of these issues.
EDAX: The direct active image capture will go up to 4096 or in my case, 8192 pixels at 1.33 aspect ratio. However, the images are 8-bit. Well, the image capture ADC is 12-bits...4096 shades. There does not seem a way to capture an image as 16-bit TIFF. Very frustrating. Is there some way around this?
LEO: The LEO32 GUI will save TIFF files as 16-bit but the files are not readable on any system other than the LEO SEM. The 8-bit files are standard TIFF. The LEO ADC is also 12-bit. So why can't the 16-bit TIFF be read by some other app? Photoshop will not read it. ImageJ will not read it. Nothing I have will.
Any ideas?
tnx, gary g.
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 15:09:10 2004
Question: I am looking for a firm to service the Los Angeles County Museum's Cambridge Stereoscan 200 SEM. We are looking for a someone to assess the health of the instrument, to provide recommendations for upgrades, and then to possibly provide ongoing emergency service. Could anyone recommend such a company?
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Listers:
We got used Zeiss CEM902. After installation and initial switch on a resistor in the Lens Current Unit(?) and a wired connection between resistor and condenser in the Power Supply were burned out. To find out the reason(s) we need Circuit Diagrams. I would be very pleased if anyone could send theses about appropriate units.
Dr.Raivo Raid, PhD
Institute of Zoology&Hydrobiology University of Tartu Vanemuise 46 51014 Tartu Estonia Tel. +372 7 375840 Fax +372 7 375830
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 15:59:17 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Steve Chapman wrote:
} Country by country the ISO/Quality procedures are } moving in. In time everyone will have to conform to some form of quality } standard ...
Yes, but my problem with ISO9000 and its ilk is that it is much more concerned with documentation and reproducibility than with what you and I would call quality. Reproducible mediocrity, generated by a well-defined procedure, is just fine. In fact, I might argue that your perceived decline in average quality is CAUSED by the assumption that as long as it's ISO compliant, it's good enough, so there's no point in investing additional time to optimize. Cut-and-dried canned procedures allow managers to use lower-level personnel.
Additionally, IMO, the documentation burden inhibits innovation by encasing whatever you do now in concrete. That is certainly true for small US manufacturers in conforming to CE requirements. The cost of change is insignificant for a large consumer electronics firm, but quite large for the smaller companies that largely serve the microscopy field.
And who's to say that's not what the majority of the customer base actually wants? Predictability over excellence or innovation?
Rick Mott
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 27 16:32:58 2004
I have used horseradish peroxidase as a tracer in vivo in the past and am aware that it can damage cells and cross membranes with prolonged exposure. I am now trying to find a reference that demonstrates this fact. A literature search using Medline is difficult on this topic since it is hard to come up with a good Boolean logic search of keywords (this is one of two problems that I am posting that I am having a difficult time searching the literature for). Any citations would be greatly appreciated. TIA, tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I know this is a longshot but has anyone ever seen a paper where someone looked at a sample by SEM and then embedded the sample and looked at the exact same cells by TEM? A literature search using Medline is difficult on this topic since it is hard to come up with a good Boolean logic search of keywords that doesn't pull up 1000's of non-relevant TEM & SEM papers. (this is one of two problems that I am posting that I am having a difficult time searching the literature for). Any citations would be greatly appreciated. TIA, tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I did quite a bit of this sort of work many years ago. Works quite well although the organelle ultrastructure does suffer a little. Don't expect the same clarity of membranes as a specimen processed directly for TEM.
The text 'Principles and Techniques of Scanning Electron Microscopy' by Hayat, has a chapter outlining the method. Chapter 5 written by M.Gary Wickham and David M. Worthen.
Also Chaptr 11 written by Willis K. Paul.
In our library this text had the code QH 212.53. If you wish I could check whether our library still has this book for an ISBN number. We are talking 1974 edition.
Other references at the time,
Anat Rec (1973) Vol 176, pp 245-252. Samuel M. Meller, et al. Transmission electron microscopy of critical point dried tissue after observation in the scanning electron microscope.
Cell Tissue Research Vol 162 pp 61-73 (1975) D.E. Scott, et al. The Primate Median Eminence. Correlative scanning transmission electron microscopy.
Cell Tissue Research Vol 190 pp 317-336 (1978) D.E. Scott, et al. Coorelative Scanning-Transmission EM Examination of the Perinatal rat brain.
Human Reproduction (1991), Vol 6 pp 645-660, and Human Reproduction (1992), Vol 7, pp 446 - 452. W.R. Gillet et al, Both to do with human ovary, examined in SEM then same sample examined in TEM.
At the time we played around with doing SEM on blocks that had ultrathins cut from them. Worked really well also.
Hope this gets you started
Allan
} } I know this is a longshot but has anyone ever seen a paper where } someone looked at a sample by SEM and then embedded the sample and } looked at the exact same cells by TEM? A literature search using } Medline is difficult on this topic since it is hard to come up with } a good Boolean logic search of keywords that doesn't pull up 1000's } of non-relevant TEM & SEM papers. (this is one of two problems that } I am posting that I am having a difficult time searching the } literature for). Any citations would be greatly appreciated. TIA, } tom } } Thomas E. Phillips, PhD -- ------------------------------------------------- Allan Mitchell Technical Manager Otago Centre for Electron Microscopy C/-Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
----------------------------------------------------------------- 2005 Microscopy Conference in Dunedin: http://ocem.otago.ac.nz/Conference_2005/Microscopy_NZ_2005.html
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Martin,
It might be worthwhile to take a look at the following papers from a group at the University in Saarbrucken in Germany. Both papers provide extensive information on the optimal polishing of a wide range of materials.
1. Katrakova, D., Mücklich, F.(2001) Specimen Preparation for Electron Backscatter Diffraction - Part I: Metals Prakt. Metallogr. 38, 10, pp.547-565
2. Katrakova, D., Mücklich, F.(2001) Specimen Preparation for Electron Backscatter Diffraction - Part II: Ceramics Prakt. Metallogr. 39, 12, pp.644-662
Rene de Kloe EDAX-TSL
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, 27 January, 2004 07:11 To: Perez, Martin Gerardo (UMR-Student); Microscopy-at-MSA.Microscopy.Com
Search for Gareth Griffiths, he has done a lot of HRP experiments as I know Good luck danièle
-----Message d'origine----- De : Tom Phillips [mailto:phillipst-at-missouri.edu] Envoyé : mardi 27 janvier 2004 23:45 À : Microscopy-at-msa.microscopy.com Objet : HRP damage to membranes in vivo
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I have used horseradish peroxidase as a tracer in vivo in the past and am aware that it can damage cells and cross membranes with prolonged exposure. I am now trying to find a reference that demonstrates this fact. A literature search using Medline is difficult on this topic since it is hard to come up with a good Boolean logic search of keywords (this is one of two problems that I am posting that I am having a difficult time searching the literature for). Any citations would be greatly appreciated. TIA, tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Hi everybody! My University have received some money to buy an ion milling (with a dimple grinder, a disc puncher, a disc grinder etc) to prepare samples for TEM. I have to decide between Gatan and Fischione. I would appreciate very much any opinion about these two companies because I have to make a decision as soon as possible. Thank you very much Cristina Almansa University of Alicante, Spain
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 28 04:38:41 2004
Hi everybody! My University have received some money to buy an ion milling (with a dimple grinder, a disc puncher, a disc grinder etc) to prepare samples for TEM. I have to decide between Gatan and Fischione. I would appreciate very much any opinion about these two companies because I have to make a decision as soon as possible. Thank you very much Cristina Almansa University of Alicante, Spain
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 28 05:35:15 2004
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------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Martin,
It might be worthwhile to take a look at the following papers from a group at the University in Saarbrucken in Germany. Both papers provide extensive information on the optimal polishing of a wide range of materials.
1. Katrakova, D., Mücklich, F.(2001) Specimen Preparation for Electron Backscatter Diffraction - Part I: Metals Prakt. Metallogr. 38, 10, pp.547-565
2. Katrakova, D., Mücklich, F.(2001) Specimen Preparation for Electron Backscatter Diffraction - Part II: Ceramics Prakt. Metallogr. 39, 12, pp.644-662
Rene de Kloe EDAX-TSL
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, 27 January, 2004 07:11 To: Perez, Martin Gerardo (UMR-Student); Microscopy-at-MSA.Microscopy.Com
I sent the image that Gary posted to Chris Cox at Adobe. Adobe is the caretaker of the TIFF standard and Chris is the person who is directly in charge of the Photoshop code that reads and writes files. His reply is below. Somebody needs to tell LEO to correct their file writing routine!
} From: Chris Cox {ccox-at-adobe.com} } Date: January 27, 2004 10:18:56 PM EST } } The file in question is little endian, 16 bit/sample, 1 sample, } uncompressed, black is zero photometric interpretation. } } As far as I can tell, Photoshop is reading the image 100% correctly. } } BUT - the file is written incorrectly. } They handled the byte order on the tags, but failed to handle the byte } order on the image data. } If I change the byte order info halfway through the decode process in } Photoshop, we read the image correctly (SEM image of what looks like a } semiconductor wafer surface showing one trace and one partial trace } plus some pits). } } So, it's up to whomever wrote this file to fix their TIFF writing code. } I can't believe they'd make such a novice mistake.
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 28 06:51:40 2004
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I put the same image as 8-bit and 16-bit here:
www.microtechnics.com/leo_td_08.tif
www.microtechnics.com/leo_td_16.tif
These are as they came off of a Supra 55. They are named tif by the system. Windows TIFF reader does not like the 8-bit files but Photoshop does. The 16-bit files come out as grey snow--all garbaged bits. ImageJ correctly reads the image frame size (1024x768) and has a offset but it too can't read the image file.
For 16-bit analySIS files, save as 16-bits and do Image/Auto Level in Photoshop.
gary g.
At 07:18 AM 1/27/2004, you wrote: } Gary; } } Does the TIFF images that are saved in 16 bit format have a "TIF" } extension or are they similar to a "composite" image as would be saved by } PCI Quartz for example? I ask because PCI Quartz saves both the TIF image } and the data that has annotations on it and can only be opened in that } form by the PCI application. } } Peter Tomic } Agere Systems } } Peter } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Tuesday, January 27, 2004 12:01 AM } To: MSA listserver } Subject: [Microscopy] EDAX and LEO 16-bit files } } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 28 06:53:30 2004
Question: I am looking for a firm to service the Los Angeles County Museum's Cambridge Stereoscan 200 SEM. We are looking for a someone to assess the health of the instrument, to provide recommendations for upgrades, and then to possibly provide ongoing emergency service. Could anyone recommend such a company?
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Email: ecd10-at-psu.edu Name: Prof. Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Microscopy] [Filtered] MListserver: TEM Facility Manager Opening
Question: The Materials Research Institute at the Pennsylvania State University invites applications for a Transmission Electron Microscopist to oversee its TEM facilities. The facility supports a wide variety of materials research programs at Penn State. The Facility Manager will have primary responsibility for management and operation of the JEOL 2010F, JEOL 2010 LaB6 and Philips CM20 TEMs and auxiliary equipment, which reside in the facility. The Facility Manager will oversee training of PSU students, faculty and staff on TEM-related equipment and software and assist users with performing advanced microscopy techniques.
A MS is required with a PhD preferred in Materials, Physical Sciences or Engineering. The prospective candidate must have a strong background in the theory and practice of TEM microcharacterizaton with a minimum of 5 years post-graduate experience. The candidate should also have experience with vacuum technology, electronics, and computers. Strong interpersonal and communications skills are essential.
Interested candidates should send a resume and cover letter to Prof. Elizabeth Dickey, The Pennsylvania State University, 223 MRL Building, University Park, PA 16802. In addition, candidates should arrange to have three letters of recommendation sent to the above address.
Penn State is committed to affirmative action, equal opportunity and the diversity of its workforce.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Email: ecd10-at-psu.edu Name: Prof. Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Microscopy] [Filtered] MListserver: TEM Facility Manager Opening
Question: The Materials Research Institute at the Pennsylvania State University invites applications for a Transmission Electron Microscopist to oversee its TEM facilities. The facility supports a wide variety of materials research programs at Penn State. The Facility Manager will have primary responsibility for management and operation of the JEOL 2010F, JEOL 2010 LaB6 and Philips CM20 TEMs and auxiliary equipment, which reside in the facility. The Facility Manager will oversee training of PSU students, faculty and staff on TEM-related equipment and software and assist users with performing advanced microscopy techniques.
A MS is required with a PhD preferred in Materials, Physical Sciences or Engineering. The prospective candidate must have a strong background in the theory and practice of TEM microcharacterizaton with a minimum of 5 years post-graduate experience. The candidate should also have experience with vacuum technology, electronics, and computers. Strong interpersonal and communications skills are essential.
Interested candidates should send a resume and cover letter to Prof. Elizabeth Dickey, The Pennsylvania State University, 223 MRL Building, University Park, PA 16802. In addition, candidates should arrange to have three letters of recommendation sent to the above address.
Penn State is committed to affirmative action, equal opportunity and the diversity of its workforce.
I have been discussing with various people the the use of the following words
dye stain label marker
The problem has been the use of the word "label". There exists two opinion about the proper use of it. The one states strictly that a label has always to be coupled to the target with antibody-antigen reaction. The other school states that a label is generally a molecule or an atom or a bead etc that makes the target visible in an instrument (eg. a microscope). So plese someone, give me an explanation what exactly is a "label" ?
Thank you.
Regards Ari Kuusisto Research physicist
PerkinElmer Life and Analytical Sciences tel. +358-2-2678 508 fax. +358-2-2578 357 E-mail: Ari.Kuusisto-at-PerkinElmer.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 28 08:30:14 2004
Question: I am looking for a firm to service the Los Angeles County Museum's Cambridge Stereoscan 200 SEM. We are looking for a someone to assess the health of the instrument, to provide recommendations for upgrades, and then to possibly provide ongoing emergency service. Could anyone recommend such a company?
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I sent the image that Gary posted to Chris Cox at Adobe. Adobe is the caretaker of the TIFF standard and Chris is the person who is directly in charge of the Photoshop code that reads and writes files. His reply is below. Somebody needs to tell LEO to correct their file writing routine!
} From: Chris Cox {ccox-at-adobe.com} } Date: January 27, 2004 10:18:56 PM EST } } The file in question is little endian, 16 bit/sample, 1 sample, } uncompressed, black is zero photometric interpretation. } } As far as I can tell, Photoshop is reading the image 100% correctly. } } BUT - the file is written incorrectly. } They handled the byte order on the tags, but failed to handle the byte } order on the image data. } If I change the byte order info halfway through the decode process in } Photoshop, we read the image correctly (SEM image of what looks like a } semiconductor wafer surface showing one trace and one partial trace } plus some pits). } } So, it's up to whomever wrote this file to fix their TIFF writing code. } I can't believe they'd make such a novice mistake.
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 28 12:52:43 2004
} I have been discussing with various people the the use of the following } words } } dye } stain } label } marker } } The problem has been the use of the word "label". There exists two } opinion } about the proper use of it. The one states strictly that a label has } always } to be coupled to the target with antibody-antigen reaction. The other } school } states that a label is generally a molecule or an atom or a bead etc } that } makes the target visible in an instrument (eg. a microscope). So plese } someone, give me an explanation what exactly is a "label" ?
Dear Ari, I would define a dye as a substance that imparts color to a substrate. It need not be specific, so the color could be uniform, such as hair or cloth dyes. A stain is a substance that imparts visibility to a substrate, and in the context of microscopy a stain has some specificity. For LM a stain generally imparts color (i.e., is a type of dye), and for EM a stain is a stronger scatterer of electrons than is the substrate. A marker is a substance that adds a recognizable feature to a substrate. Finally, a label is a marker that is associated with a particular component of the substrate. Thus, there are fluorescent dyes that are lipophyllic, so these can be used as labels for membranes, there are molecules that bind very specifically, including biotin-avidin, so including but not restricted to antibody-antigen pairs. These can be bound to markers, such as green fluorescent protein or colloidal gold, and I call these labels. 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 Jan 28 13:20:24 2004
Yes, I see the duplicate messages I am attempting to sort out why it started it may have to do with the virus attack which is going on in the USA right now.
Unfortunately, I am on the road on the way to the Australian meeting. This may take me some time to fix as my connectivity will be intermittent.
Nestor Your Friendly Neighborhood SysOp.
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 28 13:27:03 2004
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Dear Microscopists,
I have been discussing with various people the the use of the following words
dye stain label marker
The problem has been the use of the word "label". There exists two opinion about the proper use of it. The one states strictly that a label has always to be coupled to the target with antibody-antigen reaction. The other school states that a label is generally a molecule or an atom or a bead etc that makes the target visible in an instrument (eg. a microscope). So plese someone, give me an explanation what exactly is a "label" ?
Thank you.
Regards Ari Kuusisto Research physicist
PerkinElmer Life and Analytical Sciences tel. +358-2-2678 508 fax. +358-2-2578 357 E-mail: Ari.Kuusisto-at-PerkinElmer.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 28 14:12:26 2004
From ESL point of view: marker is something which left "mark". So, it's tool to make "marks". Labels usually used to "label" something. So, you may "label" something to make it recognizable from something else... I would think that label is something, which someone used to mark some particular stuff. Therefore, antibody may be a label. Gold particle or fluorochrome may be a label. In general, I like second definition better. Sergey
At 06:18 AM 1/28/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Wednesday, January 28, 2004 2:05 PM To: microscopy-at-msa.microscopy.com
On Jan 28, 2004, at 6:18 AM, Kuusisto, Ari wrote:
} I have been discussing with various people the the use of the following } words } } dye } stain } label } marker } } The problem has been the use of the word "label". There exists two } opinion } about the proper use of it. The one states strictly that a label has } always } to be coupled to the target with antibody-antigen reaction. The other } school } states that a label is generally a molecule or an atom or a bead etc } that } makes the target visible in an instrument (eg. a microscope). So plese } someone, give me an explanation what exactly is a "label" ?
Dear Ari, I would define a dye as a substance that imparts color to a substrate. It need not be specific, so the color could be uniform, such as hair or cloth dyes. A stain is a substance that imparts visibility to a substrate, and in the context of microscopy a stain has some specificity. For LM a stain generally imparts color (i.e., is a type of dye), and for EM a stain is a stronger scatterer of electrons than is the substrate. A marker is a substance that adds a recognizable feature to a substrate. Finally, a label is a marker that is associated with a particular component of the substrate. Thus, there are fluorescent dyes that are lipophyllic, so these can be used as labels for membranes, there are molecules that bind very specifically, including biotin-avidin, so including but not restricted to antibody-antigen pairs. These can be bound to markers, such as green fluorescent protein or colloidal gold, and I call these labels. 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 Jan 28 15:00:10 2004
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From ESL point of view: marker is something which left "mark". So, it's tool to make "marks". Labels usually used to "label" something. So, you may "label" something to make it recognizable from something else... I would think that label is something, which someone used to mark some particular stuff. Therefore, antibody may be a label. Gold particle or fluorochrome may be a label. In general, I like second definition better. Sergey
At 06:18 AM 1/28/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Please respond off the list to me personally, and I will share a list with all respondents, who wish to be shared, within the next two weeks.
Thanks all,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson-at-wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/ Directions http://www.wcupa.edu/_admissions/sch_adm/directs.htm West Chester Boro http://www.west-chester.com/ Places to stay http://www.hotels-motels-n-more.com/PA/West-Chester.html
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 28 17:08:52 2004
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Nice definition, Bill.
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Wednesday, January 28, 2004 2:05 PM To: microscopy-at-msa.microscopy.com
On Jan 28, 2004, at 6:18 AM, Kuusisto, Ari wrote:
} I have been discussing with various people the the use of the following } words } } dye } stain } label } marker } } The problem has been the use of the word "label". There exists two } opinion } about the proper use of it. The one states strictly that a label has } always } to be coupled to the target with antibody-antigen reaction. The other } school } states that a label is generally a molecule or an atom or a bead etc } that } makes the target visible in an instrument (eg. a microscope). So plese } someone, give me an explanation what exactly is a "label" ?
Dear Ari, I would define a dye as a substance that imparts color to a substrate. It need not be specific, so the color could be uniform, such as hair or cloth dyes. A stain is a substance that imparts visibility to a substrate, and in the context of microscopy a stain has some specificity. For LM a stain generally imparts color (i.e., is a type of dye), and for EM a stain is a stronger scatterer of electrons than is the substrate. A marker is a substance that adds a recognizable feature to a substrate. Finally, a label is a marker that is associated with a particular component of the substrate. Thus, there are fluorescent dyes that are lipophyllic, so these can be used as labels for membranes, there are molecules that bind very specifically, including biotin-avidin, so including but not restricted to antibody-antigen pairs. These can be bound to markers, such as green fluorescent protein or colloidal gold, and I call these labels. 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 Jan 28 17:33:15 2004
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On Jan 28, 2004, at 6:18 AM, Kuusisto, Ari wrote:
} I have been discussing with various people the the use of the following } words } } dye } stain } label } marker } } The problem has been the use of the word "label". There exists two } opinion } about the proper use of it. The one states strictly that a label has } always } to be coupled to the target with antibody-antigen reaction. The other } school } states that a label is generally a molecule or an atom or a bead etc } that } makes the target visible in an instrument (eg. a microscope). So plese } someone, give me an explanation what exactly is a "label" ?
Dear Ari, I would define a dye as a substance that imparts color to a substrate. It need not be specific, so the color could be uniform, such as hair or cloth dyes. A stain is a substance that imparts visibility to a substrate, and in the context of microscopy a stain has some specificity. For LM a stain generally imparts color (i.e., is a type of dye), and for EM a stain is a stronger scatterer of electrons than is the substrate. A marker is a substance that adds a recognizable feature to a substrate. Finally, a label is a marker that is associated with a particular component of the substrate. Thus, there are fluorescent dyes that are lipophyllic, so these can be used as labels for membranes, there are molecules that bind very specifically, including biotin-avidin, so including but not restricted to antibody-antigen pairs. These can be bound to markers, such as green fluorescent protein or colloidal gold, and I call these labels. 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 Jan 28 17:47:00 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Microscopists,
I have been discussing with various people the the use of the following words
dye stain label marker
The problem has been the use of the word "label". There exists two opinion about the proper use of it. The one states strictly that a label has always to be coupled to the target with antibody-antigen reaction. The other school states that a label is generally a molecule or an atom or a bead etc that makes the target visible in an instrument (eg. a microscope). So plese someone, give me an explanation what exactly is a "label" ?
Thank you.
Regards Ari Kuusisto Research physicist
PerkinElmer Life and Analytical Sciences tel. +358-2-2678 508 fax. +358-2-2578 357 E-mail: Ari.Kuusisto-at-PerkinElmer.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 28 22:59:11 2004
I see that the messages look like they are being replicated by an EMail pickup service based in Europe.
I'm trying to block this, but I will be offline for the next ~ 24+ hours so I'll not be able to do anything to track success or failure until I surface.
If the block does not cure the duplicates please bear with it until I can get back on-line and investigate further.
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 29 00:50:57 2004
I would use the word label as being that which allows us to see the substance that we have probed the tissue for. In immunohisto/cytochemistry/fluorescence it would be the fluorochrome (eg FITC), enzyme (eg peroxidase) or a gold particle in immuno-EM.
For me 'marker' is a marker of disease - that is the antigen we are looking for that tells us the lineage of the cells we see etc. Med vänliga hälsningar/With best regards
Gareth
***Come to Stockholm in 2004 to the 26th IFBLS congress*** Take a look at http://www.vardforbundet.se/ifbls2004
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 Thu Jan 29 05:20:24 2004
To, MSA LIST Members, Would like to get performance feedback, from any of the list members, on the following L.Microscopy sample preparation Machines of diffrent make,for a technical review.
Here's how we, cell biologists, usually define these terms: - Dye: substance that adds color at light microscopy level. - Stain: substance that adds contrast for the EM. Has some specificity for some cellular components, but is overall non-specific as it does not differentiate between different protein species, for example. - Label: something used to localize specific cell components (proteins usually, but also lipids and carbohydrates). Usually antibodies followed by fluorescent probes or gold conjugates, but can also refer to histo-cytochemistry. - Marker: labels that are known to be specific for a given organelle/cell. For example: catalase is used as a peroxysome marker, calnexin as an ER marker, etc...
As you can see, each discipline uses its own definitions, so it would be very hard to normalize these and come up with a set of definitions that work for everyone. The important thing is to use the terminology that is generally accepted within your own discipline, so that other people understand your work and you can understand theirs!! Best
Marc
On Wednesday, January 28, 2004, at 09:18 AM, Kuusisto, Ari 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 Microscopists, } } I have been discussing with various people the the use of the following } words } } dye } stain } label } marker } } The problem has been the use of the word "label". There exists two } opinion } about the proper use of it. The one states strictly that a label has } always } to be coupled to the target with antibody-antigen reaction. The other } school } states that a label is generally a molecule or an atom or a bead etc } that } makes the target visible in an instrument (eg. a microscope). So plese } someone, give me an explanation what exactly is a "label" ? } } Thank you. } } } Regards } Ari Kuusisto } Research physicist } } PerkinElmer Life and Analytical Sciences } tel. +358-2-2678 508 } fax. +358-2-2578 357 } E-mail: Ari.Kuusisto-at-PerkinElmer.com } } } }
-- 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 Thu Jan 29 09:46:13 2004
Kathleen Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
mark.lewis-at-thermo.com wrote:
} Hello everyone ! } } I'm taking a survey to find out which type of Hematoxylin is most commonly } used in Histology and Cytology labs. } } Thanks ! } } Best regards, } } Mark } } Mark Lewis } Product Specialist } Anatomical Pathology } Clinical Diagnostics } Thermo Electron Corporation } (412) 747-4013 } (412) 788-1097 } E-mail: mark.lewis-at-thermo.com } } } } } _______________________________________________ } Histonet mailing list } Histonet-at-lists.utsouthwestern.edu } http://lists.utsouthwestern.edu/mailman/listinfo/histonet } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 29 10:52:44 2004
Free to a good home: An original Tracor Northern 5500 EDS Computer System Console (without the EDS detector). It was fully functional and working when taken out of service in 2000. You may find it useful for parts or for use if matched up to a detector. The recipient has to arrange for pickup or shipping, but we would be able to get it onto a pallet for you.
Please contact me off-list if you are interested,
Best Regards, ~Jon Dunlap
Jonathan Dunlap Analytical Laboratory Manager Osram Sylvania Products Inc. Precision Materials and Components 816 Lexington Avenue Warren, PA 16365 Ph: 814-726-6991 Fax: 814-726-6942
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 29 11:00:31 2004
} Here's how we, cell biologists, usually define these terms: } - Dye: substance that adds color at light microscopy level. } - Stain: substance that adds contrast for the EM. Has some } specificity for some cellular components, but is overall } non-specific as it does not differentiate between different } protein species, for example.
There are also the classics, such as Gram stain, etc., that are LM stains.
} - Label: something used to localize specific cell components } (proteins usually, but also lipids and carbohydrates). Usually } antibodies followed by fluorescent probes or gold conjugates, } but can also refer to histo-cytochemistry. } - Marker: labels that are known to be specific for a given } organelle/cell. For example: catalase is used as a } peroxysome marker, calnexin as an ER marker, etc...
Also colloidal gold used as a fiducial marker in tomography. } } As you can see, each discipline uses its own definitions, } so it would be very hard to normalize these and come up } with a set of definitions that work for everyone. The important } thing is to use the terminology that is generally accepted } within your own discipline, so that other people understand } your work and you can understand theirs!! } Agreed. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 29 11:02:07 2004
hello, everyone, who has Nanoscope IV Controller manual (Revision B, Software Version 5.12r3)? Can you send me a PDF format of this manual? I have a manual for Nanoscope IIIa and cannot get a surface potential imaging according to its instructions. Since the controller has been upgraded to IV, I want to have a look at the new manual to see what's the differences.
Thanks.
Minjun
Dept. of Electrical Engineering Univ. of Notre Dame 574-631-4916
----- Original Message ----- } From: "JUDE Dmello" {j_dmello-at-yahoo.com} To: "MicroscopyListserver" {microscopy-at-msa.microscopy.com} Sent: Thursday, January 29, 2004 6:31 AM
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Here's how we, cell biologists, usually define these terms: - Dye: substance that adds color at light microscopy level. - Stain: substance that adds contrast for the EM. Has some specificity for some cellular components, but is overall non-specific as it does not differentiate between different protein species, for example. - Label: something used to localize specific cell components (proteins usually, but also lipids and carbohydrates). Usually antibodies followed by fluorescent probes or gold conjugates, but can also refer to histo-cytochemistry. - Marker: labels that are known to be specific for a given organelle/cell. For example: catalase is used as a peroxysome marker, calnexin as an ER marker, etc...
As you can see, each discipline uses its own definitions, so it would be very hard to normalize these and come up with a set of definitions that work for everyone. The important thing is to use the terminology that is generally accepted within your own discipline, so that other people understand your work and you can understand theirs!! Best
Marc
On Wednesday, January 28, 2004, at 09:18 AM, Kuusisto, Ari 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 Microscopists, } } I have been discussing with various people the the use of the following } words } } dye } stain } label } marker } } The problem has been the use of the word "label". There exists two } opinion } about the proper use of it. The one states strictly that a label has } always } to be coupled to the target with antibody-antigen reaction. The other } school } states that a label is generally a molecule or an atom or a bead etc } that } makes the target visible in an instrument (eg. a microscope). So plese } someone, give me an explanation what exactly is a "label" ? } } Thank you. } } } Regards } Ari Kuusisto } Research physicist } } PerkinElmer Life and Analytical Sciences } tel. +358-2-2678 508 } fax. +358-2-2578 357 } E-mail: Ari.Kuusisto-at-PerkinElmer.com } } } }
-- 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 Thu Jan 29 15:55:16 2004
We are a pathology lab for a research group and use Hematoxylin 2 (Richard-Allan Scientific)for routine H & E's. We also use Gill's III.
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] Sent: Thursday, January 29, 2004 11:04 AM To: mark.lewis-at-thermo.com; Microscopy-at-sparc5.microscopy.com
We use Gill's.
Kathleen Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
mark.lewis-at-thermo.com wrote:
} Hello everyone ! } } I'm taking a survey to find out which type of Hematoxylin is most commonly } used in Histology and Cytology labs. } } Thanks ! } } Best regards, } } Mark } } Mark Lewis } Product Specialist } Anatomical Pathology } Clinical Diagnostics } Thermo Electron Corporation } (412) 747-4013 } (412) 788-1097 } E-mail: mark.lewis-at-thermo.com } } } } } _______________________________________________ } Histonet mailing list } Histonet-at-lists.utsouthwestern.edu } http://lists.utsouthwestern.edu/mailman/listinfo/histonet } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 29 16:13:31 2004
We are interested in software for a CCD camera on an optical microscope that we use for sample screening and documentation, and experiment design, in the course of doing TEM on microelectronic devices. We already have the Sony ProScan CCD camera on an Olympus MX-50, but we want software to drive the camera and rapidly facilitate measurement and annotation of features seen in the microscope. Any suggestions would be appreciated. Thanks.
John Mardinly Intel
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 29 18:26:06 2004
Recently, I bought C nanotube samples from Nanostructured & Amorphous Materials,Inc. However, it was not good for my experiments. If someone want to buy this sample, please contact me.
I have 5 g, and I will keep it ~0.3 g. Please look at the following site. The sample number is MWNT (95+%, OD 40-70 nm), and Stock #: 1242XH. http://www.nanoamor.com/inc/pdetail?v=1&pid=220
I can provide some TEM images upon request next week. According to their sample description, it is more than 95% quality. I do not promise this quality, but there is no contamination I made. The price was 70 dollars plus tax. My asking price is 45 dollars plus shipping cost. .
Thank you, Hiromi Konishi, Ph.D. The University of New Mexico
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 30 15:01:53 2004
I am looking for a compact pump for perfusion fixation of mouse brain, the liquid flow of the pump needs to be controlled by pressure, unfortunately the most products around are only gauged by the flow rate.
Can anyone help me out? Thanks in advance.
Xinran
Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center at Dallas Phone: 214-648-1830 Fax: 214-648-1801 E-mail: Xinran.Liu-at-utsouthwestern.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 30 18:52:43 2004
Do it with gravity. 1 meter is perfectly fine for mouse brain perfusion. You could not measure pressure in peristaltic pump commonly used for such purpose because of pulsation. If you will see liquid coming from the nose - it actually means the pressure is to high. Meantime it's vary from mouse to mouse.You also could measure the flow rate. It seems to me that 1 ml/min is optimal for perfusion. Sergey
At 01:14 PM 1/30/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
} I think the short answer will be that such materials don't diffract. } The spacings are in a whole other realm of size. You would need a } much longer wavelength to get diffraction. Also, the spacings are } not so regular as to be able to generate anything like planes of } atoms. "Peaks" would likely be poorly defined, but variation in } their shape as well as position might lend some usable information. } } Warren }
I hate to disagree but not only can you do electron diffraction in a TEM from such structures, I have done it myself. It is even possible to get a diffraction pattern from the bars of a grid.
As an alternative to electron diffraction, you can do light diffraction from the negative or do an FFT from a digital image.
In all cases, you very quickly get information on the average spacing of regular structures and avoid a lot of tedious measurements.
Regards, -- Larry Stoter NOTE - any message other than plain text will be automatically deleted :-)
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 31 14:40:09 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rburke-at-uvic.ca) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, January 30, 2004 at 13:53:06 ---------------------------------------------------------------------------
Question: I would like to obtain a fluorescence source for a Nikon TMD inverted microscope. I require a lamphouse, powersupply and parts to attach it to the microscope. I have the nosepiece and filter cube assembly. Nikon no longer supports this equipment, but there companies that make fluorescence units and adaptors. Is there anyone who has experience with this or knows of a supplier that may help?
-- Hi everybody, We have to confirm the identity of polypropylene and polyethylene in a film embedded in a resin using TEM. We cut with the ultramicrotome the sections and stain with OsO4 during 30 minutes. It is the first time that we do this and the results aren´t good, because there isn´t reaction with the OsO4. Has everyone done this type of work? We need sugerences. A lot of thanks in advance
************************************************************* Dra. María Belén López Mosquera Unidade de Microscopía Servicios Xerais de Apoio á Investigación Universidade da Coruña Edificio Servicios Centrais de Investigación Campus de Elviña s/n E-15071 A Coruña
} From 'Polymer Microscopy' by Saywer and Grubb I got the idea that it's not the energy spread of the primary beam that limits resolution but the energy spread due to the specimen. I assume that the same formula for the disk of least confusion applies no matter what the source of deltaE, but I'm wondering which energy spread is really limiting for high resolution TEM of 2D protein crystals for example.
Here comes a somewhat lengthy discussion that I wrote when I read that book. I'd be grateful if somebody could tell me whether I'm on the right track:
The most probable inelastic interaction is the 'plasmon production' with a most probable energy
loss to the scattered electrons of around 25 eV for carbon (EELS Atlas).
The mean free path for plasmon loss scattering is in the order of 100 nm for 100 keV electrons
(table in Williams and Carter, page 657, which unfortunately doesn't include carbon).
I think the argument used in 'Polymer Microscopy' by Sawyer and Grubb runs as follows:
} From the data above an electron loses about 25 eV of energy every 100 nm or 0,25 eV per nm of
specimen thickness it traverses.
Since this energy loss is stochastic the beam also suffers an energy-spread deltaE of about
the same magnitude. Due to the chromatic aberration of the objective lens this energy-spread limits the
attainable resolution to a value that depends on the thickness given by the following rule of thumb:
One should not expect to resolve details smaller than one tenth of the specimen thickness for
carbonacious specimens.
My comment: This argument is only valid for specimens that are at least 100 nm thick. If the specimen
is considerably thinner than the mean free path for inelastic scattering (for example 10-20 nm as is usual
in structure determination of proteins (excluding viruses)) only relatively few electrons have been
inelastically scattered at all and it doesn't make sense to speak of an energy spread due to the specimen.
The unscattered (which form the majority) and elastically scattered electrons have the same energy-spread
as the incoming electrons. Most of the inelastically scattered electrons have an energy spread that's given
by the width of the plasmon loss curve, that means a deltaE of about 50 eV (Ahn, Krinanek: EELS-Atlas).
In the weak phase-weak amplitude approximation only the unscattered and elastically scattered electrons
are regarded as taking part in image formation, whereas the inelastically scattered electrons lead to
background noise. Therefore the energy-spread of the inelastically scattered electrons is irrelevant.
Sawyer and Grubb treat energy-loss as a continuous process (". will cause a 100 keV electron to loose
about 0.25 eV per nanometer of thickness on average."). In fact an electron either losses the energy of a
plasmon (10 to 50 eV) or it doesn't and on average it does so every 100 nm it travels through the specimen.
Only when the specimen is thicker than the mean free path does it make sense to speak of an average
energy loss per nm thickness.
For faster electrons (for example 300 keV) the mean free paths are longer and accordingly all thickness
limits higher.
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em
'This winter I am giving courses to three students, of whom one is only moderately prepared, the other less than moderately, and the third lacks both preparation and ability. Such are the burdens ...' Carl Friedrich Gauss (1810) ______________________________________________
----- Original Message ----- } From: "Ellery Frahm" {frah0010-at-umn.edu} To: {edelmare-at-MUOHIO.EDU} ; {microscopy-at-MSA.Microscopy.com} Sent: Tuesday, March 02, 2004 4:07 PM
Folks:
Thank you for the info! I was not aware of the chromatic aberation equation. Thank you.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 3 08:20:42 2004
Hello: I would try staining with Ruthenium Tetraoxide (RuO4). Even though both polymers will stain they should stain at different rates. According to "Ruthenium Tetraoxide Staining of Polymers for Electron Microscopy" by Trent et al published in Macromolecules 1983 16, 589-598 polyethylene oxide will stain faster than isotactic polypropylene. I would stain only for between 5 min and 15 min. Both RuO4 and OsO4 are very dangerous chemicals so please be sure to observe proper safety procedures. Also you will need to use a cryo-ultramicrotome or the polypropylene phase will smear. Good luck. Steve
-- Hi everybody, We have to confirm the identity of polypropylene and polyethylene in a film embedded in a resin using TEM. We cut with the ultramicrotome the sections and stain with OsO4 during 30 minutes. It is the first time that we do this and the results aren´t good, because there isn´t reaction with the OsO4. Has everyone done this type of work? We need sugerences. A lot of thanks in advance
Stephen McCartney Research Associate Materials Research Institute 2108 Hahn Hall Va Tech Blacksburg, VA 24061 540-231-9765 - phone 540-231-8517 - FAX
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 3 08:44:22 2004
To first-order plasmons don't change the angle of the electrons, so an image with the plasmons is very close to an image with the no-loss electrons shifted in focus (the chromatic aberration term). A rough approximation is to consider the electrons which have lost energy as a background term, say B, in which case your contrast goes down by ~1/(1+B) so you lose apparent resolution because your signal-to-noise ratio is getting worse. (Noise from counting statistics; there is only one electron in the column at a time and you don't collect that many in an image.) For a thicker sample you should also reduce the amplitude/intensity of the zero- loss electrons, another reduction in your contrast. Note that it is contrast relative to noise that matters. (There are some non-linear imaging effects, and the imaging theory for electrons which have lost energy is a bit more complicated than this, but these effects is probably not relevant for proteins.)
N.B. I suspect that you are understimating the mean-free path, particularly for a low-density protein.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm2-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
On Wed, 3 Mar 2004, Philip Koeck 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 } ------------------------------------------------------------------------------- } } A related question: } } } From 'Polymer Microscopy' by Saywer and Grubb I got the idea that it's not } the energy } spread of the primary beam that limits resolution but the energy spread due } to the specimen. } I assume that the same formula for the disk of least confusion applies no } matter what the source } of deltaE, but I'm wondering which energy spread is really limiting for high } resolution TEM } of 2D protein crystals for example. } } Here comes a somewhat lengthy discussion that I wrote when I read that book. } I'd be grateful } if somebody could tell me whether I'm on the right track: } } The most probable inelastic interaction is the 'plasmon production' with a } most probable energy } } loss to the scattered electrons of around 25 eV for carbon (EELS Atlas). } } The mean free path for plasmon loss scattering is in the order of 100 nm for } 100 keV electrons } } (table in Williams and Carter, page 657, which unfortunately doesn't include } carbon). } } } } I think the argument used in 'Polymer Microscopy' by Sawyer and Grubb runs } as follows: } } } From the data above an electron loses about 25 eV of energy every 100 nm or } 0,25 eV per nm of } } specimen thickness it traverses. } } Since this energy loss is stochastic the beam also suffers an energy-spread } deltaE of about } } the same magnitude. Due to the chromatic aberration of the objective lens } this energy-spread limits the } } attainable resolution to a value that depends on the thickness given by the } following rule of thumb: } } One should not expect to resolve details smaller than one tenth of the } specimen thickness for } } carbonacious specimens. } } } } My comment: This argument is only valid for specimens that are at least 100 } nm thick. If the specimen } } is considerably thinner than the mean free path for inelastic scattering } (for example 10-20 nm as is usual } } in structure determination of proteins (excluding viruses)) only relatively } few electrons have been } } inelastically scattered at all and it doesn't make sense to speak of an } energy spread due to the specimen. } } The unscattered (which form the majority) and elastically scattered } electrons have the same energy-spread } } as the incoming electrons. Most of the inelastically scattered electrons } have an energy spread that's given } } by the width of the plasmon loss curve, that means a deltaE of about 50 eV } (Ahn, Krinanek: EELS-Atlas). } } In the weak phase-weak amplitude approximation only the unscattered and } elastically scattered electrons } } are regarded as taking part in image formation, whereas the inelastically } scattered electrons lead to } } background noise. Therefore the energy-spread of the inelastically scattered } electrons is irrelevant. } } Sawyer and Grubb treat energy-loss as a continuous process (". will cause a } 100 keV electron to loose } } about 0.25 eV per nanometer of thickness on average."). In fact an electron } either losses the energy of a } } plasmon (10 to 50 eV) or it doesn't and on average it does so every 100 nm } it travels through the specimen. } } Only when the specimen is thicker than the mean free path does it make sense } to speak of an average } } energy loss per nm thickness. } } } } For faster electrons (for example 300 keV) the mean free paths are longer } and accordingly all thickness } } limits higher. } } } } } } } } Philip Koeck } Svdertvrns Hvgskola and } Karolinska Institutet } Dept. of Bioscience at Novum } S-14157 Huddinge } Sweden } phone: +46-8-6089186 } fax: +46-8-6089290 } http://www.biosci.ki.se/em } } 'This winter I am giving courses to three students, of whom one is only } moderately prepared, } the other less than moderately, and the third lacks both preparation and } ability. } Such are the burdens ...' Carl Friedrich Gauss (1810) } ______________________________________________ } } } ----- Original Message ----- } } From: "Ellery Frahm" {frah0010-at-umn.edu} } To: {edelmare-at-MUOHIO.EDU} ; {microscopy-at-MSA.Microscopy.com} } Sent: Tuesday, March 02, 2004 4:07 PM } Subject: [Microscopy] Re: KeV vs chromatic aberation } } } } } } } } -------------------------------------------------------------------------- } ---- } } 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 Richard, } } } } We just covered this a few weeks ago in my course: } } } } The equation for the diameter of the disc of least confusion (d) for } } chromatic aberration is: } } } } d = Cc . alpha . (delta E / Eo) } } } } Cc is the coefficient for chromatic aberration, alpha is the convergence } } angle of the beam, delta E is the energy difference, and Eo is essentially } } the beam energy. For thermionic emission from a tungsten filament, the } } initial energy of the electrons varies between about 0 and 2 eV or so -- } } this is, I believe, a function of variations in their intitial thermal } } energies within the filament. } } } } As a result, if the accelerating voltage is 2 kV, (delta E / Eo) is 0.001; } } if the accelerating voltage is 20 kV, (delta E / Eo) becomes 0.0001; and } if } } the accelerating voltage is 200 kV, (delta E / Eo) is 0.00001. Thus, the } } diameter of the disc of least confusion for chromatic aberration is } } basically inversely proportional to the accelerating voltage you're using. } } } } Hope this helps, } } } } Ellery } } } } --------------- } } Ellery E. Frahm } } Research Scientist/Manager } } Electron Microprobe Laboratory } } Department of Geology & Geophysics } } University of Minnesota - Twin Cities } } Lab Website: http://probelab.geo.umn.edu } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 3 09:20:03 2004
We are looking for a new critical point dryer and would appreciate any feedback users can provide. We currently have a Polaron Jumbo, that is about 30 years old. It works, but the temperature rises very high - it is controlled just by hot water - and each run seems to have different conditions. We would like to get a machine that might be more constant from one run to the next, and that students might find somewhat easier to use. Most of the CPDs we have seen through our web hunts seem to have very small sample chambers, though. We would like to know 1) what CPDs people like, 2) whether sample chamber size seems to be a problem, 3) any other concerns that users have and would alert us to. Many thanks,
Dr. Sally Leys Assistant Professor and Canada Research Chair in Evolutionary Developmental Biology Department of Biological Sciences CW 405 The University of Alberta Edmonton, Alberta Canada T6G 2E9
How can we best prepare our soil samples for SEM and micro analysis? We are novices at this. We have great equipment (Hitachi FESEM 4500 and EDAX Genesis) We are interested in phosphorus and iron in river sediments. We are having difficulty in interpreting the results we have been getting (we crushed soil to double sticky tape, mounted on Al stubs and coated with AuPd). The preps look good at the SEM with little charging.
What we see is grains and some organic matter (OM) and coatings. The spectrum shows some OM and SiO2 along with other particles of mixed composition.
My question is how can we interpret this? What about references etc? We need your expertise!
Your help will be greatly appreciated. Carol Bronick
Agricultural Research Station Box 9061 Virginia State University Petersburg, VA 23806 804.524.6822 cbronick-at-vsu.edu
____________________________________________________________________ Check your SchoolEmail at http://www.CampusI.com Find the LOWEST PRICES on books at http://www.campusi.com/BookFind !
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 3 09:57:51 2004
Slight correction to my previous post. Of course you will have problems with both the polyethylene and polypropylene if you don't use a cryo system when microtoming. Steve
Hello: I would try staining with Ruthenium Tetraoxide (RuO4). Even though both polymers will stain they should stain at different rates. According to "Ruthenium Tetraoxide Staining of Polymers for Electron Microscopy" by Trent et al published in Macromolecules 1983 16, 589-598 polyethylene oxide will stain faster than isotactic polypropylene. I would stain only for between 5 min and 15 min. Both RuO4 and OsO4 are very dangerous chemicals so please be sure to observe proper safety procedures. Also you will need to use a cryo-ultramicrotome or the polypropylene phase will smear. Good luck. Steve
Stephen McCartney Research Associate Materials Research Institute 2108 Hahn Hall Va Tech Blacksburg, VA 24061 540-231-9765 - phone 540-231-8517 - FAX
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 3 10:52:52 2004
Dear Sally, I recently needed to buy a CPD for my new lab and after looking into the options I bought a Tousimis Samdri PVT-3D. This unit has cooling for the chamber provided by letting CO2 from the tank run past the chamber, so it is easy to cool. I like not having to mess with water cooling. I also like to do my ethanol CO2 exchanges around 0C. The chamber size is 1.25 in round by 1.25 in tall. There is an accessory available to make a much bigger chamber (not wider, just longer). I also have been extremely impressed by the Tousimis company support, with nice instructions and extras, and prompt responses to my frequent questions.
I have no financial interest in the company, just a happy customer,
Tobias
} } We are looking for a new critical point dryer and would appreciate } any feedback users can provide. We currently have a Polaron Jumbo, } that is about 30 years old. It works, but the temperature rises very } high - it is controlled just by hot water - and each run seems to } have different conditions. We would like to get a machine that might } be more constant from one run to the next, and that students might } find somewhat easier to use. Most of the CPDs we have seen through } our web hunts seem to have very small sample chambers, though. We } would like to know 1) what CPDs people like, 2) whether sample } chamber size seems to be a problem, 3) any other concerns that users } have and would alert us to. } Many thanks, } } Dr. Sally Leys } Assistant Professor and Canada Research Chair } in Evolutionary Developmental Biology } Department of Biological Sciences CW 405 } The University of Alberta } Edmonton, Alberta } Canada } T6G 2E9 } } Telephone: (780) 492-6629 } Fax: (780) 492-9234 } Email: sleys-at-ualberta.ca
The previous responders have made mostly made good points, but have neglected one important possibility. Zeiss planapo lenses of that era are of high optical quality, but unfortunately they frequently suffer from a separation of lens elements known as delamination, caused by a failure of the lens cement. Used objectives also frequently have dust or a deposit of fine, misty oil droplets on the surface of the back lens. If you have a "phase telescope" you should focus through the objective and look for contamination, or delamination, which may appear as gray patches or show interference colors, and usually start at the edges and work their way inward. You can also use a dissecting microscope to look through the objective (put a light source in front of the objective and look through the back).
Either contamination or delamination will cause a haziness of the image, which will be reduced with stopping down of the condenser, explaining why the best image is seen when the condenser is stopped down past it's theoretical optimal position. Stopping down also causes dirt or other artifacts in the optical path to become much more noticeable. If you can't locate the points of contamination with the phase telescope, try rotating the objective to see if the artifacts move with it. Haze, dirt, and delamination may also be present in the condenser. Most used objectives and condensers I have come across were in need of a good cleaning.
If you want to use your 100x planapo or any other oil lens, not using oil between the specimen and the objective is not an option. With my Zeiss microscope and 1.4 NA condenser, oiling the condenser noticeable improves the contrast as well as resolution. As others have said, you should definitely not oil your 0.9 NA condenser. Also be aware that different types of immersion oil should never be allowed to mix, and that some types of oil can damage the mounting cement of some brands of lenses. Using Zeiss immersion oil with some Nikon lenses, for example, can be disastrous.
Ralph Common Electron Microscopist Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824 517-355-7558; fax 517-432-1053 ralph.common-at-ht.msu.edu
-----Original Message----- } From: Bruce Girrell [mailto:bigirrell-at-microlinetc.com] Sent: Thursday, February 26, 2004 5:08 PM To: Microscopy-at-msa.microscopy.com
I have a binocular Zeiss phase contrast microscope that came with a basic assortment of lenses. A while back I bought a planapo 10/0.32 lens (standard - no phase ring) and was stunned at the quality of the image. To describe the image as "sharp" or even "crisp" doesn't do it justice. Some stained thin sections have the appearance of stained glass and the image almost takes on a three dimensional quality. I was impressed.
Recently, I was able to acquire a planapo 100/1.3 Ph3. Despite its very impressive appearance, I have been much less impressed with the image produced by this lens. Granted, the contrast is better than the standard lens, but based on the difference I had seen at 10x I was expecting a little more in the sharpness department.
Bear in mind that I fall in the "hobbyist" ranks. Some of these questions are basic. I have set up Kohler illumination. I believe that I have the phase rings aligned. Most of these questions apply to brightfield observation anyway.
Questions: 1) Am I simply expecting too much at 1000x? 2) Is the phase ring degrading the performance? 3) My condenser only has an n.a. of 0.9. Is this the problem? 4) Even with this condenser, should I be using oil on it as well as the objective? 5) With the phase stop in place performance is worse - lots of artifacts. That is not the case with the 40x and 10x phase objectives.
One more general question: Literature tells me that I should achieve optimum contrast/resolution with the brightfield aperture closed about 1/3 of the way. I seem to get the best results with the aperture closed just a little over half way. Am I not sensitive enough to artifacts and confusing increased contrast for increased resolution? Am I maybe using too much light?
Thanks for your help,
Bruce Girrell
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 3 13:23:26 2004
This is a question about TEM objective apertures, mostly about thickness and selection of hole size.
We have a JEOL 1200, I wanted to replace the objective aperture, old one was not cleaning up well. Ordered one from EM supplier we get most everything else from. Picked one that was for JEOL, a Pt strip .1 mm thick, three holes, 50, 100, 150. Sounded perfect.
Tried to put it in yesterday. Way too thick. Aperture holder accepts the strip by slipping it in a fixed sandwich kind of space, not with a separate screw down holder over the top. Old aperture was way thinner than the new one, even though new one was called 'thin'.
Checked all the other catalogs I have, most had apertures with the same dimensions. Don't have much experience with this so I am hoping to be educated.
Are the apertures for this instrument something special? Where do I get an aperture to fit? Is JEOL the only source? What criteria do I use to select the hole size? etc.
Help me out on this one.
Thanks
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 Mar 3 13:38:11 2004
First of all, Au lines will overlap with P, so in this case coating with Au is not acceptable.
Then I am not sure about a goal of your research. If you need just to determine whether Fe and P are in your samples, then EDS is not the best choice of method. Depending on anticipated levels of elements the XRF, wet chemistry or even mass spectrometry could be a better choice.
If detection limit of 0.5% is OK, then I would use high intensity beam current (so that dead time is about 20-50%)to acquire spectrum from pretty big field of view, at magnification of x100 or x200, for at least 5 min. Sometimes fast mapping (again at high beam current and at magnifications when many particles are visible) can help to localize the place of interest.
Regards,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} Hello fellow Microscopists, } } How can we best prepare our soil samples for SEM and micro } analysis? We are novices at this. We have great equipment } (Hitachi FESEM 4500 and EDAX Genesis) We are interested in } phosphorus and iron in river sediments. We are having } difficulty in interpreting the results we have been getting } (we crushed soil to double sticky tape, mounted on Al stubs } and coated with AuPd). The preps look good at the SEM with } little charging. } } What we see is grains and some organic matter (OM) and } coatings. The spectrum shows some OM and SiO2 along with } other particles of mixed composition. } } My question is how can we interpret this? What about } references etc? We need your expertise! } } Your help will be greatly appreciated. } Carol Bronick } } Agricultural Research Station } Box 9061 } Virginia State University } Petersburg, VA 23806 } 804.524.6822 } cbronick-at-vsu.edu } } ____________________________________________________________________ } Check your SchoolEmail at http://www.CampusI.com } Find the LOWEST PRICES on books at http://www.campusi.com/BookFind ! } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 3 14:05:24 2004
Why don't you use JEOL spares as your first choice?
At least you then know that they will fit and work.
Down here, at least, JEOL parts are not exhorbitantly expensive.
cheers
rtch (a satisfied customer of JEOL Sydney)
Date sent: Wed, 3 Mar 2004 11:17:01 -0800 To: Microscopy-at-sparc5.microscopy.com } From: jmkrupp-at-cats.ucsc.edu (Jon Krupp)
Jonathan I do believe, the apertures in JEM1200 are made from molybdenum (or tungsten?), definitely not a Pt. Perhaps you have to contact JEOL for specification (they are quite friendly). Personally, I would avoid "after market" spare parts especially objective apertures (they are designed in the way they are actually self-cleaning under the beam). Aperture in my microscope is "in" for more than 5 years and I never cleaned it. Deposits on the objective aperture usually indicates problem with vacuum. You may easily clean original JEOL aperture by heating up in tantalum or tungsten boat in good vacuum (5x10-6 torr or better) for a minute or less. It cleans everything. The problem there is that it's possible to damage the hole itself (well, it means, it's time to buy new one) if overheat. It's important, the boat made from different material than aperture, otherwise, the strip will stick to the boat. I am using 25-50-100 um objective aperture strip. In 95% cases we are using 50um spot - it provides enough light with decent contrast. I am using 25um spot (not recommended to use with plastic sections!) for very weak W shadowed samples only . I never used 100 um. I hope it helps. Sergey
At 11:17 AM 3/3/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Actually, P K alpha and Au M alpha don't overlap but are indeed very close by 108eV. Pt might be an option but its M alpha is even closer to the P K alpha. The 108eV distance at low eV is not always a problem with Genesis. The EDAX HPD feature is very powerful for deconvoluting peaks. A quant with low intensity errors will give very good results. Any element that has high (} 20%) intensity error should be discarded or re-run at different KV.
At F, my detector is about 56-59eV (give or take) resolution and how one calculates this. Calibrated rez at Al and Cu is typically 128eV at 102uS. No spec wars please.
Potting the soil in metallurgical mounting media then polishing should provide good results. The irregularity of un-polished (3-D) soil grains is not good for EDS. Flat, polished surfaces are best.
Collect at 102uS with at least 1,000cps and do it for about 300 live seconds. Keep DT { 25%. Then do peak ID, HPD and finally, quant and look at the Inten. Error values.
I look at C, F, O, P, Al, Si, Fe, S, Cu, W, Ta, Hf, Zr, and many others (not all at the same time!!) that are Au/Pd coated. At 2X KV of highest value, I can't recall a time that I could not define the constituents with EDAX EDS. Organics/polymers are another story-- need FTIR or WDS for that.
gary g.
At 11:51 AM 3/3/2004, Dusevich, Vladimir wrote:
} First of all, Au lines will overlap with P, so in this } case coating with Au is not acceptable. } } Then I am not sure about a goal of your research. } If you need just to determine whether Fe and P are in } your samples, then EDS is not the best choice of method. } Depending on anticipated levels of elements the XRF, } wet chemistry or even mass spectrometry could be a better } choice. } } If detection limit of 0.5% is OK, then I would use } high intensity beam current (so that dead time is about } 20-50%)to acquire spectrum from pretty big field of view, } at magnification of x100 or x200, for at least 5 min. } Sometimes fast mapping (again at high beam current and at } magnifications when many particles are visible) can } help to localize the place of interest. } } Regards, } } Vladimir } } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } } Hello fellow Microscopists, } } } } How can we best prepare our soil samples for SEM and micro } } analysis? We are novices at this. We have great equipment } } (Hitachi FESEM 4500 and EDAX Genesis) We are interested in } } phosphorus and iron in river sediments. We are having } } difficulty in interpreting the results we have been getting } } (we crushed soil to double sticky tape, mounted on Al stubs } } and coated with AuPd). The preps look good at the SEM with } } little charging. } } } } What we see is grains and some organic matter (OM) and } } coatings. The spectrum shows some OM and SiO2 along with } } other particles of mixed composition. } } } } My question is how can we interpret this? What about } } references etc? We need your expertise! } } } } Your help will be greatly appreciated. } } Carol Bronick } } } } Agricultural Research Station } } Box 9061 } } Virginia State University } } Petersburg, VA 23806 } } 804.524.6822 } } cbronick-at-vsu.edu } } } } ____________________________________________________________________ } } Check your SchoolEmail at http://www.CampusI.com } } Find the LOWEST PRICES on books at http://www.campusi.com/BookFind ! } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 3 15:39:14 2004
In response to Sergey and Jonathan's questions let me say this: Ladd, as most people know, produces probably 97% of all apertures made in the U.S. We customize apertures for all microscopes to fit the user's needs. For instance in Sergey's case, he could have two or even three 50um holes if he wanted. The "after market" apertures offer you this customization opportunity. We can machine in various materials, such as moly, platinum, etc. and provide strips or discs as thin as 0.025mm.
Generally, apertures in the "after market" are quite a bit less expensive, more readily available and, as I mentioned, can be customized to meet your needs. We put as many as 20 holes in some plates and strips for specialized equipment.
As a matter of fact most apertures are "after market" because EM manufacturers tend not to produce their own apertures. I am not impartial on this matter, but for flexibility and value the "after market" is the place to look for apertures.
Disclaimer: Ladd Research sells EM supplies including apertures and specialized micro-holes to EM users and microscope manufacturers.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 03, 2004 3:56 PM
John I LOVE "after market" especially if it provides CHEAP solution. I also like your point on customization (which, perhaps, will be more expensive...). From another hand Jonathan's message shows the reality: he bought "after market" aperture (I would think he did specify, which aperture he needs), which does not fit... As I told in my original message, aperture on JEM1200 survived for many years and it was cost to me something like $100 from JEOL. How much I could save on "after market"? $30 perhaps or even less? To me it would be probably easier to get stuff from guaranteed source (JEOL) rather than spent time looking for aperture, which will "fit"... Sergey
At 01:52 PM 3/3/2004, you wrote: } In response to Sergey and Jonathan's questions let me say this: Ladd, as } most people know, produces probably 97% of all apertures made in the U.S. } We customize apertures for all microscopes to fit the user's needs. For } instance in Sergey's case, he could have two or even three 50um holes if he } wanted. The "after market" apertures offer you this customization } opportunity. We can machine in various materials, such as moly, platinum, } etc. and provide strips or discs as thin as 0.025mm. } } Generally, apertures in the "after market" are quite a bit less expensive, } more readily available and, as I mentioned, can be customized to meet your } needs. We put as many as 20 holes in some plates and strips for specialized } equipment. } } As a matter of fact most apertures are "after market" because EM } manufacturers tend not to produce their own apertures. I am not impartial } on this matter, but for flexibility and value the "after market" is the } place to look for apertures. } } Disclaimer: Ladd Research sells EM supplies including apertures and } specialized micro-holes to EM users and microscope manufacturers. } } John Arnott } } Ladd Research } 83 Holly Court } Williston, VT 05495 } } On-line Catalog: http://www.laddresearch.com } } tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) } fax: 1-802-660-8859 } e-mail: sales-at-laddresearch.com } ----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, March 03, 2004 3:56 PM } Subject: [Microscopy] Re: TEM apertures } } } } } } } } -------------------------------------------------------------------------- } ---- } } 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 } } -------------------------------------------------------------------------- } ----- } } } } Jonathan } } I do believe, the apertures in JEM1200 are made from molybdenum (or } } tungsten?), definitely not a Pt. Perhaps you have to contact JEOL for } } specification (they are quite friendly). Personally, I would avoid "after } } market" spare parts especially objective apertures (they are designed in } } the way they are actually self-cleaning under the beam). Aperture in my } } microscope is "in" for more than 5 years and I never cleaned it. Deposits } } on the objective aperture usually indicates problem with vacuum. You may } } easily clean original JEOL aperture by heating up in tantalum or tungsten } } boat in good vacuum (5x10-6 torr or better) for a minute or less. It } } cleans everything. The problem there is that it's possible to damage the } } hole itself (well, it means, it's time to buy new one) if overheat. It's } } important, the boat made from different material than aperture, otherwise, } } the strip will stick to the boat. I am using 25-50-100 um objective } } aperture strip. In 95% cases we are using 50um spot - it provides enough } } light with decent contrast. I am using 25um spot (not recommended to use } } with plastic sections!) for very weak W shadowed samples only . I never } } used 100 um. I hope it helps. Sergey } } } } At 11:17 AM 3/3/2004, you 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 } } } } --------------------------------------------------------------------------- } ---- } } } } } } Greetings: } } } } } } This is a question about TEM objective apertures, mostly about thickness } } } and selection of hole size. } } } } } } We have a JEOL 1200, I wanted to replace the objective aperture, old one } } } was not cleaning up well. Ordered one from EM supplier we get most } } } everything else from. Picked one that was for JEOL, a Pt strip .1 mm } thick, } } } three holes, 50, 100, 150. Sounded perfect. } } } } } } Tried to put it in yesterday. Way too thick. Aperture holder accepts the } } } strip by slipping it in a fixed sandwich kind of space, not with a } separate } } } screw down holder over the top. Old aperture was way thinner than the new } } } one, even though new one was called 'thin'. } } } } } } Checked all the other catalogs I have, most had apertures with the same } } } dimensions. Don't have much experience with this so I am hoping to be } } } educated. } } } } } } Are the apertures for this instrument something special? Where do I get } an } } } aperture to fit? Is JEOL the only source? What criteria do I use to } select } } } the hole size? etc. } } } } } } Help me out on this one. } } } } } } Thanks } } } } } } Jonathan Krupp } } } Microscopy & Imaging Lab } } } University of California } } } Santa Cruz, CA 95064 } } } (831) 459-2477 } } } jmkrupp-at-cats.ucsc.edu } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } 10833 Le Conte Ave, Room 33-089 } } Los Angeles, CA 90095 } } } } Phone: (310) 825-1144 (office) } } (310) 206-1029 (Lab) } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } } } } } }
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jvonreis-at-columbiabasin.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 3, 2004 at 11:20:34 ---------------------------------------------------------------------------
Email: jvonreis-at-columbiabasin.edu Name: Jennifer von Reis
Question: Does anyone have information on how to use hexamethyldisilazene (HMDS) instead of a critical point drier to prepare specimens, that have been fixed and dehydrated in alcohol, for SEM observation?
We have done quite a bit so far. Most investigations were done on on sidiments ranging between 1m below the surface up to rocks at ~100 m Below the surface. My preference is still to mount in Araldite (the cheap version you can get from fiberglass hobby shops) under vacuum to get rid of trapped gas and polish to 1 micron surface finish. Do not crush. You losse info like pore density and distibution/relationship of the different components. Weathering is more clear in a cross section. Most of the investigation is in BSE mode. Carbon coating is preferred since it interfere less with the EDS spectrum. If you can work in Low Vacuum range (0.1 torr - 1 torr) it helps with reducing charging.
A reasonable refernce is (more suitable to rocks) is "Bachscattred Scanning Electron Microscopy and Image analysis of Sediments and Sedimentary Rocks" by D. H. Kringsly and other authers. Cambridge press ISBN 0-521-45346-1
How can we best prepare our soil samples for SEM and micro analysis? We are novices at this. We have great equipment (Hitachi FESEM 4500 and EDAX Genesis) We are interested in phosphorus and iron in river sediments. We are having difficulty in interpreting the results we have been getting (we crushed soil to double sticky tape, mounted on Al stubs and coated with AuPd). The preps look good at the SEM with little charging.
What we see is grains and some organic matter (OM) and coatings. The spectrum shows some OM and SiO2 along with other particles of mixed composition.
My question is how can we interpret this? What about references etc? We need your expertise!
Your help will be greatly appreciated. Carol Bronick
Agricultural Research Station Box 9061 Virginia State University Petersburg, VA 23806 804.524.6822 cbronick-at-vsu.edu
____________________________________________________________________ Check your SchoolEmail at http://www.CampusI.com Find the LOWEST PRICES on books at http://www.campusi.com/BookFind !
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 4 00:25:17 2004
-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Jennifer von Reis wrote: ================================================================== Question: Does anyone have information on how to use hexamethyldisilazene (HMDS) instead of a critical point drier to prepare specimens, that have been fixed and dehydrated in alcohol, for SEM observation? ================================================================== A good overview on this topic was published in MICROSCOPY TODAY, May 1997 by Phil Oshel. It has been republished on the SPI Supplies website with permission at URL http://www.2spi.com/catalog/chem/hmds.html
Disclaimer: SPI Supplies is a supplier of HMDS used in electron microscopy applications.
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 Mar 4 00:49:29 2004
Sergey; Best way to clean apertures is with a dual beam FIB. That way, you can cut away any dirt, burs, eccentricity, and inspect with the SEM while you are in there!
John Mardinly Intel
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Wednesday, March 03, 2004 12:57 PM To: Microscopy-at-sparc5.microscopy.com
Jonathan I do believe, the apertures in JEM1200 are made from molybdenum (or tungsten?), definitely not a Pt. Perhaps you have to contact JEOL for specification (they are quite friendly). Personally, I would avoid "after market" spare parts especially objective apertures (they are designed in
the way they are actually self-cleaning under the beam). Aperture in my
microscope is "in" for more than 5 years and I never cleaned it. Deposits on the objective aperture usually indicates problem with vacuum. You may easily clean original JEOL aperture by heating up in tantalum or tungsten boat in good vacuum (5x10-6 torr or better) for a minute or less. It cleans everything. The problem there is that it's possible to damage the hole itself (well, it means, it's time to buy new one) if overheat. It's important, the boat made from different material than aperture, otherwise, the strip will stick to the boat. I am using 25-50-100 um objective aperture strip. In 95% cases we are using 50um spot - it provides enough
light with decent contrast. I am using 25um spot (not recommended to use with plastic sections!) for very weak W shadowed samples only . I never
used 100 um. I hope it helps. Sergey
At 11:17 AM 3/3/2004, you wrote:
} ----------------------------------------------------------------------- ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
A lot of thanks for the information! I am sure that I will contact with you. Thank you -- ************************************************************* Dra. María Belén López Mosquera Unidade de Microscopía Servicios Xerais de Apoio á Investigación Universidade da Coruña Edificio Servicios Centrais de Investigación Campus de Elviña s/n E-15071 A Coruña
O.k., folks I know a number of you are still using TEM film as we are. And due to the pricing of the 250 sheet boxes vs. the 100 sheet boxes you're buying the 250 sheet boxes. Great deal on film cost but the lousy plastic boxes aren't much use.
Those little yellow kodak (or white Ilford) 100 sheet film boxes are great for lots of things but we've been using them for transporting exposed film from the scope rooms to the dark room for processing. I assume that other folks have been doing the same since I've come across this practice at every TEM lab I've ever worked at. But since we've been buying the plastic 250 sheet boxes our supply of the yellow cardboard boxes has been dwindling slowly. SO now the quesiton is what do we replace the 100 sheet cardboard boxes with? Has anyone found the answer yet?
The $40 price premium for buying 100 sheet boxes of film is a bit much for replacing the cardboard boxes withy new cardboard boxes.
The $20K and upwards price for a CCD camera to move to digital is also a bit much.
So before I have my shop folks here make me up some replacment metal boxes, are there any other suggestions out there?
Any of the TEM FIlm vendors out there willing to sell expired 100 sheet film really cheap? (You can even keep the film!)
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 4 08:15:31 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (R.H.Olley-at-reading.ac.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, March 4, 2004 at 03:53:49 ---------------------------------------------------------------------------
Email: R.H.Olley-at-reading.ac.uk Name: Robert H. Olley
Title-Subject: [Microscopy] [Filtered] TEM of Polymers
Question: In answer to the question from Dra. Maria BelÈn LÛpez Mosquera:
One way of looking at polymers, especially hydrocarbon polymers like polypropylene and polyethylene, is to prepare a surface and etch with a permanganic reagent. The etched surface is then either: - replicated and the replica examined under TEM; - gold coated and examined under SEM.
It is very easy to distinguish polyethylene and polypropylene this way.
We have a picture gallery of etched surfaces, albeit for specimens with special thermal or mechanical treatment, on:
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.personal.rdg.ac.uk/~spsolley -----------------------------------